Composition for promoting collagen production, method for producing the composition for promoting collagen production, and external preparation for skin

Abstract

To provide a composition for promoting collagen production, which uses a substance obtained from olives and is capable of exhibiting the function of promoting collagen production in human cells, a method for producing the composition for promoting collagen production, and a skin external preparation comprising the composition for promoting collagen production.SOLUTION: Provided is a composition for promoting collagen production. The composition is provided for improving the collagen production function of human cells and comprises a distillate obtained from olive leaves. The composition is capable of promoting the collagen-producing function of skin cells, allowing the skin cells to exhibit an anti-aging function.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a composition for promoting collagen production, a method for producing the composition for promoting collagen production, and an external preparation for skin. More specifically, it relates to a composition for promoting collagen production obtained from olive leaves, a method for producing the composition for promoting collagen production, and an external preparation for skin.

Background Art

[0002] In recent years, due to the increasing demand for safety and peace of mind in cosmetics and the trend towards an environmentally conscious lifestyle, the demand for botanical cosmetics containing plant-derived ingredients has been increasing. In particular, in recent years, with the advent of an aging society, cosmetics with anti-aging functions are in demand, and as one of their functions, the development of a collagen production promoting function that promotes the production of skin collagen is required.

[0003] Here, there is the olive, a plant that has been used in various functional foods and the like since ancient times. It is known that olives contain many polyphenols, and the functions of olive polyphenols have been clarified. For example, Patent Document 1 discloses a cosmetic containing an olive fruit extract, and it is described that by containing oleuropein and hydroxytyrosol, it can exhibit an action of suppressing collagen degradation. Patent Document 2 describes that acteoside, a kind of polyphenols contained in olive fruits, has α-glucosidase inhibitory activity. Patent Document 3 describes that luteolin, a kind of polyphenols contained in olive leaf extracts, has amylase inhibitory activity.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Patent Document 2

Patent Document 3

Disclosure of the Invention

Problems to be Solved by the Invention

[0005] However, in the conventional technology, it has not been clarified whether the substance obtained from olives has a function of promoting the production of collagen in human cells.

[0006] In view of the above circumstances, the present invention provides a composition for promoting collagen production that can exert a function of promoting the production of collagen in human cells using a substance obtained from olives, a method for producing the composition for promoting collagen production, and a topical skin preparation containing the composition for promoting collagen production.

[0007] As a result of intensive studies to solve the above problems, the present inventors have found that a substance obtained from olives has an effect of promoting the production of collagen in human cells, and have thus completed the present invention.

Means for Solving the Problems

[0008] The composition for promoting collagen production of the present invention is a composition for improving the collagen production function of human cells, and is characterized by containing a distillate obtained from olive leaves. The method for producing the composition for promoting collagen production of the present invention is a method for producing a composition for improving the collagen production function of human cells, and is characterized by including a step of obtaining a distillate while subjecting olive leaves to vacuum distillation. The topical skin preparation of the present invention is characterized by containing the composition for promoting collagen production of the present invention.

[0009] According to the present invention, when applied to human skin, the collagen production function of skin cells can be promoted, so that an anti-aging function can be exerted on skin cells.

Brief Description of the Drawings

[0010]

Figure 1

Figure 2

Figure 3

Figure 4

Embodiments for Carrying Out the Invention

[0011] Embodiments of the present invention will be described based on the drawings. The composition for promoting collagen production of this embodiment contains the distillate of olive leaves. If this composition for promoting collagen production is applied to human skin, it can be expected to suppress the aging of human skin cells.

[0012] In the specification, an olive is a plant belonging to the genus Olea of the family Oleaceae, and its type is not particularly limited. The target part of the olive is the leaf, and these can be used alone or in combination. Hereinafter, unless otherwise specified, when referring to an olive, it means the above-mentioned target part.

[0013] The cultivation state of the target olive is not particularly limited. For example, if leaves attached to olive branches that are scheduled to be pruned and discarded during the fruit harvesting process of olives are used, the advantage of effectively utilizing unused biomass resources can be obtained.

[0014] (The composition for promoting collagen production of this embodiment) The distillate contained in the composition for promoting collagen production of the present embodiment (hereinafter referred to as the composition for promoting collagen production) is obtained by heating and distilling olive leaves. Specifically, this distillate is a liquid obtained by heating olive leaves and cooling the generated gas or the like. This distillate contains, for example, an oily substance (so-called essential oil) with a relatively small molecular weight and a water-soluble substance.

