Composition containing Nemunoki extract

Abstract

Provided are: a composition for suppressing or preventing disturbances of the expression rhythm of the clock gene; a composition for preventing erythema of the skin induced by ultraviolet light and the like; and a composition for preventing generation of glycated products by the reaction of a protein and a sugar.　The present invention is a composition for suppressing or preventing disturbances of the expression rhythm of the clock gene, an anti-inflammatory agent, and an anti-glycation agent which contain an Albizia julibrissin extract.

Description

[Technical field]

[0001] The present invention relates to a composition containing an extract of Albizia julibrissin, which can suppress or prevent abnormalities in the expression rhythm of clock genes in skin cells that exhibit an irregular or abnormal state of expression rhythm of clock genes compared to normal expression rhythm of clock genes. The present invention also relates to an anti-inflammatory agent containing an extract of Albizia julibrissin bark as an active ingredient. More specifically, the present invention relates to a topical skin composition that uses the anti-inflammatory agent to suppress erythema after ultraviolet irradiation. The present invention also relates to an anti-glycation agent that utilizes the anti-glycation effect of an extract of Albizia julibrissin bark. More specifically, the present invention relates to an anti-glycation agent that has the effect of suppressing the production of glycation products caused by the reaction of proteins with sugars, and a topical skin composition containing the anti-glycation agent. [Background technology]

[0002] Most living organisms on Earth control their physiological functions (sleep / wake, thermoregulation, feeding, excretion, hormone secretion, etc.) in a roughly 24-hour cycle in accordance with the external environment (mainly the light / dark cycle), and this type of biological diurnal variation is called "circadian rhythm." In mammals in particular, the center of circadian rhythm function is the suprachiasmatic nucleus in the hypothalamus in the brain. Periodic signals sent from the suprachiasmatic nucleus are transmitted to peripheral tissues via neurotransmission and hormones, so that the various physiological functions of the entire body are regularly regulated with a certain cycle.

[0003] On the other hand, it has been revealed that peripheral tissues also have mechanisms for forming autonomous circadian rhythms. At the cellular level in peripheral tissues, circadian rhythms are regulated at the transcriptional level by the interaction of multiple clock genes. At the center of this is a core loop of transcription centered on the transcription factors Bmal1 and Clock. In the nucleus, Bmal1 and Clock form heterodimers, which bind to E-box sequences on DNA and regulate the expression of clock genes such as Per and cry, as well as non-clock genes such as many hormones, cytokines, and enzymes. Translated Per and Cry proteins form Per / Cry heterodimers and migrate into the nucleus, suppressing the transcriptional activity of Bmal1 / Clock. This terminates the transcription of Per and Cry genes. At the same time, PER and CRY proteins are rapidly degraded by phosphorylation and the subsequent ubiquitin-proteasome system. As a result, this negative feedback loop is released and transcriptional regulation by Bmal1 / Clock begins again. These mechanisms are carried out in a 24-hour cycle, allowing the body to keep track of the circadian rhythm.

[0004] Skin cells are no exception, and it has been suggested that aquaporins are under the expression control of BMAL1 / CLOCK with regard to barrier function. There are also reports suggesting a close relationship between reactive oxygen production and BMAL1 (Non-Patent Document 1). On the other hand, there are also reports that exposure to oxidative stress in cultured skin cells causes abnormalities in the expression rhythm of clock genes, and it is not difficult to imagine that abnormalities in the expression rhythm of clock genes in these skin cells could cause physiological dysfunction.

[0005] Thus, circadian rhythm is controlled by the expression of clock genes. Therefore, a search is being made for substances that can change the circadian rhythm by changing the expression. Patent Document 1 reports a clock gene expression regulator containing a sphingoid base as an active ingredient. In this invention, clock gene disorder is induced under a state of stress caused by hydrogen peroxide, and attention is paid to the recovery of the amplitude. Patent Document 2 reports a circadian rhythm improver containing a passionflower extract (excluding those containing harmine). In this invention, attention is paid to the enhancement of clock gene expression (amplitude) under non-stress conditions. Patent Document 3 reports a composition for promoting clock gene expression containing black ginger (Kaempferia parviflora). In this invention, attention is also paid to the enhancement of clock gene expression (amplitude) under non-stress conditions.

[0006] Searching for ingredients that act safely and effectively on clock genes in vivo is important not only from the viewpoint of providing safer and more effective ingredients to the market, but also from the viewpoint of diversifying formulations, expanding options for prescription combinations, and synergistic effects by combining different ingredients, and it goes without saying that the development of new ingredients that act on clock genes is required. In particular, there is a demand for ingredients that suppress or prevent abnormalities in clock genes induced by stress, rather than simply affecting the expression of clock genes. While there are materials on the market that focus on the amplitude of clock genes, there are few materials that focus on the phase of clock genes, and there is a demand for materials that can respond to phase changes.

[0007] In this context, Patent Document 4 found that an aqueous extract of Albizia julibrissin bark, when added to mouse embryonic fibroblasts, can suppress the gradual attenuation of the amplitude of the expression rhythm of the clock gene (Per2 gene) in the cells. Furthermore, it has been shown that the extract can advance or delay the phase of the expression rhythm of the Per2 gene in a concentration-dependent manner depending on the timing of administration to the cells, suggesting a flexible effect on clock genes.

[0008] Silk trees are deciduous tall trees of the legume family that grow naturally in sunny wetlands such as forest edges and fields in mountainous areas south of the Tohoku region in Japan, and are also distributed and cultivated overseas in China, Southeast Asia, etc. The bark is collected during the flowering season, washed with water, dried in the sun, and chopped into appropriate lengths, which is called silk bark. It is taken by the public for arthritis, lower back pain, diuretic, edema, and tonic effects, and has also been used externally as a cold compress for swelling, bruises, and joint pain with a decoction, or as a bath additive. The bark is known to contain triterpene saponins (many julibrosides), flavonoids (gerardone, isoocanin, luteolin, etc.), tannins, etc. (Non-Patent Document 2)

[0009] In recent years, the effects of UV rays on the skin and their mechanism of action have been actively studied. Usually, UV rays that reach the earth's surface are divided into UVA (320-400 nm) and UVB (290-320 nm). Generally, UVB rays are small, accounting for about 10% of the UV rays that reach us from the sun, but they have a strong effect on the skin, causing sunburn (an inflammatory reaction caused by sunburn) in which the skin turns red even in a short period of time, and suntan (a pigmentation reaction) in which the skin turns dark after a few days. UVB rays, which have a short wavelength, are known to have a strong effect on living organisms, not only causing inflammation and blemishes, but also damaging epidermal cells and DNA on the surface of the skin.

[0010] When skin tones are divided into four stages (I-IV), if an average Japanese person with type II sunbathes for about 20 minutes in Tokyo Bay in midsummer, the slight redness that appears after 12 to 24 hours is called erythema. The minimum amount of energy that causes this erythema is called the Minimal Erythema Dose (MED), and the MED for Japanese people due to UVB exposure is 0.04 to 0.07 J / m. 2 It is said that (Non-Patent Document 3).

[0011] Various anti-inflammatory agents have been proposed for the purpose of preventing and suppressing erythema, which is an inflammation induced by ultraviolet rays. Patent Document 5 reports a skin preparation for external use that is characterized by containing a plant selected from Tussilago farfara L. and Artemisia capillan's Thunb. or an extract thereof. Patent Document 6 reports a skin preparation for external use that contains ascorbic acid and its derivatives or salts thereof, glycyrrhizic acid and its derivatives or salts thereof, a component derived from rice bran, and gentiana extract. Patent Document 7 reports a skin preparation for external use that contains an apple extract.

[0012] On the other hand, it goes without saying that there is a demand for the development of new erythema suppressants from the standpoints of diversifying formulations, expanding options for formulation combinations, synergistic effects using erythema suppressants with different mechanisms, and erythema suppressants that are safer and more effective.

[0013] Furthermore, when proteins and reducing sugars are mixed and heated, the amino groups of the proteins and the carbonyl groups of the reducing sugars bind nonenzymatically to form glycation products. This reaction is called the Maillard reaction or glycation reaction. This binding reaction between proteins and reducing sugars also occurs in the body, and glycated proteins are constantly formed.

