WO2006093202A1

WO2006093202A1 - Phloroacylphenone glycoside, process for production of the same, antiallergic agent and antioxidant

Info

Publication number: WO2006093202A1
Application number: PCT/JP2006/303901
Authority: WO
Inventors: Kazuhisa Yasui; Syuichi Segawa; Toshio Kurihara
Original assignee: Sapporo Breweries Limited
Priority date: 2005-03-03
Filing date: 2006-03-01
Publication date: 2006-09-08
Also published as: JP2008069072A

Abstract

A phloroacyphenone glycoside represented by the general formula (1): wherein R1 represents an isopropyl, isobutyl or sec-butyl group.

Description

Specification

Fluorosylphenone glycoside, process for producing the same, antiallergic agent and antioxidant

Technical field

[0001] The present invention relates to a novel fluorosilanphenone glycoside, a method for producing the same, an antiallergic agent and an antioxidant.

Background art

[0002] Hops are the main raw material that gives the refreshing bitterness and aroma of beer. In this hop, many secondary metabolites are secreted in the Lubrin glandular hair contained in the cones, and it has become clear that these secondary metabolites greatly contribute to the bitterness and aroma of beer. More recently, these secondary metabolites have been shown to have pharmacological effects. For example, it has been shown that humulone, one of the bitter substances of beer, is effective against osteoporosis (see Non-Patent Document 1). In addition, xanthohumol and 8-prenylnaringenin, which are hop prenylflavonoids, have been shown to have anticancer activity (see, for example, Non-Patent Documents 2 and 3).

[0003] In addition, it has been reported that an effective component as an antioxidant can be obtained by adsorbing a water-soluble fraction of hop koji onto a gel-type synthetic adsorbent (see Patent Document 1).

Patent Document 1: Japanese Patent No. 3477628

Non-patent literature l: Biosci. Biotech. Biochem., 61 (1), 158, 1997

Non-Patent Document 2: Foodand Chemical Toxicology 37, 271-285, 1999 Non-Patent Document 3: European Journal of Cell Biology 80, 580-585, 2001 Disclosure of the Invention

Problems to be solved by the invention

[0004] By the way, the mechanism of allergic reaction in humans is generally as follows. In other words, in order to eliminate foreign substances (bacteria, pollen, mites, etc .: antigens, allergens) that have entered the body, biological components (antibodies, lymphocytes, etc.) are produced against them to protect the body. It is a thing. Such a mechanism for protecting the body is generally called an immune function. By the way iS Because its immune function is sensitive to foreign substances, it may be harmful to the body and cause various diseases. This form of damaging hypersensitivity to the body due to immune function is called allergy. Allergies are categorized as immediate allergies (or i-m allergies) and delayed allergies (or type IV allergies)! RU

[0005] Among allergic reactions, the onset type has a higher incidence, and the reaction mechanism is mainly considered as follows. When allergens enter the body, immunoglobulin E (igE) antibodies are produced. This IgE antibody has a strong effector effect on mast cells and basophils. Mast cells and basophils are cells having granules containing histamine and serotonin and some pharmacologically active amines. The former is present around blood vessels and connective tissue, and the latter is present in blood. A receptor that binds IgE antibody exists on the cell membrane of these cells. This receptor has the property of binding strongly to IgE antibodies. For this reason, most of the produced IgE antibody is retained in a state bound to the receptor. When the same allergen binds to the IgE antibody in such a state again, many active substances such as histamine and serotonin described above are released along with degranulation. As a result, various allergic symptoms occur.

[0006] For example, symptoms such as redness or blistering rash (urticaria) accompanied by itching on the skin, inflammation of the nose and eyes, and nasal discharge and tear secretion increase, or trachea Symptoms (such as bronchial asthma) that can cause dyspnea and dyspnea are classified as this immediate allergic disease.

[0007] Many of the conventionally used antiallergic agents are drugs against the above-mentioned diseases caused by an immediate allergic reaction whose action point is relatively clear. Examples of such drugs include antispasmodics that relax smooth muscles, sympathomimetic drugs that suppress increased capillary permeability, and antihistamines. However, these are all symptomatic drugs, and most of them are synthetic drugs. Therefore, these drugs have problems in terms of side effects such as drowsiness and high blood pressure when taken.

