Anti-drop composition, microcapsule composition, applications and cosmetics

Abstract

The present application belongs to the field of daily chemical industry, and discloses a hair loss prevention composition, which comprises (animal) umbilical cord extract 0.1-10 wt%, Physalis alkekengi L. extract 0.1-3 wt%, and zucchini seed extract 1-5 wt%. The composition can further improve the hair loss prevention and hair growth effect; meanwhile, the formula of the present application also has the functions of soothing and repairing the scalp, and is an effective functional component with composite functions and capable of acting on the scalp. Meanwhile, the present application also discloses a microcapsule composition containing the above-mentioned composition and the application of the composition.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of new materials for daily chemicals, and particularly relates to a hair loss prevention composition, a microcapsule composition, application and a cosmetic. BACKGROUND

[0002] Normally shed hair is hair in telogen and anagen phase, because of the dynamic balance between hair in telogen and hair in anagen, the normal amount of hair can be maintained. Pathological hair loss refers to abnormal or excessive hair loss, which has many causes, including but not limited to: androgenetic alopecia, alopecia areata, endocrine alopecia, nutritional alopecia, physical alopecia, chemical alopecia, infectious alopecia, symptomatic alopecia, etc.

[0003] Most of the existing hair loss products focus on the control of hair loss factors, mainly androgenetic alopecia; androgen is converted from cholesterol by enzyme. Testosterone forms dihydrotestosterone through 5α-reductase, and the known hair loss mechanism is: 1) dihydrotestosterone binds to the androgen receptor, affecting the transcription process. 2) Dihydrotestosterone up-regulates the synthesis of TGF-β2 in the dermal papilla, TGF-β2 induces the synthesis and activation of caspase, causing apoptosis, making the telogen phase of hair advance and shortening the hair growth cycle. 3) Dihydrotestosterone binding to androgen receptor will cause molecular interference with Wnt / β-catenin, hindering the release of Wnt 10b and other beneficial hair growth factors by dermal papilla cells, and releasing interleukin 6 and other immune inflammatory factors, affecting the growth and communication of keratinocytes and fibroblasts around the hair follicle.

[0004] For example, the patent application with the subject of a compound hair growth composition for treating hair loss or alopecia and its preparation method, CN118370806A, discloses the use of ligustrum lucidum, radix falcaliae polygoni multiflori, perilla leaf, and type 17 collagen to inhibit hair loss, and its main principle is: the synergistic effect of ligustrum lucidum, radix falcaliae polygoni multiflori, and perilla leaf can significantly reduce the secretion of TGF-β1, promote the growth of new hair follicles, reduce the effect of androgens on susceptible hair follicles, and inhibit the miniaturization of hair follicles, thereby reducing hair loss.

[0005] It can be seen that the current mainstream method of preventing hair loss is to inhibit the aging of keratinocytes and fibroblasts around the hair follicle and promote the health of hair follicle cells.

[0006] The applicant's raw material supplier, Guangzhou Youtejia Biological Technology Development Co., Ltd., filed a patent application CN117100673B in 2023, with the subject of an animal umbilical cord extract for regulating hair follicle cycle and its preparation method and application, which discloses that the animal umbilical cord extract can be used to regulate hair follicle cycle, promote the entry of hair follicles in the resting phase into the growth phase, and achieve the purpose of preventing hair loss and promoting hair growth.

[0007] After the applicant experimented with the above raw materials, it was found that the above raw materials have good anti-hair loss effect. In the actual product formula research and development process, not only is it required that the product acting on the scalp has an anti-hair loss effect, but also it is hoped that it can realize maintenance of the scalp from different dimensions, further improving the application effect of the raw material. SUMMARY

[0008] Therefore, a first object of the present application is to provide an anti-hair loss composition which can further improve the anti-hair loss and hair growth effect. Meanwhile, the formula of the present application also has the functions of soothing and repairing the scalp, and is an effective functional ingredient which can act on the scalp and has composite functions.

[0009] Meanwhile, the present application also discloses a microcapsule composition containing the above composition and the application of the composition.

[0010] To achieve the first object of the present application, the following technical solutions are adopted:

[0011] An anti-hair loss composition comprises:

[0012] (animal) umbilical cord extract 0.1-10wt%;

[0013] Physalis alkekengi L. extract 0.1-3wt%;

[0014] Zucchini seed extract 1-5wt%.

[0015] The individual effects of the components of the present application are as follows:

[0016] The (animal) umbilical cord extract contains small molecular proteins, polypeptides, amino acids, vitamins and other key ingredients for regulating the physiology of the scalp and hair follicles, which is used to maintain the cell viability of keratinocytes and fibroblasts around the hair follicles and inhibit premature aging. The function of this substance has been comprehensively discussed in the patent application with the title of Animal Umbilical Cord Extract for Regulating Hair Follicle Cycle and Its Preparation Method and Application, and will not be described in detail here.