[0015] If the composition for promoting collagen production is supplied to human cells, it can promote the function of producing collagen (collagen production function) possessed by the cells. In other words, the composition for promoting collagen production contains the above distillate as an active ingredient for promoting the collagen production function possessed by human cells. Note that this active ingredient means a component that can promote the collagen production function. Further, this component includes one or more substances. Therefore, if a composition for promoting collagen production containing the above distillate is applied to human skin by coating or the like, it is possible to suppress skin aging. That is, the composition for promoting collagen production has an anti-aging function against the aging phenomenon of human skin cells. In other words, the composition for promoting collagen production can be used in cosmetics such as skin care products.

[0016] The form of the composition for promoting collagen production is not particularly limited. For example, forms such as liquid, gel, cream, etc. can be mentioned. Also, it may be in a form filled in a sprayer or the like and applied as fine particles.

[0017] Here, vitamin C (ascorbic acid) is known as a substance that promotes the collagen production function of human cells (human cells). On the other hand, it has been reported that polyphenols obtained from olives (including parts other than leaves) have functions such as suppressing collagen-degrading enzymes and having α-glucosidase inhibitory activity and amylase inhibitory activity. However, it has not been reported that substances obtained from olives (including parts other than leaves) have the function of promoting the collagen production function of human cells. That is, the inventors have found for the first time a component that acts on human cells and promotes the collagen production function from olives (including parts other than leaves). Moreover, it has also been found for the first time that such an active ingredient has a smaller molecular weight compared to polyphenols.

[0018] The distillate of the composition for promoting collagen production is obtained by subjecting olive leaves to heat distillation treatment as described above, and mainly consists of a group of volatile substances contained in olive leaves.

[0019] Note that the composition for promoting collagen production may contain other components as long as the effect of promoting the collagen production function of human cells is not impaired. For example, preservatives, stabilizers, pH adjusters, etc. may be appropriately blended. Also, the composition for promoting collagen production may be appropriately used as an additive according to the application. For example, if the composition for promoting collagen production is added to a moisturizer, etc., in addition to the effect of moisturizing the skin, the effect of improving the firmness of the skin can be obtained. Note that examples of the moisturizer include those mainly composed of polyhydric alcohols such as glycerin and butylene glycol.

[0020] (External preparation for skin of this embodiment) The external preparation for skin of this embodiment contains a composition for promoting collagen production containing the distillate obtained from the above-mentioned olive leaves. Therefore, by applying the external preparation for skin of this embodiment, etc., to impart the composition for promoting collagen production contained in the external preparation for skin of this embodiment to human skin, the collagen production function of skin cells can be promoted, so that the skin can be maintained in a youthful state. That is, if the external preparation for skin of this embodiment is applied to the skin, etc., the anti-aging function can be exerted or improved on skin cells. Therefore, the external preparation for skin of this embodiment can be used as cosmetics such as skin care products.

[0021] In addition, the skin external preparation of the present embodiment may contain other components (for example, moisturizing components, etc.) in addition to the composition for promoting collagen production, and can be appropriately adjusted according to the intended use.

[0022] (Method for producing the composition for promoting collagen production of the present embodiment) Next, the method for producing the composition for promoting collagen production of the present embodiment (hereinafter referred to as the method for producing the composition for promoting collagen production) will be described.

[0023] The method for producing the composition for promoting collagen production is a method for producing a composition for improving the collagen production function of human cells (human cells), and includes a step of obtaining a distillate while subjecting olive leaves to vacuum distillation (hereinafter referred to as the distillate recovery step).

[0024] This distillate recovery step is not particularly limited as long as a group of substances that are easily volatilized from olive leaves can be recovered as the main component. Specifically, it is preferable to adopt a method in which non-volatile components such as solid components and polyphenols of olive leaves and hardly non-volatile components such as lignin are separated as much as possible. For example, in the distillate recovery step, for example, a generally used distillation apparatus, a microwave distillation apparatus, or the like can be adopted. Such an apparatus can accommodate olive leaves in a housing portion, perform a predetermined operation, separate solid components and a distillate from the olive leaves, and recover the distillate. The obtained distillate corresponds to the distillate contained in the composition for promoting collagen production. This distillate may be used as it is as the composition for promoting collagen production, or may be diluted and adjusted, or prepared by blending other substances.

[0025] The olive leaves supplied to the apparatus in the above distillate recovery step may be in the state as separated from the branches or in a smaller state. Known methods can be used to make the olive leaves smaller. For example, by performing treatments such as cutting or grinding, they can be used as ground materials such as chips or powders, which makes it possible to improve the handleability.

