Use of angelica acutibaccata glycosides in the preparation of hair loss prevention products

Abstract

This invention discloses the application of angeloylgomisin H and its composition in the preparation of anti-hair loss products. Angeloylgomisin H has the effect of preventing hair loss, and the anti-hair loss composition contains angeloylgomisin H and evening primrose oil B. The two can synergistically upregulate the expression levels of VEGF and β-catenin, thereby playing a synergistic role in preventing hair loss.

Description

Technical Field

[0001] This invention relates to novel applications of angeloylgomisin H, particularly the application of angeloylgomisin H and its compositions in the preparation of anti-hair loss products. 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] Vascular endothelial growth factor (VEGF) is a key angiogenesis factor that promotes angiogenesis around hair follicles, increases blood supply to hair follicles, and thus supports hair growth. Due to the cyclical changes in VEGF, it regulates the hair follicle growth cycle. At the same time, VEGF can protect hair follicle stem cells from androgen-induced apoptosis through specific signaling pathways (such as the PI3K / AKT pathway), playing an important role in maintaining normal hair follicle function and hair growth.

[0004] 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.

[0005] In addition, inflammation is closely associated with various hair loss symptoms. Interleukin-6 (IL-6) is a key immunomodulatory cytokine that is induced by DHT in the dermis of the scalp of hair loss patients and participates in androgen-driven changes in autocrine and paracrine factors. In androgenetic alopecia, IL-6 promotes hair loss by affecting the function of hair follicle stem cells; it can also induce the transition of hair follicles from the anagen to catagen phase, inhibit hair growth, and ultimately affect hair shaft elongation and hair follicle health.

[0006] Angelica aucubin H, CAS number 66056-22-2, molecular formula C 28 H 36 O8, the structural formula is:

[0007] Angelicoylgomisin H is a lignan compound extracted from Schisandra chinensis. Its core function is liver protection, and it also has the potential to improve glucose metabolism.

[0008] 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 poly(ADP-ribose) hydrolase inhibitor.

[0009] However, there are no reports in the existing technology regarding the anti-hair loss effect of angeloylgomisin H. Summary of the Invention

[0010] The purpose of this invention is to provide an application of angeloylgomisin H and its composition in the preparation of anti-hair loss products. This invention provides a novel application of angeloylgomisin H and a novel anti-hair loss composition comprising angeloylgomisin H and evening primrose oil B, which synergistically upregulate the expression levels of VEGF and β-catenin, thereby achieving a synergistic anti-hair loss effect.

[0011] The technical solution of this invention: the application of angeloylgomisin H in the preparation of anti-hair loss products.

[0012] In the aforementioned applications, the concentration of angeloylgomisin H used is 0.5~50 ppm.

[0013] A hair loss prevention composition comprising angeloylgomisin H and evening primrose extract B.

[0014] The aforementioned anti-hair loss composition is used in the preparation of anti-hair loss products.

[0015] In the aforementioned applications, the hair loss prevention product is a hair loss prevention product that has the function of upregulating VEGF, β-catenin and / or reducing IL-6 protein expression.

[0016] In the aforementioned applications, the hair loss is androgenetic alopecia.

[0017] In the aforementioned applications, when the hair loss prevention composition is used to prepare hair loss prevention products with the function of upregulating VEGF protein expression, the concentration ratio of angeloylgomisin H and evening primrose extract B is (10~50):2.5ppm.

[0018] In the aforementioned applications, when the hair loss prevention composition is used to prepare hair loss prevention products with the function of upregulating β-catenin protein expression, the concentration ratio of angeloylgomisin H and evening primrose oil B is (25~50):2.5ppm or 50:1ppm.

[0019] In the aforementioned applications, the product is a cosmetic or a pharmaceutical. The methods of administration for cosmetics and pharmaceuticals may include oral administration, sublingual administration, topical application, spraying, injection, etc.

[0020] The present invention also provides a hair loss prevention cosmetic, comprising angeloyl gomisin H or the above-mentioned hair loss prevention composition.

[0021] The present invention also provides a hair loss prevention medicine comprising angeloylgomisin H or the above-mentioned hair loss prevention composition.

[0022] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0023] In this invention, a hair papilla cell assay revealed that angeloylgomisin H can significantly upregulate the decrease in VEGF and β-catenin caused by 100μM DHT, and significantly downregulate the increase in IL-6 caused by 100μM DHT, thereby exhibiting an anti-hair loss effect.

[0024] In the anti-hair loss composition, angeloylgomicin H + evening primrose oil B at ratios of 50:2.5, 25:2.5, and 10:2.5 (μg / mL) showed significantly better upregulation of VEGF than either angeloylgomicin H or evening primrose oil alone, with a CI value <1, indicating a synergistic effect. Similarly, at ratios of 50:1, 50:2.5, and 25:2.5 (μg / mL), angeloylgomicin H + evening primrose oil B at ratios of 50:1, 50:2.5, and 25:2.5 (μg / mL) showed significantly better upregulation of β-catenin than either angeloylgomicin H or evening primrose oil alone, with a CI value <1, indicating a synergistic effect.

