A composition containing dextran sulfate or its salt, royal jelly extract, and bilberry leaf extract.

Abstract

The present invention aims to provide a composition useful for inhibiting angiogenesis or vascular loss. ru. [Solution] In one embodiment of the present invention, (A) dextran sulfate or a salt thereof, (B) Prepare a composition containing royal jelly extract and (C) bilberry leaf extract. In one embodiment, in the manufacture of an external composition, dextran sulfate or its salt, rh The invention also relates to the use of compositions containing jelly extract and bilberry leaf extract.

Description

Technical Field

[0001] The present invention relates to a composition containing dextran sulfate or its salt, royal jelly extract, and bilberry leaf extract.

Background Art

[0002] Blood plays a role in delivering nutrients and oxygen to cells and collecting waste products. Blood flow insufficiency causes nutritional disorders of the skin, leading to rough and flabby skin, or uneven skin color.

[0003] The state of blood flow depends on the state of blood vessels. Therefore, strengthening and maintaining blood vessels including capillaries and preventing the disappearance of blood vessels are important for maintaining healthy skin.

Summary of the Invention

Problems to be Solved by the Invention

[0004] However, the mechanism for strengthening blood vessels and preventing the disappearance of blood vessels and the means for efficiently achieving it have not been fully studied.

[0005] One embodiment of the present invention aims to provide a composition that can surely affect blood vessels by acting on cells.

Means for Solving the Problems

[0006] As a result of intensive studies to solve the above problems, the inventors focused on the action on blood vessels via mesenchymal stem cells, and found that a composition containing dextran sulfate or its salt, royal jelly extract, and bilberry leaf extract has excellent effects, and thus completed the present invention. ​

[0007] One embodiment of the present invention is as follows: [1] (A) Dextran sulfate or its salt, (B) Royal jelly extract, and (C) Bilbe A composition containing Lee leaf extract. [2] The above-mentioned component (A) is contained in a mass of 0.0000005 to 1%, relative to the total amount of the composition, The composition described in [1]. [3] The above-mentioned component (B) is added to the total amount of the composition in an extract purity of 0.0000005 to 1 The composition according to [1] or [2], containing mass%. [4] The above-mentioned component (C) is added to the total amount of the composition in an extract purity of 0.00000005 to A composition according to any one of [1] to [3], containing 0.5% by mass. [5] Furthermore, the method described in any one of [1] to [4] contains 1,3-butylene glycol. The composition of. [6] In the manufacture of external compositions, dextran sulfate or its salt, royal jelly extract, and Use of a composition containing bilberry leaf extract. [Effects of the Invention]

[0008] According to the present invention, an excellent composition capable of acting on blood vessels can be obtained. [Brief explanation of the drawing]

[0009] [Figure 1A] Figure 1A is a graph showing the length of blood vessels 16 hours after cell seeding in the example experiment. [Figure 1B] Figure 1B is a graph showing the length of blood vessels 16 hours after cell seeding in the example experiment. [Figure 2A]Figure 2A is a graph showing the length of blood vessels 24 hours after seeding cells in the test of the example. [Figure 2B] Figure 2B is a graph showing the length of blood vessels 24 hours after seeding cells in the test of the example. [Figure 3A] Figure 3A is a graph showing the length of blood vessels 40 hours after seeding cells in the test of the example. [Figure 3B] Figure 3B is a graph showing the length of blood vessels 40 hours after seeding cells in the test of the example. [Figure 4] Figure 4 is a photograph showing the state where blood vessels disappear over time in the test of the example.

Mode for Carrying Out the Invention

[0010] [Composition containing dextran sulfate or its salt, royal jelly extract, and bilberry leaf extract The composition of the present invention is a composition containing (A) dextran sulfate or its salt, (B) royal jelly extract , and (C) bilberry leaf extract.

[0011] [((A) Dextran sulfate or its salt) The dextran sulfate used in the present invention is a substance having a structure in which a part of the hydroxyl groups of dextran is sulfated .

[0012] Dextran is, for example, a polysaccharide consisting only of glucose produced by fermenting sucrose with lactic acid bacteria or its hydrolyzate.

[0013] The molecular weight of the dextran sulfate used in the present invention is about 1,000 to 1,000 ,000 in terms of viscosity average molecular weight, and the number of SO3H groups is in the range of 1% to 30% as the sulfur content .

