Polypeptide composition containing cacumen biotae extract and application thereof

Abstract

The invention discloses a polypeptide composition containing a cacumen biotae extract and application thereof, and relates to the technical field of cosmetics, the polypeptide composition comprises the cacumen biotae extract and polypeptide, and the polypeptide comprises tripeptide-1, palmitoyl tripeptide-1 and acetyl tetrapeptide-3. The composition disclosed by the invention can effectively promote the synthesis of collagen, is beneficial to supplementing scalp nutrition, can inhibit the expression of acetyl-CoA carboxylase and the synthesis or secretion of skin grease, and has the effects of preventing hair loss, growing hair and controlling oil.

Description

Technical Field

[0001] The present disclosure relates to the technical field of cosmetics, and particularly relates to a polypeptide composition containing Platycladus orientalis extract and its application. Background Art

[0002] Hair loss refers to the abnormal shedding of human hair, which is manifested as hair loss or thinning in some scalp areas. Hair loss affects 20% of men in the 20-year-old age group and is increasing at a rate of 10% per 10 years. In other words, nearly 50% of men around 50 years old are deeply troubled by hair loss. Most hair loss is caused by male hormones and is related to the 5α-lidarutase enzyme present in the hair root sheath. The 5α-lidarutase enzyme can convert the male hormone testosterone into 5α-dihydrotestosterone (5α-DHT). 5α-DHT has an inhibitory effect on hair matrix cells, activates sebaceous glands to promote sebum secretion, makes hair shiny, and prevents dryness. However, abnormal increase in sebaceous gland secretion will affect hair follicle development, resulting in smaller hair follicles and thinner hair, thus leading to massive hair loss in patches. From the physiological mechanism of hair loss, controlling abnormal sebaceous gland secretion and promoting hair follicle development are two important means to prevent hair loss.

[0003] Platycladus orientalis refers to the dry twigs and leaves of the plant Platycladus orientalis in the Cupressaceae family, which has the effects of cooling blood for hemostasis, resolving phlegm and relieving cough, promoting hair growth and blackening hair. Clinically, it is commonly used to treat hematemesis, epistaxis, hemoptysis, hematochezia, metrorrhagia and menorrhagia, cough due to lung heat, hair loss due to blood heat, and premature whitening of hair and beard. Modern medical experiments have proved that Platycladus orientalis extract can increase cell nutrition, stimulate scalp hair follicles, has a certain protective effect on follicle damage caused by cyclophosphamide, and slows down the hair entering the catagen phase; it can also induce follicle cells to express β-catenin, control follicle development and regulate cell differentiation, thereby promoting hair growth (Li Guihua et al., Discussion on the treatment of seborrheic alopecia with traditional Chinese medicine Platycladus orientalis). In addition, Chinese patent application CN105106028A discloses a polypeptide composition for hair growth, which contains polypeptides with the function of stimulating collagen production and polypeptides for promoting keratin production, and can supplement nutrition for the scalp and new hair while promoting hair growth, thereby enhancing the effect of hair growth and thickening. Chinese patent application CN111658580A discloses an essence containing multiple peptides and plant extracts, which combines polypeptides with Platycladus orientalis extract and other plant extracts, and further promotes hair growth by utilizing the synergistic effect between plant extracts and polypeptides.

[0004] Promoting hair follicle development and promoting hair growth are one of the means of preventing hair loss intervention. The above technologies only intervene in hair loss problems in terms of hair follicle development and hair growth, but do not effectively control abnormal sebaceous gland secretion, so there are still certain limitations. Summary of the Invention

[0005] The present disclosure relates to a polypeptide composition containing Platycladus orientalis leaf extract and its application. The composition not only retains the promoting effect of Platycladus orientalis leaf extract and polypeptides on hair growth, but also has the effect of reducing scalp sebum secretion after the Platycladus orientalis leaf extract is compounded with specific polypeptides, achieving the purpose of dual-effect anti-hair loss.

[0006] On the one hand, the present disclosure provides a composition comprising Platycladus orientalis leaf extract and polypeptides, wherein the polypeptides comprise Tripeptide-1, Palmitoyl Tripeptide-1 and Acetyl Tetrapeptide-3.

[0007] Tripeptide-1 is a tripeptide composed of glycine, histidine and lysine, and is a Matrikine signal peptide. It can regulate the activities of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs), and promote the synthesis of extracellular matrices such as collagen and glycosaminoglycans. It is an important regulator for skin repair and wound healing. Palmitoyl Tripeptide-1 is obtained by modifying the N-terminus of the peptide chain of Tripeptide-1 with palmitic acid, and has better transdermal performance. It can promote the skin to synthesize a large amount of collagen and polysaccharides, tighten the skin, lock water and moisturize, fade wrinkles, and deeply repair the skin. Acetyl Tetrapeptide-3 can stimulate dermal papilla or fibroblasts around the hair follicle to generate matrix proteins such as type III collagen and laminin, and type VII collagen, making the hair follicle more plump and healthy, and contributing to hair growth.

