A natural monomer composition and its use in hair growth products

Abstract

The application provides a natural monomer composition and its application in hair blackening products, and belongs to the technical field of hair blackening. The composition comprises naringenin and luteolin in a mass ratio of 10:1-1:10. The two components have obvious synergistic effect on hair blackening, are suitable for being prepared into natural monomer products with simple formula, definite effective components, safety and effectiveness, hair blackening effect, and wide market prospect.

Description

[0001] A natural monomer composition and its application in hair blackening products TECHNICAL FIELD

[0002] The present application belongs to the technical field of hair blackening, and particularly relates to a natural monomer composition and its application in hair blackening products. BACKGROUND

[0003] Black and thick hair has always been considered as a symbol of physical health and energetic. The Suwen Six Jie Cang Xiang Lun Chapter states: "The kidney is the main hibernation, the foundation of sealing and storing, and the place of essence, and its hair is in the hair." The Zhongdeng Yuanhou Lun Chapter points out: "If the kidney qi is weak, the bone marrow will be exhausted, and the hair will turn white." Therefore, ancient Chinese medicine believes that black and thick hair is a representative of sufficient kidney qi and vigorous blood.

[0004] Research has found that the leaves of Ilicium fortunei have the potential to blacken and thicken hair. For example, Chinese patents CN110279801A and CN102908288A provide hair blackening traditional Chinese medicine formulas containing the leaves of Ilicium fortunei. However, the hair blackening effect of natural monomer substances in the leaves of Ilicium fortunei still needs further research. Chinese patent CN114948940A provides a natural product monomer composition with the functions of blackening and thickening hair, which is a combination of pinostrobin and quercetin. It is found that when the active ingredient pinostrobin from the extract of Platycladus orientalis leaves and the active ingredient quercetin from the extract of Vaccinium bracteatum leaves are used together, their efficacy is better than when they are used alone. However, the hair blackening effect of the composition of the patent still needs to be improved. SUMMARY

[0005] To solve the above problems, the present inventors have screened several natural monomer substances in the leaves of Ilicium fortunei and conducted experiments on their combination after molecular docking in the early stage. It is found that the natural monomer substances luteolin and naringenin in the leaves of Ilicium fortunei can significantly increase the melanin content in the skin and hair of mice with white hair and promote the color of white hair to become darker or even black. Based on this, the present application provides a natural monomer composition with high safety, clear effective ingredients and significant hair blackening effect.

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

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

[0008] Preferably, the mass ratio of the naringenin to the 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, and more preferably 3:1.

[0009] In one embodiment, the natural monomer composition can be directly mixed by weighing the components in the above-mentioned 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 performed using known chemical synthesis routes and methods for naringenin and / or luteolin.

[0012] Optionally, the plant is selected from the group consisting of Lindera glauca and citrus plants, preferably Lindera glauca, and more preferably Lindera glauca leaves.

[0013] Optionally, the extraction method for the plant extraction can be selected from known methods in the art, such as reflux extraction, ultrasonic extraction, microwave extraction, etc. using water or organic solvents.

[0014] In another aspect, the present application provides the use of the natural monomer composition in the preparation of a tyrosinase activating agent, a melanin synthesis promoting agent, and / or a product for treating white hair.

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

[0016] In another aspect, the present application provides a medicament for treating white hair, which contains the natural monomer composition.

[0017] In another aspect, the present application provides a tyrosinase activating agent, which contains the natural monomer composition.

[0018] In another aspect, the present application provides a melanin synthesis promoting agent, which contains the natural monomer composition.

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

[0020] In another aspect, the present application provides the use of the natural monomer composition in the preparation of a product having a hair darkening effect.

[0021] In one embodiment, the product is selected from the group consisting of a pharmaceutical product, a cosmetic product, and a daily use product.

[0022] In one embodiment, the dosage form of the product is selected from the group consisting of an oral dosage form, a suppository, an aerosol, a drop, an injection, and a topical dosage form.

[0023] Optionally, the oral dosage forms include, but are not limited to, oral liquid, suspension, granules, capsules, powders, tablets, drop pills, etc.

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

[0025] Optionally, the external use dosage form includes, but is not limited to, gel, ointment, cream, foam, shampoo, hair conditioner, etc.

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

[0027] In an embodiment, the natural monomer composition in the product is compounded with the same active or other active ingredients, or further contains at least one pharmaceutically acceptable excipient.

[0028] In an embodiment, the same active ingredient includes, but is not limited to, extracts or chemicals with hair blackening activity derived from Platycladus orientalis, Radix Polygoni Multiflori, black sesame, mulberry, etc.