[0014] ​Examples of dextran sulfate salts include alkali metal salts such as sodium and potassium salts. Of these, dextran sulfate sodium salt is preferred.

[0015] Dextran sulfate or its salt is preferably dextran sulfate sodium, Sodium strane sulfate (viscosity-average molecular weight 1,000-5,000, sulfur content 3-3%) 0%) is more preferable, and dextran sulfate sodium (viscosity-average molecular weight 1,000~5, 000, sulfur content of 10-25% is more preferable, and dextran sulfate sodium ( A viscosity-average molecular weight of 1,500 to 2,700 and a sulfur content of 15 to 20% are particularly preferred.

[0016] The content of dextran sulfate or its salt in the composition may be adjusted as appropriate and is not limited to it. However, for example, it can be 0.0000005% by mass or more relative to the total amount of the composition. 0.000001 mass% or more, 0.000005 mass% or more, 0.00001 mass% or more It may also be 0.00005% by mass or more. In addition, the content of dextran sulfate or its salt may be The amount can be, for example, 1% by mass or less relative to the total amount of the composition, and 0.8% by mass or less. Examples include 0.5% by mass or less, 0.3% by mass or less, 0.1% by mass or less, etc. The content of quistran sulfate or its salt is, for example, 0.000000 of the total amount of the composition. 5~1% by mass, 0.000001~0.8% by mass, 0.000005~0.5% by mass, 0 Examples include 0.000001 to 0.3 mass%, 0.00005 to 0.1 mass%, etc.

[0017] ((B) Royal jelly extract) The royal jelly extract used in this invention is obtained by using conventionally known techniques to extract the royal jelly from the throat of a honeybee. This is a substance extracted from secretions from the head. The extraction method is not particularly limited, Water extracts are preferred. For example, by adding water or an ethanol solution to royal jelly, the result is obtained. It can also be obtained by filtering the solution. The royal jelly extract used in this invention is a commercially available product. You can also use this.

[0018] The amount of royal jelly extract in the composition may be adjusted as appropriate and is not limited to that amount. For example, the extract purity (evaporation residue) is 0.0000005 by mass relative to the total amount of the composition. It can be % or more, 0.000001% by mass or more, 0.000005% by mass or more, It may also be 0.00001% by mass or more, 0.00005% by mass or more, etc. Also, The amount of jelly extract can be, for example, 1% by mass or less of the total amount of the composition. 0.8% by mass or less, 0.5% by mass or less, 0.3% by mass or less, 0.1% by mass or less, etc. It can be expressed as follows. Furthermore, the royal jelly extract content is, for example, 0.5% of the total composition. 0000005~1% by mass, 0.000001~0.8% by mass, 0.000005~0. Possible values ​​include 5% by mass, 0.00001-0.3% by mass, and 0.00005-0.1% by mass. .

[0019] The ratio of component (A) to component (B) in the composition is 0.0 parts by mass of component (A). The amount may be 1 to 100 parts by mass, 0.05 to 50 parts by mass, or 0.1 to 10 parts by mass.

[0020] ((C) Bilberry leaf extract) The bilberry leaf extract used in this invention is obtained from the leaves of the bilberry plant, a member of the Ericaceae family, and is conventionally available This was extracted using knowledge technology. The extraction method is not particularly limited, but 1,3- A butylene glycol aqueous solution extract is preferred. For example, 1,3-butylene glycol in bilberry leaves. It can also be obtained by adding an aqueous glycol solution or an ethanol solution and filtering the resulting solution. The bilberry leaf extract used in this invention can also be a commercially available product.

[0021] The content of bilberry leaf extract in the composition may be adjusted as appropriate and is not limited to, for example. For example, the extract content (evaporation residue) is 0.00000005 by mass relative to the total amount of the composition. It can be 0% or more, 0.00000001% by mass or more, 0.0000005% by mass The above may be 0.0000001% by mass or more, 0.000005% by mass or more, etc. Furthermore, the bilberry leaf extract content should be, for example, 0.5% by mass or less of the total amount of the composition. It can be 0.3% by mass or less, 0.1% by mass or less, 0.08% by mass or less, 0.0 Examples include 5% by mass or less. In addition, the content of bilberry leaf extract may be, for example, the total amount of the composition. In relation to the quantity, 0.00000005 to 0.5 mass%, 0.0000001 to 0.3 mass% , 0.0000005~0.1% by mass, 0.000001~0.08% by mass, 0.000 Examples include 0.05 to 0.05 mass%.

