WO2020255502A1 - Labdane-type diterpene compound-containing stabilizing formulation for oral administration, and production method therefor - Google Patents

Abstract

The present invention pertains to: a labdane-type diterpene-containing stabilizing formulation for oral administration; and a production method therefor. This stabilizing formulation for oral administration is characterized in that a labdane-type diterpene compound is contained in plant oil. Moreover, this method for producing a stabilizing formulation is characterized by including: (A) preparing an extraction mixture by using a hydrophobic organic solvent to extract a plant containing a labdane-type diterpene compound, and then adding plant oil to the obtained extract, or (B) preparing an extraction mixture by using a mixture of a hydrophobic organic solvent and plant oil to extract a plant containing a labdane-type diterpene compound; and removing the hydrophobic organic solvent from the obtained extraction mixture to obtain a stabilizing formulation which is for oral administration and in which the labdane-type diterpene compound is contained in the plant oil.

Description

Stabilized preparation for oral administration containing a labdane-type diterpene compound and a method for producing the same.

The present invention relates to a stabilized preparation for oral administration containing a labdane-type diterpene compound used in various uses such as pharmaceuticals, functional foods, and health foods, and a method for producing the same.

Labdane-type diterpenes compounds are known to exhibit various pharmacological actions, and are used as active ingredients in skin-beautifying compositions and synovial growth inhibitors. These labdane-type diterpenes compounds are contained in plants, but there is a problem that the content in plants is low, so they are taken orally as they are and ingested in an amount sufficient to obtain the desired effect. It is difficult to do so, so it is extracted and used.

As the extraction method, a method using hexane (Patent Document 1), a method by hexane or supercritical extraction method (Patent Document 2), polar and non-polar such as ethyl acetate ester, pentane, hexane, acetone, methyl alcohol and ethyl alcohol. A method using an organic solvent (Patent Document 3) is known.

Japanese Unexamined Patent Publication No. 2006-24168 Japanese Unexamined Patent Publication No. 2013-60403 Japanese Unexamined Patent Publication No. 2014-152165

As described above, the labdane-type diterpene compound is extracted from a plant containing the labdane-type diterpene compound and used, but it is inevitable to concentrate the extract after the extraction.

However, the labdane-type diterpene compounds are sensitive to heat, and when the extract is concentrated, the content of the labdane-type diterpenes compounds decreases, causing coloring and the like.

For example, when a labdane-type diterpene compound is extracted with hexane and then the extract is concentrated by heating, the extract is colored blackish brown and becomes resinous even when concentrated under reduced pressure. Insoluble in solvents such as water, alcohol and hexane.
The colored or resinous extract has a problem that the active ingredient is reduced.

It is considered that this is because the labdane-type diterpenes compound contains highly reactive groups that serve as active centers, such as aldehyde groups and methylene groups, and thus easily binds to adjacent molecules to cause condensation or polymerization. ..

On the other hand, for example, hexane must not be present in foods according to the Food Sanitation Law. Hexane is oxidized in the body as acute toxicity such as headache and mild anesthetic action, and as chronic toxicity, 2,5-hexanedione. Since multiple neuropathy such as difficulty walking due to the formation of hexane is known, removal after extraction cannot be avoided in hexane extraction.

Further, when extracted with water or ethyl alcohol, the labdane-type diterpene compound is fat-soluble, so that the content in the extract is not sufficient, and a desired content of the labdane-type diterpene compound can be obtained. Excessive heat concentration is required for this, and colored substances, polymers, and condensates increase.

There is no particular problem if a labdane-type diterpene compound is obtained at the laboratory level, but in actual production, the decrease in active ingredient due to this coloring or polycondensation cannot be overlooked.

Furthermore, if the supercritical extraction method is adopted, the above-mentioned coloring can be avoided, but it is difficult to set the extraction conditions, a special device is required, the operation is complicated, and there is a problem in terms of cost.

As a result of diligent research, the present inventors have added vegetable oil to an organic solvent extract containing a labdane-type diterpene compound, and then concentrated the extract to compress the labdane-type diterpene compound in the extract, and based on this. As a result of suppressing coloring and removing the organic solvent as an extraction solvent, it has been found that a vegetable oil containing a high concentration of labdane-type diterpene compounds that can be orally ingested can be obtained, and the present invention has been completed.

An object of the present invention is to provide a stabilized preparation that stably contains a labdane-type diterpene compound and can be orally ingested, and a method for producing the same.

