JP2017178850A - Skin external preparation - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a skin external preparation.SOLUTION: The skin external preparation comprises 0.05 to 5 mass% of one or more extract selected from an extract of a female flower head of Humulus lupulus and an extract of a leaf of Origanum majorana (Majorana hortensis) as an active ingredient for angiogenesis inhibitory action.SELECTED DRAWING: None

Description

  The present invention relates to an external preparation for skin.

  It is well known that human skin is roughly divided into three layers: epidermis, dermis, and subcutaneous tissue. Living cells also exist in the epidermis, and when the epidermal keratinocytes in the basal layer divide and mature, they differentiate through the spiny layer and granule layer into the dead layer of the dead cell. At this time, capillaries that reach the dermis carry and supply substances necessary for cell division of epidermal keratinocytes.

It is known that blood vessels are induced during organ formation, wound healing, inflammation repair, and the like, and this is an in vivo phenomenon called angiogenesis.
However, it is already known that when this angiogenesis is performed excessively, deterioration of the skin condition, particularly wrinkle formation is promoted.
Angiogenesis is enhanced by the action of vascular endothelial growth factor (VEGF), and thrombospondin 1 (TSP-1) has been shown to suppress the action, and the expression of VEGF is increased during UV irradiation, and TSP-1 Has been reported to decrease in expression (Non-patent Document 1). That is, the balance between VEGF and TSP-1 is lost, and angiogenesis is promoted by the relative predominance of VEGF.
Since the neutrophils that produce elastase can easily infiltrate the skin tissue from the neovascularization at the site irradiated with ultraviolet rays, the elastin fibers are destroyed. As a result, changes in skin structure, particularly wrinkle formation, are promoted.

Patent Document 1 describes a composition for inhibiting angiogenesis, which contains D-methionine and derivatives thereof (claims).
Patent Document 2 describes an angiogenesis inhibitor containing an extract of Ushiratake, Numerisugitake, and Kuritatake (claims). In the examples, a lotion containing 5.0% by weight of Kuritake extract is described.
Patent Document 3 describes that a fermented barley extract has an anti-angiogenic action, and describes a functional food material containing the extract.

JP 2013-177356 A JP 2007-238454 A JP 2007-84504 A

Journal of Investigative Dermatology 118,800-805, 2002

  An object of the present invention is to provide an external preparation for skin.

  In the present invention, one or more extracts selected from a hop (HUMULUS LUPULUS) female flower spike extract and a marjoram (ORIJANUM MAJORANA = MAJORANA HORTENSIS) leaf extract are used as an effective component for inhibiting angiogenesis. An external preparation for skin containing 05 to 5% by mass is provided.

  The external preparation for skin of the present invention can suppress angiogenesis promoted by ultraviolet irradiation, improve skin elasticity, and give skin firmness.

The result of the human umbilical vein endothelial cell (HUVEC) proliferation rate of Experiment 1 (hop extract) is shown. The result of the cell growth rate using the human umbilical vein endothelial cell (HUVEC) of the test example 1 (marjoram extract) is shown. The result of the human umbilical vein endothelial cell (HUVEC) proliferation rate of Test Example 1 (mixture of hop extract and marjoram extract) is shown.

<External preparation for skin>
The external preparation for skin of the present invention contains one or more extracts selected from an extract of hop female spikelets and an extract of marjoram leaves as an active ingredient for inhibiting angiogenesis.

  A hop female flower spike refers to a female flower spike of a mulberry family hop (HUMULUS LUPULUS). The plant has been used for medicinal purposes since ancient times, and is sometimes used as a bitter taste stomach medicine. Hop related plants can also be used.

  The marjoram leaf used in the present invention refers to a leaf of the family Lamiaceae (ORIGANUM MAJORANA = MAJORANA HORTENSIS). Essential oils and extracts of marjoram are still used for purposes such as sedation and anti-anxiety. Marjoram related plants can also be used.

The extract used in the present invention can be obtained, for example, by drying and chopping hops of female hops or leaves of marjoram, or powdering, adding an extraction solvent, and performing cold immersion extraction or heat extraction.
A well-known method can be adopted as the extraction method by the cold extraction method. For example, the extract used in the present invention can be obtained by adding 1 to 1.5 L of the extraction solvent to 100 g of hop female flower spikes and marjoram leaves and stirring at room temperature.
A well-known method can also be adopted as the extraction method by the heat extraction method. For example, the extract used in the present invention can be obtained by adding 0.5 to 1 L of the extraction solvent to 100 g of hops of female hops and marjoram leaves and heating to reflux.