[0014] These glycated proteins ultimately form irreversible compounds called advanced glycation end-products (AGEs). AGEs are produced not only from glucose, but also from various sugars such as glucose autoxidation and decomposition products. Known AGEs include pentosidine, crosslin, carboxymethyllysine, and pyrazine.

[0015] It has become clear in recent years that AGEs accumulate with age and are closely related to skin aging. When glycation occurs in skin proteins, the amino group of lysine residues or the guanidyl group of arginine residues in proteins reacts nonenzymatically with the carbonyl group of sugar to produce AGEs, which crosslink proteins. When crosslinked structures are formed, the molecules become hard and the skin loses its inherent elasticity. In addition, the skin recognizes the crosslinked material as a foreign body, and the secretion of proteolytic enzymes increases. As a result, the skin loses firmness and elasticity, becomes brittle, and wrinkles, sagging, and dullness occur. Therefore, inhibiting protein glycation is thought to be effective in preventing and treating these symptoms.

[0016] In light of this background, research is being conducted into the use of natural products as ingredients that inhibit the glycation reaction of proteins, including AGE production inhibition. For example, Patent Document 8 describes an AGE production inhibition effect that contains Budreja axillaris leaf extract as an active ingredient. Patent Document 9 describes a protein anti-glycation agent that contains propolis or a processed product thereof as an active ingredient. Patent Document 10 describes an anti-glycation cosmetic that contains plum vinegar.

[0017] On the other hand, there is a demand for the development of new anti-glycation agents from the viewpoints of diversifying formulations, expanding options for prescription combinations, synergistic effects using anti-glycation agents with different mechanisms, and safer and more effective anti-glycation actions. [Prior art documents] [Patent documents]

[0018] [Patent Document 1] JP 2018-138524 A [Patent Document 2] JP 2018-43970 A [Patent Document 3] JP 2016-210703 A [Patent Document 4] JP 2018-058783 A [Patent Document 5] Japanese Patent Application Publication No. 8-175958 [Patent Document 6] JP 2010-47535 A [Patent Document 7] JP 2013-095704 A [Patent Document 8] JP 2011-102270 A [Patent Document 9] JP 2012-077042 A [Patent Document 10] JP 2012-214434 A [Non-patent literature]

[0019] [Non-Patent Document 1] COSMETIC STAGE Vol12,No.3 2018 [Non-Patent Document 2] Wakanyaku No.759 (2016.8) [Non-Patent Document 3] Pharmaceutical Journal, Vol.126, p677-693, 2006 Summary of the Invention [Problem to be solved by the invention]

[0020] In view of the above background, an objective of the present invention is to find a material that suppresses or prevents disturbances in the expression rhythm of clock genes. Furthermore, an objective of the present invention is to prevent various diseases and symptoms caused by disturbances in the expression rhythm of clock genes. Another objective of the present invention is to find an anti-inflammatory agent that suppresses erythema induced by ultraviolet rays. Furthermore, an objective of the present invention is to provide an anti-glycation agent that has the effect of suppressing the production of glycation products caused by the reaction of proteins and sugars. [Means for solving the problem]

[0021] In view of the above circumstances, the present inventors have discovered that an extract of Albizia julibrissin has the effect of effectively suppressing or preventing disruption of the expression rhythm of clock genes caused by oxidative stress, that an extract of Albizia julibrissin bark has an anti-inflammatory effect of suppressing erythema induced by ultraviolet light, and that an extract of Albizia julibrissin bark has an anti-glycation effect of suppressing the glycation reaction of glycation products produced by the reaction of proteins with sugar, and have thus completed the present invention.

[0022] That is, the outline of the present invention is as follows. Section 1. A composition containing an extract of Albizia julibrissin, which is used on skin cells that exhibit an irregular or abnormal rhythm of clock gene expression compared to the normal rhythm of clock gene expression. Section 2. Item 2. A composition containing an extract of Albizia julibrissin according to Item 1, which is a composition for suppressing or preventing abnormalities in the expression rhythm of clock genes. Section 3. Item 3. The composition containing an extract of Albizia julibrissin according to Item 1 or 2, wherein the irregular or abnormal expression rhythm of the clock gene is caused by any factor selected from the group consisting of oxidative stress, ultraviolet light, external stimuli due to drugs, physiological stress, and internal factors due to aging. Section 4. Item 4. The composition containing an extract of Albizia julibrissin according to Item 3, wherein the irregular or abnormal expression rhythm of the clock gene is a phase delay. Section 5. Item 5. The composition containing an extract of Albizia julibrissin according to any one of items 1 to 4, wherein the skin cells are human epidermal keratinocytes. Section 6. Item 6. The composition containing an extract of Albizia julibrissin according to any one of Items 1 to 5, wherein the clock gene is a gene that exhibits circadian rhythm in its expression rhythm. Section 7. Item 7. The composition containing an extract of Albizia julibrissin according to Item 6, wherein the clock gene is at least one gene selected from the group consisting of a Per gene, a Bmal gene, a Clock gene, a Cry gene, a Tim gene, an Npas gene, an Nr1d1 gene, and a Dbp gene. Section 8. Item 8. The composition containing an extract of Albizia julibrissin according to Item 7, wherein the clock gene is the Bmal1 gene. Section 9. Item 9. The composition according to any one of items 1 to 8, wherein the Albizia julibrissin extract is an extract of Albizia julibrissin bark. Section 10. Item 10. The composition containing an extract of Albizia julibrissin according to any one of Items 1 to 9, wherein the extract of Albizia julibrissin is present in an amount of 0.001 to 10% by mass of the total composition to be applied to the skin cells. Section 11. Item 11. The composition according to item 10, wherein the Albizia julibrissin extract is 0.005 to 1% by mass of the total composition to be applied to the skin cells. Section 12. Item 12. A cosmetic comprising the composition containing an extract of Albizia julibrissin according to any one of items 1 to 11. Section 13. Item 12. A skin external preparation comprising the composition containing an extract of Albizia julibrissin according to any one of items 1 to 11. Section 14. Item 14. The skin external preparation according to item 13, wherein the Albizia julibrissin extract is contained in an amount of 0.00001 to 5.0% by dry weight. Section 15. An anti-inflammatory agent comprising an extract of Silk tree bark obtained by extracting Silk tree bark with water. Section 16. Item 16. The anti-inflammatory agent according to item 15, which has an effect of suppressing skin erythema. Section 17. Item 17. The anti-inflammatory agent according to item 15 or 16, which has an effect of suppressing skin erythema induced by ultraviolet irradiation. Section 18. Item 18. The anti-inflammatory agent according to any one of Items 15 to 17, wherein the concentration of the Albizia julibrissin bark extract is in the range of 0.00001 to 5.0% by weight on a dry basis. Section 19. Item 19. The anti-inflammatory agent according to Item 18, wherein the concentration of the Albizia julibrissin bark extract is in the range of 0.00005 to 3.0% by weight on a dry basis. Section 20. Item 19. The anti-inflammatory agent according to Item 18, wherein the concentration of the Albizia julibrissin bark extract is in the range of 0.0001 to 1.0% by dry weight. Section 21. 21. A composition for external use on skin, comprising the anti-inflammatory agent according to any one of items 15 to 20. Section 22. An anti-glycation agent comprising an extract of Albizia julibrissin bark. Section 23. Item 23. The anti-glycation agent according to Item 22, which has an effect of suppressing the production of glycation products caused by the reaction of proteins with sugars. Section 24. Item 24. The anti-glycation agent according to Item 22 or 23, wherein the Albizia julibrissin bark extract is extracted from Albizia julibrissin bark with water. Section 25. Item 25. The anti-glycation agent according to any one of Items 22 to 24, wherein the Albizia julibrissin bark extract is contained in a concentration range of 0.00001 to 5.0% by dry weight. Section 26. Item 26. The anti-glycation agent according to Item 25, wherein the Albizia julibrissin bark extract is contained in a concentration range of 0.00005 to 3.0% by dry weight. Section 27. Item 26. The anti-glycation agent according to Item 25, wherein the Albizia julibrissin bark extract is contained in a concentration range of 0.0001 to 1.0 wt % on a dry weight basis. Section 28. Item 28. A composition for external use on skin, comprising the anti-glycation agent according to any one of Items 22 to 27. Effect of the Invention

