JP6715007B2 - Natural benzaldehyde water production method, natural benzaldehyde water produced by the production method, and antitumor agent containing the natural benzaldehyde water - Google Patents

Description

The present invention relates to a method for producing natural benzaldehyde water, natural benzaldehyde water produced by the production method, and an antitumor agent containing the natural benzaldehyde water.

Extracts from plants belonging to the genus Rosaceae, such as sakura and plum, have antioxidative action (Patent Document 1), anti-inflammatory action (Non-Patent Document 1), nitric oxide production inhibitory action (Patent Document 2), melanin. It is already known to have a production suppressing action (Patent Document 3), a whitening action (Non-Patent Document 2), a moisturizing action (Patent Document 4) and the like.

Further, Patent Document 5 and Non-Patent Document 3 disclose that an extract from a plant belonging to the genus Sakura of the Rosaceae family contains benzaldehyde, and benzaldehyde has an antitumor effect (non-patent document). It is already known that there is a blood flow promoting action (Patent Document 6) and the like.

However, an extraction method for extracting more components having an effective action as described above from a plant belonging to the genus Rosaceae, and the production of an extract containing more benzaldehyde and trace components other than benzaldehyde from the plant. No way is known so far.

On the other hand, as a method of extracting and separating components contained in the plant from the plant, a method of separating the upper layer (oil phase) separated into two layers by extracting using a steam distillation method; heating by directly immersing in water The direct extraction method of separating the upper layer (oil phase) from the two separated layers; the solvent extraction method of extracting with an organic solvent; the compression method of extracting by adding an oily component and pressing; the supercritical fluid Various methods are known, such as a supercritical extraction method in which the extraction is performed using the method.
However, these extraction methods do not separate the aqueous phase as the main target substance, and are particularly unsuitable as a method for separating the aqueous phase containing benzaldehyde which is slightly soluble in water.

Further, a low temperature vacuum extraction method is known in which an extraction solvent such as an organic solvent, water or water vapor is not used and the pressure is reduced at a low temperature to directly extract.
Patent Document 7 describes a method for extracting an aroma component in which a plant-derived vapor is generated by heating and decompressing a plant while crushing and stirring the plant. It is said that the aroma component can be extracted from herbs, fruits, flowers, or vegetables by using this method.

However, as described above, components contained in plants belonging to the genus Rosaceae, up to now, extraction methods to extract more or more efficiently from the plant, benzaldehyde from the plant, components other than benzaldehyde coexisting with it or the It does not disclose a method for producing an extract containing a large amount of plant cell water.
Furthermore, an extraction method for efficiently extracting a plurality of active ingredients containing unidentified molecules as an aqueous phase from a plant belonging to the genus Sakura of the family Rosaceae, and the production of the extract as it contains cell water of the plant The method is not disclosed either.

JP, 2006-166726, A JP, 2012-229170, A JP, 2011-016727, A JP, 2002-020225, A JP, 2001-252053, A JP, 2000-169326, A Japanese Patent Laid-Open No. 2012-062374

Lee J., BMC Complement Altern Med., 2013 Apr;13:92 Murata K., Nat Prod Commun., 2014 Feb;9(2):185-8 Ulker Z., Hum Exp Toxicol., 2013 Aug;32(8):858-64. Kochi M., Cancer Treat Rep., 1980 Jan;64(1):21-3

The present invention has been made in view of the above background art, and its problem is to solve the above problems, to provide a method for producing an aqueous plant extract containing a large amount of benzaldehyde existing in nature, and The present invention provides a benzaldehyde produced by the production method and a plant extract containing a large amount of components coexisting with benzaldehyde in plant cells.

The present inventor has conducted extensive studies to solve the above problems, and by using a specific extraction method, as an aqueous phase, contains a large amount of benzaldehyde present in "plants belonging to the genus Sakura of the family Rosaceae". It was found that an extract can be obtained. Further, they have found that, in addition to benzaldehyde, the extract contains a large amount of a plurality of active ingredients including cell water without being volatilized, and completed the present invention.
Further, they have found that the extract has an excellent antitumor effect, and completed the present invention.

That is, the present invention is a method for producing natural benzaldehyde water, in which a plant belonging to the genus Prunus genus Prunus genus is crushed and stirred with a stirring blade, and the crushing and stirring is performed while applying heat from the outside to reduce pressure for extraction. A method for producing natural benzaldehyde water, which comprises an extraction method having steps.
Hereinafter, the "extraction method having a step of stirring while crushing with a stirring blade and depressurizing while applying heat from the outside while crushing and stirring" is simply referred to as "low temperature vacuum extraction method".

The present invention also provides a natural benzaldehyde water produced by the above-mentioned method for producing a natural benzaldehyde water.

The present invention also provides an antitumor agent, which comprises the above-mentioned natural benzaldehyde water.

According to the present invention, the above-mentioned problems and the above-mentioned problems are solved, and it was difficult to obtain by other conventional extraction methods, and an extract derived from a plant belonging to the genus Rosaceae, which contains a large amount of benzaldehyde in the cells. It is possible to obtain
Also, for components other than benzaldehyde, it is possible to obtain an extract from a plant of the genus Prunus rosacea with "components" or "component ratios" that were difficult to obtain efficiently by conventional extraction methods. is there. Moreover, since the thermal decomposition can be prevented, the natural component can be directly used as the extract.

Further, according to the present invention, a large amount of extract liquid, which cannot be obtained by other conventional extraction methods, can be obtained as an aqueous phase.
In the production method of the present invention, since it is not necessary to use an extraction solvent such as water or an organic solvent, a plant-derived component (including cell water) can be recovered as an aqueous solution at a high concentration in good yield, Since the obtained extract can contain no substance derived from the extraction solvent, it can be composed of only naturally-derived components including water, is extremely safe, and can be provided with peace of mind. No extraction solvent odor.

In addition, by having a “component” or “component ratio” that was not present in the extract of the plant belonging to the genus Rosaceae, a high-quality and novel extract derived from the plant belonging to the genus Rosaceae Can be provided.

Further, the natural benzaldehyde water of the present invention has an excellent antitumor effect.
The "natural benzaldehyde water", which is an extract derived from a plant belonging to the genus Prunus of the present invention, has excellent superoxide anion radical scavenging activity, singlet oxygen scavenging activity, hydroxy radical scavenging activity, nitric oxide scavenging activity. It is active and can suppress the amount of the stress hormone cortisol.

