Bicyclic peptide compositions and uses thereof

Abstract

A bicyclic peptide composition and its application are disclosed, relating to the field of peptide technology. The cyclic peptide composition comprises component X and component Y, wherein component X is cyclic tetrapeptide-24-aminocyclohexanecarboxylate, and component Y is selected from one or more of the following cyclic peptides: cyclic hexapeptide-8, cyclic octapeptide-69, Cyclo-(Gly-His-Lys-Lys), Cyclo-(Arg-Arg-Gln-D-Met-Glu-Glu), Cyclo-(Pro-Lys-Glu-Lys), and Cyclo-(Val-Leu-Gln-Trp-Val-Lys). Specifically, the cyclic peptide composition and its use in the preparation of cosmetics are discussed.

Description

Technical Field

[0001] This disclosure relates to the field of peptide technology, and in particular to a bicyclic peptide composition and its applications. Background Technology

[0002] As the body's external barrier tissue, the skin is constantly exposed to multiple exogenous factors such as ultraviolet radiation, environmental pollution, climate change, and daily cleansing, making it prone to abnormal keratin metabolism, impaired barrier function, roughness, dryness, dullness, and decreased elasticity. With age, the skin's renewal capacity declines, and the state of the epidermal barrier and the appearance of the skin change to varying degrees. Therefore, research on compositions for skin care, skin condition improvement, and cosmetic applications has always been a focus of attention.

[0003] In existing technologies, cosmetics and skin care products typically employ a combination of acids, vitamins, plant extracts, peptides, and other functional ingredients to meet application needs such as moisturizing, repairing, smoothing skin, improving skin tone, and mitigating the appearance of aging skin. Among these, peptides have been widely used in product development due to their strong targeting and good biocompatibility. For example, cyclic tetrapeptide-24-aminocyclohexanecarboxylate has been found in cosmetic and skin care technologies and can be effectively used in skin care and beauty, fully utilizing its functions. However, single cyclic peptide ingredients often only target one aspect and cannot simultaneously address multiple skin problems mentioned above.

[0004] Furthermore, although there is some research foundation in peptide formulation, the synergistic effects between different peptide components are highly selective, and not every combination will produce a beneficial effect. The compatibility, stability, mildness, and user experience of different active ingredients in the same formulation system usually require comprehensive balancing. At the same time, existing compositions still require further research in terms of application and efficacy for issues such as rough skin, dullness, changes in skin barrier function, and signs of aging. Therefore, developing novel compositions suitable for skin care and cosmetic applications remains of practical significance. Summary of the Invention

[0005] This disclosure relates to a bicyclic peptide composition and its application, wherein the cyclic peptide composition has effects such as skin care or mucous membrane care.

[0006] On the one hand, the present disclosure provides a cyclic peptide composition, which comprises component X and component Y. Component X is cyclo-tetrapeptide-24-aminocyclohexanecarboxylate, and component Y is selected from one or more of the following cyclic peptides: cyclo-hexapeptide-8, cyclo-octapeptide-69, Cyclo-(Gly-His-Lys-Lys), Cyclo-(Arg-Arg-Gln-D-Met-Glu-Glu), Cyclo-(Pro-Lys-Glu-Lys), Cyclo-(Val-Leu-Gln-Trp-Val-Lys).

[0007] Cyclo-tetrapeptide-24-aminocyclohexanecarboxylate is an active peptide raw material registered and filed by Shenzhen Vicky Technology Co., Ltd. with the National Medical Products Administration. Its filing number is National Cosmetic Original Preparation No. 20240043, and it is denoted as cyclic peptide A in the present disclosure.

[0008] Cyclo-(Gly-His-Lys-Lys) has been publicly recorded in a Chinese patent application (publication number CN118146299A). Its chemical structural formula is shown in formula (I), and it is denoted as cyclic peptide B in the present disclosure. In the present disclosure, cyclic peptide B can be prepared without ambiguity according to the methods in the publicly available literature (such as the Chinese patent application with publication number CN118146299A).

[0009] (I).

[0010] Cyclo-(Arg-Arg-Gln-D-Met-Glu-Glu) has been publicly recorded in a Chinese patent application (publication number CN119978070A). Its chemical structural formula is shown in formula (II), and it is denoted as cyclic peptide C in the present disclosure. In the present disclosure, cyclic peptide C can be prepared without ambiguity according to the methods in the publicly available literature (such as the Chinese patent application with publication number CN119978070A).

[0011] (II).

[0012] Cyclo-hexapeptide-8 is an active peptide raw material registered and filed by Shenzhen Vicky Technology Co., Ltd. with the National Medical Products Administration. Its filing number is National Cosmetic Original Preparation No. 20250151, and it is denoted as cyclic peptide D in the present disclosure.

