JP2005272374A - Flavanone compound, its manufacturing method and antioxidant - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new flavanone compound which exhibits a high anti-oxidizing action, its manufacturing method, and an antioxidant. <P>SOLUTION: The flavanone compound has a structure represented by formula (1), formula (2) or formula (3). The flavanone compound can be obtained by using propolis including Macaranga tanarius Muell. Arg. of the origin plant as the raw material, and performing an extraction step of extracting an extract component containing the flavanone compound and then a separation step of separating the flavanone compound from the extract component. The flavanone compound having a structure represented by formula (3) can be obtained by using Macaranga tanarius Muell. Arg. as the raw material and performing the above extraction step and separation step. The antioxidant contains the above flavanone compound as an active ingredient. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a novel flavanone compound, a production method thereof and an antioxidant containing the flavanone compound.

Conventionally, nymphaeol-A, B, and C are known as this kind of flavanone compound (for example, refer nonpatent literature 1).
K. Yakushijin, K. Shibayama, H. Murata and H. Furukawa, New prenylflavanones from Hernandia nymphaefolia (presl) Kubitzki, Heterocycles, 14, 397-402, 1980.

  The present invention has been made by isolating a novel flavanone compound and finding a useful physiological activity as a result of intensive studies by the present inventors. The object is to provide a novel flavanone compound that exhibits a high antioxidant action, a method for producing the same, and an antioxidant.

  In order to achieve the above object, the flavanone compound of the invention according to claim 1 has a structure represented by the following chemical formula (1).

請求項２に記載の発明のフラバノン化合物は、下記化２に示される構造を有するものである。

The flavanone compound of the invention described in claim 2 has a structure shown in the following chemical formula 2.

請求項３に記載の発明のフラバノン化合物は、下記化３に示される構造を有するものである。

The flavanone compound of the invention described in claim 3 has a structure shown in the following chemical formula 3.

請求項４に記載の発明のフラバノン化合物の製造方法は、請求項１から請求項３のいずれか一項に記載のフラバノン化合物の製造方法であって、起源植物にオオバギを含むプロポリスを原料として前記フラバノン化合物を精製するものである。

A method for producing a flavanone compound according to a fourth aspect of the present invention is the method for producing a flavanone compound according to any one of the first to third aspects, wherein a propolis containing a grasshopper is used as a raw material. It purifies a flavanone compound.

  A method for producing a flavanone compound according to a fifth aspect of the present invention is the method for producing a flavanone compound according to the third aspect, wherein the flavanone compound is purified using a grasshopper as a raw material.

  The antioxidant of invention of Claim 6 contains the flavanone compound of Claim 1, Claim 2, or Claim 3.

  According to the flavanone compound of the present invention, a high antioxidant effect can be exhibited. According to the method for producing a flavanone compound of the present invention, the flavanone compound can be easily produced. According to the antioxidant of the present invention, a high antioxidant action can be exhibited.

  The first flavanone compound of the embodiment is 5,7,3 ′, 4′-tetrahydroxy-6- (7 ″ -hydroxy-3 ″, 7 ″ -dimethyl- having the structure shown in the following chemical formula 4. oct-2 ″ -enyl) -flavanone.

この第１のフラバノン化合物は、分子式Ｃ₂₅Ｈ₃₀Ｏ₇、分子量４４２、融点（ＭＰ）９７〜１００℃である（図１参照）。このフラバノン化合物は新規化合物であり、エリオディクティオール（Eriodictyol）と類似した構造的特徴を有しているが、６位に7''-hydroxy-3'',7''-dimethyl-oct-2''-enyl基を備えていることからエリオディクティオールよりも親油性が高い。

This first flavanone compound has a molecular formula C ₂₅ H ₃₀ O ₇ , a molecular weight 442, and a melting point (MP) 97-100 ° C. (see FIG. 1). This flavanone compound is a new compound and has structural characteristics similar to those of Eriodictyol, but 7 ''-hydroxy-3 '', 7 ''-dimethyl-oct- Because it has a 2 ''-enyl group, it is more lipophilic than Eriodictiool.

