Mitochondrial energy improving composition based on plant stem cells, application and cosmetics

Abstract

The invention belongs to the field of daily chemicals, and discloses a mitochondrial energy improving composition based on plant stem cells, application and cosmetics, the composition comprises the following components by weight: 0.0005-1 part of sunflower bud extract; 0.0005 to 1 part of a melissa officinalis extract; and 0.0005 to 1 part of epigallocatechin gallo glucoside or epigallocatechin gallate. According to the composition disclosed by the invention, the sunflower bud extract, the melissa officinalis extract, the epigallocatechin gallyl glucoside or the epigallocatechin gallate are combined, and the combination of the sunflower bud extract, the melissa officinalis extract, the epigallocatechin gallyl glucoside and the epigallocatechin gallate has the effects of obviously improving the cell mitochondrial dysfunction, improving the mitochondrial energy and increasing the content of I-type collagen; the composition can be used as a potential optional scheme of anti-wrinkle active ingredients in cosmetics.

Description

Technical Field

[0001] The invention relates to the field of daily chemicals, and in particular to a mitochondrial energy improving composition based on plant stem cells, its application and cosmetics. Background Art

[0002] Cellular energy ATP has been a hot topic in recent years. As an important energy source for cells, the health of mitochondria directly affects the vitality and lifespan of cells. Therefore, protecting mitochondria and maintaining cell energy have become new focuses in the anti-aging field. It is foreseeable that more anti-aging skin care products and raw material products based on mitochondria will emerge in the market in the future.

[0003] The anti-aging research of lemon balm extract can be found in the following literature: The application number is CN118453476A, and the subject is a patent application for a repair and anti-aging composition and a repair and anti-aging cosmetic; The application number is CN119258200A, and the subject is a patent application for an anti-aging composition for increasing dermal collagen and a preparation method thereof; The anti-aging research on epigallocatechin galloside or epigallocatechin gallate can be found in the following literature: The application number is CN115887246A, and the subject is a patent application for a novel microcapsule composition with multiple anti-aging effects and its preparation method and application; The applicant believes that mitochondrial dysfunction may cause increased oxidative stress and produce excessive reactive oxygen free radicals (ROS). ROS will further damage various components in the cell, including the mitochondria themselves, forming a vicious cycle, which will greatly reduce the energy produced by the mitochondria. Insufficient energy supply will lead to a decrease in the cell metabolic rate, affecting the normal physiological function of the cell, and then affecting the function of tissues and organs, and ultimately accelerating skin aging. Therefore, by removing ROS, mitochondria can be prevented from being damaged by oxidation, mitochondrial dysfunction can be improved, and mitochondrial energy levels can be increased, thereby improving skin aging. The antioxidant (DPPH free radical scavenging) ability of the above substances was verified to evaluate their ability to scavenge free radicals and improve mitochondrial dysfunction.

[0004] During the verification process, we found that the above substances have certain free radical scavenging capabilities, but their ability to improve mitochondrial dysfunction needs to be further improved. Summary of the invention

[0005] The purpose of the present invention is to provide a mitochondrial energy improving composition based on plant stem cells. The composition is a combination of sunflower sprout extract, lemon balm extract, and EGCG (epigallocatechin galloside or epigallocatechin gallate). Compared with the free radical scavenging ability of the three when used alone, the free radical scavenging ability of the composition can be effectively improved; further verification found that the combination of the three has obvious improvement of mitochondrial dysfunction, the ability to increase mitochondrial energy and the content of type I collagen, and can be used as a potential optional solution for anti-wrinkle active ingredients in cosmetics.

[0006] Meanwhile, the invention also provides applications and cosmetics based on the composition.

[0007] The specific scheme of the present invention is: A mitochondrial energy improving composition based on plant stem cells, comprising the following components in parts by weight: Sunflower sprout extract 0.0005~1 part; 0.0005~1 part of lemon balm extract; Epigallocatechin galloside or epigallocatechin gallate 0.0005-1 part.