[0025] Therefore, angeloylgomisin H and its composition can be used in the preparation of products with anti-hair loss effects. Attached Figure Description

[0026] Figure 1 This is a bar chart showing the results of DHT concentration assay for dermal papilla cell viability.

[0027] Figure 2 This is a graph showing the results of dermal papilla cell viability assays at different concentrations of angeloylgomisin H alone.

[0028] Figure 3 This is a graph showing the results of hair papilla cell viability testing when the anti-hair loss composition is formulated in different proportions.

[0029] Figure 4This is a graph showing the results of VEGF expression in dermal papilla cells in 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.

[0030] Figure 5 This is a graph showing the results of IL-6 expression in dermal papilla cells in different groups. In the graph, # indicates a significant difference compared to the blank control group, and "#" indicates P-value < 0.05; * indicates a significant difference compared to the negative control (DHT) group, and "*" indicates P-value < 0.05, "**" indicates P-value < 0.01, and "***" indicates P-value < 0.001.

[0031] Figure 6 These are immunofluorescence staining images of β-catenin on dermal papilla cells from different groups.

[0032] Figure 7 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

[0033] 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.

[0034] Example:

[0035] 1. Experimental Methods:

[0036] 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 the expression levels of VEGF, IL-6, and β-catenin. The plates were incubated overnight in a CO2 incubator (37°C, 5% CO2).

[0037] 1.2) Solution preparation: Prepare the working solution for the sample.

[0038] 1.3) When the DP cell plating rate reaches about 60%, administer the drug to groups. Each group has 3 replicates.

[0039] 1.3.1) Dihydrotestosterone (DHT) modeling experiment:

[0040] 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.

[0041] Table 1. Concentration grouping of dihydrotestosterone (DHT) modeling experiment

[0042]

[0043] 1.3.2) Results of dihydrotestosterone (DHT) modeling experiment:

[0044] The effects of different DHT concentrations on dermal papilla cell viability detection results are as follows: Figure 1 As shown, within the concentration range of 1~100μM, the viability of dermal papilla cells is higher than 70%, and the toxicity is relatively low, allowing for further experiments.

[0045] 1.3.3) DP cell assay with angeloylgomisin H:

[0046] The concentrations of angeloylgammadextrin H in DP cells were determined according to the groupings in Table 2 below. Concentrations ranging from 0.5 to 200 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.

[0047] Table 2. Grouping of DP cell experimental concentrations of angeloylgamma-hydroxyphenidate H.

[0048]

[0049] 1.3.4) Results of DP cell experiments with angeloylgomisin H:

[0050] The results of dermal papilla cell viability assay at different concentrations of angeloylgromyn H are as follows: Figure 2 As shown, within a concentration range not exceeding 50 μg / mL, the viability of dermal papilla cells was higher than 90%, and there was no cytotoxicity.

[0051] 1.3.5) DP cell experiment of the anti-hair loss composition:

[0052] 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.

[0053] Table 3. Grouping of DP cell experiments with different ratios of the compositions.

[0054]

[0055] 1.3.6) Results of DP cell experiments on the anti-hair loss composition:

[0056] Results of hair loss prevention composition on dermal papilla cell viability at different formulation ratios are as follows: Figure 3 As shown, in the anti-hair loss composition, when the ratio of angeloylgomisin H to evening primrose oil B is within the range of 50:1 μg / mL, the dermal papilla cell viability is higher than 90% and there is no cytotoxicity.

[0057] 1.3.7) Experimental detection of VEGF, IL-6 and β-catenin levels:

[0058] The optimal DHT concentration for modeling was determined to be 100 μM. The samples were grouped according to Table 4 below, and VEGF and IL-6 were quantitatively detected by ELISA, and β-catenin was detected by immunofluorescence.

[0059] Table 4. Experimental Protocol for Detection of VEGF, IL-6, and β-catenin Levels

[0060]

[0061] 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.

[0062] ELISA assay: Collect the supernatant and perform ELISA assay according to the ELISA kit instructions.

[0063] 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.

[0064] 1.3.7) Experimental results of VEGF, IL-6 and β-catenin level detection:

[0065] Results of VEGF expression in dermal papilla cells in different groups are as follows Figure 4 As shown, the expression results of IL-6 in dermal papilla cells in different groups are as follows: Figure 5As shown, the immunofluorescence staining results of β-catenin on dermal papilla cells in different groups are as follows: Figure 6 As shown, the statistical results of β-catenin fluorescence intensity in dermal papilla cells for different groups are as follows: Figure 7 As shown.

[0066] Based on the above results, 100 μM DHT significantly reduced the expression level of VEGF (77.19%) compared to the blank control group; significantly increased the expression level of IL-6 (7.51%); and significantly reduced the expression level of β-catenin (64.48%), indicating that the model was successfully established.