[0013] Cyclo-octapeptide-69 is an active peptide raw material registered and filed by Shenzhen Vicky Technology Co., Ltd. with the National Medical Products Administration. Its filing number is National Cosmetic Original Preparation No. 20250125, and it is denoted as cyclic peptide E in the present disclosure.

[0014] Cyclo-(Pro-Lys-Glu-Lys) has been disclosed in Chinese invention patent application (publication number CN118240006A), and its chemical structure is shown in formula (III). In this disclosure, it is referred to as cyclic peptide F. In this disclosure, cyclic peptide F can be prepared without question according to the methods described in published literature (e.g., Chinese invention patent application with publication number CN118240006A).

[0015] (III).

[0016] Cyclo-(Val-Leu-Gln-Trp-Val-Lys) has been disclosed in Chinese invention patent (publication number CN120058859A), and its chemical structural formula is shown in formula (IV). In this disclosure, it is referred to as cyclic peptide G. In this disclosure, cyclic peptide G can be prepared without question according to the methods described in published literature (e.g., Chinese invention patent publication number CN120058859A).

[0017] (IV).

[0018] The inventors of this disclosure have surprisingly discovered that when cyclic peptide A is compounded with cyclic peptides B, C, D, E, F, or G, it produces a synergistic effect, significantly removing keratinocytes that fail to shed normally from the skin surface, promoting the expression of multiligand proteoglycan-1, and inhibiting the formation of advanced glycation end products (AGEs). This results in increased skin smoothness, repair of the skin barrier, enhanced epidermal cohesion or dermal-epidermal junction, reduced skin damage from glycation reactions, and improved dullness and / or brightened skin tone. Therefore, compounding cyclic peptide A with cyclic peptides B, C, D, E, F, or G in component Y yields a composition product that simultaneously possesses exfoliating, repairing, and anti-glycation effects.

[0019] In some embodiments, the concentration ratio of component X to component Y is (0.5~2):1.

[0020] In some embodiments, the concentration ratio of component X to component Y is 1:1, 2:1, or 1:2.

[0021] In another aspect of this disclosure, a cosmetic product is provided, comprising an effective amount of the above-described cyclic peptide composition, and at least one additional ingredient.

[0022] "Effective amount" means an amount of the cyclic peptide composition of this disclosure that is non-toxic but sufficient to provide the desired effect. The cyclic peptide composition of this disclosure is used in the cosmetics of this disclosure at an effective concentration to obtain the desired effect. In some embodiments, the concentration of the cyclic peptide composition is between 0.00000001% (by weight) and 20% (by weight) relative to the total weight of the cosmetic; in some embodiments, the concentration of the cyclic peptide composition is between 0.000001% (by weight) and 15% (by weight) relative to the total weight of the cosmetic; in some embodiments, the concentration of the cyclic peptide composition is between 0.0001% (by weight) and 10% (by weight) relative to the total weight of the cosmetic; in some embodiments, the concentration of the cyclic peptide composition is between 0.0001% (by weight) and 5% (by weight) relative to the total weight of the cosmetic.

[0023] In some embodiments, the dosage form of the cosmetic includes ointment, cream, emulsion, aqueous solution, oil, gel, powder, tablet, mud, patch, film, aerosol, spray, lyophilized preparation or nano-preparation.

[0024] Another aspect of this disclosure provides the use of the above-described cyclic peptide composition in the preparation of a composition for caring for skin or mucous membranes.

[0025] Another aspect of this disclosure provides the use of the above-described cyclic peptide composition in the preparation of compositions for exfoliation, repair, or anti-glycation.

[0026] In another aspect of this disclosure, there is provided the use of the above-described cyclic peptide composition in the preparation of a composition for removing keratinocytes that have failed to shed normally from the skin surface; or in the preparation of a composition for increasing skin smoothness; or in the preparation of a composition for repairing the skin barrier; or in the preparation of a composition for promoting the expression of multiligand proteoglycan-1; or in the preparation of a composition for enhancing epidermal cohesion or dermal-epidermal junction; or in the preparation of a composition for inhibiting the formation of advanced glycation end products; or in the preparation of a composition for improving dull skin and / or brightening skin tone.

[0027] Another aspect of this disclosure provides the use of the above-described cyclic peptide composition in the preparation of cosmetics.

[0028] Another aspect of this disclosure provides a cosmetic method for non-therapeutic purposes, comprising using the above-described cyclic peptide composition or the above-described cosmetic on the skin.

[0029] In this disclosure, the term "skin" should be understood as comprising its multiple layers, from the uppermost layer or stratum corneum to the lowermost layer or subcutaneous tissue, including both ends. These layers are composed of different types of cells, such as keratinocytes, fibroblasts, melanocytes, and / or adipocytes.

