Peony extract with scalp anti-aging and anti-alopecia effects and application thereof

Abstract

The invention relates to the technical field of daily cosmetics, in particular to a peony extract with scalp anti-aging and anti-hair-loss functions and application. The peony extract disclosed by the invention is a peony flower extract, the mass fraction of peony flower peptide in the peony flower extract is greater than or equal to 10%, and the peony extract disclosed by the invention is found to have the effects of improving the activity of superoxide dismutase (SOD) on human hair papilla cells for the first time; the positive cell rate of beta-galactosidase (SA-beta-Gal) of human hair papilla cells is inhibited, and the functions of senescent cells are reduced; meanwhile, the functions of promoting hair follicles to express growth factors and promoting hair growth are achieved. The method can be applied to preparation of scalp care products, and the anti-aging or anti-hair-loss function of scalp is achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of daily cosmetics, and in particular to a peony extract having the effects of anti-scalp aging and anti-hair loss and an application thereof. Background Art

[0002] Peony (Paeonia suffruticosa), a plant of the genus Paeonia in the Ranunculaceae family, has a long history of being used as an ornamental and medicinal plant.

[0003] The catalog of ingredients currently used in cosmetics lists eight ingredients related to peonies: peony root bark extract, peony flower water, peony extract, peony seed oil, peony root extract, peony root water, peony flower / leaf / root extract, and peony branch / flower / leaf extract. The first three are widely used in cosmetics. Peony-related products are not only diverse in form but also offer a wide range of benefits, including skincare, whitening, anti-inflammatory, and antioxidant properties, demonstrating promising prospects. However, the efficacy and applications of peony flower extract are limited.

[0004] With the accelerating pace of modern life, scalp aging and hair loss are becoming increasingly serious. Factors such as stress, environmental pollution, and unhealthy lifestyles accelerate scalp aging, leading to hair follicle atrophy and, in turn, hair loss. According to statistics, the number of people suffering from hair loss is increasing year by year worldwide, and scalp health issues have become a major concern for many people.

[0005] In this context, it is particularly necessary to develop natural plant raw materials for scalp care. Peony flowers are rich in flavonoids, phenols and other active ingredients. These ingredients have multiple functions such as antioxidant and anti-inflammatory effects, laying a solid foundation for their application in the field of daily chemicals. At present, peony flowers have shown certain application potential in anti-aging skin care products, perfumes and other daily chemical products. For example, patent 202311551958.5 discloses "A kind of Danfeng peony petal short peptide protein and its application". This invention uses Danfeng peony petals as raw materials, obtains plant protein by alkali dissolution and acid precipitation, and hydrolyzes it with food-derived hydrolases to obtain low-molecular-weight protein polypeptides with good α-glucosidase activity, and can improve the hyperglycemia symptoms of type 2 diabetic mice, and has certain antioxidant effects. However, the application of peony flower extracts in scalp care products is still in its infancy. Summary of the Invention

[0006] Addressing a gap in the prior art, the present invention discloses a peony extract, specifically a peony flower extract. This peony extract is first shown to increase superoxide dismutase (SOD) activity in human dermal papilla cells, inhibit the rate of β-galactosidase (SA-β-Gal)-positive cells, and reduce senescent cells. It also promotes the expression of growth factors in hair follicles, thereby promoting hair growth. This extract can be used in the preparation of scalp care products to achieve anti-aging or anti-hair loss benefits. The present invention specifically addresses the following: In a first aspect, the present invention provides a peony extract, which is a peony flower extract, wherein the mass fraction of peony flower peptides in the peony flower extract is ≥10%, optionally, the mass fraction of peony flower peptides is ≥15%.

[0007] Furthermore, the peony flower peptide is a small molecule polypeptide. Optionally, the molecular weight of the peony flower peptide is ≤5000Da. Optionally, the molecular weight of the peony flower peptide is ≤2500Da. Optionally, the molecular weight of the peony flower peptide is ≤1000Da.