  The second flavanone compound of the embodiment is 5,7,4′-trihydroxy-3 ′-(7 ″ -hydroxy-3 ″, 7 ″ -dimetyl-oct- having the structure shown in the following chemical formula 5 2 ″ -enyl) -flavanone.

この第２のフラバノン化合物は、分子式Ｃ₂₅Ｈ₃₀Ｏ₆、分子量４２６、融点７１〜７４℃である（図３参照）。このフラバノン化合物は新規化合物であり、ナリンゲニン（Naringenin）と類似した構造的特徴を有しているが、３’位に7''-hydroxy-3'',7''-dimetyl-oct-2''-enylを備えていることからナリンゲニンよりも親油性が高い。

This second flavanone compound has a molecular formula of C ₂₅ H ₃₀ O ₆ , a molecular weight of 426, and a melting point of 71 to 74 ° C. (see FIG. 3). This flavanone compound is a novel compound and has structural characteristics similar to that of Naringenin, but 7′-hydroxy-3 ″, 7 ″ -dimetyl-oct-2 ′ is located at the 3 ′ position. Because it has' -enyl, it is more lipophilic than naringenin.

  The third flavanone compound of the embodiment is 5,7,4′-trihydroxy-3′-geranylflavanone having the structure shown in the following chemical formula 6.

この第３のフラバノン化合物は、分子式Ｃ₂₅Ｈ₂₈Ｏ₅、分子量４０８、融点４８〜５０℃である（図５参照）。このフラバノン化合物は新規化合物であり、ナリンゲニンと類似した構造的特徴を有しているが、３’位にＣ−ゲラニル基を備えていることからナリンゲニンよりも親油性が高いうえ、エリオディクティオールよりも親油性が高い。

This third flavanone compound has a molecular formula of C ₂₅ H ₂₈ O ₅ , a molecular weight of 408, and a melting point of 48 to 50 ° C. (see FIG. 5). This flavanone compound is a novel compound and has structural characteristics similar to naringenin, but has a lipophilicity higher than naringenin because it has a C-geranyl group at the 3 ′ position. Higher lipophilic than.

  The first to third flavanone compounds have a high antioxidant effect. Regarding the antioxidant action, the first flavanone compound is particularly high, and the second and third flavanone compounds are slightly lower than the first flavanone compound. Each of the first to third flavanone compounds is obtained by using a propolis containing plantain as a raw material, and performing an extraction process for extracting an extraction component containing the flavanone compound from the raw material, and then separating the flavanone compound from the extraction component It is obtained by performing a separation step. Furthermore, the third flavanone compound can also be obtained by performing the extraction step and the separation step using Euobagi as a raw material. The grasshopper is also known as Macaranga tanarius Muell. Arg. It is an evergreen broad-leaved tree (male and female) belonging to the Euphorbiaceae. Weow grows in Okinawa, Taiwan, southern China, the Malay Peninsula, Southeast Asia such as the Philippines, Malaysia, Indonesia, and Thailand, and northern Australia.

  The above-mentioned propolis may be composed only of a grasshopper plant, or a combination of a grasshopper and another plant (Bidens pilosa L. var. Minor Scheff., Citrus depressa Hay, etc.) May be configured. Propolis production areas include areas where weow grows, that is, Okinawa, Taiwan, etc. Okinawa is preferred because of the high content of flavanone compounds. The raw material of the flavanone compound may be composed of a propolis derived from a single production area, or may be composed of a propolis mixture derived from a plurality of production areas. On the other hand, when a plantain is used as a raw material for the third flavanone compound, the raw material is composed of various parts such as leaf blade, petiole, stem, stem, fruit, etc., but the content of the third flavanone compound is high. In addition, a leaf blade or a petiole is preferable.