[0017] The active ingredients in the Physalis alkekengi L. extract are phytosterols, specifically β-sitosterol, stigmasterol, stigmasterol-3-O-β-D-glucoside, etc., which have the functions of reducing cholesterol content and reducing inflammation, and are also beneficial to the proliferation of fibroblasts around the hair follicles. These give the theory that phytosterols can prevent hair loss, which has the potential to prevent hair loss; at the same time, the Physalis alkekengi L. extract can be used as a plant substitute for hydrocortisone, which can inhibit phospholipase A2, soothe the skin, instantly relieve itching, reduce inflammation and allergic reactions, and repair the skin barrier.

[0018] Zucchini seed extract can up-regulate LC3A gene to activate autophagy. LC3 is the full name of MAP1LC3, which runs through the whole autophagy process and is currently recognized as an autophagy marker. LC3 of mammals can be divided into three types: LC3A, LC3B and LC3C; LC3A is modified and processed by ubiquitin-like system including Atg7 and Atg3, coupled with phosphatidylethanolamine (PE), forms LC3B and is located on the inner and outer membranes of autophagosomes. After the fusion of autophagosomes and lysosomes, LC3B on the outer membrane is cut by Atg4 to produce LC3A for recycling. Through the activation of cellular autophagy, the degradation of lysosomes, the selective removal of self-damaged, aged or excessive biological macromolecules and organelles, and the release of free small molecules for normal dynamic life processes of cells, it is particularly important for the health of hair follicle related cells. In addition, in the present application, the cell repair function of zucchini seed extract makes zucchini seed extract the main component of the formula of the present application to realize the repair function of the scalp.

[0019] Through the above mechanism analysis, it can be seen that, taking the hair follicle cycle regulation function of (animal) umbilical cord extract as the core, by further improving the proliferation level of fibroblasts around the hair follicle, improving the repair ability of the scalp cells, further increasing the number of hair follicles in the resting phase entering the growth phase, and improving the hair follicle anchoring ability.

[0020] In the further research of the present application, it is found that by reasonably adjusting the amount of each component, 5α-reductase and hyaluronidase can be effectively inhibited. These two indicators are important indicators for oil control and soothing, and are important functional parameters for scalp health. In addition, the formula of the present application has excellent free radical scavenging ability.

[0021] The formula of the present application is a composite functional material with the core function of preventing hair loss, and the functions of soothing and repairing the scalp.

[0022] It should be noted that the above-mentioned raw materials are commercial raw materials, and the amount used is the amount of commercial raw materials. The protection scope of the present application should be based on the total amount of effective components corresponding to the commercial raw materials;

[0023] In the present application, the content of effective components in (animal) umbilical cord extract is 0.22wt%; the content of effective components in Physalis alkekengi L. extract is 0.0005wt%; and the content of effective components in zucchini seed extract is 0.49wt%.

[0024] In the above anti-hair loss composition, it comprises:

[0025] (animal) umbilical cord extract 4.5-5.5wt%;

[0026] Physalis alkekengi L. extract 0.2-1wt%;

[0027] Zucchini seed extract 1.5-2.5 wt%.

[0028] In the above-mentioned anti-hair loss composition, comprising:

[0029] (animal) umbilical cord extract 5 wt.%;

[0030] Physalis alkekengi extract 0.5 wt.%;

[0031] Zucchini seed extract 2 wt.%;

[0032] Meanwhile, the present application also discloses a kind of microcapsule composition, including solvent, phospholipid, as any described above in the anti-hair loss composition;

[0033] The microcapsule in the microcapsule composition is liposome;The shell material of the liposome includes the phospholipid;

[0034] The weight ratio of the anti-hair loss composition and lecithin is 1.6-18:1-8.

[0035] As known, liposome microcapsule can be coated with oil-soluble active ingredients and water-soluble active ingredients based on its bilayer structure, which is relatively close to the skin structure and can be well compatible with the skin.

[0036] The present application uses liposome as the specific implementation form of microcapsule, which can improve the uniformity and affinity of the anti-hair loss composition on the scalp, and realize the best play of the composition effect.

[0037] As a further improvement of the present application, the present application does not use cholesterols as the mobility regulator of the liposome shell material, but uses at least part of the plant-derived sterols in Physalis alkekengi extract as the mobility regulator of the liposome shell material, which makes the liposome and the scalp better affinity, improves the coating stability of Physalis alkekengi extract in the liposome and the release effect on the scalp, and improves the proliferation promotion ability to fibroblasts around hair follicles. From the human experiment effect verification, it has better anti-hair loss effect compared with other plant-derived sterols, and has more significant anti-hair loss effect compared with liposome using cholesterols.