[0008] The inventors of the present disclosure unexpectedly found that a synergistic effect was produced after the Platycladus orientalis leaf extract and polypeptides were compounded. It can not only significantly promote the synthesis of collagen, but also inhibit the expression of acetyl-CoA carboxylase (ACC). Microscopic examination of the hair cross-section shows that the hair is formed by three-layer structure. The innermost part is the medulla, the middle is collagen, and the outermost layer is the cuticle. Therefore, the increase in collagen synthesis is not only beneficial to the growth and formation of hair, but also enhances the generation of collagen in the skin around the hair follicle, supplements the nutrition of new hair, strengthens the hair root, and achieves the effect of strengthening and toughening hair. ACC is a biotin enzyme isolated from the enzyme system that synthesizes long-chain fatty acids from acetyl-CoA (acetyl CoA). It catalyzes the conversion of acetyl CoA into malonyl CoA using the energy provided by ATP in vivo, and is the first rate-limiting enzyme in fatty acid synthesis, directly participating in fatty acid synthesis. Therefore, inhibiting the activity or expression of ACC can reduce the sebum storage in the sebaceous glands, thereby alleviating seborrhea. The composition provided by the present disclosure can achieve the effects of anti-hair loss and hair growth by supplementing scalp nutrition, promoting hair growth and controlling scalp sebum overflow.

[0009] The Platycladus orientalis leaf extract described in the present disclosure refers to a substance containing various active ingredients obtained from Platycladus orientalis leaves through extraction, physical pressing, or enzymatic hydrolysis. Quercitrin is one of the main active ingredients of Platycladus orientalis leaves and is generally used as an indicator of the content of active ingredients in the extract. High-content quercitrin can be obtained by various different extraction processes, which may be familiar to those skilled in the art, such as maceration, decoction, and so on. Those skilled in the art should clearly understand that quercitrin is only an indicator feature in the Platycladus orientalis leaf extract. In addition to quercitrin, the other active ingredients contained in Platycladus orientalis leaves are also included in the Platycladus orientalis leaf extract, and the present disclosure does not mean to equate the Platycladus orientalis leaf extract with quercitrin.

[0010] In some embodiments, the Platycladus orientalis leaf extract is prepared by successively extracting Platycladus orientalis leaves with a 70% ethanol solution, adsorbing with macroporous resin, and performing water precipitation.

[0011] In some embodiments, the preparation method of the Platycladus orientalis leaf extract includes the following steps:

[0012] S1. Reflux extract Platycladus orientalis leaves with a 70% ethanol solution, filter, and take the extract;

[0013] S2. Concentrate the extract to remove ethanol and then filter it. Load it onto AB-8 macroporous resin, first wash the impurities with water, and then elute with a 70% ethanol solution, and collect the eluate;

[0014] S3. Concentrate the eluate to remove ethanol, dissolve it in water, let it stand, filter, and take the filtrate;

[0015] S4. Add 1,3-propanediol and PEG-40 hydrogenated castor oil to the filtrate to obtain the Platycladus orientalis leaf extract; the volume ratio of the filtrate, 1,3-propanediol, and PEG-40 hydrogenated castor oil is 19:20:1.

[0016] Polypeptides are formed by connecting multiple amino acids with side chains through peptide bonds, and active groups are usually connected to the side chains; the Platycladus orientalis leaf extract contains a large number of active ingredients, and some active ingredients may react with the active groups on the side chains of polypeptides, resulting in polypeptide denaturation. The inventors of the present disclosure observed that when a composition obtained by compounding the Platycladus orientalis leaf extract with a polypeptide was left standing for a period of time, solids precipitated in the solution, showing a very unstable composition system. After a large amount of creative work, the inventors of the present disclosure found that the Platycladus orientalis leaf extract prepared by the above preparation method has good compatibility with polypeptides, solving the problem of poor stability after compounding the Platycladus orientalis leaf extract with polypeptides.

[0017] On the other hand, the present disclosure provides the use of the above composition in the preparation of a beauty composition for supplementing scalp nutrition, promoting hair growth, or preventing hair loss.

[0018] On the other hand, the present disclosure provides the use of the above composition in the preparation of a cosmetic composition for promoting collagen synthesis, increasing skin elasticity or improving skin firmness.

[0019] On the other hand, the present disclosure provides the use of the above composition in the preparation of a cosmetic composition for oil control, acne treatment or repair.

[0020] On the other hand, the present disclosure provides the use of the above composition in the preparation of a cosmetic composition for inhibiting the expression of acetyl-CoA carboxylase.

[0021] On the other hand, the present disclosure provides the use of the above composition in the preparation of a cosmetic composition for inhibiting skin oil synthesis or secretion.

[0022] On the other hand, the composition of the present disclosure can be incorporated as an active ingredient into related preparations to obtain cosmetics or daily chemical products for preventing hair loss, promoting hair growth or controlling oil. In some embodiments, the cosmetics or daily chemical products include shampoo, conditioner, hair care lotion, hair care essential oil, hair wash and care essence, hair oil, hair care lotion, scalp and hair nourishing product, hair essence, scalp paste, hair gel, hairspray, hair mask or eyebrow care lotion.