It is the schematic which shows one form of the apparatus used for the extraction process in this invention. It is an expanded sectional view showing one form of a container of a device used for an extraction process in the present invention. It is a perspective view which shows one form of the structure of the stirring blade in the container of the apparatus used for the extraction process in this invention. It is a graph which shows the component and component ratio of the natural benzaldehyde water extracted from the flower of a cherry tree. It is a graph which shows the component and component ratio of the natural benzaldehyde water extracted from the cherry tree leaf. It is a graph which shows the result of having measured the superoxide anion radical scavenging activity of the natural benzaldehyde water of this invention. It is a graph which shows the result of having measured the singlet oxygen scavenging activity of the natural benzaldehyde water of this invention. It is a graph which shows the result of having measured the hydroxyl radical scavenging activity of the natural benzaldehyde water of this invention. It is a graph which shows the result of having measured the nitric oxide scavenging activity of the natural benzaldehyde water of this invention. Measurement of changes in saliva volume (μL) (A), saliva pH (B), and superoxide anion radical scavenging activity (%) in saliva when the subject natural benzaldehyde water of the present invention was vaporized and smelled. It is a graph which shows the result. Changes in the antioxidant capacity of saliva (μmol/L) (A) and the amount of cortisol in saliva (μg/dL) (B) were measured when the natural benzaldehyde water of the present invention was vaporized and allowed to smell. It is a graph which shows a result. It is a graph which shows the result of having measured the antitumor activity of the natural benzaldehyde water of this invention using A549 cell. It is a graph which shows the result of having measured the antitumor activity of the natural benzaldehyde water of this invention using A431 cell. It is a graph which shows the result of having measured the antitumor activity of the natural benzaldehyde water of this invention using Hela cells.

Hereinafter, the present invention will be described, but the present invention is not limited to the following specific forms and can be arbitrarily modified within the scope of the technical idea.

The present invention is a method for producing natural benzaldehyde water, in which a plant belonging to the genus Rosaceae is crushed and stirred with a stirring blade, and a step of decompressing while externally applying heat to the crushing/stirring is performed. A method for producing natural benzaldehyde water, comprising a step of extracting by an extraction method having.

In the present specification, "natural benzaldehyde" is defined as "an organic component which is mainly composed of benzaldehyde extracted from a plant cell and coexists in the plant cell with the benzaldehyde", and "natural benzaldehyde water" is the above. It is defined as "natural benzaldehyde" and an aqueous solution containing cell water of the above plant.
"Cell water" is the intracellular fluid contained in plant cells.

In the production method of the present invention, a plant belonging to the genus Prunus of the family Rosaceae (hereinafter sometimes simply abbreviated as “Prunus plant”) is used as a raw material.
The plant belonging to the genus Sakura of the family Rosaceae is not particularly limited, but it is an excellent supermarket that it is one or more plants selected from the group consisting of cherry, plum, loquat, almond, peach, plum, prune and nectarine. Preferred from the viewpoints of oxide anion radical scavenging activity, singlet oxygen scavenging activity, hydroxy radical scavenging activity, nitric oxide scavenging activity, etc., and cherry is more preferred from the above-mentioned points, for example, Prunus yedoensis, Sekiyama ( Prunus yedoensis cv. Sekiyama) and the like. The double-flowered cherry tree is particularly preferable from the above-mentioned point and especially from the viewpoint that the number of petals is large or the volume of petals is large.

The plant of the genus Prunus may be extracted from any tissue of the plant of the genus Prunus, and leaves of the plant of the genus Prunus; stem; corolla of flower petals; calyx; fruit; seed; kernel; tree such as bark; root etc. However, it is preferably extracted from petals, leaves or kernels, more preferably extracted from petals or leaves, and extracted from petals, in that natural benzaldehyde water containing a large amount of natural benzaldehyde can be obtained. Is particularly preferable. In addition, when extracting from ume, it is particularly preferable to extract from jin from the above point.
Moreover, you may extract simultaneously from the above-mentioned several tissues of the plant which belongs to Rosaceae genus Sakura.

What is used for extraction may be either non-dried (raw) or dried to a certain extent, but preferably non-dried (substantially raw) contains cell water. Since it is preferable that the content of the benzaldehyde is sufficient, there is no deterioration, and that no extraction solvent including water is used in the production method of the present invention, in that case, natural benzaldehyde water consisting only of natural components can be suitably obtained as an aqueous solution. Is preferred.
When extracted from a non-dried product, it contains a large amount of cell water, so benzaldehyde and "components that coexist with benzaldehyde in plant cells" can be obtained in a completely dissolved form in a large amount of cell water, producing a uniform natural benzaldehyde water. it can.

The natural benzaldehyde water in the present invention is an extract extracted from a plant of the genus Prunus by a low temperature vacuum extraction method. In other words, the plant of the genus Prunus was extracted by the "extraction method including a step of stirring while crushing with a stirring blade and depressurizing while applying heat from the outside while crushing and stirring" (low temperature vacuum extraction method). It is an extract of the genus Prunus.
Here, the form of the genus Prunus plant to be subjected to the low-temperature vacuum extraction method, that is, the form before stirring while crushing the genus Prunus plant with stirring blades, easy to put into the device used in the low-temperature vacuum extraction method, in the device There is no particular limitation as long as it is a suitable form, but it is also preferable that it is appropriately cut.

When using petals, it is preferable to remove stamens, pistils, sepals, etc. before putting them in the apparatus. When using a leaf, a leaf area after ^cutting, preferably 0.2 cm 2 to 50 cm ^2, more preferably 1cm ² ~40cm ^2, particularly preferably 2cm ² ~30cm 2.

The method for producing natural benzaldehyde water of the present invention is a method for producing an extract extracted from a plant belonging to the genus Rosaceae (Prunus genus) using a low temperature vacuum extraction method.