[0038] In the above-mentioned microcapsule composition, further comprising a solvent, the solvent is a polyol, water.

[0039] In the above-mentioned microcapsule composition, the polyol is one or a combination of butanediol, glycerol and 1,2-hexanediol.

[0040] Specifically, in the microcapsule composition of the present application, the content of water is 38-55 wt.%;

[0041] The content of polyol is 40-50 wt.%;

[0042] The phospholipid is hydrogenated lecithin and / or lecithin.

[0043] In the microcapsule composition described above, the shell material of the liposome further comprises part or all of the plant-derived sterol in the extract of Physalis alkekengi L.

[0044] In the microcapsule composition described above, the microcapsule composition contains 1.6-18 wt% of the anti-falling composition, 1-8 wt% of the phospholipid.

[0045] In the microcapsule composition described above, β-cyclodextrin is further included, and the amount of the β-cyclodextrin is 3-7 wt%.

[0046] The main function of the β-cyclodextrin is to stabilize the oil-soluble ingredient, form a complex structure of cyclodextrin-Physalis alkekengi L. extract-liposome, and more specifically, form a complex structure of cyclodextrin-oil-soluble ingredient-liposome.

[0047] The β-cyclodextrin has a more obvious effect when used in a large amount of Physalis alkekengi L. extract, and if the amount of the Physalis alkekengi L. extract is small, it is not recommended to use the β-cyclodextrin. The β-cyclodextrin has a ring structure, and its interior is a hydrophobic cavity that can accommodate the oil-soluble ingredient. When the oil-soluble ingredient is loaded and covered by the liposome, the β-cyclodextrin can play a slow-release and targeting function. Therefore, when the amount of the Physalis alkekengi L. extract is less than 0.5 wt%, it is not recommended to use the β-cyclodextrin, so as to avoid damaging the fluidity of the shell material of the liposome.

[0048] In addition, the application also discloses a use of the microcapsule composition described above in the preparation of a cosmetic.

[0049] In the use described above, the cosmetic is a cosmetic for the scalp and / or hair.

[0050] Finally, the application also discloses a cosmetic containing the microcapsule composition described above.

[0051] In the cosmetic described above, the cosmetic contains 1-20 wt% of the microcapsule composition.

[0052] In the cosmetic described above, the cosmetic is a scalp essence, a shampoo or a hair conditioner.

[0053] The application has the following beneficial effects:

[0054] The present application takes the hair follicle cycle regulation function of the (animal) umbilical cord extract as the core, further improves the proliferation level of fibroblasts around the hair follicle, improves the repair ability of scalp cells, further increases the number of hair follicles in the growth phase from the resting phase, and improves the hair retention ability of the hair follicle. By reasonably adjusting the amount of each component, 5α-reductase and hyaluronidase can be effectively inhibited. These two indicators are important indicators for oil control and soothing, and are an important functional parameter for scalp health. In addition, the formula of the present application has excellent free radical scavenging ability.

[0055] In summary, the formula of the present application is a composite functional material with the core function of preventing hair loss, and the functions of soothing and repairing the scalp. BRIEF DESCRIPTION OF DRAWINGS

[0056] Figure 1 is the experimental procedure flow chart of the hyaluronidase inhibition test;

[0057] Figure 2 is the experimental procedure flow chart of the ABTS antioxidant effect test;

[0058] Figure 3 is the test result photo of the cell autophagy test of the microcapsule composition of Example 2. DETAILED DESCRIPTION

[0059] The technical solutions of the present application will be further described below through specific embodiments. Those skilled in the art should understand that the embodiments are only to help understand the present application, and should not be regarded as specific limitations on the present application.

[0060] The specific experimental procedures or conditions are not specified in the embodiments, and can be performed according to the conventional experimental procedures described in the literature or the operation or conditions. The reagents or instruments used are not specified by the manufacturer, and are conventional reagent products that can be obtained by purchase.

[0061] The supplier information and / or product information are as follows:

[0062] The (animal) umbilical cord extract is purchased from Guangzhou Youtaijia Biotechnology Development Co., Ltd.; the effective component is mesenchymal stem cells; the content of the effective component is 5.0wt%;

[0063] The extract of Physalis alkekengi L. is purchased from Zhuhai Jiayan Biotechnology Co., Ltd.; the effective component is phytosterol; the content of the effective component is 1.0wt%;

[0064] The extract of Cucurbita pepo L. is purchased from Zhuhai Jiayan Biotechnology Co., Ltd.; the effective component is free amino acid; the content of the effective component is 5.0wt%;

[0065] Camellia seed extract, purchased from Zhuhai Jiayan Biotechnology Co., Ltd.; effective ingredient: phytosterol; content of effective ingredient: 1.0wt%;

[0066] Sunflower seed extract, purchased from Zhuhai Jiayan Biotechnology Co., Ltd.; effective ingredient: phytosterol; content of effective ingredient: 1.0wt%.