[0023] The present disclosure has the following advantages and effects:

[0024] 1. The composition of the present disclosure has a significant effect of promoting collagen synthesis, which can not only promote hair growth, but also enhance the generation of collagen in the skin around the hair follicles, supplement the nutrition of new hair, strengthen the hair roots, and achieve the effect of strengthening and toughening hair. In addition, by promoting collagen synthesis, the composition of the present disclosure is also beneficial to increasing skin elasticity or improving skin firmness.

[0025] 2. The composition of the present disclosure has a significant effect of inhibiting ACC expression, which can inhibit skin oil synthesis or secretion, and achieve the effect of reducing the sebum storage in sebaceous glands and alleviating sebum overflow; when the composition of the present disclosure is applied to the scalp, it can further relieve the occurrence of seborrheic alopecia by controlling scalp oil secretion, so as to achieve the effect of preventing hair loss. In addition, by improving problems such as excessive skin oiliness and skin water-oil imbalance, the composition of the present disclosure can reduce the occurrence of acne or pimples, and also contribute to the recovery of the skin after the occurrence of acne or pimples, and has the effects of oil control, acne treatment and repair.

[0026] 3. The polypeptide and the Platycladus orientalis extract in the composition of the present disclosure can form a stable compounding system, which can meet the requirements of long-term storage and use of the product. Detailed embodiments

[0027] To make the objectives, features, and advantages of the present disclosure more apparent and understandable, the present disclosure will be further described in detail below with reference to the embodiments. Obviously, the described embodiments are some, but not all, of the embodiments of the present disclosure. All other embodiments obtained by those of ordinary skill in the art based on the embodiments in the present disclosure without creative efforts fall within the scope protected by the appended claims of the present disclosure.

[0028] Preparation Example 1

[0029] A Platycladus orientalis leaf extract is prepared through the following steps:

[0030] S1. Take dry Platycladus orientalis leaves, pulverize them, add 70% ethanol, and reflux for 1 h. Filter to obtain the filtrate, and obtain an ethanol extract.

[0031] S2. Fill AB-8 macroporous resin into a chromatography column for standby. Concentrate the ethanol extract to remove ethanol, filter, pour the filtrate into the chromatography column, wash away impurities with water, and then elute with 70% ethanol. Collect the eluate.

[0032] S3. Concentrate the eluate to remove ethanol, dissolve it in water multiple times, let it stand, filter after insoluble substances precipitate, and obtain a water precipitation solution.

[0033] S4. According to the volume ratio of the water precipitation solution, 1,3-propanediol, and PEG-40 hydrogenated castor oil of 19:20:1, add 1,3-propanediol and PEG-40 hydrogenated castor oil to the water precipitation solution to obtain Platycladus orientalis leaf extract A.

[0034] Take a sample and determine the quercitrin content by HPLC. The measured quercitrin content is 1.101 mg / g.

[0035] Preparation Example 2

[0036] A Platycladus orientalis leaf extract is prepared through the following steps:

[0037] S1. Take dry Platycladus orientalis leaves, pulverize them, add 70% ethanol, and reflux for 1 h. Filter to obtain the filtrate, and obtain an ethanol extract.

[0038] S2. Fill AB-8 macroporous resin into a chromatography column for standby. Concentrate the ethanol extract to remove ethanol, filter, pour the filtrate into the chromatography column, wash away impurities with water, and then elute with 70% ethanol. Collect the eluate.

[0039] S3. Concentrate the eluate to remove ethanol, dissolve it in water multiple times, let it stand, filter after insoluble substances precipitate, and obtain a water precipitation solution.

[0040] S4. Concentrate and dry the water precipitation solution to obtain Platycladus orientalis leaf extract B.

[0041] Sampling, and determining the quercitrin content therein by HPLC. The measured quercitrin content is 59.277 mg / g.

[0042] Preparation Example 3

[0043] A Platycladus orientalis leaf extract is prepared by the following steps:

[0044] S1. Take dry Platycladus orientalis leaves, pulverize them, add 70% ethanol, and reflux for 1 h. Filter to obtain the filtrate, and obtain the ethanol extract;

[0045] S2. Fill AB-8 macroporous resin into a chromatography column for standby; concentrate the ethanol extract until ethanol-free, filter, pour the filtrate into the chromatography column, wash away impurities with water, and then elute with 70% ethanol, and collect the eluate;

[0046] S3. The eluate is separated by centrifugation, coarse filter paper, and filter paper in sequence to obtain a refined solution;

[0047] S4. Add 10% wt of β-cyclodextrin to the refined solution for inclusion, and then spray-dry to obtain Platycladus orientalis leaf extract C.

[0048] Sampling, and determining the quercitrin content therein by HPLC. The measured quercitrin content is 23.38 mg / g.

