JP2001218563A - Propolis composition and method for producing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a propolis composition capable of quickly and efficiently absorbing its active ingredient which inhibit the onset and advance of diabetic complication into the human body and effectively exerting its action, and to provide a method for producing the propolis composition. SOLUTION: This propolis composition is such as to contain phenolic organic acids as an active ingredient obtained by water-extraction from a raw material propolis, and have an action to inhibit the onset and advance of diabetic complication. The concentration of the phenolic organic acids contained in this propolis composition is preferably 1.5 to 8 times higher than that of the phenolic acids contained in a propolis obtained by water-extraction. This propolis composition is obtained by the following processes: extracting an active ingredient from a raw material propolis through water-extraction; and transferring the thus obtained active ingredient which has an action to inhibit an onset and progress of diabetic complication into water.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a propolis composition which has an effect of suppressing the onset and progress of diabetic complications and is used in medicines and health foods, and a method for producing the same.

[0002]

2. Description of the Related Art In recent years, lifestyle-related diseases such as obesity, hyperlipidemia, diabetes, and hypertension have increased due to changes in eating habits. Above all, diabetes has increased about 30 times in the last 30 years, and it is said that one out of every ten people belongs to borderline diabetes, which is generally called a reserve diabetes army. Diabetes, as well as the disease itself, needs to be well aware of the dangers of its associated complications and control their onset and progression. The mechanism of the onset of diabetic complications is complex and diverse, but it is mainly caused by hyperglycemia and associated metabolic abnormalities.

[0003] Aldose reductase (aldose reductase, hereinafter referred to as AR), which is an enzyme that converts glucose or galactose into a polyol (such as sorbitol), is mainly present in peripheral nerves and the lens. In a hyperglycemic state such as diabetes, the amount of sorbitol produced increases with an increase in glucose, and the accumulation of sorbitol is one of the causes of cataract and neuropathy, which are diabetic complications.

[0004] The onset of diabetic cataract, which is one of the diabetic complications, has been studied in detail.
The osmotic pressure theory of polyols involving is predominant. In addition, other various factors are considered to be related to reduction of nicotinamide adenine dinucleotide phosphate (hereinafter referred to as NADPH) and reduced glutathione (hereinafter referred to as GSH), peroxidation of lens membrane, and the like. Have been.
In particular, GSH plays an important role in maintaining the transparency of the lens, and a dramatic decrease in GSH is said to be one of the causes of cataract.

[0005] For the purpose of preventing or delaying the onset and progress of these complications, AR inhibitors have been developed and are limited to diabetic neuropathy, but as therapeutic agents for diabetic complications. Some are commercially available. In addition, as natural products having an AR-inhibiting action, there are reported Megurinoki, Arugiku and the like, and it is considered that the main active ingredients are carboxylic acids and flavonoids.

It has been reported that some flavonoids contained in propolis have AR inhibitory activity. In addition, it was reported that a component extracted from a hydrophilic organic solvent such as ethanol contained in propolis was effective for diabetic cataract in a model experiment using rats (Medicine and Biology. 133 (2), 41- 43, 1996).
Thus, propolis has attracted attention as a natural product containing an active ingredient that suppresses the onset and progress of diabetic complications.

[0007]

However, components extracted with flavonoids and hydrophilic organic solvents contained in propolis have a drawback that they are hardly absorbed into the body because of their low solubility in water. For this reason, it has been difficult to effectively exert the effects of these active ingredients on the onset and progress of diabetic complications.

The present invention has been made by focusing on the problems existing in the prior art as described above. The purpose is to provide a propolis composition that can rapidly and efficiently absorb an active ingredient that suppresses the onset and progression of diabetic complications into the body and that can effectively exert its action. To provide.

[0009]

Means for Solving the Problems In order to achieve the above object, the propolis composition according to the first aspect of the present invention contains an active ingredient that is extracted from raw propolis with water, and produces diabetic complications. And have the effect of suppressing progress.

[0010] The propolis composition according to the second aspect of the present invention is the propolis composition according to the first aspect, wherein the active ingredient is a phenolic organic acid. The gist is to make the concentration of the phenolic organic acids contained in the water-extracted propolis obtained by extraction 1.5 to 8 times.