[0022] The ratio of component (A) to component (C) in the composition is 0.0 parts by mass of component (A). It may be 0.1 to 10 parts by mass, 0.005 to 5 parts by mass, or 0.01 to 1 part by mass.

[0023] The composition of the present invention may contain, insofar as it does not impair the effects of the present invention, component (A), component (B), and (C) In addition to component (C), it may also contain polyhydric alcohols or nonionic surfactants. Although not, polyhydric alcohols are particularly preferred when the composition of the present invention is used as an external composition. It is used properly.

[0024] In this specification, when "polyhydric alcohol" refers to a molecule containing hydroxy(-OH) This refers to alcohols containing two or more groups. From the viewpoint of achieving the effects of the invention, polyhydric alcohols are, Divalent to pentavalent polyhydric alcohols are preferred. Examples of such alcohols include glycerin and isopentol. Ludiol, dipropylene glycol, 1,3-butylene glycol, 1,3-propane Diol, 3-methyl-1,3-butanediol, propylene glycol, pentanediol Polyethylene glycol, hexanediol, octanediol, diglycerin, or polyethylene glycol Examples include 1,3-butylene glycol, which is particularly preferred as a polyhydric alcohol. Polyhydric alcohols can be used individually or in combination of two or more types.

[0025] The total content of polyhydric alcohols in the composition may be adjusted as appropriate and is not limited to, for example If so, it can be 1% by mass or more, 3% by mass or more, or 5% by mass or more, relative to the total amount of the composition. The amount may be 8% by mass or more, 10% by mass or more, etc. Also, the total content of polyhydric alcohols is For example, it can be 50% by mass or less, 45% by mass or less, and 4% by mass of the total amount of the composition. Examples include 0% by mass or less, 35% by mass or less, 30% by mass or less, etc. Also, polyhydric alcohols. The total content is, for example, 1-50% by mass, 3-45% by mass, and 5-3% by mass, relative to the total amount of the composition. Examples include 0% by mass, 8-35% by mass, 10-30% by mass, etc.

[0026] The content of 1,3-butylene glycol in the total amount of the composition of the present invention is, for example, It can be 0.1% by mass or more relative to the total amount of the substance, 0.3% by mass or more, or 0.5% by mass. The above may be 0.8% by mass or more, 1% by mass or more, etc. Also, 1,3-butyleneglycine The total content of kohl can be, for example, 50% by mass or less of the total amount of the composition. Examples include 45% by mass or less, 40% by mass or less, 35% by mass or less, 30% by mass or less, etc. Furthermore, the total content of 1,3-butylene glycol is, for example, 0.1% of the total amount of the composition. 50% by mass, 0.3-45% by mass, 0.5-40% by mass, 0.8-35% by mass, 1-30 Examples include mass percent.