[0030] The term "skin care" refers to the maintenance and care of the skin to improve its condition, making it delicate, smooth, soft, and healthy.

[0031] The term "keratinocytes that fail to shed normally" refers to terminally differentiated keratinocytes in the stratum corneum of human epidermis that fail to shed in an orderly and physiological manner within the normal epidermal renewal cycle of healthy skin due to abnormal skin metabolism rate, impaired desmosome degradation, damaged barrier function, or the influence of internal and external factors. As a result, these keratinocytes become abnormally retained and accumulated on the skin surface and in the hair follicle infundibulum, leading to objectively verifiable changes in skin condition such as rough skin, blocked hair follicle openings, and thickened stratum corneum.

[0032] This disclosure has the following advantages and effects: 1. The cyclic peptide composition disclosed herein can effectively remove keratinocytes that have not shed normally from the skin surface, thereby achieving the effect of exfoliation and increasing skin smoothness.

[0033] 2. The cyclic peptide composition disclosed herein can effectively promote the expression of multiligand proteoglycan-1, thereby enhancing epidermal cohesion or dermal-epidermal junction, improving skin barrier function, and promoting skin regeneration and repair, thus having a repairing effect.

[0034] 3. The cyclic peptide composition disclosed herein can effectively inhibit the formation of advanced glycation end products, reduce the damage of glycation reaction to the skin, and help improve dull and sallow skin, brighten skin tone, delay skin aging, and has anti-aging and anti-glycation effects. Detailed Implementation

[0035] To make the objects, features, and advantages of this disclosure more apparent and understandable, the disclosure will be further described in detail below with reference to embodiments. Obviously, the described embodiments are only some, not all, of the embodiments of this disclosure. All other embodiments obtained by those skilled in the art based on the embodiments of this disclosure without inventive effort are within the scope of protection of the appended claims.

[0036] Unless otherwise specified, all experimental reagents and materials used in this disclosure are commercially available.

[0037] Cyclic peptide A, cyclic peptide D, and cyclic peptide E were purchased from Shenzhen Vikitech Co., Ltd.

[0038] The structure of Segetalin B is Cyclo-(Gly-Val-Ala-Trp-Ala). In this paper, Segetalin B can be prepared without question according to methods known from the literature. Segetalin B can be prepared according to the literature of Hiroshi Morita et al. (Segetalins B, C and D, Three New Cyclic Peptides From...). Vaccaria segetalis The methods known in [J]. Tetrahedron. 51:21, 6003-6014) are used to obtain information from plants. V. segetalis It is obtained by extraction and separation from the seeds of *Vaccaria segetalis*. Furthermore, those skilled in the art can also obtain *Vaccaria segetalis* cyclic peptide B using conventional solid-phase synthesis methods. *Vaccaria segetalis* cyclic peptide B is referred to as cyclic peptide H in this disclosure.

[0039] The concentrations of each component in the composition of the embodiments of this disclosure are as follows: Composition Cyclic Peptide A Cyclic peptide B Cyclic Peptide C Cyclic Peptide D Cyclic Peptide E Cyclic peptide F Cyclic peptide G Cyclic peptide H Composition 1 1 1 / / / / / / Composition 2 2 1 / / / / / / Composition 3 1 2 / / / / / / Composition 4 1 / 1 / / / / / Composition 5 2 / 1 / / / / / Composition 6 1 / 2 / / / / / Composition 7 1 / / 1 / / / / Composition 8 2 / / 1 / / / / Composition 9 1 / / 2 / / / / Composition 10 1 / / / 1 / / / Composition 11 2 / / / 1 / / / Composition 12 1 / / / 2 / / / Composition 13 1 / / / / 1 / / Composition 14 2 / / / / 1 / / Composition 15 1 / / / / 2 / / Composition 16 1 / / / / / 1 / Composition 17 2 / / / / / 1 / Composition 18 1 / / / / / 2 / Composition 19 1 / / / / / / 1 Composition 20 2 / / / / / / 1 Composition 21 1 / / / / / / 2 Note: " / " indicates that the substance is not present. When the concentration of the composition is 50 ppm, such as cyclic peptide A:cyclic peptide B = 1:1, it means that cyclic peptide A is 25 ppm and cyclic peptide B is 25 ppm.

[0040] Example 1: In vitro exfoliation test

[0041] 1.1 Reagents and Materials Bama fragrant pig skin, PBS, NaOH solution, HCl solution, and BCA protein quantification kit.

[0042] 1.2 Instruments Franz diffusion cell, magnetic stirrer, transdermal moisture loss analyzer, clean bench, and ELISA reader.

