JP2020029428A - Yamabushitake-derived novel compound - Google Patents

Abstract

To provide a novel compound obtained from Yamabushitake (Hericium erinaceum) extract.SOLUTION: As one embodiment of the present invention, a compound represented by the following structure or a salt thereof, and a plant growth regulator containing the compound as an active ingredient are provided.SELECTED DRAWING: None

Description

  The present invention relates to a novel compound, and more specifically, to a compound contained in an extract of Helicium erinaceum and having a plant growth regulating activity. The present invention also relates to a plant growth regulator containing the above compound as an active ingredient.

  Yamabushitake (Hericium erinaceum) is an edible mushroom belonging to the genus Coralum agaricaceae of the order Corylus agaricus. The group of the present inventors has reported that the fruiting body and mycelium contain substances expected to be useful in the prevention and treatment of dementia (Non-Patent Documents 1 to 6). Report that substances containing cytotoxicity to HeLa cells are contained (Non-Patent Documents 7 and 8) and the like.

Tetrahedron Lett., 32, 4561-4564 (1991) Phytochemistry, 32, 175-178 (1993) Tetrahedron Lett., 35, 1569-1572 (1994) Heterocycl. Commun., 2, 51-54 (1996) Tetrahedron Lett., 37, 7399-7402 (1996) Biosci. Biotechnol. Biochem., 64, 2402-2405 (2000) Chem. Lett., 21, 2475-2476 (1992) Phytochemistry, 34, 1445-1446 (1993)

Higher fungi that form mushrooms have a life cycle of spores from hyphae, hyphae from fruiting bodies, and fruiting bodies from spores, and it is thought that various substances are produced at each stage. The full picture is not yet clear.
It is generally known that a liquid is secreted on the mycelium surface when a mushroom is generated, but this liquid has not received much attention so far.

The present inventors first speculated that the above-mentioned liquid secreted at the time of mushroom development might be deeply involved in fruiting body formation, and named this liquid Fluting Liquid (FL). Its biological activity was studied. As a result, it has been found that Yamabushitake FL exhibits a fruiting body formation-inducing activity or a fruiting body formation promoting activity against Enokitake mushrooms.
Based on these findings, the present inventors further studied the isolation of a novel compound having a biological activity mainly related to mushroom growth from each stage of the life cycle and FL of Yamabushitake, and studied several useful compounds. And found a useful biological activity, specifically, an activity of regulating plant growth.
That is, the present invention provides the following.

２． 下記の構造で示される(2S,3S,4S)-2,3,4,5-テトラヒドロキシペンチル 4-ヒドロキシ-3-(3-メチルブタノイル)ベンゾエート又はその塩。

３． 下記の構造で示される(2S,3S,4S)-2,3,4,5-テトラヒドロキシペンチル 4-クロロ-3,5-ジメトキシベンゾエート又はその塩。

４． 下記の構造で示される2,3,4,5-テトラヒドロキシペンチル 4-クロロ-3,5-ジメトキシベンゾエート又はその塩。

５． 下記の化合物又はその塩のいずれか１種以上を有効成分として含有する、植物成長調節剤。
(i) (2,2-ジメチル-2H-クロメン-6-イル)メタノール

(ii) 1-(2-ヒドロキシ-5-(ヒドロキシメチル)フェニル)-3-メチルブタン-1-オン

(iii) (4-クロロ-3,5-ジメトキシフェニル)メタノール

(iv) (2S,3S,4S)-2,3,4,5-テトラヒドロキシペンチル 4-ヒドロキシ-3-(3-メチルブタノイル)ベンゾエート

(v) (2S,3S,4S)-2,3,4,5-テトラヒドロキシペンチル 4-クロロ-3,5-ジメトキシベンゾエート

(vi) 2,3,4,5-テトラヒドロキシペンチル 4-クロロ-3,5-ジメトキシベンゾエート

1. 1- (2-hydroxy-5- (hydroxymethyl) phenyl) -3-methylbutan-1-one represented by the following structure or a salt thereof.