[0023] By using the Albizia julibrissin extract of the present invention, it is expected that the disturbance of clock genes caused by oxidative stress, etc. can be suppressed or prevented, and a skin topical preparation having this effect can be provided. In addition, by using the Albizia julibrissin bark extract, it is expected that an anti-inflammatory agent that suppresses skin erythema induced by ultraviolet light can be provided, and the production of glycation products caused by the reaction of protein and sugar can be suppressed, and a skin topical composition having these effects can be provided. [Brief description of the drawings]

[0024] [Figure 1] FIG. 1 shows the results of evaluating the inhibitory or preventive effect on disruption of the expression rhythm of target clock genes in normal neonatal human epidermal keratinocytes in Example 1 (group with no hydrogen peroxide added / group with no Albizia julibrissin extract added). [Diagram 2] FIG. 2 shows the results of evaluating the inhibitory or preventive effect on disruption of the expression rhythm of target clock genes in normal neonatal human epidermal keratinocytes in Example 1 (hydrogen peroxide-added group / silvus julibrissin extract-free group). [Diagram 3] FIG. 2 shows the results of evaluating the inhibitory or preventive effect on disruption of the expression rhythm of target clock genes in normal neonatal human epidermal keratinocytes in Example 1 (hydrogen peroxide-added group / Albizia julibrissin extract-added group). [Figure 4] FIG. 11 is a graph showing a comparison of the results of skin color measurement between a test sample application group and a non-application group in Example 3. [Diagram 5] FIG. 13 shows a comparison of the measurement results of the concentration of carbonyl groups generated by saccharification reaction in a test sample-added group and a control-added group in Example 5. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] The present invention provides a composition containing an extract of Albizia julibrissin, which is characterized by suppressing or preventing abnormalities in the expression rhythm of clock genes in skin cells that show irregular or abnormal expression rhythms of clock genes compared to normal expression rhythms of clock genes. Examples of skin cells to which the composition can be applied include human epidermal keratinocytes, human dermal fibroblasts, human epidermal melanocytes, human dermal microvascular endothelial cells, human dermal microlymphatic endothelial cells, human hair papilla cells, etc. It is particularly preferable to use the composition on human epidermal keratinocytes.

[0026] An example of an extraction method for the Albizia julibrissin extract used in the present invention is shown below. The part of the plant body used in the extract is mainly the bark, but the whole plant body can also be used without being limited to this. It goes without saying that the method is not limited to the example described below.

[0027] The term "silvus julibrissin extract" refers to the plant itself, its cut pieces, dried powder, and extracts thereof. The term "extract" includes both the plant body of the genus Silvus and the plant body from which the extracting solvent has been removed, as well as the plant body that has been subjected to further purification treatment after the removal of the extracting solvent.

[0028] The Albizia extract is obtained by extracting the plant body of the Albizia genus plant, typically its branches and leaves, raw or dried and crushed, and then extracting it using water, preferably hot water at 70 to 100 ° C., or an organic solvent as an extraction solvent. Extraction can be performed by combining water extraction and organic solvent extraction as necessary, but considering the effects when applied to a living body, it is preferable to perform extraction only with water. For example, water; alcohols such as methanol, ethanol, propanol, and butanol; polyhydric alcohols such as propylene glycol and butylene glycol; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; linear and cyclic ethers such as tetrahydrofuran and diethyl ether; polyethers such as polyethylene glycol; hydrocarbons such as hexane, cyclohexane, and petroleum ether; aromatic hydrocarbons such as benzene and toluene; pyridines; oils and fats, waxes, and other oils. Among these, water, alcohols, and water-alcohol mixtures are preferred, and hot water is particularly preferred as the extraction solvent.

[0029] In the case of hot water extraction, typically, the plant body of the plant of the genus Albizia is extracted under heating and reflux. The extraction conditions vary depending on the solvent used. For example, when extracting with water, it is preferable to use 1 to 100 parts by weight of water per part by weight of Albizia, extract at a temperature of 4 to 100°C for 10 minutes to 7 days, and more preferable to use 5 to 20 parts by weight of water per part by weight of Albizia bark, extract at a temperature of 70 to 100°C for 30 minutes to 2 hours. In addition, from the viewpoint of further increasing the extraction efficiency, pressurization, stirring, ultrasonic treatment, etc. may be performed in conjunction with the extraction treatment. Furthermore, after such extraction treatment, it may be separated from the residue by filtration or centrifugation.

[0030] When the extract of Albizia julibrissin is distilled off under reduced pressure, it usually gives a tarry, dark brown extract, and excipients can be added as necessary. When the extract is non-toxic, such as water, it can be used in preparations as it is, but the extract from which the extract is separated can also be used in preparations. The extract from which the extract is separated can also be redissolved in a suitable solvent, such as water, and used in preparations. The extract thus obtained can also be purified or its titer improved by purification means, such as fractional extraction, partition between two solvents, fractional adsorption / elution with a suitable adsorbent, or chromatography.

[0031] The Albizia julibrissin extract obtained as described above can be used as it is to suppress or prevent abnormalities in the expression rhythm of total genes, but it is preferable to use it by blending it with an external preparation, especially an external preparation for skin. In this case, the amount to be blended with the external preparation is determined depending on the degree of absorption, the degree of action, the product form, the frequency of use, etc., and is not particularly limited, but it is desirable to make the concentration range of 0.00001 to 5.0 wt% by dry weight, preferably 0.00005 to 3.0 wt%, and particularly preferably 0.0001 to 1.0 wt%. If the content of the Albizia julibrissin extract is less than 0.00001 wt%, sufficient effect is not exhibited, and even if it is added at 5.0 wt% or more, enhancement of effect cannot be expected.

[0032] The composition containing the Albizia julibrissin extract of the present invention contains an Albizia julibrissin extract as an active ingredient. In the present invention, the effect of suppressing the disturbance of the expression rhythm of clock genes caused by oxidative stress or the like includes the effect on clock genes of physiologically active substances such as polyphenol compounds and terpenoid compounds contained in the Albizia julibrissin extract. It also includes the effect of suppressing the disturbance of the expression rhythm of clock genes in peripheral cells by these effects. Furthermore, in the present invention, the effect of suppressing the expression rhythm of clock genes can be evaluated by adding the composition of the present invention to cultured cells and quantifying the gene expression level in the cells by real-time PCR.

[0033] In the present invention, the clock gene may be any gene that has the function of controlling circadian rhythm (body clock), and is not particularly limited, but examples thereof include Period genes (Per1, Per2, Per3, etc.), Bmal genes (Arntl, Mop3, Tic, Jap3, Pasd3, genes also called bHLHe5, Bmal1, Bmal2, etc.), Cryptochrome genes (Cry1, Cry2, etc.), Clock genes, Ror genes, Rev-erb genes, E4BP genes, GSK3β genes, CK1 genes, and Dec genes, and in the present invention, the Bmal1 gene is preferred. It is also considered that the effect of the clock gene extends to clock-controlled genes whose expression is controlled by the clock gene. It has been reported that 43% of protein-coding genes are controlled by clock genes at some point in the body (PNAS, Vol. 111, No. 45, p16219-16224; 2014), and it is expected that this compound will also have an effect on these clock-controlled genes.

[0034] In the present invention, clock gene expression refers not only to expression at the transcription level (expression as mRNA) but also to expression at the translation level (expression as protein). Furthermore, "changes in clock gene expression" include not only increases or decreases in the expression level of clock genes, but also changes in the expression rhythm of clock genes (at least one of amplitude, phase, and period length).

[0035] "Amplitude" is the width from peak to trough in a rhythm, that is, the magnitude of oscillation. In the expression rhythm of clock genes, it indicates the width of the expression level of the clock genes from peak to peak or from trough to trough. It is known that the amplitude of clock genes is weakened by a decrease in cell activity, aging, stress, etc.

