CN114224786A

CN114224786A - Composition comprising ceramide and uses thereof

Info

Publication number: CN114224786A
Application number: CN202111527007.5A
Authority: CN
Inventors: 殷庆飞; 陈媛祺; 张瑶瑶; 郭奕光; 贾海东
Original assignee: Shanghai Jahwa United Co Ltd
Current assignee: Shanghai Jahwa United Co Ltd
Priority date: 2021-12-14
Filing date: 2021-12-14
Publication date: 2022-03-25

Abstract

The invention provides a grease composition containing ceramide, wherein the grease is selected from the following components: vegetable fat and oil, animal fat and oil or a combination thereof, wherein the vegetable fat and oil is selected from the following components: shea butter, olive oil, safflower seed oil, meadowfoam seed oil, plant squalane, or a combination thereof, wherein the animal fat is deep sea fish oil, wherein the composition comprises 0.01 to 5 wt% ceramide, based on the total weight of the composition. The invention also relates to application of the oil composition containing ceramide in enhancing the skin barrier function and/or in preparing medicines and/or skin external preparations for enhancing the skin barrier function.

Description

Composition comprising ceramide and uses thereof

Technical Field

The invention belongs to the field of cosmetics, and particularly relates to a ceramide-containing grease composition and related application thereof in repairing skin barriers.

Background

The healthy skin barrier can resist the entrance of external harmful substances, irritants and sunlight, and has the effects of moisturizing and regulating. Having a healthy skin barrier is equivalent to having beautiful natural skin. However, the skin barrier is damaged due to environmental, dietary and wrong skin care, so that the self-defense ability of the skin is insufficient and the skin is extremely sensitive and damaged. The state of skin hydration is extremely important for barrier repair, and it is therefore necessary to keep the skin constantly moist regardless of whether it is water or cream, but for dry and extremely dry skin, an outermost oily protective film is essential.

The vegetable oil and the mineral oil are common matrix raw materials of skin care products such as cream, emulsion and the like, and simultaneously have the moisturizing effect, and the action mechanism is that a hydrophobic thin-layer oil film is formed on the surface of the skin as a sealant function to prevent or delay the evaporation and the loss of water. In addition, some animal oil is applied to cosmetics, can be permanently acted on skin to enable the skin to feel soft and smooth, and reduce the loss of skin moisture, but the molecular weight of the animal oil is large, so that the animal oil is not easily absorbed by the skin, is relatively greasy, and is not easy to use for multiple purposes. The natural vegetable oil has good permeability to skin, contains a large amount of squalene, vitamin A, D, E, F, K, phytosterol and other skin-required nutrients, and has important effects on keeping skin healthy, promoting skin moisture content, repairing skin barrier, improving skin elasticity and resisting aging. For example, in the literature, "the effects of vegetable oil and mineral oil on the moisture content of skin and the water loss rate through the skin" is compared, "and it is proposed in the ninth conference on the science of cosmetics and academic research that the effects of vegetable oil and mineral oil on the water loss rate through the skin and the epidermis are not greatly different, but the moisture content of vegetable oil is higher than that of mineral oil in a short period of time, so that the moisturizing effect of vegetable oil is better than that of mineral oil, which is related to the fact that vegetable oil contains a large amount of moisturizing effective components such as unsaturated fatty acid, lecithin, squalene and phytosterol. For example, the paper sunflower, "the comprehensive effect of the mixed vegetable oil and the white oil on the skin is compared", the perfume and essence cosmetics [ J ].2013, 6(3):27-32 indicate that the vegetable oil shows excellent moisturizing effect on the skin no matter the vegetable oil is used for a short time or a long time, can effectively improve the moisture content of the skin and reduce the moisture loss of the skin, and can improve the elasticity of the skin and effectively reduce the wrinkle area, length and depth of the skin after the vegetable oil is used for a long time; white oil has certain improvement effect on the moisture content and moisture loss of the skin in a short term and a long term, but has the effect of reducing the wrinkle depth of the skin after long-term use. The comparison shows that the vegetable oil has better skin aging prevention and care effects than the white oil.

The skin barrier, generally referred to as the physical barrier of the skin, includes the stratum corneum layer of the epidermis of the skin and the sebaceous membrane located on the surface of the stratum corneum layer. The stratum corneum is located at the outermost layer of the epidermis and has a "brick-wall structure", wherein "bricks" refer to keratinocytes and "mortar" refers to intercellular lipids. In the process of skin barrier formation, more than 200 genes are highly associated with the skin barrier. For example, the barrier factors involved in the formation of the cornified envelope, silk polyprotein (FLG), Loricrin (LOR), endothelin (IVL), transglutaminase 1(TGM1), and the barrier moisturizing-related factors caspase 14(CASP14), HMGCoA reductase (HMGCR), aquaporin 3(AQP3), and the keratinocyte intercellular junction-related factor zonulin 1 (ZO-1).

In particular, Filaggrin (FLG) is an important constituent protein of the cornified envelope of the outer layer of the epidermis of the skin, mainly present in the stratum granulosum and stratum lucidum, and an important constituent structure of the skin barrier; and can be further degraded into hydrophilic free amino acids and water-absorbing derivatives, which can help skin store water and maintain skin water content, and has moisturizing effect. Loricrin (LOR) is a major component of the cornified envelope and plays an important role in the barrier function of the epidermis. Endothelin (IVL) is a marker protein for keratinocyte differentiation, mainly expressed in the upper part of the acanthocyte layer and the granular layer, and is located in the outer layer of the cornified envelope, and is covalently bound to a ceramide containing a hydroxyl group — OH, thereby linking the lipid matrix and the keratinocytes to each other to function. The characteristic resistance and insolubility of the cornified envelope is based on the very stable isopeptide bond formation catalyzed by transglutaminase 1(TGM 1).

HMGCoA reductase (HMGCR) is a rate-limiting enzyme for synthesizing cholesterol, exists in endoplasmic reticulum, catalyzes and synthesizes methyl dihydroxy pentanoic acid, then generates squalene through multi-step reactions such as decarboxylation, phosphorylation and the like, and finally converts the squalene into cholesterol after cyclization. Aquaporin 3(AQP3) is the most abundant aquaporin subtype expressed by AQPs family, has spatial hierarchy in the expression quantity of epidermis, and is mainly expressed in the basal layer, the acanthocyte layer and the granular layer of human epidermis and gradually disappears to the stratum corneum. This spatial distribution is consistent with the moisture content distribution of the skin: the water content of the basal layer and the upper part of the basal layer is about 75%, while the stratum corneum is only about 10% -15%. The high expression of AQP3 in the basal lamina membranacea can promote the transportation of water, glycerol and urea, so that the extracellular environment of the basal lamina membranacea is closer to a neutral equilibrium state; the closer to the stratum corneum, the less AQP3 was expressed, the more the water content decreased significantly, and the environment in the skin became increasingly acidic. In addition, AQP3 plays an important role in the transport of keratin glycerol, and AQP3 transports endogenous glycerol and triglycerides in sebaceous glands into the epidermis, so that it directly or indirectly affects the skin moisturizing effect.

Tight junctions are an important component of the physical barrier, mostly present in the epidermal layer of the skin, laterally at the top of the adjacent intercellular spaces. Tight junctions are composed of different types of transmembrane and intracellular cytoplasmic proteins. Transmembrane proteins such as sluice protein 1(CLDN1), Occludin (OCLN), etc.; cytoplasmic proteins include mainly zonula occludens-1 (ZO-1). ZO-1 is a cytoplasmic protein belonging to the family of membrane-associated guanylate kinase proteins, which aggregates atresia and sluice proteins via extracellular signaling pathways to form a basic cell junction band; ZO-1 is a cytoplasmic protein most characteristic of skin barrier function.