[0067] Preparation of the first part of the microcapsule composition

[0068] The formula of the microcapsule composition of the embodiments and comparative examples of the present application can refer to Table 1 and Table 2; the preparation method is as follows:

[0069] A. Take (animal) umbilical cord extract, zucchini seed extract (or camellia seed extract, sunflower seed extract), Physalis alkekengi extract, hydrogenated lecithin, water, and polyhydric alcohol according to the following mass percentage;

[0070] B. Mix and stir the Physalis alkekengi extract and hydrogenated lecithin weighed in step A, heat to 40-60℃, and stir until completely dissolved as the oil phase;

[0071] C. Mix and stir the (animal) umbilical cord extract, zucchini seed extract, water, and polyhydric alcohol weighed in step A until completely dissolved, and mix uniformly with β-cyclodextrin;

[0072] D. Mix the oil phase obtained in step B and the obtained in step C, add the obtained mixture into the reaction kettle of the supercritical device, first punch in CO2 through the CO2 high-pressure pump, adjust the temperature in the reaction kettle of the supercritical device to 30-60℃, set the pressure to 8-16Mpa, incubate under this supercritical state for 20-60min, release CO2 by pressure relief, and cool to room temperature to obtain the microcapsule composition.

[0073] Table 1 Formula table of examples

[0074] Example 1 Example 2 Example 3 Example 4 Example 5 (Animal) umbilical cord extract 0.1 5 10 5 5 Wedelia trilobata extract 0.5 0.5 3 3 0.5 Zucchini seed extract 1 2 5 2 5 Hydrogenated lecithin 5 5 8 1 5 Beta-cyclodextrin 5 5 3 7 5 Butylene glycol 40 40 40 40 40 Glycerol 3 3 3 3 3 1,2-Hexanediol 0.5 0.5 0.5 0.5 0.5 Water to 100 to 100 to 100 to 100 to 100

[0075] Table 2 Formula table of comparative examples

[0076]

[0077]

[0078] Function verification of the second part of the microcapsule composition

[0079] Test Example 1

[0080] Safety test of the microcapsule composition of Examples 1-5

[0081] The microcapsule composition of Examples 1-5 was subjected to human skin closed patch test for safety evaluation.

[0082] The human skin closed patch test was carried out according to the test requirements of the closed patch in the 2015 edition of the "Cosmetic Safety Technology Standard", and the microcapsule compositions of Examples 1-5 were tested; the negative control was pure water, and the total number of subjects was 33, with an average age of 36.39±6.09 years, and the test results are shown in Table 3.

[0083] Table 3 Patch test of microcapsule compositions of Examples 1-5 Different skin reaction score grades

[0084]

[0085]

[0086] The patch test was carried out by 33 subjects using the above microcapsule compositions, and the results showed that after the patch was removed, all the subjects were negative.

[0087] Test Example 2

[0088] 5a-reductase inhibition test of microcapsule compositions of Examples 1-5, Comparative Examples 1-7

[0089] Principle of experiment: 5a-reductase is a membrane protein enzyme dependent on reduced coenzyme II (NADPH), which is an important androgen metabolism enzyme in the skin, and can irreversibly convert testosterone (T) into dihydrotestosterone (DHT). DHT can induce excessive secretion of sebum from sebaceous glands. By inhibiting the activity of 5a-reductase to reduce the level of DHT, it can effectively alleviate the excessive secretion of sebum from sebaceous glands.

[0090] Test steps

[0091] In a 10 mL centrifuge tube, add PBS, testosterone solution, different concentrations of sample solution or control solution, enzyme extract solution, and NADPH solution in sequence, mix well, incubate at 37°C, detect the absorbance change value at 340 nm within 10 min using a UV-visible spectrophotometer, analyze and process the data to calculate the inhibition rate of the test sample on 5a-reductase.