  In the extraction step, the extraction component is extracted from the raw material by immersing the raw material in an extraction solvent or performing supercritical extraction. Examples of the extraction solvent include methanol, ethanol, acetonitrile, ethyl acetate, hexane, water, and the like. However, methanol or ethanol is preferable and ethanol is more preferable because extraction efficiency of extraction components is high. In the separation step, the flavanone compound is purified or isolated from the extracted component by a method of fractionating the extracted component by chromatography using a silica gel carrier or the like.

  The antioxidant of the embodiment contains the first, second or third flavanone compound as an active ingredient (antioxidant material). Antioxidants are food / beverage products (beverages) that effectively prevent deterioration of various products (mainly oxidative deterioration) such as oxidative degradation of fats and oils, perfume degradation, pigment degradation, and pigment fading. Alternatively, it can be used by being added in food). In addition, antioxidants can be used in health foods and other foods and drinks to eliminate active oxygen in the body when taken orally, enhance liver function, reduce toxicity of acetaldehyde, low density It exerts health promotion effects such as antioxidant action of cholesterol (LDL). Furthermore, the antioxidant can be used by being incorporated in cosmetics, pharmaceuticals or quasi-drugs, and is useful for the whitening effect of skin, oral cavity, etc., prevention of aging, and the like.

  The food / beverage products of embodiment contain the said 1st, 2nd, or 3 flavanone compound, and may contain the said antioxidant. The food and drink can be utilized as a health food that exhibits various health promoting effects by eliminating active oxygen in vivo by sufficiently drawing out the antioxidant action of the flavanone compound. In addition, by sufficiently drawing out the degradation preventing action of the flavanone compound, it is possible to prevent deterioration of the food and drink, increase the storage stability, and maintain a stable quality for a long period of time. In food and drink, the daily intake of the flavanone compound is preferably 0.01 to 10 g, more preferably 0.2 to 5 g per day for an adult. If the daily intake of the flavanone compound is less than 0.01 g, the antioxidant action may not be exhibited effectively, and conversely if it exceeds 10 g, it is uneconomical. In the case of a dwarf, half the amount for an adult is a guide.

The effects exhibited by the above embodiment will be described below.
-The first to third flavanone compounds of the embodiments all have an antioxidant action. For this reason, each flavanone compound can be used for various uses using the fact that it is more lipophilic than Eriodictiool and Naringenin, in addition to being able to be used for the same applications as Eriodictiool and Naringenin. Further, since each flavanone compound is contained in propolis used as a health food material, there is no problem in oral intake and transdermal administration. In addition, since each flavanone compound has a high antiallergic action, antibacterial action and antitumor action in addition to the antioxidant action, it can be used as an active ingredient of an antiallergic agent, antibacterial agent or antitumor agent. it can.

  -In the manufacturing method of the 1st and 2nd flavanone compound of embodiment, the origin plant uses the propolis which contains a grass as a raw material, and the 3rd flavanone compound manufacturing method uses the origin plant as a propolis or a grasshopper. Propolis contains the first to third flavanone compounds, and the weow contains the third flavanone compounds. For this reason, according to each manufacturing method, manufacture of each flavanone compound is easy.

  The grasshopper contains almost no first and second flavanone compounds, but contains nymphaeol-A (5,7,3 ', 4'-tetrahydroxy-6-geranylflavanone) and third flavanone compounds. (See FIG. 8). Here, the first flavanone compound has a structure in which the terminal of the alkyl chain in the C-geranyl group of Nimpheol-A is hydroxylated, and the second flavanone compound is the C-geranyl group of the third flavanone compound. In which the terminal of the alkyl chain is hydroxylated.

  -Since the antioxidant of the embodiment contains a flavanone compound having a high antioxidant action as an active ingredient, it prevents deterioration of foods, drinks, cosmetics, pharmaceuticals or quasi-drugs and enhances storage stability, By ingestion or transdermal administration, health enhancement effect and anti-aging effect can be exhibited. Moreover, since the food / beverage products of embodiment contain the flavanone compound which has an antioxidant effect | action, the deterioration prevention and health promotion effect of food / beverage products themselves can be exhibited.