  As the extraction solvent, a mixed solvent of water and an organic solvent selected from ethanol, 1,3-butanediol, isopropanol and the like can be used.

  The extract in the present invention includes any of an extract, a diluted or concentrated solution of the extract, and a dried product (for example, powder) obtained by drying the extract. Those dissolved in a solvent such as water are also included.

The content of the extract used in the present invention is 0.05 to 5% by mass, preferably 0.06 to 3% by mass, and more preferably 0.1 to 2% by mass from the viewpoint of effective concentration and stability. .
The extract used in the present invention has an excellent angiogenesis-inhibiting action, and a high effect can be obtained in a small amount.

  When the extract used in the present invention is a product obtained by extracting hop female spikes (hop extract), the content of the extract is preferably 0.05 to 3% by mass, more preferably 0. It is 1-2 mass%, More preferably, it is 0.25-2.0 mass%.

  When the extract used in the present invention is obtained by extracting a marjoram leaf (marjoram extract), the content of the extract is preferably 0.3 to 1.75% by mass, more preferably It is 0.5-1.5 mass%, More preferably, it is 0.75-1 mass%.

As the extract used in the present invention, a hop extract and a marjoram extract can be used alone or in combination.
When using a combination of hop extract and marjoram extract,
The content of the hop extract in the external preparation for skin is preferably 0.05 to 3% by mass, more preferably 0.06 to 2% by mass,
The content of the marjoram extract in the external preparation for skin is preferably 0.1 to 1.5% by mass, more preferably 0.12 to 1% by mass.
The ratio of the hop extract and marjoram extract in the total amount is hop extract / marjoram extract (mass ratio), preferably 1/3 to 3/1, more preferably 1/2 to 2/1. / 2 to 1/1 is more preferable.

When the hop extract and marjoram extract are liquids (including concentrates), the external preparation for skin of the present invention can be used as it is as an external preparation for skin, as well as various components blended in the external preparation for skin and Can be combined.
When the hop extract and marjoram extract are dried products (powder, etc.), the external preparation for skin of the present invention is preferably combined with various components blended in the external preparation for skin.

As various ingredients that are generally blended into skin external preparations,
Hydrocarbons, squalane, liquid paraffin, petrolatum, solid paraffin, microcrystalline wax, ceresin, alkyd, acrylic, sulfonamide resin, nitrocellulose;
Mineral oil, olive oil, almond oil, cacao butter, macadamia nut oil, avocado oil, hydrogenated palm oil, castor oil, sunflower oil, evening primrose oil, synthetic triglyceride, wax, sunflower seed wax, beeswax, carnauba wax, candelilla wax, lanolin, acetic acid Lanolin, higher fatty acid, higher alcohol, synthetic ester, linear / cyclic silicone oil, dimethylpolysiloxane, glyceryl monostearate, decamethylcyclopentanesiloxane, polyoxyethylene-modified dimethylpolysiloxane, octamethylcyclotetrasiloxane, polyphenylmethyl Siloxane, rosin pentaerythritol ester, neopentyl glycol diisooctanoate, glycerin triisooctanoate, isopropyl myristate, propylene monolaurate Oil, such as glycol, tri (caprylic acid / capric acid) glyceryl;
Purified water, ethanol, isopropanol, polyhydric alcohol, water-soluble polymer, bentonite, polyoxyethylene sorbitan monostearate, sorbitan sesquiisostearate, ethyl acetate, butyl acetate, butanol; glycerin, propylene glycol, sorbit, polyethylene glycol, di- Moisturizing agents such as propylene glycol, 1,3-butylene glycol, diglycerin, mannitol, POE methyl glycoside, biopolymer, sucrose;
Emollient ingredients such as cetyl ethylhexanoate, diisostearyl malate, dipentaerythrityl tripolyhydroxystearate; quince seed, pectin, cellulose derivatives, xanthan gum, sodium alginate, soagina, carboxyvinyl polymer;
Octyldodecyl myristate, neopentyl glycol diisooctanoate, sorbitan monooleate, glycerin triisooctanoate, octylmethoxycinnamate, POE sorbitan monooleate, isocetyl, isostearic acid, stearic acid, isopropyl palmitate, acetyltributyl citrate, Trimethylpentanediyl dibenzoate, stearylconium bentonite;
Thickeners, preservatives, emulsifiers, stabilizers;
Anionic surfactants, cationic surfactants, nonionic surfactants, amphoteric surfactants, protein surfactants;
Medicinal ingredients, vitamin C dipalmitate, vitamins, amino acids, whitening agents, bactericides;
Talc, kaolin, mica, sericite, alumina, silica, glass flake, calcium carbonate, magnesium carbonate, silicic acid anhydride, barium sulfate, cosmetic tar pigment, organic pigment, β-carotene, calsamine, carmine, chlorophyll, bengara, yellow oxidation Pigments such as iron, black iron oxide, ultramarine, bitumen, chromium oxide, carbon black, white pigment, pearl pigment,
Nylon powder, polyethylene powder, polymethyl methacrylate, wool powder, cellulose powder, silk powder, silicone powder, starch powder, aluminum powder, spherical nylon, spherical polystyrene, mica titanium, titanium dioxide, titanium oxide, iron oxide, other powders body;
pH adjuster, potassium hydroxide, sodium hydroxide, triethanolamine, magnesium stearate, zinc stearate, metal ion sealant, citric acid, sodium citrate, lactic acid, sodium lactate, UV absorber, antioxidant, Examples include fragrances, preservatives, chelating agents, antioxidants, dispersants, browning inhibitors, buffers, precipitation preventing components, gelling agents, organically modified clay minerals, usability modifiers, and the like.