[0049] Preparation Example 4

[0050] A Platycladus orientalis leaf extract is prepared by the following steps:

[0051] S1. Take dry Platycladus orientalis leaves, pulverize them, add 70% ethanol, and reflux twice, 2 h each time. Filter to obtain the filtrate, and obtain the ethanol extract;

[0052] S2. Fill D101 macroporous resin into a chromatography column for standby; concentrate the ethanol extract until ethanol-free, filter, pour the filtrate into the chromatography column, wash away impurities with water, and then elute with 70% ethanol, and collect the eluate;

[0053] S3. Concentrate the eluate until ethanol-free, add 5% wt of maltodextrin, and then spray-dry to obtain Platycladus orientalis leaf extract D.

[0054] Preparation Example 5

[0055] An Eclipta prostrata extract is prepared by the following steps:

[0056] S1. Take dry whole Eclipta prostrata herb, pulverize it, add 75% ethanol, and reflux twice, 2 h each time. Filter to obtain the ethanol extract;

[0057] S2. Concentrate the ethanol extract until ethanol-free, add water, stir at 60 °C for one hour, and centrifuge to obtain the supernatant;

[0058] S3. Spray-dry the supernatant to obtain Eclipta prostrata extract.

[0059] Preparation Example 6

[0060] A cornel extract is prepared by the following steps:

[0061] S1. Take the dried whole herb of cornel, crush it, add purified water and reflux for extraction twice, 2 hours each time, filter until clear and transparent to obtain an aqueous extract;

[0062] S2. Concentrate the aqueous extract to an extract with a weight ratio of 1:2 to the raw materials, add 4 times the amount of 95% ethanol, stir while adding, let stand, centrifuge after 2 hours, and take the supernatant;

[0063] S3. Concentrate the supernatant to a specific gravity of 1.1 and freeze-dry to obtain the cornel extract.

[0064] Examples 1-6

[0065] The composition is prepared according to the following formula:

[0066] Mass percentage (%) Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Tripeptide-1 2 2 2 / / / Acetyl Tetrapeptide-3 1 1 1 / / / Palmitoyl Tripeptide-1 0.2 / / / / / Tripeptide-1 Copper / 0.2 / / / / Biotin Tripeptide-1 / / 0.2 / / / Platycladus orientalis Extract D / / / 0.4 / / Eclipta prostrata Extract / / / / 0.4 / Cornus officinalis Extract / / / / / 0.4 Phosphate Buffered Saline (PBS) Make up to 100 Make up to 100 Make up to 100 Make up to 100 Make up to 100 Make up to 100

[0067] The preparation steps of Examples 1-6 are as follows: Weigh the powder in the formula amount, mix them, add PBS and mix well, and filter through a 0.22 μm microporous filter membrane to obtain the composition.

[0068] Example 7

[0069] The composition is prepared by mixing the composition solutions of Example 1 and Example 4 in equal volumes.

[0070] Example 8

[0071] The composition is prepared by mixing the composition solutions of Example 1 and Example 5 in equal volumes.

[0072] Example 9

[0073] The composition is prepared by mixing the composition solutions of Example 1 and Example 6 in equal volumes.

[0074] Example 10

[0075] The composition is prepared by mixing the composition solutions of Example 2 and Example 4 in equal volumes.

[0076] Example 11

[0077] The composition is prepared by mixing the composition solutions of Example 3 and Example 4 in equal volumes.

[0078] Examples 12-14 Platycladus orientalis polypeptide composition

[0079] A Platycladus orientalis polypeptide composition is prepared according to the following formula:

[0080] Component Mass percentage Tripeptide-1 0.1％ Palmitoyl Tripeptide-1 0.01％ Acetyl Tetrapeptide-3 0.05％ Platycladus orientalis Extract Solution 20％ PEG-40 Hydrogenated Castor Oil 1％ 1,3-Propanediol 20％ 10% Citric Acid Appropriate amount Water Make up to 100%

[0081] Weigh PEG-40 hydrogenated castor oil and acetyl tetrapeptide-3 according to the formula, dissolve them in water to obtain solution A; mix tripeptide-1 and palmitoyl tripeptide-1, dissolve them in water and adjust the pH to 4.0 - 5.5 with 10% citric acid to obtain solution B; after mixing solution A and solution B, add the formula amount of Platycladus orientalis extract solution, then add 1,3-propanediol and water to obtain the composition.

[0082] Among them, the Platycladus orientalis extract solution in Example 12 is Platycladus orientalis extract A, and the Platycladus orientalis extract solutions in Examples 13 and 14 are prepared according to the following formula:

[0083]

[0084] Weigh PEG-40 hydrogenated castor oil and mix it with Platycladus orientalis extract B or Platycladus orientalis extract C, dissolve them in water, then add 1,3-propanediol, make up the water, and filter through a 0.45μm aqueous filter membrane to obtain it.