[0008] Accordingly, an object of the present invention is to provide a novel substance that is sufficiently excellent in antiallergic action and that is mild to the human body and skin, with sufficiently suppressed side effects. Means for solving the problem

[0009] The present inventors first used a food ingredient such as beer as a food ingredient, and extracted components from plant tissues such as ruhop are mast cells and basophils such as histamine and serotone. We thought that if the release of granules containing pharmacologically active amines could be suppressed, it would lead to the prevention and treatment of immediate allergic symptoms such as hay fever. Next, the present inventors succeeded in isolating and purifying a specific novel compound through an extraction process using hops as raw materials in various screening processes of plant tissues. And when the utility of the compound was confirmed by an assay, it was found that it was a substance that could achieve the above object, and the present invention was completed.

[0010] The present invention provides a fluorosilanone non-glycoside represented by the following general formula (1).

[Chemical 1]

Here, in the formula (1), R ¹ represents an isopropyl group, an isobutyl group or a sec-butyl group.

[0011] The fluoracylphenone glycoside of the present invention is derived from a plant tissue, and is obtained by separating the cold extract strength of a plant tissue. Here, “cold water” means water below room temperature.

In order to separate the fluorosilphonone glycoside from the cold water extract, the cold water extract may be passed through a column of a high-speed liquid chromatograph. In addition, the adsorbed component obtained by adsorbing to the synthetic adsorbent from the cold water extract of the plant tissue is eluted in methanol or a mixed solvent of ethanol and water, and the mixed solvent is passed through the above-mentioned high performance liquid chromatograph column. Liquid is preferable. In this way, the fluorosilphenone glycoside of the present invention can be more reliably separated.

[0013] In the present invention, a hop tissue may be mentioned as a plant tissue. Use hops as raw material Therefore, the fluorosilphenone glycoside of the present invention can be obtained more efficiently and reliably. The hop tissue used for extraction may be hop stems, spikelets and Z or leaves. Further, a residue obtained by extracting a vent hop or carbonic acid gas (supercritical extraction) obtained at the time of processing the concentrated hop pellet can also be used.

[0014] The inventors of the present invention conducted an in vitro study using a human basophil cell line (KU812) to inhibit the antihistamine action of the above-mentioned fluorosilan phenone glycoside by 50%. It was confirmed that the concentration (IC50) was 290 g / mL. This suggests that the fluorosylphenone glycoside of the present invention has an antiallergic action. Moreover, this fluoroacylphenone glycoside is expected to exhibit an antioxidant action and an inhibitory action on tyrosinase activity from its structure. From the above, it is useful to contain this fluorosilphenone glycoside in foods, drinks, cosmetics or pharmaceuticals.

The invention's effect

[0015] According to the present invention, it is possible to provide a substance that is sufficiently excellent in anti-allergic action and relaxed to the human body and skin with sufficient strength and side effects sufficiently suppressed.

Brief Description of Drawings

FIG. 1 is an HPLC chromatogram of polyphenol containing the fluorosilanone non-glycoside of the present invention.

FIG. 2 is a ¹ H-NMR ^ vector of the fluorosilanone glycoside of the present invention.

FIG. 3 is a ¹ H-NMR ^ vector of the fluorosilanone glycoside of the present invention.

FIG. 4 is a ¹³ C-NMR spectrum of the fluorosilanone glycoside of the present invention.

FIG. 5 is a ¹ H—H—COZY spectrum of the fluorosilanone glycoside of the present invention.

FIG. 6 is a mass spectrum of the fluorosilanone glycoside of the present invention.

FIG. 7 is a mass spectrum of sugars and known sugars obtained from the floracylphenone glycosides of the present invention.

FIG. 8 is a UV absorption spectrum of the fluorosilanone glycoside of the present invention.

FIG. 9 is a ¹ H-NMR ^ vector of the fluorosilanone glycoside of the present invention.

FIG. 10 is a ¹³ C-NMR spectrum of the fluorosilanone glycoside of the present invention.

FIG. 11 is a ¹ H— ¹ H—COZY spectrum of the fluorosilanone glycoside of the present invention. FIG. 12 is a ¹ H-NMR vector of a fluorosilanone glycoside of the present invention.