Next, the embodiment will be described more specifically with reference to test examples and comparative examples.
<Isolation of compound>
An extraction process was performed by adding 500 ml of ethanol to 50 g of propolis raw material from Okinawa, sonicating for several minutes and stirring overnight at room temperature (25 ° C.), followed by filtration to remove the residue. Here, as the above-mentioned Okinawa propolis body, one having Naha City, Okinawa Prefecture as the production area was used. Next, the obtained extract was concentrated under reduced pressure to obtain 39.73 g of an ethanol extract. Subsequently, as a separation step, the ethanol extract was fractionated into 11 fractions (1) to (11) by column chromatography under the following conditions.

Column tube: Glass column 5.0 x 45 cm
Filler: Silica gel about 590cm ³
Elution solvent: (1) Hexane: ethyl acetate = 90: 10 (350 ml)
(2) Hexane: ethyl acetate = 80: 20 (220 ml)
(3) Hexane: ethyl acetate = 70: 30 (250 ml)
(4) Hexane: ethyl acetate = 60: 40 (1000 ml)
(5) Hexane: ethyl acetate = 50: 50 (200 ml)
(6) Hexane: ethyl acetate = 40: 60 (100 ml)
(7) Hexane: ethyl acetate = 30:70 (100 ml)
(8) Hexane: ethyl acetate = 20: 80 (100 ml)
(9) Hexane: ethyl acetate = 10:90 (100 ml)
(10) Ethyl acetate (200ml)
(11) Methanol (700ml)
Next, when each fraction was analyzed under the following HPLC condition 1, it was confirmed that the fractions (4) and (6) to (9) contained a total of nine main components.

HPLC condition 1
Column: Shiseido Capcell Pak ODS UG-120 (4.6 × 150mm)
Solvent: A: Water (2% acetic acid), B: Acetonitrile (2% acetic acid)
Elution condition: 0-60min (gradient elution; A: B = 80: 20 → A: B = 20: 80)
Flow rate: 1ml / min
Detection: UV280nm
Next, fractionation was carried out under the following HPLC condition 2 using the fractions (6) to (9) to isolate compounds 1, 2, 3 and 4. Further, the compounds 1 and 2 were purified under the following HPLC condition 3, and the compounds 3 and 4 were purified under the following HPLC condition 4. The yield of Compound 1 after purification was 73.1 mg, the yield of Compound 2 was 8.6 mg, the yield of Compound 3 was 9.1 mg, and the yield of Compound 4 was 13.4 mg. In addition, the following HPLC conditions 3 and 4 show only conditions different from the HPLC condition 2, respectively.

HPLC condition 2
Column: YMC-Pack R & D ODS (20 × 250mm)
Solvent: Water (0.1% TFA): Acetonitrile (0.1% TFA) = 45: 55
Flow rate: 9ml / min
Detection: UV280nm
HPLC condition 3
Solvent: Water (0.1% TFA): Acetonitrile (0.1% TFA) = 50: 50
Flow rate: 8ml / min
HPLC condition 4
Solvent: Water (0.1% TFA): Acetonitrile (0.1% TFA) = 40:60
In addition, fractionation was performed using the fraction (4) under the following HPLC condition 5 to isolate compounds 5, 6, 7, 8, and 9. Further, the compounds 5 and 6 were purified under the following HPLC condition 6, the compounds 7 and 8 were purified under the following HPLC condition 7, and the compound 9 was purified under the following HPLC condition 8. The yield of compound 5 after purification was 307.1 mg, the yield of compound 6 was 231.5 mg, the yield of compound 7 was 96.8 mg, the yield of compound 8 was 101.0 mg, The yield was 142.0 mg. In addition, the following HPLC conditions 6-8 show only conditions different from the HPLC condition 5, respectively.

