A wrinkle-lightening and firming triple-active collagen composition and its preparation method and application

Abstract

The present invention discloses a wrinkle-lightening and firming triple active collagen composition and its preparation method and application, and relates to the technical field of cosmetics. The composition includes: recombinant humanized collagen type I, recombinant humanized collagen type III, recombinant humanized collagen type XVII, ectoine, ergothioneine, nicotinamide, sea fennel water extract, Centella asiatica extract, Antrodia camphorata extract, Hippophae rhamnoides extract, Polygonum multiflorum root extract, Dendrobium candidum extract, retinol and erythritol; at the same time, it can also include three collagen derivatives. The composition prepared by the present invention by reasonably matching the raw materials can not only effectively promote cell proliferation and regeneration and cell migration, but also can significantly improve skin elasticity, firmness and lighten fine lines.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and more particularly to a wrinkle-lightening and firming triple-active collagen composition and a preparation method and application thereof. Background Art

[0002] Collagen is an important protein in the human body, accounting for more than 30% of the total protein content in the human body. An adult contains about 3 kilograms of collagen. It is the most important raw material for the human body and can be called the scaffold of human life. It is divided into 28 types according to the amino acid sequence and is widely distributed in human tissues, including skin mucosa, muscles, hair, bones, cartilage, joints, organs and other tissues. Among them, type I, II, and III collagen account for more than 90% of the total collagen. Its main function is to maintain the morphology and structure of skin and tissues and organs. It is also an important raw material for repairing damaged tissues.

[0003] Skin aging is a complex biological behavior involving multiple skin components, including natural aging and extrinsic aging. Although natural aging and extrinsic aging are completely different in terms of biology, biochemistry and molecular mechanisms, they both lead to significant changes in elastic fibers. Intrinsic aging is manifested by a decrease in the number of elastic fibers, and photoaging is manifested by the accumulation of elastic fiber substances. With the significant increase in age and sun exposure, the content of elastin will decrease. Studies have shown that new elastin is not produced after adulthood. One of the focuses of skin anti-aging or beauty is to increase the amount of elastin in adults, but based on this principle, there are few products that can achieve significant improvements in skin quality.

[0004] With the development of recombinant technology, recombinant humanized collagen such as type I and type III have been gradually developed, but the current research is still relatively simple and not in-depth enough; more importantly, whether there is a complex synergy between collagen and collagen or other active ingredients, whether there are related synergistic functions, and whether it can enhance the wrinkle reduction, firming and anti-aging effects achieved when used alone. The industry currently lacks the corresponding mechanism and data. Summary of the invention

[0005] In view of this, the present invention provides a wrinkle-lightening and firming triple-active collagen composition, and a preparation method and application thereof.

[0006] In order to achieve the above object, the present invention adopts the following technical solution:

[0007] A wrinkle-lightening and firming triple-active collagen composition, wherein each 100 mL of the composition comprises:

[0008] 0.5-2 mg of recombinant humanized collagen type I and / or a protein having the same function as the recombinant humanized collagen type I, obtained by substitution and / or deletion and / or addition of one or several amino acid residues, 2.5-10 mg of recombinant humanized collagen type III and / or a protein having the same function as the recombinant humanized collagen type III, obtained by substitution and / or deletion and / or addition of one or several amino acid residues, and 0.5-2 mg of recombinant humanized collagen type XVII and / or a protein having the same function as the recombinant humanized collagen type XVII, obtained by substitution and / or deletion and / or addition of one or several amino acid residues;

[0009] Preferably, the composition is used for wrinkle reduction and skin tightening.

[0010] Preferably, each 100 mL of the composition further comprises 0.25-0.8 mg of ectoine, 0.12-0.46 mg of ergothioneine, 0.32-0.51 mg of nicotinamide and 0.15-0.40 mg of sea fennel water extract.

