Application of ellagic acid nanoparticles in preparation of food, health food, medicine, cosmetics and daily chemical products with efficacy of preventing or treating alopecia

Abstract

The invention provides ellagic acid nanoparticles which can solve the problems of extremely low solubility, transdermal absorption performance and oral bioavailability of ellagic acid without adding a large number of auxiliary materials and a complex preparation process. When being used for preparing food, health-care food, medicines, cosmetics and daily chemical products with the effect of preventing or treating alopecia, the compound can improve hair growth of an alopecia model mouse, the external treatment effect is optimal, the hair of the mouse is thick and glossy, and the hair weight and length are remarkably improved compared with those of a model group; the oral administration group also has a better treatment effect; meanwhile, modeling after pre-gavage shows an excellent hair generation condition compared with a model group, so that the traditional Chinese medicine composition has a prevention effect at the same time. Meanwhile, the ellagic acid nanoparticle foaming agent has excellent dispersity and permeability, and the light foam structure of the ellagic acid nanoparticle foaming agent is easier to diffuse in the skin in the transverse direction and the longitudinal direction. Furthermore, the ellagic acid nanoparticles have the effect of penetrating into the deep layer of the skin, and can reach and target the fascia layer of the skin at the deepest.

Description

Technical Field

[0001] The invention belongs to the field of food raw materials, external preparation raw materials, cosmetic raw materials, and pharmaceutical raw materials, and specifically relates to the application of ellagic acid nanoparticles in the preparation of food, health food, medicine, cosmetics, and daily chemical products with the efficacy of preventing or treating hair loss. Technical Background

[0002] Hair loss refers to the phenomenon that hair falls off from the scalp under the influence of various factors. Its causes are complex, and genetic factors, endocrine disorders, nutritional deficiencies, mental stress, diseases and drugs may all cause hair loss. Hair loss is mainly divided into androgenic alopecia, alopecia areata, growth effluvium, telogen effluvium, traction alopecia, scarring alopecia, postpartum alopecia, nutritional metabolic alopecia, and infectious alopecia. Androgenic alopecia (AGA), also known as male pattern baldness and seborrheic alopecia, is a common type of hair loss. Androgenic alopecia usually starts in adolescence or post-adolescence as a non-scarring, progressive hair follicle miniaturization hair loss disease. It is mainly manifested as hair loss starting from the bilateral frontal angles and gradually extending to the top of the head. It can also be manifested as a receding hairline on the forehead with or without progressive hair reduction and thinning on the top of the head. 50% of men will experience AGA at the age of 50, and the probability of occurrence increases to 70% with age.

[0003] Ellagic acid (EA), also known as retrogallic acid, is a plant polyphenol with CAS number 476-66-4 and molecular formula C 14 H6O8, molecular weight 302.19Da, melting point>360℃; characteristic wavelengths of ultraviolet absorption are 254nm and 366nm, insoluble in ethers, slightly soluble in water, alcohol, dimethyl sulfoxide, soluble in pyridine and alkaline solutions. It is widely found in various fruits, nuts and other plants, such as grapes, pomegranates, apples, cherries, raspberries, strawberries, blueberries, tara, walnuts, and Euryale ferox, and has a wide range of effects, such as anti-inflammatory, antibacterial, and antiviral, especially anti-oxidation, anti-aging, whitening, and astringent. Ellagic acid has been included in the International Standard Catalog of Cosmetic Ingredients (INCI) internationally, and China has also included it in the International Standard Chinese Name Catalog of Cosmetic Ingredients (2015 edition). Ellagic acid has been applied to many types of cosmetics such as whitening and anti-aging due to its high safety, mild effect on the skin, and significant efficacy. Functional cosmetics made with ellagic acid as raw material have great research and development value and market prospects.