[0036] "Phase" refers to a specific position in one cycle (e.g., peak to peak, trough to trough). "Cycle length" refers to the length of one cycle. As shown in the Examples below, the Albizia julibrissin bark extract can suppress phase retardation and cycle length extension caused by oxidative stress, and maintain normal rhythmic expression of clock genes.

[0037] In this way, the composition containing the extract of the Silkworm of the present invention can suppress abnormalities in the expression rhythm of clock genes and the circadian rhythm controlled thereby, and can improve or prevent symptoms caused by such abnormalities.

[0038] In the present invention, oxidative stress and the like include, for example, oxidative stress caused by hydrogen peroxide, and oxidative stress caused within cells by ultraviolet rays (UV-A, UV-B, UV-C), and the like.

[0039] The composition containing the extract of the present invention may contain other raw materials in addition to the extract of the Albizia julibrissin, provided that the effect of the present invention is not impaired. Examples of such raw materials include water, excipients, antioxidants, preservatives, humectants, thickeners, buffers, adsorbents, solvents, emulsifiers, stabilizers, surfactants, lubricants, water-soluble polymers, sweeteners, flavorings, acidulants, and alcohols. The antioxidant of the present invention may also contain other active ingredients in a range that does not impair the effect of the present invention. Specific examples of active ingredients include antioxidants, anti-aging ingredients, anti-inflammatory ingredients, whitening ingredients, cell activation ingredients, vitamins, blood circulation promoting ingredients, moisturizing ingredients, ingredients that have the effect of preventing and / or repairing DNA damage, anti-glycation ingredients, peptides or derivatives thereof, amino acids or derivatives thereof, hydroquinone glycosides and esters thereof, and the like. In this case, the proportion of the Albizia julibrissin extract in the composition containing the Albizia julibrissin extract is preferably from 0.001 to 10 mass %, more preferably from 0.01 to 5 mass %, and particularly preferably from 0.1 to 2 mass %.

[0040] The Albizia julibrissin bark extract obtained as above can be used as an anti-inflammatory agent as it is, but it is preferable to use it by blending it into an external composition, especially an external composition for skin. In this case, the amount to be blended into the external composition is determined depending on the degree of absorption, the degree of action, the product form, the frequency of use, etc., and is not particularly limited, but it is desirable to make the concentration range of 0.00001 to 5.0 wt% by dry weight, preferably 0.00005 to 3.0 wt%, and particularly preferably 0.0001 to 1.0 wt%. If the content of the Albizia julibrissin bark extract is less than 0.00001 wt%, the effect is not sufficient, and even if it is added at 5.0 wt% or more, the effect cannot be expected to be enhanced.

[0041] The skin external composition containing the Albizia julibrissin bark extract of the present invention contains the Albizia julibrissin bark extract as an active ingredient. The anti-inflammatory effect of the Albizia julibrissin bark extract in the present invention includes the action of physiologically active substances such as polyphenol compounds and terpenoid compounds contained in the Albizia julibrissin bark extract. More specifically, it is believed that the physiologically active substances suppress skin erythema induced by ultraviolet rays and the like. This erythema suppression effect can be evaluated by measuring the skin color after ultraviolet irradiation.

[0042] The Albizia julibrissin bark extract obtained as above can be used as an anti-glycation agent as it is, but it is preferable to use it by blending it into an external composition, especially an external composition for skin. In this case, the amount to be blended into the external composition is determined depending on the degree of absorption, the degree of action, the product form, the frequency of use, etc., and is not particularly limited, but it is desirable to make the concentration range of 0.00001 to 5.0 wt% by dry weight, preferably 0.00005 to 3.0 wt%, and particularly preferably 0.0001 to 1.0 wt%. If the content of the Albizia julibrissin bark extract is less than 0.00001 wt%, the effect is not sufficient, and even if it is added at 5.0 wt% or more, the effect cannot be expected to be enhanced.

[0043] The skin external composition containing the Albizia julibrissin bark extract of the present invention contains the Albizia julibrissin bark extract as an active ingredient. The anti-glycation effect of the Albizia julibrissin bark extract in the present invention includes the action of physiologically active substances such as polyphenol compounds and terpenoid compounds contained in the Albizia julibrissin bark extract. More specifically, it is considered that the physiologically active substances inhibit the reaction between protein and sugar. This anti-glycation effect can be evaluated, for example, by quantifying carbonyl groups as an indicator of glycation products generated by the reaction between protein and sugar.

[0044] In addition to the essential components of the Albizia julibrissin extract, the composition containing the Albizia julibrissin extract of the present invention may contain, as necessary, components and additives used in cosmetics, quasi-drugs, food and beverages, pharmaceuticals, etc., within the scope that does not impair the effects of the present invention.

[0045] Examples of fats and oils include avocado oil, almond oil, fennel oil, perilla oil, olive oil, orange oil, orange raffaella oil, sesame oil, cacao butter, chamomile oil, carrot oil, cucumber oil, beef tallow, beef tallow fatty acids, kukui nut oil, safflower oil, soybean oil, camellia oil, corn oil, rapeseed oil, persic oil, castor oil, cottonseed oil, peanut oil, turtle oil, mink oil, egg yolk oil, cacao butter, palm oil, palm kernel oil, Japan wax, coconut oil, beef tallow, lard, hardened oil, and hardened castor oil.

[0046] Examples of waxes include beeswax, carnauba wax, spermaceti, lanolin, liquid lanolin, reduced lanolin, hard lanolin, candelilla wax, montan wax, and shellac wax.

[0047] Examples of mineral oils include liquid paraffin, vaseline, paraffin, ozokeride, ceresin, microcrystalline wax, polyethylene powder, squalene, squalane, and pristane.

[0048] Examples of fatty acids include natural fatty acids such as lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, 12-hydroxystearic acid, undecylenic acid, tall oil, and lanolin fatty acid, and synthetic fatty acids such as isononanoic acid, caproic acid, 2-ethylbutanoic acid, isopentanoic acid, 2-methylpentanoic acid, 2-ethylhexanoic acid, and isopentanoic acid.

[0049] Examples of alcohols include natural alcohols such as ethanol, isopropanol, lauryl alcohol, cetanol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol, and phytosterol; synthetic alcohols such as 2-hexyldecanol, isostearyl alcohol, and 2-octyldodecanol; and polyhydric alcohols such as ethylene oxide, ethylene glycol, diethylene glycol, triethylene glycol, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, polyethylene glycol, propylene oxide, propylene glycol, polypropylene glycol, 1,3-butylene glycol, glycerin, batyl alcohol, pentaerythritol, sorbitol, mannitol, glucose, and sucrose.

[0050] Examples of esters include isopropyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, oleyl oleate, decyl oleate, octyldodecyl myristate, hexyldecyl dimethyloctanoate, cetyl lactate, myristyl lactate, diethyl phthalate, dibutyl phthalate, lanolin acetate, ethylene glycol monostearate, propylene glycol monostearate, and propylene glycol dioleate.

[0051] Examples of the metallic soap include aluminum stearate, magnesium stearate, zinc stearate, calcium stearate, zinc palmitate, magnesium myristate, zinc laurate, and zinc undecylenate.

[0052] Examples of gums and water-soluble polymer compounds include gum arabic, gum benzoin, gum dammar, guaiac butter, Irish moss, gum karaya, gum tragacanth, carob gum, quince seed, agar, casein, dextrin, gelatin, pectin, starch, carrageenan, carboxyalkyl chitin, chitosan, hydroxyalkyl chitin, low molecular weight chitosan, chitosan salts, sulfated chitin, phosphorylated chitin, alginic acid and its salts, hyaluronic acid and its salts, chondroitin sulfate, heparin, ethyl cellulose, methyl cellulose, carboxymethyl cellulose, carboxyethyl cellulose, sodium carboxyethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, nitrocellulose, crystalline cellulose, polyvinyl alcohol, polyvinyl methyl ether, polyvinylpyrrolidone, polyvinyl methacrylate, polyacrylates, polyalkylene oxides such as polyethylene oxide and polypropylene oxide or crosslinked polymers thereof, carboxyvinyl polymers, and polyethyleneimine.