[0011] Preferably, each 100 mL of the composition further comprises 0.10-0.37 mg of Centella asiatica extract, 0.12-0.28 mg of Camphorata extract, 0.08-0.17 mg of Hippophae rhamnoides extract, 0.05-0.15 mg of Polygonum multiflorum root extract, 0.01-0.10 mg of Dendrobium officinale extract, 0.01-0.15 mg of retinol, and 0.01-0.08 mg of erythritol.

[0012] Preferably, the preparation method of the sea fennel water extract is: crushing the sea fennel, passing through a 40-mesh sieve, adding the sea fennel powder to distilled water in a ratio of sea fennel: distilled water m:v=1:8, stirring evenly, extracting by water bath ultrasonication, filtering, centrifuging, concentrating and drying the supernatant, and then obtaining the sea fennel extract;

[0013] Preferably, the water bath temperature is 50°C, the ultrasonic power is 500W, and the extraction time is 2h;

[0014] Preferably, the centrifugation is performed at 800 rpm for 5 min.

[0015] Preferably, the Antrodia camphorata extract is a water extract;

[0016] Preferably, the preparation method of the water extract is: crushing Antrodia camphorata, passing through a 0.42 mm sieve, adding Antrodia camphorata powder to distilled water at a ratio of Antrodia camphorata: distilled water m:v=1:10, stirring evenly, extracting by heating in a water bath, filtering, centrifuging, concentrating and drying the supernatant to obtain Antrodia camphorata extract;

[0017] Preferably, the water bath temperature is 35°C and the extraction is performed for 8 hours;

[0018] Preferably, the centrifugation is performed at 500 rpm for 10 min.

[0019] As the same inventive concept as the above technical solution, the present invention also claims a method for preparing the above composition, comprising the following steps:

[0020] (1) mixing recombinant humanized type I collagen, recombinant humanized type III collagen, and recombinant humanized type XVII collagen to obtain multiple humanized collagens;

[0021] (2) mixing ectoine, ergothioneine, sea fennel, Centella asiatica and water, heating and stirring to dissolve evenly, and keeping the temperature to obtain a mixed solution A;

[0022] (3) After the mixed solution A cools down, niacinamide, retinol and erythritol are added in sequence, and heated and stirred until completely dissolved to obtain a mixed solution B;

[0023] (4) After the mixed solution B cools down, add multiple humanized collagen, Antrodia camphorata extract, Hippophae rhamnoides extract, Polygonum multiflorum root extract and Dendrobium officinale extract in sequence, stir evenly and homogenize to obtain a wrinkle-lightening and firming triple active collagen composition.

[0024] Preferably, in step (2), the heating temperature is 65-75°C, the insulation time is 15-30 min, the stirring speed is 300-400 rpm, and the stirring time is 5-10 min; in step (3), the cooling is 50-60°C, the heating time is 15-20 min, the stirring speed is 550-650 rpm, and the stirring time is 3-8 min.

[0025] Preferably, in step (4), the cooling speed is 35-40°C, the stirring speed is 400-500 rpm, and the stirring time is 5-15 min; the homogenization speed is 2100-2800 rpm, and the homogenization time is 8-12 min.

[0026] As the same inventive concept as the above technical solution, the present invention also claims to protect the use of the above composition or the composition prepared by the above method, which is the use in preparing wrinkle-lightening and firming skin care products, cosmetics or medicines;

[0027] Preferably, the composition improves cell viability and promotes cell proliferation;

[0028] Preferably, the composition promotes skin firmness and elasticity.

[0029] As an inventive concept identical to the above technical solution, the present invention also seeks to protect a product having wrinkle-lightening and firming effects, wherein the product comprises the above wrinkle-lightening and firming triple active collagen composition.

