Composition with skin anti-aging effect and application thereof

Abstract

The invention belongs to the technical field of cosmetics, and particularly relates to a composition with a skin anti-aging effect and application thereof. The composition comprises micromolecular collagen peptide and calendula exosome, and the weight ratio of the micromolecular collagen peptide to the calendula exosome is (1-10): (10-1). In the composition disclosed by the invention, the micromolecular collagen peptide and the calendula exosome in a specific dosage ratio are combined, so that the composition disclosed by the invention has synergistic effects of moisturizing, repairing a skin barrier and resisting wrinkles and tightening, and when the composition is applied to cosmetics, the skin cuticle and barrier function reconstruction can be promoted; the moisturizing property and the water resistance of the skin are improved, the wrinkles of the skin are reduced, the skin is more compact, and the use feeling is excellent.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a composition with anti-aging effects on the skin and its uses. Background Technology

[0002] Aging, prolonged exposure to adverse environmental factors, nutritional imbalances, and fatigue can alter the skin's visual appearance, physical properties, and physiological functions. The most significant and noticeable changes include the formation of fine lines and wrinkles, decreased elasticity, sagging skin, reduced firmness, uneven skin tone or dullness, rough skin texture, and the appearance of age spots. Changes that occur with skin aging or long-term environmental damage include a general decrease in cell and tissue activity, a reduced cell replication rate, decreased blood flow to the skin, decreased moisture content, accumulated structural and functional errors, changes in skin metabolism, and a reduced ability for the skin to rebuild and repair itself. Many changes in the skin's appearance and barrier function are primarily caused by alterations in the outer epidermis, while other changes, such as wrinkles and sagging skin, are mainly caused by changes in the underlying dermis.

[0003] To prevent these phenomena and maintain healthier and more elastic skin, the inventors have been dedicated to researching and developing cosmetics containing various highly active substances to maintain the skin's inherent functions and activate skin cells, thereby effectively slowing down skin aging.

[0004] Small molecule collagen peptides are low-molecular-weight bioactive peptides with special functions, obtained through deep hydrolysis. Compared with proteins, small molecule collagen peptides have a higher absorption rate and stronger bioactivity, while also exhibiting high safety and good biocompatibility. Based on these excellent properties, people increasingly favor using small molecule collagen peptides to delay skin aging. However, in practical applications, it has been found that even with their smaller molecular weight, small molecule collagen peptides still need to penetrate the stratum corneum and epidermis to reach the dermis and exert their effects. The skin's own barrier function hinders the penetration of active substances; therefore, most commercially available collagen peptides currently suffer from low actual absorption rates and less than ideal results.

[0005] In recent years, plant-derived vesicle-like particles (PDEVLPs) have attracted increasing attention due to their potential bioactive effects, becoming a research hotspot in the biomedical and skincare fields. Numerous studies have identified and reported the presence of PDEVLPs in vegetables, fruits, and medicinal plants. PDEVLPs offer advantages such as wide availability, high yield, simple production, and low immune risk, making them a safer and more economical option for development into various functional active substances or carriers.

[0006] In view of the above-mentioned deficiencies in the prior art, the inventors have developed a composition with skin anti-aging effects that simultaneously moisturizes, repairs the skin barrier, and has anti-wrinkle and firming effects. This composition can be conveniently used in various cosmetics to impart the above-mentioned effects to the target cosmetics. Summary of the Invention

[0007] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a composition with skin-anti-aging effects, including moisturizing, skin barrier repair, anti-wrinkle, and firming properties, as well as its preparation method and uses. In this invention's composition, by combining small-molecule collagen peptides and calendula exosomes in specific proportions, the problems of low absorption rate and unsatisfactory effects of small-molecule collagen peptides are effectively solved, resulting in a composition with synergistic moisturizing, skin barrier repair, anti-wrinkle, and firming effects.