According to a third aspect of the present invention, there is provided a method for producing a propolis composition, comprising extracting an active ingredient having an action of suppressing the onset and progress of diabetic complications from a raw propolis by extracting the raw propolis with water. The point is to shift to.

[0012]

Embodiments of the present invention will be described below in detail. The propolis composition contains an active ingredient extracted from the raw propolis with water, and has an effect of suppressing the onset and progress of diabetic complications such as diabetic cataract and diabetic neuropathy.

[0013] The raw propolis mass as a raw material propolis is a natural resinous mass produced by honeybees.
It is formed from bee gland material and the like. In addition,
The origin of the propolis mass used here may be any of China, Brazil, European countries, Oceania countries, and the United States.

The active ingredient is contained in the water-soluble ingredient extracted from the raw propolis mass with water, and the active ingredient itself has an action of suppressing the onset and progress of diabetic complications. Specific examples of the active ingredient include phenolic organic acids such as coumaric acid, cinnamic acid, chlorogenic acid, caffeic acid, and ferulic acid.

The form of the propolis composition is not particularly limited, and may be in the form of a liquid such as an aqueous solution or a powder. Next, a method for producing the above-mentioned propolis composition will be described.

The aqueous propolis composition is obtained by adding water to the raw propolis mass and stirring to transfer the active ingredients contained in the raw propolis mass to the aqueous phase, and then removing the residue by filtration. Can be At this time,
The original propolis mass may be used after washing with alcohol such as ethanol in advance, or may be used without washing.

The powdery propolis composition is obtained by removing water by a method such as freeze-drying or spray-drying as it is or after concentrating the aqueous propolis composition obtained as described above to an appropriate concentration. Is obtained by

The propolis composition in the form of an aqueous solution is obtained by mixing the powdery propolis composition obtained as described above with:
It can also be obtained by dissolving in water again. In the present specification, each of the above-mentioned propolis compositions obtained by extracting the original propolis mass with water is referred to as “water-extracted propolis”. These water-extracted propolis may of course be used as they are for pharmaceuticals and health foods, but it is preferable to use them for pharmaceuticals and health foods after improving the concentration of the active ingredient by purification. For this purification, an ethanol addition method, a solvent extraction method, a method using a gel filtration carrier, or the like can be used. Purification is performed by one kind of method selected from these methods, or by a combination of two or more kinds of methods.

In the ethanol addition method, first, an appropriate amount of ethanol is added to a liquid propolis composition or a solution prepared by dissolving a powdery propolis composition in water or aqueous ethanol. Then, a precipitate is generated in the solution. The precipitate is removed by standing or centrifuging, and only the supernatant fraction is collected, whereby a purified propolis composition can be obtained. By this purification, the concentration of the phenolic organic acids contained in the purified propolis composition becomes 1 to 2 times the concentration of the phenolic organic acids contained in the propolis composition before purification. In particular, when the water-extracted propolis is purified by this method, the concentration of the phenolic organic acids contained in the purified propolis composition is 1.5 to 2 times that of the water-extracted propolis. The purified propolis composition is in the form of an aqueous solution in which the supernatant fraction is obtained as it is or ethanol is distilled off by distilling the supernatant fraction,
Alternatively, they are used in the form of a powder obtained by drying them.

In this ethanol addition method, the amount of ethanol added is such that the final concentration of ethanol is 30 to 80 vol / vo.
It is preferably set so as to be 1%, and more preferably 60 to 80 vol / vol% in consideration of the yield and the desired activity.

In the solvent extraction method, an organic solvent or the like is added to a liquid propolis composition or a solution prepared by dissolving a powdery propolis composition in water or aqueous ethanol, and the organic solvent or the like is utilized by utilizing the polarity of the substance. By transferring the active ingredient to the above phase, a purified propolis composition is obtained. As a result of this purification, the concentration of the phenolic organic acids contained in the purified propolis composition is 1% of the concentration before the purification.
Up to 4 times. In particular, when the water-extracted propolis is purified by this method, the concentration of the phenolic organic acids contained in the purified propolis composition is 2 to 4 times that in the case of the water-extracted propolis. Therefore, when the water-extracted propolis is purified by the ethanol addition method and further purified by this solvent extraction method, the concentration of the phenolic organic acids contained in the purified propolis composition is 3 to 8 in the case of the water-extracted propolis. Double.