[0030] It can be seen from the above technical solutions that, compared with the prior art, the present invention discloses a triple active collagen composition for lightening and firming wrinkles, and a preparation method and application thereof, with the following beneficial effects:

[0031] The invention scientifically and reasonably combines recombinant humanized type I collagen, recombinant type III collagen, recombinant type XVII collagen, ectoine, ergothioneine, nicotinamide and a variety of plant extracts, which can effectively promote cell proliferation and regeneration and cell migration, and can significantly improve skin elasticity and firmness and reduce fine lines; wherein, recombinant humanized type I collagen has important functional sites of human type I collagen, has good biocompatibility and high biological activity, and can significantly promote the proliferation, adhesion and migration of human skin fibroblasts; recombinant humanized type III collagen itself has good moisturizing effect, can enhance cell metabolism, promote keratinocyte differentiation and skin renewal, plays a promoting role in maintaining normal skin function, and has tissue damage repair ability; recombinant type XVII humanized collagen is an important component of epidermal stem cell hemidesmosomes in the human body, plays an important role in cell aging and skin differentiation, and also has the characteristics of high water solubility and high biological activity. DETAILED DESCRIPTION

[0032] The following will be described clearly and completely in conjunction with the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0033] Ectoine, ergothioneine, niacinamide and natural active ingredients extracted from different plants all have good antioxidant activity, repair and whitening effects. They can prevent free radical damage to the skin, reduce the production of melanin, and promote cell regeneration. At the same time, they can also fight bacteria and inflammation, reduce the growth of bacteria on the skin, and play a role in protecting and repairing the skin.

[0034] The instruments, reagents, materials, experimental animals, etc. used in the present invention can be obtained through conventional commercial means.

[0035] The embodiment involves three kinds of recombinant humanized collagen, wherein the amino acid sequence of recombinant type I humanized collagen is:

[0036] GEKGSPGADGPAGAPGTPGPQGIAGQRGVVGLPGQRGERGFPGLPGPSGEPGKQGPSGASGEKGSPGADGPAGAPGTPGPQGIAGQRGVVGLPGQRGERGFPGLPGPSGEPGKQGPSGASGE KGSPGADGPAGAPGTPGPQGIAGQRGVVGLPGQRGERGFPGLPGPSGEPGKQGPSGASGEKGSPGADGPAGAPGTPGPQGIAGQRGVVGLPGQRGERGFPGLPGPSGEPGKQGPSGAS, such as SEQ Shown as ID NO.1.

[0037] The amino acid sequence of recombinant humanized type III collagen is:

[0038] GERGAPGFRGPAGPNGIPGEKGPAGERGAPGEROAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGFNGIPGEKGPAGERGAPGE RGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAP, such as SEQ Shown as ID NO.2.

[0039] The amino acid sequence of recombinant humanized type XVII collagen is:

[0040] GADFAGDLDYNELAVRVSESMQRQGLLQGMAYTVQGPPGQPGPQGPPGISKVFSAYSNVTADLMDFFQTYGAIQGPPGQKGEMGTPPGPKGDRGPAGPPGHPGPPGPRGHKGEKGDKGDQ, as shown in SEQ ID NO.3.

[0041] Example 1

[0042] A wrinkle-lightening and firming triple-active collagen composition, wherein each 100 mL of the composition comprises:

[0043] 0.5mg recombinant humanized type I collagen, 5mg recombinant humanized type III collagen, 1.5mg recombinant humanized type XVII collagen, 0.25mg ectoine, 0.24mg ergothioneine, 0.44mg niacinamide and 0.40mg sea fennel water extract, 0.26mg Centella asiatica extract, 0.17mg Antrodia cinnamomea extract, 0.08mg Hippophae rhamnoides extract, 0.08mg Polygonum multiflorum root extract, 0.07mg Dendrobium officinale extract, 0.15mg retinol, 0.05mg erythritol.