[0008] Specifically, the present invention is achieved through the following technical solutions: In a first aspect, the present invention provides a composition having anti-aging effects on the skin, the composition comprising small molecule collagen peptides and calendula exosomes, wherein the weight ratio of the small molecule collagen peptides to the calendula exosomes is 1~10:10~1.

[0009] Alternatively, in the above composition, the weight ratio of the small molecule collagen peptides to calendula exosomes is 1~8:8~1.

[0010] Alternatively, in the above composition, the weight ratio of the small molecule collagen peptides to calendula exosomes is 1~4:4~1.

[0011] Alternatively, in the above composition, the weight ratio of the small molecule collagen peptides to calendula exosomes is 2:3.

[0012] Alternatively, in the above composition, the composition consists of small molecule collagen peptides and calendula exosomes.

[0013] Alternatively, in the above composition, the small molecule collagen peptide is isolated and identified from the enzymatic hydrolysis product of deep-sea cod skin, and the amino acid sequence of the small molecule collagen peptide is HPTAE (His-Pro-Thr-Ala-Glu).

[0014] Alternatively, in the above composition, the small molecule collagen peptides are prepared by solid-phase synthesis.

[0015] Alternatively, in the above composition, the calendula exosomes have a double-membrane structure under a transmission electron microscope, mostly appearing as concave discs, and the particle size of the calendula exosomes is 50-300 nm.

[0016] Alternatively, in the above composition, the calendula exosome primary product can be prepared by differential centrifugation and sedimentation, by ultrafiltration, or by kit extraction, and then purified by at least one of density gradient chromatography, column chromatography, ultrafiltration, etc.

[0017] Alternatively, in the above composition, the method for preparing the calendula exosomes includes the following steps: (1) After washing the fresh calendula flowers with pure water, add an appropriate amount of PBS to soak them. After soaking for a long time, crush them in a blender and filter them through a gauze to obtain filtrate a. (2) Centrifuge filtrate a at 1,000 g for 10-15 min to obtain supernatant a, and centrifuge supernatant a at 3,000 g for 30-40 min to obtain supernatant b; (3) The supernatant b was concentrated by centrifugation at 3,500 g for 30-40 min using a 100 kDa ultrafiltration tube to obtain supernatant concentrate c. Supernatant concentrate c was then centrifuged at 10,000 g for 30-40 min to obtain supernatant concentrate d. (4) The supernatant concentrate d was filtered through a 0.45 μm filter membrane to obtain filtrate b; (5) The filtrate b is centrifuged at 100,000~120,000 g for 60~90 min to obtain a precipitate, which is the calendula exosome.

[0018] Alternatively, in the above composition, the method for preparing the calendula exosomes includes the following steps: (1) After washing the fresh calendula flowers with pure water, add an appropriate amount of PBS to soak them. After soaking for a long time, crush them in a blender and filter them through a gauze to obtain filtrate a. (2) Centrifuge filtrate a at 1,000 g for 12 min to obtain supernatant a, and centrifuge supernatant a at 3,000 g for 30 min to obtain supernatant b; (3) The supernatant b was concentrated by centrifugation at 3,500 g for 30 min using a 100 kDa ultrafiltration tube to obtain supernatant concentrate c. Supernatant concentrate c was then centrifuged at 10,000 g for 30 min to obtain supernatant concentrate d. (4) The supernatant concentrate d was filtered through a 0.45 μm filter membrane to obtain filtrate b; (5) The filtrate b was centrifuged at 120,000 g for 70 min to obtain a precipitate, which is the calendula exosome.

[0019] Alternatively, in the above composition, small molecule collagen peptides and calendula exosomes are resuspended in filtered sterile PBS solution, mixed evenly, and then lyophilized.

[0020] In a second aspect, the present invention provides the use of the composition with anti-aging effects described in the first aspect above in the preparation of cosmetics with moisturizing and skin barrier repairing effects as well as anti-wrinkle and firming effects.

[0021] Alternatively, in the above-described uses, the cosmetic product may be a toner, lotion, serum, cream, ointment, mask, or freeze-dried powder.