HPLC condition 5
Column: YMC-Pack R & D ODS (20 × 250mm)
Solvent: Water (0.1% TFA): Acetonitrile (0.1% TFA) = 40:60
Flow rate: 9ml / min
Detection: UV280nm
HPLC condition 6
Solvent: Water (0.1% TFA): Acetonitrile (0.1% TFA) = 35: 65
HPLC condition 7
Solvent: Water (0.1% TFA): Acetonitrile (0.1% TFA) = 35: 65
Flow rate: 8ml / min
HPLC condition 8
Solvent: Water (0.1% TFA): Acetonitrile (0.1% TFA) = 20:80
<Identification of each compound>
Each of the compounds 3, 4 and 8 was subjected to structural analysis by measuring ¹ H-NMR, ¹³ C-NMR, MS, IR, UV spectrum and the like. Details are described below. In addition, when a structural analysis was performed in the same manner for compounds other than the above-mentioned compounds, Compound 1 was 5,7,3 ', 4'-tetrahydroxy-2'-(7 ''-hydroxy-3 '', 7 ''-dimethyl-oct-2''-enyl) -flavanone and compound 2 is 5,7,3', 4'-tetrahydroxy-5 '-(7''-hydroxy-3'',7''- Dimethyl-oct-2 ″ -enyl) -flavanone, and Compound 5 was found to be 5,7,3 ′, 4′-tetrahydroxy-2′-geranylflavanone. Further, Compound 6 is 5,7,3 ′, 4′-tetrahydroxy-5′-geranylflavanone, Compound 7 is Nimpheol-A, and Compound 9 is 5,7,3 ′, 4′-tetrahydroxy-6- It was found to be (3 ''',3'''-dimethylallyl)-2'-geranylflavanone.

(Compound 3: Compound 3)
The physicochemical properties are shown in FIG. Molecular weight from measurement by ESI-MS is identified as 442, a hydroxyl group, the presence of carbonyl group 1680 cm ^-1 was suggested to 3430cm ^-1 from the IR spectrum. In addition, in the UV spectrum, a characteristic spectrum of flavanone or flavanol was shown, so that it was presumed to have a flavanone skeleton. Thirty protons were confirmed from the integrated value of the ¹ H-NMR spectrum, and 24 signals were observed in the ¹³ C-NMR spectrum. From the DEPT spectrum, 3 methyls, 5 methylenes, 6 methines and 11 quaternary carbons were confirmed. From these information, the molecular formula was estimated as C ₂₅ H ₃₀ O ₇ . In addition to these, NMR data revealed from HSQC spectrum, ¹ H- ¹ H COSY and HMBC spectrum are summarized in FIG. From the above results and the results of CD spectrum, this compound has the structure shown in Chemical Formula 4, and 5,7,3 ', 4'-tetrahydroxy-6- (7''-hydroxy-3'' , 7 ''-dimethyl-oct-2 ''-enyl) -flavanone. This compound was a novel compound not yet described in literatures.

(Compound 4: Compound 4)
The physicochemical properties are shown in FIG. Molecular weight from measurement by ESI-MS is identified as 426, a hydroxyl group, the presence of carbonyl group 1680 cm ^-1 was suggested to 3430cm ^-1 from the IR spectrum. In addition, in the UV spectrum, a characteristic spectrum of flavanone or flavanol was shown, so that it was presumed to have a flavanone skeleton. Thirty protons were confirmed from the integrated value of the ¹ H-NMR spectrum, and 24 signals were observed in the ¹³ C-NMR spectrum. From the DEPT spectrum, 3 methyls, 5 methylenes, 7 methines and 10 quaternary carbons were confirmed, and the molecular formula was estimated to be C ₂₅ H ₃₀ O ₆ from these information. In addition to these, the NMR data clarified from the HSQC spectrum, ¹ H- ¹ H COSY and HMBC spectrum are summarized in FIG. From the above results and the results of CD spectrum, this compound has the structure shown in Chemical Formula 5, and 5,7,4'-trihydroxy-3 '-(7''-hydroxy-3'', 7 It was identified as '' -dimetyl-oct-2 ''-enyl) -flavanone. This compound was a novel compound not yet described in literatures.

