JP2002047196A - Fruit polyphenol, antioxidant, hypotensive agent, antimutagenic agent, allergic inhibitor, cariostatic agent and deodorant - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a fruit polyphenol pressed and/or extracted from an unripe fruit of a fruit belonging to the family Rosaceae and comprising its purified fraction and an antioxidant, a hypotensive agent, an antimutagenic agent, an allergic inhibitor, a cariostatic agent and a deodorant comprising the polyphenol fraction pressed and/or extracted from the unripe fruit of the fruit belonging to the family Rosaceae and purified as an active ingredient. SOLUTION: The fruit polyphenol fraction having various kinds of physiological actions, especially antioxidizing properties, an angiotensin-converting enzyme(ACE) inhibitory activity, antimutagenic actions, a hyaluronidase inhibitory activity and a histamine liberation inhibitory activity, GTase activity inhibitory actions and deodorant and malodorous substance production inhibitory actions on malodorous substances can be obtained.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a fruit polyphenol, and more particularly to an antioxidant, a hypotensive agent, an anti-mutagenic agent, an allergy inhibitor, an anti-carious agent, and a deodorant containing a polyphenol fraction as an active ingredient. Agent.

[0002]

2. Description of the Related Art In the process of cultivating fruit trees such as edible fruits of the family Rosaceae, particularly apples, pears, and peaches, an operation called "cutting" is usually performed from mid-May to mid-July. The content of the work is to leave some fruits and discard others while the fruits that have set in a tuft after pollination are still immature, but this operation leaves a large amount of immature fruits unused. Has been disposed of. This immature fruit is extremely bitter compared to the mature fruit, and the cut surface of the fruit is easily browned. This fact suggests that polyphenol compounds are present in large amounts in immature fruits.

[0003] Polyphenol compounds are known as secondary metabolites of plants, which are ubiquitous in the plant kingdom, and are known to exist in abundant and large amounts. Some of these compounds have a long history of exhibiting a wide variety of physiological activities. It has attracted attention in the field of pharmacy and in recent years in the field of food chemistry.

[0004] Among them, particularly in tea polyphenols (catechins), which have been recently studied, antibacterial,
Very wide range of physiological effects, including anti-virus, anti-oxidation, anti-mutation, anti-cancer, suppression of platelet aggregation, suppression of blood pressure rise, suppression of blood glucose rise, reduction of blood cholesterol, anti-caries, anti-allergy, improvement of intestinal flora, deodorization Is being recognized. (JP-A-63-214183, JP-A-2-214)
6499, JP-A-4-178320, etc.)

[0005]

As described above, it is known that polyphenols extracted from tea, for example, have a wide range of physiological actions. In addition, a raw material search and production method for economically producing polyphenols, as well as the physiological effects of the polyphenols were examined and confirmed. As a result, they found that polyphenols having various physiological actions can be efficiently produced by using unripe fruits such as rose fruits, especially apples, pears, and peaches as raw materials, and by performing a specific treatment. did.

[0006]

That is, according to the present invention, apples, pears or peaches are extracted and / or extracted from unripe fruits and their fractions are purified. Include caffeic acid derivatives, p-coumaric acid derivatives, flavan-3-ols, flavonols, dihydrochalcones as polyphenol compounds, and condensed tannins which are regular polymers of catechins as high molecular polyphenol compounds. A polyphenol mixture is provided.

According to the present invention, the unripe fruits of apple, pear or peach are squeezed and / or extracted and purified, and their component compositions are as simple polyphenol compounds, caffeic acid derivatives and p-coumaric acid derivatives. , An antioxidant comprising, as an active ingredient, a polyphenol fraction containing flavan-3-ols, flavonols, dihydrochalcones, and condensed tannins which are regular polymers of catechins as high molecular polyphenol compounds. Is done.

Further, according to the present invention, the unripe fruit of apple, pear or peach is squeezed and / or extracted and purified, and its component composition is a simple polyphenolic compound such as a caffeic acid derivative or a p-coumaric acid derivative. ,
Effective is a polyphenol fraction having angiotensin I converting enzyme inhibitory activity, including flavan-3-ols, flavonols, dihydrochalcones, and condensed tannins which are regular polymers of catechins as high molecular polyphenol compounds. A blood pressure lowering agent is provided as a component. Furthermore, according to the present invention, it is obtained by squeezing and / or extracting and purifying immature fruits of apple, pear or peach, and its component composition is a simple polyphenol compound as a caffeic acid derivative, a p-coumaric acid derivative, or a flavan. -3-ols, flavonols, dihydrochalcone, and a polyphenol fraction having a mutagenicity-suppressing action, including a condensed tannin which is a regular polymer of catechins as a high-molecular-weight polyphenol compound, as an active ingredient An anti-mutagenic agent is provided.

Further, according to the present invention, the unripe fruit of apple, pear or peach is squeezed and / or extracted and purified, and its component composition is a simple polyphenolic compound such as a caffeic acid derivative or p-coumaric acid. Polyphenols having hyaluronidase inhibitory activity and histamine release inhibitory activity, including derivatives, flavan-3-ols, flavonols, dihydrochalcones, and condensed tannins which are regular polymers of catechins as high molecular polyphenol compounds An allergy inhibitor comprising a fraction as an active ingredient is provided. Furthermore, according to the present invention, it is obtained by extracting and / or extracting and purifying immature fruits of apples, pears or peaches, and its component composition is a simple polyphenol compound as a caffeic acid derivative, p-coumaric acid derivative or flavan. -3-ols, flavonols, dihydrochalcones, and polyphenol fractions having glucosyltransferase inhibitory activity, including condensed tannins, which are regular polymers of catechins as high molecular polyphenol compounds, are used as active ingredients. An anti-caries agent is provided.

[0010] Furthermore, according to the present invention, the unripe fruit of apple, pear or peach is squeezed and / or extracted and purified, and its component composition is a simple polyphenol compound as a caffeic acid derivative, p-coumaric. Deodorizing action on malodorous substances and production of malodorous substances, including acid derivatives, flavan-3-ols, flavonols, dihydrochalcones, and condensed tannins which are regular polymers of catechins as high molecular polyphenol compounds A deodorant comprising a polyphenol fraction having an inhibitory action as an active ingredient is provided.