[0053] Surfactants include anionic surfactants (carboxylates, sulfonates, sulfate salts, phosphate salts), cationic surfactants (amine salts, quaternary ammonium salts), amphoteric surfactants (carboxylate type amphoteric surfactants, sulfate ester type amphoteric surfactants, sulfonate type amphoteric surfactants, phosphate ester type amphoteric surfactants), nonionic surfactants (ether type nonionic surfactants, ether ester type nonionic surfactants, ester type nonionic surfactants, block polymer type nonionic surfactants, nitrogen-containing nonionic surfactants), and other surfactants (natural surfactants, derivatives of protein hydrolysates, polymeric surfactants, surfactants containing titanium and silicon, fluorocarbon surfactants), etc.

[0054] Examples of vitamins include vitamin A group retinol, retinal (vitamin A1), dehydroretinal (vitamin A2), carotene, and lycopene (provitamin A); vitamin B group thiamine hydrochloride, thiamine sulfate (vitamin B1), riboflavin (vitamin B2), pyridoxine (vitamin B6), cyanocobalamin (vitamin B12), folic acid, nicotinic acid, pantothenic acid, biotin, choline, and inositol; vitamin C group ascorbic acid and its derivatives; vitamin D group ergocalciferol (vitamin D2), cholecalciferol (vitamin D3), and dihydrotachysterol; vitamin E group tocopherol and its derivatives and ubiquinones; and vitamin K group phytonadione (vitamin K1), menaquinone (vitamin K2), menadione (vitamin K3), and menadiol (vitamin K4).

[0055] Examples of amino acids include valine, leucine, isoleucine, threonine, methionine, phenylalanine, tryptophan, lysine, glycine, alanine, asparagine, glutamine, serine, cysteine, cystine, tyrosine, proline, hydroxyproline, aspartic acid, glutamic acid, hydroxylysine, arginine, ornithine, histidine, and the like, as well as amino acid derivatives thereof such as sulfates, phosphates, nitrates, citrates, or pyrrolidone carboxylic acids.

[0056] Examples of skin whitening agents include ascorbic acid or a derivative thereof, sulfur, placental hydrolysate, ellagic acid or a derivative thereof, kojic acid or a derivative thereof, glucosamine or a derivative thereof, arbutin or a derivative thereof, hydroxycinnamic acid or a derivative thereof, glutathione, arnica extract, Scutellaria root extract, Sophora japonica extract, Saiko extract, Safflower extract, Ganoderma lucidum mycelium culture or an extract thereof, Tilia extract, peach leaf extract, Ajing fruit extract, Sophora root extract, Jew's wort extract, Angelica acutiloba extract, Coix seed extract, persimmon leaf extract, Rhubarb extract, Peony extract, Hamamelis extract, Horse chestnut extract, Hypericum perforatum extract, and oil-soluble licorice extract.

[0057] Examples of moisturizing agents include hyaluronic acid, polyglutamic acid, serine, glycine, threonine, alanine, collagen, hydrolyzed collagen, hydronectin, fibronectin, keratin, elastin, royal jelly, chondroitin sulfate heparin, glycerophospholipid, glyceroglycolipid, sphingophospholipid, sphingoglycolipid, linoleic acid or esters thereof, eicosapentaenoic acid or esters thereof, pectin, bifidobacterium fermentation product, lactic acid fermentation product, yeast extract, ganoderma lucidum mycelium culture or extract thereof, wheat germ oil, avocado oil, rice germ oil, jojoba oil, soybean phospholipid, γ-oryzanol, bilberry extract, coix seed extract, rehmannia root extract, taiso extract, kaiso extract, aloe arborescens extract, burdock extract, rose wax extract, arnica extract, wheat bran, and the like.

[0058] Hair growth agents include pentadecanoic acid glyceride, coleus extract, gentian extract, matsukasa extract, royal jelly extract, kumazasa extract, t-flavanone, 6-benzylaminopurine, Swertia japonica extract, carpronium chloride, minoxidil, finasteride, adenosine, nicotinamide, mulberry root extract, rehmannia extract, and 5-aminolevulinic acid.

[0059] Examples of extracts and essences from animals, plants, and herbal medicines include Acacia oleracea, Angelica gracilis, Acerola, Altea, Arnica, Avocado, Amacha, Aloe, Aloe vera, Nettle, Ginkgo biloba, Fennel, Turmeric, Asian sardine, Plum, Quercus salicina, Uva-ursi, Rosa multiflora, Astragalus chinensis, Astragalus membranaceus, Scutellaria baicalensis, Cherry bark, Phellodendron amplexicaule, Coptis japonica, Panax ginseng, Hypericum perforatum, and Origanum officinalis. Watercress, Nativity, Orange, Polygala tenuifolia, Prunella vulgaris, Houttuynia cordata, Sophora japonica, Artemisia anguicida, Zedoaria rhizome, Pueraria lobata, Valerian root, Chamomile, Melon root, Artemisia capillaris, Licorice, Coltsfoot, Rubus serrata, Kiwi fruit, Platycodon grandiflorum, Chrysanthemum flower, Catalpa fruit, Citrus fruit, Tachibana peel, Cucumber, Aralia udo or Angelica aralia, Apricot kernel, Kudzu, Goji bark, Goji berries, Goji leaves, Sophora flavonoides, Camphor tree, Kumazasa, Grapefruit fruit, Cinnamon bark, Cinnamon bark, Cassia jasmine, Round morning glory or Japanese morning glory, Safflower, Comfrey, Copaiba, Gardenia, Gentian, Magnolia obovata, Achyranthes jasmine, Euonymus chinensis, Burdock, Schisandra chinensis, Rice, Rice bran, Wheat, Bupleurum fructus, Saffron, Soapwort, Crataegus crenata, Japanese pepper Pepper), Salvia, Sanshichi ginseng, Shiitake mushroom, Rehmannia rhizome, Shikunshi, Purple root, Perilla frutescens, Chinese peony, Plantain, Ginger, Calamus, Ligusticum japonica, Spiraea gracilis, Japanese white birch, Honeysuckle, Ivy, Yarrow, Elderberry, Adzuki bean, Elderberry, Mallow, Cnidium officinalis, Swertia japonica, Mulberry bark,Mulberry leaves), jujube, soybean, taranoki, chikusetsuninjin, anemone, sedge, houttuynia cordata, cordyceps sinensis, chili pepper, ground cherries, thyme, green tea, black tea, cloves, unshu mikan, camellia, centella, chili pepper, touki, touki, tokinsen Moss, bitter orange (orange skin), serroko (elm), corn (southern elm), eucommia ulmoides (Eucommia ulmoides, leaves), tomato, nandina (nanten fruit), garlic, barley (malt), hakusan (white moss skin), Ophiopogon japonica (Ophiopogon), parsley, batata (batata), mint, hamamelis, rose, loquat leaf, matsuhodo (Japanese elm), grape or its leaves, loofah, linden tree, peony (peony skin), hops, mica I (Maiko), pine needles, horse chestnut, rosehip, soapberry, melissa, melilot, Japanese quince, bean sprouts, peach kernel, peach leaf, Chinese bean sprout, areca nut, amplexicaule, centaurea, saxifrage, bayberry, alder bark, alder, coix seed, mugwort, Japanese mugwort, lavender, apple fruit, Ganoderma lucidum, lemon Examples of such extracts include fruit, Forsythia fruit, Astragalus sieboldii, Geranium herb, Scolytemaria root, rooster comb, bovine and human placenta extracts, porcine and bovine stomach, duodenum, or intestine extracts or hydrolyzates thereof, water-soluble collagen, water-soluble collagen derivatives, collagen hydrolyzates, elastin, elastin hydrolyzates, water-soluble elastin derivatives, silk protein, silk protein hydrolyzates, and bovine blood cell protein hydrolyzates.

[0060] Examples of microbial culture metabolites include yeast extract, zinc-containing yeast extract, germanium-containing yeast extract, selenium-containing yeast extract, magnesium-containing yeast extract, rice fermentation extract, Euglena extract, and lactic acid fermentation products of skim milk powder.