[0008] In some embodiments of the present invention, the amount of the sunflower sprout extract can be selected to be 0.0005 parts, 0.001 parts, 0.005 parts, 0.01 parts, 0.05 parts, 0.1 parts, 0.5 parts, 1 parts; The amount of the lemon balm extract can be selected to be 0.0005 parts, 0.001 parts, 0.005 parts, 0.01 parts, 0.05 parts, 0.1 parts, 0.5 parts, 1 parts; The amount of epigallocatechin galloside or epigallocatechin gallate can be selected to be 0.0005 part, 0.001 part, 0.005 part, 0.01 part, 0.05 part, 0.1 part, 0.5 part, 1 part; In the present invention, the lemon balm extract is a plant stem cell extract. When the lemon balm extract is used alone, its free radical scavenging rate is very low; when EGCG is used alone, its free radical scavenging effect is more obvious than that of any one of the sunflower sprout extract and the lemon balm extract; when EGCG is matched with any one of the sunflower sprout extract and the lemon balm extract, a certain synergy can be exhibited, among which the synergy between EGCG and the lemon balm extract is more obvious; In subsequent experiments of the present invention, it was found that in the presence of EGCG, the synergistic effect of sunflower sprout extract and lemon balm extract when used in combination can be further improved.

[0009] When performing mitochondrial membrane potential testing, we further demonstrated a potential direct correlation between better free radical scavenging and mitochondrial membrane potential.

[0010] We believe that the underlying mechanism is: Mitochondrial dysfunction usually refers to energy metabolism disorders caused by factors such as mitochondrial membrane damage, respiratory chain obstruction, decreased enzyme activity, and mitochondrial DNA (mtDNA) damage, which can trigger a series of chain reactions and cause cell damage. Further research revealed that oxidative stress is a key factor leading to mitochondrial dysfunction.

[0011] Sunflower sprout extract improves the function of the mitochondrial respiratory chain by increasing NAD+ levels, while lemon balm extract maintains the morphological integrity of mitochondria by protecting them from damage by reactive oxygen species (ROS). EGCG, as the core active ingredient in green tea, is known as the "diamond molecule in green tea". It has extremely strong antioxidant capacity, which is 25 times that of vitamin C and 3 times that of pomegranate seed extract. The strong antioxidant effect of EGCG can further protect mitochondria from oxidative damage by free radicals. It can be inferred from the results that under the action of EGCG, the health of mitochondria will be significantly improved, showing a significant synergistic effect. From the characteristics of each raw material, it can be inferred that the mechanism of improvement of mitochondrial energy of the composition is based on the powerful antioxidant activity of EGCG to protect mitochondria from oxidative damage by free radicals, the cellular complex contained in the lemon balm extract preserves the integrity of cellular nutrients and its antioxidant capacity to further ensure the health of mitochondria, and the sunflower sprout extract strengthens the skin's output of NAD+ to increase ATP levels, thereby improving DNA repair function, optimizing cell function, and increasing cell vitality; through the above three active ingredients resisting external damage, maintaining the integrity of the cells themselves, and repairing DNA inside the cells, the multiple coordination and synergy from the outside to the inside achieves the improvement of mitochondrial energy, thereby improving skin wrinkles.

[0012] At the same time, tests on the expression of type I collagen of each substance found that sunflower sprout extract, lemon balm extract, and EGCG can all show a certain effect of enhancing the expression of type I collagen, among which the effect of lemon balm extract is more obvious. When the three are used in combination, the effect is significantly improved.

[0013] The above composition also includes one or more combinations of alpine edelweiss callus extract, polyphylla root extract, and saponaria extract.