[0011]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail.
The fruit polyphenol in the present invention, as described above,
Fruits belonging to the family Rosaceae, especially apples, pears, squeezed juice of immature fruits of peaches, consisting of polyphenol fraction purified from the extract, the purification of the polyphenol fraction, squeezed juice, extract It is carried out by treating with an adsorbent, and the fraction adsorbed by the adsorbent (hereinafter referred to as an adsorbed fraction) contains polyphenol. The polyphenol fraction is purified by eluting the adsorbed fraction with a hydrous alcohol (such as ethanol).

The polyphenol fraction is then subjected to a concentration treatment to obtain a liquid preparation. Furthermore, a powder preparation can also be obtained by spray-drying or freeze-drying the concentrated solution of the polyphenol fraction.

The raw materials used in the present invention include fruits belonging to the family Rosaceae, such as apples, pears and peaches, with apples being particularly preferred. As the fruits, immature fruits of apples, pears or peaches are used because they contain more polyphenol compounds and a large amount of various active ingredients having a wide range of physiological actions.

In the method of squeezing, the raw material is washed, and crushed and pressed as it is or with the addition of sulfurous acid to obtain a squeezed juice, and preferably a pectin-degrading enzyme is added. Next, there can be mentioned a method of obtaining a clear juice by means such as centrifugation and filtration. As an extraction method, the washed raw material is mixed with alcohol (ethanol, methanol, etc.), crushed, and immersed and squeezed, or extracted while heating and refluxing. Then, the alcohol is distilled off under reduced pressure, and then centrifuged. And a method of obtaining a clear extract by performing filtration and distribution or filtration with an organic solvent (hexane, chloroform, etc.).

As a purification method, an adsorbent capable of selectively adsorbing and eluting polyphenols, for example, a styrene-divinylbenzene synthetic adsorption resin, an anion exchange resin,
The polyphenol fraction is adsorbed by passing the above clear juice or clear extract through a column filled with octadecyl group chemically bonded silica gel (ODS) or the like. Then
After washing by passing distilled water, a 20 to 100% alcohol (eg, ethanol) solution, preferably about 50%
The polyphenol fraction can be eluted and recovered by passing the% alcohol solution through the column. The alcohol is distilled off by concentrating the obtained polyphenol solution under reduced pressure to obtain a fruit polyphenol liquid preparation (preferably, an organic acid such as malic acid is added). In addition, the liquid formulation as it is or a powder auxiliary such as dextrin is added,
By spray-drying or freeze-drying, a fruit polyphenol powder preparation can be obtained.

The composition of the fruit polyphenols obtained in the present invention includes simple polyphenol compounds such as caffeic acid derivatives, p-coumaric acid derivatives, flavan-3-ols (catechins), flavonols (quercetin glycosides), The present inventor has confirmed that a large portion is occupied by dihydrochalcones (phloretin glycosides) and the like, and condensed tannins and the like as high molecular polyphenol compounds.

Therefore, it is considered that the fruit polyphenol obtained in the present invention may have various physiological functions. As a result of the inventor's intensive studies, first, the polyphenol is a vegetable oil or fat. It has been found that a large amount of a component having an action of preventing the oxidation of linoleic acid is contained. That is, the fruit polyphenol obtained in the present invention is extremely effective as an antioxidant.

Second, it has been found that this polyphenol contains a large amount of a component having an effect of inhibiting the action of angiotensin I converting enzyme (hereinafter referred to as ACE), which is an enzyme involved in increasing blood pressure. Was. Therefore,
The fruit polyphenol obtained in the present invention is extremely effective as a blood pressure lowering agent.

The present inventor further studied and
When the ACE inhibitory substance in the fruit polyphenol was searched for, it was confirmed that it was a condensed tannin represented by the structural formula in FIG. In addition, many recent research results show that carcinogenic substances and mutagenic substances have a high correlation, indicating that carcinogenicity and mutagenicity are closely related. That is, a substance that suppresses mutagenicity can be expected to prevent carcinogenicity. Then, it was confirmed whether or not the fruit polyphenol obtained by the present invention has an antimutagenic effect, and it was found that the effect was effective. Therefore, the fruit polyphenol obtained in the present invention is extremely effective also as an anti-mutagenic agent.

Furthermore, histamine is known as an in vivo chemical mediator causing various allergic symptoms, and is released mainly from mast cells and basophils by degranulation. Hyaluronidase is an enzyme that is thought to form an integral part of cellular activity in connective tissue remodeling, but has been studied recently (Chem. Pharm. Bu).
ll. 33. p642.1985, ibid. 33.198
5, ibid. 33. p3787.1985, ibid. 33. p5
079.1985, ibid. 40.1439.1992), the activity of inhibiting hyaluronidase and the activity of inhibiting histamine release from mast cells show a high correlation with synthetic anti-allergic agents (sodium cromoglycate and tranilast). Several anti-allergic substances have been searched for in the products. Then, it was confirmed whether or not the fruit polyphenol obtained by the present invention has a hyaluronidase inhibitory activity and a histamine release inhibitory activity, and it was found that the fruit polyphenol had the action and effect. Therefore, the fruit polyphenol obtained in the present invention is extremely effective as an allergy inhibitor.

Furthermore, today, caries (cavities) are St.
Although it has been confirmed that it is a kind of bacterial infection caused by oral streptococci, mainly reptococcus mutans, plaque formation is considered to be a particularly important factor in the process of caries development. That is, sucrose contained in food is converted to glucosyltransferase (hereinafter referred to as GT)
As a result, adhesive insoluble glucan is synthesized, via which bacteria adhere to the tooth surface, and the resulting plaque causes dental caries.

From this, it is expected that a substance that inhibits GTase activity can be applied as an effective anti-cariogenic agent (Appl. Env. Microbiol., 59 (4), 968-973, 1993;
Biosci. Biotech. Biochem., 56 (5), 766-768, 1992, Agric.
Biol.Chem., 54 (11), 2925-2929,1990, Chem.Pharm.Bul
l., 38 (3), 717-720, 1990). Therefore, in the present invention,
S. sobrinus, one of the representative species of cariogenic bacteria, produces
When the ability of fruit polyphenols to inhibit insoluble glucan-producing GTase was examined, it was found that they had an effect. Therefore, the fruit polyphenol obtained in the present invention is extremely effective also as an anti-cariogenic agent.