[0092] Calculation method

[0093] 5a-reductase inhibition rate I (%) = [(A-B) / (A-C)]x100%

[0094] Where: A is the absorbance change value of the enzyme normal reaction group (containing PBS, containing testosterone solution, containing enzyme extract solution, containing NADPH solution)

[0095] B is the absorbance change value of the experimental group (containing PBS, containing testosterone solution, containing sample solution (sample group) / finasteride solution (positive control group) / water (negative control group), containing enzyme extract, containing NADPH solution)

[0096] C is the absorbance change value of the blank group (containing PBS, containing testosterone solution, containing NADPH solution)

[0097] The test results are as follows in Table 4;

[0098] Table 4 5a-reductase inhibition rate statistical results table

[0099]

[0100] Result analysis:

[0101] 1. As can be seen from Examples 1, 2 to 5, the 5a-reductase inhibition effect of Example 1 is lower than that of Examples 2 to 5, and the reason is that the effective component is low;

[0102] 2. As can be seen from Examples 2 to 5, the 5a-reductase inhibition rate of Example 2 is the best, and the possible reason is that the components in Example 2 have a synergistic effect, and the active ingredient in Examples 3 to 5 increases the use amount, and the synergistic effect between the components decreases.

[0103] 3. As can be seen from the test results of Example 2, Comparative Example 1, Comparative Example 2, Comparative Example 5, Comparative Example 6, and Comparative Example 7, when other phytosterol extracts (camellia seed extract, sunflower seed extract) are used to replace the extract of Physalis alkekengi L., the effect is obviously weaker than that of Example 2; of course, it cannot be denied that the extract of Physalis alkekengi L. is obviously better than the other two in terms of 5a-reductase inhibition rate, but the effect of Example 2 is improved by a much larger margin than the effect brought by the extract of Physalis alkekengi L. alone, therefore, we believe that the extract of Physalis alkekengi L. has synergy with other components such as (animal) umbilical cord extract and zucchini seed extract in the present formula, and at the same time, it has stronger functionality in participating in the shell material of liposomes.

[0104] 4. As can be seen from Example 2, Comparative Example 3, and Comparative Example 4, the extract of Physalis alkekengi L. and the zucchini seed extract are indispensable in the formula of the present application, and together they improve the 5a-reductase inhibition rate.

[0105] Test Example 3

[0106] Hyaluronidase inhibition test

[0107] Experimental principle: Hyaluronic acid is the most abundant component in the extracellular matrix, regulates the secretion of cytokines, and affects cell adhesion, growth, proliferation and differentiation, thus playing an important role in maintaining skin moisture and elasticity, wound healing and blood vessel formation. Hyaluronidase is a specific hyaluronidase that is strongly related to the release of histamine from mast cells and is involved in allergic reactions. Inhibition of hyaluronidase activity can ensure normal hyaluronic acid content and function. Therefore, the inhibition rate of hyaluronidase is used as an index to evaluate the soothing effect of cosmetics.

[0108] Hyaluronidase can hydrolyze sodium hyaluronate to generate N-acetylglucosamine, which can be condensed with p-dimethylaminobenzaldehyde after heating under alkaline conditions to produce a color reaction. The absorbance can be measured at 585 nm on a spectrophotometer. Samples with hyaluronidase activity inhibition can reduce the generation of chromogenic substances, thereby reducing the absorbance. According to the change in absorbance, the inhibition of hyaluronidase activity of the sample is evaluated.

[0109] Test procedure reference Figure 1 ;

[0110] The test results are shown in Table 5 below.

[0111] Table 5 Hyaluronidase inhibition rate results statistical table

[0112]

[0113] Result analysis:

[0114] 1. As can be seen from Table 5, Examples 1 to 5, the hyaluronidase inhibition rate of each sample can reach more than 60%, among which Examples 3 and 4 have the best effect, indicating that the extract of Cosmos bipinnata is the main soothing ingredient in the composition of the present application. The hyaluronidase inhibition rate of Example 5 can reach 79%, which is significantly improved compared with Example 2, indicating that the zucchini seed extract plays a secondary role in the inhibition of hyaluronic acid.

[0115] 2. As can be seen from Table 5, Comparative Examples 5 to 7, the extract of Cosmos bipinnata, Camellia seed extract and sunflower seed extract all have a certain effect on the inhibition of hyaluronic acid. As can be seen from Comparative Examples 1, 2 and Example 2, the composition of the present application has a slight advantage in soothing effect. In Comparative Example 3, the amount of Cosmos bipinnata extract is increased to 2.5 g, and the soothing effect is significantly enhanced. The soothing effect of Comparative Example 4 is weaker than that of Comparative Example 3, which further proves that the extract of Cosmos bipinnata plays a major role in soothing, and the zucchini seed extract plays a secondary role in the inhibition of hyaluronic acid.

[0116] Test Example 4

[0117] Example 1-5, Comparative Example 1-7, microcapsule compositions were tested for ABTS antioxidant effect

[0118] Principle of the experiment: ABTS method is often used to detect the in vitro antioxidant capacity of a substance.