[0043] 1.3 Sample to be tested Experimental group: Cyclic peptide A, Cyclic peptide B, Cyclic peptide C, Cyclic peptide D, Cyclic peptide E, Cyclic peptide F, Cyclic peptide G, Cyclic peptide H, Compositions 1-2, Compositions 4-5, Compositions 7-8, Compositions 10-11, Compositions 13-14, Compositions 16-17, Compositions 19-20, dissolved in PBS, with a test concentration of 100 ppm.

[0044] Control group: PBS.

[0045] 1.4 Experimental Methods a) External exfoliation using Bama miniature pig skin, approximately 25mm in diameter and 4.9cm in area. 2 .

[0046] b) Pigskin Integrity Check: Preheat the water bath to 32±1℃ and stabilize the temperature. Install and fix the pigskin between the supply and receiving chambers of the Franz diffusion cell, with the stratum corneum facing the supply chamber and the dermis facing the receiving chamber. Add receiving solution to the receiving chamber through the Franz diffusion cell's replenishment tube, removing air bubbles between the receiving solution and the skin to ensure complete contact between the pigskin and the receiving solution. Turn on the magnetic stirrer at 300 rpm and allow the Franz diffusion cell system to equilibrate under these conditions for 30 minutes. Measure the transepidermal water loss (TEWL) value on the stratum corneum of the pigskin using a transepidermal water loss (TEWL) meter. A TEWL value <15 g / m² is acceptable. 2 Only skin with an integrity level of 6 h can be used for testing. The Bama miniature pig skin used in this test met the integrity requirements.

[0047] c) Fix the Bama fragrant pig skin between the receiving chamber and the supply chamber, take 200 μL of the sample solution to be tested into the supply chamber, use 0.01 M PBS buffer as the receiving chamber system, stir at 300 rpm at 32 °C, and incubate for 24 h.

[0048] d) Place the finger cot over the glass rod, secure it with a rubber band, and place it in the supply chamber to rub the pigskin. After rubbing for 2 minutes, add 300 μL of PBS to wash the supply chamber and collect the keratinocytes shed from the skin surface.

[0049] e) Centrifuge to collect the cell pellet, resuspend the above keratinocytes in 50 μL of deionized water, add 25 μL of 12 mol / L NaOH solution to the liquid, boil in a water bath for 30 min to lyse the keratinocytes, and finally add 25 μL of 12 mol / L HCl solution to neutralize the above lysate.

[0050] f) The total protein concentration of the lysate was determined using the BCA protein quantification kit.

[0051] 1.5 Experimental Results The stratum corneum consists of approximately 10-15 layers of nucleated dead skin cells (keratinocytes), whose main components are proteins and lipids. Under certain environmental conditions, such as dry, cold winters or spring with frequent sandstorms, these keratinocytes can easily accumulate excessively. Timely and moderate exfoliation not only helps improve skin smoothness but also promotes the penetration and absorption of skincare products. In this experiment, test samples were used to treat the skin surface, and the shed keratinocytes were collected for analysis. Since the exfoliation process involves the shedding of keratinocytes, and the number of keratinocytes is positively correlated with the total protein content within them, the number of shed keratinocytes can be indirectly quantitatively characterized by measuring changes in the content of shed protein, thereby analyzing the exfoliating effect of the disclosed cyclic peptide composition. The results of the test samples' influence on the shed keratinocytes on the skin surface are shown in Table 1.

[0052] Table 1. Results of the effect of the test samples on exfoliated keratinocytes on the skin surface (Mean±SD) Test group Relative protein loss (%) control group 100.00±1.04 Cyclic peptide A group 114.29±3.41*** Cyclic peptide B group 111.94±5.20** Cyclic peptide C group 115.84±4.14*** Cyclic peptide D group 104.99±1.97** Cyclic peptide E group 108.66±7.90 Cyclic peptide F group 110.13±3.26** Cyclic peptide G group 108.66±2.39** Cyclic peptide H group 103.19±3.04 Group 1 of the composition 133.55±6.43*** Composition 2 groups 129.29±6.87*** Composition 4 groups 134.08±7.96*** 5 groups of compositions 137.59±7.11*** 7 groups of compositions 140.68±5.75*** 8 groups of compositions 137.49±5.62*** 10 groups of compositions 138.76±6.93*** 11 groups of compositions 138.98±7.57*** 13 groups of compositions 135.25±1.77*** 14 groups of compositions 137.70±5.73*** 16 groups of compositions 135.57±2.05*** 17 groups of compositions 125.35±3.92*** 19 compositions 109.14±4.87* 20 groups of compositions 104.25±2.21* Note: Compared with the control group, * P <0.05,** P <0.01, *** P <0.001.