In the low-temperature vacuum extraction method, extraction solvents such as organic solvent, water, water vapor, and liquid carbon dioxide are not substantially used, and direct extraction under reduced pressure at low temperature is not possible because the solvent cannot remain and only natural ingredients are extracted. Natural benzaldehyde water can be produced, no odor of solvent remains, content components do not decompose by heat, content components do not dissipate, high concentration of natural benzaldehyde water is obtained, and cell water does not dissipate, so a large amount of natural benzaldehyde water is obtained It is preferable in that it can be obtained.
Here, "low temperature" refers to a temperature lower than a general temperature at the time of solvent extraction without depressurization, and in the case of the present invention, specifically, (at the temperature of the extraction container as well as the gas in the extractor, It is preferable that the temperature of the genus Prunus itself is 90° C. or lower. The extraction temperature will be described in detail later.

<Crushing, stirring and decompressing while applying heat from the outside>
The method for producing natural benzaldehyde water according to the present invention has a step of extracting a plant of the genus Prunus by stirring with a stirring blade while crushing and depressurizing while applying heat from the outside while crushing and stirring.

<<Extractor>>
FIG. 1 is a schematic view showing one form of an apparatus used in an extraction step in the method for producing natural benzaldehyde water of the present invention. As long as it is within the scope of the present invention, it is not limited to the one extracted by the device shown in the drawing.
The container 1 is a container for accommodating the genus Prunus, stirring and crushing it with a stirring blade, and decompressing while applying heat from the outside while crushing and stirring, and the cooling tank 2 exits from the container 1. It is a device that cools steam.

The container 1 is composed of a lower semi-cylindrical portion 7 accommodating the stirring blade 6 and an upper rectangular portion 8 formed thereon. Around the lower semi-cylindrical portion 7 is a steam chamber 9 for applying heat to the inside of the container 1.
At the center of the lowermost part of the lower semi-cylindrical portion 7, there is provided an outlet 10 for taking out the crushed material of the genus Prunus after extraction.

An exhaust port 14 for the sucked vapor is provided in the upper part of the upper rectangular portion 8, and a pipe 16 connected to the cooling tank 2 is connected to the exhaust port 14.
At the upper part of the upper rectangular portion 8, an input port 17 for the genus Prunus is provided, and a lid 18 for closing the input port 17 is provided.

In the method for producing natural benzaldehyde water according to the present invention, it is essential that the plant of the genus Prunus is stirred while being crushed by a stirring blade, and extraction is performed under the crushing and stirring. In this way, by extracting, if a fresh crushed surface can be immediately extracted from it, it is possible to prevent thermal decomposition of the active ingredient, denaturation due to oxidation, etc., and put it in the extraction device of the present invention. It is possible to prevent the active ingredient from being dissipated by evaporation or the like from the crushed surface when it is crushed before being crushed.
It is particularly preferable to carry out the crushing/stirring in an extracting device equipped with a movable blade and/or a fixed blade in order to obtain the above-mentioned effects.

For example, FIG. 3 is a perspective view showing the structure of the stirring blade 6, which is rotated by a motor provided outside the container 1 and is attached to the end walls 20 and 21 of the container 1. Since the left and right end plates 22 and 23 are rotatably supported and the vanes 24 and 25 are fixed to both ends between the ends, the vanes 24 and 25 are substantially V-shaped, the central axis is not provided. The structure is organized.

Reference numeral 26 denotes a plurality of fixed blades fixed to the inner surface of the lower semi-cylindrical portion 7, and the portion of the blade bodies 24, 25 corresponding to the fixed blade 26 passes through the portion of the fixed blades 26 of the blade bodies 24, 25. Grooves 24a, 25a for forming are formed, and movable blades 24b, 25b for cutting the plant 31 belonging to the genus Rosaceae (Prunus plant 31) between the fixed blades 26 are formed on both sides of the groove 24a, 25a. ing.
Note that, in FIG. 3, the fixed blade 26 and the movable blades 24b and 25b are arranged so as to be displaced in the circumferential direction so that the meshing is sequentially performed with a time shift, whereby the drive motor of the stirring blade 6 is driven. We try to prevent momentary increase in power.

As shown in FIG. 2, an inclined surface 30 is provided at an upper portion on one side of the lower semi-cylindrical portion 7 so that a plant 31 belonging to the genus Rosaceae belonging to the genus Rosaceae can drop smoothly.
Reference numeral 32 is a vacuum gauge for measuring the degree of vacuum in the container 1, 33 and 34 are thermometers, which measure the pressure (pressure reduction degree) and temperature in the container in the extraction step, and the genus Prunus 31 at the time of extraction Is also provided for indirect measurement of the temperature, and is also provided for determining the start and end of extraction.

<<Extraction process>>
The operation/operation of this vacuum drying device is performed as follows, for example.
First, at the start of work, the cooling water is filled in the cooling tank 2. Then, the genus Prunus 31 is put into the container 1 through the inlet 17 and the lid 18 is closed. Then, the stirring blade 6 is rotated in the direction of the arrow R of FIGS. 1 to 3 to stir the genus Prunus in the container 1, while the genus Prunus 31 between the movable blades 24 b and 25 b and the fixed blade 26. Crush into small pieces.

By extracting while crushing the plant of the genus Prunus, it is possible to extract from a fresh crushed surface, and it is possible to prevent "denaturation" of the components, that is, natural benzaldehyde and cell water, and "loss by weight loss".

Simultaneously with the stirring and crushing, by supplying heating steam into the steam chamber 9, heat is applied from the outside. The heat applied to the container 1 is transferred to the cherry tree genus plant 31, and the cherry tree genus plant 31 is stirred by the stirring blades 6, whereby the extraction is promoted. This extraction is further promoted by the fact that the genus Prunus 31 is crushed by the movable blades 24b and 25b and the fixed blade 26 and becomes small.
At that time, the temperature and amount of the heating steam fed into the steam chamber 9 are adjusted to bring the temperature of the genus Prunus 31 itself into a preferable range described later.

By sucking with the decompression device 46 such as an ejector or a vacuum pump, the gas in the container 1, that is, the vapor and air of the extract is sucked through the pipe 16 and contained in the genus Prunus 31 in the container 1. Evaporation of natural benzaldehyde and cell water begins.
At that time, the amount of suction and the suction force by the depressurizing device 46 are adjusted so that the pressure (degree of depressurization) at the time of extraction falls within a preferable range described later.

The “steam of natural benzaldehyde contained in the plant of the genus Prunus” and “steam of water which is the main component of cell water” (steam) in the container 1 are sucked through the pipe 16 and introduced into the cooling tank 2 and liquefied. Then, it becomes a recovery liquid and accumulates in the recovery tank 41.

