Natural monomer composition and application thereof in hair-blacking products

Abstract

The invention provides a natural monomer composition and application thereof in hair-blacking products, and belongs to the technical field of hair blacking. The composition comprises naringenin and luteolin in a mass ratio of (10: 1)-(1: 10), the two components have an obvious synergistic effect on hair blackening, and the composition is suitable for being prepared into a natural monomer product which is simple in formula, exact in effective component, safe and effective and has the hair blackening effect and has a wide market prospect.

Description

Technical Field

[0001] The invention belongs to the technical field of hair darkening, and particularly relates to a natural monomer composition and application thereof in hair darkening products. Background Art

[0002] Thick black hair has always been considered a symbol of good health and energy. The Suwen · Six Sections on Zang-Xiang states: "The kidney is responsible for hibernation, the root of storage, the place where the essence resides, and its beauty is in the hair." The Zhubingyuanhou Lun · Hair Diseases states: "If the kidney qi is weak, the bone marrow will be exhausted, so the hair will turn white." From this, we can see that ancient Chinese medicine believed that thick black hair represented sufficient kidney qi and vigorous qi and blood.

[0004] Studies have found that the leaves of Candelabra have the potential to darken and strengthen hair. For example, Chinese patents CN110279801A and CN102908288A respectively provide Chinese medicine formulas for darkening hair containing Candelabra leaves. However, the darkening hair effect of natural monomer substances in Candelabra leaves still needs further research. Chinese patent CN114948940A provides a natural product monomer composition with the function of darkening and strengthening hair, which is a combination of red pine lactone and quercetin. It is found that when the active ingredient red pine lactone from Platycladus orientalis leaf extract and the active ingredient quercetin from the leaf extract of Cistanche deserticola are used together, their efficacy is better than when the two are used alone. However, the darkening hair effect of the patented composition still needs to be improved. Summary of the invention

[0005] In order to solve the above problems, the inventors of the present application screened out several natural monomer substances in the leaves of Candida altissima after molecular docking in the early stage and combined them for experiments. They found that the natural monomer substances luteolin and naringenin in the leaves of Candida altissima can significantly increase the melanin content in the skin and hair of mice with leukoderma after mixing, and make the color of white hair darker or even black. Based on this, the present application proposes a natural monomer composition with high safety, clear effective ingredients and significant effect of improving black hair.

[0006] On the one hand, the present application provides a natural monomer composition with the effect of blackening hair, wherein the composition comprises naringenin and luteolin, wherein the mass ratio of naringenin to luteolin is 10:1 to 1:10.

[0007] In one embodiment, the mass ratio of naringenin to luteolin is 5:1 to 1:5.

[0008] Preferably, the mass ratio of naringenin to luteolin is 1:1, 2:1, 3:1, 4:1, 5:1, 1:2, 1:3, 1:4, 1:5, more preferably 1:1, 3:1, 5:1 and 1:5, more preferably 3:1.

[0009] In one embodiment, the natural monomer composition can be obtained by weighing and directly mixing in parts by mass within the above mass ratio range.

[0010] In one embodiment, the naringenin and luteolin are derived from artificial synthesis or plant extraction.

[0011] Optionally, the artificial synthesis can be carried out using known chemical synthesis routes of naringenin and / or luteolin, as well as known synthesis methods.

[0012] Optionally, the plant is selected from Candlewood or Citrus plants, preferably Candlewood, and more preferably Candlewood leaves.

[0013] Optionally, the plant extraction method can be a method known in the art, such as reflux extraction with water or an organic solvent, ultrasonic extraction, microwave extraction, etc.

[0014] On the other hand, the present application provides the use of the natural monomer composition in the preparation of tyrosinase activating reagents, melanin synthesis promoting reagents and / or products for treating gray hair.

[0015] In one embodiment, the melanin synthesis stimulating agent is administered to B16 melanoma cells.