Plant biomimetic lipid technology (Phyto Bionic Sebum, PBS technology for short): a plurality of natural vegetable oils are selected, natural lipid components in a skin barrier structure are simulated through scientific proportioning, unsaturated fatty acids such as linolenic acid, linoleic acid and the like which are necessary for a skin cuticle are supplemented, lipid components lacking in the skin are supplemented, and the lipid components are compatible with sebum, so that the skin barrier is repaired quickly, and the skin is enabled to be in a healthy, moist and glossy state.

Wherein: the vegetable oil and fat is mainly derived from, but not limited to, plant fruits (e.g., shea butter, olive oil, almond oil, avocado oil, sea buckthorn oil, coconut oil, orange oil, bergamot oil, citron oil, tomato oil, rosa canina oil, piper oil, brazil nut oil, vanilla oil, anise oil, ruined palm oil, juniper berry oil, wenzhou mandarin fruit oil, star palm fruit oil, wild rose fruit oil, and coriander fruit oil), seeds (e.g., jojoba seed oil, safflower seed oil, meadowfoam nut oil, sunflower seed oil, macadamia, castor bean seed oil, peony seed oil, grape seed oil, apple seed oil, pomegranate seed oil, pumpkin seed oil, linseed oil, rapeseed oil, sesame seed oil, glass seed oil, brazil nut oil, berba seed oil, etc.), sesame seed oil, olive seed oil, apricot seed oil, olive seed oil, avocado seed oil, olive, Kapok seed oil, tea seed oil, Sichuan valley seed oil, papaya seed oil, raspberry seed oil, Adonis amurensis seed oil, calendula seed oil, bitter tree seed oil, moringa oleifera seed oil, pecan seed oil, perilla seed oil, babassu seed oil, white lupin seed oil, and the like) and germ (for example: rice germ oil, wheat germ oil, oat germ oil, corn germ oil, etc.), and also other parts such as leaves, barks, roots, petals, pistils, etc., which are partially derived from plants.

The invention unexpectedly discovers that the ceramide-containing oil and fat composition has skin barrier-related efficacy, can improve the skin barrier activity and promote the expression of skin barrier-related factors FLG, LOR, IVL, TGM1, CASP14, HMGCR, AQP3 and ZO-1. Therefore, the oil and fat composition containing ceramide can be used as an efficacy additive in medicines and skin external preparations.

Disclosure of Invention

In one aspect, the present invention provides a fat composition comprising ceramide, wherein the fat is selected from the group consisting of: vegetable fat and oil, animal fat and oil or a combination thereof, wherein the vegetable fat and oil is selected from the following components: shea butter, olive oil, safflower seed oil, meadowfoam seed oil, plant squalane, or a combination thereof, wherein the animal fat is deep sea fish oil, wherein the composition comprises 0.01 to 5 wt% ceramide, based on the total weight of the composition.

In a preferred embodiment, the composition of the invention further comprises a natural surfactant. In a preferred embodiment, the natural surfactant is selected from the group consisting of: lecithin, cholesterol, lanolin, tea saponin, protein, saponins, saccharides, alkyl polyglycoside or their combination. In a more preferred embodiment, the natural surfactant is hydrogenated lecithin.

In a preferred embodiment, the weight ratio of oil to natural surfactant in the composition of the invention is from 1:1 to 30: 1. In a more preferred embodiment, the weight ratio of oil to natural surfactant in the composition of the invention is from 10:1 to 30: 1.

In another aspect, the present invention also relates to the use of a ceramide-containing lipid composition for enhancing the barrier function of the skin. In a preferred embodiment, the enhancement of the skin barrier function is achieved by increasing the skin barrier activity and/or promoting the expression of skin barrier-associated factors.

In another aspect, the present invention relates to the use of an oil or fat composition comprising ceramide for the preparation of a medicament and/or a skin external preparation for enhancing the barrier function of the skin. In a preferred embodiment, the pharmaceutical product and/or the external preparation for skin comprises 0.001 to 100% by weight of the composition.

Detailed Description

The invention relates to an oil and fat composition containing ceramide and application thereof, and the composition is found to have the effects of improving skin barrier activity and promoting the expression of skin barrier related factors FLG, LOR, IVL, TGM1, CASP14, HMGCR, AQP3 and ZO-1 in research. Therefore, the oil and fat composition containing ceramide can be used as an efficacy additive for preparing medicines and/or skin external preparations for enhancing the skin barrier function.

To provide a more concise description, some of the quantitative representations presented herein are not modified by the term "about". It is understood that each quantity given herein is intended to refer to the actual given value, regardless of whether the term "about" is explicitly used, and also to refer to the approximation to such given value that would reasonably be inferred by one of ordinary skill in the art, including approximations due to experimental and/or measurement conditions for such given value.

To provide a more concise description, some quantitative expressions are recited herein as a range from about an X amount to about a Y amount. It should be understood that when a range is recited, the range is not limited to the upper and lower limits recited, but includes the entire range from about the X amount to about the Y amount or any amount therebetween.

Oil and fat

The grease composition of the present invention may contain various types of grease.

In some embodiments, the compositions of the present invention comprise a vegetable fruit oil. The plant fruit oil in the invention is also called plant fruit oil, which is mainly from plant fruits, seeds and embryos, and also has parts from other parts of plants such as leaves, barks, roots, petals, stamens and the like. Vegetable fruit oils include, but are not limited to, Butyrospermum parkii (Butyrospermum parkii) fruit fat (also known as shea butter), Olea europaea (Olea europaea) fruit oil (also known as olive oil), apricot (Prunus armeniaca) kernel oil (also known as sweet almond oil), avocado (Persea gratisma) fruit oil, Hippophae rhamnoides (Hippophae rhamnoides) fruit oil, Cocos nucifera fruit oil, Citrus junos (Citrus junos) fruit oil, Citrus bergamia (Citrus aurantium bergamia) fruit oil, Citrus medica (Citrus bergamia) fruit oil, Lycopersicon esculentum (Solomonisco sium) fruit oil, Citrus canina (Rosa canina) fruit oil, Pepper (Pipernium) fruit oil, Myrothecium brazianum (Carriera) fruit oil, Citrus depressa (Pimpinella) fruit oil, Citrus depressa officinalis fruit oil, Citrus depressa (Pimpinella) fruit oil, Citrus depressa (Pimpinella) fruit oil, and Pimpinella oil, Fruit oil of Rosa multiflora (Rosa multiflora) and fruit oil of Coriandrum sativum (Coriandrum sativum).

For example, shea butter has indexes close to those of sebum secretion oil of human body, contains rich non-saponified components, is easy to be absorbed by human body, can prevent drying and cracking, further recovers and maintains the natural elasticity of skin, protects the barrier function of skin, and also has the function of diminishing inflammation. The olive oil has a slight skin feel, is very suitable for dry skin, contains squalene and fatty acid which can be quickly absorbed by the skin, effectively keeps the skin elastic and moist, contains abundant monounsaturated fatty acid, vitamin and phenolic substances, can eliminate facial wrinkles, prevents skin aging, and has the effects of protecting skin and hair, preventing rhagadia manus et pedis and the like. The sweet almond oil can soften and condition skin, has good repairing and moisturizing effects on dry, cracked, itchy or irritated skin, and is particularly suitable for dry and sensitive skin. Avocado oil is used as a cosmetic lubricant, and has effects in smoothing skin, promoting skin to be smooth, promoting skin cell regeneration, and repairing sunburn and dry skin. Coconut oil is saturated fat, has stable lipid, is not easy to oxidize to generate free radical attack, has strong antioxidation capability to help human body to prevent the generation of free radicals, can be used for beautifying and protecting skin, has emulsification stabilizing effect and antioxidation, and can make cosmetics more uniform and fine.

In some embodiments, the compositions of the present invention comprise 1 to 50% by weight of vegetable fruit oil. In a preferred embodiment, the composition of the invention comprises 40-50 wt% of vegetable fruit oil.