In the solvent extraction method, the organic solvent may be an organic solvent such as ethyl ether, chloroform or ethyl acetate. In addition, when an acidic substance is extracted, a saturated sodium hydrogen carbonate solution (aqueous sodium bicarbonate) may be used as an aqueous phase. pH 8 ~
9, a high-concentration phosphate buffer or the like is used.

In the method using a gel filtration carrier, a column filled with a gel filtration carrier such as Sephadex is first washed with a hydrous ethanol solution acidified with hydrochloric acid, acetic acid or the like. Next, when a liquid propolis composition or a solution obtained by dissolving a powdery propolis composition in water or aqueous ethanol is added to the column, the phenolic organic acids are adsorbed to the column. Subsequently, when the aqueous ethanol solution is added to the column, the adsorbed phenolic organic acids are eluted, and the eluate is collected to obtain a purified propolis composition.

By this purification, the concentration of the phenolic organic acids contained in the purified propolis composition becomes 1 to 4 times the concentration before the purification. In particular, when the water-extracted propolis is purified by this method, the concentration of the phenolic organic acids contained in the purified propolis composition is 2 to 4 times that in the case of the water-extracted propolis. Therefore, when the water-extracted propolis is purified by the above-mentioned ethanol addition method and further purified by the method using the gel filtration carrier, the concentration of the phenolic organic acids contained in the purified propolis composition is the same as that in the case of the water-extracted propolis. It becomes 3 to 8 times. In addition,
Propolis composition after purification, as obtained eluate,
Alternatively, the eluate may be in the form of an aqueous solution from which ethanol has been distilled off, or in the form of a powder obtained by drying them.

In the method using the gel filtration carrier,
The concentration of hydrochloric acid or acetic acid in the acidic aqueous ethanol solution previously added to the column is preferably 1 to 5 vol / vol%, and the concentration of ethanol is preferably 20 to 30 vol / vol%. Further, the concentration of ethanol in the aqueous ethanol solution added to the column later is preferably from 60 to 80 vol / vol%.

In each of the above-mentioned purification methods, when the powdered propolis composition is dissolved in water to prepare a solution, the concentration of the powdered propolis composition is preferably 0.1 to 10% by weight. , More preferably 1 to 3% by weight. When the supernatant fraction obtained by the ethanol addition method is used as the liquid propolis composition, its concentration is preferably set to 0.1 to 10% by weight, more preferably 1 to 3% by weight.

As described above, according to this embodiment, the following effects are exhibited. -The active ingredient contained in the propolis composition of the embodiment, which suppresses the onset and progress of diabetic complications, is contained in the component extracted from the propolis raw mass with water. For this reason,
This active ingredient has higher solubility in water than components extracted with a hydrophilic organic solvent and flavonoids, and is quickly and efficiently absorbed into the body. Therefore, the action of suppressing the onset and progress of diabetic complications can be effectively exerted.

The water-extracted propolis is obtained by extracting a raw propolis mass with water, and therefore contains a small amount of unnecessary components contained in the raw propolis mass and contains a large amount of active ingredients. For this reason, it is possible to exert an effect of suppressing the onset and progress of diabetic complications even with a small amount of intake compared to the related art. That is, it is possible to easily and surely take an amount of the active ingredient necessary for exerting the effect of suppressing the onset and progress of diabetic complications. Further, by purifying the water-extracted propolis, the concentration of the phenolic organic acids contained in the propolis composition is increased to 1.5 to 8 times the concentration of the phenolic organic acids contained in the water-extracted propolis composition. It is possible.
Therefore, the required amount of the active ingredient can be more easily and reliably taken.

The propolis composition of the embodiment is produced by extracting raw propolis with water and transferring the active ingredient from the original propolis mass to water. For this reason,
An active ingredient can be rapidly and efficiently absorbed into the body, and a propolis composition capable of effectively exerting its action can be easily and reliably produced.

