A hair loss prevention composition containing oenothein B and use thereof

Abstract

This invention discloses an anti-hair loss composition containing evening primrose extract B and its application, comprising schisandrol A and evening primrose extract B, which can synergistically upregulate the expression level of β-catenin, thereby playing an anti-hair loss role, and can be used in the preparation of anti-hair loss cosmetics.

Description

Technical Field

[0001] This invention relates to an anti-hair loss composition, particularly an anti-hair loss composition containing evening primrose extract B and its application. Background Technology

[0002] Hair loss is a common physiological phenomenon, with androgenetic alopecia (AGA) accounting for 90% of cases. Dihydrotestosterone (DHT) is a steroid hormone secreted by the testes and is one of the main androgens in the human body. In the testes and prostate, 5α-reductase can convert testosterone into DHT. DHT has a stronger affinity for androgen receptors than testosterone. When it binds to the receptor, it promotes a shortening and thinning of the hair growth cycle, miniaturization and even atrophy of hair follicles, manifesting as typical androgenetic alopecia.

[0003] The Wnt signaling pathway is one of the known mechanisms of androgenetic alopecia, participating in fundamental biological processes such as cell proliferation and differentiation. In skin hair follicle tissue, this signaling pathway regulates the growth cycle of hair matrix cells, prompting hair follicles to restart the anagen phase from the telogen phase and promoting new hair growth. β-catenin plays a crucial protective role in androgenetic alopecia by activating the Wnt / β-catenin signaling pathway, promoting hair follicle regeneration and inhibiting the hair loss process.

[0004] Evening primrose extract B, CAS number 104987-36-2, molecular formula C 68 H 48 O 44 The structural formula is: Evening primrose extract B is a dimeric macrocyclic ellagitannin with a wide range of pharmacological activities, including antioxidant, anti-inflammatory, antifungal, anti-HCV, and antitumor properties. It is an effective and specific inhibitor of poly(ADP-ribose) hydrolases.

[0005] However, there are no reports in the prior art regarding anti-hair loss compositions composed of evening primrose extract B and schisandrol A. Summary of the Invention

[0006] The purpose of this invention is to provide an anti-hair loss composition containing evening primrose extract B and its application. This invention provides a novel anti-hair loss composition comprising evening primrose extract B and schisandrol A, which synergistically upregulate β-catenin expression levels, thereby preventing hair loss.

[0007] The technical solution of the present invention: a hair loss prevention composition containing evening primrose extract B, comprising schisandrol A and evening primrose extract B.

[0008] In the aforementioned anti-hair loss composition, the concentration ratio of schisandrin A to evening primrose oil B is (100~250):2.5ppm.

[0009] In the aforementioned anti-hair loss composition, the concentration ratio of schisandrol A to evening primrose oil B is 250:2.5 ppm.

[0010] The present invention also provides the use of the above-described anti-hair loss composition in the preparation of anti-hair loss products.

[0011] In the aforementioned applications, the hair loss is androgenetic alopecia.

[0012] In the aforementioned applications, the anti-hair loss composition is used in the preparation of anti-hair loss products that upregulate β-catenin expression levels.

[0013] The product in question is a cosmetic or a pharmaceutical product. The methods of administration for cosmetics and pharmaceuticals may include oral administration, sublingual administration, topical application, spraying, injection, etc.

[0014] The present invention also provides a hair loss prevention cosmetic comprising the above-described hair loss prevention composition.

[0015] The present invention also provides a hair loss prevention medicine comprising the above-mentioned hair loss prevention composition.

[0016] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0017] This invention, through dermal papilla cell experiments, revealed that schisandrol A + evening primrose oil in the anti-hair loss composition at ratios of 250:2.5, 200:2.5, 100:2.5, 250:1, 200:1, and 100:1 (μg / mL) significantly upregulated the decrease in β-catenin caused by 100 μM DHT, thus exhibiting an anti-hair loss effect. Furthermore, when the ratio of schisandrol A + evening primrose oil in the anti-hair loss composition was between (100:2.5) and (250:2.5) μg / mL, with a CI value <1, it exhibited a synergistic effect, showing a significantly better upregulation effect on β-catenin than schisandrol A and evening primrose oil alone. Therefore, schisandrol A + evening primrose oil can be used in combination in the preparation of cosmetics with anti-hair loss effects. Attached Figure Description

[0018] Figure 1 This is a bar chart showing the results of dihydrotestosterone (DHT) concentration assay for dermal papilla cell viability.

[0019] Figure 2 This is a graph showing the results of dermal papilla cell viability assays at different concentrations of evening primrose extract B. In the graph, * indicates a significant difference compared to the control group, "*" indicates P-value < 0.05, and "**" indicates P-value < 0.01.