[0004] Ellagic acid belongs to Class IV of the Biopharmaceutical Classification System (BCS), i.e., low solubility and low permeability drugs. High lattice force (melting point 360°C) is the root cause of the poor solubility of ellagic acid. The application of EA is seriously hindered by its extremely low solubility (<10μg / mL in water). In the previous study, the inventor used ultrasonic packaging to make ellagic acid self-assemble to form nanoparticles (EANPs), which significantly improved the solubility of EA (>120 times). As a result, two Chinese invention patents were applied for and one academic paper was published, hereinafter referred to as "Patent 1", "Patent 2", and "Article 1":

[0005] Patent 1: Gu Wei; Cheng Jianming; Wang Yiwei; Qi Shuyang; Ji Jing. A nanosuspension of ellagic acid with high solubility and antioxidant activity and its application in oral, topical preparations and cosmetics [P]. Jiangsu Province: CN202211465349.3, 20221122);

[0006] Patent 2: Gu Wei; Chen Jun; Yang Kangli; Qian Yuerong; Tong Huangjin; Hu Qinglian; Li Feng; Zhang Shunan; Xu Fei; Zhao Hongyu. A nanosuspension of ellagic acid and its application in oral, topical preparations and cosmetics [P]. Jiangsu Province: CN202111461097.2.;

[0007] Article 1: Wei Gu, Ruolin Kong, Shuyang Qi, Xuxi Cheng, Xuyi Cai, Ziyun Zhou, Shunan Zhang, Hongyu Zhao, Jinyun Song, Qinglian Hu, Huiwen Yu, Huangjin Tong*, Yiwei Wang*, Tulin Lu*. Sono-assembly of ellagic acid into nanostructures significantly enhances aqueous solubility and bioavailability[J]. Food Chemistry. 2024, 442: 138485. doi: 10.1016 / j.foodchem.2024.138485. PubMed PMID: 38278106. Summary of the invention

[0008] The invention provides an application of ellagic acid nanoparticles in preparing food, health food, medicine, cosmetics and daily chemical products with the efficacy of preventing or treating hair loss, wherein the ellagic acid nanoparticles are prepared by the following method: ellagic acid powder is added to a preparation solvent and prepared by ultrasonic process; the preparation solvent is pure water or an organic solvent or a mixed solution of an organic solvent and water, and the organic solvent is selected from one or more of ethanol, propylene glycol, butylene glycol, pentanediol and glycerol; the ultrasonic process conditions are: ellagic acid feed concentration ≥1.0 mg / mL, ultrasonic power ≥100 w, ultrasonic frequency ≥20 kHz, ultrasonic temperature ≥30 DEG C, and ultrasonic time ≥5 min; the obtained suspension is subjected to centrifugation, filtration, sedimentation and other methods to remove solid part, and the supernatant is taken to obtain the ellagic acid nanoparticles.

[0009] As an improvement, in the ultrasonic process conditions, the ultrasonic temperature is ≥ 35°C, more preferably 45°C.

[0010] As another improvement, the ellagic acid feed concentration in the ultrasonic process condition is ≥2.5 mg / mL, more preferably 20.0 mg / mL.

[0011] As another improvement, the preparation solvent is water, or a mixed solution of an organic solvent and water, wherein the ratio of the organic solvent to water is preferably 0:1-3:1 by volume.

[0012] As an improvement, the medicine, cosmetic, and daily chemical product are tinctures, gels, ointments, creams, patches, plasters, foams, lotions, films, and patches, and foams are most preferred.

[0013] As a further improvement, the medicine, cosmetics, and daily chemical products are foaming agents, and their preparation method is as follows: ellagic acid nanoparticles are prepared into a dispersion using an ultrasonic disperser; a surfactant is weighed and added to the dispersion, and a moisturizer and a stabilizer are added at the same time, and the mixture is completely and evenly mixed; triethanolamine is added dropwise to neutralize the system until the pH value reaches 6.5-7.0, so as to form a stable drug-containing liquid foam matrix; the prepared drug-containing liquid foam matrix is ​​loaded into a foam pump bottle, and dispersed by a foam pump to form a stable foam, so as to obtain an ellagic acid nanoparticle foaming agent.

[0014] As another improvement, the food, health food, and medicine are in the form of oral liquid dosage form, lyophilized powder, granules, tablets, capsules, effervescent tablets, or soft capsules.

[0015] As a further improvement, the hair loss is androgenic alopecia, alopecia areata, anagen effluvium, telogen effluvium, traction alopecia, cicatricial alopecia, postpartum alopecia, nutritional metabolic alopecia, infectious alopecia, and androgenic alopecia is preferred.