[0008] The second aspect of the present invention provides a composition comprising the peony extract and an additive acceptable in the cosmetics field. Optionally, the additive is at least one of a surfactant, an emulsifier, silicone oil, a moisturizer, a preservative, and a fragrance.

[0009] Furthermore, the additives include at least one of ammonium polyacryloyldimethyltaurate, dimethylsiloxane, PEG-40 hydrogenated castor oil, carbomer, cocamidopropyl betaine, and glucosidol.

[0010] Furthermore, the mass fraction of the peony extract in the composition is 0.0001% to 20%, preferably, the mass fraction of the peony extract is 0.001 to 5%, and more preferably, the mass fraction of the peony extract is 0.001 to 2%.

[0011] The third aspect of the present invention provides the use of the peony extract or the composition in any of the following aspects: A1. Application in the preparation of products for inhibiting the β-galactosidase (SA-β-Gal) positive cell rate in human dermal papilla cells; A2. Application in the preparation of products for delaying and / or inhibiting the aging of human dermal papilla cells; A3. Application in the preparation of products for increasing superoxide dismutase (SOD) activity in human dermal papilla cells; A4. Application in the preparation of products that promote the expression of growth factors in hair follicles.

[0012] The fourth aspect of the present invention provides the use of the peony extract or the composition in any of the following aspects: B1. Application in the preparation of anti-aging products; B2. Application in the preparation of products for inhibiting and / or preventing hair loss; B3. Application in the preparation of hair growth promoting products; B4. Application in the preparation of antioxidant products.

[0013] Furthermore, the product is a cosmetic. Optionally, the product includes at least one of a scalp care product and a skin care product. Optionally, the product includes at least one of a shampoo, a conditioner, a hair mask, a scalp pre-wash or a scalp essence.

[0014] In a fifth aspect, the present invention provides a method for preparing the peony extract, the method comprising at least the following steps: S1: Crush the peony flowers, add deionized water, and steam at high temperature; S2: adding protease to the peony flower aqueous solution obtained in step S1 for enzymatic hydrolysis and then filtering to obtain a filtrate containing a peony flower extract; S3: spray-drying the filtrate obtained in step S2 to obtain the peony extract.

[0015] Furthermore, the specific steps in step S1 are: After the peony flowers are crushed, deionized water is added at a material-liquid ratio of 1:10 to 1:100, and the mixture is steamed at high temperature for 20 to 180 minutes.

[0016] Furthermore, the specific steps in step S2 are: The added protease is one or more of alkaline protease, neutral protease and acidic protease, and the filtration condition is 200-5000Da.

[0017] In one embodiment of the present invention, the method for preparing the peony extract comprises the following steps: crushing peony flowers, adding deionized water at a material-to-liquid ratio of 1:30, and steaming at high temperature for 120 minutes. After adding neutral protease for a period of time, the mixture is filtered through a 1000 Da membrane and spray-dried to obtain an example sample.

[0018] The beneficial effects of the present invention include but are not limited to: The peony extract disclosed in this paper has been shown for the first time to increase superoxide dismutase (SOD) activity in human dermal papilla cells, inhibit the rate of β-galactosidase (SA-β-Gal)-positive cells, and reduce senescent cells. It also promotes the expression of growth factors in hair follicles, thereby promoting hair growth. It can be used in scalp care products to achieve anti-aging or hair loss benefits. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The drawings described herein are used to provide a further understanding of the present invention and constitute a part of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the drawings: Figure 1 Schematic diagram of the inhibitory effect of peony extract on superoxide dismutase (SOD) in human dermal papilla cells in an embodiment of the present invention.

[0020] Figure 2 Schematic diagram of the results of human dermal papilla cells expressing β-galactosidase in an embodiment of the present invention.

[0021] Figure 3 Schematic diagram of VEGF expression in hair follicles in Examples of the present invention and Comparative Examples.

[0022] Figure 4 Schematic diagram of the results of vibrissae follicle length in the examples of the present invention and the comparative examples. DETAILED DESCRIPTION

[0023] The present invention is described in detail below with reference to the examples, but the present invention is not limited to these examples. Unless otherwise specified, the raw materials and catalysts in the examples of the present invention are purchased through commercial channels.

