Promoter for lipid synthesis in sebaceous gland cells

Abstract

The present invention provides a novel promoter for lipid synthesis in sebaceous gland cells. By using the novel promoter for lipid synthesis in sebaceous gland cells, a skin barrier damage repairing agent and cosmetics with a skin barrier damage repairing function can be prepared, so that cosmetics for repairing skin barrier damage of at least one type selected from skin dryness, premature skin aging, skin aging, skin pruritus, and senile sebaceous deficiency eczema caused by hypofunction of sebaceous glands and / or reduced sebum secretion can be developed.

Description

Promoter of lipid synthesis by sebocytes TECHNICAL FIELD

[0001] The present application relates to a new promoter of lipid synthesis by sebocytes, the use of the promoter of lipid synthesis by sebocytes in the preparation of a skin barrier damage repair agent, and the use of the promoter of lipid synthesis by sebocytes in the preparation of a cosmetic for promoting skin barrier damage repair. BACKGROUND

[0002] Sebaceous glands are important skin appendages, whose basic physiological function is to synthesize and secrete sebum to the skin surface to moisturize the skin and hair. The anatomical location of sebaceous glands is often closely associated with hair follicles, forming a pilosebaceous unit. There are about two million sebaceous glands in human skin, with a higher density on the face, about 400-900 per square centimeter. Sebaceous glands are composed of two parts, the duct and the gland. The duct opens in the infundibulum of the hair follicle, and the gland is composed of multiple layers of sebocytes, which realize cell renewal and sebum synthesis and secretion through continuous proliferation and differentiation. Undifferentiated sebocytes are distributed in the peripheral zone of the gland and have the ability to proliferate. The newly generated daughter cells migrate to the center and gradually differentiate, lose the ability to proliferate, and begin to synthesize lipids. Intracellular lipids gradually increase and are stored in lipid droplets. In the central necrotic zone, mature sebocytes become large in size and finally differentiate into mature sebocytes, which release lipids in a holocrine secretion manner, i.e., cell autolysis, and secrete them into the infundibulum of the hair follicle, and then to the body surface. The sebum secreted by sebaceous glands contains 56% triglycerides and their hydrolysis products (diglycerides and free fatty acids), 26% wax esters, 15% squalene, 3% cholesterol esters, and 1.5% cholesterol. Sebum and sweat secreted by sweat glands emulsify on the skin surface to form a hydro-lipid film, which covers the surface of the stratum corneum and is the outermost line of defense of the skin.

[0003] This continuous differentiation process and lipid synthesis of sebocytes is tightly regulated by a variety of paracrine, endocrine hormones and neural mediators. Androgen is the first endocrine hormone recognized to regulate sebaceous secretion. Androgen promotes the proliferation, differentiation of sebocytes and the synthesis and secretion of sebum by activating androgen receptors in sebocytes. Further studies have found that sebaceous glands are not only regulated by androgens, but also by almost all endocrine hormones, including various hormones secreted by the endocrine system such as hypothalamus, pituitary, adrenal gland, thyroid gland, pancreas, etc., such as Corticotrophin Releasing Hormone (CRH), Adrenocorticotropic Hormone (ACTH), Melanocyte-stimulating hormone (MSH), glucocorticoids, prolactin, thyroid hormone, growth hormone, insulin and insulin-like growth factor (IGF-1), etc. More recent studies have found that endocannabinoids play a key regulatory role in sebaceous gland function. Human sebocytes can produce endocannabinoids such as N-arachidonoylethanolamide (AEA) and 2-arachidonoylglycerol (2-AG), and these locally produced endocannabinoids stimulate sebocyte differentiation and lipid synthesis by acting on cannabinoid receptor 2 (CB2) on sebocytes and activating the ERK1 / 2 MAPK signaling pathway mediated by CB2.

[0004] Hypersecretion of sebum not only leads to greasy skin and enlarged pores, but also is the main cause of skin pathological conditions such as acne and seborrheic dermatitis. Therefore, "oil control" (i.e., inhibiting sebaceous gland activity and reducing sebum secretion) has always been the main aspect of sebaceous gland regulation. Patent document 1 (CN116747213A published on September 15, 2023) discloses the application of emodin in inhibiting the synthesis and secretion of sebaceous glands.

[0005] With the deepening of research, people gradually realized that sebaceous glands actually have many important functions, such as maintaining skin barrier, regulating inflammatory response, delaying aging, etc. Sebum, as an important component of skin barrier, is very important in improving the water-retaining capacity and antioxidant capacity of epidermis. The protective water-fat film formed by the emulsification of sebum and sweat on the skin surface not only has the effect of moisturizing the skin and making the skin smooth and tender, but also effectively prevents trans-epidermal water loss (TEWL) and locks water. Sebum also participates in the organization of the three-dimensional structure of skin surface lipids, which helps to maintain the integrity of the skin barrier. Glycerol, which is generated by the hydrolysis of triglyceride, a component of sebum, is a key component of water in the stratum corneum. Sebum is rich in antioxidants such as squalene, vitamin E, coenzyme Q10, etc., which can reduce oxidative damage caused by ultraviolet radiation. Lipase produced by microorganisms on the skin surface decomposes triglycerides in sebum into free fatty acids, which ensures the weakly acidic environment on the skin surface, not only inhibits the colonization of pathogenic bacteria and maintains the balance of skin microecology, but also regulates the activity of proteases such as desquamating enzymes and lipid processing enzymes, which is beneficial to the maintenance and repair of the skin barrier. The lipids secreted by sebaceous glands, such as lauric acid, palmitic acid, sapienic acid, linoleic acid, and short-chain fatty acids generated by propionic acid bacteria metabolism, have direct antibacterial activity and pro-inflammatory or anti-inflammatory activity. It can be seen that sebum and cutaneous microorganisms interact with each other to maintain skin homeostasis.

[0006] Dysfunction of sebaceous glands can cause abnormal changes in the amount and composition of sebum secretion, and is involved in the pathological development of various skin diseases. More and more research evidence shows that atopic dermatitis (AD) is closely related to hyposeborrhoea. Children with AD often have hypoplastic sebaceous glands and smaller volume, and the disease gradually subsides in adolescence, which is consistent with the increasing trend of sebum secretion in adolescence. In adulthood, it can develop into dry skin. Studies have confirmed that the sebaceous glands of AD patients are atrophic, and the amount of sebum secretion is lower than that of healthy people, especially the content of squalene and wax esters in sebum. The low content of sebum in AD patients is often associated with weakened epidermal hydration, so there is a certain relationship between the destruction of the skin barrier function of AD patients and the reduction of sebaceous gland function. Some studies have found that the use of topical cream containing endocannabinoids can stimulate sebum production, improve AD symptoms, and reduce the use of glucocorticoids, so local stimulation of sebum production may be a new method for the treatment of AD. Sebaceous glands are also involved in the regulation of skin aging. In general, the aging rate of oily skin is significantly slower than that of dry skin, which is mainly due to the fact that healthy oily skin secretes more sebum, forms a dense sebum film, locks in moisture, ensures the water needed for normal cell metabolism, maintains skin homeostasis, and blocks external stimuli to enhance the protective ability of the skin. The morphology and activity of sebaceous glands in the elderly will change significantly. With age, sebaceous glands become smaller, function declines, sebum secretion decreases, and sebum film is incomplete, making the skin more susceptible to external stimuli, causing barrier dysfunction, and increasing transdermal water loss. The destruction of the sebum film also changes the weakly acidic environment on the surface of the skin, affecting the activity of various proteases, such as activating serine proteases, causing itching, and in severe cases, leading to the onset of pruritus and asteatotic eczema in the elderly. Pruritus in the elderly is mostly caused by a lack of sebum, often occurring on the limbs and back, accompanied by dry skin and desquamation. In winter and spring, the weather becomes cold and dry, and the elderly often take hot water baths, which can wash away a lot of sebum, leading to increased skin dryness and itching, and repeated scratching can develop into asteatotic eczema. Clinical studies have found that moisturizing creams containing endocannabinoids can more effectively treat asteatotic eczema in the elderly than traditional moisturizing creams. It can be seen that the secretion status of sebum is one of the important factors that determine the health of the skin, and both excessive and insufficient sebum secretion and changes in lipid composition can affect the skin and are closely related to the occurrence and development of various skin diseases. A deeper understanding of the biological function of sebaceous glands and its regulatory mechanism and the discovery of regulators of sebum production will help to develop new methods for the prevention and treatment of sebaceous gland-related diseases.