[0085] Example 15 Plant Polypeptide Anti-Hair Loss Essence

[0086] A plant polypeptide anti-hair loss essence is prepared according to the following formula:

[0087]

[0088]

[0089] Add the formula amount of disodium EDTA, sodium hyaluronate oligosaccharide and dipropylene glycol to water, heat it to 75 - 80°C while stirring, cool it to 60°C after uniform dispersion, then add panthenol, p-hydroxyacetophenone, 1,2-hexanediol, continue stirring and cool it to 40°C, add the formula amount of the composition in Example 12 and ethanol, and adjust the pH to 5 - 7 with arginine to obtain it.

[0090] Example 16 Collagen I Content Test

[0091] 16.1 Reagents and Materials

[0092] Complete medium, phosphate buffer solution (PBS), 0.25% trypsin digestion solution (prepared by dissolving 0.25 g of trypsin in 100 mL of water), RIPA lysis buffer, collagen I ELISA kit, BCA protein kit.

[0093] 16.2 Instruments

[0094] Microplate reader, CO2 incubator, laminar flow hood, air bath thermostatic shaker.

[0095] 16.3 cell line

[0096] Human skin fibroblasts (HSF).

[0097] 16.4 Test samples and grouping

[0098] 16.4.1 Test samples

[0099] Example 1, Example 2, Example 4, Example 6, Example 7, Example 9, Example 10, with a test concentration of 100 ppm.

[0100] 16.4.2 Grouping

[0101] Blank control group: Add PBS.

[0102] UV group: UV radiation, add PBS.

[0103] Experimental group: UV radiation, add test samples.

[0104] 16.5 Experimental method

[0105] Take HSF cells in the exponential growth phase with good condition, add 0.25% trypsin digestion solution, digest to make adherent cells detached, count (1 - 4)×10 5 cells / mL, and make a cell suspension.

[0106] Take an appropriate amount of cell suspension and inoculate it into a 12 - well plate, and culture it in a CO2 incubator for 24 h. Aspirate the complete medium in the wells, add an appropriate amount of PBS to wash the cells until colorless. The blank control group adds 100 μL PBS and supplements the complete medium to 1 mL without UV irradiation; the UV group and the experimental group add an appropriate amount of PBS, place it under an 80 mJ / cm 2 UV lamp and irradiate for 15 min, with a distance of 15 cm between the lamp source and the culture flask. After irradiation, discard the PBS, the UV group adds 100 μL PBS solution and complete medium to 1 mL, and the experimental group adds 100 μL test samples and complete medium to 1 mL. The blank control group, the UV group, and the experimental group continue to incubate in a 37°C, 5% CO2 incubator for 48 h.

[0107] After the culture is completed, collect the cells, centrifuge to obtain the cell pellet, add RIPA lysis buffer, homogenize with a vortex mixer 3 times (30 s / time, with an interval of 3 min), and centrifuge at 12000 rpm for 10 min. Aspirate the supernatant and detect it with an ELISA kit, and detect the total protein concentration of the supernatant with a BCA protein quantification kit.

[0108] 16.6 Experimental results

[0109] Collagen is one of the core proteins that make up hair and is also an important structural protein in the skin around hair follicles. In an environment with excessive ultraviolet exposure, the activity of collagenase increases significantly, and the synthesis of collagen is also inhibited. In this experiment, test samples were used to treat cells irradiated with ultraviolet rays, and the content of collagen I in the corresponding cells was detected to determine whether the composition of the present disclosure can promote the synthesis of collagen.

[0110] The results of the effect of the test samples on the content of collagen I are shown in Table 1.

[0111] Table 1 Results of the effect of test samples on the synthesis of collagen I by HSF

[0112] Group Relative content of Collagen I (Mean±SD) Blank control group 100.00％±7.50％ UV group <![CDATA[42.47% ± 5.17% ### > Example 1 68.40％±6.75％** Example 2 57.14％±4.10％** Example 4 22.24％±3.01％** Example 6 73.35％±10.99％** Example 7 80.98％±13.09％** Example 9 58.99％±5.19％** Example 10 49.48％±4.58％

[0113] Note: Compared with the blank control group, ### P < 0.001; compared with the UV group, **P < 0.01.

[0114] The results showed that compared with the blank control group, after UV irradiation, the relative content of collagen I in the UV group decreased significantly; compared with the UV group, the compositions of Example 1, Example 2, Example 6, Example 7, and Example 9 could significantly increase the content of collagen I, while the composition of Example 4 failed to promote the synthesis of collagen and instead led to a significant decrease in the content of collagen I, and the composition of Example 10 did not show an obvious effect on collagen I.