When the collection liquid (oil layer 50 and water layer 51) has accumulated up to a predetermined amount in the collection tank 41, suction by the decompression device 46 is stopped, the valve 45 is closed, and the valve 49 is opened to collect the collection liquid. If necessary, the recovered liquid is allowed to stand still for liquid separation to remove the oil layer 50 and recover the aqueous layer 51 as an extract (natural benzaldehyde water).

In the present invention, containing natural cell water, containing only natural benzaldehyde as an active ingredient, containing a plurality of medicinal ingredients other than benzaldehyde, natural benzaldehyde is uniformly dissolved in cell water Widely used for water-based applications, especially suitable for atomization using a diffuser (fragrance diffuser), which is a suitable device for nasal administration (high diffuser compatibility), Prunus It is particularly preferable to use the aqueous layer (aqueous phase, aqueous fraction) because the aqueous layer is mainly collected from the petals of the plant.

<<Extraction conditions>>
In the method for producing an extract of the present invention, the temperature of the extraction is not particularly limited, but it is preferable that the genus Prunus itself is 90° C. or lower, and it is more preferable that the temperature is 55° C. or lower. .. Particularly preferably, the extraction is performed so that the genus Prunus itself maintains a temperature of 20°C or higher and 50°C or lower, more preferably 25°C or higher and 45°C or lower, and most preferably 30°C or higher. It is 40°C or lower.

When the lower limit is a value within the above range, thermal decomposition does not occur and the extraction time can be shortened, so decomposition and loss of the active ingredient are suppressed, and unnecessary time is not lost, which is economical. In addition, the active ingredients (natural benzaldehyde and cell water) can be sufficiently extracted.
In the method for producing natural benzaldehyde water of the present invention, the genus Prunus is stirred while being crushed by a stirring blade, and extraction is performed under the crushing/stirring. However, if the lower limit of the temperature is within the above range, the extraction time can be shortened, and the effect is further synergized.
On the other hand, when the upper limit is a value within the above range, the active ingredients (natural benzaldehyde and cell water) can be extracted while preventing denaturation/decomposition of the active ingredient due to heat. In addition, unnecessary components are not extracted.

In the method for producing an extract according to the present invention, the degree of reduced pressure in the above extraction is not particularly limited, but it is preferable to perform the extraction while maintaining a pressure lower than 801.3 kPa (1 atm). The pressure is more preferably 85 kPa or more lower than 101.3 kPa (1 atm), particularly preferably 90 kPa or more lower pressure, and more preferably 95 kPa or more lower pressure.

When the pressure is the above value (the degree of pressure reduction is the above), it is possible to extract the active ingredient such as natural benzaldehyde at a low temperature, so that it is possible to prevent the active ingredient from being denatured and decomposed by heat, and it is sufficiently effective. The ingredients can be extracted. Further, since the extraction time can be shortened, decomposition and loss of the active ingredient are suppressed, and there is no unnecessary time loss, which is economical. In addition, natural benzaldehyde and cell water can be sufficiently extracted.
In the method for producing an extract of the present invention, the genus Prunus is agitated while being crushed by a stirring blade, and the extraction is performed under the crushing/agitation, so it may be extracted early when a fresh crushed surface is formed. However, when the degree of reduced pressure is low as in the above value, the effect of suppressing the decomposition and loss of the active ingredient such as natural benzaldehyde is further synergized in that the extraction rate is high.

<Comparison with other extraction methods>
As described above, as the extraction method, in addition to the low temperature vacuum extraction method of the present invention, various methods such as a steam distillation method, a direct extraction method, a solvent extraction method, a squeezing method and a supercritical extraction method are known.

Among them, the steam distillation method and the direct extraction method cannot sufficiently obtain an extract containing natural benzaldehyde, because the active ingredient is denatured and the active ingredient is dissipated because the plant of the genus Prunus is heated at a high temperature.
Further, in the steam distillation method, a liquid (that is, natural benzaldehyde water) that also contains benzaldehyde and trace components that coexist in plant cells (benzyl alcohol, coumarin, and other components of minute peaks shown in FIGS. 4 and 5). ) Cannot be obtained. Further, in the steam distillation method, the effect of the present invention that all of the natural benzaldehyde water of natural origin is eliminated by mixing the liquefied water of the steam used for extraction with the cell water.

In addition, in the solvent extraction method, it is difficult to extract the water-soluble component, the extraction solvent remains in the extraction liquid, and the active component is also removed when the solvent is removed. Further, in the solvent extraction method, since it is essential to use a solvent, it was difficult to obtain plant-derived high-concentration cell water. Furthermore, the yield of the extract (natural benzaldehyde water) was also low. Even when water is used as the extraction solvent, the water added for extraction remains, so that the effect of the present invention that the water is naturally derived natural benzaldehyde disappears.

Even by the pressing method, the oily component used as the extraction solvent remained in the extract and it was impossible to completely remove the oily component. Moreover, since an extraction solvent is used, it is difficult to obtain plant-derived high-concentration cell water.
The supercritical extraction method has a problem in that it requires high pressure and thus requires expensive equipment.

In addition, a method in which the genus Prunus genus is not crushed and stirred but is extracted once after crushed and stirred sometimes cannot efficiently extract a plurality of active ingredients such as natural benzaldehyde. In a method of extracting by heating and decompressing instead of crushing and stirring as in the usual vacuum extraction method, etc., active ingredients such as natural benzaldehyde (particularly benzyl alcohol, coumarin, and others, as shown in FIGS. In some cases, it was not possible to extract the components of the minute peaks shown).

In addition, in other extraction methods including the usual vacuum extraction method in which the genus Prunus plant is heated and decompressed "without being crushed", various other types of genus contained in the tissues and cells of the genus Prunus plant are included. There was a case where the component of was not efficiently and sufficiently extracted.

<Operation after extraction>
According to the low temperature vacuum extraction method of the present invention, uniform natural benzaldehyde water is obtained from the aqueous fraction (aqueous layer, aqueous phase) and essential oil is obtained from the oily fraction (oil layer, oil phase).
In the present invention, the operation after extraction is not particularly limited, but it is preferable to separate and remove unnecessary components that are recovered at the same time. For the separation, decantation, liquid separation operation, etc., which utilizes the difference in specific gravity, etc. are used.
The present invention is characterized in that the extract can be obtained as "natural benzaldehyde water" containing no non-plant-derived component.