[0053] The experimental results showed that, compared with the control group, the relative protein shedding in the experimental groups, except for the cyclic peptide E group and the cyclic peptide H group, was significantly increased. This demonstrates that the samples in the other experimental groups, except for cyclic peptide E and cyclic peptide H, can effectively remove keratinocytes that have not shed normally from the skin surface, thus exhibiting exfoliation effects. Specifically, the relative protein shedding in composition groups 1-2 was significantly better than that in cyclic peptide A and cyclic peptide B groups; the relative protein shedding in composition groups 4-5 was significantly better than that in cyclic peptide A and cyclic peptide C groups; the relative protein shedding in composition groups 7-8 was significantly better than that in cyclic peptide A and cyclic peptide D groups; the relative protein shedding in composition groups 10-11 was significantly better than that in cyclic peptide A and cyclic peptide E groups; the relative protein shedding in composition groups 13-14 was significantly better than that in cyclic peptide A and cyclic peptide F groups; and the relative protein shedding in composition groups 16-17 was significantly better than that in other groups. The results were significantly better than those of the cyclic peptide A and cyclic peptide G groups, indicating that compared to cyclic peptides A, B, C, D, E, F, and G, cyclic peptide A, when combined with specific cyclic peptides B, C, D, E, F, and G, exhibited superior exfoliation effects. This demonstrates that cyclic peptide A, when combined with cyclic peptides B, C, D, E, F, and G, produces a synergistic effect, more significantly removing keratinocytes that have not shed normally from the skin surface, thereby further enhancing its exfoliation efficacy. In contrast, the relative protein shedding rates of compositions 19-20 were 109.14% and 104.25%, respectively, which, although significantly different from the control group, were significantly lower than those of the cyclic peptide A group. Compared to the cyclic peptide H group, no statistically significant difference was observed, and the synergistic effect was not met, indicating that the combination of cyclic peptide A and cyclic peptide H did not produce a synergistic effect.

[0054] Further comparison of the effects of different concentration ratios on the relative amount of protein detachment revealed that, in the combination of cyclic peptide A and cyclic peptide G, the relative amount of protein detachment in composition group 16 with a concentration ratio of 1:1 was 135.57%, significantly higher than that in composition group 17 with a concentration ratio of 2:1. This indicates that under the experimental conditions and within the range of concentration ratios examined, a concentration ratio of 1:1 for cyclic peptide A and cyclic peptide G showed relatively better performance in the exfoliation efficacy evaluation index and had the best synergistic effect.

[0055] In summary, the cyclic peptide composition disclosed herein can be used to remove keratinocytes that have not shed properly from the skin surface, thereby achieving the effect of exfoliation and increasing skin smoothness.

[0056] Example 2: Multiligand proteoglycan-1 content test

[0057] 2.1 Reagents and Materials Syndecan-1 ELISA kit, RIPA extract, BCA protein quantification kit, trypsin, fetal bovine serum, DMEM medium, phosphate buffer.

[0058] 2.2 Instruments CO2 incubator, ELISA reader, and clean bench.

[0059] 2.3 Cell lines Human keratinocytes (HaCaT).

[0060] 2.4 Sample to be tested Experimental group: Cyclic peptide A, cyclic peptide B, cyclic peptide C, cyclic peptide D, cyclic peptide E, cyclic peptide F, cyclic peptide G, composition 3, composition 6, composition 9, composition 12, composition 15, and composition 18 were dissolved in PBS and tested at concentrations of 10 ppm and 50 ppm. Cyclic peptide H and composition 21 were dissolved in PBS and tested at concentrations of 10 ppm and 50 ppm.

[0061] Blank control group: PBS.

[0062] UV group: PBS, plus UV radiation.

[0063] 2.5 Experimental Methods Take one flask of cells in the exponential growth phase, add 0.25% trypsin digestion solution, digest to detach the adherent cells, and count (1~4) × 10⁻⁶ cells. 5 Cells were prepared at a density of 10 cells / mL to form a cell suspension. The cells were then seeded into 12-well plates and incubated in a CO2 incubator for 24 hours.

[0064] Complete culture medium was aspirated from the wells. For the blank control group, 200 μL of PBS was added, and the culture medium was replenished to 800 μL without UVB irradiation. For the UV group and experimental group, 200 μL of PBS was added, and the mixture was irradiated under a UVB lamp for 15 min (irradiation intensity: 80-90 μW / cm²). 2 Irradiation dose: 80 mJ / cm 2 After irradiation for 15 min, the PBS solution was aspirated. 200 μL of PBS was added to the UV group, and the culture medium was replenished to 800 μL. 200 μL of samples of different concentrations were added to the experimental groups, and the culture medium was replenished to 800 μL. The blank control group, UV group, and experimental groups were incubated at 37℃ in a 5% CO2 incubator for 48 h.