[0026] Note that the olive leaves to be used may be supplied to the distillate recovery step in a state where they have been supplied to a drying step and dried before being supplied to the above apparatus. In this case, it becomes possible to improve the operability when supplying to the apparatus. However, it is preferable to adjust the drying conditions in this drying step so as not to significantly impair the recovery rate of the above distillate. For example, freeze-drying or low-temperature drying can also be used for drying.

[0027] Also, before the distillate recovery step, a step of obtaining a squeezed juice using a squeezer for the olive leaves may be provided. Then, the obtained squeezed olive leaves and / or the squeezed juice are supplied to the distillate recovery step. In this case, an improvement in the recovery rate of the distillate can be expected. Furthermore, a step of irradiating the olive leaves with microwaves may be provided before the step of obtaining the squeezed juice. In this case, since the cell walls constituting the olive leaves are in a state where they are easily broken by the microwaves, the advantage is obtained that components that are easily volatilized can be easily obtained.

[0028] The treatment using the distillation apparatus in the distillate recovery step can be carried out by atmospheric distillation, vacuum distillation, steam distillation, etc. And the heating method when using the distillation apparatus is not particularly limited, and for example, in addition to heaters, hot water, and steam, a method of irradiating microwaves can be adopted. The method of vacuum distillation while irradiating microwaves corresponds to the method of "vacuum distillation while irradiating microwaves" in the method of the claims.

[0029] When the method of irradiating microwaves is adopted, the advantage is obtained that components that are easily volatilized contained in the olive leaves can be easily obtained. Specifically, when the olive leaves are irradiated with microwaves, the moisture contained in the olive leaves is heated and released as water vapor. At this time, volatile or easily volatile components (such as essential oils and water-soluble compounds) contained in the olive leaves are obtained as distillates. Therefore, it is possible to efficiently recover volatile components and the like present inside the cell wall, which are difficult to extract by general distillation methods. That is, by adopting the method of irradiating microwaves, volatile components and the like contained in the olive leaves can be efficiently obtained.

[0030] As the distillation apparatus for irradiating microwaves, a known one can be adopted as described above.

[0031] For example, a microwave vacuum distillation apparatus 1 as shown in FIG. 1 can be used. This microwave vacuum distillation apparatus 1 includes a distillation tank 2 corresponding to a housing part for housing a distillation object 11 (olive leaves) as a raw material, a microwave heating apparatus 3 for irradiating microwaves, an air flow inlet pipe 4 for supplying an inert gas such as air or nitrogen gas to the distillation tank 2, a distillate outflow pipe 5 for guiding the distilled distillate, a cooling apparatus 6 for cooling the distillate outflow pipe 5, a heating control apparatus 7 for controlling the temperature of the distillation tank 2, and a pressure control apparatus 10 for controlling the pressure inside the distillation tank 2. The pressure control apparatus 10 includes a vacuum pump 8 and a pressure control valve 9. And the microwave vacuum distillation apparatus 1 is configured such that the pressure and the like inside the distillation tank 2 can be adjusted via the heating control apparatus 7 and the pressure control apparatus 10, respectively.

[0032] Therefore, when the olive leaves are housed in the distillation tank 2 of the microwave vacuum distillation apparatus 1 and the microwave vacuum distillation apparatus 1 is operated, the gaseous gas containing volatile components and the like contained in the olive leaves is changed into a liquid by the cooling apparatus 6 via the distillate outflow pipe 5 and obtained as a distillate 12. This distillate 12 corresponds to the distillate of the composition for promoting collagen production described above.

[0033] In addition, as the distillation conditions when using a distillation apparatus, known conditions can be adopted. The conditions vary depending on the internal volume, its water content, the intensity of microwaves, etc. For example, the pressure inside the accommodation part of the apparatus can be adjusted so that the pressure is under a reduced pressure of 10 to 90 kPa and the treatment time is 0.2 to 8 hours.

[0034] In addition, the distillate may be used after being separated into an essential oil part and a water-soluble part, or may be used in a mixed state. That is, the one in the state where the essential oil part and the water-soluble part are mixed may be adopted as the distillate of the composition for promoting collagen production, or the separated ones may be adopted separately as the distillates of the composition for promoting collagen production. The method for separating the essential oil part and the water-soluble part is not particularly limited. For example, the mixed one can be separated into an oily fraction and a water-soluble fraction by performing treatments such as allowing it to stand, subjecting it to redistillation treatment, and liquid-liquid separation.

Examples

[0035] Next, the present invention will be described in more detail by way of examples. Note that these examples show an example of this embodiment, and the present invention is not limited in any way by the following examples.