[0007] Prior art documents

[0008] Patent documents

[0009] Patent Literature 1 CN116747213A published on September 15, 2023 SUMMARY

[0010] PROBLEMS TO BE SOLVED BY THE INVENTION

[0011] An object of the present invention is to find a new promoter of lipid synthesis by sebocyte, and to prepare a new skin barrier damage repair agent using the promoter of lipid synthesis by sebocyte, for repairing skin barrier damage caused by hyposecretion of sebaceous glands and / or reduction in sebaceous glands secretion, and further, an object of the present invention is to provide a cosmetic for promoting skin barrier damage repair, which can repair skin barrier damage caused by hyposecretion of sebaceous glands and / or reduction in sebaceous glands secretion.

[0012] MEANS FOR SOLVING THE PROBLEMS

[0013] The present inventors have found that neryl alcohol can promote lipid synthesis by sebocyte, i.e., have found a new promoter of lipid synthesis by sebocyte, and the neryl alcohol can be prepared into a new promoter of lipid synthesis by sebocyte, and further, can be used for preparing a cosmetic having a skin barrier damage repair function, and thus have completed the present invention.

[0014] That is, the present invention includes the following.

[0015] 1. A promoter of lipid synthesis by sebocyte, characterized by containing neryl alcohol.

[0016] 2. Use of neryl alcohol in the production of a promoter of lipid synthesis by sebocyte.

[0017] 3. Use of neryl alcohol in the production of a skin barrier damage repair agent, the skin barrier damage being skin barrier damage caused by hyposecretion of sebaceous glands and / or reduction in sebaceous glands secretion.

[0018] 4. Use of neryl alcohol in the production of a cosmetic for promoting skin barrier damage repair, the skin barrier damage being skin barrier damage caused by hyposecretion of sebaceous glands and / or reduction in sebaceous glands secretion.

[0019] 5. The use according to the above 3 or 4, the skin barrier damage being at least one of skin dryness, skin premature aging, skin aging, skin pruritus, and senile sebaceous deficiency eczema caused by hyposecretion of sebaceous glands and / or reduction in sebaceous glands secretion.

[0020] EFFECT OF THE INVENTION

[0021] According to the present application, a new sebocyte lipid synthesis promoter can be provided, and using the new sebocyte lipid synthesis promoter, a skin barrier damage repair agent can be prepared, and a cosmetic having a skin barrier damage repair function can be prepared, and thus a cosmetic for repairing at least one of skin barrier damage caused by sebaceous gland hypofunction and / or reduced sebaceous secretion, skin dryness, skin premature aging, skin aging, skin pruritus, and senile sebaceous deficiency eczema can be developed. BRIEF DESCRIPTION OF DRAWINGS

[0022] FIG. 1 is a graph showing the results of a fluorescent enzyme reader analysis of the promotion of lipid synthesis by neral in SZ95 human sebocytes.

[0023] FIG. 2 is a graph showing the results of a flow cytometry analysis of the promotion of lipid synthesis by neral in SZ95 human sebocytes.

[0024] FIG. 3 is a graph showing the results of a confocal microscope analysis of the promotion of lipid droplet accumulation by neral in SZ95 human sebocytes.

[0025] FIG. 4 is a graph showing the effect of neral on the inhibition of proliferation in SZ95 human sebocytes.

[0026] FIG. 5 is a graph showing the effect of neral on the increase in granularity in SZ95 human sebocytes.

[0027] FIG. 6 is a graph showing the effect of neral on the induction of differentiation marker gene expression in SZ95 human sebocytes.

[0028] FIG. 7 is a graph showing the effect of neral on the levels of transcription factors and fatty acid synthesis enzyme proteins that regulate sebum production. DETAILED DESCRIPTION

[0029] The neral, cis-3,7-Dimethyl-2,6-octadienol, described in the present application has a molecular formula of C 10 H 18O, molecular weight 154.25, has a pleasant rose and orange flower aroma, the aroma is relatively mild, with a slight lemon-like fruity aroma. Nerolidol is an isomer of geraniol, its aroma is softer and more elegant than geraniol, relatively light and fresh, with a citrus flavor. Nerolidol is a known monoterpene alcohol compound, which exists in many plant essential oils, such as citrus essential oil and citronella essential oil. Nerolidol was first isolated from neroli oil, which is often used as an aromatic agent in aromatherapy for the treatment of palpitations, rheumatism, anxiety and depression, etc. Nerolidol has been found to have analgesic, anti-inflammatory, antioxidant, anticancer, antiarrhythmic, hepatoprotective, antispasmodic and antibacterial activities. In addition, studies on an oxazolone-induced mouse model of ulcerative colitis found that nerolidol had significant analgesic and anti-inflammatory effects, indicating its potential for treating ulcerative colitis. Nerolidol can also effectively protect the rat liver from liver damage caused by paracetamol through antioxidant and anti-inflammatory effects. In addition, nerolidol has recently been found to activate the olfactory receptor OR2W3 expressed on human airway smooth muscle cells, thereby promoting airway smooth muscle cell and airway smooth muscle tissue relaxation, indicating its potential for treating obstructive lung disease.

[0030] The present inventors have found for the first time that nerolidol has the function of promoting the synthesis of lipids by sebocytes, and therefore nerolidol can be used as a promoter of lipid synthesis by sebocytes.

[0031] The promoter of lipid synthesis by sebocytes described in the present application refers to a promoter having the function of promoting lipid synthesis by sebocytes.

[0032] It is known that hyposecretion of sebaceous glands and / or reduced secretion of sebum can cause skin barrier damage. Therefore, the function of nerolidol found by the present inventors to promote the synthesis of lipids by sebocytes can be used to prepare a new promoter of lipid synthesis by sebocytes, and further used to prepare a skin barrier damage repair agent and a cosmetic having a skin barrier damage repair function.

[0033] The skin barrier damage described in the present application refers to skin barrier damage caused by hyposecretion of sebaceous glands and / or reduced secretion of sebum. Specifically, at least one of the following can be listed: dry skin, premature skin aging, skin aging, skin pruritus, and senile seborrhea.

[0034] The "cosmetic having a skin barrier damage repair function" described in the present application refers to any cosmetic having a skin barrier damage repair function for the skin from the outside, for example, an aqueous solution preparation such as a skin toner, a skin softener, a skin refresher, a skin repairer, and the like; a spray preparation such as a moisturizing spray, an anti-wrinkle spray, a skin repair spray, and the like; an emulsion preparation such as a moisturizing emulsion, an anti-wrinkle emulsion, a skin repair emulsion, a body emulsion, a hand emulsion, and the like; an essence preparation such as a moisturizing essence, an anti-wrinkle essence, a skin repair essence, and the like; a gel preparation such as a moisturizing gel, an anti-wrinkle gel, a skin repair gel, and the like; a cream or a paste such as a face cream, an eye cream, a moisturizing cream, an anti-wrinkle cream, a skin repair cream, a massage cream, a hand cream, a body cream, and the like; a powder preparation such as a setting powder, a loose powder, a pressed powder, a body powder, and the like.

