CN115607510B

CN115607510B - High-concentration oleanolic acid nanoemulsion composition and preparation method thereof

Info

Publication number: CN115607510B
Application number: CN202110419745.1A
Authority: CN
Inventors: 林振放
Original assignee: Quanzhou Daxun Biotechnology Co ltd
Current assignee: Quanzhou Daxun Biotechnology Co ltd
Priority date: 2021-04-19
Filing date: 2021-04-19
Publication date: 2024-03-08
Anticipated expiration: 2041-04-19
Also published as: CN115607510A

Abstract

A high-concentration oleanolic acid nanoemulsion composition and a preparation method thereof belong to the technical fields of medicines and cosmetics. The oleanolic acid nanoemulsion composition comprises 0.01% -8% of oleanolic acid; 1% -40% of a surfactant; 1% -40% of cosurfactant; terpene-containing essential oil 0.2% -20%; 0% -2% of baicalin; 0-5% of oleuropein; a proper amount of additive; the balance being water. Adding cosurfactant and terpene-containing essential oil into a container A, and uniformly mixing and stirring; adding a surfactant, and uniformly mixing; then adding oleanolic acid and other active ingredients, and stirring until the oleanolic acid and other active ingredients are completely dissolved; adding water and additives into a container B to obtain a water phase; slowly adding the mixture of the container A into the water phase of the container B while stirring to obtain primary emulsion; high-speed shearing and emulsifying, and secondary emulsifying. Can be used in preparing cosmetics, and can be used in preparing medicines for treating skin diseases, and can be used for caring skin, hair, etc.

Description

High-concentration oleanolic acid nanoemulsion composition and preparation method thereof

Technical Field

The invention belongs to the technical field of medicines and cosmetics, and particularly relates to a high-concentration oleanolic acid nanoemulsion composition and a preparation method thereof.

Background

Oleanolic acid (Oleanolic acid), also known as angelic acid, is a natural pentacyclic triterpene compound that is found in a wide variety of plants, both as an episome and as a glycoside. The content is generally 0.2% -2%. The powder purified by oleanolic acid is white crystal, odorless and tasteless. Is unstable to acid and alkali. Melting point 308-310 ℃, [ alpha ]20D+73.3° (c=0.15, chloroform), insoluble in water.

Oleanolic acid has wide pharmacological effects such as antibacterial, antiinflammatory, antioxidant, antitumor, liver protecting, antidiabetic, hair growth promoting, wound healing promoting, scar formation resisting, collagen formation promoting, and antiallergic effects. At present, oleanolic acid is clinically used for the adjuvant therapy of acute and chronic hepatitis, and is safe and has no toxic or side effect. The development and utilization of oleanolic acid except liver protection are not obvious, and the clinical preparation of the oleanolic acid mainly comprises two types of tablets and capsules.

Notably, some oleanolic acid-rich herbal plants, such as the traditional Chinese medicine ligustrum lucidum, are traditionally used in china for the treatment of hair loss, promotion of hair regrowth and hair blacking; oleanolic acid has been shown to up-regulate Wnt/β -catenin signaling pathway to promote hair growth and prolong the anagen phase of hair follicles in vitro (Liu B, et al, β -Catenin is involved in oleanolic acid-dependent promotion of proliferation in human hair matrix cells in an in vitro organ culture model [ J ]. Fitterapia, 2017, 121:136-140.); oleanolic acid has been shown to promote proliferation of human hair follicle stem cells cultured in vitro (Liu Ben, chen Xianyan, yi Huan, etc. effects of oleanolic acid on proliferation of human hair follicle stem cells cultured in vitro [ J ]. J. Skin disease, chem., 2017,24 (2): 68-71.); oleanolic acid has been shown to down-regulate gene expression of 5A REDUCTASE and androgen receptor in cultured hair follicles in vitro, and up-regulate expression levels of insulin-like growth factor (IGF-1), vascular Endothelial Growth Factor (VEGF) (CaoC, liu Y, et al EFFECT OF OLEANOLIC ACID ON A-REDUCTASE ACTITY, DPCS PROLIFERATION AND GENE EXPRESSION CORRELATED WITH ANDROGENETIC ALOPECIA IN VITRO [ J ]. ACTA MEDICA MEDITERRANEA,2019,35 (4): 2159-2165.); these studies show that oleanolic acid has a broad application prospect as a product for promoting hair growth and reducing hair loss.

The oleanolic acid has good supplementing or replacing effect on the existing hair growth or hair loss treatment; the U.S. Food and Drug Administration (FDA) has approved only two drugs for the treatment of male androgenetic alopecia (AGA): topical minoxidil (minoxidil) and oral finasteride (finasteride); finasteride is an inhibitor of 5 alpha-reductase type 2, which inhibits DHT-induced hair follicle miniaturization by inhibiting the conversion of testosterone hormone (T) to Dihydrotestosterone (DHT), resulting in reduced DHT formation; finasteride is generally more effective than minoxidil for AGA patients; however, finasteride has the potential to cause male sexual side effects and cause anxiety and depression; minoxidil is a nonspecific hair growth promoting agent whose mechanism of promoting hair growth is not completely known, and minoxidil is a potassium channel opener which can promote hair growth by dilating blood vessels and allowing more blood and nutrients to enter hair follicles; common adverse effects of minoxidil are mild dermatitis, itching, redness or irritation of the scalp and hirsutism, even exacerbation of hair loss, and the "Rogaine" manufacturer indicates that minoxidil causes hair loss to be a common side effect and describes the process as "shed phase"; and the minoxidil is easy to drop to the level before the minoxidil is used after stopping the drug; minoxidil can inhibit collagen production, possibly accelerating skin aging; serious allergic reactions may occur including rash, chest distress, chest pain, dizziness, tachycardia, etc.; existing therapeutic agents are not very successful and often have undesirable side effects that deeply plague patients and dermatologists; therefore, the development of effective oleanolic acid external preparations is particularly important.

Oleanolic acid is hardly soluble in water (solubility in water: 4.61mg/L at 25 ℃) and has poor cell permeability, and oleanolic acid belongs to class VI drugs, i.e., low-solubility, low-permeability drugs, according to the biopharmaceutical classification system. And the oleanolic acid has poor solubility and low dissolution efficiency in common alcohols, fatty acids, surfactants and other fat-soluble solvents, thereby greatly limiting the application of the oleanolic acid. The choice of a suitable formulation or solvent is critical to enhance the solubility of oleanolic acid, as only the dissolved drug can penetrate the skin.

The disclosed patent on oleanolic acid preparation mentions Gao Jidun that the dispersibility of the oleanolic acid in water, but still has problems that prevent large-scale industrial production and application, such as low oleanolic acid content, and complex organic solvents or processing steps involved in the damage to human skin or health.

For example, chinese patent CN101991532B discloses a self-microemulsion composition without water, said self-microemulsion composition containing 0.4-20 mg/ml oleanolic acid, said oil phase being one or more of the following: soybean oil, isopropyl myristate, isopropyl palmitate, ethyl laurate, ethyl myristate, glyceryl oleate, glyceryl linoleate, corn oil, sesame oil, olive oil and caprylic/capric glyceride, wherein the content of the oil phase is 30-56%; oleanolic acid is dissolved by the cosurfactant/solvent and is selected from glycerol, polyethylene glycol, propylene glycol, ethylene glycol monoethyl ether, absolute ethanol, dimethylacetamide and N-methylpyrrolidone; in example 22, in which the content of oleanolic acid is up to 20mg/mL, the total surfactant content used is up to 66%, the content of N-methylpyrrolidone (NMP) which is a main solvent is 25%, but N-methylpyrrolidone (NMP) has the harm of irritation, reproductive toxicity and the like, and dimethylacetamide which is a relatively efficient solvent has obvious irritation and toxicity to human bodies and skin, has teratogenicity and is not suitable for long-term use in high concentration.

Chinese patent application No. CN03125480.2 discloses an emulsion for injection of oleanolic acid and a preparation method thereof, comprising: 0.08-2 g/L oleanolic acid, 5-30 g/L injection phospholipid, 50-200 g/L injection soybean oil, 22-25 g/L glycerol and the balance of injection water. The preparation method comprises mixing oleanolic acid and soybean oil to obtain oil phase; mixing and stirring phospholipid, glycerol and water for injection to obtain water phase; adding the oil phase into the water phase under stirring, adjusting the pH to 6.5-7.0, and adding water for injection to obtain crude milk; homogenizing the crude milk with a high pressure homogenizer to obtain emulsion. The emulsion is prepared by adopting the traditional method of mixing and homogenizing oil phase and water phase, and the particle size of the prepared emulsion is larger, about 1 mu m; and the oleanolic acid content is low, the dissolving process is time-consuming, and the process is complex.