[0119] Stable radical ABTS+solution appears blue-green color, with maximum absorption at 734 nm; this radical can be generated by the reaction of ABTS with K2S2O8. When the radical is scavenged, the number decreases, and its solution color becomes lighter, and the absorbance at 734 nm decreases, thus the ability of the sample to scavenge ABTS+can be determined.

[0120] Test procedure reference Figure 2 ;

[0121] The test results are shown in Table 6 below.

[0122] Table 6 Radical scavenging rate statistical results

[0123]

[0124]

[0125] Result analysis:

[0126] 1. As can be seen from Table 6, Examples 1 to 5, each group can exhibit excellent free radical scavenging effect, and the difference between each group is not significant;

[0127] 2. As can be seen from Table 6, Comparative Examples 1 to 4, the free radical scavenging effect of each group is between 67% and 79%, and the difference is not significant;

[0128] 3. As can be seen from Table 6, Comparative Examples 5 to 7, the free radical scavenging effect of each group is poor, and the difference is not significant;

[0129] The above cases show that the free radical scavenging effect of the present application is based on the synergistic effect of the components.

[0130] Test Example 5

[0131] Example 2 Microcapsule composition anti-shedding test (This experiment is sent to a third-party detection, and the detection unit is: Hangzhou Huan Te Biological Technology Co., Ltd.)

[0132] Principle of the experiment:

[0133] Hair loss, also known as alopecia, is a skin disease characterized by hair loss, which affects appearance, and is closely related to the number, proliferation, degeneration and apoptosis of human hair papilla cells. Human dermal papilla cells (HDPCs) play a core regulatory role in hair growth and hair follicle cycle. They are mesenchymal cells in the hair follicle, have the characteristics of adult stem cells, and can affect the development of hair follicles, the growth cycle of hair, and the regeneration of hair follicles. Papillary cells secrete various growth factors and cell signaling molecules to regulate the proliferation, differentiation and apoptosis of hair follicle epithelial cells, thereby affecting hair growth and shedding. In terms of anti-hair loss efficacy, promoting the proliferation of hair papilla cells is one of the key factors. Clinical evidence shows that the accumulation of dihydrotestosterone (DHT) in HDPCs is related to androgenetic alopecia, and 5α-reductase in the male testis converts testosterone (T) in the follicle to DHT. DHT regulates male reproductive development, testis formation, skeletal muscle growth and hair growth by activating androgen receptor (AR). However, the affinity of DHT for AR is 10 times that of T, leading to its overactivation, stimulating HDPCs to secrete TGF-β, and converting the hair follicle from the anagen phase to the catagen phase, shortening the anagen phase and inhibiting hair follicle cell proliferation. The nutritional needs of hair growth are supplied by capillaries around the hair follicle, and VEGF plays an important role in the formation of capillaries around the hair follicle. VEGF is a highly specific vascular endothelial cell growth factor that promotes angiogenesis, increases vascular permeability, promotes extracellular matrix degeneration, vascular endothelial cell migration, proliferation, and other functions. Overexpression of VEGF in outer root sheath keratinocytes can induce the formation of blood vessels around the hair follicle, leading to accelerated hair regeneration after hair loss, increased size of hair follicles and hair shafts, and increased hair density. The Wnt / β-catenin pathway has a positive effect on the growth and cycle of mammalian hair follicles. After activation, β-catenin accumulates in the cytoplasm and then translocates to the nucleus, where it interacts with Lef / Tcf transcription factors to regulate the expression of genes responsible for hair follicle growth. DHT can inhibit the Wnt / β-catenin pathway in human dermal papilla cells. DKK1 (Dickkopf-1) is a secreted glycoprotein belonging to the DKK family, which antagonizes Wnt / β-catenin signaling pathway activity by competitively binding LRP5 / 6 receptors with Wnt proteins, and regulates cell proliferation and differentiation, and is a typical inhibitor of the Wnt / β-catenin signaling pathway.

[0134] Therefore, in this experiment, human dermal papilla cells were used to construct an experimental model, and the promotion of cell proliferation and the relative expression of AR, TGF-β, DDK1, VEGF and β-catenin genes after the sample was applied to the cells were detected to determine whether the sample had anti-hair loss efficacy.

[0135] Experimental procedure:

[0136] I. Cell proliferation

[0137] 1. Cells were seeded in 96-well plates (5x103cells / well) and incubated at 37℃, 5% CO2for 24h.

[0138] 2. Drug administration: sample groups were added with fresh medium containing corresponding concentrations of samples, normal control group was replaced with fresh medium, blank group was added with blank medium, and incubated at 37℃, 5% CO2for 48h.

[0139] 3. After incubation, MTT solution was added to each well, and incubated for 4h.

[0140] 4. The medium was removed, DMSO solution was added, shaken and mixed, and the absorbance value at 490nm was measured.