The present invention is a stabilized preparation for oral administration, which comprises a labdane-type diterpene compound contained in a vegetable oil.

Further, the present invention is characterized in that the labdane-type diterpene compound is extracted from a plant containing the labdane-type diterpene compound by an organic solvent, and the vegetable oil does not substantially contain the organic solvent.

Further, in the present invention, the labdane-type diterpenes compounds are (E) -8β (17) -labd-12-ene-15,16,17-trial, (E) -8β (17) -epoxylabd-12-. One selected from en-15,16-diene, 16-oxo-8 (17), 12 (E) -labdane-icic acid and labdane-8 (17), 12-diene-15,16-dial It is characterized by the above.

Further, the present invention is characterized in that the vegetable oil is rice bran oil.

Further, in the present invention, a plant containing (A) labdane-type diterpene compounds is extracted with an organic solvent, and then vegetable oil is added to the obtained extract to prepare an extraction mixture, or (B) labdane-type diterpenes. A plant containing the compound is extracted with a mixture of an organic solvent and a vegetable oil to obtain an extraction mixture, and the organic solvent is removed from the obtained extraction mixture to contain a labdane-type diterpene compound in the vegetable oil for oral administration. It is a method for producing a stabilizing preparation for oral administration, which comprises obtaining a stabilizing preparation.

Further, the present invention is characterized in that the plant containing the labdane-type diterpenes compound is turmeric, and the organic solvent is at least one selected from ethyl alcohol, methyl alcohol, acetone, hexane, cyclohexane, heptane and diethyl ether. And.

Further, in the present invention, a plant containing (C) a labdane-type diterpen compound is extracted with an organic solvent, and then vegetable oil is added to the obtained extract to prepare an extraction mixture, or (D) labdane-type diterpenes. A method for concentrating a Labdane-type diterpene compound, which comprises extracting a plant containing a compound with a mixture of an organic solvent and a vegetable oil to obtain an extraction mixture, concentrating the obtained extraction mixture, and removing the organic solvent. is there.

Further, the present invention is a composition for oral ingestion containing a labdane-type diterpene compound, which comprises a labdane-type diterpene compound contained in a vegetable oil.

According to the present invention, the stabilized preparation contains a labdane-type diterpene compound at a high concentration and can suppress coloring and polycondensation of the labdane-type diterpene compound which is an active ingredient, and thus contains an active ingredient. Does not decrease.

For example, the hyaluronidase inhibitory activity, which is one of the important actions of the labdane-type diterpenes compound of the present invention, does not change.

Further, since the medium of the stabilized preparation for oral administration of the present invention is vegetable oil, it can be taken as it is or further formulated and taken orally.

Further, according to the method for producing a stabilized preparation for oral administration of the present invention, it is only necessary to add vegetable oil to an organic solvent which is an extraction solvent at the time of extraction of a labdane-type diterpene compound, or to an organic solvent extract after extraction. Therefore, no special device or expensive solvent is required, the existing device can be used as it is, and the coloring and decompression of the labdane-type diterpene compound can be easily suppressed.

Further, according to the method for producing a stabilized preparation for oral administration of the present invention, all the labdane-type diterpenes compounds in the extract extracted with the organic solvent are contained in the vegetable oil, so that the amount of the vegetable oil The content of the labdane-type diterpenes compound in the vegetable oil can be adjusted by adjusting the above.

Further, according to the concentration method of the present invention, since vegetable oil is only added to the extract before concentration, no special equipment or expensive solvent is required, and the existing equipment can be used as it is, and a labdane type can be easily used. It is possible to prevent a decrease in activity and coloring of diterpenes compounds.

The stabilized preparation for oral administration of the present invention is a preparation in which a labdane-type diterpene compound is contained in a vegetable oil. More specifically, it is a preparation in which only the organic solvent is removed from a mixture of an organic solvent containing a labdane-type diterpene compound and a vegetable oil, and the labdane-type diterpene compound is contained in the vegetable oil as a residue.

The stabilized preparation for oral administration of the present invention is an oily preparation containing a labdane-type diterpene compound and exhibiting a yellow to yellowish brown color, and contains 0.5 to 3.0%, preferably 1. Contains 0-2.0%.

In the present invention, the labdane-type diterpene compound includes various compounds, and as a typical labdane-type diterpene compound, (E) -8β (17) -labd-12-ene-15,16,17-trial , (E) -8β (17) -epoxylabd-12-ene-15,16-diar, 16-oxo-8 (17), 12 (E) -labdaziene-15-euic acid and labdane-8 (17). , 12-diene-15,16-jiar and the like.