FIG. 13 is a ¹ H—H—COZY spectrum of the fluorosilanone glycoside of the present invention.

FIG. 14 is a mass spectrum of a fluorosilanone glycoside of the present invention.

FIG. 15 is a mass spectrum of the fluorosilanone non-glycoside of the present invention.

BEST MODE FOR CARRYING OUT THE INVENTION

The fluorosilphenone glycoside of the present invention is a novel compound represented by the above general formula (1) and has a glycoside structure. In the formula (1), R ¹ represents an isopropyl group, an isobutyl group or a se c butyl group. That is, the novel fluorosylphenone glycoside of the present invention comprises three types of compounds represented by the following formula (2), (3) or (4) (each represented by fluoroisobutyrophenone 2-0-β- D-Gnolecopyranoside, 1-Methylolebutyrophenone 2 -—- β-D Darcopyranoside, 2-Iso-valerophenone 2 -—- β-D Darcopyranoside).

[Chemical 2]

[0018] The florosylphenone glycoside of the present invention is derived from a plant and obtained by separation from a cold water extract of a plant tissue.

[0019] As a plant tissue, a hop cocoon flower containing a large amount of glandular tissue such as a rubulin gland, which is preferable to a tissue such as a hop stem, cocoon flower and leaf containing a relatively large amount of the above-mentioned fluorosilenone glycoside. It is more preferable that it is the structure of this part. In addition, soot hops obtained at the time of processing into concentrated hop pellets and straw after hop extract extraction by carbon dioxide extraction are also useful.

[0020] From the viewpoint of effective utilization of beer brewing by-products, hops are brewing hops. preferable.

[0021] A cold water extract of plant tissue can be obtained by a conventional method. For example, when a hop tissue is used as the plant tissue, pellets that have the tissue strength and distilled water are placed in a container and allowed to stand for a predetermined time with proper stirring. The liquid thus obtained after standing can be used as it is as a cold water extract of plant tissue. Alternatively, for example, a supernatant obtained after further centrifuging the liquid obtained after standing (hereinafter referred to as “centrifugal supernatant”) can be used as a cold water extract of plant tissue. It is. Furthermore, the liquid obtained after standing or the supernatant obtained by removing water from the centrifugal supernatant can also be used as a cold water extract.

[0022] The temperature of water when obtaining the cold water extract of the plant tissue may be a temperature that is not more than room temperature and does not freeze the water. Specifically, the temperature is preferably 0 to 50 ° C, more preferably 5 to 50 ° C, and further preferably 5 to 30 ° C. If the temperature of the water is lower than this temperature range, the extraction efficiency tends to decrease because the water freezes. When the temperature of the water is higher than this temperature range, components other than the fluorosilphenone glycosides are eluted, and therefore the treatment for separating the fluorosilphenone glycosides tends to be complicated.

Next, a preferred embodiment of the method for separating a fluorosilanone non-glycoside of the present invention will be described. The method for separating fluorosilanone glycosides of this embodiment is a step of contacting a cold extract of plant tissue with hexane to obtain a first extract on the water side (hereinafter referred to as “first step”). ), Contacting the first extract with ethyl acetate to obtain a second extract on the water side (hereinafter referred to as “second step”), contacting the second extract with butanol, A step of obtaining a third extract in the butanol (hereinafter referred to as “third step”), a step of passing the third extract through a column packed with a synthetic adsorbent and adsorbing it onto the synthetic adsorbent (hereinafter referred to as “the third adsorbent”). , Referred to as “fourth step”), a step of eluting the components adsorbed on the synthetic adsorbent into a mixed solvent of water and methanol (hereinafter referred to as “fifth step”), and a mixture containing the eluted adsorbed components. A column packed with a predetermined adsorbent It was passed through to the column of high performance liquid Kuromatodara off to obtain, comprising the step of isolating the desired fluorosilicone § Sylph enone glycoside (hereinafter referred to as "sixth step".).