The external preparation for skin of the present invention can take various forms such as lotions, emulsions, creams, ointments, adhesives and the like.
The external preparation for skin of the present invention can be used as a quasi-drug for cosmetics and skin diseases.

  EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples. However, the scope of the present invention is not limited to these examples. “%” Means mass% unless otherwise specified.

Preparation Example 1 (Hop extract)
After washing with water and pulverizing 500 g of hop female panicles, about 5 L of extraction solvent (water / 1,3-butanediol = 1: 1 (mass ratio)) was heated and extracted (refluxed at 80 ° C.). The obtained extract was filtered and separated into a reddish brown extract and straw. The solvent of the obtained extract was distilled off to obtain a hop extract as a paste-like concentrate.

Preparation Example 2 (Marjoram extract)
500 g of marjoram leaves washed with water and pulverized are obtained by heating (80 ° C. reflux heating) using about 5 L of extraction solvent (water / 1,3-butanediol = 1: 1 (mass ratio)). The resulting extract was filtered and separated into a tan extract and straw. The solvent of the obtained extract was distilled off to obtain a marjoram extract as a paste-like concentrate.

Preparation Example 3 (mixture of hop extract and marjoram extract)
The extracts of Preparation Examples 1 and 2 were mixed at an arbitrary ratio to obtain a brown mixed extract. The solvent of the obtained mixed extract was distilled off to obtain a mixture of a hop extract and a marjoram extract as a paste-like concentrate.

Test Example 1 (Evaluation of angiogenesis inhibitory effect)
Human umbilical vein endothelial cells (HUVEC) were used using the hop extract of Preparation Example 1 alone, the marjoram extract of Preparation Example 2 alone, and the mixture of the hop extract and Marjoram extract of Preparation Example 3 as samples. Each angiogenesis inhibitory effect was evaluated by the cell proliferation rate.
First, the following (i) intravascular cell basal medium 1, (ii) intravascular cell basal medium 2 and (iii) coloring solution were prepared.

(I) Preparation of vascular cell basal medium 1 To HuMedia-EB2 (manufactured by Kurashiki Boseki Co., Ltd.), FBS (final concentration 2% by volume), 10 ng / mLhEFG, 1.34 μg / mLHC, 50 μg / mLGM, 50 ng / mLATB, 5 ng / MLhFGF-B, 10 μg / mLHP was added.

(Ii) Preparation of Intravascular Cell Basal Medium 2 To vascular endothelial cell basal medium 1 of (i) above, 5 ng / mL VEGF was added and prepared.

(Iii) Preparation of coloring solution [3- (4,5-Dimethylthial-2-yl) -2,5-DiphenyltetrazaliumBromide] (MTT) phosphate buffered saline (PBS (-) to 5 mg / mL ) And dissolved.