Chinese patent CN2011800558173a relates to a soluble cosmetic composition, which uses nanoliposome to embed oleanolic acid to enhance percutaneous absorption of oleanolic acid to promote synthesis of collagen, thereby improving anti-wrinkle effect; the preparation method comprises mixing hydrogenated lecithin, PEG-60 hydrogenated castor oil and other surfactants with polyalcohol, heating to 80deg.C for dissolving, and adding 0.05% oleanolic acid and other additives for mixing; cooling the obtained mixture to 65 ℃, and then adding ethanol into the mixture; uniformly mixing the components of the aqueous portion and heating to 65 ℃ and mixing with the mixture obtained before; the preparation method has the main defects of complex preparation process, multiple heating and cooling requirements, high cost and poor liposome stability; in addition, regardless of how the proportions of surfactant, polyol and water are adjusted, the solvent is limited in its dissolving power, and generally contains only 0.05% oleanolic acid, at a relatively low concentration.

In order to solve the problem of insufficient application of oleanolic acid, the invention uses a pharmaceutical carrier nanoemulsion (nanoemulsion). Nanoemulsions are defined as emulsion systems consisting of an oil phase with a surfactant and water, having an isotropic, transparent (or translucent) appearance. Droplet size is defined as less than 200nm. However, if the emulsion has a low surfactant content and is kinetically stable, dimensions less than 500nm may be accepted as nanoemulsions. These nanocarriers have transparency, low viscosity, thermodynamic stability, ease of preparation, and high surface area making them an effective transdermal and topical drug delivery system; moreover, they have several substantial advantages over other nanocarriers (e.g., microemulsions, liposomes), including low use of surfactants, low skin irritation, high penetration capacity, and high drug loading, and can be used for topical treatment. The nanoemulsion can be used as an initial material for preparing nanocapsules and nanospheres, and can also be prepared into different local formulations, such as foams, gels, creams, liquids and sprays. Although the microemulsion and nanoemulsion are similar in size, their methods of preparation are different. Both of them require energy input, but nanoemulsions mostly use mechanical shear, whereas microemulsions use spontaneous emulsification methods. Microemulsions also require high surfactant concentrations compared to nanoemulsions.

The nanoemulsion gel is formed by adding water-soluble polymer materials such as carbomer, poloxamer, gelatin, cellulose and the like into nanoemulsion to form transparent and stable reticular colloid. The nanoemulsion gel can increase the viscosity of nanoemulsion, improve the adhesiveness and spreadability of nanoemulsion and skin, and the diffusion of the medicine in the gel is blocked due to the increase of the viscosity of the system, the effective diffusion coefficient is reduced, and the volatilization of components is reduced, so that the medicine can maintain longer action time. Is favorable for the full absorption of the medicine, thereby improving the bioavailability of the medicine, being convenient and sanitary to use, being not easy to pollute clothes and having better compliance of patients.

Disclosure of Invention

The invention aims to provide a high-concentration oleanolic acid nanoemulsion composition and a preparation method thereof aiming at the defects existing in the prior art.

Another object of the present invention is to provide the use of the oleanolic acid nanoemulsion composition.

The oleanolic acid nanoemulsion composition comprises the following components in percentage by weight:

0.01 to 8 percent of oleanolic acid, 1 to 40 percent of surfactant, 1 to 40 percent of cosurfactant, 0.2 to 20 percent of terpene-containing essential oil, 0 to 2 percent of baicalin, 0 to 5 percent of oleuropein, a proper amount of additive and the balance of water.

Preferably, the oleanolic acid nanoemulsion composition comprises the following components in percentage by weight:

0.5 to 2 percent of oleanolic acid, 10 to 30 percent of surfactant, 10 to 25 percent of cosurfactant, 1 to 10 percent of terpene-containing essential oil, 0.1 to 1 percent of baicalin, 0.5 to 2 percent of oleuropein, a proper amount of additive and the balance of water.

The oleanolic acid can be pure product or plant extract containing oleanolic acid; the plant extract contains at least 2% oleanolic acid, and can be selected from one of fructus Ligustri Lucidi extract, swertia Japonica Makino extract, herba Swertiae Mileensis extract, fructus Canarii albi extract, etc.

The surfactant is preferably a nonionic surfactant; the surfactant may have an HLB value of from 8 to 18, preferably an HLB value of from 10 to 15; the surfactant may be at least one selected from PEG-6 caprylic/capric glyceride, PEG-8 caprylic/capric glyceride, tween 80, tween 20, polyoxyethylene 40 hydrogenated castor oil, polyoxyethylene castor oil, glyceryl monostearate, span 80, glyceryl monostearate, hydrogenated lecithin, steareth, polyethylene glycol stearate, and polyglyceryl fatty acid ester.

The cosurfactant can be at least one selected from diethylene glycol monoethyl ether, dipropylene glycol, 1, 3-butanediol, ethanol, 1, 2-pentanediol, 1, 2-hexanediol, propylene glycol, glycerol, polyethylene glycol and the like; diethylene glycol monoethyl ether is preferred.

Wherein the terpene containing essential oil contains a terpene structurally belonging to the group of mono-, sesqui-, di-or triterpenes, at least 1% of the terpene, more preferably at least 10% of the terpene, by weight of the terpene containing essential oil. Terpene containing essential oils include, but are not limited to, cedar essential oils, eucalyptus essential oils, peppermint essential oils, tea tree essential oils, lavender essential oils, sage essential oils, clove bud essential oils, cedar essential oils, geranium essential oils, lemon grass essential oils, sage essential oils, turmeric essential oils, wintergreen essential oils, rosemary essential oils, fennel essential oils, chamomile essential oils, guava essential oils, fennel essential oils, basil essential oils, camphor essential oils, kanan, citronella essential oils, cinnamon essential oils, ginger essential oils, perilla essential oils, rose essential oils, patchouli essential oils, sandalwood essential oils, bay essential oils, carrot seed essential oils, bergamot essential oils, grapefruit essential oils, lemon essential oils, citrus essential oils, orange essential oils, oregano essential oils, sweet orange essential oils, thyme essential oils, and other suitable essential oils.

The terpene-containing essential oil is preferably at least one of eucalyptus oil, tea tree essential oil, cedar essential oil, peppermint essential oil, lavender essential oil, rosemary essential oil, etc.; the terpene-containing essential oil comprises, by weight, 0.2% -10% of eucalyptus oil, 0.2% -10% of tea tree essential oil, 0.3% -5% of cedar essential oil, 0.8% -3.2% of peppermint essential oil, 0.6% -2.4% of lavender essential oil and 0.5% -3.2% of rosemary essential oil, preferably: 0.3 to 12 percent of eucalyptus oil, 0.3 to 10 percent of tea tree essential oil, 0.3 to 5 percent of cedar essential oil, 0.8 to 3.2 percent of peppermint essential oil, 0.6 to 2.4 percent of lavender essential oil and 0.5 to 3.2 percent of rosemary essential oil; more preferably: 0.5 to 2 percent of eucalyptus oil, 0.5 to 2 percent of tea tree essential oil, 0.3 to 1 percent of cedar essential oil, 0.8 to 2.0 percent of peppermint essential oil, 0.5 to 1.2 percent of lavender essential oil and 0.5 to 1.6 percent of rosemary essential oil.

The additives may include thickeners, humectants, pH adjusters, preservatives, antioxidants, conditioning agents, chelating agents, ultraviolet absorbers, nutrients, or other suitable additives. Such other suitable additives include, but are not limited to, at least one of an emollient, an anti-androgenic agent, a hair tonic, and the like.

The thickener is selected from carbomer, hydroxyethyl cellulose, sodium carboxymethylcellulose, sodium alginate, polycarbophil, poloxamer, xanthan gum, aloe vera gel, guar gum, polyquaternary ammonium salt Q-10, acrylic acid, and acrylic acid ester; carbomers are preferred; the usage amount of the thickener is preferably 0.1% -5% of the total weight of the nanoemulsion;

the humectant is at least one selected from astragalus polysaccharide, sodium hyaluronate, dipotassium glycyrrhizinate, glycerol, polyethylene glycol, sorbitol, maltitol, aloe vera gel, sodium lactate, squalane, etc.; preferably astragalus polysaccharides and sodium hyaluronate; the usage amount of the humectant is preferably 0.5-5% of the total weight of the nanoemulsion.

The pH adjustor can be selected from the group consisting of citric acid, malic acid, lactic acid, phosphoric acid, triethanolamine, and combinations thereof, preferably triethanolamine; the pH regulator is preferably used in an amount of 0.1-1% of the total weight of the nanoemulsion.