In some embodiments, the compositions of the present invention comprise a plant seed oil. The plant seed oil used in the present invention is also called plant seed oil, including but not limited to: jojoba seed oil, safflower seed oil, meadowfoam seed oil and sunflower seed oil. The oil and fat containing plant seed mainly includes jojoba (Simmondsia chinensis) seed oil, safflower (Carthamus tinctorius) seed oil, Potentilla chinensis (Limnanthes alba) seed oil, sunflower (Helianthus annuus) seed oil, Macadamia ternifolia (Macadamia ternifolia) seed oil, Ricinus communis (Ricinus communis) seed oil, peony (Paeonia suffruticosa) seed oil, grape (Vitisia vinifera) seed oil, grape fruit (Citrus paradisi) seed oil, apple (Pyrus malus) seed oil, pomegranate (Punica grantum) seed oil, watermelon (Citrullus lanatus) seed oil, pumpkin (Cucura peltata) seed oil, flax (Linum usitatissimum) seed oil, rape (Brassica campestris) seed oil, sesame (Sempm sambucinum) seed oil, glass papaya (Borax grandis) seed oil, Caryopteria mangium sativum seed oil, papaya seed oil (Acacia mangium seed oil), Raphus sativum seed oil, Raphus chinensis seed oil, Rubus sativus seed oil, Raphus sativus seed oil, and Caryopteri seed oil, Monkey tree (Adansonia digita) seed oil, Calendula officinalis (Calendala officinalis) seed oil, bitter tree (Carapa guaianensis) seed oil, Moringa oleifera (Moringa oleifera) seed oil, Carya illinoinensis (Carya illinoinensis) seed oil, Perilla frutescens (Perilla octoides) seed oil, Babesia babassu (Orbignya oleica) seed oil, Lupinus albus (Lupinus albus) seed oil, and the like.

For example, safflower oil is rich in linoleic acid and fat-associated substances (vitamin E and vitamin A), is easy to deteriorate when contacting with air, has similar nutritional ingredients to sunflower oil, is added into cosmetics generally used for moistening and lubricating skin, can improve eczema and rough skin, enables the skin to become tender and smooth, and helps to normalize metabolism. The meadowfoam seed oil contains more than 98% of long-chain fatty acid with an anti-oxidation effect, has a unique molecular structure, has a moisturizing function, can form a water-locking film on the surface of skin, locks water to resist dryness, can stretch skin texture, deeply repairs skin loss and improves the overall skin contour. The macadimia nut seed oil is characterized by being rich in unsaturated fatty acid, can prevent heart disease, can regulate and control blood sugar level, improves lipid metabolism of diabetics, can effectively moisturize and protect skin, and is a good sun-proof and skin-care product.

In some embodiments, the compositions of the present invention comprise 1 to 50% by weight of plant seed oil. In a preferred embodiment, the composition of the present invention comprises 40-50 wt% of vegetable seed oil.

In some embodiments, the compositions of the invention further comprise a plant squalane. The squalane is derived from olive oil, and is a natural skin caring oil. The squalane is a lipid closest to sebum of a human body, has strong affinity, can be integrated with the sebum membrane of the human body, forms a natural barrier on the surface of the skin, can inhibit the peroxidation of skin lipid, can effectively permeate into the skin, promotes the proliferation of skin basal cells, delays the skin aging, and simultaneously can open skin pores, promote blood circulation, promote cell metabolism and help repair damaged cells.

In some embodiments, the compositions of the present invention comprise 1 to 50% by weight plant squalane. In a preferred embodiment, the composition of the invention comprises from 40 to 50% by weight of plant squalane.

In some embodiments, the compositions of the present invention further comprise a deep sea fish oil. The deep sea fish oil is unsaturated fat components extracted from deep sea fish animals, namely EPA and DHA, has the effects of strengthening brain, benefiting intelligence, protecting cardiovascular system and guaranteeing smooth blood flow, and also has anti-inflammatory activity, so the deep sea fish oil is often used for improving various inflammations such as rheumatoid arthritis and the like.

In some embodiments, the compositions of the present invention comprise 1-50% by weight of the deep sea fish oil. In a preferred embodiment, the composition of the present invention comprises 30-40% by weight of deep sea fish oil.

In some embodiments, the compositions of the present invention further comprise a ceramide. In a specific embodiment, the ceramide is ceramide NP. Ceramide NP, also called ceramide 3, is a lipid substance that has a structure similar to that of the substances constituting the stratum corneum of the skin, and can rapidly permeate into the skin to combine with water in the stratum corneum to form a network structure to retain water. Has good effect on moisturizing and repairing skin, is an important skin-activating component in the horny layer, and can enhance the skin barrier and rebuild cells.

In some embodiments, the compositions of the present invention comprise 1-5% by weight ceramide. In a preferred embodiment, the composition of the invention comprises 2 to 3% by weight of ceramide.

In some embodiments, the fat composition of the present invention comprises shea butter, olive oil, safflower seed oil, meadowfoam seed oil, plant squalane, deep sea fish oil, ceramide 3, or combinations thereof.

The present invention has unexpectedly found that a composition comprising an oil or fat selected from any one or more of vegetable fruit oil (e.g., shea butter, olive oil, etc.), vegetable seed oil (e.g., safflower seed oil, meadowfoam seed oil, etc.), vegetable squalane, deep sea fish oil, ceramide 3 has skin barrier-related efficacy, can increase skin barrier activity and promote expression of skin barrier-related factors FLG, LOR, IVL, TGM1, CASP14, HMGCR, AQP3, and ZO-1.

In some embodiments, the compositions of the present invention comprise from 0.1 to 100% by weight of the oil.

In other embodiments, the compositions of the present invention comprise from 0.1 to 50 weight percent of the oil. In some embodiments, the compositions of the present invention comprise from 0.1 to 20 weight percent of a grease. In some embodiments, the compositions of the present invention comprise 0.5 to 10 weight percent of the oil.

In some embodiments of the invention, there is provided a composition comprising shea butter and plant squalane, wherein the weight ratio of shea butter to plant squalane is from 2:8 to 8: 2. In a preferred embodiment, the shea butter and squalane are present in a weight ratio of 1: 1.

Natural surfactant

The compositions of the present invention may also comprise natural surfactants. Natural surfactants are mostly derived from animal and plant bodies and are relatively complex high molecular organic substances. Because of its strong hydrophilicity, it can form an emulsion. These substances usually have a high viscosity, which is beneficial to emulsion stability.

Natural surfactants included in the compositions of the present invention include, but are not limited to: lecithin, cholesterol, lanolin, tea saponin, protein, saponins, saccharides, alkyl polyglycoside, etc. Lecithin exists in biological cells, such as animal egg, brain, etc., and plant seed or embryo, and yolk phospholipid is extracted from yolk; soybean is rich in lecithin. Lecithin has physiological activities of emulsification, dispersion, oxidation resistance and the like, and is a natural excellent surfactant and an important emulsifier.

The hydrogenated lecithin has strong hydrophilicity and moisture retention, has strong affinity to skin and mucous membrane, and can play the roles of moisture retention, emulsification, dispersion and the like when being used in a formula of cosmetics; as a surfactant, the oil-water-free emulsion can also condition the skin to achieve a good oil-water balance effect, and the hydrogenated lecithin can be used for developing advanced cosmetic products such as skin care cream, hand cream, lipstick, sunscreen oil and the like.

In some embodiments, the compositions of the present invention comprise from 0.1 to 20 wt% of a natural surfactant. In a preferred embodiment, the composition of the invention comprises from 0.1 to 10% by weight of natural surfactant. In a preferred embodiment, the composition of the invention comprises 0.1 to 5 wt% of a natural surfactant.

Drug and/or external preparation for skin

The oil and fat composition containing ceramide can be applied to medicines and/or skin external preparations for enhancing the skin barrier function.