If it is natural benzaldehyde water, it can be easily sprayed indoors using a general-purpose sprayer (for example, a humidifier etc.) and the filter etc. does not get dirty, so frequent maintenance such as filter replacement is required. Has the effect of not. Further, the effect of containing a large amount of active ingredients other than benzaldehyde can be obtained.
In addition, a larger amount of the water-soluble extract can be recovered in one extraction step than the oil-soluble extract. Further, although the odor is lighter than that of the oil-soluble extract, it has a odor intensity of a level that is relatively comfortable, and is easy to use in the medical field or at home. Further, it is superior to the oil-soluble extract in that it is easy to wash containers and parts.

The natural benzaldehyde water obtained by the “method for producing natural benzaldehyde water” of the present invention has a different peak (ratio) in chromatograph as compared with the extract from the conventional plant of the genus Prunus, Is a new extract.
Such natural benzaldehyde water of the present invention contains benzaldehyde, benzyl alcohol, and coumarin as shown in FIGS. 4 and 5 in addition to water, and other components of the minute peaks shown in FIGS. In addition, it also contains trace components that cannot be detected by chromatography.
The extract from the conventional plant of the genus Prunus extracted by an extraction method other than the low-temperature vacuum extraction method contains benzaldehyde, but does not contain the above-mentioned other components or contains only a trace amount, or the content ratio. Was very different. Further, it contained a decomposition product of benzaldehyde and an extraction solvent at the time of extraction.
Since the difference in the component composition and the physical property value has not been clarified, the natural benzaldehyde water of the present invention can be specified only by the production method.

The natural benzaldehyde water of the present invention exhibits the following effects as compared with benzaldehyde.

The natural benzaldehyde water of the present invention has an antioxidant effect. In addition, it has a scavenging activity of superoxide anion radicals (O ₂ ^·− ), singlet oxygen ( ¹ O ₂ ), and hydroxy radicals (HO ^· ), which are types of active oxygen. In particular, it has an excellent scavenging activity against superoxide anion radicals and hydroxy radicals.
Natural benzaldehyde water itself, food and drink containing the natural benzaldehyde, or by evaporating the natural benzaldehyde water into the body, the active oxygen is eliminated, and the antioxidant action and anti-aging (anti-aging) effect Can be demonstrated.

Moreover, the natural benzaldehyde water of the present invention has an anti-inflammatory effect. It also has a nitric oxide (NO) erasing activity.

The natural benzaldehyde water of the present invention may be used as a solution by diluting it with water, an organic solvent or the like, if necessary, and if necessary, various additives may be added as a drug. Can be used in any form. However, the natural benzaldehyde water of the present invention is used as it is (or in addition to natural ingredients), for example, in various applications described below in order to take advantage of the fact that it is made of natural products including water. Is also preferable.

The natural benzaldehyde water of the present invention is of plant origin and does not require the use of an extraction solvent in the extraction step, so it is extremely safe and can be used with confidence.
In addition, the natural benzaldehyde water of the present invention can be used for medicines, quasi drugs, vaporizing suction agents, external compositions, perfumes, foods and drinks, cosmetics, bath agents, fibers and the like. When used for these purposes, various additives can be mixed therein, if necessary.

In addition, when the natural benzaldehyde water of the present invention or an agent containing the natural benzaldehyde water was actually sniffed by a human, the antioxidant capacity in saliva was significantly improved 30 minutes after use, The natural benzaldehyde water of the invention and the method having a step of smelling the agent containing the natural aldehyde water are particularly effective as a method for improving the antioxidant ability in saliva.

Further, when the human was actually sniffed with the natural benzaldehyde water of the present invention or the agent containing the natural benzaldehyde water, cortisol, which is a stress hormone in saliva of the human, was significantly increased 30 minutes after use. Since it has decreased, the method of the present invention comprising the step of smelling the natural benzaldehyde water and the agent containing the natural aldehyde water is effective as a method for suppressing the content of cortisol in saliva.

<Antineoplastic agent>
The antitumor agent of the present invention is characterized by containing the above-mentioned natural benzaldehyde water.
The superior antitumor effect was confirmed in human alveolar basal epithelial adenocarcinoma cell A549, human squamous cell carcinoma cell A431, and human cervical cancer cell HeLa. The antitumor agent of the present invention can be used for prevention of tumor development and/or treatment of tumor.

Further, the antitumor agent of the present invention can contain "other components" in addition to natural benzaldehyde water, which is an active component.

The "other component" in the antitumor agent is not particularly limited and may be appropriately selected depending on the purpose within a range not impairing the effects of the present invention, and for example, pharmaceutically acceptable A carrier etc. are mentioned.
The carrier is not particularly limited and can be appropriately selected depending on, for example, the dosage form described below. The content of the "other components" in the antitumor agent is also not particularly limited and can be appropriately selected according to the purpose.

The dosage form of the antitumor agent of the present invention is not particularly limited and can be appropriately selected depending on, for example, a desired administration method described below.
Specifically, for example, oral solid preparations (tablets, coated tablets, granules, powders, capsules, etc.), oral liquid preparations (oral liquid preparations, syrups, elixirs, etc.), injections (solvents, suspensions, etc.) , Ointments, patches, gels, creams, external powders, sprays, inhalation sprays and the like.

As the oral solid preparation, for example, an excipient, and if necessary, an additive such as a binder, a disintegrating agent, a lubricant, a coloring agent, and a flavoring/flavoring agent are added to the above-mentioned active ingredient, and a conventional method is used. Can be manufactured by.
Examples of the excipient include lactose, sucrose, sodium chloride, glucose, starch, calcium carbonate, kaolin, microcrystalline cellulose, silicic acid and the like.
Examples of the binder include water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl starch, methyl cellulose, ethyl cellulose, shellac, calcium phosphate, polyvinylpyrrolidone and the like. To be
Examples of the disintegrant include dry starch, sodium alginate, agar powder, sodium hydrogen carbonate, calcium carbonate, sodium lauryl sulfate, stearic acid monoglyceride, lactose and the like.
Examples of the lubricant include refined talc, stearate, borax, polyethylene glycol and the like.
Examples of the colorant include titanium oxide and iron oxide.
Examples of the corrigent/flavoring agent include sucrose, orange peel, citric acid, tartaric acid, and the like.