Using the vascular cell basal medium 1, the HUVEC was adjusted to 1.5 × 10 ⁶ cells / mL, seeded in 100 μL each in a 96-well microplate, 5% carbon dioxide gas, saturated water vapor, 37 Incubated at 0 ° C.

  After 24 hours, the culture solution was removed by suction, and each sample (50 mass% 1,3-butanediol aqueous solution of each extract) was added to the vascular cell basal medium in the amount shown in FIGS. 2 and cultured at 37 ° C. under 5% carbon dioxide gas and saturated water vapor.

After 24 hours, the culture solution was removed by aspiration, 100 μL of a 10-fold diluted color developing solution in vascular cell basal medium 1 was added and cultured at 37 ° C. in 5% carbon dioxide gas and saturated water vapor.
After 2 hours, the diluted color developing solution was removed, the dye was extracted with 150 μL of isopropanol, and the absorbance was measured with a microplate reader. The measurement wavelength was 570 nm, and the reference wavelength was 620 nm.

  The cell growth rate was calculated from the absorbance, using as a control the culture with addition of the vascular endothelial cell basal medium 1 without addition of the extract of the present invention (HuMedia + FBS in FIGS. 1 to 3). The result of the hop extract is shown in FIG. 1, and the result of the marjoram extract is shown in FIG. Moreover, the result of blending in combination is shown in FIG. In the figure, the vertical axis represents the cell growth rate, the horizontal axis represents the amount (% by mass) of each extract added to the medium 2 in (ii) above, * represents p <0.05, and ** represents p. <0.01 was shown.

If the cell growth rate on the vertical axis is smaller than the culture medium 2 in (ii) above, the cell growth rate was “suppressed” or “cell growth rate was suppressed”.
From FIG. 1, it was confirmed that the hop extract suppressed excessive proliferation of HUVEC at a concentration of 0.125% by mass or more. Moreover, it confirmed that the suppression tendency was shown with respect to excessive proliferation in the density | concentration of 0.0625 mass% or more.
From FIG. 2, it was confirmed that the marjoram extract suppressed excessive proliferation of HUVEC at a concentration of 1.0% by mass or more. Moreover, it confirmed that the suppression tendency was shown with respect to excessive proliferation in the density | concentration of 0.5 mass% or more.
From FIG. 3, it was confirmed that each inhibitory action was enhanced by the combination of hops and marjoram extract. The mixture of hop extract and marjoram extract is effective by combining hop extract with a concentration of 0.0625 mass% or more and marjoram extract with a concentration of 0.125 mass% or more in combination. It was confirmed.

  Also, from FIG. 3, when using a mixture of hop extract and marjoram extract, the amount of hop extract may be larger, the amount of marjoram extract may be larger, but when the amount of marjoram extract is increased, It was confirmed that the effect was obtained with a small amount of the entire mixture. When the amount of marjoram extract is increased, it is predicted that the hop extract has the effect of suppressing the excessive proliferation of HUVEC possessed by the marjoram extract and is synergistically effective.

In addition, if the cell growth rate on the vertical axis is larger than that of the culture medium 1 described in (i) above, “no toxic tendency to cells” was set.
From FIG. 1, it was confirmed that the hop extract has no tendency to be toxic to cells at a concentration of less than 4.0% by mass.
From FIG. 2, it was confirmed that the marjoram extract had no tendency to be toxic to cells at a concentration of less than 2.0% by mass.
From FIG. 3, the mixture of hop extract and marjoram extract has a tendency to be toxic to cells at each concentration lower than the combination of 4.0% by mass of hop extract and 2.0% by mass of marjoram extract. Confirmed that there is no.

