Polypeptide composition containing moringa seed extract and application of polypeptide composition in skin repair

Abstract

The invention discloses a polypeptide composition containing a moringa seed extract and application of the polypeptide composition in skin repair, and belongs to the technical field of biology, and the polypeptide composition comprises the moringa seed extract, polypeptide SEQ ID NO.1 derived from brasenia schreberi and polypeptide SEQ ID NO.2 derived from mulberries. The composition is clear and definite in component, all the components take effect synergistically through different action mechanisms, and the composition is free of side effects and free of toxic and side effects. And the problem of unpredictability of a complex formula is solved. The moringa seed extract provides a broad-spectrum anti-oxidation and anti-inflammatory basis, and the brasenia schreberi source polypeptide as a membrane protein related peptide interacts with a skin cell membrane, so that the skin permeation effect of effective components such as the moringa seed extract is promoted, and skin cell absorption and repair are accelerated; mulberry source polypeptide has the potential of inhibiting tyrosinase activity and reducing melanogenesis, and pigmentation is inhibited in a targeted mode. Through scientific proportioning and an advanced extraction process, ideal effects are achieved in the aspects of skin repairing, moisturizing, spot fading and the like.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biotechnology, in particular to a polypeptide composition containing moringa seed extract and its application in skin repair. BACKGROUND

[0002] Skin is the outermost organ of the human body composed of many tissues, which directly contacts with the outside world and is easily affected by external factors such as ultraviolet rays, air pollution, chemical substances and physical damage, resulting in a series of problems such as damaged skin barrier, water loss, inflammatory response and pigmentation. Therefore, people's demand for skin repair, moisturizing, anti-aging and the like is also increasing.

[0003] In the prior art, moringa seeds contain rich proteins, vitamins, minerals and polyphenols, and the extract thereof has the effects of antioxidant, anti-inflammatory and moisturizing. There are patents reported, such as CN110420150A, which supplements skin moisture and nutrients by cooperating with other ingredients mother chrysanthemum extract, centella asiatica extract, licorice root extract, tea extract, giant knotweed root extract, rosemary leaf extract and radix scutellariae extract, to moisturize the skin and achieve anti-aging effect. CN113081910A, which combines moringa seeds with numerous ingredients rhizobium gum, trehalose, mucuna seed polysaccharide, white pool flower seed oil, tocopheryl acetate, betaine, gardenia fruit extract, mannose phosphate sodium, mannose, nicotinamide and bisabolol, has multiple effects such as moisturizing, whitening, anti-wrinkle, anti-aging, anti-sensitivity and soothing. As described above, the ingredients in cosmetics are complex and diverse, and the properties of each ingredient are uncertain. In particular, it is unknown whether the combination will cause potential damage to the skin. Moreover, although plant extracts have high safety, they often have problems such as unstable content of active ingredients, large molecular weight and difficulty in absorption, single effect and insufficient effect.

[0004] Therefore, how to provide a new active ingredient composition with clear ingredients, through scientific proportioning and advanced extraction process, to achieve a relatively ideal effect in skin repair, moisturizing, spot lightening and the like, is a technical problem to be solved by those skilled in the art. SUMMARY

[0005] To this end, the present application provides a polypeptide composition containing moringa seed extract and its application in skin repair, to solve the related technical problems in the prior art.

[0006] In order to achieve the above-mentioned purpose, the present application provides the following technical solutions: According to a first aspect of the present application, a polypeptide composition containing moringa seed extract is provided, which comprises the following ingredients by weight: Moringa seed extract 3-12 parts Polypeptide derived from cabomba 0.5-3.5 parts Polypeptide derived from mulberry 1-5 parts The polypeptide derived from cabomba is as shown in SEQ ID NO. 1, and the polypeptide derived from mulberry is as shown in SEQ ID NO. 2.

[0007] Further, the following ingredients are included by weight parts: Moringa seed extract 8 parts Polypeptide derived from cabomba 1.5 parts Polypeptide derived from mulberry 3 parts.