The preservative can be selected from p-hydroxyacetophenone, phenoxyethanol, benzoic acid, octanoyl hydroxamic acid, octanoyl glycol, methyl benzoate, ethyl benzoate, propyl benzoate, butyl benzoate, dimethylaminoethanol, benzalkonium chloride, benzalkonium bromide and the like, and the use amount of the preservative is preferably 0.1-0.6% of the total weight of the nanoemulsion.

The antioxidant is one or more selected from sodium bisulphite, vitamin E, vitamin C, lauryl gallate, propyl gallate, phenethyl caffeate, arbutin, ferulic acid, magnesium ascorbyl phosphate, sodium ascorbyl phosphate and ascorbyl ethyl ether.

The conditioner is one or more of allantoin, dipotassium glycyrrhizinate, tocopheryl acetate, baicalein, olive leaf extract, baical skullcap root extract, lavender extract, chamomile extract, witch hazel extract, purslane extract, gentian root extract and rosemary extract.

The chelating agent is selected from EDTA, EDTA-2Na or EDTA-4Na.

The ultraviolet absorbent is at least one selected from baicalin derivatives, quercetin derivatives, p-aminobenzoic acid derivatives, salicylic acid derivatives, benzophenone derivatives, diphenylacrylic acid esters, cinnamic acid derivatives, etc.

The nutritional agent is at least one selected from lysine, cystine, arginine, collagen, biotin, chondroitin sulfate, keratan sulfate, heparin sulfate, polypeptide, vitamins A, B1, B2, B6, E, C, vitamin C derivatives and the like.

The emollient is selected from cetostearyl alcohol, lanolin, isopropyl isostearate, vaseline, liquid paraffin, polydimethylsiloxane, dimethiconol, dimethicone, shea butter, cetyl isooctanoate, natural squalane, octyl methoxycinnamate.

The antiandrogen agent can be at least one selected from finasteride, dutasteride, irinotecan, bicalutamide, carthamin, equol, soybean isoflavone, ligustrin, hinokitiol, saw palmetto extract, kuh-seng extract, red sage extract, lavender extract, genistein, turmeric extract, phyllanthus emblica extract, nettle extract, speranskia herb extract, ganoderma lucidum extract, dittany bark extract, azelaic acid, quercetin, myricetin, magnolol, luteolin, kaempferol, rutin, tea polyphenol, curcumin, emodin, baicalein, tea tree extract, kudzuvine root extract, genistein, baicalin, cimetidine, cyproterone, hexane estrol, 11-alpha-hydroxy progesterone, norgestrel, levonorethindrone, norethindrone, and the like.

The pilatory is selected from one or more of Polygoni Multiflori radix extract, fructus Ligustri Lucidi extract, carthami flos extract, folium Platycladi extract, herba Rosmarini officinalis extract, ginseng radix extract, notoginseng radix extract, green tea extract, herba Centellae extract, herba Swertiae Mileensis extract, semen Pisi Sativi extract, ginsenoside, radix Angelicae sinensis extract, folium Oleae Europaeae extract, radix astragali extract, rhizoma Ligustici Chuanxiong extract, rhizoma Zingiberis recens extract, semen Sesami Niger extract, semen Cuscutae extract, ecliptae herba extract, folium Mori extract, epigallocatechin gallate (EGCG), cypress wood alcohol, stilbene glucoside, adenosine, caffeine, and minoxidil.

The water may be selected from distilled water, purified water and/or deionized water, preferably distilled water.

The additive of the oleanolic acid nanoemulsion composition of the present invention is usually such that a fat-soluble component is dissolved in the oil phase of the emulsifying system and a hydrophilic component is dissolved in the water phase component of the emulsifying system.

When the dosage form of the present invention is an ointment, paste, cream or gel, as a carrier component, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silica, talc, zinc oxide or mixtures thereof can be used.

When the dosage form of the present invention is a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or mixtures thereof may be used as carrier components, especially when the dosage form is a spray, propellants such as chlorofluorocarbons, propane/butane or dimethyl ether and the like may be further included.

When the dosage form of the present invention is an emulsion, a liquid diluent such as water, ethanol or propylene glycol may be used as the carrier component.

The preparation method of the oleanolic acid nanoemulsion composition comprises the following steps:

1) Adding a formula amount of cosurfactant and terpene-containing essential oil into a container A, and uniformly mixing and stirring; adding surfactant with formula amount, and mixing; then adding oleanolic acid with formula amount, and further adding other fat-soluble components (such as baicalin) and stirring until completely dissolving;

2) Adding water with a formula amount into a container B, and further adding a proper amount of additives to obtain a water phase;

3) Slowly adding the mixture of the container A into the water phase of the container B while stirring to obtain primary emulsion;

4) And 3) performing high-speed shearing emulsification on the primary emulsion prepared in the step 3), and performing secondary emulsification by using a high-pressure homogenizing and emulsifying device, so that the oleanolic acid nanoemulsion composition is obtained after full emulsification.

In the step 1), the mass ratio of the cosurfactant to the terpene-containing essential oil is 4:1-1:1; the dissolution can be stirred at the normal temperature of 25-40 ℃ until the dissolution is completed, or the mixture is heated and stirred at the water bath condition of 40-80 ℃ until the dissolution is completed;

in the step 4), the high-pressure homogenizing and emulsifying device is used for secondary emulsification, the homogenizing speed is 4000-10000 rpm, the homogenizing time is 3-10 min, the high-pressure homogenizing pressure is 800-1000 bar, and the homogenizing is carried out for 3-5 times; the average particle size of the liquid drops in the prepared oleanolic acid nanoemulsion composition can be 10-200 nm, and is preferably less than 100nm; the secondary emulsification can also adopt micro-jet emulsification or ultrasonic emulsification.

The composition of the present invention is used as a nanoemulsion, and the average particle diameter of the droplets is preferably 10 nm to 200 nm. The emulsification process can be selected from the following steps: self-emulsifying, ultrasonic emulsifying, or microfluidizing; if self-emulsifying is used, stirring and uniformly mixing the primary emulsion to obtain nano emulsion; if ultrasonic emulsification is used, the primary emulsion is subjected to ultrasonic treatment at normal temperature, the power is 0.1-75 KW, the frequency is 20 KHz-500 MHz, the time interval is 3s when each ultrasonic wave is 3-5 s, and the whole ultrasonic treatment time is 15-300 s; if the microjet is emulsified, the primary emulsion is added into a microjet high-pressure homogenizer, and is treated for three times by increasing the pressure, the pressure is 1000bar, 1200bar and 1400bar in sequence, and then the obtained solution is filtered by an ultra-micro filter membrane with the aperture of 100 nm.

The oleanolic acid nanoemulsion composition can be applied to preparation of cosmetics. The cosmetics include, but are not limited to, hair growth lotion, hair care essence, skin care gel, anti-aging formulation, shampoo, skin foundation, essence lotion, make-up lotion, toner, acne lotion, emulsion, microemulsion, shampoo, gel, perfume, hair wax, hair conditioner, hair mask, mascara, hair oil, massage oil, hair conditioner, hair blacker, hair dye, hair gel, hair mousse, and the like.

The oleanolic acid nanoemulsion composition is used in a medicament or a cosmetic for improving or preventing alopecia, hair rarefaction, white hair, acne, seborrheic dermatitis, folliculitis, skin aging or other unhealthy skin conditions. The alopecia includes androgenetic alopecia, telogen effluvium, alopecia areata or other types of alopecia.

The oleanolic acid nanoemulsion composition can be used for skin, hair, facial skin care and the like.

The oleanolic acid nanoemulsion composition can be added with an excipient to be used as a dosage form for skin or hair care, and the oleanolic acid nanoemulsion composition is used alone as a product or is mixed with other cosmetic or medicine raw materials to prepare nanoemulsion gel, ointment, cream, emulsion, spray, lotion, liniment, patch or coating dosage form.

In the embodiment of the invention, oleanolic acid and terpene-containing essential oil are taken as active ingredients, and other functional ingredients such as baicalin can be added. Baicalin has effects of resisting skin allergy, relieving inflammation, inhibiting bacteria, preventing corrosion, and resisting oxidation, and can be used for preventing and treating skin allergy. Baicalin can absorb ultraviolet rays, scavenge oxygen free radicals, inhibit melanin generation, antagonize Androgen Receptor (AR) and reduce AR gene expression, and can reduce alopecia and acne. Baicalin is extremely insoluble in various inorganic and organic solvents, and is poor in water solubility and fat solubility, so that the nanoemulsion system can achieve better dissolution and improve bioavailability and biological efficacy.