The content of the compound in the vegetable oil varies depending on the extraction raw material, the extraction solvent, the extraction conditions, the amount of the vegetable oil added before concentration, and the like. For example, (E) -8β (17) -labd-12-ene-15. , 16,17-Trial 0.025-0.18%, (E) -8β (17) -epoxylabd-12-ene-15,16-diene 0.025-0.12%, 16-oxo -8 (17), 12 (E) -labdadiene-15-oic acid was 0.05-0.3% and rabda-8 (17), 12-diene-15,16-diar was 0.4-2. It is 4%.

In the present invention, these labdane-type diterpenes compounds may be free compounds or various salts. Further, all four types may be contained, or any one or more types may be contained. In the case of a mixture of four kinds of compounds, four kinds of equal amounts mixture may be used, two to three kinds may be equal amount mixtures, and the remaining two to one kinds may have different contents, and further. May have different content ratios of all four types.

Further, in the stabilized preparation for oral administration of the present invention, various essential oil components derived from the extraction raw material, for example, pharmacology such as curcumin, turmeron, cineole, curcumenone, curcumol, elemen, paramethitolylcapinol, flavonoids, azulene and the like. It may contain an active essential oil component.

In the present invention, the vegetable oil may be any vegetable oil that dissolves the labdane-type diterpene compound and the essential oil component derived from the extraction raw material, can be taken orally, and does not azeotrope with an organic solvent.

Examples of such vegetable oils include coconut oil, corn oil, cottonseed oil, olive oil, palm oil, peanut oil, rapeseed oil, rice oil, sesame oil, soybean oil, sunflower oil, almond oil, cashew nut oil, hazelnut oil, macadamia oil and the like. ..

All of these vegetable oils contain unsaturated fatty acids having 15 to 16 carbon atoms, and all of them have the solubility of the labdane-type diterpene compounds. Further, these vegetable oils can be easily mixed with the organic solvent of the extraction solvent described later, and at the time of concentration, only the solvent can be evaporated and removed from the mixed solution.

Among these, preferable vegetable oils include rice oil, soybean oil, and corn oil, and commercially available products can be used. Examples of the commercially available products include rice bran oil (trade name, manufactured by Oriza Yuka Co., Ltd.). Examples include soybean oil (trade name, manufactured by Ajinomoto Co., Ltd.) and corn salad oil (trade name, manufactured by Tsuji Oil Co., Ltd.).

Examples of more preferable vegetable oils include rice bran oil, and refined rice oil from which free fatty acids and waxes have been removed is the most preferable.

These vegetable oils may be used alone or in combination of one or more.

The stabilized preparation for oral administration of the present invention thus obtained can be orally ingested in a state in which a labdane-type diterpene compound is contained in a vegetable oil.

Further, by encapsulating and filling a soft capsule or a capsule with a vegetable oil containing a labdane-type diterpene compound, the preparation can be easily taken.

In the stabilized preparation for oral administration of the present invention, a labdane-type diterpene compound is extracted from a plant containing the labdane-type diterpene compound with an organic solvent, and then the organic solvent is removed. Does not contain organic solvents.

Here, the fact that the organic solvent is not contained means that the organic solvent is not detected (below the detection limit) by a known detection method, for example, GC-MS (gas chromatograph mass spectrometer).

In the present invention, the labdane-type diterpene compound extracted by the organic solvent is transferred to the above-mentioned vegetable oil as the organic solvent is removed by concentration. Therefore, the organic solvent is selected according to the extract and the concentration conditions. Further, by selecting the vegetable oil as a medium of the oral administration preparation and the amount of the vegetable oil added thereto, a stable oral administration preparation having a desired concentration of the labdane-type diterpene compound can be obtained regardless of the vegetable oil. ..

In the stabilized preparation for oral administration of the present invention, (A) a plant containing a labdan-type diterpene compound is extracted with an organic solvent, and then vegetable oil is added to the obtained extract to prepare an extraction mixture. A method for removing an organic solvent from a method (hereinafter referred to as method A) or (B) a plant containing a labdan-type diterpene compound is extracted with a mixture of an organic solvent and a vegetable oil, and the organic solvent is extracted from the obtained extraction mixture. It can be produced by a method of removing (hereinafter, referred to as method B).