[0035] In addition to containing nerolidol as an effective ingredient for a skin barrier damage repair agent, the cosmetic having a skin barrier damage repair function of the present application can contain other additives such as an excipient, a carrier, and the like, and ingredients generally added in cosmetics can be appropriately used as needed.

[0036] Examples of the ingredients generally added in cosmetics include, but are not limited to, for example, an aqueous solvent, an oily ingredient, an anionic surfactant, a cationic surfactant, an amphoteric surfactant, a nonionic surfactant, a water-soluble polymer, a thickening agent, a preservative, a UV absorber, a metal ion blocking agent, an amino acid, an organic amine, a polymer emulsion, a pH adjustor, a neutralizer, a skin nutrient, a vitamin, an antioxidant, an antioxidant aid, a perfume, and the like.

[0037] Examples of the aqueous solvent include, for example, water, an alcohol, a humectant, or a mixture thereof.

[0038] As the water, water used in cosmetics, quasi drugs, and the like can be used, and for example, purified water, ion-exchanged water, tap water, and the like can be used. The aqueous phase can further contain a water-soluble alcohol according to the purpose.

[0039] Examples of the water-soluble alcohol include, for example, at least one selected from the group consisting of a lower alcohol, a polyhydric alcohol, a polyhydric alcohol polymer, a glycol alkyl ether, a glycol ether ester, a glycerol monoalkyl ether, a sugar alcohol, a monosaccharide, an oligosaccharide, a polysaccharide, and a derivative thereof, and the like.

[0040] Examples of the lower alcohol include, for example, ethanol, propanol, isopropanol, isobutanol, tert-butanol, and the like.

[0041] Examples of the polyhydric alcohol include, for example, a dihydric alcohol (for example, ethylene glycol, propylene glycol, trimethylene glycol, 1,2-butanediol, 1,3-butanediol, tetramethylene glycol, 2,3-butanediol, pentamethylene glycol, 2-buten-1,4-diol, hexanediol, octanediol, and the like); a trihydric alcohol (for example, glycerol, trimethylolpropane, and the like); a tetrahydric alcohol (for example, pentaerythritol, and the like); a pentahydric alcohol (for example, xylitol, and the like); a hexahydric alcohol (for example, sorbitol, mannitol, and the like).

[0042] As the polyhydric alcohol polymer, for example, diethylene glycol, dipropylene glycol, triethylene glycol, polypropylene glycol, tetraethylene glycol, diglycerol, triglycerol, tetraglycerol, and the like can be exemplified.

[0043] As the dihydric alcohol alkyl ether, for example, ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, ethylene glycol monophenyl ether, ethylene glycol monohexyl ether, ethylene glycol mono-2-methylhexyl ether, ethylene glycol isoamyl ether, ethylene glycol benzyl ether, ethylene glycol isopropyl ether, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, ethylene glycol dibutyl ether, and the like; diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, diethylene glycol monobutyl ether, diethylene glycol dimethyl ether, diethylene glycol diethyl ether, diethylene glycol butyl ether, diethylene glycol methyl ethyl ether, triethylene glycol monomethyl ether, triethylene glycol monoethyl ether, propylene glycol monomethyl ether, propylene glycol monoethyl ether, propylene glycol monobutyl ether, propylene glycol isopropyl ether, dipropylene glycol methyl ether, dipropylene glycol ethyl ether, dipropylene glycol butyl ether, and the like can be exemplified.

[0044] As the dihydric alcohol ether ester, for example, ethylene glycol monomethyl ether acetate, ethylene glycol monoethyl ether acetate, ethylene glycol monobutyl ether acetate, ethylene glycol monophenyl ether acetate, ethylene glycol dihexyl adipate, ethylene glycol disuccinate, diethylene glycol monoethyl ether acetate, diethylene glycol monobutyl ether acetate, propylene glycol monomethyl ether acetate, propylene glycol monoethyl ether acetate, propylene glycol monopropyl ether acetate, propylene glycol monophenyl ether acetate, and the like can be exemplified.

[0045] As the glycerol monoalkyl ether, for example, squiol, squiolin, and the like can be exemplified.

[0046] As the sugar alcohol, for example, sorbitol, maltitol, maltotriose, mannitol, sucrose, erythritol, glucose, fructose, amylolytic sugar, maltose, xylose, amylolytic sugar reduction alcohol, and the like; tetrahydrofurfuryl alcohol; POE-tetrahydrofurfuryl alcohol; POP-butyl ether; POP-POE-butyl ether; trioxa-propylene glyceryl ether; POP-glycerol ether; POP-glycerol ether phosphate; POP-POE-pentatricthanol ether, and the like can be exemplified.

[0047] As monosaccharides, for example, at least one selected from the group consisting of trioses (e.g., D-glyceraldehyde, dihydroxyacetone, etc.), tetroses (e.g., D-erythrose, D-erythrulose, D-threose, erythritol, etc.), pentoses (e.g., L-arabinose, D-xylose, L-lyxose, D-arabinose, D-ribose, D-ribulose, D-xylulose, L-xylulose, etc.), hexoses (e.g., D-glucose, D-talose, D- psicose, D-galactose, D-fructose, L-galactose, L-mannose, D-tagatose, etc.), heptoses (e.g., heptulose, etc.), octoses (e.g., octulose, etc.), deoxy sugars (e.g., 2-deoxy-D-ribose, 6-deoxy-L-galactose, 6-deoxy-L-mannose, etc.), amino sugars (e.g., D-glucosamine, D-galactosamine, sialic acid, muramic acid, etc.), uronic acids (e.g., D-glucuronic acid, L-guluronic acid, D-galacturonic acid, L-iduronic acid, etc.), and the like can be exemplified.

[0048] As oligosaccharides, for example, at least one selected from the group consisting of sucrose, umbelliferone, lactose, raffinose, α,α-trehalose, and the like can be exemplified.

[0049] As polysaccharides, for example, at least one selected from the group consisting of cellulose, buckthorn seed, chondroitin sulfate, starch, galactan, dermatan sulfate, glycogen, gum arabic, heparan sulfate, hyaluronic acid, gum tragacanth, keratan sulfate, chondroitin, xanthan gum, mucin sulfate, guar gum, dextran, locust bean gum, succinoglycan, carragenin, and the like can be exemplified.

[0050] As other polyols, for example, at least one selected from the group consisting of polyoxyethylene methyl glucoside (Glucam E-10), polyoxypropylene methyl glucoside (Glucam P-10), and the like can be exemplified.

[0051] As humectants, for example, propylene glycol, glycerin, 1,3-butanediol, dipropylene glycol, xylitol, sorbitol, maltitol, chondroitin sulfate, hyaluronic acid, mucin sulfate, carragenin, atelocollagen, sodium lactate, bile acid salts, dl-pyrrolidone carboxylate, alkylene oxide derivatives, short-chain soluble collagen, diglycerol (EO)PO adduct, roxburgh plum extract, yarrow extract, melilot extract, and the like can be exemplified.

[0052] As oily ingredients, for example, liquid oil, solid oil, wax, hydrocarbon, higher fatty acid, higher alcohol, synthetic ester oil, silicone oil, and the like can be used. Note that, in the present specification, oil components and components soluble in oil components are also included in the "oily ingredients".