The oral liquid preparation can be produced, for example, by a conventional method by adding additives such as a flavoring/flavoring agent, a buffering agent, and a stabilizer to the active ingredient.
Examples of the corrigent/flavoring agent include sucrose, orange peel, citric acid, tartaric acid and the like. Examples of the buffer include sodium citrate and the like. Examples of the stabilizer include tragacanth, gum arabic, gelatin and the like.

The injection includes, for example, subcutaneously, intramuscularly, and intravenously by adding a pH adjusting agent, a buffering agent, a stabilizer, an isotonicity agent, a local anesthetic, etc. to the above-mentioned active ingredient. And the like can be produced.
Examples of the pH adjuster and the buffer include sodium citrate, sodium acetate, sodium phosphate and the like. Examples of the stabilizer include sodium pyrosulfite, EDTA, thioglycolic acid, thiolactic acid and the like. Examples of the tonicity agent include sodium chloride, glucose and the like. Examples of the local anesthetic include procaine hydrochloride, lidocaine hydrochloride and the like.

The ointment can be produced, for example, by mixing a known base, a stabilizer, a wetting agent, a preservative and the like with the active ingredient and mixing them by a conventional method.
Examples of the base include liquid paraffin, white petrolatum, white beeswax, octyldodecyl alcohol, paraffin and the like. Examples of the preservative include methyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate and the like.

The patch can be produced, for example, by applying a cream, gel, paste or the like as the ointment to a known support by a conventional method. Examples of the support include woven fabrics, non-woven fabrics made of cotton, staple fibers and chemical fibers, films of soft vinyl chloride, polyethylene, polyurethane and the like, foam sheets and the like.

The content of natural benzaldehyde water, which is the active ingredient in the antitumor agent of the present invention, with respect to the whole antitumor agent is not particularly limited and can be appropriately selected according to the purpose. When the total amount of natural benzaldehyde water is 0.001 to 100 parts by mass, the content is preferably 0.001 to 100 parts by mass, more preferably 0.01 to 99 parts by mass, and particularly preferably 0.1 to 95 parts by mass. It may be added in an amount of 1 part by mass, more preferably 1 to 90 parts by mass.

The animal to which the antitumor agent of the present invention is administered is not particularly limited, and examples thereof include humans; mice; rats; monkeys; horses; livestock such as cows, pigs, goats, chickens; pets such as cats and dogs; Are listed.

Further, the administration method of the antitumor agent is not particularly limited, for example, depending on the dosage form of the antitumor agent, it can be appropriately selected, oral administration, intraperitoneal administration, respiratory inhalation, Examples include injection into blood and intestinal injection.
The dose of the antitumor agent is not particularly limited and may be appropriately selected depending on the age, body weight, desired degree of effect, etc. of the subject to be administered. The daily dose is preferably 1 mg to 30 g, more preferably 10 mg to 10 g, and particularly preferably 100 mg to 3 g as the amount of the active ingredient.
The administration timing of the antitumor agent is also not particularly limited and may be appropriately selected depending on the purpose. For example, it may be administered prophylactically or therapeutically.

<Action/Principle>
In the present invention, the action/principle of the extract of the genus Prunus plant (natural benzaldehyde water) extracted by the low-temperature vacuum extraction method having an excellent antitumor effect is not clear, and the present invention is such an action/principle. Although not limited to this range, the following can be considered.
That is, the low-temperature vacuum extraction method is an “extraction method that includes a step of stirring while crushing with a stirring blade, and decompressing while applying heat from outside while crushing and stirring”, and impurities are included in the extraction step. It will not be done. Further, since the extraction is performed at a lower temperature than the conventional extraction method, it is possible to extract components such as natural benzaldehyde which are difficult to be extracted by the conventional extraction method (components weak to heat and volatile components). In addition, a low-temperature vacuum extraction method that does not require the use of an extraction solvent makes it possible to obtain plant-derived high-concentration cell water, and as a result, it is possible to obtain a natural benzaldehyde water containing benzaldehyde and a plurality of active ingredients. ..

Although it is already known that benzaldehyde has an antitumor effect by inhibiting tyrosine kinase activity, the natural benzaldehyde water of the present invention contains other molecules including unidentified molecules in addition to benzaldehyde. Contains as an ingredient. The action/effect of components other than these benzaldehydes, or the synergistic action/effect of these multiple active ingredients including benzaldehyde, enhances the inhibition of tyrosine kinase activity, or by other action, the It is considered that the effect was exhibited.

Hereinafter, the present invention will be described in more detail with reference to Examples and Comparative Examples, but the present invention is not limited to these Examples as long as the gist thereof is not exceeded.

Example 1
<Production of natural benzaldehyde water>
The petals and leaves of the harvested double cherry blossoms were put in the containers shown in FIGS. 1 to 3 for extraction.
The extraction conditions were as follows.
(1) Temperature of Yaezakura (petals or leaves): 30-40°C
(2) Set temperature in container: 30-40°C
(3) Pressure: 93 to 97 kPa lower pressure than 101.3 kPa (1 atm) (4) Rotation speed of stirring blade (movable blade): 4 rpm (rotation/minute)

Of the recovered liquid collected in the recovery tank 41, the liquid in the aqueous layer (aqueous phase) 51 was used as the target extract (natural benzaldehyde water).
8 kg of natural benzaldehyde water was collected from 10 kg of the petals of the double cherry blossoms harvested.

Example 2
<Component analysis of natural benzaldehyde water>
The ratios of the components contained in the natural benzaldehyde water extracted by the low temperature vacuum extraction method were compared by using the gas chromatography method.
The result of natural benzaldehyde water extracted from the petals of double cherry blossoms (hereinafter sometimes abbreviated as “natural benzaldehyde water 1”) is shown in FIG. 4, and the result of natural benzaldehyde water extracted from the leaves of double cherry blossoms (hereinafter referred to as “natural benzaldehyde water 2”) is shown. May be abbreviated as ")" is shown in FIG.
4 and 5, the horizontal axis represents the retention time (minutes) and the vertical axis represents the ionic strength.