[0016] On the other hand, the present application provides a drug for treating gray hair, comprising the natural monomer composition.

[0017] On the other hand, the present application provides a tyrosinase activating reagent comprising the natural monomer composition.

[0018] On the other hand, the present application provides a melanin synthesis promoting agent, comprising the natural monomer composition.

[0019] In one embodiment, the content of the natural monomer composition in the drug for treating gray hair, the tyrosinase activating agent or the melanin synthesis promoting agent is 0.01%-99.99%, where % is the mass volume percentage (w / v) or the mass percentage (w / w).

[0020] On the other hand, the present application provides the use of the natural monomer composition in preparing a product with hair darkening effect.

[0021] In one embodiment, the product is selected from medicines, health products, functional foods, special medical foods, cosmetics or daily chemical products.

[0022] In one embodiment, the dosage form of the product is selected from oral preparations, suppositories, aerosols, drops, injections or external preparations.

[0023] Optionally, the oral preparation includes but is not limited to oral liquid, suspension, granules, capsules, powders, tablets, pills, etc.

[0024] Optionally, the injection method of the injection includes but is not limited to intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, etc.

[0025] Optionally, the topical agent includes but is not limited to gel, ointment, cream, foam, shampoo, conditioner, etc.

[0026] In one embodiment, the content of the natural monomer composition in the product is 0.01%-99.99%, where % refers to mass volume percentage (w / v) or mass percentage (w / w).

[0027] In one embodiment, the natural monomer composition in the product is prepared into a compound preparation with the same active ingredient or other active ingredients, or further contains at least one pharmaceutically acceptable excipient.

[0028] In one embodiment, the ingredients with the same activity include but are not limited to extracts or chemicals with hair blackening activity from Platycladus orientalis, Polygonum multiflorum, black sesame, mulberry, etc.

[0029] In one embodiment, the other active ingredients include, but are not limited to, anti-inflammatory, antioxidant, anti-allergic, uric acid-lowering, anti-tumor, antibacterial, antiviral and other active ingredients.

[0030] In one embodiment, the pharmaceutically acceptable excipients include, but are not limited to, water, organic solvents, preservatives, thickeners, surfactants, fillers, disintegrants, and the like.

[0031] It is to be understood that the “hair” mentioned in the present application refers to the hair of animals, including but not limited to humans or mice; the hair includes but is not limited to the head or hair from other parts of the body.

[0032] Vaccinium bracteatum Thunb., also known as the black rice tree, is a plant of the genus Vaccinium in the family Ericaceae. Vaccinium berries, roots and leaves can all be used as medicine. Vaccinium berries have the effects of nourishing the liver and kidneys, strengthening the tendons and bones, consolidating the essence and qi, and stopping diarrhea. Vaccinium roots can be used fresh or dried, and can disperse blood stasis, relieve pain, and treat toothache and swelling and pain caused by falls. Vaccinium leaves are the edible part of Vaccinium bracteatum. They are mild in nature and slightly sour and astringent. The chemical composition of the leaves is complex, containing a variety of fat-soluble active ingredients such as hexadecane, terpenoids, terpenoid alcohol, quercetin, and isoorientin. There is a long history of picking Vaccinium bracteatum leaves around the Qingming Festival in spring and making juice to make black rice for consumption. "Ben Cao Jing Shu" says: "Vaccinium bracteatum, together with Eclipta prostrata, Gallic acid, Rehmannia root, Morus alba, Lycium barbarum, Cornus officinalis, Polygonum multiflorum, and Tribulus terrestris, are holy medicines for black hair and beards. They have a peaceful smell and are non-toxic."