[0008] Further, the extraction method of the polypeptide SEQ ID NO. 1 is as follows: (1) Raw material pretreatment: wash the cabomba sample with pre-cooled distilled water, remove impurities and absorb water with filter paper; (2) Grinding: quickly freeze the cabomba sample in liquid nitrogen, transfer it to a pre-cooled mortar, and grind the frozen cabomba into extremely fine powder; (3) Crude extraction: transfer the ground powder to a beaker containing pre-cooled extraction Tris-HCl buffer, continuously stir on ice for 30-60 min, centrifuge at 3000 rpm at 4°C for 10-15 min, collect the supernatant; centrifuge at 20000 rpm at 4°C for 30-45 min, collect the supernatant; ultracentrifuge at 100000 rpm at 4°C for 30-45 min, collect the precipitate; (4) Fine purification: redissolve the above precipitate in Tris-HCl buffer containing 1% DDM, slowly shake at 4°C, incubate for 8-12 h, ultracentrifuge the incubated mixture at 100000 rpm at 4°C for 60-90 min to obtain the supernatant; (5) Purification: sequentially pass through an affinity chromatography column and a cation exchange chromatography column for chromatography, collect multiple protein peaks and perform dialysis purification, add the polypeptides corresponding to the multiple protein peaks to the company's cosmetics for testing, screen 3 polypeptides with moisturizing and spot-fading repair effects, and finally determine one polypeptide with the best effect for subsequent experiments, which is as shown in SEQ ID NO. 1.

[0009] Further, the extraction method of the polypeptide SEQ ID NO. 2 is as follows: (1) Raw material pretreatment: wash the mulberry sample with pre-cooled distilled water, remove impurities and absorb water with filter paper; (2) Grinding: quickly freeze the mulberry sample in liquid nitrogen, transfer it to a pre-cooled mortar, and grind the frozen mulberry into extremely fine powder; (3) Extraction: transferring the ground powder into a beaker containing pre-cooled extraction Tris-HCl buffer, continuously stirring on ice for 30-60 min, centrifuging at 8000 rpm for 30-45 min at 4℃, collecting the supernatant to obtain mulberry extract; (4) Purification: sequentially passing through an affinity chromatography column and a cation exchange chromatography column for chromatography, collecting multiple protein peaks and performing dialysis purification, adding the multiple polypeptides corresponding to the multiple protein peaks into the cosmetics of the company for testing, screening 5 polypeptides having the effects of moisturizing and spot lightening repair, and finally determining one polypeptide having the best effect for subsequent experiments, and the polypeptide is shown as SEQ ID NO. 2.

[0010] According to the second aspect of the present application, the use of the polypeptide composition containing the extract of moringa oleifera seeds as described above in the preparation of a skin repair medicine or cosmetic is provided.

[0011] Further, the skin repair cosmetic formulation comprises the following components by weight: Polypeptide composition 10-25 parts Humectant 5-10 parts Emollient 7-15 parts Emulsifier 2-5 parts Antioxidant 0.5-1 part Preservative 0.5-1.5 parts Diluted with solvent water.

[0012] Further, the emollient is one or both of cyclopentasiloxane and squalane; and the emulsifier is PEG-10 dimethicone.

[0013] Further, the humectant is one or both of 1,3-propanediol and sodium hyaluronate.

[0014] Further, the antioxidant is one or more of tocopherol acetate, vitamin C and vitamin E.

[0015] Further, the preservative is one or more of hexylene glycol, phenoxyethanol and chlorphenesin.

[0016] The present application has the following advantages: The present application creatively applies three ingredients of Moringa oleifera seed extract, Potamogeton crispus polypeptide and mulberry polypeptide, has clear ingredients and synergistic effects, and solves the unpredictability problem of complex formula. The composition has clear ingredients, and the components have synergistic effects through different mechanisms. The Moringa oleifera seed extract provides a broad-spectrum antioxidant and anti-inflammatory basis, the Potamogeton crispus polypeptide SEQ ID NO. 1 as a membrane protein related peptide interacts with the skin cell membrane, promotes the skin penetration effect of the effective components such as the Moringa oleifera seed extract, and accelerates the absorption and repair of skin cells; the mulberry polypeptide SEQ ID NO. 2 has the potential to inhibit the activity of tyrosinase and reduce melanin production, specifically inhibits pigmentation, and improves the repair effect. The three ingredients complement each other and jointly construct a skin repair system of “anti-oxidation and anti-inflammation-repairing barrier-fading color spots”, and experiments prove that the comprehensive repair effect is ideal. DETAILED DESCRIPTION

[0017] The embodiments of the present application are described below by specific examples, and those skilled in the art can easily understand other advantages and effects of the present application from the disclosure. Obviously, the described examples are part of the examples of the present application, not all. Based on the examples in the present application, all other examples obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0018] In order to better solve the related technical problems existing in the prior art, the present application provides the following technical solutions, aiming to solve the unpredictability problem of complex formula and the poor use effect problem by using ingredients with clear ingredients and synergistic effects.

[0019] According to a first aspect of the present application, a polypeptide composition containing Moringa oleifera seed extract is provided, including the following ingredients by weight: Moringa oleifera seed extract 3-12 parts Polypeptide derived from Potamogeton crispus 0.5-3.5 parts Polypeptide derived from mulberry 1-5 parts The polypeptide derived from Potamogeton crispus is shown as SEQ ID NO. 1, and the polypeptide derived from mulberry is shown as SEQ ID NO. 2.