[0024] (First step) In the first step, a cold water extract of plant tissue such as hops is contacted with hexane. Thereby, components of plant tissues other than the fluorosilphenone glycoside of the present invention can be extracted into hexane and selectively removed from the cold water extract. Therefore, it is possible to obtain a first extract in which the content ratio of the fluorosilphenone glycoside is further increased from the initial level. As a method for contacting the cold extract of plant tissue with hexane, for example, the above-mentioned centrifugal supernatant and hexane are placed in a separatory funnel, and then the separatory funnel is shaken to remove the cold water extract. A method of contacting with xanthan can be mentioned. When the separating funnel is allowed to stand after contacting the cold water extract with hexane, the liquid in the separating funnel is separated into water and hexane. The fluorosilphenone glycoside of the present invention is contained in water. Become.

[0025] The water collected from the separatory funnel is used as the first extract as it is as the first extract. Use it.

[0026] (Second step)

In the second step, the first extract obtained in the first step is contacted with ethyl acetate. As a result, plant tissue components other than the fluorosilacinone glycoside of the present invention can be extracted into ethyl acetate and further selectively removed from the first extract. Therefore, it is possible to obtain a second extract in which the content ratio of phenacylphenone glycoside is further increased than that of the first extract. Examples of the method of contacting the first extract with ethyl acetate include, for example, putting the first extract and ethyl acetate into a separating funnel, shaking the separating funnel, and then removing the first extract with acetic acid. And a method of contacting with ethyl. When the separatory funnel is allowed to stand after contacting the first extract with ethyl acetate, the liquid in the separatory funnel is separated into water and ethyl acetate. The fluorosilenone glycoside of the present invention is in water. Will be included.

[0027] The water collected from the separatory funnel is used as the second extract as it is as the second extract. Use it.

[0028] (3rd step)

In the third step, the second extract obtained in the second step is contacted with butanol. As a result, the fluorosylphenone glycoside of the present invention is selectively incorporated into butanol. An extracted third extract can be obtained. As a method of contacting the second extract with butanol, for example, after the second extract and butanol are placed in a separatory funnel, the separatory port is shaken to remove the second extract from butanol. The method of making it contact with is mentioned. When the separatory funnel is allowed to stand after the second extract is brought into contact with butanol, the liquid in the separatory funnel is separated into water and butanol, but the fluorosilphenone glycoside of the present invention is butanol. Will be included.

[0029] This third step is preferably repeated about 2 to 4 times, in order to obtain more fluorosilanone glycosides.

[0030] The butanol collected from the separatory funnel is a third extract obtained by removing a portion of butanol that may be used as the third extract as it is in the fourth step by evaporation or the like and concentrating it. You may use as a thing.

[0031] (4th step)

In the fourth step, the third extract containing the fluorosylphenone glycoside obtained in the third step is passed through the column packed with the synthetic adsorbent. As a result, the component containing the fluorosilphenone glycoside in the third extract is selectively adsorbed on the synthetic adsorbent as an adsorbing component. Examples of the synthetic adsorbent include Amberlite XAD-4, XAD-7, and XAD-16 (manufactured by Organone, trade name). In addition, other adsorbents such as activated carbon and polypyrrole pyrrolidone (PVPP; polyphenol adsorbent) can be used as the synthetic adsorbent. Among these, XAD-4 is preferably used.

[0032] (5th step)

In the fifth step, the synthetic adsorbent adsorbed by the adsorbing component is brought into contact with a mixed solvent of water and methanol (methanol aqueous solution) to elute the adsorbing component containing the fluorosilphenone glycoside in the mixed solvent. . The mixing ratio of methanol in the mixed solvent is not particularly limited, but is 40 to 60% by mass with respect to the total amount of the mixed solvent from the viewpoint of containing the fluorosilphenone glycoside at a high concentration in the mixed solvent. And preferred. Prior to this fifth step, the synthetic adsorbent with the adsorbed component adsorbed is washed with water or a mixed solvent having a smaller proportion of methanol than the mixed solvent used when eluting the adsorbed component. preferable. In this way, the adsorbed components other than the fluorosilenone glycosides can be synthesized. It can be selectively removed to some extent from the dressing.