[0014] Alpine edelweiss callus extract, syngonium root extract, and saponaria extract are also plant stem cell extracts. They have been used in a moisturizing product of the applicant and have achieved good results. In the research process of this application, it is hoped that they can show certain effects in the formula of the present invention. Based on the experimental results that have been conducted, their free radical scavenging ability is limited and has no negative impact on mitochondrial membrane potential. Through the expression test experiment of type I collagen, it can be seen that alpine edelweiss callus extract, syngonium root extract, and saponaria extract can improve the expression of type I collagen to a certain extent. When used in combination with sunflower sprout extract, lemon balm extract, and EGCG, they can play a more obvious role in improving the expression of type I collagen. This also shows that skin anti-wrinkle is the result of the combined action of multi-faceted and multi-channel factors. Although the effects of alpine edelweiss callus extract, polychaete root extract, soapwort extract, sunflower sprout extract, lemon balm extract, and EGCG on mitochondrial membrane potential are superimposed and have no synergistic effect, they have obvious synergy in the expression of type I collagen.

[0015] In the above composition, the amount of the alpine edelweiss callus extract is 0 to 1 part by weight; The amount of the comfrey root extract is 0 to 1 part by weight; The amount of the soapwort extract is 0 to 1 part by weight.

[0016] In the above composition, the amount of the alpine edelweiss callus extract is 0.001 to 1 part by weight; The amount of the comfrey root extract is 0.001 to 1 part by weight; The dosage of the soapwort extract is 0.001 to 1 part by weight.

[0017] At the same time, the present invention also discloses the use of any of the above compositions in preparing cosmetics.

[0018] In the above-mentioned use, the cosmetic is a cosmetic used to enhance the energy of cell mitochondria.

[0019] In addition, the present invention also discloses a cosmetic comprising any one of the above compositions.

[0020] The above-mentioned cosmetic contains 0.001-10 wt % of the composition.

[0021] In the above-mentioned cosmetics, the dosage forms of the cosmetics at least include lotion, essence, lotion, emulsion, cream and mask.

[0022] The beneficial effects of this application are: The composition of the present invention adopts a combination of sunflower sprout extract, lemon balm extract, epigallocatechin galloside or epigallocatechin gallate to effectively improve the free radical scavenging ability of the composition; further verification found that the combination of the three has the ability to significantly improve mitochondrial dysfunction and increase mitochondrial energy, thereby improving the skin's anti-wrinkle ability. DETAILED DESCRIPTION

[0023] The present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. In the description of the present invention, it should be noted that, if no specific conditions are specified in the embodiments, the conditions are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0024] Part I Reagent source description: The sunflower sprout extract was purchased from Guangzhou Lingyu Trading Co., Ltd., with a content of 0.1-5wt%; The lemon balm extract was purchased from Guangzhou Aoxue Chemical Co., Ltd., with a content of 3-10wt%; The EGCG was purchased from Guangzhou Aoxue Chemical Co., Ltd., with a content of 5-10wt%; The alpine edelweiss callus extract was purchased from Guangzhou Aoyuan Trading Co., Ltd., with a content of 15-30wt%; The polysaccharide root extract was purchased from Guangzhou Lingyu Trading Co., Ltd., with a content of 0.5-1%wt; The soapwort extract was purchased from Guangzhou Lingyu Trading Co., Ltd., with a content of 0.5-1%wt; Actual purchases may occur from different manufacturers and the content may fluctuate slightly.

[0025] Part 2

[0026] Example 1

[0027] The preparation method of the composition of the present invention is: Dissolve sunflower sprout extract, lemon balm extract, EGCG, alpine edelweiss callus extract, polysaccharide root extract, and soapwort extract in water and stir until completely dissolved.

[0028] The formula of the composition is shown in Table 1; Table 1 Formula Unit: g Sunflower sprout extract Melissa officinalis Extract EGCG Leontopodium alpinum Callus Extract Polygonum aviculare root extract Soapwort Extract water Example 1 0.0005 0.0005 0.0005 0 0 0 To 100 Example 2 1 1 1 0 0 0 To 100 Example 3 0.0005 0.01 0.5 0 0 0 To 100 Example 4 0.01 0.0005 0.8 0 0 0 To 100 Example 5 0.5 0.5 0.0005 0 0 0 To 100 Example 6 0.01 0.01 0.002 0 0 0 To 100 Example 7 0.01 0.01 0.002 0.001 0.001 0.001 To 100 Example 8 0.01 0.01 0.002 1 1 1 To 100 Example 9 0.01 0.01 0.002 0.5 0.5 0.5 To 100 Comparative Example 1 0 0 0 0.001 0.001 0.001 To 100 Comparative Example 2 0 0 0 0.5 0.5 0.5 To 100 Comparative Example 3 0.01 0.01 0 0 0 0 To 100 Comparative Example 4 0.02 0 0.002 0 0 0 To 100 Comparative Example 5 0 0.02 0.002 0 0 0 To 100 Comparative Example 6 0.02 0 0 0 0 0 To 100 Comparative Example 7 0 0.02 0 0 0 0 To 100 Comparative Example 8 0 0 0.002 0 0 0 To 100

