Collagen peptide composition for improving skin elasticity and gloss and production process thereof

Abstract

The invention provides a collagen peptide composition. The collagen peptide composition comprises collagen tripeptide and mulberry exosome. The invention also provides a method for preparing the collagen peptide composition.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of cosmetic skin care, in particular to orally consumable collagen peptide compositions comprising collagen peptides. The present application also relates to the preparation process thereof. BACKGROUND

[0002] Collagen is a protein that is abundant in the human body, widely exists in the skin, tendon, ligament, cartilage and blood vessels, and plays a role in supporting, protecting, connecting and maintaining elasticity. Collagen and its degradation product collagen peptides are widely used in cosmetics for improving skin quality. Collagen tripeptide (CTP) is a kind of collagen peptide with smaller molecular weight and simpler structure. It is composed of three amino acids (mainly glycine-proline-hydroxyproline), and is considered to be a form of collagen peptide with higher absorption efficiency. Exosomes, as small vesicles containing bioactive substances produced by cells, have functions of regulating cell function and promoting tissue repair, and have attracted attention in the field of cosmetic skin care, but there is no report on the promotion of collagen tripeptide efficacy by plant exosomes. SUMMARY

[0003] In one aspect, provided herein is a method of preparing a collagen peptide composition, comprising mixing: collagen tripeptide, mulberry exosome suspension, and water, wherein the mulberry exosome suspension is prepared by: 1) filtering mulberry homogenate, collecting the filtrate; centrifuging the filtrate at 800g for 10 minutes at 4°C, collecting the supernatant; centrifuging at 10,000g for 30 minutes, collecting the supernatant; centrifuging at 150,000g for 50 minutes, collecting the precipitate; resuspending the precipitate with PBS buffer; or 2) filtering mulberry homogenate, collecting the filtrate; centrifuging the filtrate at 800g for 10 minutes at 4°C, collecting the supernatant; centrifuging at 10,000g for 30 minutes, collecting the supernatant; centrifuging at 100,000g for 20 minutes, collecting the supernatant; centrifuging at 150,000g for 50 minutes, collecting the precipitate; resuspending the precipitate with PBS buffer.

[0004] In some embodiments, the method comprises mixing: collagen tripeptide, mulberry exosome suspension, hydroxyproline, and water.

[0005] In some embodiments, the method comprises mixing: collagen tripeptide, mulberry exosome suspension, hydroxyproline, sweetener, soluble dietary fiber, and water.

[0006] In some embodiments, the method comprises mixing the following components: collagen tripeptide, mulberry exosome suspension, hydroxyproline, sweetener, soluble dietary fiber, pinenut powder, and water.

[0007] In some embodiments, the sweetener is selected from the group consisting of sorbitol, sucralose, and combinations thereof.

[0008] In some embodiments, the dietary fiber is inulin.

[0009] In some embodiments, the method comprises mixing the following components in the following weight parts: 5-10 parts of bovine collagen tripeptide, 5-15 parts of sorbitol, 5-20 parts of soluble inulin, 2-8 parts of the mulberry exosome suspension, 2-5 parts of hydroxyproline, 0.01-0.05 parts of sucralose, 1-5 parts of pinenut powder, and 40-100 parts of water.

[0010] In some embodiments, the method comprises mixing the following components in the following weight parts: 8 parts of bovine collagen tripeptide, 10 parts of sorbitol, 10 parts of soluble inulin, 3 parts of the mulberry exosome suspension, 3 parts of hydroxyproline, 0.01 parts of sucralose, 2 parts of pinenut powder, and 60 parts of water, followed by steam sterilization at 121°C for 20 minutes.

[0011] In some embodiments, the mulberry exosome suspension is 8 mg / L in protein content.

[0012] In some embodiments, the exosome particle size in the mulberry exosome suspension is less than 80 nm.

[0013] In another aspect, provided herein is a collagen peptide composition prepared by the above method.

[0014] The collagen peptide composition provided herein has the effect of improving skin elasticity and luster after taking. DETAILED DESCRIPTION

[0015] Unless otherwise defined, all technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art.

[0016] In this document, the terms "comprising" and "including" generally mean the inclusion of the specified elements but not to the exclusion of other elements. In addition, the terms "comprising" and "including" are also intended to encompass the case where one or more of the specified elements are present.

[0017] Unless otherwise specified, the proportions in parts used herein are weight proportions.

[0018] In some embodiments, the present application provides a drink containing collagen tripeptide (or collagen peptide composition) comprising: collagen tripeptide, mulberry exosome, sweetener, and water.

[0019] In more specific embodiments, the drink containing collagen tripeptide (or collagen peptide composition) provided by the present application comprises: collagen tripeptide, mulberry exosome, sweetener, dietary fiber, and water.

[0020] In more specific embodiments, the drink containing collagen tripeptide (or collagen peptide composition) provided by the present application comprises: collagen tripeptide, mulberry exosome, sweetener, dietary fiber, sweetener, and water.