(Compound 8)
The physicochemical properties are shown in FIG. Molecular weight from measurement by ESI-MS is identified as 408, a hydroxyl group, the presence of carbonyl group 1680 cm ^-1 was suggested in 3400 cm ^-1 from the IR spectrum. In addition, in the UV spectrum, a characteristic spectrum of flavanone or flavanol was shown, so that it was presumed to have a flavanone skeleton. ^From the integrated value of the ¹ H-NMR spectrum, 28 protons were confirmed, and in the ¹³ C-NMR spectrum, 25 signals were observed. From the DEPT spectrum, 3 methyls, 4 methylenes, 8 methines, and 10 quaternary carbons were confirmed. From these information, the molecular formula was estimated as C ₂₅ H ₂₈ O ₅ . In addition to these, NMR data revealed from HSQC spectrum, ¹ H- ¹ H COSY and HMBC spectrum are summarized in FIG. From the above results and CD spectrum results, this compound had the structure shown in Chemical Formula 6 and was identified as 5,7,4′-trihydroxy-3′-geranylflavanone. This compound was a novel compound not yet described in literatures.

<DPPH radical scavenging activity test>
DPPH (α, α-diphenyl-β-picrylhydradil) is a purple stable radical having a maximum absorption at 517 nm, and turns into colorless hydrazine by obtaining hydrogen. Utilizing this color reaction, radical scavenging activity was measured by the following method. That is, the compound 3, 4 or 8 as a sample was dissolved in ethanol to prepare 0.5 ml of a sample solution. Subsequently, 0.5 ml of a 0.15 mM DPPH solution (the solvent was ethanol) was added to each sample solution, stirred, and allowed to react in the dark for 1 hour, and then the absorbance at 517 nm was measured. On the other hand, as a comparative control, the same test was conducted using BHT (butylated hydroxytoluene), α-tocopherol or eriodictiool as a sample instead of the compound. Here, the final concentration of the sample was 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, or 100 μM. Moreover, the test was similarly implemented using the thing which has not added the said sample as a control. The radical scavenging activity (%) was calculated by the following formula 1. Furthermore, the value of EC ₅₀ (μM) was calculated from the value of radical scavenging activity at each final concentration. The results are shown in Table 1 below (all tests were performed in triplicate, showing the mean and standard deviation).

表１に示すように、３種類の化合物とも各比較対照と同様にラジカル捕捉活性を示した。化合物３のラジカル捕捉活性は高く、そのＥＣ₅₀がＢＨＴやα−トコフェロールよりも低い値を示しエリオディクティオールとほぼ同等であることが明らかとなった。

As shown in Table 1, the three compounds showed radical scavenging activity as in the case of each comparative control. The radical scavenging activity of Compound 3 was high, and its EC ₅₀ was lower than that of BHT and α-tocopherol, and was found to be almost equivalent to Eriodictiool.

<Β-Carotene fading test>
This method utilizes a phenomenon in which the color of β-carotene disappears by the reaction of the linoleic acid peroxide generated with auto-oxidation of linoleic acid with the double bond of β-carotene. Antioxidant activity was measured by the method described above. Specifically, after mixing 2.0 ml of 0.2 g / ml Tween 40 chloroform solution, 0.4 ml of 0.1 g / ml linoleic acid chloroform solution, and 3.0 ml of 0.1 mg / ml β-carotene chloroform solution, The solvent was removed using nitrogen gas. Subsequently, 100 ml of distilled water was added and stirred sufficiently to obtain an emulsion. On the other hand, the compound 3, 4 or 8 as a sample was dissolved in ethanol to prepare 50 μl of a sample solution. Next, 3 ml of the emulsion was added to each sample solution to prepare a reaction solution, and the reaction solution was incubated at 60 ° C. for 60 minutes. Absorbance at 470 nm was measured for the reaction solution before incubation (sample reaction solution of 0 minutes) and the reaction solution after incubation (sample reaction solution of 60 minutes). Here, the final concentration of the sample was 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, 100 μM, or 200 μM.