2. (2S, 3S, 4S) -2,3,4,5-tetrahydroxypentyl 4-hydroxy-3- (3-methylbutanoyl) benzoate represented by the following structure or a salt thereof.

3. (2S, 3S, 4S) -2,3,4,5-tetrahydroxypentyl 4-chloro-3,5-dimethoxybenzoate represented by the following structure or a salt thereof.

4. 2,3,4,5-tetrahydroxypentyl 4-chloro-3,5-dimethoxybenzoate represented by the following structure or a salt thereof.

5. A plant growth regulator comprising one or more of the following compounds or salts thereof as an active ingredient.
(i) (2,2-dimethyl-2H-chromen-6-yl) methanol

(ii) 1- (2-hydroxy-5- (hydroxymethyl) phenyl) -3-methylbutan-1-one

(iii) (4-chloro-3,5-dimethoxyphenyl) methanol

(iv) (2S, 3S, 4S) -2,3,4,5-tetrahydroxypentyl 4-hydroxy-3- (3-methylbutanoyl) benzoate

(v) (2S, 3S, 4S) -2,3,4,5-tetrahydroxypentyl 4-chloro-3,5-dimethoxybenzoate

(vi) 2,3,4,5-tetrahydroxypentyl 4-chloro-3,5-dimethoxybenzoate

  The present invention provides a novel compound derived from a natural product and having a plant growth regulating effect.

1 shows an example of a fractionation scheme for isolating compounds 1 to 3 from a culture filtrate of Yamabushitake. 1 shows an example of a fractionation scheme for isolating compounds FLY-1, FLY-4, FLY-5, FLY-7, and FLY-9 from yamabushitake fluting liquid. 1 shows a proton NMR spectrum of compound FLY-1. 1 shows a proton NMR spectrum of compound FLY-4. 1 shows a proton NMR spectrum of compound FLY-5. 1 shows a proton NMR spectrum of compound FLY-7. 1 shows a proton NMR spectrum of compound FLY-9. 1 schematically shows a method for measuring a plant growth regulating activity of a compound of the present invention. The plant growth inhibitory activities of Compounds 1 and 2 (1, 10, 100, 1000 nmol / filter paper) are shown in comparison with 2,4-D. "Control" indicates a sample to which no compound was added. The plant growth inhibitory activity of compound 3 (1, 10, 100, 1000 nmol / filter paper) is shown in comparison with 2,4-D. "Control" indicates a sample to which no compound was added.

  The present invention provides a novel compound derived from Yamabushitake and a plant growth regulator comprising a compound derived from Yamabushitake as an active ingredient. The compound of the present invention can be obtained from a Yamabushitake extract, but not particularly limited thereto, from an extract derived from a culture solution of a Yamabushitake mycelium or an extract from a Fruiting liquid (FL) of Yamabushitake.

  In the present invention, the “plant growth regulator” is intended to mean a substance having a plant growth regulation effect, and refers to a “plant growth promoter” that promotes plant growth and a “plant growth inhibitor” that inhibits plant growth. Including both. The plant growth regulator of the present invention can contain any one or more of the compounds of the present invention derived from Yamabushitake extract.

  The Yamabushitake (Hericium erinaceum) used in the present invention can be collected, for example, in Narusawa Village, Yamanashi Prefecture, but can also be cultivated artificially.

  The extract derived from the culture solution of Yamabushitake mycelium is not particularly limited. The culture broth was filtered to remove mycelium, and the obtained culture filtrate was concentrated under reduced pressure, liquid-liquid partitioned with n-hexane and ethyl acetate, and the remaining liquid was dried and solidified with ethanol. -Various soluble fractions obtained by liquid distribution can be obtained by concentration under reduced pressure.