Examples of nanoemulsions according to the invention as cosmetic and/or pharmaceutical compositions also include oleuropein (oleuropein). Oleuropein is a glycosylated terpenoid, is a main effective component of olive leaf extract, has powerful antibacterial and antiviral properties, and has extremely strong antioxidant capacity. It has been demonstrated that topical application of oleuropein promotes collagen growth and increases VEGF, IGF-1, KGF, HGF expression, and these growth factors are closely related to skin healing and hair growth, thus also promoting hair growth, accelerating wound healing, soothing and calming the skin.

Embodiments of the present invention as cosmetic and/or pharmaceutical compositions may also comprise astragalus polysaccharides. The astragalus polysaccharide has the functions of antioxidation and aging resistance, and has good hygroscopicity, thereby playing a role in moisturizing skin; polysaccharides also have a thickening effect, so that astragalus polysaccharides can be used as a thickening agent; in addition, the astragalus polysaccharide has the effect of promoting hair regeneration and has good synergistic effect with oleanolic acid.

Compared with the prior art, the invention has the outstanding advantages and technical effects that:

1. the particle size of the liquid drops in the oleanolic acid nanoemulsion is nanoscale, the particle size is less than 200nm, the liquid drops are spherical and distributed uniformly, and the defects of larger particle size and poor thermodynamic stability of the traditional emulsion are overcome: the dispersibility of the oleanolic acid in water is obviously improved, the solubility is improved by more than 10000 times, and the bioavailability of the oleanolic acid is improved. The oleanolic acid nanoemulsion composition can greatly improve the solubility and the dissolution rate of oleanolic acid, can also improve the drug loading rate after being prepared into a dosage form, improves the treatment or improvement effect, and is beneficial to the application of oleanolic acid in cosmetics and external pharmaceutical preparations.

2. The positive characteristics (namely physiological activity) of the oleanolic acid are combined with terpene-containing essential oil with good transdermal absorption capacity and nutrition or healing effect, and the combination of the oleanolic acid and the terpene-containing essential oil plays an excellent synergistic effect; the two are combined by intermolecular force, the terpene-containing essential oil enhances the dissolution and percutaneous absorption of oleanolic acid, the oleanolic acid reduces volatilization of the essential oil, and sensitization of the essential oil to individual skin sensitizers is reduced or prevented.

3. The oleanolic acid nanoemulsion is an isotropic, good-fluidity, clear, transparent or semitransparent dispersion system with slightly opalescence formed by oleanolic acid under the combined action of the essential oil, the surfactant and the cosurfactant, can be kept clear after being diluted by water, has good organoleptic properties, can reduce viscosity and greasy feel by taking the essential oil as an oil phase, has natural plant fragrance, and has excellent practical use comfort.

4. Improves the anti-corrosion capability of the product, and reduces or even does not add anti-corrosion additives. The essential oil used by the nanoemulsion has an antibacterial and antiseptic effect, and the oleanolic acid also has a strong antibacterial effect, so that the product deterioration can be prevented without adding a chemical preservative, the skin irritation is reduced, and the safety is improved.

5. Oleanolic acid is required to penetrate the skin to exert its effect, and must be able to penetrate the stratum corneum barrier to reach the corresponding sites of action and maintain a certain effect time at these sites. The nanoemulsion has small particle size, and is beneficial to transdermal absorption and efficacy of the efficacy components; one or more natural plant essential oils with penetration promoting effect are used, chemical penetration promoting agents are not needed to be added, the percutaneous absorption capacity of oleanolic acid and other active ingredients in the composition is improved, and the safety is better; the nanoemulsion contains water, so that the skin has better hydration property and is more favorable for absorption.

6. The preparation process of the product is simple and is beneficial to environmental protection. The traditional emulsion cosmetics or external medicinal preparations have complex oil phase and water phase components, more additives, complicated process flow and longer production period because of the need of using various raw materials, or use toxic and harmful solvents for improving the dissolution amount of oleanolic acid; the preparation process of the invention can be carried out under the room temperature condition, has stable compatibility with active ingredients, can contain essential oil and other additives without denaturing the essential oil and other additives, has simple preparation ingredients, low cost and simple preparation process, is easy for industrial production and reduces environmental pollution.

Drawings

FIG. 1 is a product appearance diagram of oleanolic acid nanoemulsion compositions prepared in examples 1-6.

Fig. 2 is a graph showing the comparison of the groups after experimental example 1 of the effect of oleanolic acid nanoemulsion on androgen alopecia model mice.

Wherein A is a blank group; b is a model group; c is minoxidil group; d is 1% oleanolic acid group (group 4); e is oleanolic acid + oleuropein + baicalin group (group 5).

Detailed Description

The invention will be further illustrated by the following examples in conjunction with the accompanying drawings. The exemplary compositions are merely illustrative and do not limit the scope of the invention. The proportions of the examples and other parts of the specification are by weight unless otherwise indicated. The following formulations are directed to nanoemulsion, gel embodiments; however, similar techniques may also be used in other types of cosmetic or laundry products, including but not limited to shampoos, creams, sprays, conditioners, and styling products.

The invention relates to a nanoemulsion carrier of oleanolic acid. The terpenoid refers to derivatives with a general formula of (C5H 8) n and oxygen content and different saturation degrees, the derivatives are widely existing in the natural world, monoterpenes and sesquiterpenes are widely existing in plant essential oil, and oleanolic acid is a pentacyclic triterpene compound and belongs to terpenes. The solubility of the essential oil in alignment with oleanolic acid is of interest to the inventors based on the principle of similar miscibility; according to the assay of the present invention, many terpene containing plant essential oils have surprising solubilization of oleanolic acid, such as eucalyptus oil, tea tree essential oil, cedar essential oil, lavender essential oil, rosemary essential oil, peppermint essential oil, lemon essential oil, carrot seed oil, and the like; in addition, some essential oils contain a large amount of liquid terpene monomers, such as Eucalyptol (Eucalyptol) of eucalyptus oil and rosemary essential oil, terpin-4-ol (terpin-4-ol) of tea tree oil, linalool (linalol) of lavender essential oil, geraniol (Geraniol) of rose essential oil and the like, which have excellent oleanolic acid solubility.

The applicant has found that nanoemulsions containing terpene containing essential oils as the oil phase are suitable matrices for oleanolic acid. Thus, the positive properties of oleanolic acid are combined with nanoemulsions comprising terpene containing essential oils. In particular, nanoemulsions can be produced with high oleanolic acid loading, which combine the positive properties of oleanolic acid (i.e. physiological activity) with terpene containing essential oil having good transdermal absorption capacity and nutritional or healing effects, if any. This makes the nanoemulsion formulation particularly suitable for cosmetic or dermatological preparations of triterpenes.

The preparation method is characterized in that a mixture of a cosurfactant and terpene-containing essential oil in a mass ratio of 4:1-1:1 is used for solubilizing oleanolic acid: not only can prevent coagulation from occurring in the middle of dissolution, but also can greatly increase the concentration of oleanolic acid in the preparation.

In embodiments of the present invention, novel utilities and/or advantages may be provided when the essential oils are both carriers and solvents and active ingredients; the invention seals the essential oil in the nano system as the oil phase, can improve the solubility of oleanolic acid, and plays a synergistic effect with the active ingredients in application.

Essential oils in embodiments of the present invention have a number of valuable properties that can be used as combination therapies; plant essential oils are useful in dermatological patients, and they generally have antibacterial, antifungal, anti-inflammatory, astringent and regenerating properties; can be used for aromatherapy for treating dermatoses such as dandruff, psoriasis, eczema, acne, and seborrheic dermatitis; can have very beneficial synergistic effects as active ingredients and/or enhancers in the intended application, in particular as androgen antagonists and hair growth promoters; essential oils such as eucalyptus oil, peppermint oil, tea tree oil and the like have been widely studied and reported as penetration enhancers to increase transdermal drug delivery; in addition, the encapsulation of the nano system can provide the controlled release of bioactive components such as oleanolic acid and the like, and provide better stability of essential oil and other volatile compounds, and improve the bioavailability and biological efficacy of the preparation.

And from a toxicological point of view, essential oils are considered safer and less irritating than other chemical permeation enhancers; essential oils or fixed oils extracted from aromatic plants are widely used in foods, cosmetics and pharmaceuticals, many of which are listed in the us FDA (us food and drug administration) list of Generally Recognized As Safe (GRAS), i.e. "acknowledged safety compounds", essential oils, oleoresins (without solvents) and natural extracts (including distillates) are generally considered safe for their intended use (foods, cosmetics), such as menthol, peppermint, rosemary, sage, orange flowers, lemon GRASs, thyme, orange leaves, citronella, ylang, etc., and are also suitable for long-term use without risk.