[0020] Further, the extraction method of the polypeptide SEQ ID NO. 1 is as follows: (1) Raw material pretreatment: wash the Potamogeton crispus sample with pre-cooled distilled water, remove impurities and dry the water with filter paper; (2) Grinding: quickly freeze the Potamogeton crispus sample in liquid nitrogen, transfer it to a pre-cooled mortar, and grind the frozen Potamogeton crispus into extremely fine powder; (3) crude extraction: the ground powder is transferred to a beaker containing pre-cooled extraction Tris-HCl buffer, continuously stirred on ice for 30-60 min, centrifuged at 3000 rpm for 10-15 min at 4℃, and the supernatant is collected; high-speed centrifugation at 20000 rpm for 30-45 min at 4℃, and the supernatant is collected; ultracentrifugation at 100000 rpm for 30-45 min at 4℃, and the precipitate is collected; (4) fine purification: the above precipitate is redissolved in Tris-HCl buffer containing 1% DDM, slowly shaken at 4℃, incubated for 8-12 h, and the incubated mixture is ultracentrifuged at 100000 rpm for 60-90 min at 4℃ to obtain the supernatant; (5) purification: chromatography is sequentially performed through an affinity chromatography column and a cation exchange chromatography column, multiple protein peaks are collected and dialysis purification is performed, multiple polypeptides corresponding to the multiple protein peaks are respectively added to the cosmetics of the company for testing, 3 polypeptides with the effects of moisturizing and freckle repair are screened, and finally one polypeptide with the best effect is determined for subsequent experiments, and the polypeptide is shown as SEQ ID NO. 1.

[0021] Further, the extraction method of the polypeptide SEQ ID NO. 2 is as follows: (1) raw material pretreatment: the mulberry sample is washed with pre-cooled distilled water, impurities are removed, and the water is absorbed with filter paper; (2) grinding: the mulberry sample is quickly frozen in liquid nitrogen and transferred to a pre-cooled mortar, and the frozen mulberry is ground into extremely fine powder; (3) extraction: the ground powder is transferred to a beaker containing pre-cooled extraction Tris-HCl buffer, continuously stirred on ice for 30-60 min, centrifuged at 8000 rpm for 30-45 min at 4℃, and the supernatant is collected to obtain mulberry extract; (4) purification: chromatography is sequentially performed through an affinity chromatography column and a cation exchange chromatography column, multiple protein peaks are collected and dialysis purification is performed, multiple polypeptides corresponding to the multiple protein peaks are respectively added to the cosmetics of the company for testing, 3 polypeptides with the effects of moisturizing and freckle repair are screened, and finally one polypeptide with the best effect is determined for subsequent experiments, and the polypeptide is shown as SEQ ID NO. 2.

[0022] According to the second aspect of the present application, the polypeptide composition containing moringa seed extract as above is used for preparing skin repair drugs or cosmetics.

[0023] Further, the skin repair cosmetic formula includes the following components by weight: Polypeptide composition 10-25 parts Humectant 5-10 parts Emollient 7-15 parts Emulsifier 2-5 parts Antioxidant 0.5-1 part Preservative 0.5-1.5 parts Dilute with solvent water to volume.

[0024] Further, the emollient is one or both of cyclomethicone and squalane; the emulsifier is PEG-10 dimethicone.

[0025] Further, the humectant is one or both of 1,3-propanediol and sodium hyaluronate.

[0026] Further, the antioxidant is one or more of tocopheryl acetate, vitamin C, and vitamin E.

[0027] Further, the preservative is one or more of hexylene glycol, phenoxyethanol, and chlorphenesin.

[0028] In order to better illustrate the technical effects of the present application, the following examples and test examples are provided.