[0016] Beneficial Effects

[0017] The present invention provides a kind of ellagic acid nanoparticles, which can solve the problems of extremely low solubility, transdermal absorption performance and oral bioavailability of ellagic acid without adding a large amount of auxiliary materials and without going through a complicated preparation process. It is used in the preparation of food, health food, medicine, cosmetics and daily chemical products with the effect of preventing or treating hair loss, and the hair growth of hair loss model mice can be improved, and the external treatment effect is the best. The hair of mice is dense and shiny, and both the weight and length of hair are significantly improved compared with the model group; the oral group also has a better therapeutic effect. The ellagic acid nanoparticles are first gavaged into mice for 7 days, and then the formation of hair loss is induced by applying testosterone propionate every day. It can be seen that compared with the model group, the hair generation situation is significantly improved, and it can be known that it has a preventive effect. The ellagic acid nanoparticles show extremely significant efficacy in treating or preventing hair loss, which is not only related to the significant enhancement of its solubility, but also related to its own nano-nized properties, showing the development prospects of ellagic acid nanoparticles in the field of external preparations and the field of oral preparations. At the same time, the ellagic acid nanoparticle foam showed a wide diffusion range and a faster diffusion speed, indicating that the foam has better dispersibility and permeability, and its light foam structure is easier to diffuse horizontally and vertically in the skin. Furthermore, the ellagic acid nanoparticles showed the effect of penetrating deep into the skin, reaching and targeting the skin fascia layer, which may be an important reason why the ellagic acid nanoparticles have excellent efficacy in preventing / treating hair loss after external use. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The present invention will be further described below in conjunction with the accompanying drawings:

[0019] Figure 1 :The cumulative transdermal amount of ellagic acid in different test solutions changes with time in the in vitro transdermal experiment. * P<0.5.

[0020] Figure 2 : Therapeutic / preventive effects of drugs on hair loss model mice (n=6).

[0021] Figure 3 : Hair length and hair weight, compared with the model group, * P<0.5, ** P < 0.1; # P<0.5, ## P<0.1; ns: no significance, no significant difference.

[0022] Figure 4 :EANPs foaming agent.

[0023] Figure 5 : Agar penetration experiment.

[0024] Figure 6 : Skin fluorescence microscopy and skin HE staining microscopy. DETAILED DESCRIPTION

[0025] The technical solution of the present invention is further described below in conjunction with specific examples, which is not a limitation of the present technical solution. Diols such as ethanol, propylene glycol, butylene glycol, pentanediol, and glycerol are commonly used solvents, moisturizers, penetration enhancers, or preservatives in topical preparations or oral preparations. The most commonly used 1,3-propylene glycol, 1,3-butylene glycol, and 1,2-pentanediol are selected for testing in the following examples, but are not used as limitations of the present technical solution. In addition, the "ellagic acid nanoparticles" in this patent include "ellagic acid nanosuspension" in "patent 1", "ellagic acid nanosuspension" in "patent 2", and "Ellagic Acid Nanostructures" in article 1.

[0026] Example 1 Preparation of Ellagic Acid Nanoparticles (EANPs)

[0027] Ellagic acid (EA) powder is added to the preparation solvent and prepared by ultrasonic process; the ultrasonic process conditions are: ellagic acid feed concentration ≥ 1.0 mg / mL, ultrasonic power ≥ 100w, ultrasonic frequency ≥ 20kHz, the obtained suspension is centrifuged, filtered, sedimented and other methods to remove the solid part, and the supernatant is taken to obtain EANPs. The preparation method can be optimized from the following aspects: the preparation solvent is pure water or an organic solvent or a mixed solution of an organic solvent and water, preferably water, the organic solvent is preferably one or more of ethanol, propylene glycol, butylene glycol, pentanediol, and glycerol, and the ratio of organic solvent to water in the mixed solution of organic solvent and water is preferably 0: 1-3: 1; the ultrasonic temperature in the ultrasonic process conditions is ≥ 30°C, more preferably ≥ 35°C, and most preferably 45°C; the ultrasonic time in the ultrasonic process conditions is ≥ 5min; the ellagic acid feed concentration is preferably ≥ 2.5 mg / mL, and most preferably 20.0 mg / mL.