[0033] (6th step)

In the sixth step, the column of the high-performance liquid chromatograph (hereinafter referred to as “HPLC”) having a column filled with the above-mentioned mixed solvent containing the adsorbed component eluted in the fifth step and a predetermined adsorbent. Then, separation and purification are performed to isolate the desired fluorosilanphenone glycoside. The isolated fluorosilanone non-glycoside may be appropriately collected according to the retention time. In order to isolate the fluorosilphenone glycoside of the present invention with high purity, it is preferable to use a reverse phase column. Moreover, as the packing material for the column which is a predetermined adsorbent, a porous carrier such as silica gel whose surface is coated with hydrocarbon is preferably used. The hydrocarbon preferably has 8 to 18 carbon atoms. The mixed solvent containing the adsorbed component eluted in the fifth step may pass through any one of these columns, or may pass through two or more columns sequentially.

[0034] Examples of the eluent (mobile phase) include a mixed solution of water and an organic solvent, such as a mixed solution of water and methanol, and a mixed solution of water and acetonitrile. The mixing ratio of each component in the mixed liquid as the eluent is not particularly limited. However, in order to efficiently collect the fluorosilphenone glycoside of the present invention with high purity, water is obtained based on a linear gradient elution method in which the mixing ratio between the two liquids is changed at a constant rate. It is preferable to change the blending ratio of the organic solvent.

[0035] When fractionating the above-mentioned adsorption component force of the fluorosilanone glycoside according to the present invention, each component separated by HPLC is monitored by an analytical method such as an ultraviolet absorption spectrum analysis method. However, it is preferable to take out the fraction.

[0036] Thus, it becomes possible to isolate the novel fluorosylphenone glycoside of the present invention.

[0037] The fluorosilan funon glycoside of the present invention is used for various allergic diseases such as the above-mentioned immediate allergic diseases and chronic allergic inflammations (for example, allergic rhinitis, hay fever, atopic skin). Expected to be used for prevention of symptoms such as inflammation) or relief of symptoms. That is, this novel fluorosilanone glycoside exhibits a function as an antiallergic agent.

[0038] The fluorosilenone glycoside of the present invention has an antiallergic action, in particular, histamine release suppression. Indicates production. For this reason, this fluorosilanone non-glycoside can be used particularly for the purpose of preventing or alleviating symptoms of allergic rhinitis, atopic dermatitis and hay fever. When this fluorosilenone glycoside is added to a pharmaceutical product, it is particularly useful to add it to a preventive or therapeutic agent for allergic rhinitis, atopic dermatitis and hay fever.

[0039] Furthermore, the fluorosylphenone glycoside is used for the purpose of preventing allergic rhinitis, atopic dermatitis and hay fever, and alleviating symptoms, and for specific health foods and drinks, special nutrition foods and drinks, nutritional supplements. It can be added to food additives such as food and drink, health food and drink, functional food and drink, and sick food and drink.

[0040] Further, the structure of the florosylphenone glycoside of the present invention is suggested to exhibit an antioxidant action and a tyrosinase activity inhibitory action. Therefore, when it is added to cosmetics, it can be expected to be added to skin care products, foundations, makeup products, and the like.

[0041] The preferred embodiment of the present invention has been described above, but the present invention is not limited to the above embodiment.

In the preferred embodiment, the first step, the second step, and the third step (hereinafter referred to as “first to third steps”) are provided in this order before the fourth step. However, for example, in another embodiment, the fluorosilenone glycoside can be isolated even if all of the first to fifth steps are omitted. In this case, in the 6th step, instead of the mixed solvent containing the adsorbing component eluted in the 5th step, the cold water extract of the plant tissue is directly passed through the HPLC column including the column filled with the predetermined adsorbent. do it. Alternatively, it is preferable to provide the fourth step and the fifth step before the sixth step because it facilitates the isolation of the fluorosilphenone glycoside. In this case, in the fourth step, the cold water extract of the plant tissue may be directly passed through the column filled with the synthetic adsorbent.

[0043] In the preferred embodiment, the first step and the second step are provided in this order before the third step. However, for example, in another embodiment, either or both of the first step and the second step may be omitted, or these steps may be provided in reverse. In the above preferred embodiment or another embodiment, a cold water extract of plant tissue is added to an organic solvent other than hexane and ethyl acetate before the third step. May be contacted.

[0044] Further, in place of butanol used in the third step, other alcohol having 4 to 5 carbon atoms may be used. However, butanol is preferred in order to contain a fluorosilphenone glycoside with higher selectivity.