In some embodiments, the pharmaceutical product is selected from: tablets, capsules, emulsions, suspensions, powders, granules, solutions, and various pharmaceutical dosage forms known in the art. Different amounts are added according to different types of dosage forms.

In some embodiments, the external skin agent is selected from: face cleaning lotion, cosmetic water, lotion, cream, jelly and facial mask. Different amounts are added according to different types of preparations.

The external preparation for skin is a general concept of all ingredients generally used for the external part of skin, and may be, for example, a cosmetic composition. The cosmetic composition can be basic cosmetics, face makeup cosmetics, body makeup cosmetics, hair care cosmetics and the like, and the dosage form of the cosmetic composition is not particularly limited and can be reasonably selected according to different purposes. The cosmetic composition also contains different cosmetically acceptable media or matrix excipients according to different dosage forms and purposes.

The external preparation for skin contains a dermatologically acceptable carrier or vehicle (e.g., lotion, cream, ointment, cleanser, etc.). One of ordinary skill in the art will be able to select a carrier that will dissolve or disperse the components in the concentrations described above, based on common general knowledge in the art.

One of ordinary skill in the art will be able to select suitable carriers, including for example water, alcohols, oils, and the like, based on the common general knowledge and their ability to dissolve or disperse in the active ingredient at the concentrations most suitable for treatment.

The skin external preparation of the present invention may be in the form of a topical application product, which can be externally applied to the skin, and can be prepared by those ordinary techniques well known in the art. The carrier may take any of a variety of practical forms such as a cream, dressing, gel, lotion, ointment or liquid comprising the applied and rinsed-off composition and incorporated into a carrier of material such as a dry or wet spread, hydrogel matrix, or adhesive (or non-adhesive) patch by methods well known in the art. Preferably, the carrier is a gel or a lotion that adds moisture, or a spread in dry or wet form.

Typical carriers include emulsions comprising water and/or an alcohol and an emollient such as hydrocarbon oils and waxes, silicone oils, hyaluronic acid, vegetable, animal or marine fats or oils, glyceride derivatives, fatty acids, or fatty acid esters or alcohols or alcohol ethers, lanolin and its derivatives, polyols or esters, wax esters, sterols, phospholipids and the like, and typically emulsifiers (nonionic, cationic or anionic), although some emollients inherently have emulsifying properties. In addition, these same components may be formulated into creams, gels, or solid sticks using different ratios of their components and/or by incorporating thickeners such as gums or other forms of hydrocolloids.

The skin external agent of the present invention may contain additional components commonly found in skin care compositions, such as emollients, skin conditioning agents, emulsifiers, preservatives, antioxidants, fragrances, chelating agents, and the like, as long as they are physically and chemically compatible with the other components of the skin external agent and do not affect the effect of the composition of the present invention.

In some embodiments of the external preparation for skin of the present invention, one or more preservatives may be used. Suitable preservatives include p-hydroxyacetophenone, C1-C4 alkyl parabens, and phenoxyethanol. Preservatives are used in amounts of about 0.5 to about 2 weight percent, preferably about 0.5 to 1 weight percent, based on the total weight of the composition.

In one example of the external preparation for skin of the present invention, one or more antioxidants may be used. Suitable antioxidants include Butylated Hydroxytoluene (BHT), ascorbyl palmitate (BHA), butylated hydroxyanisole, phenyl-alpha-naphthylamine, hydroquinone, gallopropyl, nordihydroguaiaretic acid, vitamin E or derivatives of vitamin E, vitamin C and its derivatives, calcium pantothenate, green tea extract and mixed polyphenols, and mixtures of the above. The antioxidant is used in an amount ranging from about 0.02 to 0.5 weight percent, more preferably from about 0.002 to 0.1 weight percent, based on the total weight of the composition.

In one example of the external preparation for skin of the present invention, one or more emollients may be used, functioning as a lubricant by their ability to remain on the surface of the skin or in the stratum corneum, to reduce flaking, and to improve the appearance of the skin. Typical emollients include fatty esters, fatty alcohols, mineral oils, polyether siloxane copolymers, and the like. Examples of suitable emollients include, without limitation, polypropylene glycol ("PPG") -15 stearyl ether, PPG-10 cetyl ether, Steareth-10, Oleth-8, PPG-4 lauryl ether, vitamin E acetate, lanolin, cetyl alcohol, cetearyl ethylhexanoate, cetearyl alcohol, glyceryl stearate, octyl hydroxystearate, dimethylpolysiloxane, and combinations thereof. Cetyl alcohol, cetearyl ethylhexanoate, cetostearyl alcohol, glyceryl stearate and combinations thereof are preferred. When used, the emollient is present in an amount ranging from about 0.1 to about 30 weight percent, preferably from about 1 to about 30 weight percent, based on the total weight of the composition.

In one example of the external preparation for skin of the present invention, one or more moisturizers may also be used. Humectants, also known as humectants, help to increase the effectiveness of the emollient, reduce flaking, stimulate the removal of compositional scales, and improve skin feel. Polyhydric alcohols may be used as humectants, including, but not limited to, glycerin, polyalkylene glycols, alkylene polyols and derivatives thereof, including butylene glycol, propylene glycol, dipropylene glycol, polyglycerol, polyethylene glycol and derivatives thereof, sorbitol, hydroxypropyl sorbitol, hexylene glycol, 1, 3-dibutylene glycol, 1,2, 6-hexanetriol, ethoxylated glycerin, propoxylated glycerin, and combinations thereof. When used, the humectant is present in an amount of about 0.1 to about 20 weight percent, preferably about 1 to about 15 weight percent, based on the total weight of the composition.

In one example of the external preparation for skin of the present invention, one or more emulsifiers may be used. The emulsifier may be used in an effective stabilizing amount range. Preferably, the emulsifier is used in an amount of about 1.0 to about 10.0 wt%, more preferably about 3.0 to about 6.0 wt%, based on the total weight of the composition. Any emulsifier that is compatible with the components of the composition may be used. Suitable emulsifiers include stearic acid, cetyl alcohol, glyceryl stearate, lecithin, stearyl alcohol, Steareth-2, Steareth-20, acrylic/C10-30 alkanol acrylate crosspolymer, and combinations thereof.

In one example of the external preparation for skin of the present invention, one or more pH adjusting agents may be used. The pH adjusting agent useful in the skin external preparation of the present invention includes tromethamine. When used, the pH adjusting agent is used in an amount of about 0.1 to about 2 weight percent, preferably about 0.1 to about 1 weight percent, based on the total weight of the composition.

In one embodiment of the present invention, the external preparation for skin comprises acrylic acid/C10-30 alkanol acrylate crosspolymer, glycerin, p-hydroxyacetophenone, glyceryl stearate and lecithin, cetostearyl alcohol, cetearyl ethyl hexanoate, tromethamine or a combination thereof.

In some embodiments of the present invention, the composition of the present invention is used in an amount of 0.001% to 100% (w/w), preferably 0.01% to 60% (w/w), and more preferably 0.01% to 40% (w/w) in the skin external preparation.

Examples

The invention will be further illustrated by the following specific examples. It should be noted that the examples are given solely for the purpose of illustration and are not to be construed as limitations on the scope of the invention, as many insubstantial modifications and variations of the invention may be made by those skilled in the art in light of the above teachings. Test methods in which specific conditions are not specified in the following examples are generally carried out under conventional conditions or under conditions recommended by the manufacturer. All percentages and parts are by weight unless otherwise indicated.

The experimental materials used in the following examples include:

butyrospermum PARKII (Butyrum PARKII) butter, a trade name of refined shea butter, available from basf New materials Co., Ltd

Olive (OLEA EUROPAEA) fruit oil, sold under the trade name refined olive oil, available from Textron tenicnica, s.l.u.