[0053] As liquid fats and oils, for example, avocado oil, camellia oil, turtle oil, macadamia nut oil, corn oil, mink oil, olive oil, rapeseed oil, egg yolk oil, sesame oil, peach kernel oil (almond oil), wheat germ oil, camellia flower oil, castor oil, linseed oil, safflower oil, cottonseed oil, perilla oil, soybean oil, peanut oil, tea seed oil, torreya grandis oil, rice bran oil, Chinese tung oil, Japanese tung oil, jojoba oil, germ oil, triglycerin, and the like can be exemplified.

[0054] As solid fats and oils, for example, cocoa butter, coconut oil, horse oil, hydrogenated coconut oil, palm oil, beef tallow, mutton tallow, hydrogenated beef tallow, palm kernel oil, lard, beef bone fat (oil), candelilla oil, hydrogenated oil, beef foot oil, wood wax, hydrogenated castor oil, and the like can be exemplified.

[0055] As waxes, for example, beeswax, candelilla wax, cotton wax, carnauba wax, bayberry wax, insect wax (white wax), whale wax, montan wax, rice bran wax, lanolin, kapok wax, lanolin acetate, liquid lanolin, sugar cane wax, isopropyl lanolin fatty acid ester, hexyl laurate, reduced lanolin, jojoba wax, hard lanolin, shellac wax, POE lanolin alcohol ether, POE lanolin alcohol acetate, POE cholesterol ether, lanolin fatty acid polyethylene glycol ester, POE hydrogenated lanolin alcohol ether, and the like can be exemplified.

[0056] As hydrocarbon oils, for example, liquid paraffin, ozokerite, squalane, pristane, paraffin, pure ozokerite, squalene, vaseline, microcrystalline wax, and the like can be exemplified.

[0057] As higher fatty acids, for example, lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, undecylenic acid, tall oil acid, isostearic acid, linoleic acid, linolenic acid, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and the like can be exemplified.

[0058] As higher alcohols, for example, straight-chain alcohols (e.g., lauryl alcohol, cetyl alcohol, stearyl alcohol, behenyl alcohol, myristyl alcohol, oleyl alcohol, cetearyl alcohol, and the like); branched-chain alcohols (e.g., monostearyl glycerin ether (batyl alcohol), 2-decyltetradecyl alcohol, lanolin alcohol, cholesterol, phytosterol, hexyldodecyl alcohol, isostearyl alcohol, octyldodecyl alcohol, and the like); and the like can be exemplified.

[0059] As synthetic ester oils, there can be mentioned hydrogenated polydecene, isopropyl myristate, cetyl octanoate, octyldodecyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, decyl oleate, dimethyl octanoic acid hexyl decyl ester, cetyl lactate, myristyl lactate, lanolin acetate, isocetyl stearate, isocetyl isostearate, cholesteryl 12-hydroxystearate, ethylene glycol di-2-ethylhexanoate, dipentaerythritol fatty acid ester, N-alkyl glycol mono-isostearate, neopentyl glycol dicaprate, diisostearyl malate, glycerin tri-2- heptylundecanoate, trimethylolpropane tri-2-ethylhexanoate, trimethylolpropane triisostearate, pentaerythritol tetra-2-ethylhexanoate, glycerin tri-2-ethylhexanoate, glycerin trioctanoate, glycerin triisopalmitate, cholesteryl ethylhexanoate, 2-ethylhexyl palmitate, glycerin trimyristate, glycerin tri-2-heptylundecanoate, methyl ricinoleate, oleyl oleate, acetyl glycerin ester, 2-heptylundecyl palmitate, diisobutyl adipate, N-lauroyl-L-glutamic acid-2-octyldodecyl ester, di-2-heptylundecyl adipate, ethyl laurate, di-2-ethylhexyl sebacate, 2-hexyldecyl myristate, 2-hexyldecyl palmitate, 2-hexyldecyl adipate, diisopropyl sebacate, 2-ethylhexyl succinate, triethyl citrate, and the like.

[0060] As silicone oils, there can be mentioned dimethylpolysiloxane, methylhydrogenpolysiloxane, methylphenylpolysiloxane, stearyloxy methylpolysiloxane, polyether-modified organopolysiloxane, fluoroalkyl-polyoxyalkylene co-modified organopolysiloxane, alkyl-modified organopolysiloxane, end-modified organopolysiloxane, fluoro-modified organopolysiloxane, amino-modified organopolysiloxane, PEG-10 dimethylpolysiloxane, silica gel, acrylpolysiloxane, trimethylsiloxy silicate, silicone RTV rubber, and the like organosilicon compounds.

[0061] As the oil component to be used in combination with hydrogenated polyisobutylene, an oil component which is poorly compatible with hydrogenated polyisobutylene and which is volatile is preferred, so that in the application, the volatile oil component will volatilize, and due to the poor compatibility, the hydrogenated polyisobutylene will remain on the skin to form a film, resulting in a tightening effect.

[0062] As anionic surfactants, for example, the following can be listed: fatty acid soaps (e.g., sodium laurate, sodium palmitate, etc.); higher alkyl sulfate salts (e.g., sodium lauryl sulfate, potassium lauryl sulfate, etc.); alkyl ether sulfate salts (e.g., POE-lauryl sulfate triethanolamine, POE-lauryl sulfate sodium, etc.); N-acyl sarcosinates (e.g., sodium lauroyl sarcosinate, etc.); higher fatty acid amide sulfonates (e.g., sodium N-myristoyl-N-methyl taurate, sodium coconut oil fatty acid methyl taurate, sodium lauryl methyl taurate, etc.); phosphate ester salts (POE-oil ether phosphoric acid sodium, POE-stearyl ether phosphoric acid, etc.); sulfosuccinates (e.g., sodium di-2-ethylhexyl sulfosuccinate, sodium mono-lauroyl mono-ethanolamide polyoxyethylene sulfosuccinate, sodium lauryl polypropylene glycol sulfosuccinate, etc.); alkyl benzene sulfonate salts (e.g., sodium linear dodecyl benzene sulfonate, triethanolamine linear dodecyl benzene sulfonate, linear dodecyl benzene sulfonic acid, etc.); higher fatty acid ester sulfate salts (e.g., sodium hydrogenated coconut oil fatty acid glyceride sulfate, etc.); N-acyl glutamates (e.g., monosodium N-lauroyl glutamate, disodium N-stearyl glutamate, monosodium N-myristoyl-L-glutamate, etc.); sulfated oils (e.g., Turkey red oil, etc.); POE-alkyl ether carboxylic acids, POE-alkyl allyl ether carboxylic acid salts, alpha-olefin sulfonates, higher fatty acid ester sulfonates, secondary alcohol sulfates, higher fatty acid alkanolamide sulfates, sodium lauroyl monoethanolamide succinate, di-triethanolamine N-palmitoyl aspartate, sodium caseinate, potassium cetyl phosphate, etc.

[0063] As cationic surfactants, for example, the following can be listed: alkyl trimethyl ammonium salts (e.g., steartrimonium chloride, lauryltrimethyl ammonium chloride, etc.); alkyl pyridinium salts (e.g., cetyl pyridinium chloride, etc.); distearyldimethyl ammonium chloride, dialkyldimethyl ammonium chloride salts; poly(N,N"-dimethyl-3,5-methylene piperidinium) chloride; alkyl quaternary ammonium salts; alkyl dimethyl benzyl ammonium salts; alkyl isoquinolinium salts; dialkyl morpholinium salts; POE-alkyl amines; alkyl amine salts; polyamine fatty acid derivatives; amyl alcohol fatty acid ester derivatives; benzalkonium chloride; benzethonium chloride, etc.