From the results of FIG. 4, it was found that the natural benzaldehyde water 1 contained a large amount of benzaldehyde, and also contained a small amount of benzyl alcohol and coumarin.
Further, from the results of FIG. 5, while the main component of the natural benzaldehyde water 2 was benzaldehyde, the content of benzyl alcohol and coumarin was larger than the amount of benzaldehyde compared to the natural benzaldehyde water 1 extracted from the petals.
Further, as can be seen from FIGS. 4 and 5, in addition to benzaldehyde, benzyl alcohol, and coumarin, many other components were detected in both natural benzaldehyde waters 1 and 2. However, it was not possible to identify the detected components.

Example 3
<Measurement of antioxidant and anti-inflammatory effects of natural benzaldehyde water>
The antioxidant and anti-inflammatory effects of natural benzaldehyde waters 1 and 2 were measured using the electron spin resonance method (ESR method).
A sample adjusted to a certain concentration (natural benzaldehyde water), distilled water, a spin trap agent (2,2,6,6-tetramethylpiperidinol; 4-OH TEMP), and a radical generator were added in that order. Mixed.
An appropriate amount of the mixed solution is transferred to an ESR cell, and irradiated with microwaves to superoxide anion radical (O ₂ ^·− ), singlet oxygen ( ¹ O ₂ ), hydroxy radical (HO ^· ) and nitric oxide (NO). The erasing activity (%) was measured.
FIG. 6 shows the superoxide anion radical scavenging activity, FIG. 7 shows the singlet oxygen scavenging activity, FIG. 8 shows the hydroxy radical scavenging activity, and FIG. 9 shows the results of measuring the nitric oxide scavenging activity.

The ordinates in FIGS. 6 to 9 represent the erasing activity (%), and the molecular weight to be measured remaining in the ESR cell when pure water was used as a control was set to 100%.
The “flower” on the horizontal axis indicates the natural aldehyde water 1 extracted from the petals, and the “leaf” indicates the natural aldehyde water 2 extracted from the leaves.
The ratio on the horizontal axis is the ratio of the stock solution when the natural aldehyde water stock solution extracted by the low temperature vacuum extraction method is diluted with distilled water. For example, in FIG. 6, “flower 20%” refers to natural benzaldehyde water 1 diluted 5 times with distilled water.

From the results of FIG. 6, it was found that both the natural benzaldehyde waters 1 and 2 have superoxide anion radical (O ₂ ^·− ) scavenging activity. In particular, it was found that the natural benzaldehyde water 2 extracted from leaves has a higher superoxide anion radical scavenging activity than the natural benzaldehyde water 1 extracted from petals.

From the results of FIG. 7, it was found that the natural benzaldehyde water 1 has a singlet oxygen ( ¹ O ₂ ) scavenging activity, and the natural benzaldehyde water 2 does not have a singlet oxygen scavenging activity. In addition, it was found that the natural benzaldehyde water is superior in the superoxide anion radical scavenging activity to the singlet oxygen scavenging activity.

From the results of FIG. 8, natural benzaldehyde water 1 and 2 were both found to have a hydroxy radical (HO ^·) scavenging activity. Further, it was found that both the natural benzaldehyde waters 1 and 2 obtained by diluting the stock solution of the extract 40 times (“2.5%”) sufficiently exhibited the hydroxy radical scavenging activity.

From the results of FIGS. 6 to 8, it was found that both the natural benzaldehyde waters 1 and 2 have an antioxidant effect.

Further, from the results of FIG. 9, it was found that both natural benzaldehyde waters 1 and 2 have a nitric oxide (NO) scavenging activity, and both natural benzaldehyde waters 1 and 2 have an anti-inflammatory effect. It was

Example 4
<Action of natural benzaldehyde water on saliva>
After diluting the natural benzaldehyde water 1 obtained above with distilled water, a subject (healthy person) is allowed to smell for about 20 minutes using an ordinary commercially available diffuser, and saliva of the subject before and after sniffing is used. Compared. The results are shown in FIGS. 10 and 11.
10 and 11, in the horizontal axis of the graph, "before the experiment" is the saliva of the subject before sniffing the natural benzaldehyde water 1, and "after aroma" is immediately after sniffing the natural benzaldehyde water 1 The saliva of the subject, “after 30 minutes”, shows the saliva of the subject 30 minutes after the sniffing of the natural benzaldehyde water 1 was finished.
Further, “*” in the figure indicates that p<0.05.

10A is the result of measuring the amount of saliva, FIG. 10B is the result of measuring the pH of saliva, and FIG. 10C is the result of measuring the residual amount of superoxide anion radicals (O ₂ ^·− ) in saliva.
From the results of FIGS. 10A to 10C, no change was observed in the saliva amount, the pH, and the residual amount of superoxide anion radicals (O ₂ ⁻ ) by sniffing the natural benzaldehyde water 1.

FIG. 11A shows the result of a BAP test (Biological Anti-oxidant Potental; measurement of antioxidant power). The larger the measured value, the higher the antioxidant power in saliva.
The saliva of the subject 30 minutes after sniffing the natural benzaldehyde water 1 (“30 minutes later”), before sniffing the natural benzaldehyde water 1 (“before experiment”), and immediately after sniffing the smell The measured value was higher than that of saliva (“after aroma”) of the subject.
From the results of FIG. 11A, it was found that smelling natural benzaldehyde water 1 improves the antioxidant ability of human saliva. From the results of FIG. 10C, it was also found that the antioxidant ability was not directly related to the improvement of the scavenging action of superoxide anion radicals.
It is also suggested that natural benzaldehyde water 1 having a composition similar to that of natural benzaldehyde water 1 similarly has an effect of improving the antioxidant ability of human saliva by making it smell.

FIG. 11B shows the results of measuring the amount of cortisol, which is a stress hormone in saliva.
The saliva of the subject 30 minutes after sniffing the natural benzaldehyde water 1 (“30 minutes later”), before sniffing the natural benzaldehyde water 1 (“before experiment”), and immediately after sniffing the smell The amount of cortisol was lower than that of saliva (“after aroma”) of the subjects.
From the results of FIG. 11B, it was found that the amount of cortisol, which is a stress hormone, was suppressed by sniffing the odor of natural benzaldecht water 1.
Similarly, it is suggested that natural benzaldehyde water 1 having a composition similar to that of natural benzaldehyde water 1 has an effect of suppressing the amount of cortisol in saliva by smelling it.