Safflower (CARTHAMUS TINCTORIUS) seed oil, commercially available as safflower seed oil, is available from Northstar Lipids UK Ltd.

Potentilla chinensis (LIMNANTHES ALBA) seed oil, tradename Potentilla chinensis seed oil, available from Haimines specialty Chemicals

Squalane, a tradename plant squalane, available from NIKKO CHEMICALS co.

Sea bream (Hoplostethous atlanticus) oil, which is a deep sea fish oil, was purchased from Nippon aquatic products Co., Ltd

Ceramide NP, tradename ceramide 3, available from Soxhlet materials GmbH

Hydrogenated lecithin, trade name hydrogenated lecithin, was purchased from NIKKO CHEMICALS co.

The experimental apparatus used in the examples included:

weighing a balance: PB4002-N type by METTLER TOLEDO;

a pH meter: SEVENMULTI model METTLER TOLEDO;

a water bath kettle: an electric heating constant temperature water bath kettle of HWS28 model of Shanghai constant science and technology Limited;

a homogenizer: POLYTRON PT 3100D;

a desk type stirrer: IKA @ EUROSTAR, power control-visc.

A magnetic stirrer:

C-MAG HP 7。

example 1: preparation of oil and fat composition

Weighing 0.5 part by mass of hydrogenated lecithin in a beaker, adding 99.5 parts by mass of deionized water, and uniformly stirring for later use.

Example 2: preparation of oil and fat composition

Weighing 100 parts by mass of plant squalane to be tested.

Example 3: preparation of oil and fat composition

Weighing 100 parts by mass of shea butter and testing.

Example 4: preparation of oil and fat composition

Weighing 0.5 part by mass of hydrogenated lecithin and 5 parts by mass of shea butter in a beaker, heating to 75-80 ℃ while stirring, then weighing 94.5 parts by mass of deionized water at 75-80 ℃ in the beaker, starting homogenizing at 3000 r/min for 5-10 min for later use.

Example 5: preparation of oil and fat composition

Weighing 0.5 part by mass of hydrogenated lecithin, 5 parts by mass of shea butter and 5 parts by mass of olive oil in a beaker, heating to 75-80 ℃ while stirring, then weighing 89.5 parts by mass of deionized water at 75-80 ℃ in the beaker, starting homogenizing at 3000 r/min for 5-10 min for later use.

Example 6: preparation of oil and fat composition

Weighing 0.5 part by mass of hydrogenated lecithin, 5 parts by mass of shea butter and 5 parts by mass of safflower seed oil, heating to 75-80 ℃ while stirring, then weighing 89.5 parts by mass of deionized water at 75-80 ℃ in a beaker, starting homogenizing at 3000 r/min for 5-10 min for later use.

Example 7: preparation of oil and fat composition

Weighing 0.5 part by mass of hydrogenated lecithin, 5 parts by mass of shea butter and 5 parts by mass of white pond flower seed oil in a beaker, heating to 75-80 ℃ while stirring, then weighing 89.5 parts by mass of deionized water at 75-80 ℃ in the beaker, starting homogenizing at 3000 r/min for 5-10 min for later use.

Example 8: preparation of oil and fat composition

Weighing 0.5 part by mass of hydrogenated lecithin, 5 parts by mass of shea butter and 5 parts by mass of plant squalane in a beaker, heating to 75-80 ℃ while stirring, then weighing 89.5 parts by mass of deionized water at 75-80 ℃ in the beaker, starting homogenizing at 3000 r/min for 5-10 minutes for later use.

Example 9: preparation of oil and fat composition

Weighing 4.76 parts by mass of hydrogenated lecithin, 47.62 parts by mass of shea butter and 47.62 parts by mass of plant squalane in a beaker, heating to 75-80 ℃ while stirring, completely dissolving uniformly, and cooling to room temperature for later use.

Example 10: preparation of oil and fat composition

Weighing 3.22 parts by mass of hydrogenated lecithin, 32.26 parts by mass of shea butter, 32.26 parts by mass of plant squalane and 32.26 parts by mass of deep sea fish oil in a beaker, heating to 75-80 ℃ while stirring, completely dissolving uniformly, and cooling to room temperature for later use.

Example 11: preparation of oil and fat composition

Weighing 3.22 parts by mass of hydrogenated lecithin, 32.26 parts by mass of shea butter, 32.26 parts by mass of plant squalane and 32.26 parts by mass of white pond flower seed oil in a beaker, heating to 75-80 ℃ while stirring, completely dissolving uniformly, and cooling to room temperature for later use.

Example 12: preparation of oil and fat composition

Weighing 3.22 parts by mass of hydrogenated lecithin, 32.26 parts by mass of shea butter, 32.26 parts by mass of plant squalane and 32.26 parts by mass of safflower seed oil, heating to 75-80 ℃ while stirring, completely dissolving uniformly, and cooling to room temperature for later use.

Example 13: preparation of oil and fat composition

Weighing 4.63 parts by mass of hydrogenated lecithin, 46.30 parts by mass of shea butter, 46.30 parts by mass of vegetable squalane and 2.77 parts by mass of ceramide 3 in a beaker, heating to 75-80 ℃ while stirring, completely dissolving uniformly, and cooling to room temperature for later use.

Test example 1: 3D epidermal recombination model tissue viability test

The 3D epidermal recombination model is a human 3D epidermal model and can simulate the structure of each layer of the epidermis differentiated by keratinocytes. According to the 3D epidermal recombination model tissue viability testing method, firstly, a certain damage is caused to the model by using a skin irritant SDS, and then, a proper amount of sample prepared by the embodiment is covered on the surface of the damaged model for 24 hours to repair. After 24 hours, the samples were washed away and tested for tissue viability recovery using the MTT method. The skin barrier repair ability of the example samples can be evaluated according to the tissue viability recovery.

1. Test materials

1.1 cells: the skin model is a 3D epidermal skin model

(EpiSkin)。

1.2 reagent:

3D skin model maintenance medium, 3D skin model detection medium (EpiSkin); sodium Dodecyl Sulfate (SDS) (Sigma); calcium and magnesium containing dpbs (gibco); mtt (sigma); isopropanol and concentrated hydrochloric acid (national drug group).

1.3 Main Equipment

CO₂An incubator HERAcell 240i, a biological safety cabinet 1300SERIES A2, a microplate reader Multiskan Mk3 (Thermo); forward pipettor M25 (Gilson); 50mL Multipette plus (Eppendorf) continuous applicator tip; skin punch (area 0.38 cm)²)。

2. The experimental method comprises the following steps:

3D recombinant epidermal model reception and resuscitation

Receiving the 3D epidermal recombination model, checking the integrity of the package, checking the number of models, checking the maintenance medium, and detecting the color and volume of the medium. And after the error is confirmed, starting to perform model recovery.

The resuscitation operation is performed in a biosafety cabinet. A number of 12-well plates were taken, 2 ml/well of maintenance medium was added to the first row, then the 3D recombinant epidermis model was removed with forceps, the surface gel was carefully removed with a cotton swab, placed in the wells containing maintenance medium, and allowed to resuscitate for 24 hours. One 12-well plate can resuscitate 4 3D recombinant epidermal models. The culture conditions are as follows: 5% CO₂And culturing in a constant-temperature incubator at 37 ℃.

3D recombination epidermis model damage treatment and sample adding product

A1.5% (w/v) Sodium Dodecyl Sulfate (SDS) solution was prepared using a 1 XPhosphate buffer solution (DPBS).

The model was divided into a blank control group (3 replicates without any treatment), a negative control group (3 replicates with 1.5% SDS) and sample groups of examples 1-13 (3 replicates per sample).

2 ml/well of maintenance medium was added to the second row of the 12-well plate, and then the 3D recombinant epidermal model located in the first row was transferred to the corresponding second row with tweezers.