On the other hand, as a comparative control, a reaction solution using BHT, α-tocopherol or eriodictiool as a sample instead of the above compound was prepared and similarly tested. Moreover, the control reaction liquid which did not add the said sample was prepared, and the test was implemented similarly. Antioxidant activity (%) was determined by the following formula 2. Furthermore, the value of IC ₅₀ (μM) was calculated from the value of the antioxidant activity at each final concentration. The results are shown in Table 2 below (all tests were performed in triplicate, showing the mean and standard deviation).

但し、前記コントロールの退色速度は、（０分のコントロール反応液の吸光度／０分の試料反応液の吸光度）／６０の値の自然対数で表され、前記試料の退色速度は、（６０分のコントロール反応液の吸光度／６０分の試料反応液の吸光度）／６０の値の自然対数で表される。

However, the color fading rate of the control is represented by the natural logarithm of the value of (absorbance of the control reaction solution of 0 minute / absorbance of the sample reaction solution of 0 minute) / 60. The absorbance of the control reaction solution / absorbance of the sample reaction solution for 60 minutes) / the natural logarithm of the value of 60.

表２に示すように、３種類の化合物とも各比較対照と同様に抗酸化活性を示すことが明らかとなった。一方、化合物８についてリポソームへの取込み量を測定したところ、データは示さないがエリオディクティオールよりも取込み量が多い結果となった。

As shown in Table 2, it was clarified that all the three types of compounds exhibited antioxidant activity as in the case of each comparative control. On the other hand, when the amount of compound 8 incorporated into the liposome was measured, the data was not shown, but the amount incorporated was larger than that of Eriodictiool.

<Component analysis of propolis drug substance by production area>
A sample solution (concentration of propolis drug substance: 2 mg / ml) was prepared by adding ethanol to the propolis drug substance and performing an extraction process. Next, the sample solution was filtered (filter pore size: 0.45 μm) to remove the residue, and then analyzed under the following HPLC condition 9. Here, as the propolis base, in addition to the Okinawa propolis base, Hokkaido Kawakami-gun, Minami-Akita-gun, Akita-ken, Kazuno-shi, Akita-ken, Aizuwakamatsu-shi, Fukushima-ken, Futaba-gun, Fukushima-ken, Toki-shi, Gifu, Nagano Prefecture Matsumoto City, Tokyo Machida City, Kanagawa Prefecture Ashigara County, Shizuoka Prefecture Takata County, Okayama Prefecture Kamata County, Tottori Prefecture Kurayoshi City or Fukuoka Prefecture Yame County, Japan, Urushi Valley, Cheongju, Imasa, Moju , Korea made from Pocheon or Sangju, or China (Hebei). The HPLC chromatogram of the propolis raw material from Okinawa is shown in FIG. 7, and the legend in FIG. 7 is shown below. On the other hand, the display of the HPLC chromatogram is omitted for propolis active substances other than Okinawa propolis active substances.

HPLC condition 9
Column: Shiseido Capcell Pak C18 UG-120 (4.6 × 250mm)
Solvent: A: Water (2% acetic acid), B: Acetonitrile (2% acetic acid)
Elution condition: 0-60min (A: B = 20: 80)
Flow rate: 1ml / min
Detection: UV280nm
Injection volume: 5 μl
Temperature: 30 ° C
(Legend)
1: 5,7,3 ', 4'-tetrahydroxy-2'-(7 ''-hydroxy-3 '', 7 ''-dimethyl-oct-2 ''-enyl) -flavanone
2: 5,7,3 ', 4'-tetrahydroxy-5'-(7 ''-hydroxy-3 '', 7 ''-dimethyl-oct-2 ''-enyl) -flavanone
3: Compound 3 (first flavanone compound)
4: Compound 4 (second flavanone compound)
5: 5,7,3 ', 4'-tetrahydroxy-2'-geranylflavanone
6: 5,7,3 ', 4'-tetrahydroxy-5'-geranylflavanone
7: Nimpheol-A
8: Compound 8 (third flavanone compound)
9: 5,7,3 ', 4'-tetrahydroxy-6- (3''', 3 '''-dimethylallyl)-2'-geranylflavanone
As a result, as shown in FIG. 7, the propolis raw material produced in Okinawa has a composition different from that of the propolis raw material derived from poplar or the propolis raw material derived from baccaris, although no data is shown. It became clear that. On the other hand, the propolis drug substance other than the propolis drug substance produced in Okinawa has a similar composition to the propolis drug substance originating from poplar, although no data is shown.