[0044] The preparation method is as follows:

[0045] (1) Preparation of sea fennel water extract: crush sea fennel, pass through a 40-mesh sieve, add sea fennel powder into distilled water in a ratio of sea fennel: distilled water (m:v=1:8), stir evenly, extract by water bath ultrasonication at 50°C and 800W for 2h, filter, centrifuge at 800rpm for 5min, concentrate the supernatant, and dry to obtain the sea fennel water extract;

[0046] (2) Preparation of Antrodia camphorata aqueous extract: Grind Antrodia camphorata and pass through a 0.42 mm sieve. Add Antrodia camphorata powder into distilled water at a ratio of 1:10, stir evenly, heat in a water bath at 35°C for 8 h, filter, centrifuge at 500 rpm for 10 min, concentrate and dry the supernatant to obtain Antrodia camphorata aqueous extract;

[0047] (3) mixing recombinant humanized type I collagen, recombinant humanized type III collagen, and recombinant humanized type XVII collagen to obtain multiple humanized collagens;

[0048] (4) mixing ectoine, ergothioneine, sea fennel, Centella asiatica and water, heating and stirring to dissolve at 65° C. and 400 rpm for 8 min, and keeping the temperature for 24 min to obtain a mixed solution A;

[0049] (5) After the mixed solution A is cooled to 50°C, niacinamide, retinol and erythritol are added in sequence, heated for 17 minutes, and stirred at 650 rpm for 5 minutes to completely dissolve them, thereby obtaining a mixed solution B;

[0050] (6) After the mixed solution B is cooled to 35°C, multiple humanized collagen, Antrodia camphorata extract, Hippophae rhamnoides extract, Polygonum multiflorum root extract and Dendrobium officinale extract are added in sequence, stirred at 450 rpm for 15 min, and homogenized at 2400 rpm for 8 min to obtain a wrinkle-lightening and firming triple active collagen composition.

[0051] Example 2

[0052] A wrinkle-lightening and firming triple-active collagen composition, wherein each 100 mL of the composition comprises:

[0053] 1mg recombinant humanized collagen type I, 7.5mg recombinant humanized collagen type III, 2mg recombinant humanized collagen type XVII, 0.60mg ectoine, 0.12mg ergothioneine, 0.38mg niacinamide and 0.31mg sea fennel water extract, 0.37mg Centella asiatica extract, 0.23mg Antrodia cinnamomea extract, 0.11mg sea buckthorn extract, 0.05mg Polygonum multiflorum root extract, 0.04mg Dendrobium officinale extract, 0.10mg retinol, 0.08mg erythritol;

[0054] The preparation method is as follows:

[0055] (1)-(2) Same as Example 1;

[0056] (3) mixing recombinant humanized type I collagen, recombinant humanized type III collagen, and recombinant humanized type XVII collagen to obtain multiple humanized collagens;

[0057] (4) mixing ectoine, ergothioneine, sea fennel, Centella asiatica and water, heating and stirring to dissolve at 60° C. and 350 rpm for 5 min, and keeping the temperature for 30 min to obtain a mixed solution A;

[0058] (5) After the mixed solution A is cooled to 55°C, niacinamide, retinol and erythritol are added in sequence, heated for 20 minutes, and stirred at 600 rpm for 3 minutes to completely dissolve them, thereby obtaining a mixed solution B;

[0059] (6) After the mixed solution B is cooled to 38°C, multiple humanized collagen, Antrodia camphorata extract, Hippophae rhamnoides extract, Polygonum multiflorum root extract and Dendrobium officinale extract are added in sequence, stirred at 500 rpm for 5 min, and homogenized at 2800 rpm for 10 min to obtain a wrinkle-lightening and firming triple active collagen composition.