[0028] Example 2 Determination of ellagic acid concentration by ultraviolet spectrophotometry (refer to Example 3 in "Patent 2")

[0029] Establishment of standard curve: accurately weigh 2.5 mg of ellagic acid standard in a 50 mL volumetric flask and dilute with methanol to prepare a 50 μg / mL standard solution for standby use. Then dilute with ethanol to obtain 12.5, 10, 6.25, 5, 3.125, 1.5625, 0.78125 μg / mL ellagic acid standard solutions, measure the absorbance value under ultraviolet wavelength of 254 nm, draw a standard curve with the mass concentration of ellagic acid standard solution as the abscissa (X) and the corresponding absorbance value (Y) as the ordinate, and perform linear regression on the standard curve to obtain the regression equation: Y = 0.0933X-0.0027, correlation coefficient R2 =0.9999, indicating that ellagic acid has a good linear relationship in the range of 0.78125-12.5 μg / mL.

[0030] Example 3 Preparation and Characterization of EANPs

[0031] Preparation of EA NPs: Take an appropriate amount of ellagic acid, prepare it into a 20 mg / mL suspension with ultrapure water, shake and mix, place it in an ultrasonic cleaner and ultrasonicate it at 45°C for 1 hour, then use an ultrasonic cell disruptor to ultrasonicate it for 2 minutes, place the probe 3 cm below the liquid in an ice water bath, the condition is 50% power (900W), and the ultrasound is turned on for 2 seconds and off for 2 seconds. After the ultrasound is completed, place it in a 50mL centrifuge tube, centrifuge it at 4000rpm for 16 minutes, and take the supernatant to obtain the EA NPs suspension. Freeze-dry it for 24 hours to obtain the EA NPs freeze-dried powder.

[0032] Characterization of EA NPs: ① Investigation of EA concentration: The concentration of ellagic acid was determined by the method in Example 2, and the EA content in the prepared EA NPs was 86.96±1.12%. ② Investigation of the equilibrium solubility of EA NPs in water: 20.0 mg EA and EA NPs freeze-dried powder were added to a 10 mL centrifuge tube, and then 4 mL ultrapure water was added. After sealing with a sealing film, it was placed in a 37°C constant temperature water bath for 24 hours, and 3 copies were paralleled. Stop shaking, take the supernatant after free sedimentation for 48 hours, and immediately add methanol to dilute it to the linear range of the standard curve after separation. The absorbance at 254 nm was measured by an ultraviolet spectrophotometer, and the total concentration of EA was substituted into the standard curve to calculate the equilibrium solubility. The results are shown in Table 1. The equilibrium solubility of EA NPs after acoustic cavitation treatment reached 1.64 mg / mL, which was significantly higher than that of free EA.

[0033] Table 1 EA content in EANPs and the equilibrium solubility of EA and EANPs in water (n=3)

[0034]

[0035] Note: Compared with EA, *P<0.05, **P<0.01, ***P<0.001; ND means “not detected”.

[0036] ③ Determination of particle size, potential, and PDI After EA NPs were reconstituted and diluted with ultrapure water, the particle size, PDI, and ζ-potential of the EA NPs suspension were measured using a particle size analyzer. According to Table 2, the particle size of the nanoparticles in the EA NPs suspension is about 270 nm, the PDI is about 0.17, and the particle size is relatively uniform. The ζ potential is -31.6±2.9 mV, which has good stability.

[0037] Table 2 Particle size, PDI, ζ-potential measurement results (n=3)

[0038]

[0039] Example 4 Preparation of free EA solution with the same concentration

[0040] The corresponding free EA solution was prepared using 50% ethanol + 25% propylene glycol (common tincture preparation method) as the solvent, with the ellagic acid concentration (1.64 mg / mL) in Example 3 as the standard. The concentration of ellagic acid in the solution in Example 3 and Example 4 was further verified by UV measurement, as shown in Table 3.

[0041] Table 3 Ellagic acid content in each group

[0042]

[0043] Note: The EA contents in the table are all data obtained by testing using the equilibrium solubility determination method in Example 3.