[0029] In an embodiment, the other active ingredient includes, but is not limited to, anti-inflammatory, antioxidant, anti-allergic, uric acid-lowering, anti-tumor, antibacterial, anti-viral, etc.

[0030] In an embodiment, the pharmaceutically acceptable excipient includes, but is not limited to, water, organic solvent, preservative, thickening agent, surfactant, filler, disintegrant, etc.

[0031] It can be understood that the "hair" 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 hair on the head or other parts of the body.

[0032] Vaccinium bracteatum Thunb., also known as Umi, is a plant of the heather family Vaccinium. The fruit, root and leaf of Vaccinium bracteatum Thunb. can be used as medicine. The fruit has the effects of tonifying liver and kidney, strengthening bones and muscles, and astringing essence, and can treat diarrhea. The root can be used fresh or dried, and can dispel blood stasis, relieve pain, treat toothache and swelling and pain caused by injury. The leaf of Vaccinium bracteatum Thunb. is the edible part of Vaccinium bracteatum Thunb., which is slightly sour and astringent, and contains complex chemical components, including hentriacontane, acerogen, acerogenol, quercetin, isohomosaligenin and other fat-soluble active ingredients. It has a long history to make Umi rice by picking the leaf of Vaccinium bracteatum Thunb. before and after the Qingming Festival in spring. "Bencao Jing Shu" says: "Vaccinium bracteatum Thunb. is the same as Eclipta prostrata, gallnut, Rehmannia glutinosa, mulberry root, medlar, hawthorn, Tribulus terrestris, and is the holy medicine for blackening hair, with mild smell and complex nature, and non-toxic."

[0033] Luteolin, also known as 3', 4', 5, 7-tetrahydroxyflavone, is a flavonoid found in the leaves of Vitex negundo var. heterophylla. Its chemical formula is C 15 H 10 O6, and its molecular weight is 286.23. Luteolin is a natural antioxidant with strong anti-inflammatory, anti-diabetic, and heart-protecting functions. It can scavenge free radicals and protect cells. It can also 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, and inhibit inflammatory signal transduction pathways such as NF-κB, AP-1, and JAK-STAT pathways. Luteolin can also absorb ultraviolet light, play an antioxidant and anti-inflammatory role in protecting human keratinocytes and fibroblasts, and ultimately alleviate skin aging and inhibit dermatitis. Luteolin also has significant anti-cancer effects and can treat colon cancer and breast cancer through multiple signaling pathways.

[0034] Naringenin, also known as 4', 5, 7-trihydroxyflavone, has a chemical formula of C 15 H 12 O5, and a molecular weight of 272.26. It is a flavanone extracted from flavonoid substances, belonging to flavonoid compounds, and mainly exists in grape, orange, lemon, and other citrus fruits. Naringenin has a protective effect on the heart and participates in the regulation of the cardiovascular system through multiple signaling pathways, slowing the progression of cardiovascular disease. Naringenin also has anti-inflammatory, antibacterial, immune regulation, free radical scavenging, and antioxidant effects.

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

[0036] 1. The present application unexpectedly found that although both are flavonoid compounds from the same plant source and have similar structures, different natural monomer combinations have a huge difference in activating tyrosinase, activating melanin synthesis in melanoma cells, and treating mice with white hair. This discovery plays a positive role in the pharmacological activity research of monomer substances and their combinations in Vitex negundo var. heterophylla extract. Among them, luteolin itself has weak hair blackening ability, but when naringenin and luteolin are combined in a specific mass ratio, the tyrosinase activation effect is significantly improved, and the treatment of mice with white hair in animal experiments has significant efficacy, showing obvious synergistic effect.

[0037] 2. The natural monomer composition provided in the application is natural in composition, high in safety, simple in formula, and can be applied to the preparation of products in various fields such as food, medicine, daily chemical products, etc., and has a broad market prospect. BRIEF DESCRIPTION OF DRAWINGS

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

[0039] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the accompanying drawings needed to be used in the embodiments or prior art description will be briefly introduced. Those skilled in the art can also obtain other drawings according to these drawings without creative labor. In the drawings:

[0040] Figure 1 is a result graph of the change of the color of the hair on the back of the mouse in test example three, wherein a~f are the result graphs 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 is a result graph of the observation of the structure of the hair of the mouse in test example three, wherein a~f are the result graphs 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 illustrate the overall concept of the application, the following will be described in the form of examples. In the following description, a large number of specific details are given in order 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, some technical features known in the art are not described in order not to obscure the application.

[0043] Unless otherwise specified, in the following embodiments, the reagents or instruments used are not marked with the manufacturer, and are all conventional products that can be obtained by market purchase. In the examples, the specific conditions are not marked, and are carried out according to the conventional conditions or the conditions recommended by the manufacturer.