[0067] Compared with the DHT group, 10 μM minoxidil significantly increased VEGF expression level by 91.76%; significantly decreased IL-6 expression level by 33.16%; and significantly increased β-catenin expression level by 110.83%.

[0068] Angelica glycoside H at concentrations of 0.5, 1, 10, 25, and 50 μg / mL significantly upregulated the VEGF reduction induced by 100 μM DHT, with increases of 23.17%, 42.43%, 30.67%, 22.24%, and 116.46%, respectively.

[0069] Angelica oryzanol H at concentrations of 0.5, 1, 10, 25, and 50 μg / mL significantly downregulated the increase in IL-6 induced by 100 μM DHT, with inhibition rates of 18.34%, 31.04%, 21.66%, 15.73%, and 24.41%, respectively.

[0070] 0.5, 1, 10, 25 and 50 μg / mL of angeloylgomicin H significantly upregulated the decrease in β-catenin caused by 100 μM DHT, with growth rates of 89.46%, 100.92%, 85.83%, 102.48% and 39.03%, respectively.

[0071] Therefore, gonadotropin H has the effect of preventing hair loss.

[0072] 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:

[0073]

[0074]

[0075] In the formula, V m V t V C These represent the values ​​for the model group, sample group, and control group, respectively.

[0076] 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.

[0077] If CI < 1, the composition is considered to exhibit a synergistic effect.

[0078] If CI=1, the composition is considered to exhibit an additive effect.

[0079] If CI > 1, the composition is considered to exhibit antagonistic effects.

[0080] The calculated combined effect index (CI) of the composition (angeloylgomisin H + evening primrose extract B) on VEGF and β-catenin in dermal papilla cells is shown in Table 5 below.

[0081] Table 5 CI values ​​of the compositions

[0082]

[0083] according to Figure 4As shown in Table 5, in the anti-hair loss composition, angeloyl gomisin H and evening primrose extract B can significantly upregulate the concentration of hair loss from 100 μM at ratios of 50:2.5, 25:2.5, 10:2.5, 1:2.5, 0.5:2.5, 50:1, and 25:1 (μg / mL). The reduction in VEGF induced by DHT showed growth rates of 209.48%, 197.62%, 147.03%, 67.99%, 78.21%, 101.05%, and 45.72%, respectively, which was superior to that of angeloylgomicin H alone, demonstrating an anti-hair loss effect. Angeloylgomicin H combined with evening primrose oil at ratios of 50:2.5, 25:2.5, and 10:2.5 (μg / mL) showed significantly better upregulation of VEGF than angeloylgomicin H and evening primrose oil alone, with CI values ​​of 0.86, 0.57, and 0.81, respectively (CI < 1), indicating a synergistic effect.

[0084] according to Figure 7 As shown in Table 5, in the anti-hair loss composition, angeloyl gomisin H + evening primrose extract B can be significantly upregulated from 100 μM at ratios of 50:2.5, 25:2.5, 10:2.5, 1:2.5, 0.5:2.5, 50:1, and 25:1 (μg / mL). The reduction in β-catenin induced by DHT, with growth rates of 114.88%, 152.50%, 102.80%, 93.87%, 83.19%, 127.03%, and 115.83%, respectively, showed an anti-hair loss effect. The combination of angeloylgomicin H and evening primrose oil B at ratios of 50:1, 50:2.5, and 25:2.5 (μg / mL) significantly outperformed angeloylgomicin H and evening primrose oil B alone, with CI values ​​of 0.60, 0.79, and 0.86, respectively (CI < 1), indicating a synergistic effect.

[0085] The parts of this invention not described in detail are prior art and therefore will not be specifically described here.

Claims

1. Application of Angelica yauroside H in the preparation of anti-hair loss products.

2. The application according to claim 1, characterized in that: The concentration of angeloylgomisin H used is 0.5~50 ppm.

3. A hair loss prevention composition, characterized in that: Including angeloylgomisin H and evening primrose extract B.

4. The use of the anti-hair loss composition according to claim 4 in the preparation of anti-hair loss products.

5. In the application according to claim 1 or 4, the characteristic is: The hair loss prevention product is one that upregulates VEGF, β-catenin, and / or reduces the expression of IL-6 protein.

6. In the application according to claim 5, the characteristic is: In the preparation of hair loss prevention products with the function of upregulating VEGF protein expression, the concentration ratio of angeloylgomisin H and evening primrose extract B in the hair loss prevention composition is (10~50):2.5ppm.

7. In the application according to claim 5, the characteristic is: In the preparation of hair loss prevention products with the function of upregulating β-catenin protein expression, the concentration ratio of angeloylgomisin H and evening primrose oil B in the hair loss prevention composition is (25~50):2.5ppm or 50:1ppm.

8. The application according to claim 1 or 4, characterized in that: The product in question is either a cosmetic or a pharmaceutical.

9. A hair loss prevention cosmetic, characterized in that: Contains angeloylgomisin H or the anti-hair loss composition of claim 3.

10. A hair loss prevention medicine, characterized in that: Contains angeloylgomisin H or the anti-hair loss composition of claim 3.

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