[0065] After culture, cells were collected, centrifuged to obtain the cell pellet, and RIPA lysis buffer was added. The pellet was vortexed three times (30 s / time, 3 min interval) and centrifuged at 12000 rpm for 10 min. The supernatant was collected and analyzed according to the multiligand proteoglycan-1 ELISA instruction manual.

[0066] 2.6 Experimental Results In keratinocytes, multiligand proteoglycan-1 (SDC-1), a major transmembrane heparan sulfate proteoglycan, plays a crucial role in maintaining the structural and functional integrity of the epidermis. SDC-1 not only participates in cell-extracellular matrix adhesion but also regulates keratinocyte proliferation, differentiation, and migration, and is involved in the interaction between cells and the microenvironment, as well as the regulation of physiological processes related to skin repair and regeneration. In this experiment, keratinocytes treated with test samples after UV irradiation were examined to determine the SDC-1 content in the keratinocytes, aiming to ascertain whether the cyclic peptide composition disclosed herein could promote SDC-1 expression in keratinocytes. The effects of the test samples on SDC-1 expression in HaCaT cells are shown in Table 2.

[0067] Table 2. Effects of the test samples on the relative expression level of SDC-1 in HaCaT cells (Mean±SD) Note: Compared with the blank control group ## P <0.01; compared with the UV group, * P <0.05,** P <0.01, *** P <0.001.

[0068] The results showed that, compared with the blank control group, the relative expression level of SDC-1 in the UV group was significantly reduced, indicating successful modeling. Compared with the UV group, except for the cyclic peptide B group, the relative expression level of SDC-1 in the experimental groups was significantly increased, proving that the other experimental group samples, except for cyclic peptide B, could effectively promote the expression of SDC-1 in HaCaT cells. Moreover, the relative expression level of SDC-1 increased with increasing concentration, indicating that the experimental group samples were the direct factor affecting SDC-1 expression, and there was a concentration-dependent relationship between SDC-1 expression and the experimental group samples.

[0069] Further analysis revealed that the relative expression levels of SDC-1 in HaCaT cells of the 10 ppm and 50 ppm composition groups (Group 3) were 171.51% and 200.06%, respectively, both significantly higher than those of the cyclic peptide A and cyclic peptide B groups. This indicates that the combination of cyclic peptide A and cyclic peptide B produced a synergistic effect, significantly promoting SDC-1 expression. Similarly, the relative expression levels of SDC-1 in HaCaT cells of the 10 ppm and 50 ppm composition groups (Group 6) were 184.15% and 224.80%, respectively, both significantly higher than those of the cyclic peptide A and cyclic peptide C groups. This indicates that the combination of cyclic peptide A and cyclic peptide C produced a synergistic effect, significantly promoting SDC-1 expression. Finally, the relative expression levels of SDC-1 in HaCaT cells of the 10 ppm and 50 ppm composition groups (Group 9) were 168.71% and 169.77%, respectively, both significantly higher than those of the cyclic peptide A and cyclic peptide D groups. This indicates that the combination of cyclic peptide A and cyclic peptide D produced a synergistic effect, significantly promoting SDC-1 expression. In HaCaT cells of groups 12 (containing 10 ppm and 50 ppm compositions), the relative expression levels of SDC-1 were 160.74% and 179.95%, respectively, both significantly higher than those of groups cyclic peptide A and cyclic peptide E, indicating a synergistic effect between cyclic peptide A and cyclic peptide E, which significantly promoted SDC-1 expression. Similarly, in groups 15 (containing 10 ppm and 50 ppm compositions), the relative expression levels of SDC-1 were 157.85% and 194.84%, respectively, both significantly higher than those of groups cyclic peptide A and cyclic peptide F, indicating a synergistic effect between cyclic peptide A and cyclic peptide F, which significantly promoted SDC-1 expression. Finally, in groups 18 (containing 10 ppm and 50 ppm compositions), the relative expression levels of SDC-1 were 228.67% and 234.62%, respectively, both significantly higher than those of groups cyclic peptide A and cyclic peptide G, indicating a synergistic effect between cyclic peptide A and cyclic peptide G, which significantly promoted SDC-1 expression. In contrast, although the relative expression level of SDC-1 in composition 21 was higher than that in the UV group, it was significantly lower than that in the cyclic peptide A group and the cyclic peptide H group at the same concentration, and it did not meet the standard for synergistic effect. This suggests that the combination of cyclic peptide A and cyclic peptide H failed to produce a synergistic effect, which also proves that the synergistic effect of the cyclic peptide composition disclosed in this paper has component selectivity.

[0070] In summary, the cyclic peptide composition disclosed herein can promote the expression of multiligand proteoglycan-1, thereby enhancing epidermal cohesion or dermal-epidermal junction, improving skin barrier function, and promoting skin regeneration and repair, thus having a repairing effect.