[0060] Example 3

[0061] A wrinkle-lightening and firming triple-active collagen composition, wherein each 100 mL of the composition comprises:

[0062] 1.5mg recombinant humanized collagen type I, 10mg recombinant humanized collagen type III, 0.5mg recombinant humanized collagen type XVII, 0.45mg ectoine, 0.36mg ergothioneine, 0.51mg niacinamide and 0.15mg sea fennel water extract, 0.18mg Centella asiatica extract, 0.28mg Antrodia cinnamomea extract, 0.14mg Hippophae rhamnoides extract, 0.15mg Polygonum multiflorum root extract, 0.01mg Dendrobium officinale extract, 0.06mg retinol, 0.01mg erythritol;

[0063] The preparation method is as follows:

[0064] (1)-(2) Same as Example 1;

[0065] (3) mixing recombinant humanized type I collagen, recombinant humanized type III collagen, and recombinant humanized type XVII collagen to obtain multiple humanized collagens;

[0066] (4) mixing ectoine, ergothioneine, sea fennel, Centella asiatica and water, heating and stirring to dissolve at 75° C. and 300 rpm for 10 min, and keeping the temperature for 15 min to obtain a mixed solution A;

[0067] (5) After the mixed solution A is cooled to 60°C, niacinamide, retinol and erythritol are added in sequence, heated for 15 minutes, and stirred at 550 rpm for 8 minutes to completely dissolve them, thereby obtaining a mixed solution B;

[0068] (6) After the mixed solution B is cooled to 40°C, multiple humanized collagen, Antrodia camphorata extract, Hippophae rhamnoides extract, Polygonum multiflorum root extract and Dendrobium officinale extract are added in sequence, stirred at 400 rpm for 10 min, and homogenized at 2100 rpm for 12 min to obtain a wrinkle-lightening and firming triple active collagen composition.

[0069] Example 4

[0070] A wrinkle-lightening and firming triple-active collagen composition, wherein each 100 mL of the composition comprises:

[0071] 2mg recombinant humanized collagen type I, 2.5mg recombinant humanized collagen type III, 1mg recombinant humanized collagen type XVII, 0.80mg ectoine, 0.46mg ergothioneine, 0.32mg niacinamide and 0.23mg sea fennel water extract, 0.10mg Centella asiatica extract, 0.12mg Antrodia cinnamomea extract, 0.17mg Hippophae rhamnoides extract, 0.11mg Polygonum multiflorum root extract, 0.10mg Dendrobium officinale extract, 0.01mg retinol, 0.03mg erythritol;

[0072] The preparation method is the same as that in Example 2.

[0073] Comparative Example

[0074] Group A: The difference from Example 2 is that the recombinant type III humanized collagen and the recombinant type XVII humanized collagen are removed and replaced with an equal weight of glycerol, and the other components remain unchanged;

[0075] Group B: Compared with Example 2, recombinant humanized type I collagen and recombinant humanized type XVII collagen were removed and replaced with an equal weight of glycerol, and the other components remained unchanged;

[0076] Group C: The difference from Example 2 is that the recombinant humanized type I collagen and recombinant humanized type III collagen were removed and replaced with glycerol of equal weight, and the other components remained unchanged;

[0077] Group D: The difference from Example 2 is that the recombinant humanized type XVII collagen was removed and replaced with an equal weight of glycerol, and the other components remained unchanged;

[0078] Group E: The difference from Example 2 is that the recombinant humanized type III collagen was removed and replaced with an equal weight of glycerol, and the other components remained unchanged;

[0079] Group F: The difference from Example 2 is that the recombinant humanized type I collagen was removed and replaced with an equal weight of glycerol, and the other components remained unchanged;

[0080] Group G: The difference from Example 2 is that the sea fennel water extract, Antrodia camphorata extract, Hippophae rhamnoides extract, Bistorta root extract, and Dendrobium candidum extract are removed and replaced with glycerin of equal weight, and the other components remain unchanged;

[0081] Group H: The difference from Example 2 is that the content of each component is different. Specifically, every 100 mL of the composition includes: 2.5 mg recombinant humanized type I collagen, 12.5 mg recombinant humanized type III collagen, 2.5 mg recombinant type XVII humanized collagen, 1.00 mg ectoine, 0.50 mg ergothioneine, 0.63 mg niacinamide and 0.45 mg sea fennel water extract, 0.40 mg Centella asiatica extract, 0.30 mg Antrodia cinnamomea extract, 0.25 mg Hippophae rhamnoides extract, 0.30 mg Polygonum multiflorum root extract, 0.20 mg Dendrobium officinale extract, 0.25 mg retinol, and 0.15 mg erythritol.