[0044] In the application, the natural monomers luteolin and naringenin can be extracted and separated from the leaves of Kleinhovia nobilis and naringen plants containing these active ingredients by using conventional extraction and purification methods in the field of chemistry, or can be purchased from market products.

[0045] Luteolin and Quercetin in the following examples are commercially available, and the CAS number of Luteolin is 491-70-3, and the CAS number of Quercetin is 117-39-5. The mice are provided by Beijing Weitong Lihua, and the B16 melanoma cells are provided by Wuhan Punoxie Life Science and Technology Co., Ltd.

[0046] Example 1

[0047] This example provides a natural monomer composition consisting of Quercetin and Luteolin, and the mass ratio of the two components is 1:1.

[0048] Examples 2-4

[0049] Examples 2-4 respectively provide a natural monomer composition, wherein all components in the composition are the same as those in Example 1, i.e., consisting of Quercetin and Luteolin, and the difference is that the mass ratio of the two components is 3:1, 5:1 and 1:5 respectively.

[0050] Example 5

[0051] This example is an application example of the foregoing examples. In one embodiment, the natural monomer composition consisting of Quercetin and Luteolin provided by the foregoing examples can be made into various products with hair blackening effect according to actual needs, such as pharmaceuticals, cosmetics, daily chemical products, etc. The content of the natural monomer composition in the above products can be 0.01% to 99.99% (the percentage “%” can be w / w or w / v), and conventional adjuvants of the product can be added when the product is made, such as water (suitable for pharmaceuticals and cosmetics) or hair care product matrix (suitable for daily chemical products), or other active ingredients with hair blackening effect or other active ingredients are compounded, and then the conventional adjuvants of the corresponding product are added.

[0052] The following natural monomer compositions provided by the foregoing Examples 1-4 are tested for hair blackening effect, and each test example can prepare the foregoing composition into a test object suitable for the test.

[0053] Test Example 1: Activation effect on tyrosinase

[0054] Experimental method:

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

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

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

[0058] The activation rate of tyrosinase (%) = [(A-B)-(C-D)] / (C-D) x 100%.

[0059] The results are shown in Table 1.

[0060] Table 1: Results of tyrosinase activation experiment

[0061]

[0062] As can be seen from the results in Table 1, the natural monomer compositions provided in Examples 1-4 can all significantly activate the activity of tyrosinase, and have good hair blackening effects.

[0063] Moreover, when the mass ratio of naringenin and luteolin is 3:1, the activation rate of tyrosinase is as high as 198.56%, which is significantly improved compared with the 0.3 mg / mL naringenin group and the 0.1 mg / mL luteolin group used alone. When the mass ratio of naringenin and luteolin is 5:1, the activation rate of tyrosinase decreases, indicating that a smaller amount of naringenin and luteolin composition can achieve a better hair blackening effect.

[0064] At the same time, by comparing the data of the naringenin and luteolin groups with mass ratios of 1:1 and 1:5, it can be seen that although luteolin itself has little hair blackening effect, an appropriate amount of luteolin can significantly enhance the hair blackening effect of naringenin.

[0065] Further, 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 known that although the activation effect of quercetin on tyrosinase is better than that of luteolin, and it is known in the prior art that the natural monomer substance (see patent CN114948940A) having a hair blackening effect has an auxiliary effect, but after being compounded with naringenin, it has almost no obvious promotion effect on hair blackening, which indicates that only in the compound combination of appropriate proportions of luteolin and naringenin, the hair blackening effect can be significantly improved.

[0066] Test Example Two: Activation Effect on Melanin Synthesis in B16 Melanoma Cells

[0067] Experimental Method:

[0068] B16-F10 cells (5x10 5 were inoculated into a culture dish containing 10 mL of culture medium and incubated for 24 hours. Then the cells were treated with a drug solution of the components at the concentrations shown in Table 2 for 48 hours. After treatment, the melanin production activity was estimated from the amount of melanin (extracellular melanin) secreted into the culture medium. The adherent cells were separated by incubating the cells in trypsin (2.5%). Then the cells were placed in a test tube and dissolved in 1 mL of Triton (0.1%). The absorbance of the extracellular melanin content was measured at 475 nm.

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

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

[0071]

[0072] As can be seen from the results of Table 2, compared with the blank control group, the natural monomer composition of naringenin and luteolin provided by the embodiment has a significantly improved melanin content, and is higher than that of the naringenin group and the luteolin group used alone, which indicates that the appropriate combination of naringenin and luteolin has obvious synergistic effect in improving the melanin synthesis effect of melanoma cells. Especially when the mass ratio of naringenin and luteolin is 3:1, the melanin content synthesized by b16 melanoma cells is the highest, which is 45.92 μg / mL, and the activation effect on tyrosinase in melanoma cells is the best.