Further, among the malodorous ingredients derived from foods,
It is said that volatile sulfur compounds, such as hydrogen sulfide, methyl mercaptan, and dimethyl sulfide, are proportional to the intensity of bad breath and the content (Nippon Periodon, 17,233-245, 1975). Among them, a strong correlation with methyl mercaptan concentration has been observed (Journal of Periodontology, 18, 1-12, 1976 and Bulletin of the Dental Society, 76, 1
923,1976). Then, when the deodorizing effect of the fruit polyphenol obtained in the present invention on methyl mercaptan and dimethyl sulfide was measured, it was found that the effect was effective. Furthermore, the effect of inhibiting the production of fruit polyphenols on methyl mercaptan, dimethyl sulfide and dimethyl disulfide was measured, and it was found that the effects were also obtained. Therefore, the fruit polyphenol obtained in the present invention is extremely effective as a deodorant.

[0024]

Next, the present invention will be described in more detail with reference to examples, but the present invention is not limited to these examples.

(Example 1) [Component analysis of immature apple juice] The following samples were analyzed for general components. -Sample treatment method Fruit samples were crushed and squeezed with a mixer while adding an appropriate amount of potassium metabisulfite as an antioxidant. The squeezed liquid was subjected to the following measurement as clear juice by centrifugation and filtration.

Measurement items (and methods) Average weight per individual (n = 50) Squeezing rate (%) pH Acidity (g / l in terms of malic acid) Brix Total phenol (in terms of chlorogenic acid, ppm -Total ascorbic acid-Organic acid composition-Sugar composition-Metal ion composition-Free amino acid composition The measurement results are shown in Table 1.

[0027]

[Table 1]

As can be seen from Table 1, immature apple juice showed a large difference in components as compared with mature juice. It should be emphasized that it is rich in phenol components, ascorbic acid, organic acids (particularly quinic acid), metal ions, and free amino acids (particularly asparagine, phenylalanine, and γ-aminobutyric acid). On the other hand, saccharides were very low.

Next, the composition of the polyphenol component of the immature apple was examined by high performance liquid chromatography.
As a measurement sample, a polyphenol component purified from a fruit extract by a solid phase extraction method (see Example 2) was used. FIG. 1 shows the measurement results.

From the above, the simple polyphenol compound composition of immature apple fruit is as follows: caffeic acid derivatives (chlorogenic acid, etc.), p-coumaric acid derivatives, flavan-3-ols (catechin, epicatechin, etc.), flavonols (quercetin) Glycosides) and dihydrochalcone (phlorethin glycosides, especially phlorizin) were found to account for the majority. Quantitatively, chlorogenic acid, catechin, epicatechin,
The phlorizin content was assumed to be high.

(Example 2) [Antioxidant action of immature fruit polyphenol]-Raw materials The following fruits were used as raw materials. Apple mature fruit: commercial product (without bag cultivation) Apple immature fruit: collected in mid-June "Fuji"-average weight 4.97 g (n = 50) "Tsugaru"-average weight 7.80 g (n = 50) ) "Jonagold"-average weight 3.86 g (n = 50) "Hokuto"-average weight 3.32 g (n = 50) "Oh Lin"-average weight 10.34 g (n = 50)

Pear immature fruit: collected in early June "Hosui"-average weight 8.98 g (n = 50) Peach immature fruit: collected in early June. "AKATSUKI"-average weight 5.03 g (n = 50)

Sample Preparation Fruit Juice A clarified juice was obtained in the same manner as in Example 1. Extracted sample-As an extraction method, 400 g of the raw material is 1%
After homogenizing with hydrochloric acid-acidic methanol, the mixture was extracted with heating under reflux (3 times). The extract was concentrated under reduced pressure to remove methanol, and then added with chloroform and partitioned (2).
Times), the aqueous layer was collected, filtered, and made up to 200 ml with distilled water to obtain an extracted sample. In addition, if necessary, a polyphenol component was purified from a fruit juice sample and an extracted sample by a solid phase extraction method using Sep-pak C18.

-Antioxidant test method The antioxidant test method was as follows. That is, the substrate solution (4
% Linoleic acid in 5 ml of phosphate buffer (p
H7.0) After mixing 4 ml of the test solution and 1 ml of the test solution in a sealed test tube, the mixture was stored in a thermostat at 50 ° C. protected from light. At this time, a sample to which ethanol was added instead of the substrate as a control, and a sample to which a sample solvent was added instead of the sample as a blank were stored under the same conditions.

During the storage period, the reaction solution was sampled with time, and the produced peroxide was quantified by the isothiocyanate method (rodane iron method).

-Measurement Results The immature and mature extracts of apples (Fuji) were examined for antioxidant activity. When “Fuji” extract of immature and mature fruits was added to the reaction system in the range of 1-1000 μl / gLA, 1
FIG. 2 shows a graph of the amount of lipid peroxide formed (absorbance value at 500 nm) and the concentration added after a week. From now on, "Fuji"
Antioxidant activity against linoleic acid was observed in both the immature and mature extracts, but a strong effect was particularly observed in the immature extract.

Subsequently, the extract of immature and mature fruits was extracted with Se
It was roughly fractionated into two by p-pak C18 (fraction A:
Sep-pak C18 non-adsorbed fraction. Fraction B: Sep-pak C18 adsorption fraction (polyphenol fraction). Extract and each fraction
1 When added to the reaction system in the range of -1000 μl / gLA, 1
FIG. 3 is a graph showing the relationship between the amount of lipid peroxide produced (absorbance 500 nm value) and the concentration added after a week.

As a result, it was found that the antioxidant property of the mature fruit extract was derived from the component of fraction B. In the immature fruit extract, fractions A and B exhibited antioxidant activity, but fraction B exhibited a stronger effect. The above results strongly suggested that the antioxidant properties observed in both extracts were mainly attributable to the presence of the component of Fraction B, ie, the polyphenol compound.

Further, among the simple polyphenol compounds contained in apples, those which can be obtained in pure form were examined for their antioxidant properties. FIG. 4 shows a comparison graph of the amount of generated lipid peroxide (absorbance 500 nm value) after one week when 2 μmol / gLA of each substance was added to the reaction system. In FIG. 4, in addition to the polyphenol compound, three kinds of known antioxidants BHA, BHT, and vitamin E were used as comparative substances, and quinic acid, malic acid, γ were used as typical components in the fraction A.
-Aminobutyric acid (GABA) was also studied.

As a result, among the polyphenol compounds, 6 of caffeic acid, chlorogenic acid, (+)-catechin, (-)-epicatechin, quercetin and rutin (quercetin-3-glu-rha) were used.
The species had antioxidant power comparable to BHA and BHT.
However, even with the same polyphenol compound, p-coumaric acid, phloretin, and phlorizin did not show any antioxidant action. Also, none of the components in fraction A had an antioxidant effect. From the comparison between the content of each component and the antioxidant power in apples, the strong antioxidant action exerted by the extract and juice of apple immature fruits is mainly due to chlorogenic acid, (+)-catechin, (-)-epi It was presumed to be due to the presence of catechin.