[0061] Examples of α-hydroxy acids include glycolic acid, citric acid, malic acid, tartaric acid, and lactic acid.

[0062] Examples of inorganic pigments include silicic anhydride, magnesium silicate, talc, kaolin, bentonite, mica, titanium mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, calcium carbonate, magnesium carbonate, yellow iron oxide, red iron oxide, black iron oxide, ultramarine, chromium oxide, chromium hydroxide, carbon black, and calamine.

[0063] Examples of ultraviolet absorbers include p-aminobenzoic acid derivatives, salicylic acid derivatives, anthranilic acid derivatives, coumarin derivatives, amino acid compounds, benzotriazole derivatives, tetrazole derivatives, imidazoline derivatives, pyrimidine derivatives, dioxane derivatives, camphor derivatives, furan derivatives, pyrone derivatives, nucleic acid derivatives, allantoin derivatives, nicotinic acid derivatives, vitamin B6 derivatives, oxybenzone, benzophenone, guaiazulene, shikonin, baicalin, baicalein, berberine, and the like.

[0064] Examples of astringents include lactic acid, tartaric acid, succinic acid, citric acid, allantoin, zinc chloride, zinc sulfate, zinc oxide, calamine, zinc p-phenolsulfonate, potassium aluminum sulfate, resorcin, ferric chloride, and tannic acid.

[0065] Antioxidants include ascorbic acid and its salts, stearic acid esters, tocopherol and its ester derivatives, nordihydroguaiaretic acid, butylhydroxytoluene (BHT), butylhydroxyanisole (BHA), parahydroxyanisole, propyl gallate, sesamol, sesamolin, gossypol, and the like.

[0066] Anti-inflammatory agents include ichthammol, indomethacin, kaolin, salicylic acid, sodium salicylate, methyl salicylate, acetylsalicylic acid, diphenhydramine hydrochloride, d- or dl-camphor, hydrocortisone, guaiazulene, chamazulene, chlorpheniramine maleate, glycyrrhizic acid and its salts, glycyrrhetinic acid and its salts, and the like.

[0067] Examples of germicides and disinfectants include acrinol, sulfur, benzalkonium chloride, benzethonium chloride, methylrosaniline chloride, cresol, calcium gluconate, chlorhexidine gluconate, sulfamine, mercurochrome, lactoferrin, or its hydrolysates.

[0068] Examples of hair care products include selenium disulfide, alkylisoquinolinium bromide solution, zinc pyrithione, biphenamine, thianthol, castor tincture, ginger tincture, capsicum tincture, quinine hydrochloride, strong aqueous ammonia, potassium bromate, sodium bromate, and thioglycolic acid.

[0069] Fragrances include natural animal fragrances such as musk, civet, castoreum, and ambergris, anise essential oil, angelica essential oil, ylang ylang essential oil, iris essential oil, fennel essential oil, orange essential oil, cananga essential oil, caraway essential oil, cardamom essential oil, guaiac wood essential oil, cumin essential oil, black letter essential oil, cassia bark essential oil, cinnamon essential oil, geranium essential oil, copaiba balsam essential oil, coriander essential oil, shiso essential oil, cedarwood essential oil, citronella essential oil, jasmine essential oil, ginger grass essential oil, cedar essential oil, spearmint essential oil, and western Examples of plant-based fragrances include peppermint essential oil, anise essential oil, tuberose essential oil, clove essential oil, orange flower essential oil, wintergreen essential oil, tolu balsam essential oil, butterbur essential oil, rose essential oil, palmarosa essential oil, cypress essential oil, Japanese cedar essential oil, sandalwood essential oil, petitgrain essential oil, bay essential oil, vetiver essential oil, bergamot essential oil, Peru balsam essential oil, bois de rose essential oil, camphor essential oil, mandarin essential oil, eucalyptus essential oil, lime essential oil, lavender essential oil, linaloe essential oil, lemongrass essential oil, lemon essential oil, rosemary essential oil, and Japanese peppermint essential oil, as well as other synthetic fragrances.

[0070] Examples of pigments and coloring agents include red cabbage pigment, red rice pigment, madder pigment, annatto pigment, squid ink pigment, turmeric pigment, sophora japonica pigment, krill pigment, persimmon pigment, caramel, gold, silver, gardenia pigment, corn pigment, onion pigment, tamarind pigment, spirulina pigment, whole buckwheat pigment, cherry pigment, seaweed pigment, hibiscus pigment, grape juice pigment, marigold pigment, purple sweet potato pigment, purple yam pigment, lac pigment, and rutin.

[0071] Examples of sweeteners include sugar, sweet tea, fructose, arabinose, galactose, xylose, mannose, maltose, honey, glucose, miraculin, and monellin.

[0072] Nutritional supplements include calcined shell calcium, cyanocolabamin, yeast, wheat germ, soybean germ, egg yolk powder, hemicellulose, and heme iron.

[0073] Other examples include hormones, sequestering agents, pH adjusters, chelating agents, preservatives and antimicrobial agents, cooling agents, stabilizers, emulsifiers, animal and plant proteins and their decomposition products, animal and plant polysaccharides and their decomposition products, animal and plant glycoproteins and their decomposition products, blood flow promoters, anti-inflammatory and anti-allergic agents, cell activators, keratolytic agents, wound healing agents, foaming agents, thickening agents, oral agents, deodorizing and deodorant agents, bittering agents, seasonings, enzymes, etc.

[0074] The dosage form of the present invention is arbitrary, and can be used by blending it with quasi-drugs such as ampoules, capsules, powders, granules, pills, tablets, solids, liquids, gels, foams, emulsions, creams, ointments, sheets, and mousses, cosmetics for the skin and hair, bath preparations, foods, beverages, and medicines.

[0075] Specific examples of cosmetics and quasi-drugs include basic cosmetics such as internal and external medicinal preparations, lotions, milky lotions, creams, ointments, lotions, oils and packs, face washes and skin cleansers, hair cosmetics such as shampoos, rinses, hair treatments, hair creams, pomades, hair sprays, hair styling products, perm agents, hair tonics, hair dyes, hair growth and care products, make-up cosmetics such as foundations, face powders, powders, lipsticks, blushers, eye shadows, eyeliners, mascara, eyebrow inks and eyelashes, finishing cosmetics such as nail polish, perfumes, bath products, and others.

[0076] Examples of food and beverages include beverages such as soft drinks, carbonated drinks, nutritional drinks, fruit drinks, and lactic acid drinks; cold desserts such as ice cream, ice sherbet, and shaved ice; noodles such as soba, udon, vermicelli, gyoza wrappers, shumai wrappers, Chinese noodles, and instant noodles; sweets such as candy, candy, gum, chocolate, tablet candy, snacks, biscuits, jelly, jam, cream, baked goods, and bread; seafood such as crab, salmon, clams, tuna, sardines, shrimp, bonito, mackerel, whale, oysters, pacific saury, squid, ark shells, scallops, abalone, sea urchin, salmon roe, and tokobushi; and water products such as kamaboko, ham, and sausage. These include processed dairy and livestock foods, dairy products such as milk powder, processed milk, fermented milk, salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, dressings, and other oils and fats and processed oil foods, seasonings such as sauces and dressings, retort pouch foods such as curry, stew, oyakodon, porridge, zosui, Chinese rice bowls, katsudon, tendon, unadon, hayashi rice, oden, mabo dolph, beef bowls, meat sauce, egg soup, omelet rice, gyoza, shumai, hamburger steaks, and meatballs, various forms of health and nutritional supplements, health functional foods, tablets, capsules, drinks, troches, etc.

[0077] The composition containing an extract of Albizia julibrissin of the present invention is preferably applied to humans, but can also be applied to animals other than humans as long as the respective action and effect can be expected.

[0078] The form of the cosmetic of the present invention is not particularly limited, but examples include facial cleansers, cleansers, lotions (e.g., whitening lotions), creams (e.g., vanishing cream, cold cream), milky lotions, gels, beauty serums, packs (e.g., jelly-like peel-off type, paste-like wipe-off type, powder-like wash-off type), face masks, cleansing, foundations, lipsticks, lip balms, lip glosses, lip liners, blushers, makeup bases, shaving lotions, sunscreens, after-sun lotions, deodorant lotions, body lotions (including hand care lotions and foot care lotions), body oils, and the like.