[0027] (Nonionic surfactant) Examples of nonionic surfactants include hexaglyceryl monolaurate and monomil. Hexaglyceryl stearate, hexaglyceryl monostearate, hexaglyceryl monooleate Glyceryl, Decaglyceryl monomyristate, Decaglyceryl monostearate, Mono Decaglyceryl isostearate, decaglyceryl monooleate, decaglyceryl distearate Polyglycerol fatty acid esters such as glyceryl and decaglyceryl diisostearate; Addition of 5 moles of polyoxyethylene monostearate (referred to as "POE(5)"; the same applies hereafter) Glyceryl, POE(15) glyceryl monostearate, POE(5) monooleate Polyoxyethylene glycerides such as glyceryl monooleate POE(15) glyceryl Fatty acid esters; POE(20) sorbitan monostearate, POE(6) sorbitan monostearate POE(20) sorbitan tristearate, POE(20) monoisostearate Sorbitan, POE(6) monooleate, POE(20) trioleate Polyoxyethylene sorbitan fatty acid esters such as rubitan; POE(60) sorbitol tetrastearate, POE(30) sorbitol tetraoleate Bit, POE(40) sorbitol tetraoleate, POE(60) tetraoleate Polyoxyethylene sorbitol fatty acid esters such as sorbitol; POE(10) Lanolin, POE(20) Lanolin, POE(5) Lanolin Alcohol , POE(20) sorbitol beeswax and other polyoxyethylene lanolin / lanolin alcohol Beeswax derivatives; POE(20) castor oil, POE(40) castor oil, POE(50) castor oil, POE (60) Castor oil, POE (20) Hydrogenated castor oil, POE (30) Hydrogenated castor oil, POE (40) Hydrogenated castor oil, POE(60) Hydrogenated castor oil, and other polyoxyethylene castor oils. Hydrogenated castor oils; POE(5) phytosterol, POE(10) phytosterol, POE(25) phytosterol Polyoxyethylene sterols and hydrogenated sterols such as iodistanol; POE(2) Lauryl ether, POE(4.2) Lauryl ether, POE(9) La Uryl ether, POE(5.5) cetyl ether, POE(7) cetyl ether, PO E(10) cetyl ether, POE(4) stearyl ether, POE(7) oleyl ether - ether, POE(10) oleyl ether, POE(10) behenyl ether, POE( 2) (C12-15) alkyl ether, POE(4)(C12-15) alkyl ether Lu, POE(5) secondary alkyl ether, POE(7) secondary alkyl ether, POE( 9) Polyoxyethylene alkyl ethers such as alkyl ethers and POE(12) alkyl ethers Luethers; Addition of 1 mole of polyoxypropylene (referred to as "POP(1)" below) POP(4) Chill ether, POE(10)POP(4) Cetyl ether, POE(20)POP(8 ) Cetyl ether, POE(20)POP(6) Decyltetradecyl ether, POE( 30) POP(6) Decyltetradecyl ether and other polyoxyethylene polyoxypropyl Pyrene alkyl ethers. Addition of 10 moles of polyethylene glycol monolaurate (" It is written as "PEG(10)" (the same applies below), PEG(10) monostearate, monooleic acid PEG(10), PEG diisostearate, and other polyethylene glycol fatty acid esters kind; PEG(8) glyceryl isostearate, PEG(10) glyceryl isostearate PEG-15 glyceryl isostearate, PEG-20 glyceryl isostearate Ceryl, PEG(25) Glyceryl Isostearate, PEG Glyceryl Isostearate Polyoxyethylene glyceryl isostearates such as (30); Diglyceryl monostearate, decaglyceryl decastearate, decaglyceryl de Caisostearate, diglyceryl diisostearate, and polyglyceryl monolaurate Polyglycerol fatty acid esters such as Lyl Examples are given.

[0028] (water) The composition of the present invention may also contain water. Total content of water relative to the total amount of the composition of the present invention is 1% by mass or more, 10% by mass or more, 20% by mass or more, 30% by mass or more, 40% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more, 80% by mass or more, or 90% by mass or more It can be set to 99% by mass or less, 90% by mass or less, 80% by mass or less, 70% by mass or less The following can be set: 60% by mass or less, 50% by mass or less, and 40% by mass or less.

[0029] When the composition of the present invention is a liquid composition, the pH can be 4 or higher, and 4.5 or higher. The above can be 5 or more, etc. Also, it can be 10 or less, 9 or less, 8 or less, 7 or less Lower grades are acceptable. For example, grades 4-9, 4.5-8, 5-7, etc., are also acceptable.

[0030] (Additional ingredients) The composition of the present invention may also contain components other than those mentioned above, as long as the effects of the present invention are not impaired. It can be used. Additional ingredients include various plant extracts, primary alcohols, pH adjusters, and antioxidants. Examples include agents, thickeners, inorganic salts, stabilizers, irritation reducers, colorants, or fragrances. You may use one or more of these.

[0031] [Application] The composition of the present invention is derived from mesenchymal stem cells (also known as "mesenchymal stromal cells" or "MSCs"). It acts on mesenchymal stem cells, promoting the proliferation of vascular endothelial cells and stimulating angiogenesis. This can promote and suppress vascular loss.

[0032] Here, promoting angiogenesis mainly refers to the creation of new capillaries and the lengthening of blood vessels. This refers to either of the following. Vascular loss suppression means preventing the loss or regression of existing blood vessels, or preventing their loss. It means extending the time spent there.