[0045] Further, instead of methanol in the mixed solvent used in the fifth step, another alcohol (for example, ethanol) may be used. Furthermore, instead of methanol in the mixed solvent used in the sixth step, another alcohol (for example, ethanol) may be used. Example

Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited to these examples. Unless otherwise specified, “%” represents “volume%”.

[Example 1]

[Isolation of fluorosylphenone glycosides from hop pellets]

Hop pellets (Czech zazat seeds, type 90) lkg was placed in 10 L of distilled water and stirred occasionally at 5 ° C to allow the pellets to stand. After centrifugation at 7000 rpm for 15 minutes, the supernatant (centrifuge supernatant) was recovered. The supernatant was further concentrated to obtain 170 g of concentrate (cold water extract). The concentrate was transferred to a separatory funnel, and hexane was added to the separatory funnel, followed by shaking and standing to remove 2.4 g of the hexane phase from the separatory funnel (first step). Further, ethyl acetate was added to the separating funnel in which the aqueous phase remained, and then shaken and allowed to stand to remove 2.9 g of the ethyl acetate phase from the separating funnel (second step). Finally, n-butanol was added to the separating funnel in which the aqueous phase remained, and then the n-butanol phase was transferred to another container by shaking and standing still (third step). After repeating this third step three times, all n-butanol transferred to another container was collected and concentrated under reduced pressure, and finally 12.2 g of extract was obtained. This extract was confirmed to contain a new fluorosilphenone glycoside represented by the above formulas (2), (3) and (4) by structure determination described later. The remaining aqueous phase in the separatory funnel was 114.6 g.

[0048] In order to further concentrate the fluorosilphenone glycoside to increase its purity, first, a column of 40 mm inner diameter and 450 mm length packed with Amberlite XAD-4 (trade name, manufactured by Organone) as a synthetic adsorbent was packed. The extract was passed through (fourth step). Then extract the After passing through the column, distilled water, 10% aqueous methanol (10% MeOHZH 2 O), 20

2

The column was washed by passing a% methanol aqueous solution (20% MeOHZH 2 O). Then wash

2

Pass the 50% methanol aqueous solution (50% MeOHZH 2 O) through the latter column to contain the extract.

2

3.6 g of an aqueous methanol solution was obtained (fifth step).

[0049] In the fifth step, in order to isolate each component in the aqueous methanol solution obtained in such a manner that it can be separated, the aqueous solution containing the fluorosilenone glycoside is provided in the HPLC. The solution was passed through the column, and each component contained in the aqueous solution was separated (step 6). Two types of columns (first column and second column) were used. As the first column, D-ODS-5 (manufactured by Sakai YMC Co., Ltd., inner diameter 20 mm x length 250 mm, product name) was used at 40 ° C, and the flow rate was 6 mLZ. The eluent (mobile phase) is a linear gradient that uses a mixture of methanol and water, first maintained at 10% methanol for 10 minutes, then changed from 10% methanol to 60% methanol in 160 minutes. . Thus, each component in the aqueous solution was roughly divided. While monitoring with a UV detector (wavelength 290 nm), 240 mg of a solution mainly containing a fluorosilanone glycoside was collected.

[0050] Next, 240 mg of the collected solution was passed through the second column to isolate each component. As the second column, SunFire C18 (Waters, inner diameter 19 mm x length 250 mm, product name) was used at 40 ° C, and the flow rate was 6 mLZ. The eluent (mobile phase) is a linear gradient that uses a mixture of acetonitrile and water, first maintained with 10% acetonitrile for 10 minutes, and then changed the ratio of acetonitrile from 10% acetonitrile to 60% acetonitrile for 160 minutes. . While monitoring with a UV detector (wavelength 290 nm), a total amount of 140 mg of fluorosilenone glycoside was collected. Of these, the compound represented by formula (2) was 110 mg.

[0051] [Structure determination of fluorosilphenone glycoside]

Glycoside components in type 90 hop pellets were identified and determined.

