CN111514048A - Natural component for treating melanin pigmentation and application of natural component in preparing freckle-removing and whitening cosmetics - Google Patents
Natural component for treating melanin pigmentation and application of natural component in preparing freckle-removing and whitening cosmetics Download PDFInfo
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- CN111514048A CN111514048A CN202010479506.0A CN202010479506A CN111514048A CN 111514048 A CN111514048 A CN 111514048A CN 202010479506 A CN202010479506 A CN 202010479506A CN 111514048 A CN111514048 A CN 111514048A
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- amidoalcohol
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
- A61K8/42—Amides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/10—Washing or bathing preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
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Abstract
The invention discloses natural components for treating melanin pigmentation and application of the natural components in preparing freckle-removing and whitening cosmetics. The research shows that the aurous amidoalcohol ester and the aurous amidoalcohol have the activity of inhibiting the expression of melanin synthesis related protein, can inhibit the activity of tyrosinase, further inhibit the deposition of melanin in cells, and have the effects of removing freckles and whitening, so the aurous amidoalcohol ester and the aurous amidoalcohol have the prospect of being developed into freckles removing and whitening cosmetics, such as freckles removing and whitening facial cleanser, cream or facial mask.
Description
Technical Field
The invention belongs to the field of cosmetics, relates to freckle removing and whitening application of natural ingredients, and particularly relates to natural ingredients for treating melanin pigmentation and application of the natural ingredients in preparing freckle removing and whitening cosmetics.
Background
Melanin is a biological pigment of protein derivatives, which is synthesized by cells themselves and is insoluble in water and soluble in alkaline solution. Melanin can absorb ultraviolet rays and protect skin from skin damage caused by excessive irradiation of ultraviolet rays. However, excessive synthesis of melanin can lead to darkening of skin tone and even the development of cosmetic problems such as freckles.
There are many enzymes involved in melanin synthesis, the most critical one of which is tyrosinase. Under the action of tyrosinase, melanocytes continuously transport melanin granules to cutin. After the human body is irradiated by sunlight ultraviolet rays, tyrosinase is stimulated and activated by the ultraviolet rays, the process of synthesizing melanin by melanocytes is accelerated, and the skin becomes dark.
At present, many scientific researchers still focus on molecular mechanism research in the aspect of melanin synthesis and screening and research and development work of whitening drugs so as to solve the problem of pigmentation of various skins and realize the effects of removing freckles and whitening.
The Chinese violet is a perennial herb, and researches prove that the Chinese violet extract has the effect of resisting skin aging. Golden amidoalcohol ester, golden amidoalcohol and trichosanthes esterine are three natural chemical components existing in Chinese violet, and no research on removing melanin and removing freckle and whitening exists at present.
Disclosure of Invention
The invention aims to provide natural components for treating melanin pigmentation and application of the natural components in preparing freckle-removing and whitening cosmetics.
The above purpose of the invention is realized by the following technical scheme:
a natural component for treating melanin pigmentation comprises aureoyl amide ester or aureoyl amide alcohol.
An application of golden amidoalcohol ester or golden amidoalcohol in removing speckle and whitening skin is disclosed.
An application of golden amidoalcohol ester or golden amidoalcohol in preparing the cosmetics for removing spots and whitening skin is disclosed.
Further, the cosmetic may be a facial cleanser.
Further, the cosmetic may be a cream.
Further, the cosmetic may be a mask.
Has the advantages that:
the invention discovers that the aurous amidoalcohol ester and the aurous amidoalcohol have the activity of inhibiting the expression of melanin synthesis related protein, can inhibit the activity of tyrosinase, further inhibit the deposition of melanin in cells, and have the effects of removing freckles and whitening, so the aurous amidoalcohol ester and the aurous amidoalcohol have the prospect of being developed into freckles removing and whitening cosmetics, such as freckles removing and whitening facial cleanser, cream or facial mask.