[0033] Here, mesenchymal stem cells are not limited to any specific type, but rather any mesenchymal stem cells that may exist in the body. It is possible, but it is preferable that it be of adipose-derived or bone marrow-derived mesenchymal stem cells ("adipose-derived mesenchymal stem cells"). It is more preferable that the cells are derived from mesenchymal stromal cells, also known as "AD-MSCs."

[0034] In one embodiment, the composition of the present invention provides vascular endothelial growth factor (V) in mesenchymal stem cells. By promoting the production of EGF and / or basic fibroblast growth factor (bFGF) This can increase the proliferation of vascular endothelial cells, promote angiogenesis, and suppress vascular loss. Therefore, vascular endothelial growth factor (VEGF) and / or basic fibroblast growth factor (bFGF) To promote the production of means increasing the gene expression of VEGF and / or bFGF, and This refers to increasing the production of vegetarian and / or vegetarian and / or bFGF.

[0035] The composition of the present invention prevents skin thinning or improves thinned skin; improves skin complexion or color To improve skin texture; to improve skin firmness and radiance; to improve the facial contour; or to reduce wrinkles and blemishes. It can improve the smoothness.

[0036] In one embodiment, the composition of the present invention may be used for purposes other than food and beverages. It can be used.

[0037] In one embodiment, the composition of the present invention is suitably used as a topical composition.

[0038] When used as a so-called external composition such as a pharmaceutical, quasi-drug, or cosmetic, the present invention The composition is preferably a skincare cosmetic (e.g., lotion, cream, facial cream). (Facial lotion, emulsion, mask, serum, face wash, cleansing product), makeup cap Cosmetics (e.g., eyeliner, eyebrow products), scalp and hair cosmetics (e.g., scalp lotion) Examples include lotions, scalp creams, and more preferably, skincare cosmetics (e.g., (Lotion, cream, face cream, face lotion, emulsion, mask, serum) It is possible, but it is not limited to these.

[0039] Alternatively, but not limited to, in one embodiment, the composition of the present invention may be applied to areas other than the scalp. It can be used as a topical composition for application to [the subject].

[0040] Alternatively, but not limited to, in one embodiment, the composition of the present invention is a hair growth agent. It can be used as an external composition.

[0041] Thus, when used in so-called external compositions such as pharmaceuticals, quasi-drugs, and cosmetics, The dosage forms of the composition of the present invention are liquid, suspension, cream, lotion, and aerosol. Other examples include:

[0042] The composition of the present invention is administered once to several times a day, usually 1 to 3 times a day, 1 to 2 times a day, or as needed. It can be applied over any period and interval.

[0043] [Contains dextran sulfate or its salt, royal jelly extract, and bilberry leaf extract] [Culture medium for mesenchymal stem cells] The present invention also relates to (A) dextran sulfate or a salt thereof, (B) royal jelly extract, and (C) This relates to a culture medium for mesenchymal stem cells containing bilberry leaf extract.

[0044] Other culture medium components used for maintaining or promoting mesenchymal stem cells contained in the culture medium, For the culture medium components used in animal cell culture, the components included in general basal culture media are used. In addition, components (A), (B), and (C) can be added to this. Alternatively, commercially available culture media for various stem cells can be used. Examples include Eagle's Minimum Essential Medium (MEM) and Dulbecco's Modified Eagle Medium (DMEM). ), α-Eagle Minimum Essential Medium (αMEM), Iscove Modified Dulbecco's Medium (IMDM) ), F-12 medium, F-10 medium, DMEM / F12 medium, RPMI-1640 medium, between Leaf cell basal medium (MSCBM), E8 (Essential 8) medium, TeSR-E 8 medium, mTeSR1 medium, R:STEM medium (manufactured by Rohto Pharmaceutical Co., Ltd.) and mixtures thereof Examples include composite culture media. The amounts of component (A), component (B), and component (C) are [(A) Dextrin (B) Strain sulfate or its salt, royal jelly extract, and (C) bilberry leaf extract The conditions described in [Compositions containing] are as follows.

[0045] In the culture medium for mesenchymal stem cells, (A) dextran sulfate or its salt, (B) royal jelly - By combining the extract and (C) bilberry leaf extract, the culture of mesenchymal stem cells Components useful for inhibiting angiogenesis and vascular loss can be obtained from the supernatant. This includes, but is not limited to, vascular endothelial growth factor (VEGF) and / or basic fibroblasts. One example is follicular growth factor (bFGF).

