A composition containing myricetin and quercetin, a cypress leaf extract containing the composition, and application thereof in promoting hair growth and preventing hair loss

Abstract

The application provides a composition containing myricetin and quercetin, a cypress leaf extract containing the composition and application of the cypress leaf extract in promoting hair growth and preventing hair loss, relates to the technical field of daily chemicals, is composed of myricetin and quercetin, and adopts liquid quality and network pharmacology methods to analyze the main acting components and proportions of the cypress leaf extract, finds that myricetin and quercetin are the main components for preventing hair loss and promoting hair growth, and explores the hair loss prevention / hair growth promotion potential of different compositions of myricetin and quercetin through a 5alpha-reductase inhibition rate experiment, so as to provide a theoretical basis for the application of the myricetin and quercetin composition.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of daily chemicals, in particular to a composition containing myricetin and five-hydroxy flavone, a cypress leaf extract containing the composition and application of the cypress leaf extract in promoting hair growth and preventing hair loss. BACKGROUND

[0002] Hair loss is a common skin disease that affects the quality of life of patients and has an impact on the psychology of patients. The cause and mechanism of hair loss have not been fully elucidated, but it is generally believed that it may be caused by both genetic and environmental stimuli.

[0003] Androgenetic alopecia (AGA), also known as seborrheic alopecia, is a common cause of hair loss. 5α-reductase controls androgen metabolism, and 5α-reductase inhibitors are used clinically to selectively block the synthesis of dihydrotestosterone by inhibiting 5α-reductase activity, thereby reducing the level of androgens in the body to treat androgenetic alopecia, such as steroidal drugs like finasteride and dutasteride. However, they often have strong side effects.

[0004] Cypress leaves are widely used in traditional Chinese medicine for treating hair loss. It is believed to cool blood, stop bleeding, clear heat and detoxify, effectively inhibit scalp inflammation, and promote hair follicle health. In particular, for hair loss problems caused by excessive oil secretion or clogged hair follicles, cypress leaf shampoo can help improve the scalp environment and promote hair growth, thereby reducing hair loss. Cypress leaves also have the function of regulating sebum secretion. However, as a natural ingredient, cypress leaves are relatively safe to use and suitable for most people. However, the active ingredients of traditional Chinese medicine are complex and diverse, and it is difficult to control the standard and identify the target.

[0005] Therefore, the present application is proposed. SUMMARY

[0006] One of the purposes of the present application is to provide a composition to at least solve one of the technical problems in the prior art.

[0007] The second purpose of the present application is to provide a cypress leaf extract.

[0008] The third purpose of the present application is to provide a preparation method of the cypress leaf extract as described above.

[0009] The fourth purpose of the present application is to provide the application of the composition as described above, the cypress leaf extract as described above and the cypress leaf extract prepared by the preparation method as described above in the preparation of a product for promoting hair growth and / or preventing hair loss.

[0010] The fifth purpose of the present application is to provide the application of the composition as described above in the preparation of a product for inhibiting 5α-reductase.

[0011] In order to achieve the above object of the present application, the following technical solutions are adopted:

[0012] In a first aspect, the present application provides a composition consisting of myricetin and quercetin.

[0013] Further, the weight ratio of myricetin and quercetin is 1-14:1.

[0014] In a second aspect, the present application provides a cypress leaf extract containing the above-mentioned composition, wherein the composition accounts for ≥0.003% of the cypress leaf extract by weight.

[0015] Further, the weight ratio of myricetin and quercetin in the composition is 6.1-12.5:1.

[0016] In a third aspect, the present application provides a preparation method of the above-mentioned cypress leaf extract, comprising uniformly mixing cypress leaf coarse powder and an extraction solvent in a volume ratio of 1:5-17, soaking, ultrasonicating, and filtering to obtain the cypress leaf extract.

[0017] The extraction solvent comprises water or an alcohol aqueous solution.

[0018] Further, the volume ratio of the cypress leaf coarse powder and the extraction solvent is 1:15.

[0019] Further, the soaking time is at least 1 h.

[0020] Further, the ultrasonicating time is 20-50 min.

[0021] Further, the ultrasonicating and filtering are performed 1-2 times, and the filtrates are combined after filtering to obtain the cypress leaf extract.