[0030]

The above embodiment will be described more specifically with reference to examples and test examples. (Example 1) 10 kg of a raw propolis mass produced in Brazil was pulverized with a pulverizer, and 95 vol / vol% of ethanol 20 was added thereto.
After adding 0 liter, the mixture was left at room temperature for 4 days with occasional stirring. Thereafter, the supernatant was separated and removed by filtration to obtain an ethanol-washed propolis mass. next,
15 liters of warm water is added to 2.5 kg of the ethanol-washed propolis mass, and the mixture
After the time extraction, solid-liquid separation was performed with a bucket type centrifuge to obtain a separated liquid. Subsequently, diatomaceous earth 1.7 was added to the separated liquid.
After adding 5 kg and stirring, filtration was performed to obtain a filtrate.
The filtrate was concentrated and dried by a freeze vacuum dryer to obtain 110 g of a dry powder of water-extracted propolis.

Example 2 To 18 g of the dry powder of water-extracted propolis obtained in Example 1, 900 ml of warm water at 60 ° C. was added and stirred for 1 hour, and then 2100 ml of ethanol was added and the mixture was further stirred for 2 hours. Then, the water-ethanol solution was allowed to stand at room temperature overnight to precipitate the precipitate, and only the supernatant fraction was collected. The supernatant fraction was concentrated and dried under reduced pressure to obtain 9.5 g of dry powder.

Example 3 1750 ml of the supernatant fraction before concentration obtained in Example 2 was concentrated to 350 ml and adjusted to pH 3 with hydrochloric acid to obtain a preparation. 360 ml of ethyl acetate was added to this prepared solution, and the mixture was separated into an aqueous phase and an ethyl acetate phase with a separating funnel. The obtained aqueous phase was again added with 360 ml of ethyl acetate and separated twice in the same manner. Then, the ethyl acetate phases were combined, concentrated to 120 ml, and adjusted to pH 9 with sodium hydroxide to obtain a prepared solution. Subsequently, 125 ml of a saturated sodium bicarbonate solution was added to the prepared solution, and the mixture was separated into a sodium bicarbonate solution phase and an ethyl acetate phase with a separatory funnel. The operation of separation was performed twice more. Then, the saturated sodium bicarbonate solution phases were combined, and this was adjusted to pH 2.8 with hydrochloric acid to obtain a prepared solution. Further, 380 ml of ethyl acetate was added to the prepared solution, and the mixture was separated into a sodium bicarbonate solution phase and an ethyl acetate phase by using a separating funnel.
The same operation was performed by adding ml. Then, the ethyl acetate phases were combined, concentrated, and dried under reduced pressure to obtain 1.7 g of dry powder.

Example 4 A gel filtration column using Sephadex LH-20 as a carrier was prepared using 100 ml of glass burette, and the composition was ethanol / distilled water / acetic acid = 25/75/1 (vol / vol). %) In the column. Then, 300 mg of the dry powder obtained in Example 1 was dissolved in 3 ml of an elution solvent, added from the top of the column, and eluted with 100 ml of the elution solvent to adsorb phenolic organic acids and the like to the column. The eluate at this time is referred to as fraction-1 (hereinafter, referred to as Fr-1).
Here, elution was carried out by changing the elution solvent to a 70 vol / vol% ethanol solution, and the elution was performed at 60 ml, 40 ml, 40 ml, 8 ml, and 80 ml.
0 ml of Fr-2, 3, 4, and 5 were collected. Fr-1 ~
5 was concentrated and dried under reduced pressure, and 211 mg, 14 mg, 78 mg, 24 mg
mg and 11 mg were obtained.

(Test Example 1) The dried powders obtained in Examples 1 to 4 were analyzed by high performance liquid chromatography (hereinafter referred to as HPLC) in order to examine the content of phenolic organic acids. The area and the total area of the peaks were measured. Table 1 shows the results.

The HPLC analysis conditions are as follows. Column: ODS column (SHISEIDO C18 AG120Å 4.6 × 15
0 mm), mobile phase: methanol / distilled water / acetic acid = 25/7
5/1 (vol / vol%), temperature: room temperature, flow rate: 0.8 ml
/ Min, detector: ultraviolet ray (UV) 315 nm, injection amount: 5 μl, sample: 1% solution of each dry powder HPLC analysis using chlorogenic acid, caffeic acid, p-coumaric acid, and ferulic acid as standard products After doing
The holding time of each standard product was 3.7 min, 5.1 respectively.
min, 8.0 min, and 9.5 min.