[0079] The present invention will be described in more detail below based on examples, although the present invention is not particularly limited by the examples. EXAMPLES

[0080] The water extract of Albizia julibrissin bark used in the following examples was prepared by the method disclosed in JP 2018-58783 A.

[0081] Example 1: Evaluation of the inhibitory or preventive effect on disruption of the expression rhythm of target clock genes in normal human neonatal epidermal keratinocytes The above-mentioned Albizia julibrissin extract was used as a test sample, and the expression rhythm of clock genes was evaluated by the following test method.

[0082] Human normal neonatal epidermal keratinocytes (NHEK) were seeded on a collagen-coated T75 flask using Collagen Coating Solution (Toyobo), and cultured at 37°C with 10% CO2 after adding normal human epidermal keratinocyte growth medium (HuMedia-KG2). Culture was continued until the cells reached 80% or more confluence, with appropriate medium changes. After removing the medium and rinsing with PBS, the cells were harvested by trypsinization. 1 × 10 4 The cells were seeded at 10 cells / well and cultured at 37°C, 10% CO2 for 1 day.

[0083] The above-mentioned cultured cells were supplemented with a medium containing dexamethasone dissolved therein to a final concentration of 100 nM, incubated for 1 hour, and stimulated to synchronize. The medium was then removed, replaced with a medium containing hydrogen peroxide and the test sample at the designated concentrations, and incubated for 24 hours at 37°C and 10% CO2. In the hydrogen peroxide-free / silm extract-free group, hydrogen peroxide was 0 mM / silm extract 0 mg / mL, in the hydrogen peroxide-added / silm extract-free group, hydrogen peroxide was 1.5 mM / silm extract 0 mg / mL, and in the hydrogen peroxide-added / silm extract-added group, hydrogen peroxide was 1.5 mM / silm extract 50 μg / mL. The cells were harvested at regular intervals during the 24-hour incubation, the medium was removed, and the cells were frozen and stored at -80°C. After all levels of cells had been collected and frozen, cDNA was prepared from the cells using SuperPrep (registered trademark) II Cell Lysis & RT Kit for qPCR (manufactured by Toyobo).

[0084] Based on the obtained cDNA, the mRNA gene was amplified by qRT-PCR using THUNDERBIRD SYBR (registered trademark) qPCR Mix (manufactured by Toyobo Co., Ltd.). The reaction solution composition was in accordance with the attached instructions. However, the reaction solution volume was 20 μl / well, and the cDNA volume was 3 μl. The reaction cycle conditions were 95°C, 1 minute → (95°C, 15 seconds → 60°C, 45 seconds) × 40 → 95°C, 15 seconds → 60°C, 1 minute → 95°C, 15 seconds. The primers used were the base sequences shown in SEQ ID NO: 1 to 2. The equipment used was the 7500 Fast Real-Time PCR System (Applied Biosystems). The expression level of the target gene was standardized using the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene as an internal standard (SEQ ID NO: 3 to 4). The normalized expression level of each target gene was divided by the expression level of the control gene, which was collected immediately after the completion of synchronous stimulation without adding hydrogen peroxide or test sample, to calculate a relative value.

[0085] The results are shown in Figures 1 to 3. In the hydrogen peroxide-free / silvus julibrissin extract-free group in Figure 1, the Bmal1 expression cycle was approximately 24 hours, with a normal expression rhythm peaking at 10 to 11 hours. In the hydrogen peroxide-added / silvus julibrissin extract-free group in Figure 2, the Bmal1 expression cycle was more than 24 hours, with an irregular or abnormal expression rhythm of 12 to 18 hours with no clear peak. In the hydrogen peroxide-added / silvus julibrissin extract-added group in Figure 3, the Bmal1 expression cycle was approximately 24 hours, with a peak at 10 to 12 hours, and was confirmed to be close to the normal expression rhythm.

[0086] The above results demonstrated that the extract of Albizia julibrissin has the effect of suppressing or preventing the irregular or abnormal rhythm of clock gene expression caused by oxidative stress due to hydrogen peroxide in epidermal cells.

[0087] Example 2: Formulation of topical skin preparations The following are examples of formulations of the composition containing the Albizia julibrissin extract of the present invention as topical skin preparations. All of these preparations are expected to have the effect of suppressing or preventing abnormalities in the expression rhythm of clock genes caused by the Albizia julibrissin extract.

[0088] Toner A lotion was prepared in a conventional manner according to the following composition. ·Purified water ···89.80g Glycerin: 3.00 g Phenoxyethanol: 0.20 g Butylene glycol 5.00 g Pentylene glycol 1.00 g Silk tree extract: 1.00 g

[0089] Gel A gel was prepared in a conventional manner according to the following composition: Purified water: 88.50g Carbomer 0.30 g Xanthan gum 0.10g Arginine: 0.40 g Glycerin: 5.00 g Phenoxyethanol: 0.20 g Butylene glycol 5.00 g Silk extract: 0.50 g

[0090] cream A cream was prepared in a conventional manner according to the following composition. Purified water: 58.50g Butylene glycol 10.00 g Glycerin: 5.00 g Phenoxyethanol: 0.20 g Ethylhexylglycerin: 0.20g Squalane: 10.00g Olive oil: 10.00g Behenyl alcohol 2.50g Polyglyceryl-10 pentastearate 1.90g Sodium stearoyl lactate 0.60g Cetyl palmitate 1.00g Silk extract: 0.10g

[0091] Example 3: Erythema Inhibition Test The above-mentioned Albizia julibrissin bark extract was used as a test sample, and an erythema inhibition test was carried out according to the following test method.

[0092] subject The subjects were 12 men and women in their 20s and 30s, as shown in Table 1. The average age of the subjects was 33.8 ± 4.9 years (21 to 39 years).

[0093] [Table 1]

[0094] Test sample The water extract (powder) of the albizia julibrissin bark was dissolved in a 30% butylene glycol solution to a concentration of 0.01% by mass, and the solution was used as a test sample.

[0095] Induction of erythema by ultraviolet irradiation The UV irradiation was performed using a Multiport SOLAR UV Simulator Model 601 (SOLAR Light Co. Inc., wavelength range: 290-400 nm, peak wavelength: 356 nm) as a light source, and the irradiation intensity was measured using a multi-function measurement system model PMA2100 (SOLAR Light Co. Inc.). First, 0.64, 0.80, 1.00, 1.25, 1.56, and 1.95 MED of UV was irradiated onto the inner side of the upper arm of each subject (MED determination irradiation). 24 hours after irradiation, the MED of each subject was determined (MED determination). Next, UV equivalent to 1.5 MED was irradiated onto the inner side of the left upper arm of each subject, creating four pigmented areas with a diameter of approximately 1 cm (main irradiation). 24 hours after main irradiation, skin color was measured.

[0096] Use of test samples The test sample was applied to the inside of the upper arm in an appropriate amount twice a day for three days: the day of irradiation for MED evaluation, the day of MED evaluation and main irradiation, and the day of skin color measurement.

[0097] skin color measurement Before and one day after the main irradiation, the skin color was measured in a pre-set analysis range of the UV-irradiated area and the adjacent non-irradiated area using a spectrophotometer (CM2600d; manufactured by Konica Minolta). The a* value was used as an index of erythema. Each area was measured 3 to 7 times, and the average of the three stable measurements was used. The degree of erythema was calculated as Δa* value = a* value (UV-irradiated area) - a* value (non-irradiated area). Furthermore, the Δa* value (24 hours later) - Δa* value (before irradiation) was calculated and compared for the test sample application area and the non-application area to evaluate the degree of erythema suppression.

[0098] The results are shown in Figure 4. When comparing the test sample application area with the non-application area, the change in Δa* value (change from before irradiation to 24 hours after irradiation) was suppressed in the test sample application area. This confirmed that the skin erythema induced by UV irradiation was suppressed.