[0064] As amphoteric surfactants, for example, the following can be listed: imidazoline-based amphoteric surfactants (e.g., 2-undecyl-N,N,N-(hydroxyethyl carboxymethyl)-2-imidazoline sodium, 2-cocoyl-2-imidazolinium hydroxide-1-carboxyethoxy disodium salt, etc.); betaine-based surfactants (e.g., 2-heptadecyl-N-carboxymethyl-N-hydroxyethyl imidazolinium betaine, lauryl dimethylaminoacetic acid betaine, alkyl betaine, amido betaine, sulfobetaine, etc.); etc.

[0065] As the lipophilic nonionic surfactant, for example, sorbitan fatty acid ester (e.g., sorbitan monooleate, sorbitan monoisostearate, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan sesquioleate, sorbitan trioleate, sorbitan penta-2-ethylhexanoate diglyceride, sorbitan tetra-2-ethylhexanoate diglyceride, etc.), polyglycerin fatty acid glyceride (e.g., monolinseed oil fatty acid glyceride, monoglyceride, sesqui-oleic acid glyceride, monostearic acid glyceride, α,α'-oleic acid pyroglutamic acid glyceride, glycerin malic acid monostearate, etc.), propylene glycol fatty acid ester (e.g., propylene glycol monostearate, etc.), hydrogenated castor oil derivative, glycerin alkyl ether, PEG-10 dimethicone, glycerin stearate, PPG-13-decyltetradecinol ether-24, PEG-5 glycerin stearate, etc. can be exemplified.

[0066] As the hydrophilic nonionic surfactant, for example, POE-sorbitan fatty acid ester (for example, POE-sorbitan monooleate, POE-sorbitan monostearate, POE-sorbitan monooleate, POE-sorbitan tetraoleate, and the like); POE-sorbitol fatty acid ester (for example, POE-sorbitol monolaurate, POE-sorbitol monooleate, POE-sorbitol pentaoleate, POE-sorbitol monostearate, and the like); POE-glycerin fatty acid ester (for example, POE-glycerin monostearate, POE-glycerin monoisostearate, POE-glycerin triisostearate, and the like POE-monooleate, and the like); POE-fatty acid ester (for example, POE-distearyl ester, POE-mono dioleate, ethylene glycol distearate, and the like); POE-alkyl ether (for example, POE-lauryl ether, POE-oil ether, POE-stearyl ether, POE-benzyl ether, POE-2-octyldodecyl ether, POE-cholestanol ether, and the like); Pluronic type (for example, Pluronic, and the like); POE·POP-alkyl ether (for example, POE·POP-spermium ether, POE·POP-2-decyl tetradecyl ether, POE·POP-monobutyl ether, POE·POP-hydrogen lanolin, POE·POP-glycerin ether, and the like); tetra-POE·tetra POP-ethylene diamine condensate (for example, Tetronic, and the like); POE-castor oil hydrogenated castor oil derivative (for example, POE-castor oil, POE-hydrogenated castor oil, POE-hydrogenated castor oil monoisostearate, POE-hydrogenated castor oil triisostearate, POE-hydrogenated castor oil monoglutamic acid monoisostearate diester, POE-hydrogenated castor oil maleate, and the like); POE-beeswax·lanolin derivative (for example, POE-sorbitol beeswax, and the like); alkanolamide (for example, coconut oil fatty acid diethanolamide, lauric acid monoethanolamide, fatty acid isopropyl alcohol amide, and the like); POE-propylene glycol fatty acid ester; POE-alkyl amine; POE-fatty acid amide; sucrose fatty acid ester; alkyl ethoxy dimethyl amine oxide; triolein phosphate; oleyl polyether-10, PEG-100 stearate, methoxy PEG / PPG-25 / 4 polydimethylsiloxane, polysorbate-60, PEG-40 stearate, sucrose stearate. As the natural water-soluble polymer, for example, plant-based polymer (for example, gum arabic, gum tragacanth, galactan, guar gum, carob gum, karaya gum, carrageenan, pectin, agar, quince seed, algal colloid (brown algae extract), starch (rice, corn, potato, wheat), glycyrrhizic acid), microorganism-based polymer (for example, xanthan gum, dextran, succinyl dextran, pullulan, and the like), animal-based polymer (for example, collagen, casein, albumin, gelatin, and the like), and the like can be exemplified.

[0067] As the semi-synthetic water-soluble polymer, for example, starch-based polymers (e.g., carboxymethyl starch, methylhydroxypropyl starch, etc.), cellulose-based polymers (methyl cellulose, ethyl cellulose, methylhydroxypropyl cellulose, hydroxyethyl cellulose, sodium cellulose sulfate, hydroxypropyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose, crystalline cellulose, cellulose powder, etc.), alginic acid-based polymers (e.g., sodium alginate, propylene glycol alginate, etc.), PEG-240 / HDI copolymer bis-dodecyltetradecinol polyether-20, acrylate / C10-30 alkyl acrylate cross-linked polymer, etc. can be listed.

[0068] As the thickening agent, for example, carbomer, acrylate / C10-30 alkyl acrylate cross-linked polymer, sodium acrylate / sodium acryloyldimethyltaurate copolymer, acrylate / hexadecyl ethoxy (20) itaconate copolymer, acrylate / hexadecyl ethoxy (20) methacrylate copolymer, acrylate / tetradecyl ethoxy (25) acrylate copolymer, acrylate / octadecyl ethoxy (20) itaconate copolymer, acrylate / octadecyl ethoxy (20) methacrylate copolymer, acrylate / octadecyl ethoxy (50) acrylate copolymer, acrylate / VA cross-linked polymer, PAA (polyacrylic acid), sodium acrylate / vinyl isodecanoate cross-linked polymer, Carbomer (polyacrylic acid) and its sodium salt, and other polyacrylic acid-based thickening agents can be listed.

[0069] As the preservative, for example, phenoxyethanol, benzyl alcohol, hydroxybenzoate, p-hydroxyacetophenone, and other aromatic preservatives can be listed.

[0070] As the ultraviolet absorber, for example, benzoic acid ultraviolet absorbers (e.g., p-aminobenzoic acid (hereinafter abbreviated as PABA), PABA monoglyceride, N,N-dipropoxy PABA ethyl ester, N,N-diethoxy PABA ethyl ester, N,N-dimethyl PABA ethyl ester, N,N-dimethyl PABA butyl ester, N,N-dimethyl PABA ethyl ester, and the like); anthranilic acid ultraviolet absorbers (e.g., N-acetylanthranilic acid homomenthyl ester, and the like); salicylic acid ultraviolet absorbers (e.g., amyl salicylate, menthyl salicylate, homomenthyl salicylate, octyl salicylate, phenyl salicylate, benzyl salicylate, p-isopropylphenyl salicylate, and the like); cinnamic acid ultraviolet absorbers (e.g., octyl methoxycinnamate, ethyl 4-isopropylcinnamate, methyl 2,5-diisopropylcinnamate, ethyl 2,4-diisopropylcinnamate, methyl 2,4-diisopropylcinnamate, propyl p-methoxycinnamate, isopropyl p-methoxycinnamate, isopentyl p-methoxycinnamate, octyl p-methoxycinnamate (2-ethylhexyl p-methoxycinnamate), 2-ethoxyethyl p-methoxycinnamate, cyclohexyl p-methoxycinnamate, ethyl α-cyano-β-phenylcinnamate, 2-ethylhexyl α-cyano-β-phenylcinnamate, glyceryl mono-2-ethylhexanoyl-dip-methoxycinnamate, and the like); benzophenone ultraviolet absorbers (e.g., 2,4-dihydroxybenzophenone, 2,2"-dihydroxy-4-methoxybenzophenone, 2,2"-dihydroxy-4,4"-dimethoxybenzophenone, 2,2",4,4"-tetrahydroxybenzophenone, 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxy-4"-methylbenzophenone, 2-hydroxy-4-methoxybenzophenone-5-sulfonate, 4-phenylbenzophenone, 2-ethylhexyl-4"-phenyl-benzophenone-2-carboxylate, 2-hydroxy-4-n-octoxybenzophenone, 4-hydroxy-3-carboxybenzophenone, and the like); 3-(4"-methylbenzylidene)-d,l-camphor, 3-benzylidene-d,l-camphor; 2-phenyl-5-methylbenzoxazole; 2,2"-hydroxy-5-methylphenylbenzotriazole; 2-(2"-hydroxy-5"-tert-octylphenyl)benzotriazole; 2-(2"-hydroxy-5"-methylphenylbenzotriazole; dibenzylhydrazine; dianisyl ether methane; 4-methoxy-4"-tert-butyl dibenzoylmethane; 5-(3,3-dimethyl-2- norbornylidene)-3-pentane-2-one, dimorpholinopyridazinone; 2-ethylhexyl-2-cyano-3,3-diphenylacrylate; 2,4-bis-{[4-(2-ethylhexyloxy)-2-hydroxy]-phenyl}-6-(4-methoxyphenyl)-(1,3,5)-triazine, and the like.