[0029] Preparation Example 1

[0030] Extraction of the polypeptide SEQ ID NO. 1 derived from Potamogeton crispus, as follows: (1) Raw material pretreatment: Prepare fresh Potamogeton crispus by removing stems and retaining leaves, wash the Potamogeton crispus sample with pre-cooled distilled water, remove impurities, and dry the water with filter paper, and reserve for use; (2) Grinding: Place the Potamogeton crispus sample into liquid nitrogen for quick freezing, after 30 s of freezing time, quickly transfer to a pre-cooled mortar, and grind the frozen Potamogeton crispus into extremely fine powder; (3) Crude extraction: Transfer the ground powder into a beaker containing pre-cooled Tris-HCl buffer solution, continuously stir on ice for 50 min, centrifuge at 3000 rpm for 15 min at 4°C, collect the supernatant, and the precipitate is the unground fragments and other impurities; centrifuge the above supernatant at 20000 rpm for 40 min at 4°C, collect the supernatant, and the precipitate is chloroplast and other impurities; ultracentrifuge the above supernatant at 100000 rpm for 40 min at 4°C, collect the precipitate to obtain the crude extract; (4) Fine purification: Redissolve the above precipitate in Tris-HCl buffer solution containing 1% DDM, slowly shake and incubate at 4°C for 12 h, and ultracentrifuge the incubated mixture at 100000 rpm for 90 min at 4°C to obtain the supernatant; (5) Purification: The supernatant was sequentially subjected to affinity chromatography column and cation exchange chromatography column, and multiple protein peaks were collected and dialyzed for purification. The above protein purification process applies AKTA protein purification system, which is a routine technique in the art, and the purification related parameters are not described here. The polypeptides corresponding to the multiple protein peaks were added to the company's cosmetics (brand name: Lv Xun, product name: Su Xin vitality cream) for testing, and 3 polypeptides with moisturizing and spot lightening repair effects were screened, and finally one polypeptide with the best effect was determined for subsequent experiments. The polypeptide was obtained by sequencing, as shown in SEQ ID NO. 1.

[0031] In the above extraction process, it was found that the content of polypeptide SEQ ID NO. 1 in Potamogeton crispus was low, which may be derived from cell membrane related transmembrane protein. In order to obtain sufficient amount of polypeptide for subsequent experiments, Hubei Qiangyao Biotechnology Co., Ltd. was entrusted to biosynthesize, and the purity was not less than 90%.

[0032] Preparation Example 2

[0033] Extraction of polypeptide SEQ ID NO. 2 derived from mulberry, as follows: (1) Raw material pretreatment: fresh mulberries were prepared, and the mulberry samples were washed with pre-cooled distilled water to remove impurities and dried with filter paper for standby; (2) Grinding: the mulberry samples were quickly frozen in liquid nitrogen, and after 1 min of freezing time, they were quickly transferred to a pre-cooled mortar and ground into extremely fine powder; (3) Extraction: the ground powder was transferred to a beaker containing pre-cooled Tris-HCl buffer solution, and continuously stirred on ice for 50 min. After centrifugation at 8000 rpm for 40 min at 4°C, the supernatant was collected to obtain mulberry extract; (4) Purification: sequentially subjected to affinity chromatography column and cation exchange chromatography column, and multiple protein peaks were collected and dialyzed for purification. The above protein purification process applies AKTA protein purification system, which is a routine technique in the art, and the purification related parameters are not described here. The polypeptides corresponding to the multiple protein peaks were added to the company's cosmetics (brand name: Lv Xun, product name: Su Xin vitality cream) for testing, and 3 polypeptides with moisturizing and spot lightening repair effects were screened, and finally one polypeptide with the best effect was determined for subsequent experiments. The polypeptide was obtained by sequencing, as shown in SEQ ID NO. 1.

[0034] In order to obtain sufficient amount of polypeptide for subsequent experiments, Hubei Qiangyao Biotechnology Co., Ltd. was entrusted to biosynthesize, and the purity was not less than 90%.

[0035] Example 1

[0036] This embodiment 1 provides a polypeptide composition containing moringa seed extract, and the preparation method is as follows: (1) Prepare physiological saline solution pre-cooled at 4°C, and respectively take 4 parts of moringa seed extract, 1 part of polypeptide derived from Potamogeton crispus and 2 parts of polypeptide derived from mulberry according to the proportion; (2) Put the physiological saline into a beaker, and add the moringa seed extract under ice bath condition while stirring, the stirring speed is 100 rpm, continue to stir for about 10 minutes after the addition is completed until completely mixed to obtain a mixed solution 1; (3) Continue to add the polypeptide derived from Potamogeton crispus and the polypeptide derived from mulberry to the mixed solution 1, and add them while stirring in the same way, the stirring speed is 80 rpm, continue to stir for about 10 minutes after the addition is completed until completely mixed to obtain a polypeptide composition.

[0037] Among them, the polypeptide derived from Potamogeton crispus is as shown in SEQ ID NO. 1, the polypeptide derived from mulberry is as shown in SEQ ID NO. 2, and the moringa seed extract is purchased from Gansu Yishengxiang Biotechnology Co., Ltd.