50 microliters of 1.5% SDS solution was dropped on the surfaces of the negative control group and the sample group models of examples 1 to 6 and examples 8 to 9 using a pipette, andensure uniform coverage and incubate for 15 min at room temperature. After 15 minutes, immediately wash with 1 × DPBS and gently wipe the residual liquid with a cotton swab. Then, 15. mu.l of the sample was applied to the surface of the lesion-treated (1.5% SDS) sample group model by using a piston displacement pipette (M25) and treated for 24 hours. All models were placed in a cell incubator under the following culture conditions: 5% CO₂And culturing in a constant-temperature incubator at 37 ℃.

3D recombinant epidermal model viability test

3 mg/ml of thiazole blue (MTT) stock was prepared with 1 XDPBS and the MTT stock was diluted to 0.3 mg/ml in the test medium. 2 ml/well of diluted MTT solution was added in the third row of the 12-well plate.

The sample on the surface of the model was washed off (not directly aligned for washing, but aligned to the side wall of the skin model support frame) by pipetting 1 XDPBS with a 50ml continuous pipette, and then pipetting the residual liquid with a cotton swab into a 12-well plate in the third line MTT solution. Finally placing the mixture into a cell culture box, and placing the cell culture box at 37 ℃ and 5% CO₂Was incubated in the incubator of (1) for 3 hours.

The acidic isopropanol was prepared and 1.8 ml of hydrochloric acid was added to 500 ml of isopropanol. A corresponding number of 1.5 ml microcentrifuge tubes (EP tubes) were molded and 500. mu.l of the acidic isopropanol was added to each tube. After the epidermal tissue was incubated in MTT for 3 hours, the epidermal tissue was removed with a punch, separated from the underlying collagen matrix with two small-gauge elbow forceps, and inverted, and then transferred together with the collagen matrix to a 1.5 ml EP tube containing acidic isopropanol. The EP tube was shaken and mixed on a vortex, and placed in a refrigerator at 4 ℃ for 72 hours for color extraction. Meanwhile, prepare an EP tube containing 1 ml of acid isopropanol, as the blank hole when the enzyme-linked immunosorbent assay detects.

The 1.5 ml EP tube containing the acid isopropanol and the skin model was taken out from the 4 ℃ freezer, the solution was mixed by shaking, 150 μ l of the extraction solution was taken out per tube and transferred to a 96-well plate, and 3 times of the extraction solution were taken out and added to the 96-well plate. Meanwhile, 3-6 wells of acidic isopropanol were prepared as blank control during detection, excluding background optical density (OD value). The OD was read at a wavelength of 570 nm. Relative activity of each group of models (blank control 100%) was calculated according to the following formula:

3. result judgment

The blank control group is an undamaged group, and the relative activity is 100%; the relative activity of the negative control group is controlled to be 40-60% compared with that of the blank control group; a higher relative activity in the sample group than in the negative control group indicates that the sample group has the ability to promote barrier repair, and a lower relative activity in the sample group than in the negative control group indicates that the sample group does not have the ability to promote barrier repair.

Table 1: MTT method for detecting relative activity condition of each group of tissues

As shown in table 1, the average relative viability of the skin model tissue of the blank control group was 100%, and the average relative viability of the skin model tissue of the negative control group (1.5% SDS treatment) was 41.0%. Example 1 the average relative skin model tissue viability was 48.4%, example 2 the average relative skin model tissue viability was 65.8%, example 3 the average relative skin model tissue viability was 70.8% (100% shea butter), example 4 the average relative skin model tissue viability was 75.3% (5% shea butter), example 5 the average relative skin model tissue viability was 69.0% (5% shea butter + 5% olive oil), example 6 the average relative skin model tissue viability was 54.3% (5% shea butter + 5% safflower seed oil), example 7 the average relative skin model tissue viability was 68.0% (5% shea butter + 5% chinchilla seed oil), example 8 the average relative skin model tissue viability was 57.0%, example 9 (0.5% hydrogenated lecithin + 5% shea butter + 5% vegetable squalane 5%) skin model tissue average relative viability was 89.9%, example 10 (0.5% hydrogenated lecithin + 5% shea butter + 5% vegetable squalane + 5% deep sea fish oil) skin model tissue average relative viability was 39.9%, example 11 (0.5% hydrogenated lecithin + 5% shea butter + 5% vegetable squalane + 5% margaria japonica seed oil) skin model tissue average relative viability was 85.9%, example 12 (0.5% hydrogenated lecithin + 5% shea butter + 5% vegetable squalane + 2%) skin model tissue average relative viability was 84.1%, example 13 (0.5% hydrogenated lecithin + 5% shea butter + 5% vegetable squalane + 30.3%) skin model tissue average relative viability was 91.3% (see table 1).

In conclusion, the tissue repair capacity of the samples of example 9 was higher than that of the samples of examples 2 to 4, and also higher than that of the samples of examples 6 to 8. In addition, the tissue repair ability of the sample of example 9 was higher than that of examples 10 to 12 and lower than that of the sample of example 13.

Test example 2: 3D recombinant skin epidermal model barrier-related gene expression test

The 3D epidermal recombination model is a human 3D epidermal model and can simulate the structure of each layer of the epidermis differentiated by keratinocytes. Covering a proper amount of samples on an epidermis model for 24 hours, then collecting total ribonucleic acid (RNA) of epidermis tissues, and quantitatively detecting related genes of the skin barrier by adopting an RNA reverse transcription technology and a gene microarray technology so as to evaluate the skin barrier moisturizing and repairing capacity of the samples.

1. Test materials

1.1 cells: the skin model is a 3D epidermal skin model

(EpiSkin)。

1.2 reagent:

3D skin model maintenance medium (EpiSkin); calcium and magnesium containing dpbs (gibco); isopropanol, 75% absolute ethanol (national pharmaceutical group); DEPC water (tiangen organisms); total RNA extraction reagent trizol (thermofisher); reverse transcription Kit iScript cDNA Synthesis Kit and fluorescent real-time quantitative PCR reagent iTaq^TM Universal

Green Supermix(Bio-rad)。

1.3 Main Equipment

CO₂Incubator HERACell 240i, biosafety cabinet 1300SERIES A2, ultramicro UV spectrophotometer NanoDrop ONE (Thermo); gene amplification instrument C1000 Touch, real-time quantitative PCR instrument CFX Connect (Bio-rad); high speed refrigerated centrifuge MIKRO 220r (hittich); forward pipettor M25 (Gilson); 50mL Multipette plus (Eppendorf) continuous applicator tip; skin punch (area 0.38 cm)²)。

2. The experimental method comprises the following steps:

3D recombinant epidermal model reception and resuscitation

3D recombinant epidermal model sample treatment

The model was divided into the sample groups of examples 1, 4-8 (3 replicates per sample) and the positive control group (rosiglitazone, 3 replicates).

A piston displacement pipettor (M25) was used to add 15 microliters of rosiglitazone solution (100 micromoles in 1 x DPBS) to the positive control group, 15 microliters of sample was added to the sample group, and the samples were gently spread evenly over the skin model surface using a suction nozzle, in triplicate for each sample. All models were placed in a cell incubator and incubated for 24 hours. The culture conditions are as follows: 5% CO₂And culturing in a constant-temperature incubator at 37 ℃.