[0044] Example 5 Preparation of EA saturated aqueous solution (EA is in free state at this time)

[0045] The EA saturated aqueous solution was prepared as shown in Table 3.

[0046] Example 6 In vitro transdermal test

[0047] HPLC method for determination of ellagic acid content (refer to Example 11 in "Patent 2"): ① Chromatographic conditions: The chromatographic column was Hedera C18 column (250 mm × 4.6 mm, 5 μm), and the mobile phase was acetonitrile-0.2% phosphoric acid aqueous solution (V 乙腈 :V 磷酸 =20:80), flow rate 1.0mL / min, detection wavelength 254nm, column temperature 30℃, injection volume 10μL, spectrum acquisition time 15min. ②Linear relationship investigation Take the reference solution to make a 100μg / mL mother solution, dilute it with methanol to 100, 50, 25, 12.5, 6.25, 3.125, 1.5625, 0.78125, 0.390625, 0.1953125μg / mL, inject it into HPLC, and use the mass concentration of ellagic acid reference solution as the horizontal axis (X) and the corresponding peak area as the vertical axis to establish a standard curve (Y), which is Y=1.5937X+0.4033, R 2 =0.9992, and the linear range is 0.1953125~100μg / mL.

[0048] The abdominal skin of rats was used as the transdermal barrier in the in vitro transdermal experiment. Before the experiment, the abdominal skin of healthy rats was cut, cleaned with saline to remove surface residues, and then cut into appropriate sizes to ensure that the skin was intact. The skin stratum corneum was fixed upward between the test cell and the receiving cell of the Franz diffusion cell. The test solution was added to the test cell. The test solution was divided into three groups: ellagic acid nanoparticles, free ellagic acid solution with the same ellagic acid concentration as the nanoparticles, and ellagic acid saturated aqueous solution. The volume of each group was 1 mL to ensure that the reagent covered the entire skin surface. 10 mL of pH 7.4 phosphate buffer was added to the receiving cell as the receiving solution, and its temperature was maintained at 37 ± 0.5 ° C. At the same time, a magnetic stirrer was used to continuously stir to maintain the uniformity of the solution and the stability of mass transfer. The experiment was divided into multiple time points to collect receiving solution samples, including 0, 4, 8, 12, 16, 20 and 24 hours. After each 200 μL sample was collected, an equal amount of fresh phosphate buffer was immediately added. The samples were collected and the concentration of ellagic acid was determined by high performance liquid chromatography (HPLC) to calculate the cumulative amount of ellagic acid that penetrated the skin and entered the receptor fluid per unit time. Figure 1 It can be seen that the transdermal performance of EANPs is significantly better than that of the EA saturated aqueous solution group (Example 5); and when the solvent system is changed to make the EA concentration (Example 4) equivalent to the EANPs aqueous solution (Example 3), the transdermal absorption performance is also equivalent.

[0049] Example 7 Study on the efficacy of topical anti-hair loss drugs

[0050] A C57BL / 6J mouse hair loss model was used, and the drug groups were shown in the table. Testosterone propionate was applied in the morning every day to induce the formation of a hair loss model, and topical drugs were applied in the afternoon. The three groups listed in Table 3 were prepared into gels according to the method in Example 8 and applied for administration, wherein the concentration of ellagic acid was 5%, and the blank group was applied with blank gel; in order to investigate the effect of oral administration, an oral gavage group of EANPs prepared in Example 3 was added; the mice were photographed and the hair growth on the back was observed. Figure 2The experimental results showed that the blank group mice had normal hair growth throughout the experiment, and the skin was gradually covered with hair, while the model group hair growth was significantly inhibited, and the back skin was smooth and hairless, verifying the successful establishment of the model. The positive control group used commercially available minoxidil tincture (minoxidil content 5%, solvent system 50% ethanol + 25% propylene glycol), and hair growth was significantly improved after 21 days of administration. The therapeutic effects after topical administration of each ellagic acid group are ranked as follows: Example 3>> Example 4> Example 5. It can be seen that the EANPs group has dense and shiny hair, showing a very significant efficacy in treating hair loss, which is not only related to the significantly enhanced solubility of EANPs (Example 3> Example 5), but also related to its own nano-nized properties (Example 3> Example 4); At the same time, oral administration of EANPs also has a better therapeutic effect, although it is much lower than the topical group (Example 3 oral group). EANPs were first gavaged for 7 days, and then testosterone propionate was applied every day to induce hair loss. It can be seen that compared with the model group, the hair growth was significantly improved (prevention group in Example 3), which shows its preventive effect. At the same time, on the 21st day, the hair growing on the back of the mice was completely removed and packed in marked plastic bags. The hair weight of each group was measured and recorded, and 30 hairs were randomly taken out from each group and the length was measured using ImageJ software. The results are shown in Figure 3 The results show that among the drug administration groups, the order of hair weight and length is Example 3 topical group > prevention group > Example 3 oral group > Example 4 > Example 5, among which the topical effect of Example 3 is significantly better than the positive control minoxidil group. In summary, ellagic acid nanoparticles are effective in the treatment and prevention of hair loss, especially when used as topical preparations.