The following compound of the present invention can be isolated from the n-hexane soluble fraction obtained in the above step. Although not particularly limited, the compound of the present invention contained in the culture solution of Yamabushitake mycelium can be separated from the hexane-soluble fraction using a means commonly used in the art such as silica gel column chromatography, reverse phase HPLC and the like. Can be isolated.
The extract derived from the culture fluid of Yamabushitake mycelium contains the following compounds (1), (2) and (3).

  The compound of formula (1) (herein referred to as compound (1)) has the chemical name of (2,2-dimethyl-2H-chromen-6-yl) methanol and is described in Examples. The hexane-soluble fraction of Yamabushitake mycelium culture filtrate was subjected to various types of chromatography using, and was isolated in a yield of 39.7 mg from 28 L of Yamabushitake mycelium culture filtrate.

  The compound of formula (2) (herein referred to as compound (2)) has the chemical name of 1- (2-hydroxy-5- (hydroxymethyl) phenyl) -3-methylbutan-1-one By using the steps described in the examples, the hexane-soluble fraction of Yamabushitake mycelium culture filtrate was subjected to various types of chromatography, whereby it could be isolated in a yield of 19.1 mg from 28 L of Yamabushitake mycelium culture filtrate. . Compound (2) was a novel compound not described in the literature.

  The compound of formula (3) (herein described as compound (3)) has the chemical name of (4-chloro-3,5-dimethoxyphenyl) methanol and uses the steps described in the Examples, By subjecting the hexane-soluble fraction of the Yamabushitake mycelium culture filtrate to various chromatographies, it could be isolated in a yield of 2.3 mg from 28 L of the Yamabushitake mycelium culture filtrate.

  It has been clarified that all of the above compounds (1) to (3) have a plant growth regulating activity. Therefore, the present invention also provides a plant growth regulator containing any one or more of compounds (1) to (3). These compounds may be those isolated from the culture medium of Yamabushitake mycelium or those chemically synthesized. Compounds (1) to (3) may be used as a free compound or as a pharmaceutically acceptable salt such as a sodium salt, a potassium salt, and an ammonium salt.

In addition, Yamabushitake secretes liquid on the surface of the mycelium when the mushroom is formed. This liquid was named fluting liquid (FL) by the inventor of the present invention, such as river bank. The fluting liquid of Yamabushitake can be obtained, for example, in an amount of about 1 L from 6,000 bacterial beds (mixture of mycelium and medium components) in an about 800 mL pot.
Yamabushitake fluting liquid extract is not particularly limited, for example, collecting fluting liquid generated when cultivating yamabushitake on a bacterial bed in Kubo Sangyo Co., Ltd., after concentration under reduced pressure, n-hexane, ethyl acetate And the remaining liquid is dried and solid-liquid partitioned with ethanol to obtain various soluble fractions, which can be obtained by concentration under reduced pressure.

  From the ethyl acetate-soluble fraction obtained in the above step, the following compound of the present invention can be isolated. Although not particularly limited, the compound that is the active ingredient in the Yamabushitake fluting liquid extract can be obtained by using a method commonly used in the art such as silica gel column chromatography, reverse phase HPLC, and the like, for example, by using an ethyl acetate-soluble fraction. Can be isolated. The extract of Yamabushitake fluting liquid contains the following compounds (FLY-1), (FLY-4), (FLY-5), (FLY-7) and (FLY-9).

  The structural formula of the compound (FLY-1) has not been identified at present. By subjecting the ethyl acetate-soluble fraction of Yamabushitake fluting liquid to various types of chromatography using the steps described in the examples, it was possible to isolate 38.8 mg of yamabushitake fluting liquid from 5.9 L.

  The compound of the formula (FLY-4) (hereinafter referred to as compound (FLY-4)) is (2S, 3S, 4S) -2,3,4,5-tetrahydroxypentyl 4-hydroxy-3- (3-Methylbutanoyl) benzoate having the chemical name, using the steps described in the Examples, by subjecting the ethyl acetate soluble fraction of Yamabushitake fluting liquid to various chromatography, Yamabushitake fluting liquid 5.9L Could be isolated in a yield of 7.4 mg.