[0141]

[0142]

[0143] OD490: absorbance value at 490nm

[0144] II. Relative expression of AR, TGF-β, DDK1, VEGF, β-catenin genes

[0145] 1. Cells were seeded in 6-well plates (3x105cells / well) and incubated at 37℃, 5% CO2for 24h.

[0146] 2. Drug administration: after incubation, normal control group was added with fresh complete medium, model control group was added with fresh complete medium containing (30μM DHT), sample group was added with fresh complete medium containing (30μM DHT + sample), positive control group was added with fresh complete medium containing (30μM DHT + 400μM minoxidil), and incubated at 37℃, 5% CO2for 24h.

[0147] 3. Cells were collected, total RNA was extracted from each experimental group, cDNA was synthesized, and q-PCR was used to detect the gene expression of β-actin and target genes.

[0148] 4. β-actin was used as the internal reference for gene expression, and the RNA relative expression of the target gene was calculated.

[0149] RNA relative expression = 2 ΔΔC(t)

[0150]

[0151] ΔC(t) = C(t) 目的基因 -C(t)​β-actin

[0152] The test results are shown in Table 7 below:

[0153] Table 7 AR, TGF-β, DDK1, VEGF, β-catenin gene relative expression detection results

[0154]

[0155] Summary: The cell survival rate of the sample Example 2 microcapsule composition group was significantly increased compared with the normal control group; the relative expression amounts of AR, TGF-β, and DDK1 genes were significantly reduced compared with the model control group, and the relative expression amounts of VEGF and β-catenin genes were significantly increased compared with the model control group, revealing that the sample has the effect of preventing hair loss.

[0156] Test Example 6

[0157] Autophagy test of the microcapsule composition of Example 2 (this experiment was sent to a third party for detection, and the detection unit was Hangzhou Huan Te Biological Technology Co., Ltd.)

[0158] Experimental principle:

[0159] Autophagy is a self-protection mechanism widely existing in eukaryotic cells. Cells have aging and damaged organelles, incorrectly synthesized proteins, invading bacteria, and macromolecular substances, etc. Autophagy encapsulates these substances harmful to cells in autophagosomes, and combines with lysosomes to form autolysosomes to decompose and remove these substances, maintain the homeostasis of the cell environment, and recycle the substances in the cell. The expression amount of autophagy-related protein LC3II can be detected by Western blot to determine whether the autophagy level is increased.

[0160] Test steps:

[0161] 1. Inoculate cells in a 6-well plate (3x105 cells / well), and incubate at 37°C, 5% CO2 for 24 h.

[0162] 2. After incubation, remove the culture medium, add fresh culture medium containing the sample to the sample group, replace the fresh culture medium to the normal control group, and add fresh culture medium containing 50 nM rapamycin to the positive control group, and continue to incubate at 37°C, 5% CO2 for 24 h.

[0163] 3. After incubation, lyse the cells with RIPA lysis buffer to extract proteins.

[0164] 4. Detect the LC3II protein by Western Blot method.

[0165] The test results are shown in Table 8 and Figure 3 Table 8.

[0166] Table 8 Statistics of cell autophagy test results

[0167]

[0168] Summary: Under the experimental conditions, the sample of Example 2 microcapsule composition has the effect of promoting cell autophagy.

[0169] The fourth part is the preparation of the scalp serum and the human hair loss prevention test

[0170] The formula of the scalp serum is shown in Table 9, and the preparation method is as follows:

[0171] Step 1: uniformly stir A phase in a clean and dry container, and add B phase for standby;

[0172] Step 2: start homogenization, put C phase into the pot, stop homogenization, stir to warm up to 80-85°C, and keep warm for 30 minutes;

[0173] Step 3: start the temperature reducing device, the temperature is reduced to 40-45°C, add A and D phases, and stir until transparent and clear;

[0174] Step 4: take the intermediate product to detect pH, if the pH value is higher than the standard range (pH is 5-7), add E phase;

[0175] Step 5: after uniform stirring, the scalp serum product is obtained.

[0176] Table 9 Formula of scalp serum

[0177]

[0178] Eight scalp serum samples are obtained by the formula in Table 9, and the corresponding relationship between their numbers and the used microcapsule compositions is shown in Table 10;

[0179] Table 10 Information table of scalp serum

[0180]

[0181]

[0182] Test Example 7

[0183] Example 2, Comparative Example 1 Microcapsule Composition Scalp Serum Human Test

[0184] Experimental steps:

[0185] 1. Materials and methods

[0186] 1.1. Test products: scalp serum 1 (Example 2), scalp serum 2 (Comparative Example 1).

