Application of bakuchiol in resisting androgen alopecia

Abstract

The invention discloses an application of bakuchiol in resisting androgen alopecia. According to the research result of the invention, the bakuchiol is proved to be capable of improving the expression of VEGF (vascular endothelial cell factor) of hair papilla cells, improving the signal expression of beta-catenin (catenin), inhibiting the expression of AR (androgen receptor), inhibiting the expression of TGF-beta2 and inhibiting the expression of NOX4 (nicotinamide adenine dinucleotide phosphate oxidase); the traditional Chinese medicine composition has a good anti-alopecia effect.

Description

Technical Field

[0001] The present invention relates to the use of bakuchiol, and particularly to the use of bakuchiol in anti-androgenic alopecia. Background Art

[0002] Hair is a keratin fiber formed by the self-assembly of keratin differentiated from hair follicle tissue and is an appendage of the skin. Hair plays a certain role in protecting the head from ultraviolet damage, absorbing the oil and moisture of the skin, and buffering external force damage. Hair is also an important part of human aesthetics.

[0003] Generally, hair grows vigorously in youth, with a thick diameter and a smooth and shiny appearance; as people age, the hair growth rate will gradually decline, the diameter of the hair will gradually become thinner in some people, and some people will experience hair loss. In the fast-paced working environment of modern society, the number of people with hair loss is increasing, and it is getting younger.

[0004] According to current research results, except for hair loss caused by a few immune diseases, the causes of hair loss usually include: (1) Androgenic alopecia, also known as seborrheic alopecia, the main pathogenesis is: androgens are converted into dihydrotestosterone (hereinafter referred to as DHT) by 5α-reductase. After DHT binds to the androgen receptor of dermal papilla cells, it disrupts the signal transcription of protein synthesis, reduces the release of vascular endothelial growth factor, and gradually causes hair follicle atrophy. DHT also promotes the activity of sebaceous gland cells, releases a large amount of oil into the hair follicle, squeezes the hair growth space, and also causes the overgrowth of Malassezia to release inflammatory factors, further causing hair loss. Androgenic alopecia mostly occurs in middle-aged to elderly men. The androgen receptor and 5α-reductase on the top of men's heads are more abundant than those of women. About 70% of the hair loss population are men. Women also have strong sebum secretion on the head and sparse hair on the top of the head, but women generally do not have the phenomenon of large-area baldness like men.

[0005] (2) Telogen effluvium. Growing hair suddenly enters the telogen phase, and a large number of hairs in the telogen phase fall out. Telogen effluvium is usually caused by various stress factors, such as postpartum hair loss, thyroid diseases, drug or surgical treatment, viral infections, etc.

[0006] (3) Anagen effluvium. Growing hair suddenly enters the telogen phase, and a large number of hairs in the telogen phase fall out. Telogen effluvium is usually caused by various stress factors, such as postpartum hair loss, thyroid diseases, drug or surgical treatment, viral infections, etc.

[0007] According to market statistics, more than 70% of hair loss cases are androgenetic alopecia, mainly caused by dihydrotestosterone (DHT). Therefore, researching and finding ingredients that can counteract the effects of DHT on hair follicles and hair growth, especially natural ingredients, is an important direction for intervening in and preventing androgenetic alopecia.

[0008] Currently, the common ingredients mainly include the following two synthetic pharmaceutical ingredients: (1)Finasteride. Finasteride belongs to androgen inhibitors and is also a prescription drug. The molecular structure conformation of finasteride is similar to that of androgens, and it competitively binds to 5α-reductase preferentially, thereby preventing androgens from being converted into DHT, achieving the effect of controlling androgenetic alopecia. However, long-term oral administration of finasteride may have some systemic side effects, including the regression of male secondary sexual characteristics, decreased libido, and a small number of cases may have suicidal tendencies, etc.

[0009] (2)Minoxidil. Minoxidil is a drug for treating hypertension and is used to treat hair loss. Its main functions include dilating the scalp blood vessels, increasing blood circulation, and providing more nutrients for hair follicles. Thus, it stimulates hair growth. As a drug, minoxidil also has some typical side effects: Since some minoxidil formulations require the use of a large amount of solvents (propylene glycol or ethanol), some patients have allergic symptoms, including itching, dandruff, erythema and other symptoms. Long-term use of minoxidil may cause hirsutism in other parts of some patients. Summary of the Invention

[0010] Object of the Invention: The object of the present invention is to overcome the disadvantage of unsatisfactory anti-androgenetic alopecia effect in the prior art and provide the use of bakuchiol in anti-androgenetic alopecia.