[0033] Luteolin, chemically known as 3',4',5,7-tetrahydroxyflavone, is a flavonoid found in the leaves of Candlewood. Its chemical formula is C 15 H 10 O6, molecular weight 286.23. Luteolin is a natural antioxidant with strong anti-inflammatory, anti-diabetic, and cardioprotective functions. Luteolin can scavenge free radicals, protect cells, and interact with other antioxidants such as vitamins to enhance antioxidant effects. Luteolin can inhibit pro-inflammatory mediators such as NO, TNF-α, IL-6, and IL-β, while inhibiting inflammatory signal transduction pathways such as NF-κB, AP-1, and JAK-STAT pathways. Luteolin can also absorb ultraviolet rays, exert antioxidant and anti-inflammatory effects to protect human keratinocytes and fibroblasts, and ultimately relieve skin aging and inhibit dermatitis. Luteolin also has significant anti-cancer effects and can treat colon cancer and breast cancer through multiple signal pathways.

[0034] The chemical name of naringenin is 4',5,7-trihydroxyflavone, and its chemical formula is C 15 H 12 O5, with a molecular weight of 272.26, is a common structure that shares two aromatic rings, connected by a linear 3-carbon chain (C6-C3-C6) to form an oxygen-containing heterocycle. It is a flavanone extracted from flavonoids and belongs to the flavonoid class of compounds. It is mainly found in citrus fruits such as grapes, oranges, and lemons. Naringenin has a protective effect on the heart, participates in the regulation of the cardiovascular system through multiple signaling pathways, and slows down the progression of cardiovascular diseases. Naringenin also has anti-inflammatory, antibacterial, immune-regulating, free radical scavenging and antioxidant effects.

[0035] This application has at least the following beneficial effects:

[0036] 1. After molecular docking screening and experiments on natural product monomer compounds from the leaves of Candlewood, the present application unexpectedly found that although they are all flavonoid compounds from the same plant source and have similar structures, different natural monomer combinations have huge differences in the effects of activating tyrosinase, activating melanoma cells to synthesize melanin, and treating mice with leucophyllum. This discovery plays a positive role in the study of the pharmacological activity of monomer substances and their combinations in Candlewood leaf extracts. Among them, luteolin itself has a weak ability to blacken hair, but when naringenin and luteolin are combined in a specific mass ratio, a significantly improved tyrosinase activation effect is obtained, and it has a significant effect on the treatment of mice with leucophyllum in animal experiments, and there is an obvious synergistic effect.

[0037] 2. The natural monomer composition provided in this application has natural ingredients, high safety, simple formula, and can be used to prepare products in various fields such as food, medicine, daily chemical products, etc., and has broad market prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the invention and, together with the description, serve to explain the principles of the invention.

[0039] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following briefly introduces the drawings required for use in the embodiments or the prior art descriptions. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work. In the drawings:

[0040] Figure 1 1 is a result diagram of the change in the hair color on the back of mice in Test Example 3, wherein a to f are result diagrams of the blank group, the luteolin group, the Example 1 group, the Example 2 group, the Example 3 group and the naringenin group, respectively;

[0041] Figure 2 3 is the observation result diagram of the hair structure of mice in Test Example 3, wherein a to f are the result diagrams of the blank group, the luteolin group, the Example 1 group, the Example 2 group, the Example 3 group and the naringenin group, respectively. DETAILED DESCRIPTION

[0042] In order to more clearly explain the overall concept of the application, the following is described in detail by way of embodiment. In the following description, a large amount of specific details are provided so as to provide a more thorough understanding of the application. However, it is obvious to those skilled in the art that the application can be implemented without one or more of these details. In other examples, in order to avoid confusion with the application, some technical features well known in the art are not described.

[0043] Unless otherwise specified, in the following embodiments, reagents or instruments used without indicating the manufacturer are all conventional products that can be purchased from the market. If no specific conditions are indicated in the examples, conventional conditions or conditions recommended by the manufacturer are followed.

[0044] The natural monomers luteolin and naringenin in the present application can be extracted and separated from the leaves of Candlewood and pomelo plants containing these active ingredients by conventional extraction and purification methods in the chemical field, or can be purchased from commercial products.