3D recombinant epidermal model Collection of Total RNA samples

The 1 XDPBS was aspirated with a 50ml continuous applicator, the sample was rinsed off the surface of the model (the sample was not directly aligned for rinsing, but was aligned with the side wall of the skin model support frame), and then the residual liquid was aspirated with a cotton swab. The epidermal tissue was cut with a punch, separated from the underlying collagen base with two small-gauge bent tweezers, placed in an EP tube containing 1 ml of total RNA extraction reagent (TRIzol), and repeatedly tapped with 1 ml of a rnase-free pipette tip until only the transparent stratum corneum remained. Subsequently, 0.2 ml of chloroform was added to the original EP tube, vigorously shaken, and centrifuged at a high speed at a low temperature for 15 minutes. Taking 500 microliters of the upper aqueous phase by using a pipette nozzle without RNase, adding equal volume of precooled (minus 20 ℃) isopropanol, reversing, mixing uniformly, and centrifuging at a high speed for 10 minutes at a low temperature. The centrifuged RNA forms a white flaky precipitate at the bottom of the EP tube, the precipitate is rinsed twice with 75% ethanol, residual ethanol is removed by a centrifuge and a suction nozzle, and the precipitate is opened and dried to be transparent at room temperature. With 40. mu.l RNase-free H₂Dissolving O, and mixing. The RNA concentration was measured again using a microspectrophotometer Nanodrop and the integrity of the RNA was checked by agarose gel electrophoresis. The dissolved total RNA of the epidermal model tissue is used for subsequent experiments or is stored for a long time at minus 80 ℃ after being subpackaged.

Quantitative detection of skin barrier related gene by RT-qPCR method

Taking 1 microgram of total RNA for reverse transcription, and adopting a reverse transcription Kit iScript cDNA Synthesis Kit for reverse transcription to obtain cDNA.

The reverse transcription system is as follows:

the reverse transcription procedure was as follows: 5 minutes at 25 ℃; 30 minutes at 42 ℃; 5 minutes at 85 ℃.

Real-time quantification of Polymerase Chain Reaction (PCR) reagent iTaq using fluorescence^TM Universal

Green Supermix is used for carrying out fluorescence real-time quantitative detection, and the expression condition of messenger RNA (mRNA) of the barrier related gene is analyzed.

The reaction system is as follows:

the reaction procedure was as follows: at 95 ℃ for 30 seconds; at 95 deg.C, 15 seconds, 60 deg.C, 30 seconds, 40 cycles; 65-95 deg.C, each time by 0.5 deg.C (melting curve).

The primer sequences used in this example were designed as follows:

by using

The gene expression of the sample group and the positive control group (rosiglitazone) of example was analyzed for changes relative to the gene expression of the control group of example 1.

Result judgment

Example 1 the gene expression levels of the sample groups were taken as 100%, and each group compared to it, more than 100%, indicated up-regulation of gene expression, and less than 100%, indicated down-regulation of gene expression. FLG, LOR and other genes are up-regulated, which indicates that the sample group has the capacity of moisturizing and promoting barrier repair; genes such as FLG, LOR were down-regulated, indicating that this sample group did not have the ability to moisturize and promote barrier repair.

Table 2: skin barrier factor expression after treatment of each group of samples

As shown in Table 2, the relative gene expression level of example 1 was 100% as a control, and the gene expression levels of the samples of examples 4, 5, 6, 7, 8, 9, 10, 11, 12, and 13 and the positive control sample were compared to obtain the corresponding percentages.

Specifically, the barrier factor FLG was found in the samples of example 4 (shea butter 5%), the samples of example 5 (shea butter 5% + olive oil 5%), the samples of example 6 (shea butter 5% + safflower seed oil 5%), the samples of example 7 (shea butter 5% + chinchilla seed oil 5%), the samples of example 8 (shea butter 5% + vegetable squalane 5%), the samples of example 9 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5%), the samples of example 10 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5% + deep sea fish oil 5%), the samples of example 11 (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + chinchilla seed oil 5%), the samples of example 12 (hydrogenated lecithin 0.5% + squalane 5% + vegetable squalane 5% + safflower seed oil 2%), and the samples of example 5% + chinchilla seed oil 5% + safflower seed oil 2%, The samples of example 13 (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + ceramide III 0.3%) and the positive control (rosiglitazone) were treated with 222%, 265%, 283%, 223%, 186%, 305%, 147%, 274%, 253%, 247% and 362% respectively, compared to the samples of example 1.

Specifically, the barrier factor LOR is calculated for the samples of example 4 (shea butter 5%), the samples of example 5 (shea butter 5% + olive oil 5%), the samples of example 6 (shea butter 5% + safflower seed oil 5%), the samples of example 7 (shea butter 5% + chinchilla seed oil 5%), the samples of example 8 (shea butter 5% + vegetable squalane 5%), the samples of example 9 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5%), the samples of example 10 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5% + deep sea fish oil 5%), the samples of example 11 (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + vegetable squalane 5% + chinchilla seed oil 5%), the samples of example 12 (hydrogenated lecithin 0.5% + squalane vegetable squalane 5% + safflower seed oil 2%) + safflower seed oil 2% + olive oil 5% and the samples 5% in the like, The samples of example 13 (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + ceramide III 0.3%) and the positive control (rosiglitazone) were treated with 240%, 242%, 247%, 208%, 190%, 239%, 136%, 218%, 244%, 236% and 351%, respectively, compared to the samples of example 1.

Specifically, the barrier factor IVL was found in the samples of example 4 (shea butter 5%), the samples of example 5 (shea butter 5% + olive oil 5%), the samples of example 6 (shea butter 5% + safflower seed oil 5%), the samples of example 7 (shea butter 5% + chinchilla seed oil 5%), the samples of example 8 (shea butter 5% + vegetable squalane 5%), the samples of example 9 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5%), the samples of example 10 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5% + deep sea fish oil 5%), the samples of example 11 (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + chinchilla seed oil 5%), the samples of example 12 (hydrogenated lecithin 0.5% + squalane 5% + vegetable squalane 5% + safflower seed oil 2%) + safflower seed oil 2% + olive oil 5% + squalane seed oil 5% + squalane seed oil 5% + squalane seed oil 5% + squalane oil 5% and the samples of example 12, After treatment of the sample of example 13 (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + ceramide III 0.3%) and the positive control (rosiglitazone), 113%, 107%, 109%, 125%, 123%, 136%, 119%, 103%, 101%, 114% and 103%, respectively, compared with the sample of example 1.

Specifically, the barrier factor TGM1 was found in the samples of example 4 (shea butter 5%), example 5 (shea butter 5% + olive oil 5%), example 6 (shea butter 5% + safflower seed oil 5%), example 7 (shea butter 5% + chinquai seed oil 5%), example 8 (shea butter 5% + vegetable squalane 5%), example 9 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5%), example 10 (hydrogenated lecithin 0.5% + squalane 5% + fish oil 5%), example 11 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5% + fish oil 5%), example 12 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5% + safflower seed oil 5%), and example 12 (hydrogenated lecithin 0.5% + squalane vegetable squalane 5% + safflower seed oil 2%) + safflower seed oil 5% +, The samples of example 13 (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + ceramide III 0.3%) and the positive control (rosiglitazone) were treated with 134%, 170%, 176%, 201%, 165%, 108%, 161%, 183%, 224% and 161%, respectively, compared to the samples of example 1.

Specifically, the barrier factor CASP14 was found in the samples of example 4 (shea butter 5%), example 5 (shea butter 5% + olive oil 5%), example 6 (shea butter 5% + safflower seed oil 5%), example 7 (shea butter 5% + chinquai seed oil 5%), example 8 (shea butter 5% + vegetable squalane 5%), example 9 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5%), example 10 (hydrogenated lecithin 0.5% + squalane butter 5% + vegetable squalane 5% + fish oil 5%), example 11 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5% + fish oil 5%), example 12 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5% + safflower seed oil 5%), and example 12 (hydrogenated lecithin 0.5% + vegetable squalane 5% + safflower seed oil 2%) + safflower seed oil 5% +, The samples of example 13 (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + ceramide III 0.3%) and the positive control (rosiglitazone) were treated with 188%, 201%, 228%, 162%, 142%, 190%, 121%, 161%, 150%, 155% and 330% respectively, compared to the samples of example 1.