In the present invention, examples of plants containing a labdan-type diterpene compound as an extraction raw material include plants of the family Zingiberaceae, and examples of plants of the family Zingiberaceae include plants belonging to the genus Ginger, plants belonging to the genus Ginger, and plants belonging to the genus Zingiberaceae. , Plants belonging to the genus Zingiberaceae.

The plant belonging to the plants belonging to the genus Hidden-lilies, Hidden-lilies, Hidden-lilies and Hidden-lilies may be any as long as it contains a Labdan-type diterpene compound, but to give a specific example, the genus Hidden-lilies Plants belonging to the group include white turmeric, purple turmeric, spring turmeric, autumn turmeric (Curcuma caeasia, Curcuma zedoraia, Curcuma aromatica, Curcuma longa), black turmeric (Kaempferia paraviflora), mango turmeric, etc.

In addition, as plants belonging to the genus Zingiber, there are myoga (Zingiber mioga), Oyama ginger (Zingiber spectabile), ginger (Zingiber officinale), red ginger (Zingiber officinale Rubra) and the like.

In addition, examples of plants belonging to the genus Alpinia include Alpinia japonica and shell ginger (Alpinia speciosa), and plants belonging to the genus Hedychium include Sanna (Hedychium spicatum) and Hanashukusha (Hedychium). Can be given.

Among the above plants, white turmeric, which is known to have the highest content of labdane-type diterpenes compounds, is preferable.

The solvent is not particularly limited as long as it easily evaporates when a labdane-type diterpene compound is dissolved, mixed with the vegetable oil and concentrated.

As such a solvent, a hydrophilic organic solvent or a hydrophobic organic solvent can be used, and as the hydrophilic organic solvent, lower alcohols such as methyl alcohol, ethyl alcohol and propyl alcohol, ketones such as acetone and methyl ethyl ketone, and ethyl acetate Examples thereof include esters such as esters, formic acid ethyl esters and butyric acid ethyl esters.

Examples of the hydrophobic organic solvent include hydrocarbons such as pentane, hexane, heptane, nonane and decane, and cyclic or aromatic hydrocarbons such as cyclohexane, toluene and benzene. Of these, pentane and hexane are preferable.

A known extraction method can be adopted for extraction from plants containing Labdane-type diterpenes compounds. In both the A method and the B method, the amount of the extraction solvent may be 30 to 60 parts by mass with respect to 100 parts by mass of the plant containing the labdane-type diterpene compound.

Further, the extraction temperature can be carried out at any temperature between room temperature and the recirculation temperature of the organic solvent used in both the A method and the B method, and is, for example, in the range of 20 to 60 ° C. The extraction time is preferably 24 to 72 hours, for example. When a plant containing a labdane-type diterpene compound is immersed in an organic solvent for extraction, the organic solvent is 100 to 300 parts by mass with respect to 100 parts by mass of the plant containing the labdane-type diterpene compound. It may be used.

Specifically, an appropriate part of the plant containing the labdane-type diterpene compound, for example, rhizome, stem, leaf, flower, etc., can be used as an extraction solvent as it is or by crushing it, or by cutting it after drying or drying. It can be easily carried out by contacting and mixing with an organic solvent or a mixture of an organic solvent and a vegetable oil, transferring a labdane-type diterpene compound and an essential oil component to the extraction solvent, and then separating the extract.

Concentration should be carried out under reduced pressure to avoid coloring and polycondensation of Labdane-type diterpenes compounds.

As for the vegetable oil added to the organic solvent, all the labdane-type diterpenes compounds in the organic solvent are transferred to the vegetable oil. Therefore, the concentration of the labdane-type diterpenes compounds in the vegetable oil should be adjusted by adjusting the amount of the vegetable oil added. Can be done.

For example, when vegetable oil equivalent to half the amount of the organic solvent is added, after removing the organic solvent, the concentration of the labdane-type diterpene compounds in the vegetable oil is twice that at the time of extraction, which is equivalent to one-third. When vegetable oil is added, the concentration of labdane-type diterpene compounds in the vegetable oil becomes three times that at the time of extraction after the organic solvent is removed, so the amount of vegetable oil added should be determined in consideration of this. it can.

The concentration of the Labdane-type diterpene compound in the extract can be determined by using an analytical instrument such as gas chromatography, mass spectrometer, or liquid chromatography for the extract, so the final desired concentration should be taken into consideration. The amount of vegetable oil to be added can be determined. If the amount of the extraction solvent is large, it takes time to concentrate, and coloring and polycondensation occur. Therefore, usually, 10 to 100 parts of vegetable oil may be used with respect to 100 parts of the extraction solvent by volume.