[0051] Example 8 Preparation of gel

[0052] First, weigh carbomer 940 and add it to an appropriate amount of pure water, slowly add it while stirring until it is completely swollen to form a uniform basic solution, and let it stand overnight to ensure sufficient swelling; add glycerol and methylparaben to the swollen carbomer basic solution, mix it on a magnetic stirrer to form a preliminary gel matrix, and slowly drop triethanolamine to neutralize it until the pH value reaches 6.5-7.0, so that the gel forms a stable and transparent state. The drug is dispersed with pure water, treated with an ultrasonic disperser (ultrasonic power 50W, ultrasonic time 2 minutes), a stable and uniform dispersion is obtained, and it is slowly added to the prepared gel matrix, and a homogenizer is used to obtain a drug gel after sufficient stirring. The obtained gel has stable properties, a pH value in the range of 6.5-7.0, good smearing performance and penetration-promoting effect, and can be applied to topical treatments in hair loss areas.

[0053] Example 9 Preparation of Foaming Agent

[0054] The drug was dispersed with pure water and treated with an ultrasonic disperser (ultrasonic power 50W, ultrasonic time 2 minutes) to prepare a uniform dispersion. Then, a certain amount of surfactant (such as polysorbate-80) was weighed and added to the dispersion, and a moisturizer (such as glycerol) and a stabilizer (such as carbomer 940) were added to improve the stability of the foam. The mixed solution was placed on a magnetic stirrer and stirred until it was completely uniform. Subsequently, triethanolamine was added dropwise to neutralize the system until the pH value reached 6.5-7.0 to form a stable basic liquid foam matrix. Next, the prepared liquid matrix was loaded into a foam pump bottle and dispersed by a foam pump to form a stable foam, and finally a drug-containing foam was obtained. The obtained foam has a light texture and fine and uniform foam. It can be used for topical treatment of hair loss areas. It is easy to apply and has an excellent user experience. EANPs foams such as Figure 4 shown.

[0055] Example 10 Agar penetration experiment

[0056] The agar permeation experiment aims to evaluate the diffusion ability of EANPs gel and foam in agar matrix to verify the superiority of their permeability performance. The experimental process is as follows: First, prepare a 2% agar matrix solution, add agar powder to boiling water, stir until completely dissolved, and then pour into 2cm 2 mold, and wait for it to cool and solidify to form a uniform agar layer. In the center of the agar layer, use a hole puncher to make a small hole with a diameter of about 6 mm, add 50 μL of rhodamine B solution, and use a micropipette to add EANPs gel and EANPs foam to the small holes of different culture dishes, respectively, adding the same volume (about 100 μL) to each hole. Place the agar gel in a constant temperature incubator and maintain a constant temperature of 37°C to simulate the body temperature environment. During the incubation period, record the diameter of the diffusion circle of each group in the agar matrix every 0.5, 1, 2, and 4 hours, and use a digital camera to record the diffusion image. The experimental results are as follows: Figure 5 As shown in the figure, the diffusion of EANPs gels of different concentrations in agar shows that with the increase of ellagic acid concentration, the color depth and area of ​​the diffusion circle are enhanced. From the concentration range of 0.2% to 5%, the gels with higher concentrations (such as 2% and 5%) showed a more obvious diffusion trend at each time point (0.5, 1, 2, and 4h) after the experiment. This shows that concentration is an important factor affecting transdermal diffusion ability. In addition, the effect of time on the diffusion depth is also very significant. As time goes on, the diffusion circle gradually deepens and expands, especially after 2 and 4h, the diffusion range of each group is significantly improved.