[0021] In more specific embodiments, the drink containing collagen tripeptide (or collagen peptide composition) provided by the present application comprises: collagen tripeptide, mulberry exosome, sweetener, dietary fiber, hydroxyproline, sweetener, and water.

[0022] Preferably, the sweetener is selected from the group consisting of sorbitol, sucralose, and combinations thereof.

[0023] Preferably, the dietary fiber is soluble, such as soluble inulin.

[0024] Preferably, the ratio of collagen tripeptide to mulberry exosome suspension (protein concentration 8 mg / L) in the drink is 5:1 to 2:1.

[0025] Preferably, the ratio of collagen tripeptide to soluble inulin in the drink is 1:1 to 1:4.

[0026] Preferably, the particle size of the mulberry exosome is less than 80 nm.

[0027] More preferably, the drink comprises: 5-10 parts of bovine collagen tripeptide, 5-15 parts of sorbitol, 5-20 parts of soluble inulin, 2-8 parts of mulberry exosome suspension (protein concentration 8 mg / L), 2-5 parts of hydroxyproline, 0.01-0.05 parts of sucralose, 1-5 parts of pinus roxburghii powder, and 40-100 parts of water.

[0028] In some embodiments, the present application provides a centrifugation method for preparing mulberry exosomes with a particle size of less than 80 nm, comprising: centrifuging the mulberry homogenate solution at 800g for 10 minutes at 4°C to collect the supernatant; centrifuging at 10,000g for 30 minutes to collect the supernatant; centrifuging at 100,000g for 20 minutes to collect the supernatant; centrifuging at 150,000g for 50 minutes to collect the precipitate, and resuspending the precipitate with PBS buffer.

[0029] The present inventors have found that the inclusion of mulberry exosomes in a drink containing collagen tripeptide helps to improve the elasticity and gloss of the skin of the subject, and the effect is faster.

[0030] The present application is described in detail below through specific examples.

[0031] Example 1 Preparation of mulberry exosomes Fresh mulberries (black purple) were taken and the surface was cleaned with clean water, then washed with deionized water for 2 times. The washed mulberries were added into a homogenizer, and pre-cooled PBS buffer was added at a volume ratio of 1:5, and homogenized (20000 rpm) for 30 seconds. The obtained homogenate was filtered with a 100 mesh screen, and the filtrate was collected. The filtrate was centrifuged at 800g for 10 minutes at 4°C, and the supernatant was collected; centrifuged at 10,000g for 30 minutes, and the supernatant was collected; centrifuged at 150,000g for 50 minutes, and the precipitate was collected. The precipitate was resuspended with PBS buffer.

[0032] The size of the exosomes was observed by scanning electron microscopy, the protein content was determined by the Bradford method, and the concentration of the exosome suspension prepared in different batches was determined by the protein content. The obtained exosomes had a diameter of 50 to 160 nm, and the protein content was 25-35 mg / L. The exosome concentration was diluted to 8 mg / L with PBS buffer, and stored at 4°C for standby.

[0033] Example 2 Optimization of mulberry exosome preparation The exosome suspension was prepared according to the method of Example 1, except that the centrifugation process was different from Example 1. The centrifugation process was as follows: the filtrate was centrifuged at 800g for 10 minutes at 4°C, and the supernatant was collected; centrifuged at 10,000g for 30 minutes, and the supernatant was collected; centrifuged at 100,000g for 20 minutes, and the supernatant was collected; centrifuged at 150,000g for 50 minutes, and the precipitate was collected. The precipitate was resuspended with PBS buffer.

[0034] Here, two-step ultracentrifugation (100,000g and 150,000g) was used to obtain exosomes with smaller particle size. The obtained exosomes had a diameter of 30 to 80 nm, as observed by scanning electron microscopy. The measured protein content was 30-38 mg / L. The exosome concentration was diluted to 8 mg / L with PBS buffer, and stored at 4°C for standby.

[0035] Example 3 Preparation of collagen peptide composition The following components were mixed in a weight ratio of 8 parts of bovine collagen tripeptide, 10 parts of sorbitol, 10 parts of soluble inulin, 3 parts of exosome suspension prepared in Example 1 (concentration 8 mg / L), 3 parts of hydroxyproline, 0.01 parts of sucralose, 2 parts of needle cherry powder, and 60 parts of water. Steam sterilization at 121°C for 20 minutes, and after cooling, sterilely packaged into sterile bags, 50 mL per bag.

[0036] Example 4 Preparation of collagen peptide composition Prepared by the method of Example 3, except that the exosome suspension used was the exosome suspension prepared in Example 2 (concentration 8 mg / L).

[0037] Example 5 (comparative example) Collagen peptide composition preparation Prepared by the method of Example 3, except that no exosome suspension was added.

[0038] Example 5 Skin elasticity and glossiness detection Subject inclusion criteria: female aged 25-55 years; dry skin; self-perceived dull, slack, lack of gloss and elasticity on the face; no obvious skin lesions and scars on the facial skin; able to maintain a regular life during the experiment.