[0071] As the metal ion blocking agent, for example, 1-hydroxyethane-l, l-diphosphonic acid, 1-hydroxyethane-l, l-diphosphonic acid tetrasodium salt, disodium edetate (EDTA 2 sodium), trisodium edetate (EDTA 3 sodium), tetrasodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, phosphoric acid, citric acid, ascorbic acid, succinic acid, edetic acid, ethylenediamine hydroxyethyl triacetic acid trisodium, and the like can be exemplified.

[0072] As the amino acid, for example, neutral amino acids (e.g., threonine, cysteine, and the like), basic amino acids (e.g., hydroxylysine, and the like), and the like can be exemplified. In addition, as the amino acid derivative, for example, sodium acylmethionate (sodium lauroyl methionate), acyl glutamate, sodium acyl beta-alanine, glutathione, pyrrolidone carboxylic acid, and the like can be exemplified.

[0073] As the organic amine, for example, monoethanolamine, diethanolamine, triethanolamine, morpholine, triisopropanolamine, 2-amino-2-methyl-l, 3-propanediol, 2-amino-2-methyl-l-propanol, and the like can be exemplified.

[0074] As the high molecular emulsion, for example, acrylic resin emulsion, polyacrylate emulsion, acrylic resin solution, polyacrylate emulsion, polyvinyl acetate resin emulsion, natural rubber latex, and the like can be exemplified.

[0075] As the pH adjusting agent, for example, buffers such as lactic acid-sodium lactate, citric acid-sodium citrate, succinic acid-sodium succinate, and the like can be exemplified.

[0076] As the neutralizing agent, potassium hydroxide, sodium hydroxide, aminomethylpropanol, arginine, and the like can be exemplified.

[0077] As the vitamin, for example, vitamin A, B1, B2, B6, C, E, and derivatives thereof, pantothenic acid and derivatives thereof, biotin, and the like can be exemplified.

[0078] As the antioxidant, for example, tocopherols, dibutylhydroxytoluene, butylhydroxyanisole, gallate esters, and the like can be exemplified.

[0079] As the antioxidant aid, for example, phosphoric acid, citric acid, ascorbic acid, maleic acid, malonic acid, succinic acid, fumaric acid, cephalin, hexametaphosphate, phytic acid, ethylenediaminetetraacetic acid, and the like can be exemplified.

[0080] Examples

[0081] Hereinafter, the effect of nerolidol on the synthesis of lipids by human sebocytes will be further explained in detail through Test Examples and Examples, but the present application is not limited to the Test Examples and Examples described below.

[0082] Experimental Example 1. Verification of the effect of nerol on promoting lipid synthesis in SZ95 human sebaceous gland cells using Nile red fluorescent staining.

[0083] The effect of nerol on lipid synthesis in SZ95 human sebaceous gland cells was investigated using Nile Red fluorescence staining. SZ95 human sebaceous gland cells are an immortalized human sebaceous gland cell line that retains the morphological, phenotypic, and functional characteristics of normal human sebaceous gland cells and is often used as an in vitro alternative model for sebaceous gland biological research. Nile Red is a lipophilic fluorescent dye that binds to neutral lipids (ceryl esters, triglycerides, and various fatty acids, etc.) and emits a red fluorescent signal. After staining cells with Nile Red, the fluorescence intensity at excitation wavelengths of 490 nm and emission wavelengths of 565 nm was detected using a fluorescence microplate reader, flow cytometry, or confocal fluorescence microscopy, allowing for sensitive and reliable determination of intracellular neutral lipid levels.

[0084] SZ95 human sebaceous gland cells were processed at 2×10 4 Cells were seeded at a density of 96-well black transparent plates (WoHong Biotechnology, Shanghai, China) and cultured in a cell culture incubator for 24 h. Then, different concentrations of nerol (0-1000 μM) were added, and incubation was continued for 24 h (Figure 1, top) or 48 h (Figure 1, bottom). After incubation, the supernatant was discarded, and 100 μl of 10 μg / ml Nile Red solution (GLPBIO Technology Inc, USA) was added to each well. The plates were incubated for 30 min. After incubation, the cells were read using a fluorescence microplate reader. The fluorescence intensity of each well was detected using a Multi-detection system (Promega, Madison, WI, USA) at an excitation wavelength of 490 nm and an emission wavelength of 565 nm. The relative levels of intracellular neutral lipids in each experimental group were calculated based on the percentage of fluorescence intensity relative to the control group. The results are shown in Figure 1. The data in the figure are Mean ± SD from three replicate experiments. * p<0.05, ** p<0.01 vs blank control group.

[0085] The results in Figure 1 show that nerol significantly enhanced the level of neutral lipids in SZ95 sebaceous gland cells in a dose-dependent manner within the dose range of 250-1000 μM.

[0086] Experimental Example 2. Flow cytometry analysis of the effect of nerol on the fluorescence intensity of Nile Red-stained SZ95 cells.

[0087] To verify the effect of nerolidol on lipid synthesis in SZ95 human sebocytes, flow cytometry was used to analyze the effect of nerolidol on the fluorescence intensity of SZ95 cells dyed with Nile red. SZ95 human sebocytes were seeded in a 12-well cell culture plate (NEST, China) at a cell density of 1 x 10 5 cells / well in a cell culture incubator for 24 h, and then different concentrations of nerolidol (0, 250, 500, and 1000 μM) were added for 48 h. After the end of the culture, the cells were digested from the culture plate, resuspended in complete medium, centrifuged at 7,000 rpm at room temperature for 1 min, resuspended in phosphate buffered saline (PBS), and dyed with 10 μg / ml Nile red solution for 30 min. The fluorescence intensity of the cells in each experimental group was measured using a FACScalibur flow cytometer (BD Biosciences, San Jose, CA, USA) (excitation wavelength: 488 nm, emission wavelength: 550 nm). At least 1 x 10 4 cells of data were collected for each experimental group, and the flow cytometry data were analyzed using FlowJo V10 software. The relative level of neutral lipids in the cells in each experimental group was calculated based on the percentage of the fluorescence intensity of each experimental group relative to the control group. The results are shown in FIG. 2. The upper panel is a histogram of the fluorescence intensity of each experimental group analyzed by flow cytometry, which is a representative result of three independent repeated experiments. The lower panel is the quantitative analysis result of the lipid level in the cells. The data in the figure are Mean ± SD from three repeated experiments. * p<0.05, ** p<0.01 vs blank control group.