[0022] The particle size of the cypress leaf coarse powder is 40-60 mesh.

[0023] In a fourth aspect, the present application provides the use of the above-mentioned composition, the above-mentioned cypress leaf extract, and the cypress leaf extract prepared by the above-mentioned preparation method in the preparation of a product for promoting hair growth and / or preventing hair loss.

[0024] In a fifth aspect, the present application provides the use of the above-mentioned composition in the preparation of a product for inhibiting 5α-reductase.

[0025] The composition provided by the present application uses liquid chromatography and network pharmacology to analyze the dominant components and proportions of cypress leaf extract, and finds that myricetin and quercetin are the dominant components for preventing hair loss and promoting hair growth. The 5α-reductase inhibition rate experiment is used to explore the potential of myricetin and quercetin in different compositions for preventing hair loss and promoting hair growth, thereby providing a theoretical basis for the application of myricetin and quercetin composition. BRIEF DESCRIPTION OF DRAWINGS

[0026] In order to more clearly illustrate the technical solutions in the specific embodiments or prior art of the present application, the drawings required to be used in the description of the specific embodiments or prior art will be briefly introduced as follows. Obviously, the drawings in the following description are some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without any creative effort on the basis of these drawings.

[0027] Figure 1 Effects of the Cacumen Platycladi alcohol extract and the Cacumen Platycladi water extract provided for Embodiment 1 of the present application on the viability of human hair papilla cells;

[0028] Figure 2 Effects of the Cacumen Platycladi alcohol extract provided for Embodiment 1 of the present application on hair growth in mice;

[0029] Figure 3 Negative ion total ion current chromatogram of the Cacumen Platycladi alcohol extract provided for Embodiment 2 of the present application;

[0030] Figure 4 Positive ion total ion current chromatogram of the Cacumen Platycladi alcohol extract provided for Embodiment 2 of the present application;

[0031] Figure 5 Negative ion total ion current chromatogram of the Cacumen Platycladi water extract provided for Embodiment 2 of the present application;

[0032] Figure 6 Positive ion total ion current chromatogram of the Cacumen Platycladi water extract provided for Embodiment 2 of the present application;

[0033] Figure 7 Cacumen Platycladi component-hair loss target point Venn diagram provided for Embodiment 3 of the present application;

[0034] Figure 8 Cacumen Platycladi extract active anti-hair loss / hair growth promotion core target point network diagram provided for Embodiment 3 of the present application. DETAILED DESCRIPTION

[0035] Unless otherwise defined, scientific and technical terms used in connection with the present application shall have the meanings that are commonly understood by those of ordinary skill in the art. The meaning and scope of the terms should be clear; however, in the event of any latent ambiguity, the definitions provided herein take precedence over any dictionary or extrinsic definition. In this application, unless indicated otherwise, the use of "or" means "and / or". Furthermore, the use of the term "including" as well as other forms such as "include", "includes" for indicating the presence of one or more elements is non-limiting.

[0036] The technical solutions of the present application will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are only some of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative efforts belong to the scope of the present application.

[0037] In one aspect of the present application, a composition is provided, which consists of myricetin and quercetin.

[0038] The inventors analyzed the dominant components and proportions of the Platycladus orientalis extract by liquid chromatography and network pharmacology methods, and found that myricetin and quercetin are the dominant components for preventing hair loss and promoting hair growth. The anti-hair loss / hair growth potential of different combinations of myricetin and quercetin was explored through 5a-reductase inhibition rate experiments, providing a theoretical basis for the application of myricetin and quercetin combinations.

[0039] In some specific embodiments, the weight ratio of myricetin to quercetin is 1-14:1.

[0040] The weight ratio of myricetin to quercetin can be, but is not limited to, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1 or 14:1, or any value between 1-14:1, preferably 1-10:1.

[0041] According to another aspect of the present application, a Platycladus orientalis extract is also provided, which comprises the above-mentioned composition, and the composition accounts for ≥0.003% of the weight ratio of the Platycladus orientalis extract, preferably ≥0.03%, more preferably ≥0.3%, and further preferably ≥0.5%.

[0042] In some specific embodiments, the weight ratio of myricetin to quercetin in the composition is 6.1-12.5:1.