[0022] Alternatively, in the above-mentioned uses, the cosmetic has a protective effect against UV-induced cell damage, possesses antioxidant activity, repairs damaged cells, protects the skin barrier, and delays skin aging.

[0023] Alternatively, in the above-described applications, the content of the composition in the cosmetic product is 1%-5% by weight.

[0024] Compared with the prior art, the present invention has the following advantages: (1) By screening the enzymatic hydrolysate of deep-sea cod skin, the inventors obtained for the first time a small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu) that simultaneously protects cell activity, inhibits oxidative damage, protects the skin barrier, enhances collagen synthesis, inhibits collagen degradation, and repairs skin damage, without any obvious side effects.

[0025] (2) In the composition of the present invention, by combining small molecule collagen peptides and calendula exosomes in a specific ratio, the problem of low absorption rate and unsatisfactory effect of small molecule collagen peptides is well solved, so that the composition of the present invention has synergistic effects of moisturizing and repairing the skin barrier as well as anti-wrinkle and firming.

[0026] (3) When the composition of the present invention is applied to cosmetics, it can promote the reconstruction of the skin stratum corneum and barrier function, improve the skin's moisturizing and water-resistant properties, reduce skin wrinkles and make the skin firmer, and has an excellent user experience. Detailed Implementation

[0027] The present invention will be further described below with reference to specific embodiments. It should be understood that the specific embodiments described herein are for illustrative purposes only and are not intended to limit the scope of the invention.

[0028] Where specific techniques or conditions are not specified in the examples, they shall be performed in accordance with the techniques or conditions described in the literature in this field, or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased through legitimate channels.

[0029] Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods. Unless otherwise specified, the experimental materials used in the following embodiments are commercially available products.

[0030] Unless otherwise stated, all percentages and parts in this invention refer to weight percentages and weight parts.

[0031] Preparation Example: 1. Preparation of small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu) 1.1 Preparation of small molecule collagen peptides Take an appropriate amount of frozen deep-sea cod skin, thaw it with water at a ratio of 1:3 (w / v), then soak it in 5% NaOH to remove impurities, rinse it several times with clean water until the pH of the washing water is neutral, add 0.06% alkaline protease and 0.04% bromelain, and enzymatically hydrolyze for 5 h, then inactivate the enzyme in a boiling water bath for 15 min, take the enzymatic hydrolysate and pass it through a 200-mesh silk cloth to remove impurities, filter to remove suspended impurities, then centrifuge at 7500 rpm and 4℃ for 10 min, take the supernatant, freeze dry, and the deep-sea cod skin enzymatic hydrolysate is obtained.

[0032] 1.2 Screening and Identification of Small Molecule Collagen Peptide Sequences The deep-sea cod skin enzymatic hydrolysate obtained in step 1.1 above was desalted and then subjected to chromatographic separation and mass spectrometry detection.

[0033] The specific conditions were as follows: LC-MS / MS analysis was performed using a dual ternary high-performance liquid chromatography-tandem quadrupole orbital trap high-resolution mass spectrometer. The liquid chromatography system employed a reversed-phase system, with 0.1% formic acid in water as the elution phase A and acetonitrile as the elution phase B. The chromatographic column was a C18, 100 × 2.1 mm Waters column, with a flow rate of 0.2 mL / min, a column temperature of 40 ℃, an injection volume of 2 μL, and the mass spectrometry mode set to positive ion. The primary mass spectrometry scan range was 150–1500 m / z, with a resolution of 35,000 and a maximum injection time of 100 ms. The MS2 scan resolution was 17,500, with a maximum injection time of 50 ms, a collision energy of 35 eV, a capillary spray voltage of 3.0 kV, a capillary temperature of 320 ℃, an auxiliary heating temperature of 350 ℃, and an S-lens Rf set to 60. Ten fragment maps were acquired after each full scan (MS2 scan).

[0034] Database retrieval: The raw mass spectrometry test file is retrieved from the corresponding database using the MaxQuant software to obtain the protein identification and quantitative analysis results.