From the results thus far, it has been clarified that immature fruits of apple (Fuji) have a strong antioxidant activity and their active ingredients. Subsequently, it was examined whether immature fruits of apples other than "Fuji" also have the same effect. Various immature apples (Fuji, Tsugaru, Jonagold)
For fraction B (polyphenol fraction) of the extract of “Hokuto” and “Wang Lin”, when each fraction was added to the reaction system in the range of 1-500 μl / gLA, the peroxidation produced after one week FIG. 5 shows a graph of the lipid amount (absorbance 500 nm value) and the added concentration.

FIG. 5 also shows the results of mature fruits of "Fuji" as a control. As a result, strong antioxidant properties were observed in all apple varieties, and almost no difference in antioxidant power was observed between the varieties.

Further, the antioxidant effect of immature fruits other than apples was also examined. For fraction B (polyphenol fraction) of the extract of various fruits immature fruits (apple “Fuji”, pear “Hosui”, peach “Akatsuki”), each fraction was 1-500
FIG. 6 shows a graph of the amount of lipid peroxide produced (absorbance 500 nm value) and the concentration added one week after addition to the reaction system in the range of μl / gLA.

As a result, it was found that immature peaches had almost the same antioxidant power as immature apples, and immature pears had a very strong antioxidant power, although not as strong as apples.

(Example 3) [Inhibition of angiotensin I converting enzyme (ACE) by unripe fruit polyphenols]-Raw materials The following fruits were used as raw materials. Immature apple: Same as in Example 2-Sample preparation: An "extracted sample" and a "polyphenol fraction" were prepared in the same manner as in Example 2.

ACE Inhibition Test Method The ACE inhibition test was carried out according to a conventional method. That is, commercial A
A sample solution was added to the CE solution, and after pre-incubation, Bz-Gly-His-Leu was added as a substrate and reacted. After fluorescent labeling the His fragment generated by the reaction with orthophthaldialdehyde, the fluorescence intensity (Ex. 360 nm, Em. 490 nm)
Was measured.

At this time, the fluorescence intensity of the test solution was S, the value when water was added instead of the sample was C, and the value of the enzyme blank for S (water was added instead of the enzyme) was S _B and C. the value of the enzyme blank as C _{B of, {1- (S-S B} ) / (CC B)} × 100
The ACE inhibitory activity was expressed as (%).

-Measurement results- Extracts of immature apples and mature fruits (both "Fuji") were separated from the non-adsorbed fraction (→ fraction A) and polyphenol fraction (→ fraction) using Sep-pak C18. Fraction B). The ACE inhibitory activity of each fraction was examined. The results are shown in FIG. As a result, in both immature and mature fruits, fraction A was strong in fraction B.
CE inhibitory activity was observed. Especially in the fraction B of immature fruit, 10
Although 0% inhibition was observed, in order to compare the strengths of both inhibitory activities in more detail, the change in the ACE inhibitory activity when the amount of both fractions B added to the reaction system was changed was examined. Was. The results are shown in FIG.

As a result, the 50% inhibitory concentration (IC ₅₀ ) of the fraction B of the mature fruit against ACE was about 3 ul, whereas the fraction B of the immature fruit had a very strong activity of 0.1 ul or less. Indicated. This strong inhibitory activity in immature fruits was similarly observed in cultivars other than "Fuji". From the above results, it was strongly suggested that immature apple fruits have a strong inhibitory substance against ACE and that the active substance is a polyphenol compound. ACE of pear and peach immature fruit
The inhibitory activity was weaker than that of apple.

Next, a simple polyphenol compound which is found in immature fruits of apples and which can be obtained as a pure product is as follows:
The presence or absence of ACE inhibitory activity was examined. Specifically, the variation in ACE inhibitory activity when the concentration of each substance added to the reaction system was changed was examined so that the intensities of the activities between the substances could be compared. The results are shown in FIG.

In FIG. 9, for comparison, catechins of tea already known to have ACE inhibitory activity were also measured. As a result, the IC ₅₀ values of (−)-epigallocatechin gallate (EGCg) and (−)-epicatechin gallate (ECg) were 0.3 mM and 2 mM, respectively, indicating strong inhibitory activity. In addition, there is a report that "GABARON tea", which has increased GABA concentration in tea leaves by processing, showed a blood pressure lowering effect in an experiment using spontaneously hypertensive rats.
Showed no ACE inhibitory activity.

Although the important apple phenols had inhibitory activity at high concentrations, the IC ₅₀ values were low.
Even the most active quercetin is about 5 mM, and EGCg
The inhibitory activity was much weaker than that of ECg and ECg.

Here, assuming that all the polyphenols in the apple immature fruit fraction B are EGCg, the total phenol amount in the fraction B was calculated from the calibration curve of EGCg, and it was found to be about 15,000 ppm.
And the polyphenol concentration of fraction B was 33.8% in terms of EGCg.
It was calculated to be equivalent to 2 mM. Using this calculated value and plotting the activity curve of fraction B on FIG. 9, a strong inhibitory activity exceeding EGCg (IC ₅₀ = 0.2 mM) was observed. In practice, EGCg and ECg are not present in fraction B, and no remarkable inhibitory activity is observed with other simple polyphenols as described above. It was strongly suggested that the strong ACE inhibitory activity of the immature fraction B observed in FIG. 9 was due to the presence of other unidentified polyphenols present in the mixture.

(Example 4) [Identification of ACE Inhibitor in Polyphenol of Unripe Apple Fruit]-Sample As in Example 3, an extract sample was obtained from unripe apple "Fuji", and polyphenol was extracted by solid phase extraction. The fractions were purified to give the following samples. -Experimental method The sample solution was loaded on a Sephadex LH-20 column. After washing the column with distilled water, polyphenol was eluted and fractionated using 20% to 60% methanol (acidic with hydrochloric acid) and 70% acetone (acidic with hydrochloric acid) in order. About the obtained fraction,
HPLC composition analysis, total phenol determination, and ACE inhibition test were performed. Further, absorption spectrum measurement and gel permeation chromatography were performed as necessary.