[0187] 1.2. Control product: the corresponding test product base formula product without anti-hair loss efficacy ingredients, tested in parallel with the test product.

[0188] 1.3. Wash-off period product: same as the control product, both the test group and the control group use the control product during the wash-off period.

[0189] 1.4. Subjects: a total of 90 effective subjects completed the test, aged 24 to 53. Among them, the test group was divided into 2 groups, each group had 30 people, and the total number of 2 groups was 26 men and 34 women, the male to female ratio was adjusted to be balanced in each experimental group; the control group had 30 people, 12 men and 18 women. The subjects met the selection and exclusion criteria.

[0190] 1.5 Test method: the test was conducted in accordance with the specific requirements of the "Cosmetic Safety Technical Specification" (2015 edition). The subjects were recruited according to the selection and exclusion criteria, and the qualified subjects were subjected to a 2-week wash-off period. After the end of the wash-off period, the subjects were subjected to 60 times of combing method again, and only those with more than 10 hairs counted as hair loss could enter the formal test. The subjects were divided into test product group and control product group according to the stratified random method, and were given test products and control products respectively. The products were used correctly according to the product instructions for 12 weeks, and the subjects were required to record the use diary and strictly follow the restrictions during the test period. Hair loss counting was performed at each visit before product use (baseline value) and 4 weeks, 8 weeks, and 12 weeks after product use, and the results were recorded. At the same time, before use, 2 weeks, and 4 weeks, the subjects were tested for scalp physiological parameters, stratum corneum moisture content, transdermal water loss, oil content, scalp red area, and elasticity value.

[0191] The test results can be seen in Table 11;

[0192] Table 11 Human experiment test results

[0193]

[0194]

[0195] Results analysis:

[0196] Through the continuous use of the product by the subjects for 84 days, it is known that the scalp stratum corneum moisture content of the test group essence 1 is improved by 59.88% at the 4-week return visit, the transdermal water loss is improved by 25.05%, the oil content is improved by 25.07%, and the scalp elasticity value is improved by 13.82%; the hair loss count of the test group essence 1 is improved by 59.06% at the 12-week return visit, and the local hair density is increased by 4.56%; it is shown that the test product essence 1 has the effects of soothing, repairing the scalp, and preventing hair loss for the hair loss population. The test group essence 2 has an improved effect compared with the control group, but the improvement effect is not ideal, which shows that the extract of Physalis alkekengi var. franchetii plays a strengthening effect in preventing hair loss.

[0197] In summary, the microcapsule composition of the anti-hair loss composition of the present application has the effects of preventing hair loss and promoting hair growth, soothing, and repairing the scalp barrier.

[0198] The embodiments presented herein are only selected from combinations of all possible embodiments. The appended claims should not be limited by the embodiments of the present application. Some numerical ranges used in the claims include sub-ranges within them, and variations in these ranges should also be covered by the appended claims.

Claims

1. A hair loss prevention composition, characterized in that, include: Animal umbilical cord extract 4.5–5.5 wt%; 0.2–1 wt% of *Heliotropium indicum* extract; Zucchini seed extract 1.5–2.5 wt.

2. A microcapsule composition, characterized in that, The product includes solvents, phospholipids, and an anti-hair loss composition; the anti-hair loss composition includes animal umbilical cord extract, *Heliotropium indicum* extract, and zucchini seed extract. The microcapsules in the microcapsule composition are liposomes; the shell material of the liposomes includes the phospholipids. The microcapsule composition comprises: Animal umbilical cord extract 4.5–5.5 wt%; 0.2–1 wt% of *Heliotropium indicum* extract; Zucchini seed extract 1.5–2.5 wt%; Phospholipids 1-8 wt%.

3. The microcapsule composition according to claim 2, characterized in that, The solvent is a polyol or water; the polyol is one or more combinations of butanediol, glycerol, and 1,2-hexanediol; the phospholipid is hydrogenated lecithin and / or lecithin.

4. The microcapsule composition according to claim 2, characterized in that, It also includes β-cyclodextrin, wherein the amount of β-cyclodextrin used is 3 to 7 wt%.

5. Use of the microcapsule composition according to any one of claims 2 to 4 to prepare a cosmetic, wherein the cosmetic is a cosmetic applied to the scalp and / or hair, and the cosmetic has an anti-hair loss effect.

6. A cosmetic product, characterized in that, The cosmetic contains the microcapsule composition as described in any one of claims 2 to 4, and is a cosmetic with anti-hair loss effects.

7. The cosmetic product according to claim 6, characterized in that, The cosmetic contains 1-20 wt% of a microcapsule composition.

8. The cosmetic product according to claim 6, characterized in that, The cosmetic product is a scalp serum, shampoo, or conditioner.

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