[0057] Compared with gels, EANPs foams exhibited excellent lateral transdermal diffusion effects, diffusing laterally on the surface of the agar block at all concentrations with good uniformity. At the same time, it showed concentration dependence. At the same time point (such as 1 and 2h), the diffusion range of higher concentrations (2% and 5%) of foams was significantly larger than that of low concentration groups (such as 0.2% and 0.5%). Compared with gels, the diffusion circle of foams was more uniform in color and had clearer boundaries, especially at early time points (such as 0.5 and 1h). High-concentration foams showed faster diffusion rates, which may be related to the thinner structure and superior adhesion of foams. Time extension also further enhanced the diffusion capacity of foams, but the diffusion effect of the low-concentration group gradually stabilized, while the high-concentration group still maintained significant growth.

[0058] Comparing the two dosage forms, the diffusion range of foams at different concentrations is generally larger than that of gels, and the diffusion speed is faster. This may be because the foam has better dispersibility and permeability on the agar surface, and its light foam structure is easier to diffuse into the agar matrix, while the gel may cause some obstacles to diffusion due to its higher viscosity. In addition, at higher concentrations (such as 5%), the diffusion advantage of the foam is more significant, and its light foam structure is easier to diffuse horizontally and vertically in the skin, indicating that the foam has greater potential as an external preparation of EANPs.

[0059] Example 11 Study on the skin distribution of ellagic acid nanoparticles

[0060] Male SD rats were taken, their abdominal hair was removed after anesthesia, and they were killed by cervical dislocation. The intact abdominal skin was cut off, the subcutaneous fat tissue was removed, and the rats were washed in physiological saline. The rat skin stratum corneum was fixed upward in a transdermal diffusion cell (effective diffusion area 3.14 cm 2 ), 8 mL PBS (pH 7.4) was added to the receiving cell as the receiving solution, 1 mL of each group of ellagic acid (Example 5) and ellagic acid nanoparticle solution (Example 3) was added to the supply chamber, and the mixture was stirred at 37°C and 400 r·min -1 The skin was then magnetically stirred at a speed of 1000 nm and protected from light for 12 hours. After the treatment, the skin was removed and washed with warm water to remove the residual preparation on the surface. The skin at the effective administration site was cut and fixed in a cryoembedding medium. Frozen sections were prepared and the drug distribution in the skin was observed under a fluorescence microscope.

[0061] The results of fluorescence microscopy and HE staining microscopy of the skin after administration of ellagic acid and ellagic acid nanoparticles are shown in Figure 6, it can be seen that 12 hours after administration, from the intradermal distribution, the fluorescence of the skin treated with ellagic acid is mainly distributed in the outermost stratum corneum and hair follicles, while the fluorescence of the skin treated with ellagic acid nanoparticles is not only distributed in the stratum corneum and hair follicles, but also observed to be distributed in the deep fascia layer of the skin. It can be seen that after ellagic acid is prepared as nanoparticles, it can be delivered deep into the skin, as deep as the fascia layer. In summary, ellagic acid nanoparticles have a strong affinity with the skin, reduce the barrier function of the skin, and can penetrate deep into the skin, reaching and targeting the deepest fascia layer of the skin, indicating the application potential of ellagic acid nanoparticles in the preparation of topical preparations with non-invasive deep skin drug delivery function, which may be an important reason for the excellent prevention / treatment of hair loss efficacy of ellagic acid nanoparticles after topical application.