-Experimental results Sephadex LH-20 colum was used as a specific means for isolating the ACE inhibitory activity in the sample from other polyphenols.
Tried fractionation by n. The eluted fractions obtained by the experiment were subjected to simple polyphenol analysis by HPLC and an ACE inhibition test. As a result, it was confirmed that all the simple polyphenols were completely eluted when 60% methanol was eluted.

The ACE inhibitory activity was determined by the following elution solvent, 70
% Acetone eluted fraction. The isolated fraction is
After the solvent was distilled off, the residue was freeze-dried and powdered. The active substance was readily soluble in water (the aqueous solution was orange) and about 0.7 g was obtained from 100 ml of fraction B. Next, the obtained powder was dissolved in water, and the absorption spectrum was measured with a spectrophotometer. Figure 1 shows the results
0.

The obtained spectrum showed a maximum absorption at 280 nm, and its shape was very similar to that of (+)-catechin or (-)-epicatechin. When the aqueous solution was autoclaved at 120 ° C. for 10 minutes, the solution turned red, and it was considered that anthocyanidins were formed. These results strongly suggested that the active form was procyanidins (regular polymers of catechins) known as condensed tannins. However etc. elution behavior in dissolution behavior as well as silica gel HPTLC in the previous Sephadex LH-20, this substance is not a dimer of about polymerization degree such as procyanidin B _2, is estimated to be a higher order polymeric material Was.

Therefore, the molecular weight of this substance was measured by GPC. In general, polyphenol compounds have strong affinity for column packing due to hydrophobic bonds or hydrogen bonds, and therefore GFC (Gel Filtrati
on Chromatography) is considered difficult.
Therefore, after acetylating the phenolic hydroxyl group of the active substance with pyridine / acetic anhydride to make it soluble in organic solvents, GPC analysis in organic solvents (THF) was attempted. The results are shown in FIG.

As a result, a single broad peak appeared on the chromatogram. At the same time, the average molecular weight of the substance was calculated to be about 2,000 from a molecular weight calibration curve using polystyrene. Although not shown here, the structural formula of the ACE inhibitory active substance estimated at the present time is shown in FIG. 12 after considering the results of other experiments (partial decomposition with toluene-α-thiol, FAB-MS measurement, etc.). Was. From this, it was determined that this substance was a kind of condensed tannin.

The purified polyphenol fraction was separated from Sephadex
When fractionating with LH-20, the total phenol amount was measured before fractionation (that is, the purified polyphenol fraction) and after fractionation (70% acetone-eluting fraction: that is, the condensed tannin fraction), and the values were compared. did. The results are shown in Table 2.

[0061]

[Table 2]

From this, it was found that about half of the total polyphenols in the immature apple fruit was the condensed tannin, which was the most quantitative component. The ACE of this substance
It was calculated IC ₅₀ for, IC ₅₀ of EGCg by weight Comparative
, Which is a very low value of about 1/10 or less, that is, it was confirmed that this substance is a very powerful ACE inhibitory substance. From the above results, it was determined that apple immature fruit polyphenol could be a powerful ACE inhibitor.

[0063] (Example 5) was crushed with [unripe fruit Preparation of polyphenol] apple unripe fruits (5 to 10 g / piece) adding an appropriate amount while crusher SO ₂ to about 50 kg, were pressed in an oil press. About 50 ppm of pectin-degrading enzyme was added to the obtained fruit juice, subjected to centrifugation or diatomaceous earth filtration, and further subjected to microfiltration to obtain 35 L of clear juice. Next, the clarified juice was passed through a column filled with a styrene-divinylbenzene-based industrial synthetic adsorption resin (6 L). Furthermore, 0.
After removing sugars by passing through 6 L of 1% HCl-containing water, 0.1 mL of water was added.
Elution with 50% ethanol containing 1% HCl yielded 3 L of the major polyphenol fraction.

The obtained fraction was concentrated under reduced pressure with an evaporator to a concentrated fraction of 1.5 L, and dried with a spray dryer.
228.2 g of an unripe apple fruit polyphenol powder preparation was obtained. The data on the recovery are described below.

Column recovery rate: 95.6% Spray drying recovery rate: 93.0% Recovery rate from fruit juice; 0.65% Powder recovery rate from polyphenol in fruit juice; 88.9%

(Example 6) [Anti-mutagenic effect of immature fruit polyphenol] In this example, the anti-mutagenicity was measured by the Ames method (Muta).
Tion Research, vol. 31, p34
7, 1975). -Same as raw materials and sample preparation Examples 2, 3 and 4. However, the “condensed tannin” obtained in Example 4 is hereinafter abbreviated as “apple tannin”.

-Measuring method of antimutagenicity Mutagenic substances (benzo (a) pyrene, Trp-P-2)
Phosphate buffer, sample solution, S9-mix, Salmonella typhimurium TA98 or TA
100) After mixing the overnight culture and culturing at 37 ° C. for 2 days,
The number of colonies that appeared was counted. Each polyphenol had an exponential concentration gradient between 1 and 300 (μg / plate), and the mutagenic activity was calculated by the following equation.

Mutagenic suppression activity rate = ｛(CB) − (S
−B)｝ / (CB) × 100 (%)

At this time, the number of colonies of the test solution was S, the control to which water was added instead of the sample was C, and the blank to which water was added instead of the sample and the mutagen was B.

-Results For various polyphenols and apple tannins, Trp
FIG. 13 shows the mutagenic inhibitory activity rate for -P-2, and FIG. 14 shows the mutagenic inhibitory activity rate for benzo (a) pyrene. The vertical axis represents the mutagenicity-suppressing activity rate, which is 100% when the activity is suppressed to the same level as spontaneous reversion (blank). The horizontal axis shows the sample amount per plate.

As a result, apple tannin inhibited the mutagenicity of each mutagen in a concentration-dependent manner, and exhibited an inhibitory effect at a concentration lower than or equal to that of epigallocatechin gallate. Under the conditions of this experiment, about 17 μg of apple tannin (about 51 μg of epigallocatechin gallate) per 1 μg of Trp-P-2, and about 37 μg (about 56 μg of the same) for 5 μg of benzo (a) pyrene; % Mutagenic activity. In other words, this polyphenol
When applied to a mutagenic substance (carcinogenic substance), a large amount of a component that strongly suppressed the mutagenicity was contained. Therefore, the fruit polyphenols obtained in the present invention are extremely effective as anti-mutagenic agents.