Eucalyptus oil, also called as UGANLI essential oil, has antibacterial, antioxidant, antiinflammatory, permeation promoting, insecticidal, mosquito repellent, antipruritic, and antiseptic effects, is a good raw material for medicine and perfume, and is widely used in food processing and cosmetics. Eucalyptus oil is also used to improve blood circulation of the scalp, thereby promoting hair growth.

The tea tree essential oil has powerful cleaning, antibacterial, anti-inflammatory and antiseptic properties, and is suitable for oily and acne skin, and can be used for treating suppurative wound, burn, sunburn, psoriasis, dandruff, etc. In addition, tea tree essential oil has weak estrogen-like activity, and in vitro studies prove that the expression of androgen receptor can be reduced, so that androgenetic alopecia can be reduced.

It is well known that peppermint essential oil can produce a cooling sensation and promote blood circulation, helps reduce skin inflammation and itching, and has been demonstrated to promote hair growth.

Cedrol, the main ingredient of cedar essential oil, is also the main active ingredient of cacumen Platycladi, and has vasodilating effect on blood vessel, and can stimulate hair follicle to generate new hair growth, and inhibit natural shedding process. In addition, the cedar essential oil has strong antifungal and antibacterial properties, and the addition of the cedar essential oil in the treatment of acne helps to relieve and reduce intractable acne outbreaks.

Lavender essential oils have been shown to have weak estrogenic activity and very strong anti-inflammatory properties, and can play a key role in helping users relieve itching, swelling of the skin, and increase the rate of wound healing, thereby providing a good environment for skin repair and hair growth.

The rosemary essential oil has strong astringing effect, improves greasy desquamation skin, promotes blood circulation, and can stimulate hair regeneration.

In some embodiments, the composition may contain other additives such as thickeners, humectants, pH adjusting agents, stabilizers, preservatives, antioxidants, conditioning agents, emollients, colorants, skin protectants, skin and hair nutrients such as collagen, elastin, keratin, and other suitable additives.

In some embodiments, the composition comprises a humectant selected from one or more of astragalus polysaccharides, sodium hyaluronate, dipotassium glycyrrhizinate, glycerin, aloe vera gel, more preferably astragalus polysaccharides and sodium hyaluronate;

in some embodiments, the composition may further comprise a thickening agent, such as a naturally derived thickening agent, e.g., carbomer, hydroxyethyl cellulose, sodium carboxymethyl cellulose, sodium alginate, poloxamer 407, xanthan gum, guar gum, more preferably carbomer.

In some embodiments, the composition may further comprise a pH adjuster, which may be selected from the group consisting of citric acid, malic acid, lactic acid, phosphoric acid, triethanolamine, and combinations thereof, preferably triethanolamine.

In some embodiments, the composition may not need to be added with a preservative, for example, the essential oil and 1, 3-butanediol, ethanol, 1, 2-pentanediol, 1, 2-hexanediol all have antiseptic and bacteriostatic effects; in some embodiments, small amounts of preservatives including p-hydroxyacetophenone, phenoxyethanol may be optionally used.

The compositions of the present invention are hydrophilic in nature. The water may be distilled, purified and/or deionized, preferably distilled water. In some embodiments, the composition comprises about 1% to 80% water by weight.

The average particle size of the liquid drops in the oleanolic acid nanoemulsion composition is 10-200 nm, and the particle size of the nanoemulsion is preferably smaller than 100nm. The application process can be one or more of high-pressure homogenization in high-energy emulsification, micro-jet or ultrasonic, and high-pressure homogenization is preferred.

The obtained nanoemulsion can be used alone, or can be further added with other excipients to prepare nanoemulsion gel, cream, emulsion, spray, transdermal patch, ointment and the like.

According to the preparation method provided by the invention, terpene-containing essential oils such as eucalyptus oil, tea tree oil and cedar oil have higher solubility in oleanolic acid, however, when the dissolution amount of oleanolic acid in the essential oils exceeds 40mg/mL, white colloidal coagulation is easy to form, and further dissolution is seriously hindered.

The preparation method is characterized in that a mixture of a cosurfactant and terpene-containing essential oil in a mass ratio of 4:1-1:1 is used for solubilizing oleanolic acid: not only can prevent coagulation in the middle of dissolution, but also can greatly increase the concentration of oleanolic acid in the preparation;

in embodiments of the present invention, novel utilities and/or advantages may be provided when the essential oils are both carriers and solvents and active ingredients; the invention seals the essential oil in the nano system as a lipophilic phase, can improve the solubility of oleanolic acid, and plays a synergistic effect with active ingredients in application.

The oleanolic acid nanoemulsion composition of the present invention may further comprise other hydrophilic or fat-soluble active ingredients, thereby enabling cosmetics or external preparations to have a stronger activity. In general, the fat-soluble active ingredient is dissolved in the oil phase of the emulsifying system and the hydrophilic ingredient is dissolved in the water phase of the emulsifying system. The addition of active ingredients such as baicalin with antiandrogenic action, other active ingredients such as oleuropein, astragalus polysaccharides should be noted. The formula of the nanoemulsion composition is embodied in dosage forms such as nanoemulsion, essence, gel or shampoo, and therefore, the nanoemulsion also comprises auxiliary components such as thickening agents, moisturizing agents, preservatives and the like.

In the embodiment of the invention, the oleanolic acid and the terpene-containing essential oil are taken as active ingredients, and other active ingredients and/or reinforcing agents such as baicalin can be added. Other active ingredients and/or enhancers such as baicalin may also be added to embodiments of the present invention. Baicalin is known to have the effects of resisting skin allergy, resisting inflammation, inhibiting bacteria, preserving and resisting oxidation, and can be used for preventing and treating skin allergy. Baicalin can absorb ultraviolet rays, scavenge oxygen free radicals, inhibit melanin generation, antagonize Androgen Receptor (AR) and reduce AR gene expression, and can reduce alopecia and acne. Baicalin is extremely insoluble in various inorganic and organic solvents, and is poor in water solubility and fat solubility, so that the nanoemulsion system can achieve better dissolution and improve bioavailability and biological efficacy.

Embodiments of the present invention as cosmetic and/or pharmaceutical compositions may also comprise astragalus polysaccharides. The astragalus polysaccharide has the functions of antioxidation and aging resistance, and has good hygroscopicity, thereby playing a role in moisturizing skin; polysaccharides also have a thickening effect, so that astragalus polysaccharides can be used as a thickening agent; in addition, the astragalus polysaccharide has the effect of promoting hair regeneration and has good synergistic effect with oleanolic acid. Other plant extracts with hair growth promoting and antiandrogenic effects, such as ginsenoside, ginseng radix extract, polygoni Multiflori radix extract, herba Rosmarini officinalis extract, carthami flos extract, platycladus orientalis extract, scutellariae radix extract, etc. can be used in the preparation of the present invention.

Because the quality standards of the same functional components and raw materials are not identical at present, and particularly the content and purity of the essential oil components are different, the solubility of the active components can be influenced, and therefore, great attention is paid to the quality and source of the raw materials when the product is prepared. In the embodiment of the invention, the content of eucalyptol in eucalyptus oil is more than or equal to 80%, the content of terpin-4-ol in tea tree oil is more than or equal to 42%, the content of linalool in lavender essential oil is more than or equal to 31%, the content of eucalyptol in rosemary essential oil is more than or equal to 26%, the content of cedar essential oil is more than or equal to 11%, the content of peppermint essential oil is peppermint essential oil, and the content of menthol is more than or equal to 36%; the purity of diethylene glycol monoethyl ether is more than or equal to 99.5%, and is cosmetic grade or medical grade, preferably Transcutol CG or Transcutol HP.

Specific examples are given below.

Example 1:

the 1% oleanolic acid nanoemulsion comprises the following raw material components in percentage by weight as shown in table 1:

TABLE 1

Component (A)	Weight percent (%)
		Oleanolic acid	1
Eucalyptus oil	1
		Tea tree oil	2
Rosemary essential oil	1.2
		Peppermint essential oil	2
Lavender essential oil	0.8
		Diethylene glycol monoethyl ether (Transcutol)	16
Dipropylene glycol	5
		Tween 20	25.00
Polyoxyethylene 35 castor oil	8
		Distilled water	37.8

The preparation method comprises the following steps:

(1) and weighing eucalyptus oil, tea tree oil, rosemary essential oil, peppermint essential oil and lavender essential oil according to the formula amount, and uniformly mixing the eucalyptus oil, the tea tree oil, the rosemary essential oil, the peppermint essential oil and the lavender essential oil with diethylene glycol monoethyl ether and dipropylene glycol in a container A.