[0011] Technical Solution: The present invention relates to the use of bakuchiol in anti-androgenetic alopecia.

[0012] Further, the mass addition range of bakuchiol is: 0.01 - 0.50%.

[0013] Further, the bakuchiol is extracted from the plant Psoralea corylifolia L. of the Leguminosae family.

[0014] Further, the product is a scalp care product.

[0015] Further, the product is an anti-hair loss product.

[0016] Further, the product dosage form includes aqueous solution, emulsion or oil.

[0017] The second aspect of the present invention is to provide the use of a product containing an effective amount of bakuchiol in anti-androgenetic alopecia.

[0018] The present invention is to find a natural ingredient that can counteract the negative effects of dihydrotestosterone (DHT) on hair follicles. The ingredient of the present invention refers to bakuchiol extracted from the leguminous plant Psoralea corylifolia.

[0019] Bakuchiol, molecular weight: 256.38, and its molecular structure is shown as follows:

[0020] Bakuchiol, as a natural ingredient, has the effect of retinol when used in skin care according to literature reports. A study in 2011 showed that the use of 1% bakuchiol was superior to 2% salicylic acid in the treatment of acne. A study in 2019, a randomized double-blind experiment of 0.5% bakuchiol and 0.5% retinol, obtained similar effects in anti-wrinkle and pigmentation improvement, and the irritation was lower than that of retinol. Thus, bakuchiol is a relatively mild skin care active ingredient. However, there is no report on the use of bakuchiol for hair and scalp care, especially for anti-hair loss research.

[0021] Compared with the prior art, the present invention has the following beneficial effects: According to the research results of the present invention, it is proved that bakuchiol can increase the expression of VEGF (vascular endothelial growth factor) in dermal papilla cells, increase the expression of β-catenin signal, inhibit the expression of AR (androgen receptor), inhibit the expression of TGF-β2, and inhibit the expression of NOX4 (nicotinamide adenine dinucleotide phosphate oxidase). Specific embodiments

[0022] To make the objectives, technical solutions and advantages of the present invention clearer, the technical solutions of the present invention will be further described below.

[0023] 1. Experimental principle Dermal papilla cells (DPCs) are an important part of hair follicles, located at the bottom of hair follicles, and can regulate the hair follicle growth cycle by secreting growth factors and signaling molecules. Dihydrotestosterone (DHT) is an androgen metabolite. When highly expressed in the hair follicle microenvironment, abnormal androgen levels will lead to dysregulation of the expression of hair growth-related secretory factors (such as AR, VEGF, TGF-β2, β-Catenin, NOX4, etc.) in DPCs cells, and then damage the proliferation and differentiation of hair follicle cells, resulting in a decrease in the proliferation ability of dermal papilla cells and hair follicle degeneration. After adding the test sample, by detecting the cell proliferation ability and the expression changes of related secretory factors, it is evaluated whether it can improve the hair follicle microenvironment and achieve the anti-hair loss effect by inhibiting the DHT-related signaling pathway or activating the regeneration pathway such as Wnt / β-catenin.

[0024] 2. Experimental materials 2.1 Cell line: Human primary dermal papilla cells, sourced from Shang'en Biotech.

[0025] 2.2 Complete culture medium: DMEM medium containing 10% fetal bovine serum (FBS) (FBS, Gibco, Lot: A2743112CP) 2.3 Test conditions: Incubator temperature 37 ± 1 °C, humidity 90 ± 5%, CO2 5 ± 1%.

[0026] 2.4 MTT solution: Prepare an MTT stock solution with a concentration of 5 mg / mL using PBS, filter and sterilize it with a 0.22 μm needle filter, and store it at -20 °C (MTT stock solution, Lot: 20241206).

[0027] 3. Test procedures 3.1 Cell viability test 3.1.1 Conventional cell culture. Prepare a cell suspension, inoculate the cell suspension into a 96-well cell culture plate, and culture for 18 - 24 h.