[0043] The weight ratio of myricetin to quercetin in the composition can be, but is not limited to, 6.1:1, 6.5:1, 7:1, 7.5:1, 8:1, 8.5:1, 9:1, 9.5:1, 10:1, 10.5:1, 11:1, 11.5:1, 12:1 or 12.5:1, or any value between 6.1-12.5:1.

[0044] According to another aspect of the present application, a preparation method of the above-mentioned Platycladus orientalis extract is also provided, which comprises uniformly mixing Platycladus orientalis coarse powder and an extraction solvent in a volume ratio of 1:5-17, soaking, ultrasonic treatment and filtering to obtain the Platycladus orientalis extract.

[0045] The extraction solvent comprises water or an alcohol-water solution.

[0046] In some embodiments, the aqueous alcohol solution comprises 95% ethanol, 70% ethanol, 30% ethanol, 70% butylene glycol, 30% butylene glycol, 70% glycerol, 30% glycerol, or any of the supramolecular solutions comprising alcohol.

[0047] In some embodiments, the volume ratio of the crude Plume Poplar leaves to the extraction solvent is 1:15. In some embodiments, the soaking time is at least 1 h. In some embodiments, the ultrasonic time is 20-50 min; in some embodiments, the number of times of ultrasonic and filtration is 1-2 times, and the filtrate is combined after filtration to obtain the Plume Poplar leaf extract; in some embodiments, the particle size of the crude Plume Poplar leaves is 40-60 mesh.

[0048] According to another aspect of the present application, the above-mentioned composition, the above-mentioned Plume Poplar leaf extract, and the Plume Poplar leaf extract prepared by the above-mentioned method are also provided for use in the preparation of a product for promoting hair growth and / or preventing hair loss.

[0049] According to another aspect of the present application, the above-mentioned composition is also provided for use in the preparation of a product for inhibiting 5α-reductase.

[0050] The present application is further illustrated by the following examples. Unless otherwise specified, the materials in the examples are prepared according to the existing methods, or directly purchased from the market.

[0051] Example 1 Evaluation of the anti-hair loss and hair growth promoting effects of Plume Poplar leaf ethanol extract and water extract

[0052] 1. Preparation of Plume Poplar leaf ethanol extract and Plume Poplar leaf water extract

[0053] The dry Plume Poplar leaf powder crushed to 40-60 mesh is mixed with 95% ethanol or deionized water at a solid-liquid ratio of 1:15, stirred uniformly, soaked at room temperature for 1 h, placed in an ultrasonic instrument for 30 min, filtered, and the filtrate is centrifuged and then repeatedly extracted twice. The filtrate is combined and concentrated on a rotary evaporator, and stored at -20°C to obtain the Plume Poplar leaf ethanol extract and Plume Poplar leaf water extract.

[0054] 2. Effect of Plume Poplar leaf ethanol extract and Plume Poplar leaf water extract on the proliferation of human hair papilla cells

[0055] (1) Cell plating: the human dermal hair papilla cells in the logarithmic growth phase were adjusted to a cell density of 5x10 4 6 / mL, 100 μL / well were inoculated into 96-well plates (the edge wells were filled with sterile PBS), and incubated at 37°C, 5% CO2.

[0056] (2) Grouped administration: After the cells adhere, replace the culture medium containing different concentration gradients of Platycladus orientalis leaf alcohol extract or Platycladus orientalis leaf water extract for culture, and set up a blank control group (Control) and a positive control group (Minoxidil) respectively, 7 replicate wells for each treatment, and the positive control group uses minoxidil.

[0057] (3) CCK8 detection: After 48 h of drug action, the supernatant is aspirated, 110 μL of diluted CCK8 working solution is added per well, the culture plate is gently shaken several times, and 37°C, 5% CO2 constant temperature incubation is continued for 2 h, the absorbance value of each well is measured at 450 nm, and the cell proliferation rate is calculated.