Example 5
<Measurement of antitumor activity of natural benzaldehyde water>
The antitumor activity of the natural benzaldehyde waters 1 and 2 obtained above was verified using the MTT assay.
The MTT assay is a method for measuring the number of living cells based on the fact that the yellow tetrazolium salt, MTT, is taken up by metabolically active living cells and cleaved into purple formazan crystals.
In the test, human alveolar basal epithelial adenocarcinoma cells A549, human squamous cell carcinoma cells A431, and human cervical cancer cells HeLa were used. A549 cells were cultured in 10% (v/v) FBS-containing DMEM medium, and A431 and HeLa cells were cultured in 10% (v/v) FBS-containing MEM medium.

Each cell was seeded on a 96-well culture plate at 3×10 ⁴ cells/100 μL/well, and precultured at 37° C., 5% CO ₂ for 24 hours. After the preculture, the medium was replaced with a test medium, and the culture was performed for 48 hours under the conditions of 37° C. and 5% CO ₂ . Sakura flower cell extract (natural benzaldehyde water 1) and cherry leaf cell extract (natural benzaldehyde water 2), which are evaluation samples, have a final concentration of 0.0 to 5.0% (v/v) in the test medium. Was added respectively.
As a control, a solution prepared by diluting benzaldehyde (Nacalai Tex) with PBS to a concentration of 0.004% to a final concentration of 0.0 to 5.0% (v/v) was used. .. This concentration of benzaldehyde of 0.004% is almost the same as the benzaldehyde content obtained from the results of the component analysis of the cherry blossom cell extract.

After the culture for 48 hours, 10 μL/well of MTT labeling reagent was added, and the mixture was kept warm in an incubator for 4 hours. Then, a solubilization solution (0.01 M HCl, 10% SDS) was added at 100 μL/well and kept warm for 17 hours in an incubator. After keeping the temperature, the absorbance at 570 nm was measured with a reference wavelength of 650 nm using a microplate reader. The cell viability (%) when the evaluation sample was added was determined by setting the number of viable cells when the evaluation sample was not added to 100. The measurement was repeated three times and the average value was calculated.

The results of the human tumor cell growth inhibitory effect of the cherry blossom and leaf cell extract by MTT assay are shown in FIGS. FIG. 12 shows A549 cells, FIG. 13 shows A431 cells, and FIG. 14 shows HeLa cells. The horizontal axis of each graph represents the concentration (%) (v/v) of each sample, and the vertical axis represents the cell survival rate (%). Show.

From the results shown in FIGS. 12 to 14, almost no tumor cell inhibitory effect could be confirmed with the cherry blossom cell extract and the control benzaldehyde, whereas in the cherry leaf cell extract, the tumor cell proliferation inhibitory effect was observed. However, it could be confirmed in any of A549 cells (FIG. 12), A431 cells (FIG. 13) and HeLa cells (FIG. 14).

Natural benzaldehyde water produced using the method for producing natural benzaldehyde water of the present invention has a different component composition from the extract from plants belonging to the genus Rosaceae up to now, natural benzaldehyde as an active ingredient. Including. Further, the natural benzaldehyde water of the present invention is of plant origin, and it is not necessary to use an extraction solvent in the extraction step, so the safety is extremely high.
Therefore, the natural benzaldehyde water is widely used not only in the field of pharmaceuticals but also in the fields of aromatherapy products, fragrances, care products, cosmetics, foods and the like.

1 Container 2 Cooling Tank 6 Stirring Blade 7 Lower Semi-Cylindrical Part 8 Upper Square Part 9 Steam Chamber 10 Discharge Port 14 Exhaust Port 16 Piping 17 Input Port 18 Lid 20 End Wall 21 End Wall 22 End Plate 23 End Plate 24 Blade Body 24a Groove 24b movable blade 25 blade plate 25a groove 25b movable blade 26 fixed blade 30 inclined surface 31 plant belonging to Rosaceae genus Prunus (Prunus genus plant)
32 vacuum gauge 33 thermometer 34 thermometer 41 recovery tank 45 valve 46 pressure reducing device 49 valve 50 oil layer 51 water layer R rotational direction

Claims (9)

A method for producing an antitumor agent containing natural benzaldehyde water,
The natural benzaldehyde water was stirred while crushing the cherry leaves with a stirring blade, and heat was applied from the outside under the crushing/stirring to maintain the temperature of the cherry leaves at 50° C. or lower while maintaining a temperature of 101.3 kPa ( The method for producing an antitumor agent is characterized in that the antitumor agent is produced by an extraction method having a step of decompressing while maintaining a pressure lower than 80 kPa per 1 atmosphere) . A method for producing an antioxidant containing natural benzaldehyde water,
The natural benzaldehyde water was stirred while crushing the cherry leaves with a stirring blade, and heat was applied from the outside under the crushing/stirring to maintain the temperature of the cherry leaves at 50° C. or lower while maintaining a temperature of 101.3 kPa ( The method for producing an antioxidant is characterized in that it is produced by an extraction method having a step of decompressing while maintaining a pressure lower than 80 kPa per 1 atmosphere) . The above antioxidant is a superoxide anion radical (O _Two ^・− ) The method for producing an antioxidant according to claim 2, which is an erasing activator. The method for producing an antioxidant according to claim 2, wherein the antioxidant is a hydroxyl radical (HO.) scavenging activator. A method for producing an anti-inflammatory agent containing natural benzaldehyde water,
The natural benzaldehyde water was stirred while crushing the cherry leaves with a stirring blade, and heat was applied from the outside under the crushing/stirring to maintain the temperature of the cherry leaves at 50° C. or lower while maintaining a temperature of 101.3 kPa ( The method for producing an anti-inflammatory agent is characterized in that the anti-inflammatory agent is produced by an extraction method having a step of decompressing while maintaining a pressure lower than 80 kPa per 1 atmosphere) . The method for producing an anti-inflammatory agent according to claim 5, wherein the anti-inflammatory agent is a nitric oxide (NO) scavenging activator. An antitumor agent, which is produced using the method for producing an antitumor agent according to claim 1. An antioxidant, which is produced using the method for producing an antioxidant according to any one of claims 2 to 4. An anti-inflammatory agent produced by using the method for producing an anti-inflammatory agent according to claim 5 or 6.

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