[0038] Embodiment 2

[0039] This embodiment 2 provides a polypeptide composition containing moringa seed extract, and the preparation method is as follows: (1) Prepare physiological saline solution pre-cooled at 4°C, and respectively take 8 parts of moringa seed extract, 1.5 parts of polypeptide derived from Potamogeton crispus and 3 parts of polypeptide derived from mulberry according to the proportion; (2) Put the physiological saline into a beaker, and add the moringa seed extract under ice bath condition while stirring, the stirring speed is 100 rpm, continue to stir for about 10 minutes after the addition is completed until completely mixed to obtain a mixed solution 1; (3) Continue to add the polypeptide derived from Potamogeton crispus and the polypeptide derived from mulberry to the mixed solution 1, and add them while stirring in the same way, the stirring speed is 80 rpm, continue to stir for about 10 minutes after the addition is completed until completely mixed to obtain a polypeptide composition.

[0040] Among them, the polypeptide derived from Potamogeton crispus is as shown in SEQ ID NO. 1, the polypeptide derived from mulberry is as shown in SEQ ID NO. 2, and the moringa seed extract is purchased from Gansu Yishengxiang Biotechnology Co., Ltd.

[0041] Embodiment 3

[0042] This embodiment 3 provides a polypeptide composition containing moringa seed extract, and the preparation method is as follows: (1) Prepare physiological saline solution pre-cooled at 4°C, and respectively take 11 parts of moringa seed extract, 3 parts of polypeptide derived from Potamogeton crispus and 4.5 parts of polypeptide derived from mulberry according to the proportion; (2) Put physiological saline into a beaker, under ice bath condition, add the extract of Moringa oleifera seed while stirring, the stirring speed is 100 rpm, continue to stir for about 10 min after the addition is completed until completely mixed to obtain a mixed solution 1; (3) Continue to add the polypeptide derived from Trapa and the polypeptide derived from Morus to the mixed solution 1, and add them while stirring at the same time, the stirring speed is 80 rpm, continue to stir for about 10 min after the addition is completed until completely mixed to obtain a polypeptide composition.

[0043] Among them, the polypeptide derived from Trapa is shown as SEQ ID NO. 1, the polypeptide derived from Morus is shown as SEQ ID NO. 2, and the extract of Moringa oleifera seed is purchased from Gansu Yishengxiang Biotechnology Co., Ltd.

[0044] Comparative Example 1

[0045] This comparative example applies the formula of Example 2, the difference is that no polypeptide derived from Trapa and no polypeptide derived from Morus are added.

[0046] Comparative Example 2

[0047] This comparative example applies the formula of Example 2, the difference is that no polypeptide derived from Morus is added.

[0048] Comparative Example 3

[0049] This comparative example applies the formula of Example 2, the difference is that no polypeptide derived from Trapa is added.

[0050] Comparative Example 4

[0051] This comparative example applies the formula of Example 2, the difference is that no extract of Moringa oleifera seed and no polypeptide derived from Morus are added.

[0052] Comparative Example 5

[0053] This comparative example applies the formula of Example 2, the difference is that no polypeptide derived from Trapa and no extract of Moringa oleifera seed are added.

[0054] Comparative Example 6

[0055] This comparative example applies the formula of Example 2, the difference is that no extract of Moringa oleifera seed is added.

[0056] Blank Control

[0057] The blank control is physiological saline, and no extract of Moringa oleifera seed, no polypeptide derived from Trapa and no polypeptide derived from Morus are added.

[0058] Experimental Example 1

[0059] The polypeptide compositions from Examples 1-3, Comparative Examples 1-6, and the blank control were used to formulate skin repair cosmetics. The formulation steps are as follows: The skin repair cosmetic formula includes the following components by weight, which should be weighed separately and mixed according to the specified proportions: 20 parts of polypeptide composition 8 parts of sodium hyaluronate 9 parts of cyclopentadimethylsiloxane 5 parts of PEG-10 polydimethylsiloxane 1 part of tocopherol acetate 1 part phenoxyethanol Finally, by adjusting the volume with solvent water, experimental test examples 1-3, comparative test examples 1-6, and blank test examples corresponding to the above Examples 1-3, Comparative Examples 1-6, and blank control were prepared.

[0060] Experimental Example 2

[0061] One hundred healthy women aged 30-50 years were recruited, with no history of skin allergies and mild to moderate facial dryness, fine lines, uneven skin tone, and pigmentation. These participants were randomly divided into 10 groups of 10 each. The skin-repairing cosmetics described in Experimental Test Examples 1-3, Control Test Examples 1-6, and the blank test examples were used on the entire face after cleansing, morning and evening, for 8 weeks. Skin hydration, melanin index, facial wrinkle area, and number of facial pigmentation spots were measured before use, at 4 weeks, and at 8 weeks to obtain the corresponding effects.