[0029] Part 3 3.1 DPPH free radical scavenging rate Weigh 0.005g of DPPH (1,1-diphenyl-2-picrylhydrazyl) powder, add an appropriate amount of anhydrous ethanol to dissolve, and dissolve it completely by ultrasound in the dark. Then dilute it to 100mL with anhydrous ethanol to prepare a 50.0µg / mL DPPH solution. Use purified water to prepare each example and comparative example sample, 1mL of sample in the experimental group is mixed with 3mL of DPPH solution, 1mL of sample in the control group is mixed with 3mL of anhydrous ethanol, and 1mL of sample solvent in the blank group is mixed with 3mL of DPPH solution. After fully mixing, incubate in the dark for 30min. Add 200μL of the reaction solution of each group to a 96-well plate, with 3 parallel wells in each tube, and measure the absorbance at 517nm.

[0030] The free radical scavenging rate was calculated according to the following formula.

[0031] Clearance rate (%) = [1-(A 实验组 -A 对照组 ) / A 空白组 ]×100%; A 实验组 ——The absorbance of the mixture of the sample solution and DPPH solution; A 对照组 ——The absorbance of the mixture of the sample solution and the anhydrous ethanol solution; A 空白组 —— The absorbance of the mixture of DPPH solution and sample solvent solution.

[0032] The DPPH free radical scavenging rate reflects its antioxidant capacity to a certain extent. The higher the scavenging rate, the better the antioxidant degree and the better the anti-wrinkle effect. Therefore, the anti-wrinkle effect of the composition can be judged by studying its ability to scavenge DPPH free radicals.

[0033] 3.2 Mitochondrial membrane potential HaCaT cells were seeded in 12-well plates (5 × 10 4 The cells were incubated at 37 °C, 5% CO2 for 24 h. The normal control group, model control group, and sample group were set up in the experiment. The culture medium was removed, and the cells were washed with D-Hanks 1-2 times. The sample group and the model control group were subjected to UVB modeling (100 mJ / cm 2 ). After the modeling was completed, D-Hanks was washed 1-2 times, and complete culture medium (the final content of each component is shown in Table 1) was added to the normal control group and the model control group, and complete culture medium containing corresponding concentrations of the examples and comparative examples was added to the sample group, and cultured for 24 h at 37°C and 5% CO2. After the culture was completed, JC-1 staining was performed and the cells were observed and photographed under a fluorescence microscope. Image J was used to analyze the fluorescence intensity (S) of each group, and the mitochondrial membrane potential was expressed by the red / green fluorescence intensity ratio. The mitochondrial membrane potential (red / green fluorescence intensity ratio) was calculated.

[0034] Mitochondrial membrane potential (ratio of red / green fluorescence intensity) = S (red fluorescence intensity) / S (green fluorescence intensity).

[0035] The samples used for mitochondrial membrane potential are Example 6, Example 7 and Comparative Example 1.