<Plant component analysis>
We collected leaves, stems, trunks and fruits in April in Shuri Kinjo-cho, Naha City, Okinawa, and then chopped them together and crushed them in a mortar to obtain samples. Next, ethanol was added at a rate of 1 ml per 0.1 g of the sample, and the extract was left in a dark place at room temperature for about 1 week to obtain an extract. Subsequently, after the extract was dried to obtain an extract, ethanol was added to prepare a sample solution (concentration of extract: 20 mg / ml). Next, the sample solution was filtered (pore size of the filter: 0.45 μm) to remove the residue, and then analyzed under the following HPLC condition 10. In addition, the following HPLC conditions 10 show only conditions different from HPLC conditions 9. On the other hand, analysis was carried out in the same manner as described above using white ginseng, Sikhwasa or Okinawa propolis drug substance in place of the above-mentioned grasshopper. Each HPLC chromatogram is shown in FIG. In addition, each number in FIG. 8 shows the same legend as FIG. 7, P is an Okinawa propolis raw material, a is a weeping, b is white ginseng, and c is a seeker.

HPLC condition 10
Injection volume: 10 μl
As a result, as shown in FIG. 8, it was clarified that compound 8 was contained in the grass, and compounds 1 to 9 were not contained in white ginseng and seeker. Furthermore, it was revealed that the HPLC chromatogram of the propolis from Okinawa was similar only to the HPLC chromatogram of the grasshopper. Here, weows grew in the vicinity of the apiary where the propolis from Okinawa was collected. For this reason, it is highly possible that the propolis of Okinawa origin contains a grasshopper in the plant of origin.

<Ingredient analysis of each part of the grasshopper>
In October, in Okinawa Prefecture, Shuri Kinjo Town, wean leaves, stem tip, petiole, stem (excluding tip) and stem were collected. Next, except that the stems and stems were combined into one, each part was divided separately, and each was finely chopped and ground in a mortar to obtain a sample derived from a specific part. Subsequently, each sample was analyzed in the same manner as in the above <analysis of plant components>. On the other hand, instead of each part of the grasshopper, analysis was performed using a sample (site mixture) derived from the grasshopper in <Plant component analysis> or a propolis raw material from Okinawa. Each HPLC chromatogram is shown in FIG. In addition, each number in FIG. 9 shows the same legend as FIG. 7, P is Okinawa propolis body, a is a mixture of parts, b is leaf blades, c is the tip of stem, d is petiole, e is stem and stem A mixture of As a result, as shown in FIG. 9, it was revealed that the content of Compound 8 was higher in the leaf blades and petioles than in other parts.

The physicochemical properties of Example 3 were summarized. The NMR data of the compound 3 of an Example are shown. The physicochemical properties of Example 4 were summarized. The NMR data of the compound 4 of an Example are shown. The physicochemical properties of Example 8 were summarized. The NMR data of the compound 8 of an Example are shown. The HPLC chromatogram of the propolis raw material from Okinawa is shown. The HPLC chromatogram of each plant is shown. The HPLC chromatogram of each site | part in a grasshopper is shown.

Claims (6)

The flavanone compound which has a structure shown by following Chemical formula 1.

The flavanone compound which has a structure shown by following Chemical formula 2.

The flavanone compound which has a structure shown by following Chemical formula 3.

4. A method for producing a flavanone compound according to any one of claims 1 to 3, wherein the flavanone compound is purified using a propolis containing a grass as a starting material. . 4. The method for producing a flavanone compound according to claim 3, wherein the flavanone compound is purified using green grass as a raw material. The antioxidant containing the flavanone compound of Claim 1, Claim 2, or Claim 3.

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