As described above, all labdane-type diterpenes compounds in the extract are transferred to the vegetable oil.
Since the concentration is increased by adjusting the amount of vegetable oil to be smaller than the amount of organic solvent, for example, the method for producing a stabilized preparation for oral administration of the present invention should be regarded as a method for concentrating labdane-type diterpenes compounds. Can be done.

Further, at that time, the amount of colored products and polycondensation products produced is much suppressed as compared with the case of concentrating by evaporating this using only an organic solvent, so that the present invention is stable for oral administration. The method for producing the chemical preparation can be regarded as a method for preventing coloration, a method for preventing quality deterioration, or a method for preventing polycondensation of a labdane-type diterpene compound.

When the present invention is a method for concentrating a labdane-type diterpene compound, the concentration method of the present invention is to extract (C) a plant containing a labdane-type diterpene compound with an organic solvent, and then add vegetable oil to the obtained extract. Is added to the extract mixture, the extract mixture is concentrated, and only the organic solvent is removed (hereinafter referred to as method C), or (D) a plant containing a labdane-type diterpene compound is mixed with an organic solvent and vegetable oil. It can be carried out by a method (hereinafter referred to as method D) in which only the organic solvent is removed by concentrating the obtained extraction mixture and preparing an extraction mixture.

At this time, the vegetable oil to be used can be reliably concentrated by setting the amount to be smaller than the amount of the organic solvent, and for example, the desired concentration ratio can be obtained by adjusting the volume ratio of the organic solvent and the vegetable oil. ..

By carrying out both the C method and the D method according to the method for producing a stabilized preparation for oral administration, the labdane-type diterpenes compound can be concentrated efficiently and safely.

The stabilized preparation for oral administration of the present invention contains 0.1 to 0.3% of α-tocopherol, 0.5 to 5.0% of pinene, which is a monoterpene compound, and dehydro, which is a sesquiterpene compound. It is characterized by containing 3.0 to 8.0% of crudione, 1.0 to 6.5% of crudion, 2.5 to 7.0% of crucumenone, and 5.0 to 10% of cruzelenone.

Hereinafter, the present invention will be described in more detail with reference to Examples.

Each evaluation method and allowable range in the examples are as follows.
<Hyaluronidase inhibitory activity measurement method>
40 μl of a sample solution (treatment concentration: 5 mg / ml) in which an oily substance was dissolved in DMSO and 160 μl of 0.1 M acetate buffer (pH 3.8 to 4.0) were placed in a tube and mixed well, and hyaluronidase was added thereto. 100 μl of the solution (4000 units / ml) was added and incubated at 37 ° C. for 20 minutes. Next, 200 μl of Compound 48/80 solution (0.5 mg / ml) was added as a hyaluronidase activator, and the mixture was further incubated at 37 ° C. for 20 minutes.

Subsequently, 500 μl of sodium hyaluronate solution (0.8 mg / ml) was added and incubated at 37 ° C. for 40 minutes. Then, 200 μl of a 0.4N sodium hydroxide aqueous solution was added as a reaction terminator, and the mixture was cooled on ice to terminate the reaction.

After stopping the reaction, 200 μl of 0.1 M boric acid buffer (pH 9.1) was added, mixed well, heated at 100 ° C. for 5 minutes, and then cooled on ice. Subsequently, it was centrifuged at 20000 G for 10 minutes. Next, 140 μl of the supernatant after centrifugation and 600 μl of the coloring reagent were mixed well and incubated at 37 ° C. for 20 minutes. This is referred to as sample S1.

As the color-developing reagent, 25 g of p-dimethylaminobenzaldehyde was dissolved in a mixed solution of 12.5 ml of 10N hydrochloric acid and 37.5 ml of acetic acid, and a storage solution stored at 10 ° C. or lower with an expiration date of 1 month was used immediately before use. Was diluted 10-fold with acetic acid.

Similarly, DMSO was used instead of the sample solution (hereinafter referred to as "control C1"), and 0.1 M acetate buffer (pH 3.8 to 4.0) was used instead of hyaluronidase (hereinafter referred to as "control C1"). , "Sample blank SB1"), and those using DMSO instead of the sample solution and 0.1 M acetate buffer (pH 3.8-4.0) instead of hyaluronidase (hereinafter, "control blank CB1"). ") Was prepared. Hyaluronidase, Compound 48/80, and sodium hyaluronate were all dissolved in 0.1 M acetate buffer (pH 3.8 to 4.0) and used in the test.