[0052] A cold water extract of type 90 hop pellets was first analyzed by HPLC. In the analysis by HPLC, Symmetry C18 (manufactured by Waters, inner diameter 2. Imm X length 150 mm, trade name) was used as a column at 40 ° C., and the flow rate was 0.2 mLZ. The mobile phase consists of 0.05% TFA ZH O in one solution, acetonitrile in 2 solutions, and a 0.05% TFA / HO ratio of 10% to 50%. The linear gradient was changed to 16% in 16 minutes. Detection was performed with a UV detector (290 nm). Figure 1 shows the HPLC analysis results. From Fig. 1, it was found that six types of components corresponding to peaks A to F mainly exist as glycoside components in the cold water extract of type 90 hop pellets. Of these peaks, A, B, and D were fractionated by the same method as in Step 6 above, and further identified. As a result, isoquercitrin, quercetin malol darcoside and kaempferol malol dalcoside were respectively identified. It turned out to be.

[0053] Next, components corresponding to peak C separated by the same method as in Step 6 above were subjected to ¹ H-NMR, ¹³ C-NMR and ¹ H- H using deuterated methanol! ! -Analyzed by COZY. 11-NMR results are shown in Figs. 2 and 3, ¹³ C-NMR results are shown in Fig. 4, and H-H-COZY results are shown in Fig. 5, respectively.

[0054] Next, a component corresponding to peaking was analyzed using a Waters LCZMS apparatus (MS detector: ZQ detector). Symmetry C18 (manufactured by Waters, ID 2. ImmX length 150 mm, trade name) was used as the LC column at 40 ° C, and the flow rate was 0.2 mLZ. The mobile phase consists of 0.1% formic acid ZH 2 O as one solution, acetonitrile as 2 solutions, and 0.1% formic acid.

2 Z

A linear gradient was used in which the proportion of H 2 O was changed from 10% to 50% in 16 minutes. UV detection

2

The monitoring was carried out with an ejector (290 nm). Fig. 6 (a) shows the result of positive scan (mZzl20 to m Zz650, fixed voltage: + 3. OkV, cone voltage: + 25V), and negative scan (m / zl20 to m / z650, fixed voltage: — 3. OkV, cone voltage: (25V) The results are shown in Fig. 6 (b). From this result, the molecular ion was cleaved and a small signal of mZzl62 was detected, which revealed that the glycoside is a hexose.

[0055] Next, 7.5 mg of the component corresponding to peak C was dissolved in 13 mL of 2N HC, and refluxed at 100 ° C for 30 minutes for hydrolysis. Thereafter, the obtained reaction solution was passed through a column packed with MEGAB OND ELUT C18 (2 g, manufactured by Varian, trade name) as an adsorbent, the first 10 mL was discarded, and the next 10 mL was recovered. About the collected reaction liquid NH column (

2 LCZMS analysis was performed using a product name “CAPCELL PAK NH2 UG80” (inner diameter 2. Omm × length 25 Omm) manufactured by Shiseido Co., Ltd. The analysis conditions were a column temperature of 80 ° C, an eluent of 89% acetononitrile Z water at a flow rate of 0.25 mLZ, and negative SIR (mZzl79, key (Capillary voltage: 3.2 kV, cone voltage: 25 V). For reference, the same analysis was performed for mannose, galactose and glucose. Figure 7 shows the resulting mass spectrum. (A) is mannose, (b) is galactose, (c) is gnolecose, and (d) is the mass spectrum of the recovered reaction solution. This revealed that the constituent sugar of the glycoside was glucose, and it was confirmed that the component corresponding to peak C was a compound represented by formula (2).

[0056] Next, UV absorption of components corresponding to peak C, peak E, and peak F in the HPLC chromatogram in Fig. 1 was measured. The resulting UV absorption spectrum is shown in Fig. 8. (A) is the peak C, (b) is the peak E, and (c) is the UV absorption spectrum of the component corresponding to the peak F, respectively. As a result, the UV absorption spectrum pattern was the same for all three components.

[0057] Next, LCZMS analysis of components corresponding to peak E and peak F was performed. Fig. 14 shows the results of LCZMS analysis performed under the same conditions as those for obtaining the mass spectrum of Fig. 6 (a). Fig. 15 shows the same conditions as those for obtaining the mass spectrum of Fig. 6 (b). The results of L CZMS analysis performed in the above are shown. 14 and 15, (a) is the mass spectrum of the component corresponding to the peak E and (b) is the component corresponding to the peak F, respectively. From this result, it was found that the components corresponding to peak E and peak F were!, And the deviation was 14 larger than the component corresponding to peak C. This result suggests that the component corresponding to peak E and peak F has the same basic skeleton as the component corresponding to peak C, and is different only in the side chain structure.