[0058] 3, Effect of Platycladus orientalis leaf alcohol extract on mouse hair growth

[0059] C57BL / 6 mice are used to establish a hair loss animal model. Mouse hair growth includes a resting period, a regression period and a growth period, which is called a hair growth cycle. The depilated skin begins to darken as the hair follicle transitions from the resting phase to the growth phase. C57BL / 6 mice contain melanocytes in the hair follicle, which can synthesize a large amount of melanin when in the growth phase, and the skin is black. In the regression phase, melanin synthesis decreases, and the skin is gray. In the resting phase, melanin is not synthesized, and the skin is pink. The hair follicles of 6-8 week old C57BL / 6 mice enter the resting phase, so hair growth is slow during this period, and the back skin is black. After administration, the color of the mouse back skin and the hair coverage can be used to determine whether the drug promotes the mouse hair to enter the growth phase.

[0060] (1) Experimental grouping

[0061] C57BL / 6 (SPF level) male mice are randomly divided into 4 groups, 6 mice in each group, and are stored in separate cages and numbered (blank group, model group, minoxidil group, Platycladus orientalis group).

[0062] (2) Modeling

[0063] After 7 days of adaptive feeding, when the mice enter the hair resting phase, except for the blank group, the other groups of C57BL / 6 mice are injected intraperitoneally with chloral hydrate (4%, w / v) anesthesia, and the back hair is removed with a razor, leaving a 2*2 cm blank area, to establish the model.

[0064] (3) Drug administration

[0065] After the model is established, 0.32 ml of the corresponding Platycladus orientalis alcohol extract is applied to the back of the mouse, which is done once a day for 20 days. After administration, the mouse back is covered with an appropriate size of plastic wrap to prevent the drug solution from flowing off the back, and the mouse is in a state of anesthesia during the application period. During the administration period, the back hair growth of each group of rats is observed and recorded every 5 days.

[0066] (4) Determine the coverage rate of new hair growth

[0067] On days 5, 10, 15, and 20 of drug administration, the hair coverage area on the back of mice was calculated using ImageJ software. Hair coverage (%) and the promoting effect (EE) were calculated using the following formulas:

[0068] ;

[0069] ;

[0070] 4. Analysis of the anti-hair loss / hair growth promotion effects of ethanol and water extracts of Platycladus orientalis leaves

[0071] (1) Analysis of the effects of ol extract and water extract of Platycladus orientalis leaf on the proliferation of human dermal papilla cells

[0072] The results are as follows Figure 1 As shown, A is the ethanol extract of Platycladus orientalis leaves, and B is the aqueous extract of Platycladus orientalis leaves. *P<0.05, **P<0.01, ***P<0.001 indicate comparison with the blank control group; ##P<0.01, ###P<0.001 indicate comparison with the positive control group.

[0073] Depend on Figure 1 It was found that the ethanol extract of Platycladus orientalis leaves significantly promoted the proliferation of dermal papilla cells, which are important dermal cells for hair follicle growth. The cell proliferation activity was strongest at 100 µg / mL, increasing by 68% compared to the blank control group. The aqueous extract of Platycladus orientalis leaves also significantly promoted the proliferation of dermal papilla cells, which are important dermal cells for hair follicle growth. The cell proliferation activity was strongest at 200 µg / mL, increasing by 66% compared to the blank control group. Compared with the aqueous extract, the ethanol extract of Platycladus orientalis leaves had a stronger ability to promote the proliferation of dermal papilla cells. Therefore, the ethanol extract of Platycladus orientalis leaves was selected for testing hair growth in mice.

[0074] (2) Analysis of the effect of Platycladus orientalis leaf extract on hair growth in mice

[0075] The results are as follows Figure 2 The figure shows the changes in hair growth on the back of mice in different groups on days 5, 10, 15, and 20 after drug administration. Table 1 shows the hair coverage on the back of mice in each group on days 5, 10, 15, and 20.

[0076] Depend on Figure 2It can be seen that after the 5th day of administration, the hair on the back of the mice in each group did not grow significantly, but the skin color changed; after the 10th day, hair growth tended to be obvious, and the hair of mice in different groups grew to different degrees, among which the promoting effect of the Platycladus orientalis group was most obvious, indicating that Platycladus orientalis can significantly shorten the transition time of hair follicles from the resting phase to the growth phase and the time from entering the growth phase to the hair growing out of the skin. After 20 days, compared with the model group, the hair on the back of the mice in each group grew out, and the hair growth of the Platycladus orientalis group was comparable to that of the positive group of minoxidil. Therefore, the Platycladus orientalis extract has a promoting effect on hair growth.