[0082] Test Example 1

[0083] Evaluation of cell proliferation effect: HaCaT cells (human immortalized keratinocytes) in good growth state were plated on a 96-well plate and cultured for 17 hours. After replacing the serum-free basal medium and culturing for 7 hours, the old medium was discarded, and a medium containing the composition (2 μg / mL) prepared in Examples 1-4 or Comparative Example AH group was added. The culture medium of the blank group did not contain the composition, and the rest was consistent with the experimental group. After culturing for 48 hours, the cell viability was detected by the CCK8 method; the absorbance value was measured at a wavelength of 450 nm, each data was tested three times, and the average value of each group was calculated. The measurement results are shown in Table 1.

[0084] The calculation formula is:

[0085] Relative cell proliferation rate (%) = (absorbance of experimental group - absorbance of blank group) / absorbance of blank group × 100%.

[0086] Table 1 Relative cell proliferation rate test results

[0087] Group Relative cell proliferation rate (%) Blank Group 0.01 Example 1 78.5 Example 2 83.7 Example 3 80.4 Example 4 76.9 Comparative Example A 6.7 Comparative Example B 8.3 Comparative Example C 1.7 Comparative Example D 10.9 Comparative Example E 15.4 Comparative Example F 18.5 Comparative Example G 20.3 Comparative Example H 27.1

[0088] It can be seen from the data in Table 1 that the wrinkle-lightening and firming triple active collagen compositions of Examples 1-4 have excellent effects on promoting cell proliferation; compared with Example 2, the raw materials of Comparative Examples AF lack recombinant type I, type III, and type XVII humanized collagen, respectively, and their relative cell proliferation rates are significantly lower than those of Example 2, indicating that recombinant type I, type III, and type XVII humanized collagen play a major role in promoting cell proliferation; and compared with Example 2, Comparative Example G also lacks sea fennel water extract, camphor extract, sea buckthorn extract, Polygonum multiflorum root extract, Dendrobium officinale extract and other substances, so the relative cell proliferation rate is lower, indicating that plant extracts can also promote cell proliferation; Compared with Example 2, the contents of various components in Comparative Example H are different, all of which are increased but not within the appropriate component range given by the present invention, so the relative cell proliferation rate is also lower, but higher than Comparative Example AG, because all components are complete and still have a certain effect of promoting cell proliferation. It can be seen that the components in the wrinkle-lightening and firming triple active collagen composition of the present invention can work together to better promote cell proliferation only after being reasonably matched, and the components synergistically enhance the cell proliferation ability.

[0089] Test Example 2

[0090] Evaluation of fibroblast migration effect: Normally grown NIH-3T3 mouse fibroblasts were digested and inoculated into 6-well plates at 2×10 cells per well. 6The cells were cultured at 37°C until the cells were confluent in a 96-well plate. The complete DMEM high-glucose medium in the 96-well plate inoculated with NIH-3T3 cells was aspirated, scratched with a 10 μL pipette tip, and the cell fragments were washed 3 times with PBS. The control group used a complete medium DMEM, and the experimental group used a complete medium containing the composition (2 μg / mL) prepared in Examples 1-4 or Comparative Example AH group. After culturing for 24 hours, the cell migration distance was measured and the cell migration rate was calculated. The measurement results are shown in Table 2;

[0091] Cell migration rate (%) = (scratch width at 0 h - scratch width at 24 h) / scratch width at 0 h × 100%.