Using the control blank CB1 as the measurement blank, the absorbances of the sample S1, the control C1 and the sample blank SB1 at a wavelength of 585 nm were measured. From the measurement results, the hyaluronidase inhibitory activity rate (%) was calculated according to the following formula (A). The test was performed 3 times and the average was expressed as 1 data. The higher the value of the hyaluronidase inhibitory activity rate, the higher the hyaluronidase inhibitory activity.

Hyaluronidase inhibitory activity rate (%) =
[{C1- (S1-SB1)} / C1] × 100 ... (A)
[In the formula, reference numeral C1 indicates the absorbance of control C1, S1 indicates the absorbance of sample S1, and SB1 indicates the absorbance of sample blank SB1. ]

Reference example 1
In Yamakawa-machi, Miyama City, Fukuoka Prefecture, 1.0 kg of white turmeric rhizomes harvested from November to December are washed with water, sliced to a thickness of 0 to 5 cm, dried at 60 to 70 ° C for 2 days, and residual water content. Was set to 10%.

Then, the dried slices of white turmeric were crushed using a screw type crusher and supervised to obtain dried turmeric powder having a particle size of 0.29 to 1.08 mm.

25 g of this powder was suspended in 50 ml of hexane and extracted at room temperature for 1 day. Then, it was filtered to remove the solid matter, and a hexane extract was obtained.

This hexane extract was analyzed using GC-MS (E1,70e, manufactured by Nippon Denshi Co., Ltd., JMN-ECA-600 type), and the peak area was compared with a pre-measured calibration curve to contain a labdane-type diterpene. When the total amount of similar compounds was determined, 45 μg of labdane-type diterpenes compound was contained in 1 ml of the extract.

Further, the hexane extract was fractionated by silica gel column chromatography using hexane, ethyl acetate, and methanol, and the Labdane-type diterpenes compound in the obtained fraction was measured for MS (mass spectrum) and 13C-NMR spectrum. As a result of confirmation, the spectrum data shown in Tables 1 to 4 were obtained.

Since the spectral data in Table 1 is consistent with the spectral data described in the literature (Bioscii. Biotechnol. Biochem., 70 (10), 2494-2500, 2006), the first component having the above spectral data is (E). ) -8β (17) -labd-12-en-15,16,17-trial.

Since the spectral data in Table 2 is consistent with the spectral data described in the literature (Bioscii. Biotechnol. Biochem., 66 (12), 2698-2700, 2002), the second component having the above spectral data is (E). ) -8β (17) -epoxylabd-12-en-15,16-jiar was identified.

Since the spectral data in Table 3 is consistent with the spectral data described in the literature (J. Nat. Prod., 60, 1287-1293, 1997), the third component having the above spectral data is 16-oxo-8. It was identified as (17), 12 (E) -labdaziene-15-euic acid.

Since the spectral data in Table 4 is consistent with the spectral data described in the literature (Phytochemistry, 36 (3), 699-701, 1994), the fourth component having the above spectral data is referred to as Rabda-8 (17). It was identified as 12-diene-15,16-jiar.

Example 1
A socksley extractor (SPC fat extractor) at 40 ° C. using a mixture of 25 g of white turmeric powder used in Reference Example 1 and 25 ml of hexane and 25 ml of rice oil (trade name: rice bran oil, manufactured by Oriza Yuka Co., Ltd.). , Shibata Kagaku Co., Ltd.), extracted continuously for 24 hours, and after extraction, solid-liquid separation was performed to obtain a hexane-rice oil mixed extract. The color of the obtained extract was measured as a blank of an equal amount mixture of hexane and rice oil, and the hyaluronidase inhibitory activity was measured. The results are shown in Table 5.

Example 2
In Example 1, extraction was carried out in the same manner as in Example 1 except that the extraction temperature was set to 100 ° C., and the appearance and hyaluronidase activity of the hexane-rice oil mixed extract were measured. The results are shown in Table 5.

Comparative Example 1
The same procedure as in Example 1 was carried out except that 25 ml of hexane was used instead of the mixture of 25 ml of hexane and 25 ml of rice oil in Example 1, and the appearance and hyaluronidase activity were measured using hexane as a blank. The results are shown in Table 5.

Comparative Example 2
The same procedure as in Example 2 was carried out except that 25 ml of hexane was used instead of the mixture of 25 ml of hexane and 25 ml of rice oil in Example 2, and the appearance and hyaluronidase activity were measured using hexane as a blank. The results are shown in Table 5.