[0071] Example 3 Anti-glycation test

[0072] 3.1 Reagents and Materials Phosphate-buffered saline (PBS), pyrrolizaldehyde (MGO), and bovine serum albumin (BSA).

[0073] 3.2 Instruments Clean bench, enzyme-linked immunosorbent assay (ELISA) reader.

[0074] 3.3 Sample to be tested Experimental group: Cyclic peptide A, Cyclic peptide B, Cyclic peptide C, Cyclic peptide D, Cyclic peptide E, Cyclic peptide F, Cyclic peptide G, Cyclic peptide H, Composition 3, Composition 6, Composition 9, Composition 12, Composition 15, Composition 18, Composition 21, dissolved in PBS, with a test concentration of 100 ppm.

[0075] Control group: PBS.

[0076] 3.4 Experimental Methods Weigh 100 mg of bovine serum albumin and add it to 10 mL of PBS solution. Dissolve completely to prepare a 10 mg / mL BSA solution. Weigh 14.41 mg of acetone and add it to 40 mL of PBS solution. Dissolve completely to prepare a 5 mM MGO solution. Mix the 10 mg / mL BSA solution and the 5 mM MGO solution in 0.2 M phosphate buffer (pH 7.4) to prepare AGEs.

[0077] The experimental group samples were added to the BSA-MGO model at a volume ratio of 1:1 and shaken thoroughly. All reaction mixtures were incubated at 37°C under sterile conditions for 21 days. The control group was prepared by replacing the test samples with the same volume of PBS, with all other procedures performed identically. Before determining the anti-glycation effect, each reaction mixture was diluted 5-fold with 160 μL of PBS buffer to pH 7.4, then transferred to a 96-well plate, and the fluorescence intensity was measured at an excitation wavelength of 340 nm and an emission wavelength of 435 nm. The AGEs inhibition rate of the test samples was calculated using the following formula: AGEs inhibition rate % = (1 - F1 / F0) × 100% Wherein, F1 is the fluorescence value of the sample to be tested, and F0 is the fluorescence value of the control group.

[0078] 3.5 Experimental Results Advanced glycation end products (AGEs) are stable end products formed by the non-enzymatic glycation reaction of proteins with reducing sugars. The amount of AGEs generated, exhibiting fluorescence properties, is positively correlated with their fluorescence intensity. In this experiment, an in vitro AGEs generation model was constructed using BSA and MGO. By detecting changes in fluorescence intensity of different samples, the anti-glycation activity of the disclosed cyclic peptide composition was determined. The results of the influence of the test samples on AGEs generation are shown in Table 3.

[0079] Table 3. Results of the effect of test samples on AGEs formation (Mean ± SD) Test group AGEs inhibition rate (%) control group 0.00±1.19 Cyclic peptide A group 10.06±1.27** Cyclic peptide B group 1.61±1.53 Cyclic peptide C group 13.16±0.86*** Cyclic peptide D group 9.54±1.36** Cyclic peptide E group 15.63±0.76*** Cyclic peptide F group 2.88±2.04 Cyclic peptide G group 13.20±0.51*** Cyclic peptide H group 0.72±1.00 Composition 3 groups 31.07±1.28*** Composition 6 groups 39.65±0.75*** 9 groups of compositions 39.79±0.86*** 12 groups of compositions 36.48±1.17*** 15 groups of compositions 34.54±0.29*** 18 groups of compositions 37.54±0.59*** 21 compositions 3.47±0.84* Note: Compared with the control group, * P<0.05,** P <0.01, *** P <0.001.