[0024] Example 1 Preparation of Peony Extract Example Sample: Peony flowers were crushed and added to deionized water at a material-to-liquid ratio of 1:30. The mixture was then steamed at high temperature for 120 minutes. Neutral protease was added for a period of time, followed by centrifugation (4000-8000 rpm, 10-15 minutes), and the pH was adjusted to neutral. The mixture was then filtered through a 1000 Da membrane and spray-dried to obtain the Example Sample.

[0025] Comparative example sample: peony flower powder and purified water in a ratio of 1:20, heated in a water bath at 90℃ for 20 minutes, centrifuged at 3600rpm for 20 minutes, concentrated by vacuum rotary evaporation at 80℃, frozen at -18℃ overnight, and vacuum dried for 48 hours.

[0026] Peptide content comparison: Test method: Agilent 1260 Infinity II, mobile phase: 0.1 M NaNO3, detector: RID, chromatographic column: Waters Ultrahydrogel 1000 and Waters Ultrahydrogel 120 in series, temperature: 40°C, flow rate: 0.8 ml / min, standard: polyethylene oxide / polyethylene glycol PEO / PEG, injection volume: 100 μl.

[0027] Peptide content of the example sample: 23.6%.

[0028] Peptide content of comparative sample: 6.2%.

[0029] Example 2 DPPH Antioxidant Test 1. Experimental Methods 100 μL of test substance and 100 μL of DPPH were added to a 96-well plate in sequence, with 3 replicate wells set up for each reaction, and the reaction was shaken at room temperature for 5 min.

[0030] At the same time, a blank control and a self-control group were set up; 100 μL DPPPH working solution + 100 μL pure water solution was the blank control group; 100 μL sample solution + 100 μL pure water solution was the self-control group; and the absorbance value was measured at a wavelength of 517 nm using an enzyme reader.

[0031] The clearance rate was calculated according to the following formula:

[0032] Where: A t -The absorbance value of the sample tube, that is, the absorbance value of the solution after the sample reacts with DPPH A r - Absorbance value of self-control group A0-average absorbance value of blank control group Experimental Results The results are shown in Table 1. As can be seen from Table 1, the antioxidant effect of the peony extract of the present invention is significantly better than that of the conventional water-extracted peony extract.

[0033] Table 1 DPPH clearance results

[0034] Example 3 Inhibitory effect of peony extract on superoxide dismutase (SOD) in human dermal papilla cells 1. Experimental Methods This test is based on a DHT-stimulated human dermal papilla cell model. After sample administration, the antioxidant efficacy of the sample is evaluated by detecting the superoxide dismutase (SOD) content. The specific experimental process is as follows: 1) Cell seeding: 1×10 6 Cells were seeded into 6-well plates at a seeding density of 100 cells / well and cultured overnight in a DMEM / F12 medium + 10% fetal bovine serum (FBS) in an incubator (37°C, 5% CO2).

[0035] 2) Solution preparation: Prepare sample working solutions of different concentrations according to the test concentration setting table (Table 2).

[0036] 3) Drug administration: Add drugs according to Table 2 and continue incubation at 37°C, 5% CO2 for 24 h.

[0037] 4) Sample collection: Add 0.1 mL of RIPA lysis buffer (containing protease / phosphatase inhibitors) to each well and lyse the cells by pipetting. Collect the cell fluid in an EP tube and use it as the sample for testing. Store in a -80℃ freezer.

[0038] 5) Detection: Superoxide dismutase activity was detected and analyzed according to the instructions of the superoxide dismutase detection kit (purchased from Shanghai Jinpan Biotechnology Co., Ltd.).