(Example 7) [Hyaluronidase inhibitory action and histamine release inhibitory action of fruit immature fruit polyphenol] In this example, the allergic inhibitory action is the hyaluronidase inhibitory activity and the histamine release inhibitory action using animal cells. It was measured as an index. -Raw material apple immature fruit, pear immature fruit, peach immature fruit, apple maturity fruit-Sample preparation Each fruit extract was prepared in the same manner as in Example 2, and the "polyphenol fraction" was further separated using Sephadex LH-20. The purified product was used as apple tannin.

-Method for measuring hyaluronidase inhibitory activity The method for measuring hyaluronidase inhibitory activity is basically described in J. Org. Bio
l. , Vol. 250, p79, modified by 1975. That is, a sample solution was added to a commercially available hyaluronidase solution, and after preincubation, a 48/80 compound solution of a histamine releasing agent was added, hyaluronidase was activated at 37 ° C., and a hyaluronic acid solution was added as a substrate and reacted. The N-acetylglucosamine produced by the reaction was colored by the Morgan-Elson method, measured by absorbance (586 nm), and the hyaluronidase inhibitory activity was calculated by the following equation.

Hyaluronidase inhibitory activity rate = ｛(CC)
_B ) − (S−S _B )｝ / (C−C _B ) × 100 (%)

[0075] At this time, the absorbance of the test solution S, the plus buffer instead of the sample C, the enzyme blank S _B of S, the enzyme blank C was C _B.

Assay for Histamine Release Inhibitory Activity The method for measuring histamine release inhibitory activity is described in J. Food Hyg. Soc. Japa
n, vol. 35, p497, 1994. That is, rat basophil leukemia cells RBL-2H3 were transformed with 10% FBS, α-
After culturing in MEM medium (37 ° C., 5% CO ₂ ), antibody DN
P-lgE was allowed to act for 90 minutes, and HEPES buff
After washing with er, the antigen DNP-BSA is allowed to act for 35 minutes. Apple components are added during the action of the antibody or antigen. To stop the reaction, an ice-cold HEPES buffer was added, the extracellular solution was taken, perchloric acid was added, and after cooling, centrifugation was performed. Then, 0.45 µ filtrate was analyzed by HPLC to determine the amount of histamine released from the cells. It was measured. The histamine release inhibitory activity was calculated by the following equation.

Histamine release inhibitory activity rate = ｛(CB)
− (S−B)｝ / (C−B) × 100 (%)

At this time, the amount of histamine in the test solution was S, the sample to which HEPES buffer was added instead of the sample was C, and the blank to which no antibody or antigen was added was B.

-Results The hyaluronidase inhibitory activity ratio of each fruit extract, sodium cromoglycate and apple tannin is shown in FIG.
And FIG. As a result, each fruit extract and apple tannin inhibited hyaluronidase in a concentration-dependent manner, and their activities were observed. When compared with the synthetic anti-allergic agent sodium cromoglycate as a control, 50
As for the IC50 value which is a concentration that inhibits the% activity, sodium cromoglycate was about 0.051 mg / ml, while apple tannin was a value close to about 0.086 mg / ml.

When the antigen DNP-BSA for each fruit extract and apple tannin acts, the antibody DNP-lgE
17 and FIG. 1 show the histamine release inhibitory activity rate during the action.
8 is shown. As is apparent from these results, apple tannin contained in the polyphenol fraction was found to have a concentration-dependent histamine release inhibitory activity. From the above, the polyphenol contained a large amount of a component having an action of inhibiting the action of hyaluronidase, which is an enzyme relating to type I allergy, and a component having an action of inhibiting histamine release. Therefore, the fruit polyphenol obtained in the present invention is extremely effective also as an allergy inhibitor.

(Example 8) [Glucosyltransferase inhibitory action of immature fruit polyphenol] In this example, the effect of fruit polyphenol on the insoluble glucan-producing GTase produced by S. sobrinus, one of the representative species of cariogenic bacteria, was examined. The inhibitory ability was examined. -Raw material and sample preparation Same as in Examples 6 and 7-Bacteria used; Streptococcus sobrinus ATCC 334
78

Preparation of GTase; GTa of S. sobrinus
se was prepared by the following method. S. sobrinus was transformed into a TTY medium (Agric. Biol. Chem., 54 (11), 29
25-2929, 1990) at 37 ° C. for 18 hours, and the cells were removed by centrifugation to obtain a supernatant. After ammonium sulfate was added to the supernatant to 50% saturation, the precipitate formed by centrifugation was collected. The precipitate was prepared in 0.05M phosphate buffer (pH
After redissolving in 6.5), the mixture was dialyzed against the same buffer. GTase in the dialysate was purified and recovered by hydroxylapatite chromatography. At this time, insoluble glucan-producing GTase was eluted at a phosphate buffer concentration of about 0.4 M. The eluted fraction was collected and subjected to the following experiment.

-Gase Inhibitory Activity Assay A purified GTase solution and a sample were placed in 1 ml of a substrate solution (0.1 M phosphate buffer (pH 6.5) containing 2% sucrose, 0.1% sodium azide, and 40 μM dextran T10). The solution was added and diluted to a total volume of 2 ml with water.
The reaction was performed for 8 hours. The amount of insoluble glucan produced by the reaction was measured as turbidity indicated by absorbance at 550 nm, and the production rate was calculated from the following equation. Insoluble glucan generation rate = (SS-SB) / (CS-C
B) × 100 (%) SS: sample SB: sample enzyme blank CS: control CB: control enzyme blank

-Results The polyphenol fraction of each fruit extract and the GTase inhibitory activity of each polyphenol component are shown in FIGS. The vertical axis represents the insoluble glucan production rate, which was calculated assuming that the production amount in the control with no sample added was 100%. The horizontal axis indicates the amount of the sample or polyphenol added to the reaction system by volume or concentration. As shown in FIG. 19, among the polyphenol fractions of the various fruit extracts, a remarkable GTase inhibitory activity was observed particularly in the polyphenol fractions of “Fuji” immature fruits and “Shintaka” immature fruits. At this time, the 50% inhibition amount of the matured “Fuji” fruit against GTase was about 50%.
μl, whereas that of unripe "Fuji" is about 0.7
From μl, it was found that immature apple fruits had about 70 times stronger GTase inhibitory activity than mature fruits.