  The compound of the formula (FLY-5) (hereinafter referred to as compound (FLY-5)) is (2S, 3S, 4S) -2,3,4,5-tetrahydroxypentyl 4-chloro-3, Having a chemical name of 5-dimethoxybenzoate and using the steps described in the Examples, the ethyl acetate soluble fraction of Yamabushitake fluting liquid was subjected to various chromatography to obtain 5.9 L to 10.5 mg of Yamabushitake fluting liquid. It could be isolated in yield.

  The compound of the formula (FLY-7) (hereinafter referred to as compound (FLY-7)) is a compound having the chemical name 2,3,4,5-tetrahydroxypentyl 4-chloro-3,5-dimethoxybenzoate. And a stereoisomer of compound (FLY-5). By subjecting the ethyl acetate soluble fraction of yamabushitake fluting liquid to various chromatography using the steps described in the examples, it was possible to isolate 39.2 mg from 5.9 liter of yamabushitake fluting liquid.

  The structural formula of the compound (FLY-9) has not been identified at present. By subjecting the ethyl acetate-soluble fraction of Yamabushitake fluting liquid to various types of chromatography using the steps described in the examples, 24.2 mg of yamabushitake fluting liquid could be isolated from 5.9 L of Yamabushitake fluting liquid.

  Compounds (FLY-4), (FLY-5) and (FLY-7) were novel compounds not described in the literature. Further, it was revealed that the compounds (FLY-1), (FLY-4), (FLY-5), (FLY-7) and (FLY-9) each have a plant growth regulating activity. Therefore, the present invention also provides a plant growth regulator containing at least one of the compounds (FLY-1), (FLY-4), (FLY-5), (FLY-7), and (FLY-9). I will provide a. These compounds may be isolated from Yamabushitake extract or may be chemically synthesized. Compounds (FLY-1), (FLY-4), (FLY-5), (FLY-7) and (FLY-9) can be used as free compounds or as pharmaceutically acceptable salts such as sodium You may use as a salt, a potassium salt, an ammonium salt, etc.

Plant growth regulator Since the compound of the present invention has a plant growth regulator activity, it can be used as a plant growth regulator. For example, a compound having a plant growth inhibitory effect can be used as an active ingredient of a plant growth inhibitor such as a herbicide. Alternatively, a compound having a plant growth promoting effect can be used alone or as an active ingredient of a plant growth promoting agent that can be added to a fertilizer or the like.
Plants whose growth can be regulated by the compounds of the present invention include, but are not particularly limited to, for example, grasses, asteraceae, cruciferous, legumes, convolvulaceae, solanaceae, and papilionidae , Asteraceae, Malvaceae, Amaranthaceae and the like.

For example, the compounds of the present invention having a plant growth inhibitory activity include: Grass weeds such as Enokorogosa, Poa annua, Poa annua, Pampas grass; It can inhibit the undesired growth of family weeds; solanaceous weeds; dianthus weeds; communisaceae weeds; mallows; weeds and the like, especially the asteraceae weeds.
On the other hand, the compound of the present invention having a plant growth promoting activity promotes the growth of, for example, rice, wheat, barley, rye, corn, sugarcane, oat, turf, chrysanthemum, sunflower, rape, soybean, tobacco, cotton, sugar beet and the like. be able to.

  The plant growth regulating activity of the compound of the present invention can be measured using an activity measuring method known in the art. The present inventors examined the promoting or inhibiting activity of compounds on elongation of hypocotyls and roots from lettuce seedlings described in the Examples. As a result, it was demonstrated that the compound of the present invention has a growth regulating activity on hypocotyl and root elongation.