(2) Weighing Tween 20 and polyoxyethylene 35 castor oil, adding into the mixture of container A, mixing, adding oleanolic acid, and stirring to dissolve completely.

(3) Distilled water was added to vessel B followed by mixing well.

(4) Adding the mixture of the container A into the container B under stirring to obtain primary emulsion, and then emulsifying for the second time by using a high-pressure homogenizing emulsifying device, wherein the homogenizing speed is 5000rpm, the homogenizing time is 5min, the high-pressure homogenizing pressure is 800bar, and homogenizing is carried out for 3-5 times.

(5) And (3) closing the high-pressure homogenizer, and taking a small amount of finished products for observation, wherein the appearance is transparent or semitransparent, the fluidity and the dispersibility are good, and the oleanolic acid nanoemulsion is obtained when the oleanolic acid nanoemulsion has obvious visible opalescence.

Example 2:

the 1% oleanolic acid nanoemulsion comprises the following raw material components in percentage by weight as shown in table 2:

TABLE 2

Component (A)	Weight percent (%)
		Oleanolic acid	1
Eucalyptus oil	2
		Tea tree oil	2
Lavender essential oil	1.2
		Peppermint essential oil	1.8
Dipropylene glycol	5
		Ethanol	3
1, 2-hexanediol	8
		PEG-6 caprylic/capric glyceride	16
Polyoxyethylene 35 castor oil	12
		Biotin	0.5
Dipotassium glycyrrhizinate	0.5
		Distilled water	47

The preparation method comprises the following steps:

(1) the eucalyptus oil, tea tree oil, peppermint essential oil, lavender essential oil, 1, 2-hexanediol, ethanol, dipropylene glycol, PEG-6 caprylic/capric glyceride and polyoxyethylene 35 castor oil with the formula amounts are weighed and evenly mixed in a container A.

(2) The oleanolic acid with the formula amount is weighed and added into the mixture of the container A, and the mixture is placed in a water bath with the temperature of 80 ℃ for heating and stirring until the oleanolic acid is completely dissolved.

(3) Distilled water is added into the container B, and biotin and dipotassium glycyrrhizinate are added and stirred until dissolved.

(4) Adding the mixture of the container A into the container B under stirring to obtain primary emulsion, and then emulsifying for the second time by using a high-pressure homogenizing emulsifying device, wherein the homogenizing speed is 4000rpm, the homogenizing time is 5min, the high-pressure homogenizing pressure is 800bar, and homogenizing is carried out for 3-5 times.

(5) And (3) closing the high-pressure homogenizer, and taking a small amount of finished products for observation, wherein the appearance is colorless, the fluidity and the dispersibility are good, and the oleanolic acid nanoemulsion is obtained if the appearance is obvious in opalescence.

Example 3:

the 5% oleanolic acid nanoemulsion comprises the following raw material components in percentage by weight as shown in table 3:

TABLE 3 Table 3

Component (A)	Weight percent (%)
		Oleanolic acid	5
Eucalyptus oil	12
		Tea tree oil	8
Diethylene glycol monoethyl ether (Transcutol)	36
		PEG-8 caprylic/capric glyceride	36
Distilled water	3

The preparation method comprises the following steps:

(1) the eucalyptus oil, tea tree oil, diethylene glycol monoethyl ether (Transcutol) and PEG-8 caprylic/capric glyceride with the formula amounts are weighed and added into a container A to be uniformly mixed.

(2) And (3) adding oleanolic acid with the formula amount into the mixture, and heating and stirring in a water bath at 60 ℃ until the oleanolic acid is completely dissolved.

(3) Distilled water was added to vessel B, and then the mixture of vessel a was slowly added to vessel B with stirring to obtain a primary emulsion.

(4) And then using a high-pressure homogenizing and emulsifying device to perform secondary emulsification, wherein the homogenizing speed is 4000rpm, the homogenizing time is 3min, the high-pressure homogenizing pressure is 800bar, and the homogenizing is performed for 3-5 times.

Example 4:

the oleanolic acid nanoemulsion for promoting hair growth comprises the following raw material components in percentage by weight as shown in table 4:

TABLE 4 Table 4

Component (A)	Weight percent (%)
		Oleanolic acid	1
Oleuropein	1.6
		Baicalin	0.5
Tea tree oil	2
		Eucalyptus oil	1
Rosemary essential oil	1.2
		Peppermint essential oil	1.6
Lavender essential oil	0.4
		Cedar essential oil	0.2
1,2 pentanediol	5
		Diethylene glycol monoethyl ether (Transcutol)	23
Astragalus polysaccharides	1
		EDTA-4Na	0.1
Para hydroxy acetophenone	0.3
		Arginine (Arg)	0.6
Polyoxyethylene 30 castor oil	7
		Tween 80	12
Polyoxyethylene 40 hydrogenated castor oil	16
		Distilled water	25.5

The preparation method comprises the following steps:

(1) the eucalyptus oil, tea tree oil, peppermint essential oil, lavender essential oil, rosemary essential oil, cedar essential oil, dipropylene glycol and diethylene glycol monoethyl ether (Transcutol) are respectively weighed according to the formula amount and uniformly mixed in a container A.

(2) Weighing polyoxyethylene 30 castor oil, tween 80 and polyoxyethylene 40 hydrogenated castor oil, mixing, adding into the mixture of container A, adding oleanolic acid and baicalin, heating in 80deg.C water bath, and stirring to dissolve completely.

(3) Adding distilled water into container B, adding oleuropein and/or Astragalus polysaccharides, EDTA-4Na, arginine and p-hydroxyacetophenone, and stirring to dissolve.

(5) And (3) closing the high-pressure homogenizer, and taking a small amount of finished products for observation, wherein the appearance is brown, semitransparent, fluid and dispersive, and the oleanolic acid nanoemulsion is obtained when the oleanolic acid nanoemulsion has obvious visible opalescence.

Example 5:

the oleanolic acid skin care nanoemulsion comprises the following raw material components in percentage by weight as shown in table 5:

TABLE 5

Component (A)	Weight percent (%)
		Oleanolic acid	0.5
Oleuropein	1
		Baicalin	0.2
Tea tree oil	0.7
		Uganli essential oil	0.5
Peppermint essential oil	1.2
		Peppermint essential oil	0.5％
Lemon essential oil	0.2％
		1, 2-hexanediol	3
Dipropylene glycol	3
		Diethylene glycol monoethyl ether (Transcutol)	8
PEG-8 caprylic/capric glyceride	12
		Polyoxyethylene 40 hydrogenated castor oil	15
Glyceryl monostearate	3
		Sodium hyaluronate	1
Vitamin C	0.7
		Aloe vera gel	0.3
Phenoxyethanol	0.3
		EDTA-2Na	0.1
Distilled water	57

The preparation method comprises the following steps:

(1) tea tree oil, uganli essential oil, lemon essential oil, 1, 2-hexanediol, dipropylene glycol and diethylene glycol monoethyl ether (Transcutol) are weighed according to the formula amount respectively and uniformly mixed in a container A.

(2) The polyoxyethylene 30 castor oil, the Tween 80 and the polyoxyethylene 40 hydrogenated castor oil with the formula amounts are weighed and added into the mixture of the container A, and then oleanolic acid and baicalin are added and placed in a water bath with the temperature of 60 ℃ for heating and stirring until the mixture is completely dissolved.

(3) Adding distilled water into container B, adding oleuropein, phenoxyethanol, EDTA-2Na, and vitamin C, stirring to dissolve, adding sodium hyaluronate and aloe gel, standing for 24 hr, and swelling.

(4) Adding the mixture of the container A into the container B under stirring to obtain primary emulsion, and then emulsifying for the second time by using a high-pressure homogenizing emulsifying device, wherein the homogenizing speed is 6000rpm, the homogenizing time is 5min, the high-pressure homogenizing pressure is 800bar, and homogenizing is 3-5 times.

(5) And (3) closing the high-pressure homogenizer, and taking a small amount of finished products for observation, wherein the appearance is light yellow, semitransparent, good in fluidility and dispersibility, and obvious in visible opalescence, namely the oleanolic acid nanoemulsion.

Example 6:

the oleanolic acid acne-removing nanoemulsion gel comprises the following raw material components in percentage by weight as shown in table 6:

TABLE 6

The preparation method comprises the following steps:

(1) the tea tree oil, the peppermint essential oil, the rosemary essential oil, the cedar essential oil, the 1, 3-butanediol and the diethylene glycol monoethyl ether (Transcutol) with the formula amounts are respectively weighed and uniformly mixed in a container A.