[0036]

[Table 1] From the results shown in Table 1, the HP of Fr-1 was found to be about Example 4.
Almost no peak corresponding to phenolic organic acids was detected by LC analysis. From this, it was found that phenolic organic acids can be adsorbed on a Sephadex column by using an elution solvent of ethanol / distilled water / acetic acid = 25/75/1 (vol / vol%). Further, Fr-2-after replacing the elution solvent with 70 vol / vol% ethanol
As shown in the analysis result of No. 5, it was found that most of the phenolic organic acids adsorbed on the column could be recovered by using the aqueous ethanol solution as the second elution solvent.

Comparing the peak areas, Example 2 is 1.4 to 2.1 times that of Example 1, and Example 3 is 1.1 to 2.1 times that of Example 1.
It was 4 to 22 times. In addition, each fraction of Example 4 was 1/1 of Example 1 for any fraction.
There was an opening due to the peak at 0 to 100 times.

Comparing the total area of the peaks, Example 2 is 1.6 times that of Example 1, Example 3 is 4.9 times that of Example 1,
In Example 4, Fr-1 was 0.03 times that of Example 1, and Fr-2
1.3 times for Fr, 2.6 times for Fr-3, Fr-
4 was 5.7 times, and Fr-5 was 3.4 times. The total area after averaging the peak areas of Fr-2 to 5 was 3.4 times that of Example 1, and the total area after averaging the peak areas of Fr-3 to 5 was also 4.0 times. Met.

Accordingly, it was shown that any of the purification methods of Examples 2 to 4 can provide a propolis composition containing a higher amount of phenolic organic acids than the propolis composition of Example 1. .

(Test Example 2) In order to examine the AR inhibitory activity of each of the dry powders obtained in Examples 1 to 3, an AR enzyme solution was prepared by the following method, and the AR inhibitory activity was measured. In addition, these preparation methods and activity measurement methods are well-known methods (Archives of Biochemistry and Biophysics, 216
(1), 337-344, 1982, The Journal of Antibiotics, 48 (1
1), 1345-1346, 1995).

Ten fresh bovine lenses were finely cut with scissors, and cooled with 5 mM PBS (Phosphate-buffered saline) containing 1 mM 2-mercaptoethanol.
A double volume was added and homogenized while cooling. This homogenate solution was centrifuged at 4 ° C. and 1.8 × 10 ⁵ m / s ² for 30 minutes, and the supernatant was collected to obtain a crude AR extract. next,
0.4 ml of AR crude extract and 1
2.0 ml of PBS containing 00 mM lithium sulfate;
0.2 ml of 1.5 mM NADPH and 0.2 ml of a solution prepared by dissolving each of the dry powders obtained in Examples 1 to 3 in water are mixed by inversion. Subsequently, after preincubating the cells at 25 ° C. for 10 minutes, 75 mM DL-
Add 0.2 ml of glyceraldehyde solution and mix by inversion. Then, for this sample solution (3.0 ml in total), the change in absorbance at 340 nm was recorded at 25 ° C. for 5 minutes. From this measurement record, the change in absorbance per minute was calculated, and the AR inhibition rate was calculated according to the following equation. Table 2 shows the results. AR inhibition rate (%) = (1− (ΔAs−ΔAb) / (ΔA
c-ΔAb)) × 100 ΔAb: change in absorbance per minute when PBS containing 100 mM lithium sulfate is used instead of the solution and DL-glyceraldehyde solution ΔAc: 100 mM lithium sulfate instead of the solution ΔAs: change in absorbance per minute of sample solution ΔBS: change in absorbance per minute of sample solution in accordance with the least squares method from the relationship between the concentration of dry powder in the sample solution and the AR inhibition rate The concentration (IC ₅₀ ) at which 50% was inhibited was determined. Table 2 also shows the results.
Shown in

[0042]

[Table 2] From the results in Table 2, the AR inhibitory activity is about 1.4 times stronger in Example 2 than in Example 1, and about 2.6 in Example 3 as in Example 1.
It was found to have twice the activity. Therefore, the propolis compositions obtained in Examples 2 and 3 have stronger AR inhibitory activity than the propolis composition of Example 1, and the intensity of this activity is proportional to the content of phenolic organic acids. Was suggested.