[0062] 1. Skin Moisture Content Detection: The Corneometer® CM825 was used to measure skin moisture content. During measurement, the instrument probe was gently and vertically pressed against the skin area to be tested, and the average moisture content value (unit: AU) was obtained. The average rate of change in moisture content was measured at the end of the experiment. The test results are shown in Table 1 below: Group Before use (AU) 4 weeks (AU) 8 weeks (AU) Change rate (%) Experimental test example 1 38.8 41.5 49.6 +27.8% Experimental test example 2 39.2 44.9 52.8 +34.7% Experimental test example 3 39.5 43.1 51.5 +30.4% Comparative test example 1 39.4 42.2 43.5 +10.4% Comparative test example 2 39.5 42.7 44.8 +13.4% Comparative test example 3 39.1 41.8 43.3 +10.7% Comparative test example 4 40.3 40.2 41.7 +3.5% Comparative test example 5 39.9 40.7 41.5 +4.0% Comparative test example 6 40.3 41.3 42.9 +6.5% Blank test example 40.2 40.5 41.2 +2.5%

[0063] As shown in the table above, after 8 weeks of use, the moisture content change rates of experimental test cases 1-3 were significantly better than those of the control test case and the blank test case. Among them, experimental test case 2 showed the best effect, with a change rate of nearly 35% after 8 weeks. The blank test case, on the other hand, showed almost no repair effect, with a change rate of only 2.5%, which may be due to interference from other ingredients in the cosmetic.

[0064] Regarding the comparative test cases, Comparative Test Case 1 showed only a 10.4% improvement, demonstrating that while Moringa seed extract has some moisturizing ability, it is limited. Comparative Test Cases 2 and 3 showed improvements of 13.4% and 10.7% respectively, indicating that the combination of a single peptide and Moringa seed extract has a moderate effect. Comparative Test Cases 4-6 were better than those lacking Moringa seed extract, showing the worst effect, almost approaching that of the test cases, indicating that using either peptide alone is insufficient to achieve a synergistic moisturizing effect.

[0065] 2. Skin melanin index detection: Mexameter® MX18 was used to measure and assess overall skin tone brightening. A lower value indicates a brighter skin tone and less melanin deposition. The test results are shown in Table 2 below: Group Before use (AU) 4 weeks (AU) 8 weeks (AU) Change rate (%) Experimental test example 1 160.5 153.3 142.6 -11.2% Experimental test example 2 158.8 146.3 133.2 -16.1% Experimental test example 3 161.0 148.8 136.1 -15.4% Comparative test example 1 158.3 154.4 152.2 -3.9% Comparative test example 2 161.2 153.1 151.4 -6.1% Comparative test example 3 158.6 154.2 146.1 -7.9% Comparative test example 4 160.3 158.7 156.2 -2.6% Comparative test example 5 161.5 157.4 157.0 -2.8% Comparative test example 6 159.1 154.6 149.2 -6.2% Blank test example 163.6 161.8 161.3 -1.4%

[0066] The data in the table above is analyzed as follows: All experimental test cases 1-3 showed significant and far superior skin brightening and spot-fading effects compared to the control group, with a specific MI value reduction of over 11.2%. Experimental test case 2, in particular, demonstrated the most outstanding effect, with a 16.1% reduction in melanin index after 8 weeks. This indicates that the formulation achieves the optimal balance of moisturizing, repairing, and spot-fading effects. The MI value of the blank test case remained almost unchanged, eliminating the interference of time or other external factors and proving the reliability of the experiment.

[0067] Comparative Test Example 1, containing only Moringa seed extract, showed a limited decrease in MI value, indicating that Moringa seeds themselves possess certain anti-inflammatory and antioxidant capabilities, and can slightly improve skin tone, but their effect is not significant when used alone. Comparative Test Example 2 was significantly weaker than Test Examples 1-3 with the complete formula. This demonstrates that mulberry peptides may play a key role in inhibiting tyrosinase activity and reducing melanin production; their absence leads to a significant reduction in the skin-brightening and blemish-fading effects. Comparative Test Example 3 was better than the group lacking mulberry peptides, but still far inferior to Test Examples 1-3 with the complete formula. This indicates that water shield peptides, by promoting barrier repair and the penetration of active ingredients, provide a better skin environment for mulberry peptides and Moringa seed extract to exert their effects, indirectly enhancing the overall brightening effect. Comparative Test Examples 4-6 showed the worst results, approaching the blank control group, further confirming that the three ingredients form an inseparable synergistic system.