Next, the GTase inhibitory activity of various polyphenol components was examined mainly on the components contained in apple. As shown in FIG. 20, among the components in apple, the chlorogenic acid and phlorizin had very weak GTase inhibitory activities. On the other hand, among the various catechins, the inhibitory activity of the epicatechin monomer was weak, whereas the high molecular epicatechin polymer represented by the deduced chemical structural formula in FIG. 12 (indicated as apple tannin in FIG. 20) was strong. GT
It was found to have ase inhibitory activity. In FIG. 20, as a control, epicatechin gallate (Agric. Biol. Chem., 5 (11), 2925-2929, 1990, Ch.) Which is a representative catechin of green tea already known to be a GTase inhibitor.
The results of em. Pharm. Bull., 38 (39, 717-720, 1990) are shown at the same time.
While the 0% inhibitory concentration was about 200 ppm, that of apple tannin was calculated to be about 2 ppm, which is about 100 times as potent. From these results, it was estimated that the strong GTase inhibitory activity of the immature fruits of "Fuji" shown in FIG. 19 was due to the presence of this substance. based on the above results,
The fruit polyphenol obtained in the present invention is a powerful GTas
Since it has an e-inhibitory active ingredient, it is considered to be extremely effective as an anti-cariogenic agent.

(Example 9) [Deodorizing effect of unripe fruit polyphenol] The deodorizing effect test in this example was carried out by the method of Ui et al. (Nisshoku Kogyo, 38,109).
8-1102, 1991). -Sample A spray-dried polyphenol fraction (apple polyphenol powder preparation) purified from the unripe apple fruit juice described in Example 2 was used. Further, as a control, a commercially available green tea polyphenol preparation and sodium copper chlorophyllin, which are known deodorants, were used.

Test Method for Deodorizing Effect 1 ppm methyl mercaptan solution and 1 ppm
A ppm dimethyl sulfide solution was prepared. 1 ml of a 0.1 M phosphate buffer (pH 7.5) containing each concentration of a sample was placed in a vial having a content of 30 ml (for comparison, no sample was added), and 1 ml of a 1 ppm methyl mercaptan solution was added thereto.
Alternatively, 1 ml of a 1 ppm dimethyl sulfide solution was added, immediately stoppered and stirred. After incubating this at 37 ° C. for 5 minutes, 5 ml of head space gas was injected into the GC,
The peak height of methyl mercaptan or dimethyl sulfide was measured.

-Measurement Results Each sample was examined for its deodorizing effect on malodorous components. 0.05-0.5% or 0.05-5% of sample
21 and 22 show the relationship between the concentration of methyl mercaptan or dimethyl sulfide in the headspace gas and the concentration of the sample added when added to the reaction system in the range of. As is clear from these results, it was confirmed that the apple polyphenol powder preparation had a deodorizing effect on methyl mercaptan and dimethyl sulfide in a concentration-dependent manner.

(Example 10) [Effect of immature fruit polyphenol on the production of malodorous substances]
The test for inhibiting the production of offensive odors in this example was performed by a modified method of the method of Ui et al. (Nisshoku Kogyo, 38, 1098-1102, 1991). -Sample As in Example 9, an apple polyphenol powder preparation, a commercially available green tea polyphenol preparation and copper chlorophyllin sodium were used.

Test Method for Inhibitory Effect on Production of Odorous Substances Saliva spontaneously discharged was collected from persons who had prohibited eating and drinking for 2 hours before collecting saliva. A sample containing each concentration and 40 mM L-methionine in a vial having a content volume of 30 ml.
1 ml of a 1 M phosphate buffer (pH 7.5) was added (a control was added without a sample), and 1 ml of saliva collected in advance was added thereto, and the mixture was immediately stoppered and stirred. After incubating this at 37 ° C. for 24 hours, 5 ml of headspace gas was added to G
C, methyl mercaptan produced by the reaction,
The peak heights of dimethyl sulfide and dimethyl disulfide were measured.

-Measurement results Each sample was examined for its effect on suppressing the production of malodorous substances.
Methyl mercaptan in the headspace gas when the sample was added to the reaction system in the range of 0.0015-0.5%,
The relationship between the concentrations of dimethyl sulfide and dimethyl disulfide and the sample addition concentration is shown in FIGS. As is evident from the results, it was confirmed that the apple polyphenol powder preparation had a concentration-inhibiting effect on the production of methyl mercaptan, dimethyl sulfide and dimethyl disulfide in a concentration-dependent manner.

[0092]

As described above, according to the present invention,
Since immature fruits of apples, pears or peaches are used as raw materials and subjected to a specific treatment, fruit polyphenols having various physiological actions, in particular, antioxidant properties and ACE inhibitory activity can be provided. Further, according to the present invention, an antioxidant containing a polyphenol fraction having an antioxidant effect as an active ingredient can be provided.

Further, according to the present invention, there can be provided a hypotensive agent comprising a polyphenol fraction having an angiotensin I converting enzyme inhibitory activity as an active ingredient. Further, according to the present invention, it is possible to provide an anti-mutagenic agent comprising a polyphenol fraction having a mutagenic inhibitory effect as an active ingredient. Furthermore, according to the present invention, there can be provided an allergy inhibitor comprising a polyphenol fraction having a hyaluronidase inhibitory activity and a histamine release inhibitory activity as an active ingredient.

Further, according to the present invention, it is possible to provide an anti-cariogenic agent comprising a polyphenol fraction having glucosyltransferase inhibitory activity as an active ingredient. Furthermore, according to the present invention, it is possible to provide a deodorant containing a polyphenol fraction having an effect of deodorizing malodorous substances and inhibiting production of malodorous substances as an active ingredient.

[Brief description of the drawings]

FIG. 1 is a graph showing a comparison between a wavelength-variable chromatogram of an immature apple polyphenol and an ultraviolet absorption spectrum of each peak.

FIG. 2: The amount of lipid peroxide produced (absorbance 500 nm) one week after apple “Fuji” extract was added to the reaction system
2) is a graph showing the relationship between the value and the additive concentration.

FIG. 3 shows the amount of lipid peroxide produced (absorbance 500 nm) after one week when each fraction of apple “Fuji” was added to the reaction system.
2) is a graph showing the relationship between the value and the additive concentration.

FIG. 4 shows the amount of lipid peroxide produced (absorbance 500 nm) one week after the addition of the polyphenol compound to the reaction system.
Value).

FIG. 5 shows the amount of lipid peroxide produced (absorbance) one week after each apple polyphenol fraction was added to the reaction system.
500 nm value) and a graph showing the relationship between the addition concentration.

FIG. 6 shows the amount of generated lipid peroxide (absorbance) after one week when each immature fruit polyphenol fraction was added to the reaction system.
500 nm value) and a graph showing the relationship between the addition concentration.