[0115] In particular, the results of Example 1 show that the polypeptide composition of tripeptide-1, palmitoyl tripeptide-1, and acetyl tetrapeptide-3 can promote the synthesis of collagen I. Example 4 proves that the use of Platycladus orientalis extract alone will further inhibit the synthesis of collagen I. However, Example 7 proves that when tripeptide-1, palmitoyl tripeptide-1, acetyl tetrapeptide-3, and Platycladus orientalis extract are used in combination, they show excellent effects in promoting the synthesis of collagen I, and this effect is also much better than the composition of Example 1. Replacing palmitoyl tripeptide-1 with tripeptide-1 copper gives the polypeptide composition of tripeptide-1, tripeptide-1 copper, and acetyl tetrapeptide-3 in Example 2, which also has the effect of promoting collagen synthesis. However, the results of Example 10 prove that when tripeptide-1, tripeptide-1 copper, acetyl tetrapeptide-3, and Platycladus orientalis extract are used in combination, no synergistic effect is shown, and its promoting effect on collagen synthesis is weaker than that of Example 2. Further, Example 6 proves that the use of Cornus officinalis extract alone can effectively promote the synthesis of collagen I. However, Example 9 proves that when tripeptide-1, palmitoyl tripeptide-1, acetyl tetrapeptide-3, and Cornus officinalis extract are used in combination, the promoting effect of this composition on the synthesis of collagen I not only does not increase but decreases instead. The effect of this composition in promoting the synthesis of collagen I is inferior to both the polypeptide combination of Example 1 and the effect of using Cornus officinalis extract alone, proving that there is an antagonistic effect between tripeptide-1, palmitoyl tripeptide-1, acetyl tetrapeptide-3, and Cornus officinalis extract.

[0116] The above results strongly prove that the interaction results between polypeptides and plant extracts cannot be predicted in advance. The combination of tripeptide-1, palmitoyl tripeptide-1, acetyl tetrapeptide-3, and Platycladus orientalis extract in the present disclosure has a synergistic effect, achieving unexpected technical effects. It can significantly promote collagen synthesis, which is beneficial to promoting hair growth, supplementing the nutrition of new hair, strengthening the hair roots, and achieving the effect of strong and tough hair. In addition, by promoting collagen synthesis, the composition of the present disclosure is also beneficial to increasing skin elasticity or improving skin firmness.

[0117] Example 17 ACC Content Test

[0118] 17.1 Reagents and Materials

[0119] Complete medium, phosphate buffer solution (PBS), FFA (composed of linoleic acid and palmitic acid in a molar ratio of 1:1), 0.25% trypsin digestion solution (prepared by dissolving 0.25 g of trypsin in 100 mL of water), RIPA lysis buffer, ACC ELISA kit, BCA protein kit.

[0120] 17.2 Instruments

[0121] Microplate reader, CO2 incubator, laminar flow hood, air bath thermostatic shaker.

[0122] 17.3 Cell Line

[0123] Human sebaceous gland cells (SZ-95).

[0124] 17.4 Test Samples and Grouping

[0125] 17.4.1 Test Samples

[0126] Examples 1 - 11, test concentration 10 ppm.

[0127] 17.4.2 Grouping

[0128] Blank control group: Add PBS.

[0129] Model group: Induced by FFA, add PBS.

[0130] Experimental group: Induced by FFA, add test samples.

[0131] 17.5 Experimental Method

[0132] Take a bottle of SZ-95 cells in the exponential growth phase with good condition, add 0.25% trypsin digestion solution, digest to make the adherent cells detached, count (1 - 4)×10 5 cells / mL to prepare a cell suspension.

[0133] Take an appropriate amount of cell suspension and inoculate it on a 12-well plate containing complete medium, and culture it in a constant temperature CO2 incubator for 24 h. Except for the blank control group adding an equal amount of PBS, add FFA to each of the remaining wells to make the final concentration 225 μmol / mL. While inducing the model, the model group adds an equal amount of PBS, and the experimental group adds the corresponding test samples respectively, and culture it in a constant temperature CO2 incubator for 48 h.

[0134] After the culture is completed, collect the cells, centrifuge to obtain the cell precipitate, add RIPA lysis buffer, homogenize it 3 times with a vortex mixer (30 s / time, interval 3 min), and centrifuge at 12000 rpm for 10 min. Aspirate the supernatant for detection with an ELISA kit, and detect the total protein concentration of the supernatant with a BCA protein quantification kit.

[0135] 17.6 Experimental Results

[0136] ACC is a biotin enzyme isolated from the enzyme system for synthesizing long-chain fatty acids from acetyl-CoA, and it catalyzes the conversion of acetyl coenzyme A to malonyl coenzyme A by using the energy provided by ATP in vivo. As the first rate-limiting enzyme in fatty acid synthesis, ACC is directly involved in fatty acid synthesis. Inhibiting the activity or expression of ACC can reduce the sebum storage in sebaceous glands, thereby alleviating sebum overflow. In this experiment, SZ-95 cells induced by FFA were treated with test samples, and the ACC content in the corresponding cells was detected to determine whether the disclosed composition can inhibit the expression of ACC.

[0137] The results of the effect of test samples on ACC expression are shown in Table 2.