[0035] The peptide sequences of the enzymatic hydrolysate of deep-sea cod skin were identified by mass spectrometry. Peptide sequences with relatively high content and high solubility, non-toxicity, and good stability, as predicted by database retrieval, were selected for further research.

[0036] 1.3 Preparation of HPTAE (His-Pro-Thr-Ala-Glu) by solid-phase synthesis The resin was swollen, washed, and dried to remove the Fmoc protecting group. The amino acids that make up the peptide were weighed and reacted in a shaker at 30°C for 1 hour to carry out a condensation reaction. The process of removal-protection-condensation was repeated until all amino acids were linked. The target peptide was separated from impurities by high-performance liquid chromatography (HPLC) to obtain a pure peptide (>99%), and the target peptide was lyophilized into powder.

[0037] In addition, the same method was used to synthesize the small molecule collagen peptide HPSAE (His-Pro-Ser-Ala-Glu), which serves as a control in the "Effective Examples" section below.

[0038] 2. Preparation of Calendula exosomes The method for preparing the calendula exosomes includes the following steps: (1) After washing the fresh calendula flowers with pure water, add an appropriate amount of PBS to soak them. After soaking for a long time, crush them in a blender and filter them through a gauze to obtain filtrate a. (2) Centrifuge filtrate a at 1,000 g for 12 min to obtain supernatant a, and centrifuge supernatant a at 3,000 g for 30 min to obtain supernatant b; (3) The supernatant b was concentrated by centrifugation at 3,500 g for 30 min using a 100 kDa ultrafiltration tube to obtain supernatant concentrate c. Supernatant concentrate c was then centrifuged at 10,000 g for 30 min to obtain supernatant concentrate d. (4) The supernatant concentrate d was filtered through a 0.45 μm filter membrane to obtain filtrate b; (5) The filtrate b was centrifuged at 120,000 g for 70 min to obtain a precipitate, which is the calendula exosome.

[0039] In addition, Rhodiola rosea exosomes, which serve as a control in the "Effective Examples" section below, were also prepared using the same method.

[0040] 3. Preparation of Example 1 A composition with anti-aging effects on the skin, the composition comprising a small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu) and calendula exosomes, wherein the weight ratio of the small molecule collagen peptide to the calendula exosomes is 2:3.

[0041] Small molecule collagen peptides and calendula exosomes were resuspended in filtered sterile PBS solution, mixed thoroughly, and then lyophilized.

[0042] 4. Preparation of Example 2 A composition with anti-aging effects on the skin, the composition comprising a small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu) and calendula exosomes, wherein the weight ratio of the small molecule collagen peptide to the calendula exosomes is 1:4.

[0043] Small molecule collagen peptides and calendula exosomes were resuspended in filtered sterile PBS solution, mixed thoroughly, and then lyophilized.

[0044] 5. Preparation of Example 3 A composition with anti-aging effects on the skin, the composition comprising a small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu) and calendula exosomes, wherein the weight ratio of the small molecule collagen peptide to the calendula exosomes is 4:1.

[0045] Small molecule collagen peptides and calendula exosomes were resuspended in filtered sterile PBS solution, mixed thoroughly, and then lyophilized.

[0046] 6. Preparation of Comparative Example 1 Only 2 parts by weight of the small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu) were used, resuspended in filtered sterile PBS solution, and then lyophilized.

[0047] 7. Preparation of Comparative Example 2 Using only 3 parts by weight of calendula exosomes, resuspend in filtered sterile PBS solution and lyophilize.

[0048] 8. Preparation of Comparative Example 3 Only 2 parts by weight of the small molecule collagen peptide HPSAE (His-Pro-Ser-Ala-Glu) were used, resuspended in filtered sterile PBS solution, and then lyophilized.

[0049] 9. Preparation of Comparative Example 4 Only 3 parts by weight of Rhodiola rosea exosomes were used, resuspended in filtered sterile PBS solution, and lyophilized.