[0068] 3. Facial wrinkle area detection: The VISIA skin analyzer was used to analyze the change rate (%) of wrinkle area in the crow's feet region around the eyes. The detection results are shown in Table 3 below: Group Before use baseline value 4 weeks test value 8 weeks test value Change rate (%) Experimental test example 1 12.53 10.82 10.03 -20.0% Experimental test example 2 12.37 11.63 9.31 -24.7% Experimental test example 3 12.57 11.24 9.84 21.7% Comparative test example 1 12.46 12.23 11.65 -6.5% Comparative test example 2 12.68 11.85 11.37 -10.3% Comparative test example 3 12.35 11.89 11.34 -8.2% Comparative test example 4 12.78 12.54 12.42 -2.8% Comparative test example 5 12.67 12.61 12.32 -2.8% Comparative test example 6 12.36 11.91 11.22 -9.2% Blank test example 12.74 12.76 12.60 -1.1%

[0069] The data in the table above is analyzed as follows: All experimental groups 1-3 with the complete formula exhibited remarkably significant anti-wrinkle effects, with wrinkle area reduced by more than 20%, far exceeding any control or blank test cases. In particular, experimental group 2 showed the peak effect, with wrinkle area reduced by 25.7% after 8 weeks. This indicates that, under this formulation, the moisturizing and anti-inflammatory effects of moringa seed, the repair and collagen-promoting effects of water shield peptides, and the antioxidant protection of mulberry peptides achieved an optimal synergistic balance. Meanwhile, the wrinkle area percentage in the blank test cases remained almost unchanged, further demonstrating the stability of the experimental environment and the reliability of the results.

[0070] Comparative Test 1, containing only Moringa seed extract, showed limited improvement in wrinkles, indicating that its anti-inflammatory and basic moisturizing capabilities provided only slight improvement. Comparative Test 3 was comparable to Test 1, but significantly weaker than the complete formula. This demonstrates that water shield peptides, as membrane protein-related peptides, play a central role in promoting skin cell migration, repair, and collagen synthesis, and are key to anti-wrinkle efficacy. Comparative Tests 2 and 6 were more effective than the group lacking water shield peptides, indicating that mulberry peptides reduce collagen damage through potent antioxidant activity, and Moringa seed extract provides a healthy environment for the skin through anti-inflammation; both indirectly support anti-wrinkle effects but cannot replace the direct repairing effect of water shield peptides. Comparative Tests 4 and 5 showed the worst results, approaching the control group, indicating that the absence of any two core ingredients would lead to the failure of the anti-wrinkle system.

[0071] Facial pigmentation: The number of pigmentation spots and the percentage change in total pigment area in the cheek area were analyzed using the VISIA skin analyzer. The results are shown in Tables 4 and 5 below. Group Before use baseline value 4 weeks test value 8 weeks test value Change rate (%) 31 28 21 Experimental test example 1 30 25 18 Experimental test example 2 31 26 20 Experimental test example 3 27 26 24 Comparative test example 1 27 25 22 Comparative test example 2 29 25 24 Comparative test example 3 28 27 25 Comparative test example 4 27 26 25 Comparative test example 5 27 25 22 Comparative test example 6 28 28 27

[0072] Table 5: Blank test example Group Before use baseline value (%) 8 weeks test value (%) Change rate (%) 5.7% 4.2% -26.3% Experimental test example 1 5.5% 3.7% -32.7% Experimental test example 2 5.8% 4.1% -29.3% Experimental test example 3 4.9% 4.5% -8.2% Comparative test example 1 5.2% 4.6% -11.5% Comparative test example 2 5.4% 4.8% -11.1% Comparative test example 3 5.0% 4.9% -2.0% Comparative test example 4 4.8% 4.7% -2.1% Comparative test example 5 5.1% 4.5% -11.8% Comparative test example 6 Blank test example 5.1% 5.0% -1.9%

[0073] Note: Percentage (%) of the area of ​​pigmentation in the total area analyzed.

[0074] The data in the table above is analyzed as follows: All experimental test cases demonstrated excellent results in reducing both the number and area of ​​pigmentation spots, with reduction rates 2-3 times higher than the control test cases. Test case 2 showed the most remarkable results, with a reduction of 12 spots and a total pigmentation area reduction of 32.7% after 8 weeks. This verifies that the formulation is the golden ratio for balancing "spot fading" and "repair / penetration enhancement" effects. Test case 3 showed roughly the same reduction in the number of spots as test case 2, but a slightly lower reduction rate in pigmentation area. This may be because the absorption and penetration efficiency of high-concentration peptides does not increase linearly, proving that higher concentrations are not always better; the optimal formulation is crucial. The blank test cases showed almost no change in the number and area of ​​spots, eliminating the interference of natural metabolism and demonstrating the effectiveness of the experiment.

[0075] The effects of comparative tests 2 and 6 were far inferior to those of the experimental test cases, but the difference from comparative test case 3 was not significant. This indicates that mulberry peptides may directly act on melanin metabolism, achieving the core efficacy of fading dark spots. Comparative test case 1 showed the worst effect, proving that the effect of moringa seed extract alone in fading dark spots is limited. The absence of any one peptide significantly reduced the effect, demonstrating that the three form a synergistic system. The effects of comparative tests 4 and 5 were almost identical to the blank test case, indicating that relying on only one peptide is insufficient to achieve the desired effect of fading dark spots.