[0045] The luteolin and naringenin in the following examples are all commercially available products, wherein the CAS number of luteolin is: 491-70-3; the CAS number of quercetin is: 117-39-5; mice were provided by Beijing Weitonglihua, and B16 melanoma cells were provided by Wuhan Pronocell Life Science Co., Ltd.

[0046] Example 1

[0047] This embodiment provides a composition of natural monomer components, consisting of naringenin and luteolin, and the mass ratio of the two components is 1:1.

[0048] Embodiments 2 to 4

[0049] Examples 2 to 4 respectively provide a composition of a natural monomer component, wherein all components in the composition are the same as those in Example 1, namely, composed of naringenin and luteolin, except that the mass ratios of the two components are 3:1, 5:1 and 1:5, respectively.

[0050] Example 5

[0051] This embodiment is an application example of the aforementioned embodiment. In one embodiment, the natural monomer composition composed of naringenin and luteolin provided in the aforementioned embodiment can be made into various products with hair blackening effects according to actual needs, such as medicines, foods, health products, special medical foods, daily chemical products, etc., wherein the content of the natural monomer composition in the above-mentioned products can be 0.01% to 99.99% (the percentage "%" can be w / w or w / v), and when the product is made, conventional excipients of the product can be added, such as water (applicable to medicines, foods, health products, special medical foods, etc.) or hair care product base (applicable to daily chemical products), or it can be compounded with other active ingredients with hair blackening effects or ingredients with other activities, and then conventional excipients of the corresponding product can be added.

[0052] The following is an activity test of the hair darkening effect of the natural monomer composition provided in the above Examples 1 to 4. In each test example, the above composition can be prepared into a test object suitable for the test.

[0053] Test Example 1: Effect of activating tyrosinase

[0054] Experimental methods:

[0055] A 100U / mL tyrosinase solution, a levodopa solution (1 mg / mL) and a sample solution with component concentrations shown in Table 1 were prepared using a phosphate buffer solution of pH 6.8. The sample solution was divided into a sample group A (20 μL of the sample to be tested + 10 μL of tyrosinase solution), a sample control group B (20 μL of the sample to be tested + 10 μL of PBS), a blank group C (20 μL of PBS + 10 μL of tyrosinase), and a blank control group D (30 μL of PBS), all of which were placed in a 96-well plate, incubated at 37°C for 10 min, 40 μL of levodopa solution was added, incubated at 37°C for 5 min, and the absorbance was measured at a wavelength of 475 nm.

[0056] Each experiment was repeated 3 times, and the absorbance of each experiment was the average value of the 3 tests.

[0057] The absorbance measured in each group is recorded as the English letter of the group, such as the absorbance measured in sample group A is recorded as A, the absorbance measured in sample control group B is recorded as B, and so on. The calculation formula is:

[0058] Tyrosinase activation rate (%) = [(AB) - (CD)] / (CD) × 100%.

[0059] The results are shown in Table 1.

[0060] Table 1 Tyrosinase activation experimental results

[0061] Group Tyrosinase activation rate (%) Naringenin 0.1mg / mL 70.36 Naringenin 0.3mg / mL 130.11 Luteolin 0.1mg / mL 11.16 Quercetin 0.1mg / mL 31.99 Naringenin 0.1mg / mL, Luteolin 0.1mg / mL 165.44 Naringenin 0.3mg / mL, Luteolin 0.1mg / mL 198.56 Naringenin 0.5mg / mL, Luteolin 0.1mg / mL 185.64 Naringenin 0.1mg / mL, Luteolin 0.5mg / mL 146.38 Naringenin 0.1mg / mL, Quercetin 0.1mg / mL 94.06

[0062] From the results in Table 1, it can be seen that the natural monomer compositions provided in Examples 1 to 4 can significantly activate the activity of tyrosinase and have a good hair darkening effect.