[0039] Subject exclusion criteria: suffering from infectious skin diseases or atopic dermatitis; receiving drug treatment; in the pregnancy or lactation period; not agreeing to sign the informed consent form.

[0040] A total of 60 people were enrolled and randomly divided into 3 groups, each group of 20 people, including: Example 3 group, taking one bag of collagen peptide composition prepared in Example 3 per day; Example 4 group, taking one bag of collagen peptide composition prepared in Example 4 per day; Example 5 group, taking one bag of collagen peptide composition prepared in Example 5 per day.

[0041] The total facial skin elasticity R2 value (taking the average of three detections) was detected by Cutometer® dual MPA580 (Germany CK) and the skin glossiness was detected by Glossymeter® GL200 (Germany CK) (taking the average of three detections) on the experimental start day (0 days), 28 days and 56 days.

[0042] The skin elasticity detection results are shown in Table 1. The R2 value is between 0 and 1, and a higher R2 value reflects better skin elasticity.

[0043] Table 1 Skin elasticity detection results

[0044] Different lowercase letters in the same column indicate that the detection values at different times are significantly different (p<0.05), and the same lowercase letters indicate that there is no significant difference (p>0.05); different capital letters in the same column indicate that the detection values between groups are significantly different (p<0.05), and the same capital letters indicate that there is no significant difference (p>0.05). p p p p

[0045] ​​​​The results of skin gloss measurement (incident light angle 60°) are shown in Table 2. Larger measurements indicate better skin gloss.

[0046] Table 2 Results of Skin Glossiness Test

[0047] Different lowercase letters on the same row's shoulder label indicate significant differences in test values ​​at different times. p <0.05), with the same lowercase letter indicating no significant difference ( p >0.05); different capital letters on the same column indicate significant differences in test values ​​between groups. p <0.05, with the same uppercase letter indicating no significant difference ( p >0.05).

[0048] As can be seen from the results in Tables 1 and 2, the collagen peptide composition containing collagen tripeptides prepared in Example 5 showed a certain effect in improving skin elasticity and radiance at 56 days. However, in terms of demonstrating this improvement effect earlier and in terms of final test values, the collagen peptide compositions prepared in Examples 3 and 4 were more effective, showing significant improvement in skin elasticity and radiance at 28 days. Comparing the results of the Example 3 and Example 4 groups, it can be found that exosomes with smaller particle sizes are superior to those with relatively larger particle sizes. This is reflected in the significant improvement in skin elasticity at 56 days compared to 28 days (see the results of Example 4 group in Table 1), and the better skin elasticity and radiance at 56 days (see the results of Example 3 and Example 4 groups at 56 days in Tables 1 and 2).

Claims

1. A method for preparing a collagen peptide composition, comprising mixing the following components: collagen tripeptide, mulberry exosome suspension, and water. The mulberry exosome suspension is prepared by any of the following processes: 1) After filtering the mulberry homogenate, collect the filtrate; centrifuge the filtrate at 800g for 10 minutes at 4°C and collect the supernatant; centrifuge at 10,000g for 30 minutes and collect the supernatant; centrifuge at 150,000g for 50 minutes and collect the precipitate; resuspend the precipitate in PBS buffer; or 2) After filtering the mulberry homogenate, collect the filtrate; centrifuge the filtrate at 800g for 10 minutes at 4°C and collect the supernatant; centrifuge at 10,000g for 30 minutes and collect the supernatant; centrifuge at 100,000g for 20 minutes and collect the supernatant; centrifuge at 150,000g for 50 minutes and collect the precipitate; resuspend the precipitate in PBS buffer.

2. The method of claim 1, comprising mixing the following components: collagen tripeptide, mulberry exosome suspension, hydroxyproline and water.

3. The method of claim 2, comprising mixing the following components: collagen tripeptide, mulberry exosome suspension, hydroxyproline, sweetener, soluble dietary fiber, acerola cherry powder, and water.

4. The method of claim 3, wherein the sweetener is selected from sorbitol, sucralose, and combinations thereof.

5. The method of claim 3, wherein the dietary fiber is inulin.

6. The method of claim 1, comprising mixing the following components in parts by weight: 5-10 parts bovine collagen tripeptide, 5-15 parts sorbitol, 5-20 parts soluble inulin, 2-8 parts mulberry exosome suspension, 2-5 parts hydroxyproline, 0.01-0.05 parts sucralose, 1-5 parts acerola cherry powder, and 40-100 parts water.

7. The method of claim 6, comprising mixing the following components in parts by weight: 8 parts bovine collagen tripeptide, 10 parts sorbitol, 10 parts soluble inulin, 3 parts the mulberry exosome suspension, 3 parts hydroxyproline, 0.01 parts sucralose, 2 parts acerola cherry powder and 60 parts water, and then steam sterilizing at 121°C for 20 minutes.

8. The method of claim 1, wherein the mulberry exosome suspension has a protein content of 8 mg / L.

9. The method of claim 1, wherein the exosome particle size in the mulberry exosome suspension is less than 80 nm.

10. The collagen peptide composition prepared by the method of any one of claims 1-9.

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