JPH04266848A - Benzyl alcohol derivative and pge2 production suppressing agent and ngf production inducing agent containing the derivative as active component - Google Patents

Abstract

PURPOSE:To provide a new benzyl alcohol derivative having PGE2-production suppressing effect and NGF-production inducing effect and useful as a PGE2- production suppressing agent and an NGF-production inducing agent. CONSTITUTION:The benzyl alcohol derivative of formula (R is carboxyl, palmitic acid residue, stearic acid residue or linoleic acid residue), e.g. 4-(3',7'-dimethyl-5'- oxo-2',6'-octadienyl)-2-formyl-3-hydroxy-5-methoxybenzyl palmitate. The compound of formula can be produced by extracting raw or dried fruit body of Hericum erinaceum with a homogeneous system consisting of water and an organic solvent, separating the liquid from the solid by filtration, centrifugal separation, etc., evaporating the organic solvent from the extract liquid to obtain an aqueous phase, extracting the aqueous phase with a mixture of water and an organic solvent to collect the organic solvent layer, removing the organic phase and subjecting to chromatographic fractionation treatment.

Description

[Detailed description of the invention]

0001

[Industrial Application Field] The present invention relates to
aceae), Hericium
Hericium ericium, a mushroom of
The present invention relates to a benzyl alcohol derivative present in the fruiting body of P. inaceum, and a PGE2 (prostaglandin E2) production inhibitor and NGF (nerve growth factor) production inducer containing the benzyl alcohol derivative as an active ingredient.

0002

BACKGROUND OF THE INVENTION There have been several reports regarding compounds contained in mushrooms and their medicinal effects. For example, Polyporus mushroom (Polyporus), a mushroom of the family Polyporus
Versicolor) contains an ergosterol derivative, and this ergosterol derivative has been shown to have a cell-killing effect on liver cancer cells.
Tetrahedron) 39,2779-2785
(1983).

[0003] Agaricus blazei, a mushroom of the Agaricaceae family, also contains ergosterol derivatives, and it has been shown in Phytochemistry 27 that these ergosterol derivatives have a cytocidal effect on cervical cancer cells.
, 2777-2789 (1988).

[0004] Similar matters are disclosed in Japanese Patent Publication No. 48-6766, Japanese Patent Application Laid-Open No. 71702-1982, and Japanese Patent Application Laid-Open No. 58-621.
It has also been reported in Publication No. 18, etc.

[0005] Further, Yamabushitake, which is a mushroom belonging to the Arinaceae family, contains phthalide derivatives and isoindolinone derivatives, and it has been shown that these phthalide derivatives and isoindolinone derivatives have a cell-killing effect on cervical cancer cells. Hedron (Tetrahedron) 3,3
73-376 (1990).

[0006]

[Problems to be Solved by the Invention] However, there are no reports on other compounds contained in Yamabushitake and their drug effects.

[0007]

[Means for solving the problem] The present inventors, however,
In view of the above-mentioned circumstances, as a result of intensive research on other compounds contained in Yamabushitake and their medicinal effects, we found that Yamabushitake contains a new benzyl alcohol derivative with a specific chemical structure. found that it has an effect of suppressing PGE2 production and an effect of inducing NGF production.

That is, the present invention provides a benzyl alcohol derivative represented by the following formula 1, a PGE2 production inhibitor containing the benzyl alcohol derivative as an active ingredient, and NG
It concerns an F production inducer.

[0009]

[Formula 1] [However, R is a palmitic acid residue, a stearic acid residue, or a linoleic acid residue excluding the carboxyl group]

0010
The benzyl alcohol derivative represented by formula 1 can be obtained by treating the fruiting body of Yamabushitake mushroom as follows. First, fresh or dried fruiting bodies of Yamabushitake are extracted with a homogeneous system of water and an organic solvent, and the organic solvent is evaporated from the extract, which is separated into solid and liquid by filtration, centrifugation, etc., to obtain an aqueous phase. In this case, as a homogeneous system of water and organic solvent, 80
~85% methanol, ethanol, 85% acetone, etc. Extraction is usually carried out at room temperature, but may also be heated under reflux, and the extraction time is usually 1 to 72 hours. For example, 85%
Add the live fruiting body of Yamabushitake mushroom to ethanol, homogenize it, leave it for a day and night at room temperature, filter it to obtain an extract, and heat the extract at 40 to 45°C under reduced pressure to evaporate the ethanol. By this, an aqueous phase is obtained.