[0036] The mitochondrial membrane potential increase rate was calculated according to the following formula: Mitochondrial membrane potential enhancement rate = (mitochondrial membrane potential of sample group - mitochondrial membrane potential of model control group) / mitochondrial membrane potential of model control group × 100% 3.3 Type I collagen content ESF-M cells were inoculated in a 24-well plate and incubated at 37°C, 5% CO2 for 24 h. The experiment set up a normal control group, a model control group and a sample group. The culture medium in the well was discarded, and the sample group was added with complete culture medium containing corresponding concentrations of the examples and comparative examples (the final content of each component is shown in Table 1), and continued to be cultured at 37°C, 5% CO2 for 24 h; the cell plate was removed, the supernatant was discarded, and the cells were washed with PBS, and the cells were exposed to UVA for irradiation; after the irradiation, the PBS was discarded, and each group was added with complete culture medium and placed in a cell culture incubator for continued culture. Remove the culture plate, discard the culture medium in the well, add PBS to wash and discard; Add paraformaldehyde fixative to fix the cells, add PBS to wash and discard; Add Triton X-100 solution to permeabilize, add PBS to wash and discard; Add bovine serum albumin solution to block, add PBS to wash and discard; Add Rabbit Anti-Collagen I antibody and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) for staining and incubation, add PBS to wash and discard; DAPI staining solution, add PBS to wash and discard, add appropriate amount of anti-fluorescence quencher; Observe and take pictures under a fluorescence microscope. Use ImageJ to count the fluorescence intensity of each group of green fluorescence images.

[0037] The increase rate of type I collagen content = (green fluorescence intensity of the sample group - green fluorescence intensity of the model control group) / green fluorescence intensity of the model control group × 100%.

[0038] The test results can be seen in Table 2; Table 2 Test result statistics DPPH free radical scavenging rate% Mitochondrial membrane potential increase rate% Increase rate of type I collagen content% Example 1 20.50 / / Example 2 91.38 / / Example 3 95.84 / / Example 4 95.79 / / Example 5 83.96 / / Example 6 50.43 532 235.14 Example 7 62.75 571 386.75 Example 8 81.85 / / Example 9 75.87 / / Comparative Example 1 5.36 98 81.65 Comparative Example 2 23.31 / / Comparative Example 3 6.77 / / Comparative Example 4 30.11 / / Comparative Example 5 40.15 / / Comparative Example 6 4.01 / 76.52 Comparative Example 7 7.82 / 57.88 Comparative Example 8 20.67 / 96.04

[0039] Result analysis: 1. According to the test results of Example 1, Example 2, Example 3, Example 4, Example 5, and Example 6, the free radical scavenging ability is closely related to the concentration of the effective ingredient; from the experimental results of Example 3, Example 4, and Example 5, when the dosage of any one of the sunflower sprout extract, the lemon balm extract, and the EGCG is very small, the composition can show a relatively excellent free radical scavenging effect; among them, the effect of Example 5 is relatively weak, the reason is that among the three, EGCG has the strongest single antioxidant ability, and when its dosage is small, the dosage of the other two is required to increase in order to achieve a relatively good free radical scavenging effect; this conclusion can be supported by the data of the free radical scavenging ability of Comparative Example 8.

[0040] 2. The test results of Example 6 and Comparative Examples 3 to 5 prove that EGCG is the main free radical scavenging component in the form of two combinations; 3. Combining the data of Comparative Examples 6 to 8 and Comparative Examples 3 to 5, there is no obvious synergy between the sunflower sprout extract, the lemon balm extract, and EGCG; from the results of Example 6 and Comparative Examples 6 to 8, there is obvious internal synergy between the above three; Judging from the test results of Comparative Examples 3 and 8, Comparative Examples 4 and 7, and Comparative Examples 5 and 6, sunflower sprout extract is the core of synergy. From the preliminary research of this application, it can be concluded that sunflower sprout extract has the weakest free radical scavenging ability among all substances, and its ability to significantly enhance the antioxidant properties of the latter two exceeds the expectations of the preliminary experimental design.

[0041] 4. Combining the results of Comparative Examples 6, 7, 1, 9 and 2, the free radical scavenging ability of alpine edelweiss callus extract, polysaccharide root extract and saponaria extract requires a certain concentration. This result can be concluded from the data change trend of Examples 7 to 9 and Comparative Examples 1 to 2. It can be inferred that the mitochondrial membrane potential enhancement rate may show the same trend. Based on the performance evaluation results of the applicant's products already on sale, even if it does not show a good free radical scavenging effect at a low concentration, it will effectively compensate for the composition of the present invention in terms of moisturizing and other properties.