[0077] As can be seen from Table 1, on the 5th day of administration, the shaved areas of the positive group and the Platycladus orientalis group did not grow hair. On the 10th day of administration, the skin color on the back of the mice in the Platycladus orientalis group changed, and on the 15th day, the back of the mice in the positive drug group began to turn black. On the 20th day, the back of the mice in each group was mostly covered with hair. Among them, in terms of the coverage area of the new hair, the Platycladus orientalis group had a better effect on promoting hair growth. The percentage of hair coverage was calculated based on the ratio of the coverage area of the new hair to the total shaved area of the mice in each group, and the ratio of the percentage of hair coverage of the mice in each administration group to the percentage of hair coverage of the mice in the model group on the 12th day (EE) was used to evaluate the promoting effect of each drug on hair recovery. The Platycladus orientalis group > the positive group (minoxidil group).

[0078] Table 1

[0079]

[0080] Example 2 Analysis of components of Platycladus orientalis alcohol extract and Platycladus orientalis water extract

[0081] 1. Liquid chromatography-mass spectrometry analysis method of Platycladus orientalis alcohol extract and water extract

[0082] Take 100 μL of the test solution and centrifuge at 4°C for 10 min at a centrifugal force of 20000 xg. Take the upper clear liquid and analyze it on the machine. The mass spectrometry conditions are shown in Table 2, the chromatography conditions are shown in Table 3, and the detection conditions are shown in Table 4.

[0083] The data collected by high-resolution liquid chromatography is processed by CD 3.3 (Compound Discoverer 3.3) (ThermoFisher) to complete the preliminary arrangement of the data, and then the database (mzCloud) is searched and compared, and the components are attributed and classified by combining literature analysis.

[0084] Table 2

[0085]

[0086] Table 3

[0087]

[0088] Table 4

[0089]

[0090] 2. Analysis of test results

[0091] Figure 3 and Figure 4 are the positive and negative ion flow diagrams of the alcohol extract of Platycladus orientalis leaves; Figure 5 and Figure 6 are the positive and negative ion flow diagrams of the water extract of Platycladus orientalis leaves; Table 5 is the top six flavonoids in the alcohol extract and water extract of Platycladus orientalis leaves.

[0092] As can be seen from Table 5, the top four flavonoids commonly found in the alcohol extract and water extract of Platycladus orientalis leaves are myricetin, catechin, aforoside, and tricetin; the contents of the four compounds in the alcohol extract are higher than those in the water extract; the content ratio of myricetin and tricetin in the alcohol extract and water extract of Platycladus orientalis leaves is about 6.1:1 (alcohol extract) and 12.5:1 (water extract).

[0093] Table 5

[0094]

[0095] Example 3 Network pharmacology analysis of alcohol extract and water extract of Platycladus orientalis leaves

[0096] 1. Network pharmacology method of Platycladus orientalis leaf extract

[0097] (1) Active ingredient screening and active ingredient target prediction of Platycladus orientalis leaf extract

[0098] Using Platycladus orientalis and its Latin name as the search term, all chemical components were searched through TCMSP, and chemical components that simultaneously met the molecular weight <500 molar mass and drug-likeness (DL) ≥0.18 were screened. The active ingredients of the alcohol extract and water extract of Platycladus orientalis obtained by liquid chromatography-mass spectrometry were used as potential active ingredients, and active compounds not predicted were supplemented by combining with literature reports to construct an active ingredient dataset of Platycladus orientalis.

[0099] Through the Pubchem database, the 3D structure and SMILES format of the active ingredients of Platycladus orientalis were obtained, and then imported into the Swisstargetprediction database to obtain the corresponding predicted targets, and the related targets with Probability > 0 were selected. The target name was converted to a unified gene name using the Uniport database, and the species was limited to Homo sapiens.

[0100] (2) Prediction of alopecia disease targets

[0101] The keywords "alopecia" and its synonym "hair loss" were input into the GeneCard, OMIM, TTD, and database to retrieve genes related to hair loss. The target points with a score greater than 10 in the GeneCard database were screened, and all disease targets in the OMIM and TTD databases were obtained. The results were integrated to remove duplicates to obtain the target points of hair loss diseases.