Examples 1 and 2 (skin cream)
The following components (1) to (5) were mixed and heated to dissolve to 75 ° C., and mixed, heated and melted to 75 ° C. (6) to (9) and (12) were added and emulsified, After stirring and cooling, (10) and (11) were added to obtain a skin cream.
(Composition) (mass%)
(1) Stearic acid 1.00
(2) Batyl alcohol 1.00
(3) Glycerol monostearate 0.50
(4) Squalane 15.00
(5) Sorbitan monostearate 2.00
(6) Polyoxyethylene (20) sorbitan monostearate 2.00
(7) Potassium hydroxide 0.05
(8) Carboxyvinyl polymer 0.10
(9) Methyl parahydroxybenzoate 0.10
(10) Hop extract (Example 1), Marjoram extract (Example 2) 1.00
(11) Fragrance 0.10
(12) Purified water 77.15
Total amount 100.00

<Use test of skin cream of Examples 1 and 2>
20 panelists (7 males, 29-48 years old, 13 females, 26-53 years old) who are exposed to sunlight on a daily basis and feel the weakness of their skin 20 panelists (8 men, age 27-46 years, 12 women, age 28-55 years) were tested as the second group.
As Comparative Example 1, a skin cream containing the same amount of moisturizing liquid (water / 1,3 butylene glycol = 1: 1) in place of the hop extract and marjoram extract of Examples 1 and 2 was used.
The skin cream of Examples 1 and 2 and the skin cream of Comparative Example 1 were applied to the face of the panel at a single application amount of 1.5 mg / cm ² in a blind test.
The usage period was one month from October 1 to the last day, and it was used once after every morning face wash.
The skin elasticity (Example 1 and Comparative Example 1) and sensory evaluation (Example 1 and Comparative Example 1) in Table 1 were tested with a panel 1 panel.
The skin elasticity (Example 2 and Comparative Example 1) and sensory evaluation (Example 2 and Comparative Example 1) in Table 2 were tested with a panel 2 panel.

The evaluation criteria are as follows.
(Skin elasticity)
Each panelist evaluated in 4 stages of “increased”, “no change”, “slightly reduced”, and “decreased”, and indicated by the number of people. The results are shown in Table 1.
(sensory evaluation)
Each panelist evaluated and indicated by the number of people in four stages: “I felt tension”, “No change”, “Slightly no tension”, “No tension”. The results are shown in Table 2.

  As shown in Tables 1 and 2, Examples 1 and 2 showed higher skin elasticity and a similar tendency in sensory evaluation as compared with Comparative Example 1.

As described above in detail, the topical skin preparation of the present invention suppresses angiogenesis induced as a result of an increase in VEGF that can be caused by UV exposure, and changes in skin structure caused by excessive angiogenesis, particularly elastin It was confirmed that the elasticity of the skin due to the degradation and the formation of aging skin can be prevented.
In addition, since the test concentration in Examples 1 and 2 does not show a tendency to toxicity to cells and no skin irritation is observed, it was confirmed that this is an excellent external preparation for skin with less safety concerns. .

Example 3 (Skin Cream)
In place of 1% by mass of component (10) shown in Examples 1 and 2, 3% by mass of a mixture of 1% by mass of hop extract and 2% by mass of marjoram extract is blended, and (12) 75.15% by mass is blended. A skin cream was obtained in the same manner as in Examples 1 and 2 except that.
When skin elasticity and sensory evaluation similar to those of Examples 1 and 2 were performed, effects equivalent to or higher than those of Examples 1 and 2 were confirmed.

  The external preparation for skin of the present invention can be used as cosmetics and quasi drugs for application to the skin.

Claims (5)

  One or more extracts selected from the hop (HUMULUS LUPULUS) female spikelet and the marjoram (ORIJANUM MAJORANA = MAJORANA HORTENSIS) leaf extract as an active ingredient having an angiogenesis inhibitory action of 0.05 to 5 mass % Skin external preparation.   The skin external preparation according to claim 1, wherein the extract is an extract of hops female spikelets, and the extract is contained in an amount of 0.05 to 3% by mass as an active ingredient of an angiogenesis inhibitory action.   The external preparation for skin according to claim 1, wherein the extraction part is an extract of marjoram leaves, and the extract contains 0.3 to 1.75% by mass as an active ingredient for inhibiting angiogenesis.   The extract is a mixture of an extract of hops female spikelets and an extract of leaves of marjoram, and the content ratio of the extract of hops female spikes is 0.05 to 3% by mass as an active ingredient for angiogenesis inhibitory action The external preparation for skin according to claim 1, wherein the content ratio of the extract of marjoram leaves is 0.1 to 1.5% by mass as an active ingredient for angiogenesis-inhibiting action.   The skin external preparation according to any one of claims 1 to 4, wherein the extract is obtained using a mixed solvent of water and an organic solvent selected from ethanol, 1,3-butanediol, and isopropanol. .