[0088] The results of FIG. 2 show that nerolidol (250-1000 μM) can significantly increase the lipid level in SZ95 human sebocytes in a dose-dependent manner.

[0089] Test Example 3. Test for detecting the effect of nerolidol on lipid droplet accumulation in SZ95 cells using confocal fluorescence microscopy

[0090] The effect of nerolidol on lipid droplet accumulation in SZ95 cells was detected using confocal fluorescence microscopy to further verify the effect of nerolidol on lipid synthesis in SZ95 human sebocytes. SZ95 human sebocytes were seeded in a 12-well cell culture plate (NEST, China) at a cell density of 4 x 10 5The cells were seeded at a cell density of 2×10 cells / well in BeyoGold™ 35 mm glass-bottom confocal dishes (Beyotime Biotechnology Co., Ltd., Shanghai, China) and incubated in a cell culture incubator for 24 h, after which different concentrations of nerolidol (0, 250, 500, 1000 mM) were added for 48 h. At the end of the treatment, the cells were washed with PBS and fixed with 4% formaldehyde for 15 min at room temperature. The cells were then stained with 10 pg / ml of Nile red solution for 15 min and counterstained with DAPI (1:1000, Sigma Aldrich, USA) for the nuclei. Images of the stained cells were taken using a STELLARIS 8 STED super-resolution confocal microscope (Leica, Germany), and the results are shown in Figure 3, which are representative of three independent experiments.

[0091] The results of Figure 3 show that nerolidol (250-1000 mM) can significantly increase the accumulation of lipid droplets in SZ95 human sebocytes in a dose-dependent manner.

[0092] The experimental results of Test Examples 1-3 (Figures 1-3) above show that nerolidol has the effect of promoting the synthesis of lipids by human sebocytes.

[0093] Next, the effect of nerolidol on the differentiation of human sebocytes was verified.

[0094] Lipid synthesis is an important feature of sebocyte differentiation, and therefore, the promotion of lipid synthesis by SZ95 human sebocytes by nerolidol reflects its possible effect of inducing the differentiation of sebocytes. In addition to lipid synthesis, the loss of cell proliferation ability, the increase in cell granulation, and the expression of differentiation marker genes such as epithelial membrane antigen (EMA), keratin-7 (keratin 7), and peroxisome proliferators-activated receptor-gamma (PPARy) are also markers of sebocyte differentiation. To further confirm the effect of nerolidol on the differentiation of human sebocytes, Test Examples 4-6 detected the effects of nerolidol on the proliferation, granulation, and expression of differentiation marker genes such as EMA, keratin 7, and PPARy of SZ95 human sebocytes.

[0095] Test Example 4. Test on the effect of nerolidol on the inhibition of the proliferation of SZ95 human sebocytes

[0096] First, the inhibitory effect of nerolidol on the proliferation of SZ95 human sebocytes was verified by a CCK8 (Cell Counting Kit-8) experiment. SZ95 human sebocytes were seeded at a cell density of 2×104 The cells were seeded in 96-well cell culture plates at a cell density of 5000 cells / well, and incubated in a cell incubator for 24 h. After the incubation, different concentrations of neroli (0, 125, 250, 500, 1000 μM) were added, and incubated for 24 h or 48 h. The supernatant was discarded, and the cells were washed twice with PBS. Then, 100 μl of CCK-8 solution (100 μl of CCK-8 reagent was added per ml of culture medium) was added to each well. The plates were incubated in the incubator for 1 h. After the incubation, the optical density (OD) value at 450 nm of each well was detected by an enzyme-labeled instrument (BioTek Epoch Co., Ltd., Winooski, VT, USA). The proliferation rate of SZ95 human sebaceous gland cells was calculated according to the percentage of the OD value of each experimental group relative to the OD value of the control group. The results are shown in FIG. 4, and the data in the figure are from Mean ± SD of three independent repeated experiments. ** p < 0.01 vs. the blank control group.

[0097] The results of FIG. 4 show that neroli has no obvious effect on the viability and proliferation of SZ95 human sebaceous gland cells in the dose range of 125-1000 μM for 24 h, but can significantly inhibit the proliferation of SZ95 human sebaceous gland cells in the dose range of 500-1000 μM for 48 h.

[0098] Test Example 5. Test on the effect of neroli on improving the granularity of SZ95 human sebaceous gland cells

[0099] When the cells pass through the laser detection area of the flow cytometer, light scattering occurs, and the direction of light scattering is related to the size and granularity of the cells. Forward scattering light (FS) is related to the size of the cells, and side scatter light (SS) is proportional to the granularity of the cells. The FACScalibur flow cytometer was used to measure the side scatter light intensity of SZ95 human sebaceous gland cells after being treated with different concentrations of neroli (0, 500, 1000 μM) for 24 h. The results are shown in FIG. 5, which is the flow cytometry histogram of the side scatter light of each experimental group, and is a representative experimental result of three independent repeated experiments.

[0100] The results of FIG. 5 show that the flow cytometry histogram of the side scatter light of SZ95 human sebaceous gland cells is obviously shifted to the right after being treated with neroli (500, 1000 μM) for 24 h, indicating that the granularity of the cells is obviously improved after being treated with neroli.

[0101] Test Example 6. Test on the effect of neroli on inducing SZ95 human sebaceous gland cells to express differentiation marker genes

[0102] Real-time quantitative PCR (qPCR) was used to detect the effect of nerol on the expression of differentiation marker genes EMA, keratin 7 and PPARy in SZ95 human sebocytes. SZ95 cells were seeded in 6-well cell culture plates at a cell density of 2 x 10 5 cells / well, and incubated in a cell incubator for 24 h. Different concentrations of nerol (0, 500, 1000 μM) were then added, and incubated for another 24 h. The mRNA expression levels of EMA, keratin 7 and PPARy in the cells were then determined by qPCR. Total RNA was extracted from the SZ95 cells using an RNA extraction kit (Yixing Biotech (Shanghai) Co., Ltd.), and 1 μg of the total RNA was reverse transcribed into cDNA using a PrimeScript RT kit (Hunan Aikangrui Biological Engineering Co., Ltd.). PCR was performed on a Quantum Studio 6 real-time PCR system (Thermo Fisher Scientific, Inc., Waltham, MA, USA) using SYBR Green Master Mix (Hunan Aikangrui Biological Engineering Co., Ltd.). The sequences of the primers used were as follows: EMA: forward: 5'-GTG CTG GTC TGT GTT CTG GTT G-3', reverse: 5'-ACT CGC TCA TAG GAT GGT AGG TAT C-3'; keratin 7: forward: 5'-TTG TGG TGC TGA AGA AGG ATG TG-3', reverse: 5'-GCT CTG TCA ACT CCG TCT CAT TG-3'; PPARy: forward: 5'-TGA ATC CAG AGT CCG CTG ACC TC-3', reverse: 5'-ATC GCC ATC GCC TTT GCT TTG-3'. The mRNA relative levels of each target gene were quantified using 2- ΔΔCt The results are shown in FIG. 6, which shows Mean ± SD from three independent repeated experiments. * p < 0.05, ** p < 0.01 vs blank control group.

[0103] The results of FIG. 6 show that the mRNA expression levels of EMA, keratin 7 and PPARy in the SZ95 human sebocytes were significantly increased after the cells were treated with nerol (500, 1000 μM) for 24 h, indicating that nerol has the effect of inducing the expression of differentiation marker genes in sebocytes.