[0099] Example 4: Formulation of a skin topical composition Examples of formulations of the skin external composition containing the Albizia julibrissin bark extract of the present invention are shown below. All of these preparations are expected to have an erythema suppression effect caused by the Albizia julibrissin bark extract and are effective as anti-inflammatory agents.

[0100] Toner A lotion was prepared in a conventional manner according to the following composition. ·Purified water ···89.80g Glycerin: 3.00 g Phenoxyethanol: 0.20 g Butylene glycol 5.00 g Pentylene glycol 1.00 g Silk tree bark extract: 1.00 g

[0101] Gel A gel was prepared in a conventional manner according to the following composition: Purified water: 88.50g Carbomer 0.30 g Xanthan gum 0.10g Arginine: 0.40 g Glycerin: 5.00 g Phenoxyethanol: 0.20 g Butylene glycol 5.00 g Silk tree bark extract: 0.50 g

[0102] cream A cream was prepared in a conventional manner according to the following composition. Purified water: 58.50g Butylene glycol 10.00 g Glycerin: 5.00 g Phenoxyethanol: 0.20 g Ethylhexylglycerin: 0.20g Squalane: 10.00g Olive oil: 10.00g Behenyl alcohol 2.50g Polyglyceryl-10 pentastearate 1.90g Sodium stearoyl lactate 0.60g Cetyl palmitate 1.00g Silk tree bark extract: 0.10 g

[0103] Example 5: Glycation inhibition test The above-mentioned Albizia julibrissin bark extract was used as a test sample, and a glycation inhibition test was carried out according to the following test method.

[0104] A BSA-glucose mixture was prepared by dissolving 1.6 g of bovine serum albumin (BSA; RIA grade) and 3.0 g of D-glucose in 10 mL of PBS. The solution was filtered through a 0.45 μm filter, and then 10% of a 30% butylene glycol (BG) solution in which 1.0% by mass of the test sample (powder) was dissolved was added to the solution, and the solution was incubated at 45° C. for 10 days. As a control addition group, a level in which 10% of 30% BG solution was added was prepared, since the above sample contained 30% BG. A mixed sample prepared in the same manner was stored at −80° C. as a negative control group. After 10 days, each reaction solution was diluted 10 times with purified water, and 100 μmL was dispensed into a 96-well microplate. Next, NBD-H (4-Hydrazino-7-nitro-2,1,3-benzoxadiazole Hydrazine) was added at a final concentration of 250 μmol / L in the presence of trifluoroacetic acid at a final concentration of 0.025%. The reaction was allowed to proceed for 15 minutes at room temperature in a dark environment, and the fluorescence intensity was measured using a microplate reader (Ex / Em=470 nm / 550 nm). The amount of carbonyl groups in the reaction solution was determined from a calibration curve created at the same time using propionaldehyde.

[0105] The results are shown in Table 2 and Figure 5. A significant decrease in carbonyl groups was confirmed in the test sample-added group compared to the control-added group, confirming that the production of glycation products caused by the reaction of proteins with sugars was suppressed.

[0106] [Table 2]

[0107] Example 6: Formulation of a skin topical composition Examples of formulations of the skin external composition containing the Albizia julibrissin bark extract of the present invention are shown below. All of these preparations are expected to have an anti-glycation effect due to the Albizia julibrissin bark extract, and are effective as anti-glycation agents.

[0108] Toner A lotion was prepared in a conventional manner according to the following composition. ·Purified water ···89.80g Glycerin: 3.00 g Phenoxyethanol: 0.20 g Butylene glycol 5.00 g Pentylene glycol 1.00 g Silk tree bark extract: 1.00 g

[0109] Gel A gel was prepared in a conventional manner according to the following composition: Purified water: 88.50g Carbomer 0.30 g Xanthan gum 0.10g Arginine: 0.40 g Glycerin: 5.00 g Phenoxyethanol: 0.20 g Butylene glycol 5.00 g Silk tree bark extract: 0.50 g

[0110] cream A cream was prepared in a conventional manner according to the following composition. Purified water: 58.50g Butylene glycol 10.00 g Glycerin: 5.00 g Phenoxyethanol: 0.20 g Ethylhexylglycerin: 0.20g Squalane: 10.00g Olive oil: 10.00g Behenyl alcohol 2.50g Polyglyceryl-10 pentastearate 1.90g Sodium stearoyl lactate 0.60g Cetyl palmitate 1.00g Silk tree bark extract: 0.10 g [Industrial Applicability]

[0111] By using the composition containing the Albizia julibrissin extract of the present invention, it is possible to suppress or prevent abnormalities in the expression rhythm of clock genes in skin cells that show irregular or abnormal expression rhythms of clock genes compared to normal expression rhythms of clock genes, and it is expected to prevent various diseases and symptoms caused by disturbances in the expression rhythm of clock genes. In addition, by using an anti-inflammatory agent or a skin topical composition containing the Albizia julibrissin bark extract of the present invention, it is possible to suppress erythema induced by ultraviolet rays, etc., and it is expected to be used as an anti-inflammatory agent. Furthermore, by using an anti-glycation agent and a skin topical composition containing the Albizia julibrissin bark extract of the present invention, it is possible to effectively suppress the production of glycation products caused by the reaction of proteins and sugars, and it is expected to be used as an anti-glycation agent. Both of them can be applied to skin topical agents, particularly as cosmetics or quasi-drugs.

Claims

1. A cosmetic composition containing an extract of Albizia julibrissin, which is used on any skin cell selected from the group consisting of human epidermal keratinocytes, human dermal fibroblasts, human epidermal melanocytes, human dermal microvascular endothelial cells, human dermal microlymphatic endothelial cells, and human hair papilla cells, which exhibits an irregular or abnormal clock gene expression rhythm compared to the normal clock gene expression rhythm, for suppressing or preventing abnormality in the clock gene expression rhythm.

2. The composition containing an extract of Albizia julibrissin for use in cosmetics according to claim 1, wherein the irregular or abnormal expression rhythm of clock genes is caused by any factor selected from the group consisting of oxidative stress, ultraviolet rays, external stimuli due to drugs, physiological stress, and internal factors due to aging.

3. A composition containing an extract of the silk tree for use in cosmetics as described in claim 2, wherein the irregular or abnormal expression rhythm of clock genes is caused by oxidative stress.

4. The composition containing an extract of Albizia julibrissin for use as a cosmetic preparation according to any one of claims 1 to 3, wherein the irregular or abnormal expression rhythm of the clock gene is a phase delay.

5. The composition containing an extract of Albizia julibrissin for use as a cosmetic preparation according to claim 1 , wherein the clock gene is a gene that exhibits circadian periodicity in its expression rhythm.

6. The composition for use as a cosmetic preparation according to any one of claims 1 to 5, wherein the clock gene is at least one gene selected from the group consisting of a Per gene, a Bmal gene, a Clock gene, a Cry gene, a Tim gene, an Npas gene, an Nr1d1 gene, and a Dbp gene.

7. The cosmetic composition containing an extract of Albizia julibrissin according to claim 6, wherein the clock gene is the Bmal1 gene.

8. The composition for cosmetic use containing an extract of Albizia julibrissin according to any one of claims 1 to 7, wherein the extract of Albizia julibrissin is an extract of Albizia julibrissin bark.

9. The composition for cosmetic use containing an extract of Albizia julibrissin according to any one of claims 1 to 8, wherein the extract of Albizia julibrissin is present in an amount of 0.001 to 10% by mass of the total composition to be applied to the skin cells.

10. The composition according to claim 9, wherein the Albizia julibrissin extract is 0.005 to 1% by mass of the total composition to be applied to the skin cells.

11. A cosmetic comprising the composition containing an extract of Albizia julibrissin according to any one of claims 1 to 10.

12. The cosmetic preparation according to claim 11, which is in any form selected from the group consisting of a lotion, a gel, and a cream.

13. A skin external preparation comprising a composition containing an extract of Albizia julibrissin according to any one of claims 1 to 10.

14. The skin external preparation according to claim 13, wherein the extract of Albizia julibrissin is contained in an amount of 0.00001 to 5.0% by dry weight.

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