[0046] [Contains dextran sulfate or its salt, royal jelly extract, and bilberry leaf extract] [Method for using a composition containing mesenchymal stem cells in the culture of mesenchymal stem cells] This invention relates to dextran sulfate or its salts, royal jelly extract, and bilberry leaf extract. The present invention also relates to a method for using a composition containing the exudate in the culture of mesenchymal stem cells. (A) component, (B The amounts of (A) dextran sulfate or its salt, (B) royal In accordance with the conditions described in [Composition containing jelly extract and (C) bilberry leaf extract] The basal culture medium also contains [dextran sulfate or its salt, royal jelly extract, and bicarbonate]. This is the same as described in "[Culture medium for mesenchymal stem cells containing berry leaf extract]".

[0047] [Contains dextran sulfate or its salt, royal jelly extract, and bilberry leaf extract] By using the composition in the culture of mesenchymal stem cells, angiogenesis by mesenchymal stem cells is achieved. Methods to improve the ability to suppress vasodilation and vascular loss. This invention relates to dextran sulfate or its salts, royal jelly extract, and bilberry leaf extract. By using a composition containing the exudate in the culture of mesenchymal stem cells, This also relates to methods for improving angiogenesis and suppression of vascular loss. (A) component, (B) component, The amounts of (A) dextran sulfate or its salt, (B) royal jelly extract The conditions described in [Composition containing (C) bilberry leaf extract] shall apply. The basal medium consists of [dextran sulfate or its salt, royal jelly extract, and bilberry]. This is the same as described in [Culture medium for mesenchymal stem cells containing leaf extract]. [Examples]

[0048] Next, the present invention will be specifically described with reference to examples, but the present invention is limited to the following examples. It's not that. The raw materials used in the following tests are as follows: Dextran sulfate sodium (viscosity-average molecular weight 1,500-2,700, sulfur content) 15-20% (Manufactured by Meito Sangyo Co., Ltd.) Royal jelly extract-containing solution (extract 1.25% by mass, water 48.75% by mass, 1 ,3-butylene glycol 50% by mass: 1,3-butylene glycol in an aqueous extract of royal jelly Equal amounts of lenglycol added (manufactured by Ichimaru Falcos Co., Ltd.) Bilberry leaf extract containing solution (extract 0.2% by mass, water 49.9% by mass, 1,3- Butylene glycol 49.9% by mass: 50% by mass of bilberry leaves 1,3-butylene glycol Recall aqueous solution extract (manufactured by Ichimaru Falcos Co., Ltd.)

[0049] [Test Example 1] (Preparation of supernatant from adipose-derived mesenchymal cell cultures) Cells isolated from human fat (LIFE LINE, FC-0034) are used in adipose-derived mesenchymal cells. They were used as stem cells (AD-MSCs). AD-MSCs are specially prepared for hMSC culture. AD-MSC was cultured using a culture medium (R:STEM medium, manufactured by Rohto Pharmaceutical Co., Ltd.). 9.5x10 4 / well / 6-well plate (Corning cell bind) Seeds were sown in 3335), and the following day, dextran sulfate sodium, royal jelly extract, and With or without the addition of a three-component mixture of bilberry leaf extract (extract complex). The culture supernatant was then collected after 48 hours of incubation. The culture was carried out at 37°C in 5% CO2. The process is carried out in an incubator, and the supernatant is collected by centrifugation at 800g for 10 minutes at 4°C. went.

[0050] (Preparation of vascular endothelial cells) As vascular cells, normal umbilical vein endothelial cells We used Endothelial Cells (HUVEC). HUVEC is, Purchased cells (HUVECs (LIFE LINE, FC-0044)) were used.

[0051] (Confirmation test of the disappearance of compositions for vascular-like structure formation and vascular endothelial function improvement using Matrigel) Base containing laminin, collagen IV, entactin, and heparin sulfate proteoglycan Matrigel containing membrane extract (Geltrex® LDEV-Free Red uced Growth Factor Basement Membrane Mat Add 40 μl of rix (Gibco, A1413201) to each well and store at 37°C. Then, leave it in a 5% CO2 incubator for more than 30 minutes, and then place the gel-filled 96-well plate in the incubator. A well was prepared. 11,000 vascular endothelial cells (HUVECs) were seeded into each well. I planted the seeds.