[0036] The crushed product obtained by crushing olive leaves with a crusher was put into the distillation tank of the microwave vacuum distillation apparatus shown in Fig. 1, the inside of the distillation tank was set to the following conditions, and residues and distillates were obtained. Pressure inside the distillation tank: reduced pressure condition of 0.25 kPa (minus 75 kPa) Distillation time: 30 minutes

[0037] The evaluation of the collagen production promoting function of the obtained distillate was carried out as follows.

[0038] Diluent: EMEM medium of serum-free medium Cells used: KMST-6 (human fetal-derived fibroblasts, Riken Cell Bank RCB1955) Containers used: Microplate 12well Control (Ct): EMEM medium without serum

[0039] <Method for evaluating the function of promoting collagen production> (1: Cell culture) KMST-6 subcultured in a tissue culture flask (25 cm 2 ) was cultured in EMEM medium (manufactured by Fujifilm Wako Pure Chemical Corporation) containing 10% FBS (fetal bovine serum, manufactured by Gibco Life technology). After culturing for several days, the confluent cells were seeded into a 12-well microplate at 1 ml per well. After seeding, pre-culture was performed for 3 days.

[0040] (2: Addition of distillate to fibroblasts) The distillate was diluted to a predetermined magnification (2-fold, 4-fold) using EMEM medium without serum to prepare a 2-fold diluent of the distillate and a 4-fold diluent of the distillate. Ascorbic acid was added to each diluent so that the concentration was 75 μM (μmol / l). 1 ml of each prepared diluent was added to each well after pre-culture. As Ct, EMEM medium without serum was used. After addition, culture was performed for 3 days.

[0041] (3: Recovery of collagen) After culturing, the culture supernatant was removed. Trypsin solution was added to the plate from which the supernatant had been removed to partially decompose the collagen fibers, and the cells remaining on the plate were detached and recovered from the inner wall surface of the plate. In this recovered solution, the collagen fibers partially decomposed by trypsin treatment were recovered in a solubilized state. This recovered solution was transferred to a centrifuge tube and centrifuged (10,000 rpm, 4 °C, 5 min), and then the supernatant was recovered. Since this supernatant contained collagen fibers, this supernatant was used as a sample for measuring the amount of collagen fibers. On the other hand, 1 ml of EMEM medium was added to a centrifuge tube from which the supernatant had been removed to prepare a cell suspension. To 100 μl of this cell suspension, 100 μl of 0.5% trypan blue staining solution (manufactured by Nacalai) was added to perform trypan blue staining. A counting chamber was used to measure the number of viable cells.

[0042] (4: Quantification of collagen) A staining solution was prepared by mixing 1 mg / ml aqueous Direct Red 80 solution (manufactured by Sigma-Aldrich) and saturated aqueous picric acid solution (manufactured by Nacalai Tesque) at a ratio of 1:1. 100 μl of a sample for measuring the amount of collagen fibers was dispensed into a 1.5-ml microtube, and 1.0 ml of the staining solution was added. This microtube was gently inverted and mixed for 30 minutes. After mixing, centrifugation (12,000 rpm, 4°C, 10 min) was performed to remove the supernatant. 1.0 ml of 0.5 M NaOH (0.5 mol / L) aqueous solution was added to the microtube from which the supernatant had been removed, and it was stirred using a vortex mixer. The solution after the stirring was completed was dispensed into a 96-well microplate in 200-μl aliquots, and the absorbance was measured at 550 nm. The calibration curve was prepared using porcine skin-derived acid-soluble type I collagen dissolved in 0.5 M acetic acid solution to a concentration of 0.05 - 1.0 μg / μl. The quantification of collagen was calculated using the following formula. First, the absorbance of the sample for measuring the amount of collagen fibers was applied to the calibration curve to calculate the amount of collagen per volume. Then, the calculated value was divided by the number of viable cells measured in (3: Collagen recovery) above to calculate the amount of collagen per cell. Amount of collagen per cell (μg / cell) = (Amount of collagen per volume (μg / ml)) / (Number of viable cells (ml / cells))

[0043] The evaluation results of the collagen production promoting function are shown in Figure 2. It was confirmed that the extract obtained from olive leaves can increase the amount of collagen fibers in human cells by about four times compared to Ct (control).