Next, the aqueous phase is subjected to liquid-liquid distribution extraction using a mixed system of water and an organic solvent to separate an organic solvent layer, and the organic solvent is evaporated from the organic solvent layer to obtain a dry product. In this case, examples of the organic solvent include chloroform, ethyl acetate, diethyl ether, etc., but chloroform is preferable from the viewpoint of yield. For example, add chloroform to the above aqueous phase and after shaking,
The chloroform layer separated by standing is separated, and the chloroform layer is heated at 40 to 45° C. under reduced pressure to evaporate the chloroform to obtain a dry product.

[0012] The dried product itself is effective as a PGE2 production inhibitor and an NGF production inducer, but impurities are removed from the dried product to enhance its PGE2 production suppressing effect and NGF production inducing effect. Therefore, it is preferable to subject the dried product to chromatographic fractionation, and it is more preferable to further re-fractionate the chromatographically fractionated product for isolation. In this case, as described in detail later in Examples, chromatographic fractionation can be carried out using silica gel column chromatography or thin layer chromatography using hexane, chloroform, chloroform/acetone, etc. as a developing solvent, or ODS column It can be re-fractionated using high performance liquid chromatography.

The physicochemical properties and structural analysis results of the compound isolated by the re-fractionation treatment are as follows.

A compound in which R in formula 1 is a palmitic acid residue excluding a carboxyl group (hereinafter referred to as helicenone C). (1) Molecular weight: 570 (2) Infrared absorption spectrum: 1730, 1720,
1620cm-1(3) Nuclear magnetic resonance spectrum (1H
-NMR): 0.88 (t, J=0.88), 1.2
5 (m), 1.61 (m), 1.78 (s), 1.
84(s), 2.12(s), 2.33(dd, J
=7.70, 7.32), 3.01(s), 3.40
(d, J=7.33), 3.91(s), 5.32
(s), 5.32 (t, J=7.33), 6.09
(s), 6.53 (s), 10.11 (s), 12
．． 38(s) (4) Solubility in solvents: Soluble in ethyl acetate, chloroform, hexane, slightly soluble in methanol, ethanol, insoluble in water (5) Color reaction: Folin reaction positive (6) Basic, medium Distinction between acidity and acidity: Acidic substance (7) Color and shape: White crystal (melting point 39°C)

Compounds in which R in formula 1 is a stearic acid residue excluding the carboxyl group (
Hereinafter, this will be referred to as helicenone D). (1) Molecular weight: 598 Infrared absorption spectra of (2) to (6) - Distinction between basic, neutral, and acidic: Same as for herisenone C (7) Color and shape: White crystals (melting point 42°C)

001
6] A compound in which R in formula 1 is a linoleic acid residue excluding a carboxyl group (hereinafter referred to as helicenone E). (1) Molecular weight: 594 (2) Infrared absorption spectrum: Same as helicenone C (3) Nuclear magnetic resonance spectrum (1H-NMR):
0.86 (t, J=6.96), 1.27 (m),
1.62 (m), 1.78 (s), 1.84 (s),
2.04(m), 2.12(s), 2.33(d
d, J=7.69, 7.33), 2.78(dd, J=
6.60, 5.87), 3.01(s), 3.40
(d, J=7.32), 3.91(s), 5.32
(s), 5.32 (t, J = 7.32), 5.34 (
m), 6.09(s), 6.53(s), 10.
11(s), 12.38(s) (4)-(6) Solubility in solvents ~ basic, neutral,
Acidity: Same as helicenone C (7) Color and shape: Colorless oil

From the above physicochemical properties and structural analysis results, the isolated compound is a benzyl alcohol derivative represented by formula 1, among which helicenone C is a 4-(3'
,7'-dimethyl-5'-oxo-2',6'-octadienyl)-2-formyl-3-hydroxy-5-methoxybenzyl palmitate, and helicenone D is 4-(
3',7'-dimethyl-5'-oxo-2',6'-octadienyl)-2-formyl-3-hydroxy-5-
methoxybenzyl stearate, and helicenone E is 4-(3',7'-dimethyl-5'-oxo-2',6
'-octadienyl)-2-formyl-3-hydroxy-5-methoxybenzyl linoleate.

[0018] As will be described in detail later in Examples, herisenone C to E have the effect of suppressing PGE2 production and the effect of inducing NGF production.
has the effect of inhibiting arachidonic acid release. Compounds that have the effect of suppressing PGE2 production are attracting attention for their use as anti-inflammatory agents and analgesics, and compounds that have the effect of inducing NGF production are attracting attention for their use as therapeutic agents for senile dementia.