[0039] Table 2 Concentration setting table

[0040] 2. Experimental Results The results are as follows Figure 1 As shown, the significance of the comparison between the NC group and the BC group is indicated by #, p-value < 0.05 is indicated by #, and p-value < 0.01 is indicated by ##; the significance of the comparison between the sample group and the PC group and the NC group is indicated by *, p-value < 0.05 is indicated by *, and p-value < 0.01 is indicated by **. Compared with the control group, SOD activity in human dermal papilla cells was significantly reduced after DHT stimulation, but it was significantly increased after treatment with the peony extract, indicating that the peony extract in Example 1 has antioxidant capacity, reduces oxidative stress, and delays scalp cell aging.

[0041] Example 4: Test on the Anti-hair Loss Effect of Peony Extract 1. Experimental Methods Studies have shown that the aging of hair follicle stem cells causes the hair follicles themselves to gradually atrophy, leading to hair thinning and loss. Therefore, inhibiting the aging of hair follicle stem cells can delay hair loss. β-galactosidase (SA-β-Gal) is a commonly used marker of cellular senescence, showing increased activity during cellular senescence. By measuring the activity level of SA-β-Gal in cells, the degree of cellular senescence can be assessed.

[0042] This test is based on a DHT-stimulated human dermal papilla cell model. SA-β-Gal staining is used to detect changes in SA-β-Gal expression in human dermal papilla cells to evaluate whether the test sample has an anti-hair loss effect. The specific experimental process is as follows: 1) Cell seeding: At an appropriate seeding density (8×10 4 Human dermal papilla cells were seeded into 24-well plates (100 μl / well) and incubated overnight in a culture incubator (37°C, 5% CO2) with DMEM / F12 medium + 10% fetal bovine serum (FBS).

[0043] 2) Administration: Add 1 mL of culture medium containing samples at different concentrations according to Table 2.

[0044] 3) Staining: Staining was performed according to the instructions of the β-galactosidase staining kit (Beyotime SA-β-Gal Staining Kit (C0602)).

[0045] 4) Result analysis: SA-β-Gal positive cells were quantitatively analyzed using Image Pro Plus software.

[0046] 2. Experimental Results The results are as follows Figure 2 The results are shown in the table below. Significance is indicated by * when comparing the sample group with the BC group, with p-values ​​< 0.05 indicated by *, and p-values ​​< 0.01 indicated by **. Based on a DHT-stimulated human dermal papilla cell model, the sample significantly inhibited the SA-β-Gal-positive cell rate in human dermal papilla cells at three concentrations, with statistically significant differences compared to the NC group (p < 0.05), demonstrating its anti-hair loss effect.

[0047] Example 4 Peony extract promotes hair follicles to express growth factors and promotes hair growth 1. Experimental animals and environment 1) In vitro hair follicle experiment: Animal species and strain: SPF-grade C57BL / 6J mice (age: 5 weeks, sex: male); Number of animals: 10; Quality Certificate Number: 440072000142022.

[0048] 2) Animal Source: Guangdong Medical Laboratory Animal Center: Production License No.: SCXK (Yue) 2022-0002.

[0049] 3) Breeding environment: Temperature: 20-26℃, relative humidity: 30-70%.

[0050] 2. Experimental Methods In vitro hair follicle culture and hair growth experiment: The vibrissa hair growth cycle of mice is synchronized with their age. Vibrissa hair follicles isolated in vitro can be maintained in culture for up to 23 days. The use of organ culture methods to evaluate hair follicle growth is considered to have a high correlation with the in vivo system because the degree of hair growth can be observed as the sum of the functions of each cell. The in vitro model was used to study the effect of the formula on promoting hair growth. The specific experimental process is as follows: 1) Isolation of hair follicles Culture plate pretreatment: 1% agarose (300 μL / well) was pre-coated on a 24-well plate to prevent hair follicles from adhering to the wall, and the hair follicles were placed in the center (with the hair bulb facing down).

[0051] Hair follicle culture: Incubate at 37°C, 5% CO2 for 14 days with 1 mL of culture medium (Williams E medium + 2 mM L-glutamine + 10 μg / mL insulin + 10% FBS + 1% antibiotics) per well in a 24-well plate.