As shown in Table 5, in both Examples 1 and 2 as compared with Comparative Examples 1 and 2, the results showed that the coloring was less (the absorbance was smaller) and the hyaluronidase inhibitory activity rate was higher, and rice bran oil, which is a vegetable oil, was obtained. The effect of preventing quality deterioration on labdane-type diterpenes compounds is clear.

Further, in Table 5, the hyaluronidase inhibitory activity of Examples 1 and 2 is at a high level, so it is clear that the labdane compounds, which are the active ingredients, are not resinified.

Example 3
40 ml of the hexane extract obtained in the same manner as in Reference Example 1 was divided into four equal parts, 10 ml of rice oil was added to the 10 ml, and the mixture was concentrated under reduced pressure in a bath at 40 ° C. using a rotary evaporator. Concentration was carried out until no hexane distilled. The hyaluronidase inhibitory activity of the obtained concentrated residue was measured, and the appearance of the concentrated residue was also measured. The results are shown in Table 6.

Example 4
In Example 3, 10 ml of the hexane extract divided into four equal parts was used, and the concentration was carried out in the same manner as in Example 3 except that the concentration was carried out under reduced pressure in a bath at 100 ° C., and the obtained concentrated residue was subjected to hyaluronidase inhibitory activity. The measurement was performed, and the appearance of the concentrated residue was measured by absorbance. The results are shown in Table 6.

Comparative Example 3
10 ml of the hexane extract divided into four equal parts in Example 3 was concentrated under reduced pressure in a bath at 40 ° C. in the same manner as in Example 3, and the hyaluronidase inhibitory activity of the obtained concentrated residue was measured. Measured in. The results are shown in Table 6.

Comparative Example 4
10 ml of the hexane extract divided into four equal parts in Example 3 was concentrated under reduced pressure in a bath at 100 ° C. in the same manner as in Example 4, and the hyaluronidase inhibitory activity of the obtained concentrated residue was measured. Measured in. The results are shown in Table 6.

As shown in Table 6, when rice oil was added and concentrated under reduced pressure, the hexane extract was concentrated as it was in Examples 3 and 4 as well, with less coloring (low absorbance) and hyaluronidase inhibitory activity. It did not decrease compared to the case of.

As is clear from Table 6, the concentrated residue of Comparative Example 3 in which the hexane extract was concentrated under reduced pressure in a bath at 40 ° C. or 100 ° C. was clearly colored. Furthermore, even if rice oil was added to the concentrated residue, it did not dissolve. Moreover, the concentrated residue of Comparative Example 4 was completely insoluble in hexane and rice oil.

From this result, it is clear that the addition of rice oil has an effect of preventing coloration and quality deterioration of the labdane-type diterpenes compound.

Example 5
A mixed solution of 25 ml of hexane and 25 ml of rice oil was added to 25 g of the white turmeric powder used in Reference Example 1 and left at 40 ° C. for 24 hours to extract a labdane-type diterpene compound. Then, solid-liquid separation was performed, and hexane in the extract was completely distilled off at 40 ° C. using a rotary evaporator.

Then, the content of the labdane-type diterpenes compound in the obtained concentrated residue was measured. The results are shown in Table 7.

Example 6
A mixed solution of 35 ml of hexane and 15 ml of rice oil was added to 25 g of the white turmeric powder used in Reference Example 1 and left at 40 ° C. for 24 hours to extract a labdane-type diterpene compound. Then, solid-liquid separation was performed, and hexane in the extract was completely distilled off at 40 ° C. using a rotary evaporator.

Then, the content of the labdane-type diterpenes compound in the obtained concentrated residue was measured. The results are shown in Table 7.

Comparative Example 5
In Example 5, 50 ml of hexane was used instead of the mixture of hexane and rice oil, and the same procedure as in Example 3 was carried out. The obtained concentrated residue was dissolved in 50 ml of DMSO, and labdane-type diterpenes in the solution were used. The content of the compound was measured. The results are shown in Table 7.

As shown in Table 7, in Examples 5 and 6 extracted by mixing rice oil with hexane, the content of the labdane-type diterpene compound was high, and when hexane was distilled off from the mixed solution, the labdane-type diterpene compound was easily obtained. It was confirmed that it was transferred to rice oil and concentrated.