[0076] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A polypeptide composition containing Moringa seed extract, characterized in that, Includes the following ingredients by weight: Moringa seed extract 3-12 parts 0.5-3.5 parts of polypeptides derived from water shield. 1-5 portions of polypeptides derived from mulberries The polypeptide derived from water shield is shown in SEQ ID NO.1, and the polypeptide derived from mulberry is shown in SEQ ID NO.

2.

2. The polypeptide composition containing Moringa seed extract as described in claim 1, characterized in that, Includes the following ingredients by weight: Moringa seed extract 8 parts 1.5 portions of polypeptides derived from water shield. Three portions of polypeptides derived from mulberries.

3. The polypeptide composition containing Moringa seed extract as described in claim 1 or 2, characterized in that, in, The extraction method for the polypeptide SEQ ID NO.1 is as follows: (1) Raw material pretreatment: Wash the water shield sample with pre-cooled distilled water to remove impurities and absorb the moisture with filter paper; (2) Grinding: The water shield sample was placed in liquid nitrogen and frozen quickly. It was then transferred to a pre-cooled mortar and pestle and ground into an extremely fine powder. (3) Crude extraction: Transfer the ground powder to a beaker containing pre-cooled Tris-HCl extraction buffer, stir continuously on ice for 30-60 min, centrifuge at 3000 rpm for 10-15 min at 4℃, and collect the supernatant; centrifuge at 20000 rpm for 30-45 min at 4℃, and collect the supernatant; centrifuge at 100000 rpm for 30-45 min at 4℃, and collect the precipitate; (4) Fine purification: The above precipitate was redissolved in Tris-HCl buffer containing 1% DDM, and slowly shaken at 4°C for 8-12 h. The mixture after incubation was then centrifuged at 100,000 rpm at 4°C for 60-90 min to obtain the supernatant. (5) Purification: The protein peaks were collected by sequentially passing the affinity chromatography column and the cation exchange chromatography column and then purified by dialysis. The peptides corresponding to the multiple protein peaks were added to the cosmetics of our company for testing. Three peptides were screened and found to have moisturizing, spot-fading and repairing effects. Finally, the peptide with the best effect was determined for subsequent experiments. This peptide is shown in SEQ ID NO.

1.

4. The polypeptide composition containing Moringa seed extract as described in claim 1 or 2, characterized in that, in, The extraction method for the polypeptide SEQ ID NO.2 is as follows: (1) Raw material pretreatment: Wash the mulberry samples with pre-cooled distilled water to remove impurities and absorb the moisture with filter paper; (2) Grinding: The mulberry sample was placed in liquid nitrogen and frozen quickly. It was then transferred to a pre-cooled mortar and pestle and ground into an extremely fine powder. (3) Extraction: Transfer the ground powder to a beaker containing pre-cooled Tris-HCl extraction buffer, stir continuously on ice for 30-60 min, centrifuge at 8000 rpm for 30-45 min at 4℃, and collect the supernatant to obtain mulberry extract; (4) Purification: The protein peaks were sequentially chromatographically passed through an affinity chromatography column and a cation exchange chromatography column. Multiple protein peaks were collected and purified by dialysis. The peptides corresponding to the multiple protein peaks were added to the cosmetics of our company for testing. Five peptides were screened to have moisturizing, spot-fading and repairing effects. Finally, the peptide with the best effect was determined for subsequent experiments. This peptide is shown in SEQ ID NO.

2.

5. The use of the polypeptide composition containing Moringa seed extract as described in any one of claims 1-4 in the preparation of skin repair drugs or cosmetics.

6. The application as described in claim 5, characterized in that, The skin repair cosmetic formula includes the following components in parts by weight: 10-25 parts of polypeptide composition 5-10 parts moisturizer 7-15 parts moisturizer 2-5 parts emulsifier Antioxidant 0.5-1 part Preservative 0.5-1.5 parts Adjust the volume using water as a solvent.

7. The application as described in claim 6, characterized in that, in, The emollient is one or both of cyclopentamethoxysiloxane and squalane; the emulsifier is PEG-10 polydimethylsiloxane.

8. The application as described in claim 6, characterized in that, The moisturizer is one or both of 1,3-propanediol and sodium hyaluronate.

9. The application as described in claim 6, characterized in that, The antioxidant is one or more of tocopheryl acetate, vitamin C, and vitamin E.

10. The application as described in claim 6, characterized in that, The preservative is one or more of hexanediol, phenoxyethanol, and chlorphenesin.