FIG. 7 is a graph showing the ACE inhibitory activity of each fraction.

FIG. 8 is a graph showing ACE inhibitory activity when the amount of Fraction B added was changed.

FIG. 9 is a graph showing ACE inhibitory activity when the concentration of each substance added to the reaction system was changed.

FIG. 10 is a graph showing an absorption spectrum of an isolated fraction measured by a spectrophotometer.

FIG. 11 is a graph showing the results of measuring the molecular weight by GPC.

FIG. 12 is a structural formula of a putative ACE inhibitory active substance.

FIG. 13 is a graph showing the mutagenicity-suppressing activity of polyphenols against Trp-P-2.

FIG. 14 is a graph showing the mutagenicity inhibitory activity of polyphenols on benzo (a) pyrene.

FIG. 15 is a graph showing the hyaluronidase inhibitory activity of various fruit extracts.

FIG. 16 is a graph showing the hyaluronidase inhibitory activities of an antiallergic agent and apple tannin.

FIG. 17 is a graph showing the histamine release inhibitory activity ratio of each fruit extract and apple tannin when the antigen DNP-BSA is acting and when the antibody DNP-lgE is acting.

FIG. 18. Antigen DNP-BSA for apple tannin
The graph which shows the histamine release inhibitory activity rate at the time of action | operation, and the antibody DNP-lgE action.

FIG. 19 is a graph showing the relationship between the amount of insoluble glucan produced and the amount added when each fruit polyphenol fraction was added to the reaction system.

FIG. 20 is a graph showing the relationship between the amount of insoluble glucan produced and the concentration added when each polyphenol compound was added to the reaction system.

FIG. 21 is a graph showing the residual amount of methyl mercaptan when apple polyphenol, green tea polyphenol, and sodium copper chlorophyllin were added to the reaction system.

FIG. 22 is a graph showing the residual amount of dimethyl sulfide when apple polyphenol, green tea polyphenol, and sodium copper chlorophyllin were added to the reaction system.

FIG. 23 is a graph showing the amount of methyl mercaptan generated when apple polyphenol, green tea polyphenol, and sodium copper chlorophyllin were added to the reaction system.

FIG. 24 is a graph showing the amount of dimethyl sulfide generated when apple polyphenol, green tea polyphenol, and sodium copper chlorophyllin were added to the reaction system.

FIG. 25 is a graph showing the amount of dimethyl disulfide generated when apple polyphenol, green tea polyphenol, and sodium copper chlorophyllin were added to the reaction system.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI theme coat ゛ (Reference) A61P 1/00 A61P 1/00 1/02 1/02 9/12 9/12 37/08 37/08 43 / 00 111 43/00 111 // C07D 311/30 C07D 311/30 311/62 311/62 (72) Inventor Shunji Shimoda 4-1-5 Higashinaka Shinjuku, Kashiwa City, Chiba Pref. 4C062 EE49 FF44 4C083 AA111 AA112 AC211 AC212 AC841 AC842 CC41 EE18 EE34 4C086 AA01 AA02 BA08 MA03 MA04 NA14 ZA42 ZA66 ZA67 ZB13 ZC02 ZC13 4C088 AB51 AC04 BA08 BA14 CA03 ZA14 ZA02 Z13 ZA67 ZB13 ZC13

Claims (7)

[Claims] Claims: 1. An unripe fruit of apple, pear or peach, which is squeezed and / or extracted and a fraction thereof is purified, and its component composition is a simple polyphenol compound, a caffeic acid derivative or a p-coumaric acid derivative. A polyphenol mixture comprising flavan-3-ols, flavonols, dihydrochalcones, and condensed tannins which are regular polymers of catechins as the high molecular weight polyphenol compounds. 2. An unripe fruit of apple, pear or peach, which is squeezed and / or extracted and purified, and whose component composition is a simple polyphenol compound as a caffeic acid derivative, p-coumaric acid derivative, flavan-3- An antioxidant comprising, as active ingredients, a polyphenol fraction containing alls, flavonols, dihydrochalcones, and condensed tannins, which are regular polymers of catechins as high molecular polyphenol compounds. 3. An unripe fruit of apple, pear or peach, which is squeezed and / or extracted and purified, and whose component composition is a simple polyphenol compound, a caffeic acid derivative, a p-coumaric acid derivative, a flavan-3-acid. Angiotensin I, including alls, flavonols, dihydrochalcones, and condensed tannins, which are regular polymers of catechins as high molecular polyphenol compounds
An antihypertensive comprising a polyphenol fraction having a converting enzyme inhibitory activity as an active ingredient. 4. An unripe fruit of apple, pear or peach, which is squeezed and / or extracted and purified, and whose component composition is a simple polyphenol compound as a caffeic acid derivative, a p-coumaric acid derivative, or flavan-3-one. Anti-mutation containing a polyphenol fraction having mutagenicity-suppressing activity as an active ingredient, including condensed tannins which are regular polymers of catechins as ols, flavonols, dihydrochalcone, and high molecular polyphenol compounds Genogenic agents. 5. An unripe fruit of apple, pear or peach, which is squeezed and / or extracted and purified, and whose component composition is a simple polyphenolic compound such as a caffeic acid derivative, a p-coumaric acid derivative, or flavan-3-one. Alls, flavonols, dihydrochalcones, and polyphenol fractions having a hyaluronidase inhibitory action and a histamine release inhibitory action, including condensed tannins which are regular polymers of catechins as high molecular polyphenol compounds, as active ingredients Allergy suppressant. 6. An unripe fruit of apple, pear or peach, which is squeezed and / or extracted and purified, and whose component composition is a simple polyphenol compound as a caffeic acid derivative, a p-coumaric acid derivative, or a flavan-3-acid. An anti-cariogenic agent comprising, as an active ingredient, a polyphenol fraction having glucosyltransferase inhibitory activity, which comprises a phenol, a flavonol, a dihydrochalcone, and a condensed tannin which is a regular polymer of catechins as a high molecular polyphenol compound. . 7. An unripe fruit of apple, pear or peach, which is squeezed and / or extracted and purified, and whose component composition is a simple polyphenolic compound such as a caffeic acid derivative, a p-coumaric acid derivative, or flavan-3-one. Polyphenol fractions having a deodorizing action on malodorous substances and an inhibitory action on malodorous substance production, including alls, flavonols, dihydrochalcones, and condensed tannins which are regular polymers of catechins as high molecular polyphenol compounds. A deodorant containing, as an active ingredient.