(2) Weighing polyoxyethylene 35 castor oil and tween 80, adding into the mixture of container A, adding oleanolic acid and baicalin, heating in 80deg.C water bath, and stirring to dissolve completely.

(3) Distilled water is added into a container B, phenoxyethanol, EDTA-2Na and dipotassium glycyrrhizinate are added, and stirring is carried out until dissolution.

(4) Adding the mixture of the container A into the container B under stirring to obtain primary emulsion, then emulsifying for the second time by using a high-pressure homogenizing emulsifying device, wherein the homogenizing speed is 4000rpm, the homogenizing time is 6min, the high-pressure homogenizing pressure is 800bar, and homogenizing for 3-5 times to obtain the nanoemulsion.

(5) Slowly adding carbomer 940 into the nanoemulsion under stirring, adding triethanolamine, and standing at normal temperature for 24 hr to fully swell carbomer 940 to obtain transparent or semitransparent olea acne-removing nanoemulsion gel.

The product appearance of the oleanolic acid nanoemulsion compositions prepared in examples 1 to 6 is shown in fig. 1.

Test example 1: effect experiment of oleanolic acid nanoemulsion on androgen alopecia model mice

1.1 laboratory animals

40 male C57BL/6 mice are selected, and the weight is 16-18 g, and the age is 3-4 weeks. All rats are placed in an animal house with proper temperature and humidity for feeding, and ventilation is carried out for 1-2 times a day, and each time is 1h. Daily 12 hours of day and night alternation was ensured while keeping quiet and avoiding other disturbances. Ensuring that the mice have enough activity space. The mice are fed with the maintenance feed, purified water is used as drinking water for the rats, and the drinking water and the feed are replaced every day, so that the ventilation, the sanitation and the sanitation of the living environment of the animals are maintained. Illumination is carried out in the daytime (12+/-2) h, the indoor temperature (20+/-2) DEG C and the relative humidity is 40% -60%.

1.2 grouping

40 rats were randomly divided into 5 groups, group 1: blank control 8; group 2: model group 8; group 3: minoxidil group 8; group 4: 8 oleanolic acid groups; group 5: oleanolic acid + oleuropein + baicalin group 8.

1.3 Experimental drugs

The minoxidil group used a commercially available 5% minoxidil solution; the nanoemulsion prepared in example 1 was used for the oleanolic acid group, and the nanoemulsion prepared in example 4 was used for the oleanolic acid, oleuropein and baicalin group.

1.4 moulding

All mice were injected subcutaneously 1 time per day on the back with testosterone propionate at a dose of 5 mg/kg/day for 4 weeks, except for the placebo group, which was injected with an equivalent amount of physiological saline at the same location on the back of the mice.

1.5 administration

After the 4-week molding is successful, normal saline is injected into the blank control group in a subcutaneous mode, testosterone propionate is injected into the other groups continuously for 4 weeks, and the androgen level in the mice is maintained. The model group only continues to inject testosterone propionate 5 mg/kg/day, group 3 uniformly smears 5% minoxidil after each injection of testosterone propionate for 1h, group 4 smears 1% oleanolic acid nanoemulsion prepared in example 1, and group 5 smears oleanolic acid nanoemulsion composition prepared in example 4.

1.6 observations of laboratory animals

1.6.1 general State observation

The unhairing degree of each group of rats is observed every day, and the local skin reactions such as red swelling, edema, liquid seepage and the like do not appear in the medicine smearing area.

1.6.2 preparation of skin tissue sections

After 8 weeks of experiment, the back dehairing area of each group of mice was selected, the parallel spinal column was sampled to about 1x 1m2, and 3 skin tissue sections of substantially the same area were sampled per mouse. Specimens were prepared according to the conventional skin histopathological process, paraffin embedded, sectioned, HE stained, observed under a microscope for morphological changes in hair follicles of mice and counted for total hair follicle number in 100 fields, and the average of three sections was taken for relevant statistical analysis, and the data were expressed as x±sd. When p <0.05, the difference is significant, and has statistical significance.

1.7 experimental results

1.7.1 section follicular observation and follicular counting

Observing the cross section condition of skin hair follicles, and after the external application of the medicine for 4 weeks, the hair of a blank control group is complete and dense, and black and glossy; the area of alopecia of the model group is obviously increased compared with that of the blank group; both minoxidil group and group 3, group 4 mice showed significantly reduced areas of hair loss compared to the model group, see figure 2.

TABLE 7 comparison of follicle count for each group

Group of	Total hair follicle count
		Blank control group (group 1)	38.75±4.12
Model group (group 2)	13.31±3.42*
		Minoxidil group (group 3)	30.38±8.32 ^#
Oleanolic acid set (set 4)	31.63±6.73 ^&
		Oleanolic acid + oleuropein + baicalin group (group 5)	34.12±8.85 ^#$

Note that p compared to the blank control group<0.05； ^# Compared with the molding group, p<0.05；&Compared with minoxidil group, p>0.05； ^$ Compared with minoxidil group, p<0.05。

As can be seen from table 7, oleanolic acid group (group 4) had clear therapeutic effect on androgenic alopecia model mice, and there was no significant statistical difference (P > 0.05) in effect compared to 5% minoxidil group effect; the combined effect of group 4 oleanolic acid + oleuropein + baicalin on promoting hair growth and reducing hair loss in Yu Xiong hormonal hair loss mice was superior to minoxidil group (P <0.05, table 7).

Test example 2 stability investigation of oleanolic acid nanoemulsion formulations

1) Influence of centrifugal acceleration test on nanoemulsion stability

Taking the prepared nano emulsion, centrifuging at 20000r/min, observing that the appearance of the nano emulsion is still clear and transparent after 15min, and avoiding the phenomenon of oil-water delamination.

2) Light stability test

The nanoemulsion of the invention is properly filled into a glass bottle, and is placed at room temperature under the illumination condition of (5000+/-500) lx after being sealed, and is respectively sampled and observed for 1d, 3d and 7 d. The result shows that the nanoemulsion keeps clear and transparent appearance, and has no phenomena of layering, demulsification and the like.

3) Sample retention observation test

The emulsion of the invention is packaged in glass bottles, sealed and then is respectively placed at the temperature of 4 ℃, 25 ℃ and 40 ℃ for sample reserving and inspection for 6 months, and the samples are taken and observed every 14 d. The results show that the emulsion keeps clear and transparent appearance under the four temperature conditions, and no phenomena such as layering and demulsification are seen.

Test example 3: animal skin irritation investigation of oleanolic acid nanoemulsion formulations

New Zealand white rabbits are more responsive to irritants than humans. Skin irritation experiments were performed on examples 4 to 6 by taking 12 healthy New Zealand white rabbits, each half of male and female, and dividing them into 4 groups of 3 each, each half of male and female. The animals were dehaired on both sides of the spinal column 24h before dosing with an electric razor and the dehairing area was 5 x 5cm. The skin irritation test results of the animal are non-irritating, which indicates that oleanolic acid essential oil nanoemulsion and nanoemulsion gel have no obvious irritation to New Zealand white rabbits.

Test example 4: solubilization of oleanolic acid by essential oils

Adding appropriate amount of oleanolic acid into 5mL centrifuge tube, adding 1g of each adjuvant respectively, performing ultrasonic treatment for 30min, balancing at 60deg.C under 100r/min, centrifuging for 10min at 10000r/min, collecting supernatant, diluting filtrate with methanol to appropriate ratio, and performing HPLC to calculate oleanolic acid content in each adjuvant.

The solubility of oleanolic acid in different solvents is shown in Table 8, and from Table 8, the solubility of the oleanolic acid in the essential oil is far higher than that of the common solvents, and the combination of the essential oil and the cosurfactant has better solubilization and dissolution assisting effects on the oleanolic acid.

Table 8: oleanolic acid solubility (mg/ml) of different solvents

Experiments have demonstrated that the topical compositions of the present invention can be used in the preparation of products for the treatment and control of adverse effects of skin conditions including, but not limited to, androgenic alopecia, telogen effluvium, alopecia areata, or other types of alopecia, acne, seborrheic dermatitis, folliculitis, skin aging, and other skin conditions.

The oleanolic acid nanoemulsion composition can be applied to cosmetics and can be used for skin care, hair care and the like.

The oleanolic acid nanoemulsion composition can be used as a dosage form for caring skin or hair by adding an excipient, and can be used independently as a product or mixed with other cosmetics or medicine raw materials to prepare nanoemulsion gel, ointment, cream, emulsion, spray, lotion, liniment, patch or coating.

The oleanolic acid nanoemulsion composition of the present invention can be made into products including, but not limited to, hair growth lotion, hair care essence, skin care gel, shampoo, hair conditioner, mascara, mask, face cream, and lotion.