[0052] Sample addition: Sample addition was performed on the second day of culture. Culture media containing different concentrations (mass fraction 0.001%, 0.05%) of the example samples and comparative example samples were added to the corresponding wells. Three parallel wells were set for each concentration. The medium was changed every three days. The blank control group (BC) was added with culture media only, and the positive control group (PC) was added with culture media containing 0.2% minoxidil.

[0053] 2) VEGF Content Assay: After collecting mouse vibrissae hair follicles on day 15 of culture, each group of follicles was transferred to a cryovial, added with PBS, and stored in a refrigerator at 80°C until use. After thawing, the specimens were maintained at 2-8°C. PBS was added, and the follicles were manually ground and disrupted by repeated freeze-thaw cycles. The cells were then centrifuged (12,000 rpm, 4°C, 15 min). The supernatant was carefully collected and assayed for VEGF content using ELISA.

[0054] 3) Image analysis: The length of the antennae follicles was photographed every 7 days, and the morphological observation and measurement of the antennae follicle length were performed.

[0055] 2. Experimental Results The results are as follows Figure 3 、 Figure 4 As shown, the peony extract in the Example significantly increased VEGF content in hair follicles at concentrations of 0.001% and 0.005%, and the relative hair growth rate was significantly higher than that of the blank control, demonstrating its excellent anti-hair loss effect. Compared with the comparative example, the Example exhibited significant advantages in promoting VEGF expression and promoting hair length growth.

[0056] The foregoing is merely an embodiment of the present invention. The scope of protection of the present invention is not limited by these specific embodiments but is determined by the claims of the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, improvements, etc. made within the technical concepts and principles of the present invention are intended to be included within the scope of protection of the present invention.

Claims

1. A peony extract, characterized in that The peony extract is a peony flower extract, wherein the mass fraction of peony flower peptides in the peony flower extract is ≥10%.

2. The peony extract according to claim 1, characterized in that The peony flower peptide is a small molecule polypeptide, and the molecular weight of the peony flower peptide is ≤5000Da.

3. A composition, characterized in that The composition comprises the peony extract according to any one of claims 1 to 2 and additives acceptable in the field of cosmetics.

4. The composition according to claim 3, characterized in that The mass fraction of the peony extract in the composition is 0.0001% to 20%.

5. Use of the peony extract according to any one of claims 1-2 or the composition according to any one of claims 3-4 in any of the following aspects: A1. Application in the preparation of products for inhibiting the β-galactosidase (SA-β-Gal) positive cell rate in human dermal papilla cells; A2. Application in the preparation of products for delaying and / or inhibiting the aging of human dermal papilla cells; A3. Application in the preparation of products for increasing superoxide dismutase (SOD) activity in human dermal papilla cells; A4. Application in the preparation of products that promote the expression of growth factors in hair follicles.

6. Use of the peony extract according to any one of claims 1-2 or the composition according to any one of claims 3-4 in any of the following aspects: B1. Application in the preparation of anti-aging products; B2. Application in the preparation of products for inhibiting and / or preventing hair loss; B3. Application in the preparation of hair growth promoting products; B4. Application in the preparation of antioxidant products.

7. The use according to any one of claims 5-6, characterized in that: The product is a cosmetic.

8. A method for preparing the peony extract according to any one of claims 1-2, characterized in that: The method comprises at least the following steps: S1: Crush the peony flowers, add deionized water, and steam at high temperature; S2: adding protease to the peony flower aqueous solution obtained in step S1 for enzymatic hydrolysis and then filtering to obtain a filtrate containing a peony flower extract; S3: spray-drying the filtrate obtained in step S2 to obtain the peony extract.

9. The method according to claim 8, characterized in that The specific steps in step S1 are: After the peony flowers are crushed, deionized water is added at a material-liquid ratio of 1:10 to 1:100, and the mixture is steamed at high temperature for 20 to 180 minutes.

10. The method according to claim 8, characterized in that The specific steps in step S2 are: The added protease is one or more of alkaline protease, neutral protease and acidic protease, and the filtration condition is 200-5000Da.

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