[0073] Test Example Three: Hair Blackening Effect on White Hair Mice

[0074] Experimental Method:

[0075] C57BL / 6J public mice were adaptively fed for 7 days after arriving in the laboratory. Randomly divided into 6 groups, 12 in each group, the first group was blank group, recorded as a, the second to the sixth group 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, respectively recorded as b, c, d, e and f. The mice were depilated, the hair on the back was cut short with a shaver, and the depilatory cream was applied to the back of the mice for 2-3 min, washed with warm water, and wiped with absorbent cotton in the depilation area to avoid damaging the mouse skin and ensure that the depilation area was not damaged and had no residual hair. The depilation area was about 2 cm x 2 cm. Four days after depilation, modeling began, and 0.5 ml of 4% hydroquinone ointment (1 g of carboxymethylcellulose sodium and 4 g of hydroquinone were dissolved in 100 mL of deionized water) was applied to the test area on the back of the mice. The blank control group was applied with a blank carboxymethylcellulose sodium mixed solution, and was continuously applied for 40 days, once a day. On the 11th day of modeling, each group of mice was administered at a dose of 100 mg / kg / d, and the drug concentration was 15 mg / ml. The blank group was administered with an equal amount of normal saline solution. Continuous administration for 30 days. 2 hours after the last administration, the eyeball was removed to collect blood, the serum was separated, and the spleen, kidney, liver and heart were weighed. The center part of the skin in the test area on the back was collected about 1 cm x 1 cm and fixed in the tissue liquid fixing solution.

[0076] The color of the mouse hair after the experiment is shown in Figure 1 .

[0077] From the results in Figure 1 , it can be seen that the mouse hair after modeling is obviously white, and the synthesis of melanin is blocked, indicating that the modeling is successful. Among them, the mouse hair on the back of the luteolin group did not become darker, indicating that luteolin has better effect on treating white hair; the naringenin group has a certain effect on treating white hair, the hair color on the back becomes darker, part of the white hair returns to black hair, and most of the white hair turns to brown, but the color stops changing after turning to brown, and does not turn to black. The results of the example group show that the combination of naringenin and luteolin has better effect on treating white hair than using luteolin or naringenin alone, and has more black area and less brown area.

[0078] The melanin content of the mouse hair after the above experiment was observed, and the experimental process was as follows:

[0079] A drop of water was added to the slide, and a certain amount of mouse hair was added to separate the hair as much as possible without overlapping. Cover the cover glass to ensure that there is no air bubble between the cover glass and the slide. Use a polarizing microscope to observe the hair sample with a deflection angle of 180 degrees. The results are shown in Figure 2 .

[0080] From the results in Figure 2The results in Table 1 show that the content of melanin in the hair of the mice is significantly reduced after the hydroquinone modeling, and the hair is ultimately white, indicating that the modeling is successful.

[0081] Among them, luteolin and naringenin can restore the white hair formed by hydroquinone modeling to some extent, the content of melanin in the hair increases to a certain extent, the brown color is more obvious, and it has not yet recovered to the level of the blank group. The results of the implementation group show that the mixture of luteolin and naringenin in a certain mass ratio has a good promoting effect on restoring the melanin level of white hair.

[0082] The above only describes the embodiments of the present application and is not intended to limit the present application. Various modifications and changes can be made to the present application by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the scope of the claims of the present application.

Claims

1. A natural monomer composition having hair blackening effect, characterized in that, The composition consists of naringenin and luteolin in a mass ratio of 3:

1.

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

3. Use of the natural monomer composition according to claim 1 or 2 in the preparation of a treatment for canities.

4. Use of the natural monomer composition according to claim 1 or 2 in the preparation of a medicament or a daily-use product with a hair-darkening effect.

5. Use according to claim 4, characterized in that, The medicament is in a dosage form selected from the group consisting of oral dosage, suppository, aerosol, drops, injection or external preparation.

6. Use according to claim 4, characterized in that, The content of the natural monomer composition in the medicament or daily-use product is 0.01%-99.99%, which is mass volume percentage (w / v) or mass percentage (w / w).

7. Use according to claim 4, characterized in that, The natural monomer composition in the medicament or daily-use product is compounded with the same active ingredient or other active ingredients, or further contains at least one pharmaceutically acceptable excipient.

8. A medicament for treating poliosis, characterized by, The medicament or daily-use product contains the natural monomer composition according to claim 1 or 2.

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