Drawings
FIG. 1 shows the melanin content in the melanoma cells of each group; compared with a control group, the contents of melanin in melanoma cells of the golden amidoalcohol ester, golden amidoalcohol and trichosanthes esterine medicine groups are all obviously reduced;
FIG. 2 shows the expression levels of melanin synthesis-related proteins in groups of melanoma cells; compared with a control group, the expression levels of melanin synthesis related proteins TYR and MITF in melanoma cells of the aureoamide alcohol ester, aureoamide alcohol and trichosanthene esterine drug groups are remarkably reduced;
FIG. 3 is a graph showing tyrosinase activity in various groups of melanoma cells; compared with a control group, the tyrosinase activities in melanoma cells of the golden amidoalcohol ester, golden amidoalcohol and trichosanthene esterine drug groups are all obviously reduced.
Detailed Description
The following examples are given to illustrate the essence of the present invention, but not to limit the scope of the present invention.
First, experimental reagent
Mouse melanoma B16-F10 cells were cryopreserved in a laboratory and used after resuscitation.
Fetal bovine serum FBS and DMEM media were purchased from Gibco, usa.
The purities of the test compounds of aureoyl amide ester, aureoyl amide alcohol and trichosanthes kirilowii maxim alkali are more than or equal to 98.5 percent.
The various chemical reagents are conventional.
Second, Experimental methods
1. Cell culture
Mouse melanoma B16-F10 cells were cultured in DMEM complete medium containing 10% fetal bovine serum and 1% double antibody at 37 deg.C with 5% CO2Culturing in a constant temperature incubator, digesting and passing after the cells grow over the bottom of the culture flask.
2. Effect on the Melanin content in melanoma cells
2.1 selection of action concentration
The MTT method is adopted to screen out the concentration which has no obvious proliferation inhibition effect on mouse melanoma B16-F10 cells, and the method comprises the following steps:
logarithmic growth phaseThe mouse melanoma B16-F10 cells are digested and then prepared into cell suspension by using a DMEM complete medium containing 10% fetal calf serum and 1% double antibody, and the cell concentration is 2 × 105Perml, inoculated in a 96-well plate at 100. mu.L per well, 5% CO at 37 ℃2Culturing in a constant temperature incubator. After adaptive culture for 12h, the drug group is replaced by DMEM complete medium containing aureoamide alcohol ester, aureoamide alcohol or trichosanthene base with different concentrations for culture, and cells cultured without adding drugs are set as a control group and a culture solution without cells is set as a blank group. And after continuing culturing for 48h, adding 20 mu L of MTT solution with the concentration of 5mg/mL into each well, incubating for 4h, removing the supernatant, adding 150 mu L of LDMSO into each well, oscillating to fully dissolve crystals, measuring the OD value of each well at the wavelength of 490nm, and calculating the proliferation inhibition rate of different drugs on cells according to a formula. 3 replicates per concentration.
Proliferation inhibition (%) was [ 1- (OD drug-OD blank)/(OD control-OD blank) ] × 100%
The result shows that 25nM golden amidoalcohol ester, golden amidoalcohol or trichosanthes ester base has no obvious proliferation inhibition effect on mouse melanoma B16-F10 cells, and the inhibition rates are respectively (3.16 +/-0.19)%, (3.65 +/-0.22)%, and (3.52 +/-0.17)%, and are all less than 5%. Therefore, a final concentration of 25nM was chosen for subsequent experiments.
2.2 Effect on melanin content
The influence of the drug test on the melanin content in the mouse melanoma B16-F10 cells, which has no obvious proliferation inhibition concentration on the mouse melanoma B16-F10 cells, is tested by the following steps:
taking mouse melanoma B16-F10 cells in logarithmic growth phase, digesting, and preparing a cell suspension by using a DMEM complete culture medium containing 10% fetal calf serum and 1% double antibody, wherein the cell concentration is 2 × 105Perml, inoculated in 6-well plates, 2mL per well, 5% CO at 37 ℃2Culturing in a constant temperature incubator. After adaptive culture for 12h, the drug group was replaced with DMEM complete medium containing 25nM aureoamid alcohol, aureoamid alcohol or trichosanthene base, while the non-drug-cultured cells were set as the control group. After further culturing for 48h, the culture medium is discarded, the cells are digested by PBS, and the cells are added with the culture medium containing 1 mol/ml/or the cells are/isAnd (3) uniformly blowing and beating the L NaOH, carrying out water bath at 80 ℃ for 30min, adding deionized water to enable the concentration of the NaOH to be 0.2mol/L, uniformly blowing and beating, transferring into a 96-well plate, measuring the OD value of each hole at the wavelength of 475nm, and calculating the melanin content of the drug group according to a formula, wherein the melanin content (%) -OD drug group/OD control group is × 100%, and each concentration is 3 multiple holes.