[0138] Table 2 Results of the effect of test samples on the expression of ACC in SZ-95

[0139] Group Relative content of ACC (Mean±SD) Blank control group 100.00％±9.92％ Model group <![CDATA[130.40% ± 4.35% ## > Example 1 162.42％±26.48％ Example 2 160.66％±20.90％* Example 3 150.81％±15.06％ Example 4 137.78％±19.21％ Example 5 119.34％±22.94％ Example 6 135.74％±9.65％ Example 7 103.53％±15.32％* Example 8 140.28％±13.18％ Example 9 176.12％±10.54％*** Example 10 143.66％±5.27％* Example 11 137.09％±7.33％

[0140] Note: Compared with the blank control group, ## P < 0.01; compared with the model group, ***P < 0.001, *P < 0.05.

[0141] The results showed that compared with the blank control group, after induction by FFA, the content of ACC in the model group increased significantly; compared with the model group, the polypeptide combination of tripeptide-1, palmitoyl tripeptide-1 and acetyl tetrapeptide-3 (Example 1) did not show a statistically significant inhibitory or promoting effect on ACC expression; the extract of Platycladus orientalis (Example 4), the extract of Eclipta prostrata (Example 5) and the extract of Cornus officinalis (Example 6) also did not show a statistically significant inhibitory or promoting effect on ACC expression. However, the combination of tripeptide-1, palmitoyl tripeptide-1, acetyl tetrapeptide-3 and the extract of Platycladus orientalis (Example 7) could significantly reduce the content of ACC; after replacing the extract of Platycladus orientalis with the extract of Eclipta prostrata, the composition (Example 8) did not show an inhibitory or promoting effect on ACC expression; after replacing the extract of Platycladus orientalis with the extract of Cornus officinalis, the composition (Example 9) even showed an effect of increasing the content of ACC.

[0142] On the other hand, after replacing palmitoyl tripeptide-1 in the composition of Example 7 with tripeptide-1 copper, the composition (Example 10) also showed an effect of increasing the content of ACC; after replacing palmitoyl tripeptide-1 in the composition of Example 7 with biotin tripeptide-1, the composition (Example 11) did not show a statistically significant inhibitory or promoting effect on ACC expression.

[0143] The above results prove that Tripeptide-1, Palmitoyl Tripeptide-1, Acetyl Tetrapeptide-3 and Platycladus orientalis extract are a special combination. Only this combination shows the effect of reducing the content of ACC, can significantly inhibit the expression of ACC, reduce the sebum storage in sebaceous glands, relieve excessive sebum overflow, and has an oil control effect. Replacing one of the polypeptides or the plant extract component does not have the effect of inhibiting the expression of ACC, and may even promote the expression of ACC, further exacerbating skin oil secretion.

[0144] Example 18 Oil Content Test

[0145] 18.1 Reagents and Materials

[0146] 0.25% Trypsin Digest (prepared by dissolving 0.25 g of trypsin in 100 mL of water), PBS, FFA (prepared by mixing linoleic acid and palmitic acid in a molar ratio of 1:1), Oil Red O Staining Kit.

[0147] 18.2 Instruments

[0148] CO2 Incubator, Laminar Flow Hood, Air Bath Thermostatic Shaker.

[0149] 18.3 Cell Line

[0150] Human Sebaceous Gland Cells SZ-95.

[0151] 18.4 Test Samples and Grouping

[0152] 18.4.1 Test Samples

[0153] Composition of Example 7, test concentrations are 10 ppm and 100 ppm;

[0154] Simvastatin, test concentration is 0.05 μM.

[0155] 18.4.2 Grouping

[0156] Experimental Group: Test Samples, FFA;

[0157] Normal Group: PBS;

[0158] Model Group: PBS, FFA.

[0159] 18.5 Experimental Method

[0160] Take a bottle of SZ-95 cells in the exponential growth phase with good condition, add 0.25% trypsin digest, digest to make the adherent cells detached, and count (1 - 4) × 10 5Cells were resuspended at a density of [[X]] cells / mL to prepare a cell suspension. An appropriate amount of the cell suspension was inoculated onto a 12-well plate containing complete medium and cultured in a constant-temperature CO2 incubator for 24 h. Except for the normal group which was added with PBS, FFA was added to each of the other wells to a final concentration of 225 μmol / mL. While inducing the establishment of the model, the corresponding test samples were added to the experimental groups and cultured in a constant-temperature CO2 incubator for 48 h. The culture medium was aspirated and discarded, and staining was performed according to the instructions of the Oil Red O staining kit.

[0161] The solution in the well plate was aspirated and discarded, isopropanol was added to dissolve the Oil Red O staining solution, and the absorbance value was measured at 492 nm using an enzyme-linked immunosorbent assay (ELISA) reader. The relative content of cellular lipid secretion was calculated based on the absorbance value.

[0162] 18.6 Experimental results

[0163] Oil Red O is a fat-soluble dye that can be highly dissolved in fat. Its staining principle is that Oil Red O can specifically adsorb to neutral triglycerides, lipids, and lipoproteins in tissues and cells, thereby staining the fat. FFA is an inducer. Under the stimulation and induction of FFA, SZ-95 cells will secrete a large amount of lipids that can be stained by Oil Red O. In this experiment, SZ-95 cells induced by FFA were treated with test samples, and the amount of lipid production in SZ-95 cells was detected to determine whether the test samples could inhibit the lipid secretion of human sebaceous gland cells.