[0063] Moreover, when the mass ratio of naringenin to luteolin was 3:1, the tyrosinase activation rate was as high as 198.56%, which was significantly higher than that of the 0.3 mg / mL naringenin group and the 0.1 mg / mL luteolin group used alone. When the mass ratio of naringenin to luteolin was 5:1, the tyrosinase activation rate showed a downward trend, indicating that a smaller amount of naringenin and luteolin combination can achieve a better hair darkening effect.

[0064] At the same time, by comparing the data results of the naringenin and luteolin groups with mass ratios of 1:1 and 1:5 respectively, it can be seen that although luteolin itself has almost no hair blackening effect, an appropriate amount of luteolin can significantly enhance the hair blackening effect of naringenin.

[0065] Furthermore, by comparing the data results of the naringenin + luteolin group and the naringenin + quercetin group with a mass ratio of 3:1, it can be seen that although quercetin has a better activation effect on tyrosinase than luteolin, and it is known in the prior art that it has an auxiliary effect on some natural monomer substances with hair blackening effects (see patent CN114948940A), it has almost no obvious improvement effect on hair blackening after being combined with naringenin, indicating that the hair blackening effect can be significantly improved only when luteolin and naringenin are combined in an appropriate proportion.

[0066] Test Example 2: Activation effect on melanin synthesis in B16 melanoma cells

[0067] Experimental methods:

[0068] B16-F10 cells (5×10 5 / mL) were inoculated into a culture dish containing 10mL of culture medium and incubated for 24 hours. The cells were then treated with a solution of the concentration components shown in Table 2 for 48 hours. After treatment, the melanin production activity was estimated based on the amount of melanin (extracellular melanin) secreted into the culture medium. The cells were incubated in trypsin (2.5%) to separate the adherent cells. The cells were then placed in a test tube and dissolved in 1mL Triton (0.1%). The absorbance of the extracellular melanin content was measured at 475nm.

[0069] Each group of experiments was repeated 3 times, and the absorbance of each group of experiments was the average of the 3 tests. The results are shown in Table 2:

[0070] Table 2 Experimental results of activating melanin synthesis in melanoma cells

[0071]

[0072]

[0073] From the results in Table 2, it can be seen that compared with the blank control group, the natural monomer composition of naringenin and luteolin provided in the embodiment has a significantly improved melanin content, and is higher than the naringenin group and luteolin group used alone, indicating that the appropriate amount of naringenin and luteolin combination has a significant synergistic effect in improving the melanin synthesis effect of melanoma cells. In particular, when the mass ratio of naringenin to luteolin is 3:1, the melanin content synthesized by b16 melanoma cells is the highest, which is 45.92μg / mL, and the activation effect on tyrosinase in melanoma cells is the best.

[0074] Test Example 3: Effect of blackening hair on mice with leukoderma

[0075] Experimental methods:

[0076] C57BL / 6J male mice were adaptively raised for 7 days after arriving at the laboratory. They were randomly divided into 6 groups, each with 12 mice. The first group was a blank group, denoted as a, and the second to sixth groups were luteolin group, Example 1 group (naringenin: luteolin = 1:1 (w / w)), Example 2 group (naringenin: luteolin = 3:1 (w / w)), Example 3 group (naringenin: luteolin = 5:1 (w / w)) and naringenin group, denoted as b, c, d, e and f, respectively. The mice were depilated, the hair on the back was cut short with a shaver, the depilatory cream was applied to the back of the mouse for 2-3 minutes, the mouse was washed with warm water, and the depilatory area was wiped with absorbent cotton to avoid damaging the mouse skin and to ensure that there was no damage and residual hair in the depilatory area as much as possible. The depilatory area was about 2cm×2cm. Four days after hair removal, the model was established. 0.5 ml of 4% hydroquinone ointment (1 g of sodium carboxymethyl cellulose and 4 g of hydroquinone dissolved in 100 mL of deionized water) was applied to the test area on the back of the mouse. The blank control group was applied with a mixed solution of blank sodium carboxymethyl cellulose for 40 consecutive days, once a day. On the 11th day of modeling, each group of mice was given a dose of 100 mg / kg / d, and the concentration of the drug was 15 mg / ml. The blank group was gavaged with an equal amount of saline solution. The drug was administered continuously for 30 days. 2 hours after the last administration, the eyeballs were removed to collect blood, the serum was separated, and the spleen, kidneys, liver, and heart were dissected and weighed. The skin of the central part of the back test area, about 1 cm × 1 cm, was collected and fixed in tissue fluid fixative.