[0062] Example 12 Preparation of topical preparations for treating / preventing hair loss using ellagic acid nanoparticles

[0063] The obtained ellagic acid nanoparticles are prepared by the method of any one of Examples 1 or 3 or Claims 1 to 6, and used as raw materials in the preparation of external preparations, including medicines, cosmetics, and daily chemical products, including the following dosage forms: tinctures, gels, ointments, creams, patches, plasters, foams, lotions, films, and films, and the most preferred foams have the effect of preventing or treating hair loss, and the deep penetration of the drug is conducive to the efficacy of these preparations. The hair loss is androgenic alopecia, alopecia areata, growth effluvium, telogen effluvium, traction alopecia, scarring alopecia, postpartum alopecia, nutritional metabolic alopecia, and infectious alopecia, preferably androgenic alopecia.

[0064] Example 13 Preparation of ellagic acid nanoparticles for oral preparation for treating / preventing hair loss

[0065] The obtained ellagic acid nanoparticles are prepared by the method of any one of Example 1 or 3 or Claims 1 to 6, and used as raw materials in the preparation of oral preparations, including foods, health foods, and medicines, including the following dosage forms: oral liquid dosage forms, lyophilized powders, granules, tablets, capsules, effervescent or soft capsules, which have the effect of preventing or treating hair loss, and the improvement of drug solubility and oral bioavailability are conducive to the efficacy of these preparations. The hair loss is androgenic alopecia, alopecia areata, anagen effluvium, telogen effluvium, traction alopecia, cicatricial alopecia, postpartum alopecia, nutritional metabolic alopecia, infectious alopecia, preferably androgenic alopecia.

Claims

1. Application of ellagic acid nanoparticles in the preparation of food, health food, medicine, cosmetics, and daily chemical products for preventing or treating hair loss, characterized in that: The ellagic acid nanoparticles are prepared by the following method: ellagic acid powder is added to a preparation solvent and prepared by ultrasonic process; the preparation solvent is pure water or an organic solvent or a mixed solution of an organic solvent and water, and the organic solvent is selected from one or more of ethanol, propylene glycol, butylene glycol, pentanediol, and glycerol; the ultrasonic process conditions are: ellagic acid feed concentration ≥1.0 mg / mL, ultrasonic power ≥100w, ultrasonic frequency ≥20kHz, ultrasonic temperature ≥30°C, and ultrasonic time ≥5min; after ultrasonication, the suspension is subjected to physical method to remove the solid part, and the physical method includes centrifugation, filtration, and sedimentation, and the supernatant is taken to obtain the ellagic acid nanoparticles.

2. The use according to claim 1, characterized in that: In the ultrasonic process conditions, the ultrasonic temperature is ≥35°C.

3. The use according to claim 2, characterized in that: The ultrasonic temperature in the ultrasonic process condition is 45°C.

4. The use according to claim 1, characterized in that: The ellagic acid feed concentration in the ultrasonic process conditions is ≥2.5 mg / mL.

5. The use according to claim 4, characterized in that: The concentration of ellagic acid in the ultrasonic process is 20 mg / mL.

6. The use according to claim 1, characterized in that: The preparation solvent is water, or a mixed solution of an organic solvent and water, wherein the volume ratio of the organic solvent to water is 0:1-3:

1.

7. The use according to any one of claims 1 to 6, characterized in that: The medicines, cosmetics and daily chemicals are tinctures, gels, ointments, creams, patches, plasters, foams, lotions, films and patches.

8. The use according to claim 7, characterized in that: The medicine, cosmetics and daily chemical products are foaming agents, and the preparation method thereof is as follows: preparing a dispersion liquid by using an ultrasonic disperser with the ellagic acid nanoparticles in any one of claims 1 to 7; weighing a surfactant and adding it to the dispersion liquid, and adding a moisturizer and a stabilizer at the same time, and mixing them completely and evenly; dripping triethanolamine to neutralize the system until the pH value reaches 6.5-7.0, so as to form a stable drug-containing liquid foam matrix; loading the prepared drug-containing liquid foam matrix into a foam pump bottle, dispersing it by a foam pump to form stable foam, and obtaining the ellagic acid nanoparticle foaming agent.

9. The use according to any one of claims 1 to 6, characterized in that: The hair loss is androgenic alopecia, alopecia areata, anagen effluvium, telogen effluvium, traction alopecia, cicatricial alopecia, postpartum alopecia, nutritional metabolic alopecia, and infectious alopecia.

10. The use according to claim 9, characterized in that: The hair loss is androgenic alopecia.

Images