[0019]

[Example] Extraction and isolation of helicenones C to E: 7.3K of live fruiting bodies of Yamabushitake in 6 liters of 85% ethanol
g was added thereto, homogenized, left at room temperature overnight, and then filtered to obtain an extract. Four liters of 85% ethanol was added to the residue, and extraction was performed in the same manner to obtain an extract, which was combined with the first extract. Then, the combined extracts were heated at 40 to 45° C. under reduced pressure to evaporate the ethanol, thereby obtaining an aqueous phase.

One liter of chloroform was added to the aqueous phase, and after shaking, the mixture was left to stand and the separated chloroform layer was separated. One liter of chloroform was added to the residue, and the same liquid-liquid distribution extraction process was performed to separate the chloroform layer, which was combined with the first chloroform layer. The combined chloroform layers were heated at 40 to 45° C. under reduced pressure to evaporate chloroform, and further dried in a desiccator to obtain 4.99 g of a dry solid.

The dried product was dissolved in hexane and subjected to column chromatography using Wako Gel C-200 (manufactured by Wako Pure Chemical Industries, Ltd.). At this time, as developing solvents, use 60 ml each of hexane → chloroform → chloroform/acetone (8/2) so that the polarity increases sequentially.
A total of 18 fractions of 0 ml were obtained. The third and fourth fractions were subjected to high-performance liquid chromatography using an ODS column, and then re-fractionated with methanol/water (95/5). group, and from the first group, helicenone C
52.1 mg of herisenone D, 51.4 mg of herisenone D from the second group, and 6.2 mg of herisenone E from the third group were isolated.

[0022] PGE2 production inhibitory effect of hericenones C to E: 1) Preparation of rat intraperitoneal macrophages, 2) Culture in a medium containing hericenones C to E (20 hours), 3
) Macrophages were cultured in a medium containing TPA (sampled after 4 hours) using the method of Ouchi et al. (Bio
chim. Biophys. Acta, 1013,
86-91, 1989), and then radioimmunoassay (RIA) using a two-antibody method was performed on PGE2 produced by macrophages. Then, the amount of PGE2 produced in the control group cultured without administering the compound and the amount of herisenone C, herisenone D, or herisenone E were compared by 20% each.
A t-test was performed between the amount of PGE2 produced in the group cultured in the medium administered at a concentration of 5 μg/ml. As a result, each group administered hericenone C, hericenone D, or hericenone E compared to the control group had a 0.1
PGE2 production was significantly suppressed with a risk rate of %. In addition, in the above-mentioned method of Ouchi et al., the drug is added to TPA at the time of culturing in 3).
In this example, the drug was administered at the time of culturing in 2).

[0023] NGF production inducing effect of hericenones C to E:
Furukawa et al.'s method (Biochemical and B
Iophysical Research Commu
nications 136, 57-63, 1986
), late embryonic (19 days old) rat cortical primary astroglial cells were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum in an incubator (1-
(change the medium every 3 days for 2 weeks), and when it reaches confluence, add DME containing 0.5% bovine serum albumin.
The cells were changed to M and cultured for several days. Here, herisenone C, herisenone D dissolved in dimethyl sulfoxide (DMSO)
Alternatively, herisenone E was adjusted to a predetermined concentration in DMEM containing 0.5% bovine serum albumin and administered. After 24 hours of culture, the culture solution was collected and the method of Furukawa et al. (J
our own neurochemistry,
40, 734-744, 1983), the NGF concentration was measured by the enzyme immunoassay method. A t-test was conducted between a control group cultured without administering any drug and a group cultured with administration of 15 μg/ml each of herisenone C, herisenone D, or herisenone E. As a result, each group administered was significantly tested to be effective with a risk rate of 1%.

[0024]

Effects of the Invention As is already clear, the novel benzyl alcohol derivative according to the present invention described above contains PGE2
It is effective as a production inhibitor and an NGF production inducer.

Claims (1)

[Scope of Claims] [Claim 1] A benzyl alcohol derivative represented by the following formula 1. [Formula 1] [However, R is a palmitic acid residue, a stearic acid residue, or a linoleic acid residue excluding a carboxyl group] [Claim 2
A PGE2 production inhibitor comprising the benzyl alcohol derivative according to claim 1 as an active ingredient. 3. An NGF production inducer comprising the benzyl alcohol derivative according to claim 1 as an active ingredient.