(Test Example 3) In order to examine the effect of the propolis composition on diabetic complications, a model experiment was conducted using rats on diabetic cataract in which AR is involved in the onset and progression.

[Preparation of Galactose-Induced Diabetic Cataract Rats] Sprague-Dawley male rats (weight around 90 g) were allowed to freely ingest the diets shown in Table 3 for 18 days in groups A to E (6 rats per group). Was. In addition, the powder feed in Table 3 is a trade name CRF- manufactured by Oriental Yeast Co., Ltd.
It is one. In addition, the amount of D-galactose and the duration of its intake in Groups A to D are sufficient conditions to cause galactose-induced diabetic cataract in rats.

[0045]

[Table 3] [Evaluation of turbidity of lens] After sufficiently dilated the eyes with 0.5% tropicamide ophthalmic solution (manufactured by Wakamoto Pharmaceutical Co., Ltd.), the lens was observed with the naked eye, and the turbidity was measured by the Sippel method (Inves method).
t.Ophthalmol., 5,568-575,1966)
ore 0, 1, 2, 3, 4). Table 4 shows the results. From Table 4 onwards, all values in the table are Mean ± SD (n = 6)
Is shown. In addition, Student t-test was used for the statistical processing. In Table 4 and subsequent figures, * in the table indicates that the significant difference from the control group A was 5% or less, and *
* Indicates that the significant difference with respect to the group A is 1% or less.

[0046]

[Table 4] From the results in Table 4, it was confirmed that the lens opacity was lower in groups B to D than in group A, and that the propolis composition had an inhibitory effect on lens opacity in galactose-induced diabetic cataract. In particular, this inhibitory effect was the strongest in group D and significantly lower than in group A. in this way,
Since the AR inhibitory activity and the opacity of the lens are inversely proportional, it was shown that the stronger the AR inhibitory activity, the stronger the effect of suppressing the opacity of the lens, that is, the onset and progress of cataract.

[Measurement of Weight of Lens] The eyeball was excised, the lens was separated therefrom, and the weight was measured. Table 5 shows the results.

[0048]

[Table 5] From the results in Table 5, the lens weight was significantly lower in the C and D groups than in the A group, but was not significantly different from that in the E group that did not induce diabetes. Judged that it was not an adverse effect.

[Measurement of GSH Content in Lens] To the separated lens, 0.5 ml of a 0.1 M sodium phosphate solution (pH 8.0) containing 0.005 M ethylenediaminetetraacetic acid was added per lens, and homogenated. Then 4
The supernatant was separated by centrifugation at 3000 rpm for 10 minutes. The GSH content in this supernatant was determined by a fluorescence method using O-phthalaldehyde (Analytical biochemistry, 74, 214-226, 1).
Table 6 shows the results measured according to 976).

[0050]

[Table 6] Generally, in the case of diabetes, it is said that there is a tendency for GSH in the lens to decrease. As shown in Table 6, results supporting this were obtained. Also, from the results in Table 6,
The GSH content per lens was significantly higher in the BD group than in the A group, indicating that the propolis composition suppressed the decrease in GSH in the lens due to diabetes. In addition, the GSH content per lens weight is significantly higher in the C and D groups than in the A group, and the C group has a higher DSH than the B group.
The group was higher. Therefore, the propolis composition of Example 3 has the strongest effect of suppressing the reduction of GSH in the lens, and it is inferred that the propolis composition of Example 2 and the propolis composition of Example 1 are in the following order.

[Measurement of GSH Content in Serum] After anesthesia with ether, blood was collected from the abdominal vena cava, and the blood was centrifuged to obtain serum. The GSH content in the serum was determined by a fluorescence method using O-phthalaldehyde (Analytical biochemistry,
74, 214-226, 1976) are shown in Table 7. The supernatant was separated from the serum deproteinized with three times the amount of ethanol, and the supernatant treated with O-phthalaldehyde was used for the measurement.