The plant growth regulator of the present invention can be used by directly spraying it on, for example, a plant where growth is not desirable, specifically a garden, a paddy field, a field, an athletic field, a golf course, a vacant lot or the like where weeds can grow. In addition, it can be directly sprayed on a garden, a paddy field, a field, or the like where plants intended to promote growth grow. Alternatively, soil and / or fertilizer can be previously treated with the plant growth regulator of the present invention. The use amount of the plant growth regulator of the present invention is not limited, but may be, for example, 0.1 to 100 g, preferably 0.5 to 50 g, per are.
The plant growth regulator of the present invention includes compounds (1), (2), (3), (FLY-1), (FLY-4), (FLY-5), (FLY-7), and (FLY-9). ) May be provided alone or in combination with other agents having pesticidal activity, or together with a pesticidally acceptable excipient, as a pesticidal composition. In this case, the compound having a plant growth inhibitory activity and the compound having a plant growth promoting activity are preferably used separately according to the purpose.

  Hereinafter, the present invention will be further described with reference to examples, but the present invention is not limited to these examples.

[Example 1 Isolation of compound from culture medium of Yamabushitake mycelium]
28 L of a culture solution of Yamabushitake mycelium (dry weight: 255 g) was separated by filtration, and the obtained culture filtrate was concentrated under reduced pressure, followed by extraction with n-hexane and ethyl acetate three times each in liquid-liquid distribution. The culture filtrate after extraction was concentrated to dryness under reduced pressure, and then subjected to solid-liquid partition with ethanol. The obtained soluble parts were sequentially concentrated under reduced pressure, and each of "hexane-soluble fraction", "ethyl acetate-soluble fraction", and "ethanol-soluble fraction" was obtained as an extract derived from the culture filtrate of Yamabushitake. (FIG. 1).

  The hexane-soluble fraction obtained above is subjected to silica gel column chromatography (silica gel 60N (Kanto Chemical Co., Ltd.), solvent: dichloromethane; 90%, 80%, 70%, 60%, 50%, 40%, 30% Eluted with dichloromethane / ethyl acetate; ethyl acetate; acetone; methanol) to obtain 26 fractions. Among the fractions soluble in ethyl acetate, Fraction 8 was subjected to reverse phase HPLC (Phenylhexyl (GL Sciences, Inc.), inner diameter 20 mm × 250 mm, 70% methanol) to give Compound (1) (39.7 mg). ).

  Fraction 8 was subjected to reverse phase HPLC (Phenylhexyl (GL Sciences), internal diameter 20 mm × 250 mm, 70% methanol) to obtain compound (2) (19.1 mg). Compound (2) was found to be a novel compound that has not been reported so far.

Then, the fraction 5 was subjected to reverse phase HPLC (Phenylhexyl (GL Science Co., Ltd.), inner diameter 20 mm × 250 mm, 70% methanol) to obtain a compound (3) (2.3 mg).

Example 2 Isolation of Compound from Yamabushitake Fluting Liquid (FL)
5.9 L of Yamabushitake fungus bed of Yamabushitake fluting liquid was concentrated under reduced pressure, and then extracted with n-hexane and ethyl acetate by liquid-liquid distribution three times each. The culture filtrate after extraction was concentrated to dryness under reduced pressure, and then subjected to solid-liquid partition with ethanol. The obtained soluble parts were sequentially concentrated under reduced pressure to obtain “hexane-soluble fraction”, “ethyl acetate-soluble fraction”, and “ethanol-soluble fraction” (FIG. 2).

The ethyl acetate-soluble fraction obtained above is subjected to silica gel column chromatography (silica gel 60N (Kanto Chemical Co., Ltd.), solvent: dichloromethane; 90%, 85%, 80%, 75%, 70%, 60%, 50 %, 40%, 30% dichloromethane / methanol; methanol) and fractionated to obtain 15 fractions. Of the fractions soluble in ethyl acetate, fraction 2 was subjected to reverse phase HPLC (Phenyl (GL Sciences, Inc.), inner diameter 20 mm x 250 mm, 50% methanol) to obtain fraction 2-3. Was. Fraction 2-3 was subjected to reverse phase HPLC (Phenylhexyl (GL Sciences Inc.), inner diameter 20 mm x 250 mm, 10% methanol) to give compound (FLY-1) (38.8 mg) and compound (FLY-9) ( 24.2 mg) (FIG. 2). Both of these have not been determined, but they have been confirmed to be single compounds.
The fraction 4 was subjected to reverse phase HPLC (Phenylhexyl, inner diameter 20 mm × 250 mm, 60% methanol) to obtain a compound (FLY-4) (7.4 mg) as white crystals. The compound (FLY-4) was determined to have a molecular weight of 356 by HR-ESI-MS, and was found to have the following structure by NMR and X-ray crystal structure analysis.