Specifically, the barrier factor HMGCR was found in the samples of example 4 (shea butter 5%), the samples of example 5 (shea butter 5% + olive oil 5%), the samples of example 6 (shea butter 5% + safflower seed oil 5%), the samples of example 7 (shea butter 5% + chinchilla seed oil 5%), the samples of example 8 (shea butter 5% + vegetable squalane 5%), the samples of example 9 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5%), the samples of example 10 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5% + fish oil 5%), the samples of example 11 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5% + chinzan seed oil 5%), the samples of example 12 (hydrogenated lecithin 0.5% + vegetable squalane 5% + safflower seed oil 2%) + squalane 5% + squalane, The samples of example 13 (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + ceramide III 0.3%) and the positive control (rosiglitazone) were treated with 170%, 208%, 253%, 213%, 170%, 205%, 211%, 142%, 143%, 185% and 229% respectively compared to the samples of example 1.

Specifically, the barrier factor AQP3 was found in the samples of example 4 (shea butter 5%), the samples of example 5 (shea butter 5% + olive oil 5%), the samples of example 6 (shea butter 5% + safflower seed oil 5%), the samples of example 7 (shea butter 5% + chinquai seed oil 5%), the samples of example 8 (shea butter 5% + vegetable squalane 5%), the samples of example 9 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5%), the samples of example 10 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5% + fish oil 5%), the samples of example 11 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5% + chini seed oil 5%), the samples of example 12 (hydrogenated lecithin 0.5% + vegetable squalane 5% + safflower seed oil 2%) + safflower seed oil 5% +, The samples of example 13 (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + ceramide III 0.3%) and the positive control (rosiglitazone) were treated with 112%, 138%, 147%, 128%, 130%, 126%, 148%, 132%, 136% and 131% respectively, compared to the samples of example 1.

Specifically, barrier factor ZO-1 was determined for the samples of example 4 (shea butter 5%), the samples of example 5 (shea butter 5% + olive oil 5%), the samples of example 6 (shea butter 5% + safflower seed oil 5%), the samples of example 7 (shea butter 5% + chinchilla seed oil 5%), the samples of example 8 (shea butter 5% + vegetable squalane 5%), the samples of example 9 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5%), the samples of example 10 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5% + fish oil 5%), the samples of example 11 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5% + chinella seed oil 5%), and the samples of example 12 (hydrogenated lecithin 0.5% + shea butter 5% + vegetable squalane 5% + safflower seed oil 2%) + safflower seed oil 5% +, The samples of example 13 (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + ceramide III 0.3%) and the positive control (rosiglitazone) were treated with 143%, 229%, 237%, 215%, 198%, 183%, 164%, 161%, 152%, 167% and 246% respectively, compared to the samples of example 1.

In conclusion, the sample of example 4 (shea butter 5%) promoted the expression of FLG, LOR, IVL, TGM1, CASP14, HMGCR, AQP3 and ZO-1 barrier factors, facilitating barrier formation, and had skin moisturizing and barrier repair promoting effects. Example 5 samples (shea butter 5% + olive oil 5%) promoted the expression of FLG, LOR, IVL, TGM1, CASP14, HMGCR, AQP3 and ZO-1 barrier factors, favouring barrier formation, with skin moisturizing and barrier repair promoting efficacy. Example 6 samples (shea butter 5% + safflower seed oil 5%) promoted the expression of FLG, LOR, IVL, TGM1, CASP14, HMGCR, AQP3 and ZO-1 barrier factors, favouring barrier formation, with skin moisturizing and barrier repair promoting efficacy. Example 7 samples (shea butter 5% + meadowfoam seed oil 5%) promoted the expression of FLG, LOR, IVL, TGM1, CASP14, HMGCR and ZO-1 barrier factors, favoured barrier formation, and had skin moisturizing and barrier repair promoting efficacy. Example 8 samples (Shea butter 5% + squalane 5%) promoted the expression of FLG, LOR, IVL, TGM1, CASP14, HMGCR, AQP3 and ZO-1 barrier factors, favouring barrier formation and having skin moisturizing and barrier repair promoting efficacy. Example 9 samples (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5%) promoted expression of FLG, LOR, IVL, TGM1, CASP14, HMGCR, AQP3 and ZO-1 barrier factors, favoured barrier formation, and had skin moisturizing and barrier repair promoting efficacy. Example 10 samples (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + deep sea fish oil 5%) promoted expression of FLG, LOR, IVL, TGM1, CASP14, HMGCR, AQP3 and ZO-1 barrier factors, favoured barrier formation and had skin moisturizing and barrier repair promoting efficacy. Example 11 samples (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + meadowfoam seed oil 5%) promoted the expression of FLG, LOR, IVL, TGM1, CASP14, HMGCR, AQP3 and ZO-1 barrier factors, favoring barrier formation, with skin moisturizing and barrier repair efficacy. Example 12 samples (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + safflower seed oil 2%) promoted the expression of FLG, LOR, IVL, TGM1, CASP14, HMGCR, AQP3 and ZO-1 barrier factors, favoured barrier formation, and had skin moisturizing and barrier repair benefits. Example 13 samples (hydrogenated lecithin 0.5% + shea butter 5% + squalane 5% + ceramide III 0.3%) promoted the expression of FLG, LOR, IVL, TGM1, CASP14, HMGCR, AQP3 and ZO-1 barrier factors, favouring barrier formation, with skin moisturizing and barrier repair benefits.

Application example

The oil and fat composition containing ceramide can be used as an intermediate raw material or a functional additive for preparing medicines or skin external preparations, and the skin external preparations are preferably cosmetic compositions, including but not limited to preparation of products in dosage forms of cream, lotion, gel, lotion, essence, facial mask, eye cream, aerosol (cleaning foam), spray, shower gel, massage oil, facial cleanser and the like.

The weight percentage of the oil and fat composition containing ceramide in the skin external preparation is 0.001-100% (w/w); preferably 0.01-60% (w/w); more preferably 0.01-40% (w/w).

The following are specific examples of the application of the compositions obtained in examples 4, 6, 7, 8 and 13 to external preparations for skin, and the formulation and preparation methods of these preparations. The specific application examples are as follows:

application example 1: preparation of face cream

Application example 2: preparation of the emulsion

Application example 3: preparation of eye cream

Application example 4: preparation of facial mask

Application example 5: preparation of essence

Application example 6: examples 2 to 13 in the present invention can also be used alone as a skin external preparation alone.

Claims

1. An oil composition comprising ceramide, wherein the oil is selected from the group consisting of: vegetable oil and fat, animal oil and fat or their combination,

wherein the vegetable oil is selected from: shea butter, olive oil, safflower seed oil, meadowfoam seed oil, plant squalane, or combinations thereof,

wherein the animal fat is deep sea fish oil,

wherein the composition comprises 0.01 to 5 wt% ceramide, based on the total weight of the composition.

2. The composition of claim 1, further comprising a natural surfactant.

3. The composition of claim 2, wherein the natural surfactant is selected from the group consisting of: lecithin, cholesterol, lanolin, tea saponin, protein, saponins, saccharides, alkyl polyglycoside or their combination.

4. The composition of claim 2, wherein the natural surfactant is hydrogenated lecithin.

5. The composition of any of claims 2-4, wherein the weight ratio of the oil to the natural surfactant is from 1:1 to 30: 1.

6. The composition of claim 5, wherein the weight ratio of the oil to the natural surfactant is from 10:1 to 30: 1.

7. Use of a composition according to claim 1 for enhancing the barrier function of skin.

8. Use according to claim 7, wherein the enhancement of skin barrier function is achieved by increasing skin barrier activity and/or promoting expression of skin barrier related factors.

9. Use of the composition according to claim 1 for the preparation of a medicament and/or a skin external preparation for enhancing the barrier function of the skin.

10. The use according to claim 9, wherein the pharmaceutical and/or dermatological preparations for external use comprise 0.001 to 100% by weight of the composition.