[0102] (3) Construction of PPI network of active components of Platycladus orientalis and hair loss targets and screening of core components

[0103] The target genes of the active components of Platycladus orientalis were intersected with the hair loss-related target genes, and a Venn diagram was drawn to obtain the common gene targets of Platycladus orientalis for treating hair loss. The intersection targets screened by mapping were uploaded to the String database, "Homo sapiens" was selected as the research species, and the "tsv" format file of protein-protein interaction relationships was downloaded. Subsequently, the results were imported into Cytoscape 3.10.0 software for visualization and analysis, and then the CytoNCA plug-in tool was used for topological analysis. According to the "Degree" value in the software analysis results, the core components of the drug were screened.

[0104] The intersection targets screened by mapping were uploaded to the String database, "Homo sapiens" was selected as the research species, and the "tsv" format file of protein-protein interaction relationships was downloaded. Subsequently, the results were imported into Cytoscape 3.10.0 software for visualization and analysis, and then the Centiscape plug-in tool was used for analysis, and according to the "Betweenness", "Closeness", and "Degree" values in the software analysis results, the key core target points of Platycladus orientalis extract for preventing hair loss and promoting hair growth were screened.

[0105] 2. Network pharmacology results analysis of Platycladus orientalis extract

[0106] (1) Active component screening and active component target prediction of Platycladus orientalis extract

[0107] Liquid chromatography-mass spectrometry analysis resulted in 57 active components, supplemented by 6 missing active components from related literature, totaling 63 active components. The action targets of these 63 active components were combined and duplicates were deleted, resulting in 630 potential active component action targets of Platycladus orientalis.

[0108] (2) Prediction of hair loss disease targets

[0109] The target points related to hair loss were collected from Genecards, OMIM, and TTD databases, and the repeated values were deleted to finally obtain 1648 target points related to hair loss.

[0110] The screened active ingredient target point set of Platycladus orientalis and the target point set of hair loss disease were drawn into a Venn diagram, as shown in Figure 7 The common target points of Platycladus orientalis and hair loss were obtained, and there were 124 common target points. These overlapping genes are important genes for promoting hair growth of Platycladus orientalis.

[0111] (3) PPI network construction of active ingredients of Platycladus orientalis and hair loss target points and screening of core ingredients

[0112] By using Cytoscape3.10.0 software, the interaction network of “drug-active ingredient-target-disease” was further constructed. Further analysis of the obtained network showed that it contained 124 nodes and 1726 edges, and each edge represented the relationship between Platycladus orientalis and active ingredients, active ingredients and target points, and target points and diseases. After sorting the compounds according to Degree (Degree>20), it was found that myricetin and quercetin were closely related to the target points of hair loss disease, and the content of myricetin and quercetin in Platycladus orientalis extract was the most. Therefore, myricetin and quercetin played a dominant role in the process of treating hair loss of Platycladus orientalis.

[0113] The target protein interaction network is shown in Figure 8 and Table 6, and 26 potential core target points of Platycladus orientalis extract for treating hair loss / hair growth promotion were screened, which were STAT3, TP53, ERBB2, EGFR, BCL2, AKT1, CTNNB1, MAPK1, IL2, CCND1, EP300, APP, ALB, MTOR, JUN, TNF, AR, ESR1, SRC, SNCA, ACE, PPARG, CXCL8, ESR2, SHH, CYP19A1.

[0114] Based on the above result analysis, it can be concluded that the effect of Platycladus orientalis ethanol extract on preventing hair loss and promoting hair growth is better than that of Platycladus orientalis water extract, and the active ingredient may be flavonoids; combined with the results of liquid chromatography and network medicine, it can be known that myricetin and quercetin may play a dominant role in the process of treating hair loss of Platycladus orientalis extract (mainly ethanol extract); the ratio of myricetin and quercetin is 6.1:1 (Platycladus orientalis ethanol extract) or 12.5:1 (Platycladus orientalis water extract).