[0104] The experimental results of Test Examples 4-6 (Figs. 4-6) show that nerolidol can inhibit the proliferation of SZ95 human sebocytes, increase the granularity of the cells, and induce the expression of differentiation marker genes such as EMA, keratin 7, and PPARγ, further verifying the effect of nerolidol on inducing the differentiation of human sebocytes.

[0105] Next, the effect of nerolidol on promoting the expression of transcription factors and fatty acid synthase involved in the regulation of sebum production is verified.

[0106] Peroxisome proliferators-activated receptors (PPARs) are a class of ligand-activated transcription factors involved in the regulation of the expression of multiple genes related to lipid metabolism. PPARγ is the most widely studied PPAR subtype, and its role in regulating sebum metabolism has been confirmed. Some studies have reported that PPARγ increases lipid accumulation in rat and human sebocytes, and patients treated with thiazolidinediones, a PPARγ agonist, have significantly increased sebum production. Sterol regulatory element-binding protein-1 (SREBP-1) is a "master regulator of lipid homeostasis". The newly synthesized SREBP-1 precursor protein in the cell is inactive and is bound to the endoplasmic reticulum membrane. When the cell is stimulated by lipogenic stimuli, SREBP-1 is proteolytically released to form an active protein, which enters the nucleus and binds to the sterol regulatory element (SRE) in the corresponding target gene promoter / enhancer region, promoting the expression of downstream PPARγ and lipid synthesis enzyme genes. Fatty acid synthase (FAS) is a key enzyme for fatty acid synthesis, which catalyzes the formation of long-chain fatty acids from acetyl-CoA and malonyl-CoA. The gene expression of FAS is regulated by PPARγ and SREBP-1. In order to further verify the effect of nerolidol on promoting the synthesis of lipids by sebocytes and to explore its intracellular mechanism, the present application uses western blotting to detect the effect of nerolidol on the expression levels of FAS, SREBP-1, and PPARγ proteins in SZ95 human sebocytes.

[0107] Test Example 7. Test for detecting the effect of nerolidol on the expression level of FAS, SREBP-1 and PPARy protein in SZ95 human sebocytes by western blotting method

[0108] SZ95 human sebocytes were seeded in 6-well cell culture plates at a cell density of 2 x 10 5 cells / well in a cell culture incubator for 24 h, and then different concentrations of nerolidol (0, 500, 1000 μM) were added and incubated for 24 h. After that, the cells were collected and lysed with cell lysis buffer (Beyotime, China), and the supernatant was collected by centrifugation. The protein content was determined by BCA method, and 20 μg of protein sample was loaded onto SDS-PAGE for electrophoretic separation. After that, the separated protein bands were transferred to a nitrocellulose membrane, which was blocked with 5% BSA-TBST. Then, the primary antibodies against SREBP-1 (Abeam, UK), FAS and PPARy (Cell Signaling Technology, Danvers, USA) were incubated overnight, followed by incubation with horseradish peroxidase-labeled secondary antibodies. The chemiluminescent substrate (Vazyme, China) was used for development, and the protein blot image was taken by Tanon multifunctional imager (Tanon, China). The experimental results are shown in Figure 7.

[0109] The results of Figure 7 show that nerolidol (500, 1000 μM) can significantly increase the protein levels of FAS, SREBP-1 and PPARy in SZ95 human sebocytes after 24 h of incubation, indicating that nerolidol promotes the synthesis of sebum in sebocytes by inducing the expression of transcription factors such as SREBP-1 and PPARy and lipid synthesis enzymes such as FAS.

[0110] Example 1. Cosmetic water

[0111] According to the following formula, a cosmetic water containing nerolidol as a skin barrier damage repair active ingredient was prepared by a conventional cosmetic manufacturing process.

[0112] Example 2. Cosmetic water

[0113] According to the following formula, a cosmetic water containing nerolidol as a skin barrier damage repair active ingredient was prepared by a conventional cosmetic manufacturing process.

[0114] Example 3. Emulsion

[0115] According to the following formula, an emulsion containing nerolidol as a skin barrier damage repair active ingredient was prepared by a conventional cosmetic manufacturing process.

[0116] Example 4. Emulsion

[0117] An emulsion containing nerolidol as a skin barrier damage repair active ingredient was prepared according to the following formulation using conventional cosmetic manufacturing procedures.

[0118] Example 5. Cream

[0119] A cream containing nerolidol as a skin barrier damage repair active ingredient was prepared according to the following formulation using conventional cosmetic manufacturing procedures.

[0120] Example 6. Cream

[0121] A cream containing nerolidol as a skin barrier damage repair active ingredient was prepared according to the following formulation using conventional cosmetic manufacturing procedures.

[0122] Example 7. Hydrogel

[0123] A hydrogel containing nerolidol as a skin barrier damage repair active ingredient was prepared according to the following formulation.

[0124] An appropriate amount of deionized water was heated to 85°C in a water bath, and Carbomer U20 powder was slowly added to the water and stirred until dissolved. The solution was then neutralized with 10% arginine. A mixture of p-hydroxyacetophenone and 1,2-hexanediol was dissolved in an appropriate amount of deionized water, and then added to the Carbomer U20 gel. The resulting mixture was stirred until homogeneous, and then cooled to below 40°C. Nerolidol was then added and stirred until homogeneous, resulting in a nerolidol hydrogel.

[0125] Example 8. Hydrogel

[0126] A hydrogel containing nerolidol as a skin barrier damage repair active ingredient was prepared according to the following formulation using the manufacturing method described in Example 7.

[0127] Industrial applicability

[0128] Orange flower alcohol has the effect of promoting the differentiation and lipid synthesis of human sebocytes, and can induce the expression of transcription factors regulating sebum production such as PPARγ, SREBP-1, and lipid synthesis enzymes such as FAS. Since hyposebaceous function and reduced sebum secretion can lead to skin barrier dysfunction, dry skin, and even be associated with accelerated skin aging and the development of certain skin diseases (such as AD, senile pruritus, and senile sebaceous deficiency eczema), orange flower alcohol is likely to have beneficial effects on hyposebaceous function populations (including AD patients and dry skin elderly) through its effect of promoting the differentiation and lipid synthesis of sebocytes, and can be used for the prevention and treatment or relief of dry skin, premature skin aging, AD, senile pruritus, and senile sebaceous deficiency eczema associated with reduced sebum secretion.

[0129] According to the present application, a new sebocyte lipid synthesis promoter can be provided, and using the new sebocyte lipid synthesis promoter, a skin barrier damage repair agent can be prepared, and a cosmetic with skin barrier damage repair function can be prepared, and thus a cosmetic for repairing at least one of skin barrier damage caused by hyposebaceous function and / or reduced sebum secretion, dry skin, premature skin aging, skin aging, skin pruritus, and senile sebaceous deficiency eczema can be developed.

Claims

1. A promoter of lipid synthesis by sebocytes, characterized in that, containing nerolidol.

2. Use of nerolidol in the manufacture of an agent for promoting the synthesis of lipids by sebocytes.

3. Use of nerolidol in the manufacture of an agent for repairing a skin barrier damage, the skin barrier damage being a skin barrier damage resulting from a decrease in sebaceous gland functionality and / or a decrease in sebaceous secretion.

4. Use of nerolidol in the manufacture of a cosmetic for promoting the repair of a skin barrier, the skin barrier being a skin barrier resulting from a decrease in sebaceous gland functionality and / or a decrease in sebaceous secretion.

5. Use according to claim 3 or 4, the skin barrier damage being at least one of a skin dryness, a skin premature aging, a skin aging, a skin pruritus, a senile sebaceous deficiency eczema resulting from a decrease in sebaceous gland functionality and / or a decrease in sebaceous secretion.

Images