[0052] At this time, R:STEM medium (manufactured by Rohto Pharmaceutical Co., Ltd.) was used as the basal culture medium, and the medium was used , the culture supernatant of AD-MSCs treated with extract complex or the extract complex 120 μl each of the culture supernatant from untreated AD-MSCs, or extract complex We added only sucrose and examined its effects on angiogenesis and vascular loss.

[0053] The following were used as the exponential complex. Excomplex (Figure 1A, Figure 2A, Figure 3A): Dextran sulfate sodium 0.0 0.01%, Royal Jelly Extract 0.00125%, Bilberry Leaf Extract 0.0002% Excomplex (Figures 1B, 2B, 3B): Dextran sulfate sodium 0.0 0.001%, Royal Jelly Extract 0.000125%, Bilberry Leaf Extract 0.0000 2%

[0054] Verification of angiogenesis and angiogenesis was performed using the ImageXpress Confo imaging system. Images were taken using cal (Molecular Devices Inc.) to identify cells and seeding of each component. At the time of sowing, 16 hours after sowing, 24 hours after sowing, and 40 hours after sowing The process was carried out. After each time interval, the total length of the blood vessels was analyzed using ImageXpress analysis on the acquired images. It was calculated using a program.

[0055] (result) The average results of four tests or eight tests are shown in Figures 1A, 2A, and 3A. The average results of 6 or 8 tests are shown in Figures 1B, 2B, and 3B. Figures 1A, 2A, and 3A show the extract complex as bilberry leaf Extract 0.0002%, royal jelly extract 0.00125%, and dextran sulfur These are the results for a sodium phosphate concentration of 0.001%. Figures 1B, 2B, and 3B are also shown. , bilberry leaf extract 0.00002%, royal jelly extract 0.000125%, The results are for concentrations of 0.0001% sodium dextran sulfate. See Figures 1A and 1B. Based on the length of blood vessels 16 hours after seeding, extract complex and AD-MSC supernatant were added. In this case, it was found that angiogenesis was promoted compared to the culture medium alone. In AD-MSC supernatant treated with condensate, significant angiogenesis was observed. It is expected that, within the body, it will act on blood vessels through cells, promoting angiogenesis. (Figure) Based on the length of blood vessels 24 hours after seeding in 2A and B, the excomplex and AD- When MSC supernatant was added, the degree of vascular disappearance was milder compared to when only the culture medium was added. It was found to suppress it. This was especially true in AD-MSC supernatant treated with Excomplex. Significant suppression of vascular loss was observed. Figures 3A and 3B show the results 40 hours after seeding. When the extract complex and AD-MSC supernatant are added, compared to the culture medium alone, the blood vessel length is measured. In comparison, it was found to have an effect of suppressing vascular loss. A treated with Excomplex This was particularly pronounced in the D-MSC supernatant. This suggests that, within the body, it acts on blood vessels through cells. This indicates that it is effective in suppressing vascular loss and maintaining blood vessels.

[0056] Furthermore, Figure 4 shows the state of vascular disappearance from 16 hours to 40 hours, from the top to the bottom. The images are shown below. Here, the left image shows the state with only the culture medium, and the center image shows AD-M The image on the right shows the result after applying SC supernatant, and the image on the right shows AD-MS processed with Excomplex. This is the state after applying the C supernatant.

[0057] Thus, the composition of the present invention exhibits excellent effects in inhibiting angiogenesis and vascular loss. It was revealed that...

Claims

1. A topical composition containing (A) dextran sulfate or a salt thereof, (B) an aqueous extract of royal jelly, and (C) an aqueous 1,3-butylene glycol extract of bilberry leaves.

2. The topical composition according to claim 1, wherein component (A) is contained in 0.0000005 to 1% by mass of the total amount of the composition.

3. The topical composition according to claim 1, wherein the above-mentioned component (B) is contained in an extract purity of 0.0000005 to 1% by mass with respect to the total amount of the composition.

4. The topical composition according to claim 1, wherein the above-mentioned component (C) is contained in an extract purity of 0.00000005 to 0.5% by mass with respect to the total amount of the composition.

5. The topical composition according to claim 1, for use on the face.

6. Furthermore, the topical composition according to any one of claims 1 to 5, further containing 1,3-butylene glycol.

Images