[0020] Figure 3 This is a graph showing the results of hair papilla cell viability testing for different ratios of the anti-hair loss composition.

[0021] Figure 4 These are immunofluorescence staining images of β-catenin on dermal papilla cells from different groups.

[0022] Figure 5 This is a statistical graph showing the fluorescence intensity of β-catenin in dermal papilla cells from different groups. In the graph, # indicates a significant difference compared to the blank control group, "####" indicates P-value < 0.0001; * indicates a significant difference compared to the negative control (DHT) group, "*" indicates P-value < 0.05, "**" indicates P-value < 0.01, "***" indicates P-value < 0.001, and "****" indicates P-value < 0.0001. Detailed Implementation

[0023] The present invention will be further described below with reference to the accompanying drawings and embodiments, but this should not be construed as limiting the present invention.

[0024] In the description of this invention, it should also be noted that, unless otherwise explicitly specified and limited, the terms "set," "install," "connect," and "link" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, a hinged connection, a rotatable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium, or a connection within two components. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0025] Example:

[0026] 1. Experimental Methods:

[0027] 1.1) After resuscitating dermal papilla cells (DP cells), cell growth was observed. Cells were counted when the cell coverage reached 80% or more, and then seeded into 96-well and 24-well plates. The 96-well plates were used to detect cell viability, and the 24-well plates were used to detect β-catenin expression levels. The plates were incubated overnight in a CO2 incubator (37°C, 5% CO2).

[0028] 1.2) Solution preparation: Prepare the working solution for the sample.

[0029] 1.3) When the DP cell plating rate reaches about 60%, administer the drug to groups. Each group has 3 replicates.

[0030] 1.3.1) Dihydrotestosterone (DHT) modeling experiment:

[0031] The concentrations of dihydrotestosterone (DHT) for modeling experiments were explored according to the groups in Table 1 below. The DHT concentrations ranged from 1 to 1000 μM. After administration, the 96-well plates were placed in an incubator (37°C, 5% CO2) and incubated for 24 hours. Cell viability was then detected.

[0032] Table 1. Concentration grouping of dihydrotestosterone (DHT) modeling experiment

[0033]

[0034] 1.3.2) Results of dihydrotestosterone (DHT) modeling experiment:

[0035] The results of DHT concentration assay on dermal papilla cell viability are as follows: Figure 1 As shown, within a concentration range not exceeding 100 μM, the viability of dermal papilla cells is higher than 70%, indicating low toxicity, allowing for further experiments.

[0036] 1.3.3) DP cell experiment of evening primrose extract B:

[0037] The concentrations of evening primrose extract B in DP cells were determined according to the groups in Table 2 below. Concentrations ranging from 0.1 to 50 μg / mL were selected for the experiment. After administration, the 96-well plates were placed in an incubator (37°C, 5% CO2) and incubated for 24 hours to detect cell viability.

[0038] Table 2. Grouping of DP cell experimental concentrations of evening primrose extract B

[0039]

[0040] 1.3.4) Results of the DP cell experiment of evening primrose extract B:

[0041] The results of evening primrose extract B at different concentrations on the viability of dermal papilla cells are as follows: Figure 2 As shown, within a concentration range not exceeding 2.5 μg / mL, the viability of dermal papilla cells was higher than 90%, and there was no cytotoxicity.

[0042] 1.3.5) DP cell experiment of the anti-hair loss composition:

[0043] The DP cell experimental ratio of the anti-hair loss composition was explored according to the grouping in Table 3 below. After administration, the 96-well plate was placed in an incubator (37℃, 5% CO2) and incubated for 24 hours to detect cell viability.

[0044] Table 3. Grouping of DP cell experiments with different ratios of the compositions.

[0045]

[0046] 1.3.6) Results of DP cell experiments on the anti-hair loss composition:

[0047] Results of hair loss prevention composition on hair papilla cell viability at different formulation ratios are as follows: Figure 3 As shown, in the composition, when the ratio of schisandrin A to evening primrose oil B is in the range of (100:2.5) to (250:1) μg / mL, the viability of dermal papilla cells is higher than 90% and there is no cytotoxicity.

[0048] 1.3.7) β-catenin level detection experiment:

[0049] The optimal DHT concentration for modeling was determined to be 100 μM. β-catenin was then detected using immunofluorescence assay according to the groups grouped in Table 4 below.

[0050] Table 4. Immunofluorescence detection protocol for β-catenin

[0051]

[0052] After administration, the 24-well plate was placed in an incubator (37°C, 5% CO2) and incubated for 24 hours to detect β-catenin expression levels.