[0058] Finally, components corresponding to peak E and peak F were analyzed by ¹ H-NMR, ¹³ C-NMR and ¹ H- ¹ H-COZY using deuterated methanol. Fig. 9 shows the results of ¹ H-N MR of the component corresponding to peak F, Fig. 10 shows the results of ¹³ C-NMR, and Fig. 11 shows the results of ipi- ¹ !!-COZY. The result of ¹ H-NMR of the component corresponding to peak E is shown in FIG. 12, and the result of 'Η-'Η-COZY is shown in FIG. From these results, it was confirmed that the component corresponding to peak Ε was a compound represented by formula (4), and the component corresponding to peak F was a compound represented by formula (3).

[Example 2] [In vitro assay to confirm histamine release inhibitory effect of fluorosylphenone glycosides]

Human basophil cell line (KU812) was cultured in RPMI1640 medium (Gibco) containing fetal calf serum that had been inactivated for 10 minutes at 56 ° C at 37 ° C in a 5% CO atmosphere. Cultured.

2

Add Tyrode solution to the cells, centrifuge (1500 rpm, 5 minutes, 4 ° C), repeat washing twice with Tyrode solution, suspend in Tyrode solution, and add 2 X to a 5 mL tube. 10 ⁶ cell sZmL was dispensed. Each cell suspension contains the amount of buffer shown in Table 1, 10 mM CaCl, 50 M A23187 and

2 Prepare Z or a test compound (fluorosylphenone glycoside represented by formula (2)), allow histamine release reaction at 37 ° C for 20 minutes, and then put in ice for 5 minutes to react. Stopped.

[0060] [Table 1]

[0061] Thereafter, the mixture was centrifuged at 4 ° C for 3 minutes at lOOOOrpm, and the supernatant was collected. Collected supernatant force Extracting free histamine with an organic solvent, and o Exciting the fluorescence generated by reaction with phthalaldehyde by measuring the fluorescence intensity at a wavelength of 450 nm after excitation with light of a wavelength of 350 nm. Histamine was quantified. The total intracellular histamine was measured by measuring the histamine content in the supernatant obtained by sonicating the same amount of cell suspension in ice for 1 minute and then centrifuging at 4 ° C for 3 minutes at lOOOOrpm. Was obtained by The inhibition rate of histamine release is determined by the formula: inhibition rate of histamine release (%) = (histamine content in the supernatant of each sample, natural release amount) X 100 Z (histamine release amount by A23187 stimulation-natural release amount) Based on the results, the IC50 value (50% inhibitory concentration) of the fluorosilenone glycoside represented by the formula (2) was determined. As a result, the IC50 value was 290 g / mL.

Industrial applicability

[0062] According to the present invention, the antiallergic action is sufficiently excellent and the side effects are sufficiently suppressed. Can provide substances that are mild to the human body and skin.

Claims

Claims [1] A fluorosilanone glycoside represented by the following general formula (1). [Chemical 1]

[In the formula (1), R ¹ represents an isopropyl group, an isobutyl group or a sec-butyl group. ]

[2] The fluorosylphenone glycoside according to claim 1, obtained by separation from a cold water extract of a plant tissue.

[3] The fluorosilphenone glycoside according to claim 1 or 2, wherein the plant tissue is a hop tissue.

[4] The floracylphenone glycoside according to claim 3, wherein the hop tissue is a stem, a spike or a leaf.

[5] The fluorosilanone non-glycoside according to any one of claims 1 to 4, which exhibits an antiallergic action.

[6] The fluorosilenone glycoside according to any one of claims 1 to 4, which exhibits an action as an antioxidant.

[7] The fluorosylphenone glycoside according to any one of claims 1 to 4, which exhibits a tyrosinase activity inhibitory action.

[8] Food and drink containing the fluorosilphenone glycoside according to any one of claims 1 to 7

P

PPo

[9] A cosmetic comprising the fluorosilphenone glycoside according to any one of claims 1 to 7.

P

PPo

[10] A medicament comprising the fluorosilenone glycoside according to any one of claims 1 to 7.