[0115] Table 6 Core target points of active ingredients of Platycladus orientalis extract for preventing hair loss and promoting hair growth

[0116]

[0117] Example 4 Study on synergistic anti-hair loss and hair growth promotion effects of myricetin and quercetin

[0118] 1. Study on anti-hair loss and hair growth promotion effects of different compositions of myricetin and quercetin (5α-reductase inhibition rate)

[0119] (1) Preparation of experimental samples

[0120] Table 7 Weight ratio of myricetin and quercetin compositions

[0121]

[0122] Each composition was prepared according to the formulation amount shown in Table 7 according to the following steps:

[0123] S1 An appropriate amount of myricetin and quercetin was weighed and dissolved in anhydrous ethanol to prepare stock solutions with concentrations of 0.3%, 0.03%, and 0.003%.

[0124] S2 Compositions with concentrations of 0.3%, 0.03%, and 0.003% were prepared, and the total concentration of each concentration composition was unchanged. The proportion of each component was adjusted according to the formulation amount shown in Table 7.

[0125] (2) 5α-reductase inhibition rate test

[0126] 2-1 Stimulation of standard enzyme by test sample

[0127] A. Prepare the standardized enzyme and dilute it to a concentration of 80 U / L, and plate it in a cell culture plate. Add 100 μL of volume to each well.

[0128] B. Prepare samples (compositions 1-8 and comparative examples 1-2). According to the pre-experiment, configure the concentration points of the test sample concentration to stimulate the standard enzyme. The blank control is stimulated with sterile enzyme-free water, and the stimulation concentration is 100 μL. The time is 6 h, then transfer the supernatant to an EP tube, centrifuge to take the supernatant, and wait for detection.

[0129] C. Enzyme amount detection kit.

[0130] 2-2 Kit detection

[0131] A. Sample addition: accurately add 50 μL of standard and sample to the enzyme-labeled coated plate. Add the sample to the bottom of the enzyme-labeled plate hole, try not to touch the hole wall, and gently shake to mix.

[0132] B. Incubation: after sealing the plate with a sealing film, incubate at 37 ℃ for 30 min.

[0133] C. Liquid preparation: dilute the 30-fold concentrated washing solution with distilled water 30 times for standby use.

[0134] D. Wash: Carefully remove the plate seal, discard the liquid, spin down, fill each well with wash buffer, discard after 30 seconds, repeat 5 times, tap dry.

[0135] E. Add enzyme: Add 50 μL of enzyme reagent to each well, except the blank wells.

[0136] F. Incubate: Operate as in B.

[0137] G. Wash: Operate as in D.

[0138] H. Color development: Add 50 μL of color reagent A to each well, then add 50 μL of color reagent B, mix gently, color development at 37°C for 15 minutes in the dark.

[0139] I. Stop: Add 50 μL of stop solution to each well, stop the reaction (at this time the blue color turns to yellow).

[0140] J. Measure: Take the absorbance (OD value) of each well at 450 nm wavelength, with the blank as zero. The measurement should be performed within 15 minutes after adding the stop solution.

[0141] 2-3 Calculation formula of the enzyme activity of the sample to be tested is as follows:

[0142] × 100%.

[0143] 2, Myricetin and different compositions of five hydroxy flavone anti-hair loss and hair growth promoting effect research results

[0144] Table 8 Inhibition rate of myricetin and different compositions of five hydroxy flavone on 5α-reductase

[0145]

[0146] Table 8 shows the inhibition rate of myricetin and different compositions of five hydroxy flavone on 5α-reductase. The results show that: when the ratio of myricetin to five hydroxy flavone is 1-14:1, the composition has a synergistic effect, and the inhibition rate of the composition on 5α-reductase is better than that of myricetin (comparative example 1) and five hydroxy flavone (comparative example 2) alone; and when the ratio of myricetin to five hydroxy flavone is 1-10:1, the higher the content of myricetin in the composition, the higher the inhibition rate of the composition on 5α-reductase.

[0147] Finally, it should be pointed out that the above embodiments are only used to illustrate the technical solutions of the present application, and are not limited thereto; although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or make equivalent substitutions for part or all of the technical features; and these modifications or substitutions do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application.

Claims

1. Use of a composition in the preparation of a product for promoting hair growth and / or preventing hair loss, characterized in that, the composition consists of myricetin and tricetin; the weight ratio of the myricetin and tricetin is 1-14:

1.

2. Use according to claim 1, characterized in that, the promotion of hair growth and / or the prevention of hair loss is in the form of inhibition of 5α-reductase.

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