3. Effect on expression of protein involved in melanin synthesis in melanoma cells
Taking mouse melanoma B16-F10 cells in logarithmic growth phase, digesting, and preparing a cell suspension by using a DMEM complete culture medium containing 10% fetal calf serum and 1% double antibody, wherein the cell concentration is 2 × 105Perml, inoculated in 6-well plates, 2mL per well, 5% CO at 37 ℃2After 12h of adaptive culture in a constant temperature incubator, the drug group is replaced by DMEM complete culture medium containing 25nM aureoamid alcohol ester, aureoamid alcohol or trichosanthes kirilowii maxim alkali for culture, meanwhile, cells cultured without adding drugs are set as a control group, after 48h of continuous culture, the culture solution is discarded, PBS is washed, the cells are digested and are cracked on ice, a BCA kit is used for measuring the protein concentration, 50 mu g of protein is taken for SDS-PAGE electrophoresis of each group, the protein is transferred to a PVDF membrane, 5% skimmed milk powder is sealed, MITF, TYR and β -actin primary antibody are added for overnight incubation at 4 ℃, TBS-T washing solution is used for washing the membrane, secondary antibody is added for incubation at room temperature for 1h, ECL method color development and exposure are carried out, and a chemiluminescence imaging.
4. Effect on tyrosinase Activity in melanoma cells
The effect of the drug test on the tyrosinase activity in the mouse melanoma B16-F10 cells, which has no obvious proliferation inhibition concentration on the mouse melanoma B16-F10 cells, is as follows:
taking mouse melanoma B16-F10 cells in logarithmic growth phase, digesting, and preparing a cell suspension by using a DMEM complete culture medium containing 10% fetal calf serum and 1% double antibody, wherein the cell concentration is 2 × 105Perml, inoculated in a 96-well plate at 100. mu.L per well, 5% CO at 37 ℃2Culturing in a constant temperature incubator. After adaptive culture for 12h, the drug group is replaced by DMEM complete medium containing 25nM aureoamidol ester, aureoamidol or trichosanthes kirilowii maxim base, and no drug is addedAfter culturing for 48 hours, the cultured cells were discarded, the culture medium was washed with PBS, 100. mu.L of 0.1% Triton-X-100 (prepared with PBS having a pH of 6.8) solution was added to each well, the cells were completely lysed by shaking for 5min, 50. mu.L of 0.1% L-DOPA (prepared with PBS having a pH of 6.8) solution was added to each well, the reaction was carried out at 37 ℃ for 20min, the OD at a wavelength of 475nm was measured in each well, and based on 100% tyrosinase activity of the control, the drug group tyrosinase activity (OD) was calculated as OD drug group/OD control group × 100% and 3 replicate wells per concentration according to the formula.
5. Data processing
Data were processed using SPSS17.0 statistical software and experimental results are presented as mean ± SD. Group comparisons using the t-test, p <0.05 indicated significant differences.
Third, experimental results
1. Effect on the Melanin content in melanoma cells
As shown in Table 1 and FIG. 1, the content of melanin in melanoma cells was significantly reduced in the golden amidoalcohol, and trichosanthene base drug groups compared to the control group, and the results were statistically different.
TABLE 1 melanin content in groups of melanoma cells
| Group of | Content of melanin |
| Control group | 100％ |
| Golden amide alcohol ester group | (43.6±2.1)％ |
| Golden amide alcohol group | (40.5±2.3)％ |
| Trichosanthes kirilowii maxim ester base group | (52.8±2.7)％ |
2. Effect on expression of protein involved in melanin synthesis in melanoma cells
As shown in FIG. 2, the expression levels of melanin synthesis-related proteins TYR and MITF were significantly decreased in melanoma cells of the auroamide ester, auroamide alcohol, trichosanthene base drug groups, as compared to the control group. It is known to those skilled in the art that the expression and activity of the TYR protein in cells directly determines the melanin content, whereas the TYR protein is regulated by the key transcription factor MITF, both of which are two important related proteins for the synthesis of melanin in melanoma cells.