[0050] 10. Preparation of Comparative Example 5 A composition comprising a small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu) and Rhodiola rosea exosomes, wherein the weight ratio of the small molecule collagen peptide to the Rhodiola rosea exosomes is 2:3.

[0051] Small molecule collagen peptides and Rhodiola rosea exosomes were resuspended in filtered sterile PBS solution, mixed thoroughly, and then lyophilized.

[0052] 11. Preparation of Comparative Example 6 A composition comprising a small molecule collagen peptide HPSAE (His-Pro-Ser-Ala-Glu) and calendula exosomes, wherein the weight ratio of the small molecule collagen peptide to the calendula exosomes is 2:3.

[0053] Small molecule collagen peptides and calendula exosomes were resuspended in filtered sterile PBS solution, mixed thoroughly, and then lyophilized.

[0054] 12. Preparation of Comparative Example 7 A composition comprising a small molecule collagen peptide HPSAE (His-Pro-Ser-Ala-Glu) and Rhodiola rosea exosomes, wherein the weight ratio of the small molecule collagen peptide to the Rhodiola rosea exosomes is 2:3.

[0055] Small molecule collagen peptides and Rhodiola rosea exosomes were resuspended in filtered sterile PBS solution, mixed thoroughly, and then lyophilized.

[0056] Example of effect: Example 1: Effect of the composition of the present invention with anti-aging skin function on the survival rate of UV-damaged HACAT cells. The amount of endogenous adenine triphosphate (ATP) in cells can reflect cell viability. Cell viability was quantitatively detected using ATP. The HACAT cells used in Example 1 were purchased from Wuhan Pronosei Life Sciences Co., Ltd. HACAT cells were cultured at 1×10⁻⁶ cells / year. 4Cells were seeded at a density of 6 / 100 μL in 96-well plates and DMEM medium was added. Six replicates were set up for each group. The control group was not seeded and the same volume of medium was added. After 24 hours of culture, the model control group and sample groups were incubated using UV microscopy for 70 min. The old medium was then discarded. Fresh medium was added to the control and model wells, while the sample groups (Examples 1-3 and Comparative Examples 1-7) were incubated with medium containing the samples. After another 18 hours of culture, the old medium was discarded, and 100 μL of ATP was added to each well. The plates were lysed at 37°C for 1 h. The supernatant was transferred to a microplate, and the RLU value was measured using a chemiluminescence analyzer. Results were analyzed using SPSS 27.0 statistical software. Data are expressed as mean ± standard deviation. One-way ANOVA was used for comparisons among multiple groups, and t-tests were used for comparisons between two groups. p < 0.05 was considered statistically significant.

[0057] The experimental results are shown in Table 1 below. Compared with the control group, the RLU value of the model group was significantly reduced, indicating successful modeling. Examples 1 to 3 all significantly increased the RLU value of HACAT cells induced by UV damage. Among them, the effect of Example 1 group was the most significant.

[0058] The experimental results from the comparative groups show that the small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu) or Rhodiola rosea exosomes, used alone or in combination with Rhodiola rosea exosomes, significantly increased the RLU value of HACAT cells induced by UV damage, but the effect was not as good as that of groups 1 to 3. However, when the small molecule collagen peptide was used in combination with Rhodiola rosea exosomes, an adverse interaction occurred between the two, and the effect was not as good as that of the small molecule collagen peptide used alone.

[0059] The comparative small molecule collagen peptide HPSAE (His-Pro-Ser-Ala-Glu) alone did not alleviate UV-induced HACAT cell damage, indicating that the activity of small molecule collagen peptides is related to their specific amino acid sequence structure. Furthermore, calendula exosomes alone did not alleviate UV-induced HACAT cell damage, but when used in combination with the small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu) of this invention, the survival rate of HACAT cells reached a level comparable to the control group, and its effect was superior to that produced by combining the small molecule collagen peptide of this invention with Rhodiola rosea exosomes, which inherently improve UV damage. This indicates that the combination of specific small molecule collagen peptides with specific types of plant-derived exosomes produces an unexpectedly significant synergistic effect in alleviating UV-induced HACAT cell damage.