[0044] <Component analysis in the distillate> A certain amount of the distillate was aliquoted into vials, and the components in the distillate were analyzed. The apparatus conditions used are as follows. Apparatus: Headspace gas chromatograph mass spectrometer (HS-GC / MS; manufactured by JEOL Ltd., 6890N / JMS-Q1000GC) Column: HP-5MS (25 m × column inner diameter 250 μm, film thickness 0.25 μm) Column oven: 40 °C (2 min) → 5 °C / min → 90 °C (0 min) → 10 °C / min → 200 °C (5 min) Inlet: 200 °C, Interface: 230 °C Ion source: 230 °C Sample injection: Headspace method HS oven: 80 °C Desorption: 2 min Dry purge: 8 min Sample amount: 10 ml (NaCl: 30%) Cyclohexanol was added as an internal standard substance.

[0045] The analysis results are shown in Table 1. An example of the chromatogram is shown in Figure 3. The concentration of each detected substance in the distillate was expressed as a cyclohexanol equivalent value (mg / l) based on each peak in the chromatogram and the peak of cyclohexanol added as an internal standard substance. From Table 1, it can be confirmed that the distillate contains a wide variety of volatile components.

[0046]

Table 1

[0047] <Analysis of polyphenols in the distillate> The polyphenols in the distillate were evaluated in terms of the total polyphenol content. The total phenolic content (mg-GAE / 100 g of distillate) was evaluated as the equivalent amount of gallic acid (mg-gallic acid) per 100 g of distillate by the Folin-Ciocalteu method.

[0048] The measurement conditions were as follows. Detector: Microplate reader (manufactured by Corona Electric Co., Ltd., model number: SH-1200 Lab) Measurement wavelength: 750 nm Standard reagent for calibration curve: Gallic acid (manufactured by Wako Pure Chemical Industries, Ltd.) Folin-Ciocalteu reagent: Manufactured by Wako Pure Chemical Industries, Ltd.

[0049] As a comparative example, the total polyphenol content was measured in the same manner as in the examples, except that an extract of the residue obtained when treated with a microwave vacuum distillation apparatus (residue extract) and an extract of olive leaves that had been heat-dried (heat-dried extract) were used. The residue extract was prepared by pulverizing the residue with a mill (manufactured by Osaka Chemical Co., Ltd., Wonder Blender WB-1), adding 5 g of the obtained powder to 100 ml of a 70% ethanol aqueous solution, shaking for 60 minutes, allowing to stand, and then collecting the supernatant. The heat-dried extract was prepared by heat-drying olive leaves at 60 °C for 1 day using an oven and treating them in the same manner as the residue extract.

[0050] Figure 4 shows the analysis results of the total polyphenols. As shown in Figure 4, it was confirmed that the distillate contained almost no polyphenols. On the other hand, it was confirmed that the residue extract and the heat-dried extract of the comparative example contained 600 to 1000 times more polyphenols than the distillate.

[0051] From the results of the above experiments, it was confirmed for the first time that the distillate of the composition for promoting collagen production of the present invention can promote the collagen production function in human cells, despite containing almost no polyphenols. This is presumably an effect exerted by the interaction of various components other than the polyphenols of olives contained in the distillate. Therefore, the collagen production promoting function of the composition for promoting collagen production of the present invention can be applied to cosmetics such as skin care products. In addition, the present invention is a newly discovered technology, and it is possible to propose a new method of using the distillate obtained by the vacuum microwave steam distillation method, and it is possible to use olive leaves more effectively.

Industrial Applicability

[0052] The composition for promoting collagen production and the external preparation for skin of the present invention are suitable for preventing skin aging. In addition, the method for producing the composition for promoting collagen production of the present invention is suitable for producing the composition for promoting collagen production of the present invention.

Explanation of Signs

[0053] 1 Microwave vacuum distillation apparatus 2 Distillation tank 3 Microwave heating device 4 Airflow inlet pipe 5 Distillate outflow pipe 6 Cooling device 7 Heating control device 8 Vacuum pump 9 Pressure control valve 10 Pressure control device 11 Object to be distilled 12 Distillate

Claims

1. A composition for improving the collagen-producing function of human cells, containing a distillate obtained from olive leaves by a microwave vacuum distillation method, wherein the distillate does not contain polyphenols with a molecular weight greater than 210.31 and is a composition for promoting collagen production, characterized by this.

2. A method for producing a composition for improving the collagen-producing function of human cells according to Claim 1, comprising a step of obtaining a distillate while subjecting olive leaves to vacuum distillation using a microwave vacuum distillation method and being a method for producing a composition for promoting collagen production, characterized by this.

3. Containing the composition for promoting collagen production according to Claim 1 and being an external preparation for skin, characterized by this.

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