The above-described embodiments are merely preferred embodiments of the present invention and should not be construed as limiting the scope of the present invention. All equivalent changes and modifications within the scope of the present invention are intended to be covered by the present invention.

Claims

1. The high-concentration oleanolic acid nanoemulsion composition is characterized by comprising the following components in percentage by weight:

0.5 to 2 percent of oleanolic acid, 10 to 30 percent of surfactant, 10 to 25 percent of cosurfactant, 1 to 10 percent of terpene-containing essential oil, 0.1 to 1 percent of baicalin, 0.5 to 2 percent of oleuropein, a proper amount of additive and the balance of water; the terpene-containing essential oil is selected from at least five of eucalyptus oil, tea tree essential oil, cedar essential oil, peppermint essential oil, lavender essential oil and rosemary essential oil;

the surfactant is at least one selected from PEG-6 caprylic acid/capric acid glyceride, PEG-8 caprylic acid/capric acid glyceride, tween 80, tween 20, polyoxyethylene castor oil, span 80, glyceryl monostearate, lecithin, stearyl alcohol polyether and polyethylene glycol stearate;

The cosurfactant is at least one selected from diethylene glycol monoethyl ether, dipropylene glycol, 1, 3-butanediol, ethanol, 1, 2-pentanediol, 1, 2-hexanediol, propylene glycol, glycerol and polyethylene glycol.

2. The high-concentration oleanolic acid nanoemulsion composition of claim 1, wherein the oleanolic acid is pure or a plant extract containing oleanolic acid; the plant extract contains at least 2% oleanolic acid, and is selected from one of fructus Ligustri Lucidi extract, herba Swertiae Mileensis extract, and fructus Oleae Europaeae extract;

the HLB value of the surfactant is 8-18.

3. The high-concentration oleanolic acid nanoemulsion composition of claim 1, characterized in that the terpene-containing essential oil comprises, by weight, 0.2% -10% of eucalyptus oil, 0.2% -10% of tea tree essential oil, 0.3% -5% of cedar essential oil, 0.8% -3.2% of peppermint essential oil, 0.6% -2.4% of lavender essential oil and 0.5% -3.2% of rosemary essential oil.

4. The high concentration oleanolic acid nanoemulsion composition of claim 1, characterized in that the additive comprises at least one of a thickener, a humectant, a pH adjustor, a preservative, an antioxidant, a conditioner, a chelating agent, an ultraviolet absorber, a nutritional agent, an emollient, an antiandrogen agent, a hair tonic;

The thickener is selected from carbomer, hydroxyethyl cellulose, sodium carboxymethylcellulose, sodium alginate, polycarbophil, poloxamer, xanthan gum, aloe gum, guar gum, polyquaternary ammonium salt Q-10, acrylic acid and acrylic ester; the usage amount of the thickener is 0.1% -5% of the total weight of the nanoemulsion;

the humectant is at least one selected from astragalus polysaccharide, sodium hyaluronate, dipotassium glycyrrhizinate, glycerol, polyethylene glycol, sorbitol, maltitol, aloe vera gel, sodium lactate and squalane; the usage amount of the humectant is 0.5% -5% of the total weight of the nanoemulsion;

the pH adjustor is selected from the group consisting of citric acid, malic acid, lactic acid, phosphoric acid, triethanolamine, and combinations thereof; the usage amount of the pH regulator is 0.1% -1% of the total weight of the nanoemulsion;

the preservative is selected from p-hydroxyacetophenone, phenoxyethanol, benzoic acid, octanoyl hydroxamic acid, octanoyl glycol, methyl hydroxybenzoate, ethyl hydroxybenzoate, propyl hydroxybenzoate, butyl hydroxybenzoate, dimethylaminoethanol, benzalkonium chloride and benzalkonium bromide, and the use amount of the preservative is 0.1-0.6% of the total weight of the nanoemulsion;

the antioxidant is one or more selected from sodium bisulphite, vitamin E, vitamin C, lauryl gallate, propyl gallate, phenethyl caffeate, arbutin, ferulic acid, magnesium ascorbyl phosphate, sodium ascorbyl phosphate and ascorbyl ethyl ether;

The conditioner is one or more of allantoin, dipotassium glycyrrhizinate, tocopheryl acetate, olive leaf extract, radix Scutellariae extract, lavender extract, chamomile extract, witch hazel extract, purslane extract, gentian root extract and rosemary extract;

the chelating agent is selected from EDTA, EDTA-2Na or EDTA-4Na;

the ultraviolet absorbent is at least one selected from baicalin derivatives, quercetin derivatives, p-aminobenzoic acid derivatives, salicylic acid derivatives, benzophenone derivatives, diphenyl acrylate and cinnamic acid derivatives;

the nutritional agent is at least one selected from lysine, cystine, arginine, collagen, biotin, chondroitin sulfate, keratan sulfate, heparin sulfate, vitamins A, B1, B2, B6, E, C and vitamin C derivatives;

the emollient is selected from cetostearyl alcohol, lanolin, isopropyl isostearate, vaseline, liquid paraffin, polydimethylsiloxane, dimethiconol, simethicone, shea butter, cetyl isooctanoate, natural squalane, octyl methoxycinnamate;

the antiandrogen agent is at least one selected from finasteride, dutasteride, irinotecan, bicalutamide, equol, saw palmetto extract, kuh-seng extract, red sage extract, lavender extract, turmeric extract, emblic extract, nettle extract, garden balsam extract, ganoderma lucidum extract, dittany bark extract, tea tree extract, kudzu root extract, baical skullcap root extract, cimetidine, cyproterone, diethylstilbestrol, 11-alpha-hydroxyprogestrel, norgestrel, levonorethindrone, norethindrone;

The pilatory is selected from at least one of Polygoni Multiflori radix extract, fructus Ligustri Lucidi extract, carthami flos extract, platycladi extract, herba Rosmarini officinalis extract, ginseng radix extract, notoginseng radix extract, green tea extract, herba Centellae extract, herba Swertiae Mileensis extract, semen Pisi Sativi extract, ginsenoside, radix Angelicae sinensis extract, folium Oleae Europaeae extract, radix astragali extract, rhizoma Ligustici Chuanxiong extract, rhizoma Zingiberis recens extract, semen Sesami Niger extract, semen Cuscutae extract, ecliptae herba extract, and folium Mori extract.

5. The high concentration oleanolic acid nanoemulsion composition of claim 1, wherein the water is selected from distilled water, purified water, or deionized water.

6. The method for preparing the oleanolic acid nanoemulsion composition of claim 1, characterized by comprising the steps of:

1) Adding a formula amount of cosurfactant and terpene-containing essential oil into a container A, and uniformly mixing and stirring; adding surfactant with formula amount, and mixing; then adding oleanolic acid with formula amount, and further adding baicalin with other fat-soluble components, stirring until completely dissolving;

7. The method for preparing the oleanolic acid nanoemulsion composition of claim 6, characterized in that in step 1), the mass ratio of the cosurfactant to the terpene-containing essential oil is 4:1 to 1:1; the dissolution is to stir at normal temperature of 25-40 ℃ until the dissolution is complete, or to stir at 40-80 ℃ in water bath condition until the dissolution is complete;

in the step 4), the high-pressure homogenizing and emulsifying device is used for secondary emulsification, the homogenizing speed is 4000-10000 rpm, the homogenizing time is 3-10 min, the high-pressure homogenizing pressure is 800-1000 bar, and the homogenizing is carried out for 3-5 times; the average particle diameter of the liquid drops in the prepared oleanolic acid nanoemulsion composition is 10-200 nm; the secondary emulsification adopts micro-jet emulsification or ultrasonic emulsification.

8. The use of the oleanolic acid nanoemulsion composition of claim 1 in the preparation of cosmetics; the cosmetic includes, but is not limited to, hair tonic, shampoo, gel, perfume, hair wax, hair conditioner, hair mask, mascara, hair oil, hair conditioner, hair blackening agent, hair dye, hair gel, hair mousse;

The oleanolic acid nanoemulsion composition is used in cosmetics for improving or preventing alopecia, sparse hair and white hair; the alopecia comprises androgenetic alopecia and telogen alopecia;

the oleanolic acid nanoemulsion composition is used for hair care;

the oleanolic acid nanoemulsion composition is added with an excipient to be used as a dosage form for hair care, and the oleanolic acid nanoemulsion composition is used alone as a product or is mixed with other cosmetic raw materials to prepare an oleanolic acid nanoemulsion gel, an ointment, an emulsion, a spray, a lotion, a patch or a coating dosage form.