[0042] 5. According to the test results of the increase rate of type I collagen content, compared with the use of a single ingredient among sunflower sprout extract, lemon balm extract, and EGCG, the expression of type I collagen in Example 6 has been significantly improved. At the same time, referring to the results of Example 6 and Example 7 and Comparative Example 1, the alpine edelweiss callus extract, the polysaccharide root extract, and the soapwort extract can increase the expression of type I collagen to a certain extent. When used in combination with sunflower sprout extract, lemon balm extract, and EGCG, they can play a more obvious role in increasing the content of type I collagen.

[0043] 3.3 Security Testing Select qualified patch test materials. Put the test substance (Example 2, Example 8) into the patch tester, with a dosage of about 0.020mL-0.025mL (the liquid is dripped on the filter paper attached to the patch tester and placed in the patch tester). Apply the patch tester with the test substance to the back or forearm of the subject with a non-irritating tape, and use the palm of the hand to gently press it to evenly apply it to the skin for 24 hours.

[0044] The criteria are shown in Table 3 (Evaluation criteria for skin reactions to closed patch tests); Table 3 Evaluation criteria for skin reactions to closed patch test Degree of reaction Rating Skin reactions - 0 Negative reaction ± 1 Suspected reaction: only slight erythema + 2 Weak positive reaction (erythema reaction): erythema, infiltration, edema, and papules may be present ++ 3 Strong positive reaction (herpes reaction): erythema, infiltration, edema, papules, herpes; the reaction may extend beyond the test area +++ 4 Very strong positive reaction (confluent herpes reaction): obvious erythema, severe infiltration, edema, confluent herpes; reaction beyond the test area

[0045] Result determination: When more than 5 out of 30 subjects had grade 1 adverse skin reactions, or more than 2 had grade 2 adverse skin reactions (more than 5 subjects had grade 2 reactions in the deodorant product patch test), or any grade 3 or above adverse skin reaction occurred, the test substance was determined to have adverse skin reactions in humans. The test results can be seen in Table 4.

[0046] Table 4 Test results of closed patch skin reaction test

[0047] Conclusion: The human patch test results showed that: The skin of the test subject in Example 30 showed a negative reaction (scoring level 0). Referring to the human patch test judgment standard of "HAT-SOP-F1-PZ-092", Example 2 and Example 8 had no adverse reaction to human skin.

Claims

1. A mitochondrial energy improving composition based on plant stem cells, characterized in that: It includes the following components by weight: Sunflower sprout extract 0.0005~1 part; 0.0005~1 part of lemon balm extract; Epigallocatechin galloside or epigallocatechin gallate 0.0005-1 part.

2. The composition according to claim 1, characterized in that It also includes one or more combinations of alpine edelweiss callus extract, polycotton root extract, and soapwort extract.

3. The composition according to claim 2, characterized in that The amount of the alpine edelweiss callus extract is 0-1 part by weight; The amount of the comfrey root extract is 0 to 1 part by weight; The amount of the soapwort extract is 0 to 1 part by weight.

4. The composition according to claim 3, characterized in that The amount of the alpine edelweiss callus extract is 0.001 to 1 part by weight; The amount of the comfrey root extract is 0.001 to 1 part by weight; The dosage of the soapwort extract is 0.001 to 1 part by weight.

5. Use of the composition according to any one of claims 1 to 4 in preparing cosmetics.

6. The use according to claim 5, characterized in that The cosmetic is a cosmetic used for improving the energy of cell mitochondria.

7. A cosmetic, characterized in that: Contains the composition according to any one of claims 1 to 4.

8. The cosmetic according to claim 7, characterized in that: A composition containing 0.001 to 10 wt %.

9. The cosmetic according to claim 7, characterized in that: The dosage form of the cosmetic is lotion, essence, lotion, emulsion, cream or facial mask.

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