  Finally, fraction 3 was subjected to reverse phase HPLC (Phenyl, ID 20 mm x 250 mm, 40% methanol) to give compound (FLY-5) (10.5 mg) and compound (FLY-7) (39.2 mg). All were obtained as white amorphous.

  Compounds (FLY-4), (FLY-5) and (FLY-7) were new compounds that had not been reported before. Proton NMR spectra of the purified compounds (FLY-1), (FLY-4), (FLY-5), (FLY-7) and (FLY-9) are shown in FIGS. 3-1 to 3-5, respectively.

[Example 3 effect]
Using compounds (1)-(3) isolated in Example 1 and 2,4-dichlorophenoxyacetic acid (2,4-D), a known herbicide component as a control compound, is illustrated in FIG. The plant was subjected to a test for controlling plant growth activity against lettuce, an Asteraceae plant, according to the following procedure.
Step 1: Pre-culture The sample solution containing each compound was infiltrated into filter paper placed in a petri dish so that the amount of the compound per filter paper was 1, 10, 100, or 1,000 nmol, and then air-dried. On the other hand, distilled water was infiltrated into a filter paper placed in another petri dish, the purpose of lettuce was set, and the cells were cultured at 25 ° C. under dark conditions for one day.
Step 2: 1 mL of distilled water was added to a filter paper obtained by air-drying the main culture sample solution, the seedlings of the precultured lettuce were transferred, and cultured at 25 ° C in a dark place for 3 days.
Step 3: Observation / Measurement The morphological change of lettuce was observed, and the hypocotyl and root elongation were measured.

  As a result, as shown in FIGS. 5 and 6, all of the compounds (1) to (3) showed significant growth inhibitory activity against the elongation of the hypocotyl and root of lettuce.

  The compound of the present invention obtained from Yamabushitake extract has a plant growth regulating activity and can be suitably used as a plant growth regulator.

Claims (5)

1- (2-hydroxy-5- (hydroxymethyl) phenyl) -3-methylbutan-1-one represented by the following structure or a salt thereof.

(2S, 3S, 4S) -2,3,4,5-tetrahydroxypentyl 4-hydroxy-3- (3-methylbutanoyl) benzoate represented by the following structure or a salt thereof.

(2S, 3S, 4S) -2,3,4,5-tetrahydroxypentyl 4-chloro-3,5-dimethoxybenzoate represented by the following structure or a salt thereof.

2,3,4,5-tetrahydroxypentyl 4-chloro-3,5-dimethoxybenzoate represented by the following structure or a salt thereof.

A plant growth regulator comprising one or more of the following compounds or salts thereof as an active ingredient.
(i) (2,2-dimethyl-2H-chromen-6-yl) methanol

(ii) 1- (2-hydroxy-5- (hydroxymethyl) phenyl) -3-methylbutan-1-one

(iii) (4-chloro-3,5-dimethoxyphenyl) methanol

(iv) (2S, 3S, 4S) -2,3,4,5-tetrahydroxypentyl 4-hydroxy-3- (3-methylbutanoyl) benzoate

(v) (2S, 3S, 4S) -2,3,4,5-tetrahydroxypentyl 4-chloro-3,5-dimethoxybenzoate

(vi) 2,3,4,5-tetrahydroxypentyl 4-chloro-3,5-dimethoxybenzoate

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