[0028] 3.1.2 Discard the original culture medium in the wells, add test samples with different concentrations to each well, and return to the incubator for incubation for 48 ± 1 h.

[0029] 3.1.3 Take out the culture plate, remove the original culture medium, add 100 μL of culture medium to each well, then add 20 μL of MTT solution, and incubate in the incubator for 3 - 4 h. Remove the liquid in the wells, add 100 μL of DMSO to each well, place it on an oscillator and shake for 10 - 15 min, and then measure the absorbance at a wavelength of 570 nm using an ELISA reader.

[0030] 3.1.4 Based on the cell viability test, select the concentration of the test sample when the cell viability is greater than 90% as: 6.6*10 -7 (M / M). This concentration is used for subsequent gene expression tests.

[0031] 3.2 Gene expression test 3.2.1 Conventional cell culture. Inoculate the cells into a 6-well cell culture plate and culture for 18 - 24 hours.

[0032] 3.2.2 Take out the culture plate, discard the original culture medium in the wells, and divide it into a negative control group (abbreviated as NC), a model control group (containing 10 μmol DHT, abbreviated as M), and a test sample group (containing 10 μmol DHT and 6.6*10 -7The (M / M) test samples (abbreviated as TA), and the positive control group (containing 10 μmol DHT and 50 μmol minoxidil, abbreviated as PC), with 3 parallels in each group; incubated for 48 h under the conditions of 5% CO2 and 37 °C.

[0033] 3.2.3 Harvest the cells, transfer them into a 1.5 mL centrifuge tube, add 1 mL of Trizol, mix well, and let stand at room temperature for 5 min. Add 0.2 mL of chloroform, shake for 15 sec, and let stand for 3 min. Centrifuge at 4 °C, 12000 rpm for 10 minutes, and take the supernatant. Add 0.5 times the volume of absolute ethanol, gently mix the liquid in the tube, and transfer it to the adsorption column CR3. Centrifuge at 4 °C, 12000 rpm for 30 sec, and discard the waste liquid. Add 500 μL of protein removal solution, centrifuge at 4 °C, 12000 rpm for 30 sec. Add 500 μL of washing solution, let stand for 2 min, centrifuge at 4 °C, 12000 rpm for 30 sec, and discard the waste liquid. Air dry, add 30 - 100 μL of RNase-Free dH2O to dissolve. Let stand at room temperature for 2 min, centrifuge at 4 °C, 12000 rpm for 2 min.

[0034] 3.3.4 Gene expression test.

[0035] 4 Test results 4.1 Results of androgen receptor (AR) expression Table 4-1 Relative expression levels of AR Experimental group Relative expression level of AR Negative control Model control DHT Sample group <![CDATA[DHT + 6.6*10 -7 (V / V)]]> # indicates a statistically significant difference compared with the negative control group (NC) (p < 0.05); * indicates a statistically significant difference compared with the model control group (M) (p < 0.05).

[0036] The experimental results show that in terms of the relative expression level of AR, DHT in the model control group can induce the expression of androgen receptor AR, helping more DHT to enter the cells, which is not conducive to the functional expression of dermal papilla and hair follicle cells. The test samples supplemented with bakuchiol have a significant decrease compared with the model control group.

[0037] 4.2 Test results of β-catenin content Table 4.2 Test results of β-catenin content Experimental group Negative control Model control DHT Sample group <![CDATA[DHT + 6.6*10 -7 (V / V)]]> # indicates a statistically significant difference compared with the negative control group (NC) (p < 0.05); * indicates a statistically significant difference compared with the model control group (M) (p < 0.05).

[0038] The experimental results show that: in terms of the test results of β-catenin content, DHT in the model control group inhibits the expression of β-catenin. β-catenin is a crucial signaling pathway for hair keratin synthesis. The test samples with bakuchiol showed a significant increase compared to the model control group.

[0039] 4.3 Test Results of TGF-β2 Content Table 4.3 Test Results of TGF-β2 Content Experimental group Negative control Model control DHT Sample group <![CDATA[DHT + 6.6*10 -7 (V / V)]]> # indicates a statistically significant difference compared with the negative control group (NC) (p < 0.05); * indicates a statistically significant difference compared with the model control group (M) (p < 0.05).