[0060] Table 1: Effect of the composition of the present invention with anti-aging effect on the survival rate of UV-damaged HACAT cells (mean ± standard deviation) Note: Compared with the model group, .

[0061] Example 2: Test on the effect of the composition of the present invention with anti-aging effect on skin hydration. The skin hydration of 55 volunteers (5 in each group) was tested using a Corneometer CM 825 (Courage Khazaka, Germany). A 4 cm × 4 cm test area was marked on the flexor surface of each subject's left and right forearms, and samples (containing, by weight percentage, 2% of Examples 1 to 3 and Comparative Examples 1 to 7, and 98% of conventional cosmetic base ingredients used to prepare the face cream) were applied. The application amount was 3.0 mg / cm². 2 Gently massage until the sample is absorbed. Use a skin moisture meter to measure the skin moisture content before application and 4 hours after application. Calculate the skin moisture increase rate according to the following formula. Take the average skin moisture increase rate of the volunteers who used each sample to obtain the average skin moisture increase rate of that sample.

[0062] Skin moisture content increase rate (%) = (Measured value after application - Baseline value before application) / Baseline value before application x 100% The experimental results are shown in Table 2 below. The data in Table 2 show that using conventional cosmetic base ingredients alone for preparing face creams has poor skin moisturizing effects; after 4 hours, the average increase in skin hydration in the face cream base group volunteers was only 6.25%. The anti-aging composition of this invention, as well as the small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu), both significantly increased the average skin hydration of volunteers when used alone, with the combined form showing even better results. Rhodiola rosea exosomes also have a certain effect on increasing skin hydration in volunteers, but its effect is not as good as the small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu) of this invention, and the effect is actually worse when used in combination with HPTAE (His-Pro-Thr-Ala-Glu).

[0063] In addition, neither the comparative small molecule collagen peptide HPSAE (His-Pro-Ser-Ala-Glu) nor calendula exosomes alone improved skin hydration in volunteers.

[0064] However, the combination of the small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu) of this invention with calendula exosomes can significantly improve the skin hydration of volunteers, and its effect is better than that produced by the combination of the small molecule collagen peptide of this invention with Rhodiola rosea exosomes, which have the effect of improving skin hydration. This indicates that the combination of specific small molecule collagen peptides with specific types of plant-derived exosomes produces an unexpected and significant synergistic effect in improving the skin hydration and enhancing skin moisture in volunteers.

[0065] Table 2: Effects of the composition of the present invention with anti-aging effects on skin hydration Example 3: Anti-wrinkle and firming effect test of the composition of the present invention with anti-aging effect on the skin. Eighty-eight women aged 35-65 were selected as subjects and divided into 11 groups of 8. Before the test, the subjects washed their faces and were placed in a constant temperature and humidity environment (temperature 21°C). 23℃, humidity 55% (65%) Wait 30 minutes, then use VISA CR took facial photographs as initial data. Subjects applied the corresponding face cream (the same cream used in effect example 2) to their faces once a day for 30 consecutive days. On day 0 and day 30, they washed their faces and used a VISA card. CR performed a facial wrinkle test, which was recorded as D0 and D30. The D0 and D30 values ​​for each group are the average values ​​of these values. The instrument software calculated the improvement of facial wrinkles before and after use. The lower the score, the more wrinkles there are.