[0052]

[Table 7] Generally, in the case of diabetes, it is said that there is a tendency that the GSH in blood tends to increase, but as shown in Table 7, results supporting this were obtained. Also, from the results in Table 7, B
GSH in serum was lower in Groups D to D than in Group A, and was significantly lower in Group D than in Group A. Therefore, G in blood
It can be said that the propolis composition of Example 3 has the strongest effect of suppressing the increase in SH.

As described above, since the propolis composition suppresses the decrease of GSH in the lens and the increase of GSH in the blood, which is said to be the cause of cataract, it has the effect of suppressing the onset and progress of diabetic cataract. It was shown that. Further, the effect was strongest in the propolis composition of Example 3 and then in the order of Example 2 and Example 1. Therefore, it is considered that the inhibitory effect is proportional to the AR inhibitory activity and the phenolic organic acid content. Can be

[Measurement of Lipid Peroxide Content] The lipid peroxide content in the lens and serum was measured using a lipid peroxide measurement kit (Liquid peroxide-Test Wako, manufactured by Wako Pure Chemical Industries, Ltd.). Table 8 shows the results of measuring the lipid peroxide content in the lens, and Table 9 shows the results of measuring the lipid peroxide content in the serum.
Shown in

[0055]

[Table 8]

[0056]

[Table 9] From the results in Table 8, the lipid peroxide content in the lens was B to D
It was lower in the group than in the group A, and was suppressed to the same level as in the group E which did not induce diabetes.

From the results shown in Table 9, no difference was observed in the serum lipid peroxide between the groups A and E, and no effect was observed due to galactose-induced diabetes. Further, no influence by the propolis composition was observed.

As described above, in the galactose-induced diabetic cataract, the lipid peroxide content in the serum was not changed, but the lipid peroxide in the lens was increased, and the propolis composition caused this increase. It was shown that the effect was suppressed to the normal level.

Next, the technical ideas that can be grasped from the above embodiment will be described below. (A) Ethanol is added to a water-extracted propolis solution obtained by water-extracting the raw material propolis to form a precipitate,
The method for producing a propolis composition according to claim 3, wherein the supernatant fraction is collected. With such a configuration, the amount of the active ingredient in the propolis composition can be increased.

(A) The propolis according to (A), wherein a solvent is added to the supernatant fraction, the active ingredient is transferred from the supernatant fraction to the solvent, and the solvent is collected. A method for producing the composition. With such a configuration, the amount of the active ingredient in the propolis composition can be further increased.

(C) The above-mentioned supernatant fraction is added to a gel filtration carrier, the active ingredient is adsorbed on the gel filtration carrier, then eluted with an extraction solvent, and the eluate is collected. A method for producing a propolis composition according to the above (A).
With such a configuration, the amount of the active ingredient in the propolis composition can be further increased.

[0062]

The present invention is configured as described above, and has the following effects. ADVANTAGE OF THE INVENTION According to the propolis composition of the invention of Claim 1, the active ingredient which suppresses the onset and progress of diabetic complications can be quickly and efficiently absorbed into the body, and its action is effectively exhibited. Can be done.

According to the propolis composition of the second aspect of the present invention, in addition to the effect of the first aspect of the present invention, an effective amount of the propolis composition capable of exerting an inhibitory effect on the onset and progression of diabetic complications. Ingredients can be easily and reliably taken.

According to the method for producing a propolis composition according to the third aspect of the invention, the propolis composition according to the first or second aspect of the invention can be easily and reliably produced.

Continued on the front page (72) Inventor Satoshi Mishima 1-1-1, Kano Sakuradacho, Gifu-shi Api Co., Ltd. F-term (reference) 4B018 MD78 ME03 MF01 4B041 LD06 LK35 LP05

Claims (3)

[Claims] 1. A propolis composition comprising an active ingredient extracted from raw propolis with water and having an action of suppressing the onset and progress of diabetic complications. 2. The active ingredient is a phenolic organic acid, and the content of the phenolic organic acid is determined by adjusting the content of the phenolic organic acid to 1.times. The concentration of the phenolic organic acid contained in the water-extracted propolis obtained by extracting the raw propolis with water. The propolis composition according to claim 1, wherein the ratio is 5 to 8 times. 3. A method for producing a propolis composition, comprising extracting an active ingredient having a function of suppressing the onset and progress of diabetic complications from a raw propolis by extracting the raw propolis with water. .