3. Effect on tyrosinase Activity in melanoma cells
As shown in Table 2 and FIG. 3, the tyrosinase activities in melanoma cells were significantly decreased in the golden amidoalcohol, and trichosanthene base drug groups compared to the control group, and were statistically different.
TABLE 2 tyrosinase activity in groups of melanoma cells
The results show that the aurora amide alcohol ester, the aurora amide alcohol and the trichosanthes ester alkali have the activity of inhibiting the expression of melanin synthesis related protein, can inhibit the activity of tyrosinase, further inhibit the deposition of melanin in cells, and have the effects of removing freckles and whitening, so that the aurora amide alcohol ester, the aurora amide alcohol and the trichosanthes ester alkali have the prospect of being developed into a freckles removing and whitening cosmetic, such as a freckle removing and whitening facial cleanser, a facial cream or a facial mask.
The above-described embodiments are intended to be illustrative of the nature of the invention, but those skilled in the art will recognize that the scope of the invention is not limited to the specific embodiments.
Claims (6)
1. A natural ingredient for treating melanin pigmentation, comprising: is golden amidoalcohol ester or golden amidoalcohol.
2. An application of golden amidoalcohol ester or golden amidoalcohol in removing speckle and whitening skin is disclosed.
3. An application of golden amidoalcohol ester or golden amidoalcohol in preparing the cosmetics for removing spots and whitening skin is disclosed.
4. Use according to claim 3, characterized in that: the cosmetic may be facial cleanser.
5. Use according to claim 3, characterized in that: the cosmetic may be a cream.
6. Use according to claim 3, characterized in that: the cosmetic may be a facial mask.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010479506.0A CN111514048A (en) | 2020-05-30 | 2020-05-30 | Natural component for treating melanin pigmentation and application of natural component in preparing freckle-removing and whitening cosmetics |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202010479506.0A CN111514048A (en) | 2020-05-30 | 2020-05-30 | Natural component for treating melanin pigmentation and application of natural component in preparing freckle-removing and whitening cosmetics |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113913315A (en) * | 2021-12-01 | 2022-01-11 | 广州暨创生物医药研究院有限公司 | Yeast with whitening and freckle removing effects and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5804206A (en) * | 1997-03-06 | 1998-09-08 | Bio-Botanica, Inc. | Therapeutic composition and method for treating skin using Centipeda cunninghami extract |
| CN103479803A (en) * | 2013-09-29 | 2014-01-01 | 陈庆军 | Composition with skin-whitening, acne-removing and anti-allergic effects |
-
2020
- 2020-05-30 CN CN202010479506.0A patent/CN111514048A/en not_active Withdrawn
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5804206A (en) * | 1997-03-06 | 1998-09-08 | Bio-Botanica, Inc. | Therapeutic composition and method for treating skin using Centipeda cunninghami extract |
| CN103479803A (en) * | 2013-09-29 | 2014-01-01 | 陈庆军 | Composition with skin-whitening, acne-removing and anti-allergic effects |
Non-Patent Citations (2)
| Title |
|---|
| PEDRO FONG等: "In Silico Prediction of Tyrosinase and Adenylyl Cyclase Inhibitors from Natural Compounds", 《NATURAL PRODUCT COMMUNICATIONS》 * |
| 徐金钟等: "紫花地丁化学成分研究", 《中草药》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113913315A (en) * | 2021-12-01 | 2022-01-11 | 广州暨创生物医药研究院有限公司 | Yeast with whitening and freckle removing effects and application thereof |
| CN113913315B (en) * | 2021-12-01 | 2024-01-30 | 广州暨创生物医药研究院有限公司 | Yeast with whitening and freckle removing effects and application thereof |
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