[0164] The results of the effect of the test samples on the lipid secretion of SZ-95 cells are shown in Table 3.

[0165] Table 3 Relative lipid secretion of SZ-95 cells

[0166]

[0167] Note: Compared with the normal group, ## P < 0.01; compared with the model group, **P < 0.01, *P < 0.05.

[0168] The experimental results showed that the composition of Example 7 could significantly inhibit the synthesis and secretion of lipids by sebaceous gland cells, slow down lipid deposition, and could be used to improve problems such as excessive skin oiliness and skin water-oil imbalance. When the composition of Example 7 was applied to the scalp, it could further relieve the occurrence of seborrheic alopecia by controlling scalp lipid secretion, thereby achieving the effect of preventing hair loss; it could also reduce the occurrence of acne or pimples and contribute to the recovery of the skin after the occurrence of acne or pimples. Therefore, the composition of the present disclosure has the effects of preventing hair loss, controlling oil, treating acne, and repairing.

[0169] Stability test of the Platycladus orientalis polypeptide composition of Example 19

[0170] The compositions of Examples 12 - 14 were sealed and placed under constant temperature and humidity conditions at 40°C for 1 month for accelerated stability testing. The results are shown in Table 4.

[0171] Table 4 Results of Accelerated Stability Testing

[0172]

[0173] The results showed that the combination of Platycladus orientalis leaf extract A and polypeptide had the best stability, and no solid was precipitated in the solution after 1 month of placement in the accelerated test; the stabilities of the combinations of Platycladus orientalis leaf extract B and Platycladus orientalis leaf extract C with polypeptide were relatively poor, and solids were precipitated in the solutions after 1 month of placement in the accelerated test. The results proved that there was stronger compounding stability between Platycladus orientalis leaf extract A and polypeptide, which could meet the requirements for long - term storage and use of the product.

[0174] In the present disclosure, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "comprising", "including" or any other variation thereof are intended to cover non - exclusive inclusion, such that a process, method, article or terminal device comprising a series of elements not only includes those elements but also includes other elements not expressly listed, or further includes elements inherent to such process, method, article or terminal device. Without further limitation, an element defined by the phrase "comprising an..." does not exclude the presence of additional identical elements in the process, method, article or terminal device comprising the element.

[0175] Although specific embodiments of the present disclosure have been described for purposes of illustrative explanation, those skilled in the art can make various modifications or improvements without departing from the spirit and scope of the present disclosure. These modifications or improvements shall fall within the scope of the appended claims of the present disclosure.

Claims

1. A composition, characterized in that, It contains Platycladus orientalis leaf extract and polypeptides, and the polypeptides contain Tripeptide-1, Palmitoyl Tripeptide-1 and Acetyl Tetrapeptide-3.

2. The composition according to claim 1, wherein The Platycladus orientalis leaf extract is prepared by successively extracting Platycladus orientalis leaves with a 70% ethanol solution, adsorbing with macroporous resin, and performing water precipitation.

3. The composition according to claim 2, wherein The preparation method of the Platycladus orientalis leaf extract includes the following steps: S1. Reflux extract Platycladus orientalis leaves with a 70% ethanol solution, filter, and take the extract; S2. Concentrate the extract to remove alcohol and then filter it. Load it onto AB-8 macroporous resin, first wash the impurities with water, and then elute with a 70% ethanol solution, and collect the eluate; S3. Concentrate the eluate to remove alcohol, dissolve it in water, let it stand, filter, and take the filtrate; S4. Add 1,3-propanediol and PEG-40 hydrogenated castor oil to the filtrate to obtain the Platycladus orientalis leaf extract; the volume ratio of the filtrate, 1,3-propanediol to PEG-40 hydrogenated castor oil is 19:20:

1.

4. Use of the composition according to any one of claims 1-3 in the preparation of a cosmetic composition for supplementing scalp nutrition, promoting hair growth or preventing hair loss.

5. Use of the composition according to any one of claims 1-3 in the preparation of a cosmetic composition for promoting collagen synthesis, increasing skin elasticity or improving skin firmness.

6. Use of the composition according to any one of claims 1-3 in the preparation of a cosmetic composition for controlling oil, treating acne or repairing.

7. Use of the composition according to any one of claims 1-3 in the preparation of a cosmetic composition for inhibiting the expression of acetyl-CoA carboxylase.

8. Use of the composition according to any one of claims 1-3 in the preparation of a cosmetic composition for inhibiting skin oil synthesis or secretion.

9. Use of the composition according to any one of claims 1-3 in the preparation of a cosmetic or daily chemical product for preventing hair loss, promoting hair growth or controlling oil.

10. The use according to claim 9, characterized in that, The cosmetic or daily chemical product includes shampoo, hair conditioner, hair care liquid, hair care essential oil, hair wash and care essence, hair oil, hair care lotion, scalp and hair nourishing product, hair essence, scalp paste, hair gel, hairspray, hair mask or eyebrow care liquid.

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