[0053] Immunofluorescence staining: Fixation, blocking, addition of primary antibody, addition of secondary antibody, counterstaining with DAPI, mounting, and then photographing using a fluorescence microscope. Quantitative analysis of fluorescence intensity was performed using Image Pro Plus software.

[0054] 1.3.6) Results of β-catenin level detection experiment:

[0055] The results of β-catenin immunofluorescence staining of dermal papilla cells in different groups are as follows: Figure 4 As shown, the statistical results of β-catenin fluorescence intensity in dermal papilla cells for different groups are as follows: Figure 5 As shown.

[0056] Based on the above results, it can be seen that 100µM DHT significantly reduced the expression level of β-catenin compared with the blank control group, with an inhibition rate of 71.42%, indicating that the model was successfully established.

[0057] Compared with the DHT group, 10 μM minoxidil significantly increased the expression level of β-catenin, with an increase of 130.94%;

[0058] Evening primrose extract at concentrations of 1 μg / mL and 2.5 μg / mL significantly upregulated the decrease in β-catenin induced by 100 μM DHT (p<0.001), with increases of 88.40% and 83.97%, respectively.

[0059] Therefore, evening primrose extract B has the effect of preventing hair loss.

[0060] The composition (schisandrin A + evening primrose extract B) significantly upregulated the reduction in β-catenin caused by 100 μM DHT at ratios of 250:2.5, 200:2.5, 100:2.5, 250:1, 200:1, and 100:1 (μg / mL) (p<0.05), with growth rates of 232.38%, 204.89%, 221.76%, 91.15%, 47.71%, and 74.88%, respectively; therefore, the composition of the present invention has an anti-hair loss effect.

[0061] Combination Effect Index (CI): The combination effect index is used to determine whether the promoting effect of the combination containing evening primrose oil and schisandrol A on β-catenin is synergistic or antagonistic. The combination effect index is calculated according to the report in the literature (Xu H, et al. Exploring the Active Constituents of Andrographis paniculata in Protecting the Skin Barrier and the Synergistic Effects with Collagen XVII. Antioxidants (Basel). 2025 Jan 20;14(1):118.), and the specific calculation formula is as follows:

[0062]

[0063]

[0064] In the formula, V m V t V C These represent the values ​​for the model group, sample group, and control group, respectively.

[0065] EAB is the combined effect of components A and B; EA is the effect of component A alone; EB is the effect of component B alone.

[0066] If CI < 1, the composition is considered to exhibit a synergistic effect.

[0067] If CI=1, the composition is considered to exhibit an additive effect.

[0068] If CI > 1, the composition is considered to exhibit antagonistic effects.

[0069] The calculated results of the combined effect index (CI) of the anti-hair loss composition (schisandrin A + evening primrose extract B) on β-catenin in hair papilla cells are shown in Table 5 below.

[0070] Table 5 CI values ​​of the compositions

[0071]

[0072] As shown in Table 5, the CI values ​​of schisandrol A and evening primrose extract at ratios of 250:2.5, 200:2.5, and 100:2.5 (μg / mL) were 0.56, 0.63, and 0.63, respectively, all <1, indicating a synergistic effect. The upregulation effect on β-catenin was significantly better than that of schisandrol A and evening primrose extract alone.

[0073] Therefore, when the ratio of schisandrol A to evening primrose extract B is between (100:2.5) and (250:2.5) (μg / mL), the anti-hair loss effect is better than that of using schisandrol A and evening primrose extract B alone, and they can be used in combination in the preparation of cosmetics with anti-hair loss effects.

[0074] The parts of this invention not described in detail are prior art and therefore will not be specifically described here.

Claims

1. A hair loss prevention composition containing evening primrose extract B, characterized in that: Including schisandrol A and evening primrose extract B.

2. The anti-hair loss composition containing evening primrose extract B according to claim 1, characterized in that: The concentration ratio of schisandrin A to evening primrose oil B is (100~250):2.5ppm.

3. The anti-hair loss composition containing evening primrose extract B according to claim 2, characterized in that: The concentration ratio of schisandrin A to evening primrose oil B is 250:2.5 ppm.

4. The use of the anti-hair loss composition according to any one of claims 1-3 in the preparation of anti-hair loss products.

5. The application according to claim 4, characterized in that: The hair loss mentioned is androgenetic alopecia.

6. The application according to claim 4, characterized in that: The application of the hair loss prevention composition in the preparation of hair loss prevention products with the function of upregulating β-catenin expression levels.

7. The application according to claim 4, characterized in that: The product in question is either a cosmetic or a pharmaceutical.

8. A hair loss prevention cosmetic, characterized in that: It comprises the anti-hair loss composition according to any one of claims 1-3.

9. A hair loss prevention medicine, characterized in that: It comprises the anti-hair loss composition according to any one of claims 1-3.

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