[0077] Among them, the color photos of the mouse hair after the experiment are as follows Figure 1 shown.

[0078] Depend on Figure 1 The results show that the hair of mice turned white significantly after modeling, and the synthesis of melanin was blocked, indicating that the modeling was successful. Among them, the hair on the back of mice in the luteolin group did not darken, indicating that luteolin was generally effective in treating gray hair; the naringenin group had a certain effect on the treatment of gray hair, the color of the back hair darkened, some gray hair returned to black hair, and most of the gray hair turned brown, but the color stopped changing after turning brown and did not turn black. The results of the embodiment group showed that the combined use of naringenin and luteolin was more effective in treating gray hair than the use of luteolin or naringenin alone, with more black area and less brown area.

[0079] The melanin content of the mouse hair after the above experiment was observed structurally. The experimental process is as follows:

[0080] Place a drop of water on the slide and add a certain amount of mouse hair, trying to separate the hair without overlapping. Cover with a cover slip, ensuring there are no bubbles between the cover slip and the slide, and observe the hair sample using a polarizing microscope with a deflection angle of 180 degrees. The results are as follows: Figure 2 shown.

[0081] Depend on Figure 2The results show that after hydroquinone modeling, the melanin content in the mouse hair was significantly reduced, and eventually the hair turned white, indicating that the modeling was successful.

[0082] Among them, luteolin and naringenin can restore the gray hair formed by hydroquinone modeling to a certain extent. The melanin content in the hair increased to a certain extent, and the brown color was more obvious, but it still did not return to the level of the blank group. As shown in the results of the implementation group, the mixture of luteolin and naringenin at a specific mass ratio has a good promoting effect on restoring the melanin level of gray hair.

[0083] The above is only an embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the scope of the claims of the present application.

Claims

1. A natural monomer composition having the effect of darkening hair, characterized in that: The composition comprises naringenin and luteolin, wherein the mass ratio of the naringenin to the luteolin is 10:1 to 1:

10.

2. The natural monomer composition according to claim 1, characterized in that The mass ratio of naringenin to luteolin is 5:1 to 1:5, preferably 1:1, 3:1, 5:1 and 1:5, and more preferably 3:

1.

3. The natural monomer composition according to claim 1, characterized in that The naringenin and luteolin are derived from artificial synthesis or plant extraction.

4. Use of the natural monomer composition according to any one of claims 1 to 3 in the preparation of a tyrosinase activating agent, a melanin synthesis promoting agent and / or a product for treating white hair.

5. Use of the natural monomer composition as described in any one of claims 1 to 3 in the preparation of a product with hair darkening effect.

6. The use according to claim 5, characterized in that: The product is selected from medicines, health products, functional foods, special medical foods, cosmetics or daily chemical products.

7. The use according to claim 5, characterized in that: The dosage form of the product is selected from oral preparations, suppositories, aerosols, drops, injections or external preparations.

8. The use according to claim 5, characterized in that: The content of the natural monomer composition in the product is 0.01%-99.99%, wherein the % refers to the mass volume percentage (w / v) or the mass percentage (w / w).

9. The use according to claim 5, characterized in that: The natural monomer composition in the product is prepared into a compound preparation with the same active ingredient or other active ingredients, or further contains at least one pharmaceutically acceptable excipient.

10. A drug for treating white hair, characterized in that: Contains the natural monomer composition as described in any one of claims 1 to 3.

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