Example 7
Using a mixture of 25 ml of ethyl alcohol and 25 ml of rice oil in 25 g of white turmeric powder used in Reference Example 1, continuous extraction was performed at 40 ° C. using a Soxhlet extractor for 24 hours, and after extraction, solid-liquid separation was performed. , Hexane-rice bran oil mixed extract was obtained. The obtained extract was concentrated under reduced pressure to remove ethyl alcohol, and the hyaluronidase inhibitory activity of the obtained concentrated residue was measured. The appearance of the concentrated residue was measured by absorbance. The results are shown in Table 8.

Example 8
In Example 7, the same procedure as in Example 7 was carried out except that the extraction temperature was 100 ° C., and the hyaluronidase inhibitory activity of the obtained concentrated residue was measured, and the appearance of the concentrated residue was measured by absorbance. The results are shown in Table 8.

Comparative Example 6
In Example 7, the same procedure as in Example 7 was carried out except that 25 ml of ethyl alcohol was used instead of the mixture of 25 ml of ethyl alcohol and 25 ml of rice oil, and the hyaluronidase inhibitory activity was measured for the obtained concentrated residue. The appearance of the concentrated residue was measured by absorbance. The results are shown in Table 8.

Comparative Example 7
In Example 8, the same procedure as in Example 8 was carried out except that 25 ml of ethyl alcohol was used instead of the mixture of 25 ml of ethyl alcohol and 25 ml of rice oil, and the hyaluronidase inhibitory activity was measured for the obtained concentrated residue. The appearance of the concentrated residue was measured by absorbance. The results are shown in Table 8.

As shown in Table 8, it was obtained that Example 7 had less coloring (lower absorbance) and a higher hyaluronidase inhibitory activity rate than Comparative Example 6, and Example 8 had a higher hyaluronidase inhibitory activity rate than Comparative Example 7. The results show that there is little coloring and the hyaluronidase inhibitory activity rate is high, and the effect of rice bran oil, which is a vegetable oil, on labdane-type diterpenes compounds is clear.

The present invention can be implemented in various other forms without departing from its spirit or key features. Therefore, the above-described embodiment is merely an example in all respects, and the scope of the present invention is shown in the claims and is not bound by the text of the specification. Furthermore, all modifications and modifications that fall within the scope of the claims are within the scope of the present invention.

Claims (8)

A stabilizing preparation for oral administration, characterized in that a labdane-type diterpene compound is contained in vegetable oil. The oral according to claim 1, wherein the labdane-type diterpene compound is extracted from a plant containing the labdane-type diterpene compound by an organic solvent, and the vegetable oil contains substantially no organic solvent. Stabilizing preparation for administration. The labdane-type diterpenes compounds are (E) -8β (17) -labd-12-ene-15,16,17-trial, (E) -8β (17) -epoxy labdone-12-ene-15,16-. It is characterized by being one or more selected from diterpene, 16-oxo-8 (17), 12 (E) -labdane diene-15-oic acid and labdane-8 (17), 12-diene-15,16-dier. The stabilizing preparation for oral administration according to claim 1 or 2. The stabilized preparation for oral administration according to any one of claims 1 to 3, wherein the vegetable oil is rice bran oil. (A) A plant containing a labdane-type diterpene compound is extracted with a sex organic solvent, and then vegetable oil is added to the obtained extract to prepare an extraction mixture, or (B) a plant containing a labdane-type diterpene compound. Is extracted with a mixture of an organic solvent and a vegetable oil to obtain an extraction mixture, and the organic solvent is removed from the obtained extraction mixture to obtain a stabilized preparation for oral administration in which a labdane-type diterpene compound is contained in the vegetable oil. A method for producing a stabilizing preparation for oral administration. Turmeric is a plant containing labdane-type diterpenes compounds.
The method for producing a stabilized preparation for oral administration according to claim 5, wherein the organic solvent is at least one selected from ethyl alcohol, methyl alcohol, acetone, hexane, cyclohexane, heptane and diethyl ether. After extracting a plant containing (C) a labdan-type diterpene compound with an organic solvent, vegetable oil is added to the obtained extract to prepare an extraction mixture, or (D) a plant containing a labdan-type diterpene compound is used. , A method for concentrating a Labdan-type diterpene compound, which comprises extracting with a mixture of an organic solvent and a vegetable oil to prepare an extraction mixture, concentrating the obtained extraction mixture, and removing the organic solvent. A composition for oral ingestion containing a labdane-type diterpene compound, which comprises a labdane-type diterpene compound contained in a vegetable oil.