[0040] The experimental results show that: in terms of the test results of TGF-β2 content, DHT in the model control group induces an increase in TGF-β2, and TGF-β2 is considered to be one of the factors promoting follicular miniaturization. The test samples with bakuchiol showed a significant decrease in TGF-β2 expression compared to the model control group, which is beneficial to follicular health.

[0041] 4.4 Test Results of VEGF Content Table 4.4 Test Results of VEGF Content Experimental group Negative control Model control DHT Sample group <![CDATA[DHT + 6.6*10 -7 (V / V)]]> # indicates a statistically significant difference compared with the negative control group (NC) (p < 0.05); * indicates a statistically significant difference compared with the model control group (M) (p < 0.05).

[0042] The experimental results show that: in terms of the test results of VEGF content, DHT in the model control group inhibits the expression of VEGF factor, and the test samples with bakuchiol showed a significant increase compared to the model control group. This is beneficial to capillary angiogenesis and is conducive to delivering nutrients to follicular cells.

[0043] 4.5 Test Results of NOX4 Content Table 4.5 Test Results of NOX4 Content Experimental group Negative control Model control DHT Sample group <![CDATA[DHT + 6.6*10 -7 (V / V)]]> # indicates a statistically significant difference compared with the negative control group (NC) (p < 0.05); * indicates a statistically significant difference compared with the model control group (M) (p < 0.05).

[0044] The experimental results show that: in terms of the test results of NOX4 content, DHT in the model control group induces an increase in NOX4, which will generate more reactive oxygen species (ROS) and is not conducive to follicular health. The test samples with bakuchiol showed a significant decrease compared to the model control group, which is beneficial to reducing the generation of reactive oxygen species (ROS).

[0045] 5. Application Examples 5.1 A scalp essence containing 0.1% (W / W) bakuchiol was applied to the alopecia areas on the heads of volunteers. Twice a day, after 25 days of use, obvious hair growth occurred in the alopecia areas, achieving relatively good improvement.

[0046] 5.2 A scalp essence containing 0.2% (W / W) bakuchiol was applied to the alopecia areas on the heads of volunteers. Twice a day, after 45 days of use, obvious hair growth occurred in the alopecia areas, achieving relatively good improvement.

[0047] 6. Conclusions Under the conditions of this experiment, compared with the model group (M) and the negative control group (NC), the relative contents of β-catenin, IGF-1, and VEGF were significantly decreased (p < 0.05), while the relative contents of AR, TGF-β2, and NOX4 were significantly increased (p < 0.05), indicating that DHT has an interfering effect on dermal papilla cells. When the sample group was compared with the model group, the relative contents of β-catenin, IGF-1, and VEGF were significantly increased (p < 0.05), while the relative contents of AR, TGF-β2, and NOX4 were significantly decreased (p < 0.05), indicating that bakuchiol can improve the negative effects of DHT on hair follicle dermal papilla cells and has an anti-androgenetic alopecia effect.

[0048] In actual usage cases, the alopecia conditions of volunteers were improved after using scalp essences added with 0.1% (M / M) and 0.2% (M / M).

[0049] The above are only the preferred embodiments of the present invention and do not impose any limitation on the present invention. Any person skilled in the art, without departing from the scope of the technical solution of the present invention, makes any form of equivalent substitution or modification and other changes to the disclosed technical solution and technical content of the present invention, which are still within the content of the technical solution of the present invention and still fall within the protection scope of the present invention.

Claims

1. A use of bakuchiol in anti-androgenic alopecia.

2. The use according to claim 1, characterized in that: The mass addition amount of the bakuchiol in the anti-hair loss formula ranges from 0.01% to 0.50%.

3. The use according to claim 1, characterized in that: The bakuchiol is extracted from the Psoralea corylifolia plant of the leguminous family.

4. Use of a product comprising an effective amount of bakuchiol in combating androgenic alopecia.

5. The use according to claim 4, characterized in that: The product is a scalp care product.

6. The use according to claim 4, characterized in that: The product is an anti-hair loss product.

7. The use according to claim 4, characterized in that: The product dosage forms include aqueous solutions, emulsions or oily solutions.

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