[0066] The test results are shown in Table 3 below. The data in Table 3 shows that using conventional cosmetic base ingredients for face cream alone has poor anti-wrinkle effects, essentially offering no wrinkle removal. The composition of this invention with skin anti-aging effects, along with the small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu) alone, exhibits significant anti-wrinkle effects, with the combined form showing even better results. Rhodiola rosea exosomes have a slight anti-wrinkle effect, but its effect is less than that of the small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu) of this invention, and the effect worsens when used in combination with HPTAE (His-Pro-Thr-Ala-Glu). Furthermore, the comparative small molecule collagen peptide HPSAE (His-Pro-Ser-Ala-Glu) or calendula exosomes alone have no anti-wrinkle effect. However, the combination of the small molecule collagen peptide HPTAE (His-Pro-Thr-Ala-Glu) of this invention with calendula exosomes can significantly enhance the anti-wrinkle effect of small molecule collagen, and its effect is better than that produced by the combination of the small molecule collagen peptide of this invention with Rhodiola rosea exosomes, which have a slight anti-wrinkle effect. This indicates that the combination of specific small molecule collagen peptides with specific types of plant-derived exosomes produces an unexpected and significant synergistic effect in anti-wrinkle and anti-aging.

[0067] Table 3: Test results of the anti-wrinkle effect of the composition of the present invention with skin anti-aging effect In summary, the experimental results show that by combining small molecule collagen peptides and calendula exosomes in a specific ratio in the composition of the present invention, the composition of the present invention has synergistic effects in moisturizing and repairing the skin barrier, as well as anti-wrinkle and firming. When applied to cosmetics, it can promote the reconstruction of the stratum corneum and barrier function, improve skin hydration and water resistance, reduce wrinkles and make the skin firmer, and has an excellent user experience.

[0068] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A composition having anti-aging effects on the skin, characterized in that: The composition comprises small molecule collagen peptides and calendula exosomes, wherein the weight ratio of the small molecule collagen peptides to calendula exosomes is 1~10:10~1.

2. The composition according to claim 1, characterized in that: The weight ratio of the small molecule collagen peptides to calendula exosomes is 1~8:8~1.

3. The composition according to claim 1, characterized in that: The small molecule collagen peptide was isolated and identified from the enzymatic hydrolysis product of deep-sea cod skin, and the amino acid sequence of the small molecule collagen peptide is HPTAE (His-Pro-Thr-Ala-Glu).

4. The composition according to claim 3, characterized in that: The small molecule collagen peptides were prepared by solid-phase synthesis.

5. The composition according to claim 1, characterized in that: The calendula exosomes have a double-membrane structure under transmission electron microscopy, mostly appearing as concave discs, and the particle size of the calendula exosomes is 50-300 nm.

6. The composition according to claim 5, characterized in that: The method for preparing the calendula exosomes includes the following steps: (1) After washing the fresh calendula flowers with pure water, add an appropriate amount of PBS to soak them. After soaking for a long time, crush them in a blender and filter them through a gauze to obtain filtrate a. (2) Centrifuge filtrate a at 1,000 g for 10-15 min to obtain supernatant a, and centrifuge supernatant a at 3,000 g for 30-40 min to obtain supernatant b; (3) The supernatant b was concentrated by centrifugation at 3,500 g for 30-40 min using a 100 kDa ultrafiltration tube to obtain supernatant concentrate c. Supernatant concentrate c was then centrifuged at 10,000 g for 30-40 min to obtain supernatant concentrate d. (4) The supernatant concentrate d was filtered through a 0.45 μm filter membrane to obtain filtrate b; (5) The filtrate b is centrifuged at 100,000~120,000 g for 60~90 min to obtain a precipitate, which is the calendula exosome.

7. Use of the composition having anti-aging effects according to any one of claims 1 to 6 in the preparation of cosmetics having moisturizing and skin barrier repairing effects as well as anti-wrinkle and firming effects.

8. The use according to claim 7, characterized in that: The cosmetics mentioned are toners, lotions, serums, creams, ointments, masks, or freeze-dried powders.

9. The use according to claim 7, characterized in that: The cosmetic product has a protective effect against UV-induced cell damage, possesses antioxidant activity, repairs damaged cells, protects the skin barrier, and delays skin aging.

10. The use according to claim 7, characterized in that: The composition is present in the cosmetic product at a concentration of 1%-5% by weight.

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