[0080] The results showed that, compared with the control group, cyclic peptide A, cyclic peptide C, cyclic peptide D, cyclic peptide E, cyclic peptide G, combination 3, combination 6, combination 9, combination 12, combination 15, combination 18, and combination 21 all significantly inhibited AGEs formation. Cyclic peptide B, cyclic peptide F, and cyclic peptide H did not show significant inhibitory effects on AGEs formation. Among them, the AGEs inhibition rate of combination 3 was 31.07%, significantly better than that of cyclic peptide A and cyclic peptide B, indicating that the combination of cyclic peptide A and cyclic peptide B produced a synergistic effect, which more significantly inhibited AGEs formation. The AGEs inhibition rate of combination 6 was 39.65%, significantly better than that of cyclic peptide A and cyclic peptide C, indicating that the combination of cyclic peptide A and cyclic peptide C produced a synergistic effect, which more significantly inhibited AGEs formation. The AGEs inhibition rate of combination 9 was 39.79%, significantly better than that of cyclic peptide A and cyclic peptide D, indicating that the combination of cyclic peptide A and cyclic peptide D produced a synergistic effect, which more significantly inhibited AGEs formation. The AGEs inhibition rate of composition 12 was 36.48%, significantly better than that of cyclic peptide A and cyclic peptide E, indicating that the combination of cyclic peptide A and cyclic peptide E produced a synergistic effect, which could more significantly inhibit AGEs formation. The AGEs inhibition rate of composition 15 was 34.54%, significantly better than that of cyclic peptide A and cyclic peptide F, indicating that the combination of cyclic peptide A and cyclic peptide F produced a synergistic effect, which could more significantly inhibit AGEs formation. The AGEs inhibition rate of composition 18 was 37.54%, significantly better than that of cyclic peptide A and cyclic peptide G, indicating that the combination of cyclic peptide A and cyclic peptide G produced a synergistic effect, which could more significantly inhibit AGEs formation. In contrast, the inhibition rate of composition 21 was only 3.47%, which was not only far lower than the aforementioned specific combination compositions, but also significantly lower than the level of cyclic peptide A monomer (…). P The cyclic peptide A and cyclic peptide H were found to have a synergistic effect (<0.01), and the synergistic effect was not achieved. Therefore, it was concluded that the combination of cyclic peptide A and cyclic peptide H did not produce a synergistic effect, and may even have produced an antagonistic effect.

[0081] In summary, the cyclic peptide composition disclosed herein can effectively inhibit the formation of advanced glycation end products, thereby reducing the damage of glycation reactions to the skin, improving dull and sallow skin, delaying skin aging, brightening skin tone, and having anti-aging and anti-glycation effects.

[0082] Example 4

[0083] An essence is prepared through the following steps, and the specific formula is shown in Table 4 below: Table 4 According to the formula dosage, add all materials of phase A to the mixing pot, stir and heat to 80-85℃, keep warm for 30-35 minutes, start stirring and cooling down to 60-65℃, add all materials of phase B, continue stirring and cooling down to 35-40℃, add materials of phases C and D, stir for 10-15 minutes, and the product is ready.

[0084] In this disclosure, relational terms such as "first" and "second" are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or terminal device that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or terminal device. Without further limitation, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or terminal device that includes said element.

[0085] While specific embodiments of this disclosure have been described for illustrative purposes, various modifications or alterations can be made by those skilled in the art without departing from the spirit and scope of this disclosure. All such modifications or alterations should fall within the scope of the appended claims.

Claims

1. A cyclic peptide composition, characterized in that, The cyclic peptide composition comprises component X and component Y, wherein component X is cyclic tetrapeptide-24-aminocyclohexanecarboxylate, and component Y is selected from one or more of the following cyclic peptides: cyclic hexapeptide-8, cyclic octapeptide-69, Cyclo-(Gly-His-Lys-Lys), Cyclo-(Arg-Arg-Gln-D-Met-Glu-Glu), Cyclo-(Pro-Lys-Glu-Lys), and Cyclo-(Val-Leu-Gln-Trp-Val-Lys).

2. The cyclic peptide composition according to claim 1, characterized in that, The concentration ratio of component X to component Y is (0.5~2):

1.

3. The cyclic peptide composition according to claim 1, characterized in that, The concentration ratio of component X to component Y is 1:1, 2:1, or 1:

2.

4. A cosmetic product, characterized in that, The composition comprises an effective amount of the cyclic peptide composition according to any one of claims 1-3, and at least one additional ingredient.

5. The cosmetic product according to claim 4, characterized in that, The dosage forms of the cosmetics include ointments, creams, emulsions, liquids, oils, gels, powders, tablets, muds, patches, films, aerosols, sprays, freeze-dried preparations, or nano-preparations.

6. Use of the cyclic peptide composition according to any one of claims 1-3 in the preparation of compositions for the care of skin or mucous membranes.

7. Use of the cyclic peptide composition according to any one of claims 1-3 in the preparation of compositions for exfoliation, repair or anti-glycation.

8. Use of the cyclic peptide composition according to any one of claims 1-3 in the preparation of a composition for removing keratinocytes that fail to shed normally from the skin surface; or in the preparation of a composition for increasing skin smoothness; or in the preparation of a composition for repairing the skin barrier; or in the preparation of a composition for promoting the expression of multiligand proteoglycan-1; or in the preparation of a composition for enhancing epidermal cohesion or dermal-epidermal junction; or in the preparation of a composition for inhibiting the formation of advanced glycation end products; or in the preparation of a composition for improving dull skin and / or brightening skin tone.

9. Use of the cyclic peptide composition according to any one of claims 1-3 in the preparation of cosmetics.

10. A cosmetic method for non-therapeutic purposes, characterized in that, The method includes using the cyclic peptide composition of any one of claims 1-3, or the cosmetic of claim 4 or 5, on the skin.

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