[0092] Table 2 Results of fibroblast migration rate assay

[0093] Group Cell migration rate (%) Blank Group 60.7 Example 1 93.4 Example 2 134.8 Example 3 113.7 Example 4 108.9 Comparative Example A 24.7 Comparative Example B 25.4 Comparative Example C 27.8 Comparative Example D 20.1 Comparative Example E 31.7 Comparative Example F 28.5 Comparative Example G 47.3 Comparative Example H 55.8

[0094] As can be seen from Table 2, the wrinkle-lightening and firming triple active collagen compositions of Examples 1-4 have the effect of promoting cell migration; compared with Example 2, the raw materials of Comparative Examples AF lack recombinant type I, type III, and type XVII humanized collagen, respectively, and their relative cell proliferation rates are significantly lower than those of Example 2, indicating that recombinant type I, type III, and type XVII humanized collagen play a major role in promoting cell migration; and compared with Example 2, Comparative Example G also lacks sea fennel water extract, Antrodia camphorata extract, Hippophae rhamnoides extract, Polygonum multiflorum root extract, Dendrobium officinale extract and other substances, so the relative cell migration rate is lower, indicating that plant extracts can also promote cell migration; Compared with Example 2, the content of each component in Comparative Example H is different, all of which are improved but not within the appropriate component range given by the present invention, so the relative cell migration rate is also lower, but higher than Comparative Example AG, because all components are complete and still have a certain effect of promoting cell migration. Since the strength of cell migration ability can reflect the strength of cell activity, and the compositions of the present invention all have the effect of promoting cell migration, it can be shown that the compositions of the present invention can achieve anti-aging effects by promoting cell activity and polarity.

[0095] Test Example 3

[0096] 90 volunteers (women aged 23 to 40 years old) were selected to participate in the anti-aging effect test of the composition of the present invention. The volunteers used the compositions prepared by Example 2 and Comparative Examples AH once in the morning and evening every day for 1 month. Before using the essence and after using it on the 0th, 14th and 28th day, the skin firmness, skin elasticity and the number and area of ​​crow's feet on one random cheek were measured. The measurement results are shown in Tables 3-5.

[0097] Table 3 Determination results of skin firmness F4 value

[0098] Group Before use Use for 14 days Use for 28 days Improvement rate after 28 days % Example 2 3.90 3.43 2.78 28.72 Comparative Example A 3.96 3.81 3.75 5.30 Comparative Example B 4.07 4.00 3.93 3.44 Comparative Example C 4.16 4.13 4.00 3.85 Comparative Example D 4.09 4.05 3.94 3.67 Comparative Example E 3.93 3.84 3.78 3.82 Comparative Example F 3.99 3.91 3.85 3.51 Comparative Example G 3.97 3.73 3.64 8.31 Comparative Example H 3.90 3.62 3.32 14.87

[0099] Note: Improvement rate (%) = (test value before use - test value after using the sample for N days) / test value before use × 100%.

[0100] The smaller the F4 value, the firmer the skin. As shown in Table 3, compared with before use, after using the composition of Example 2 for 14 days and 28 days, the skin firmness F4 parameter was significantly improved, and decreased by 12.05% and 28.72% respectively; while the skin firmness of the comparative example AH group was also improved to a certain extent, but the improvement was significantly lower than the effect of the composition of Example 2 of the present invention. This shows that the use of the composition of the present invention can make the skin firmer.

[0101] Table 4 Measurement results of skin elasticity R2 value

[0102] Group Before use Use for 14 days Use for 28 days Improvement rate after 28 days % Example 2 0.59 0.62 0.69 16.95 Comparative Example A 0.57 0.58 0.59 3.51 Comparative Example B 0.59 0.60 0.61 3.39 Comparative Example C 0.58 0.58 0.59 1.72 Comparative Example D 0.55 0.56 0.57 3.64 Comparative Example E 0.58 0.60 0.61 5.17 Comparative Example F 0.61 0.63 0.64 4.92 Comparative Example G 0.56 0.58 0.59 5.36 Comparative Example H 0.54 0.57 0.59 9.26

[0103] Note: Improvement rate (%) = (test value after using the sample for N days - test value before use) / test value before use × 100%.

[0104] The larger the R2 value, the better the skin elasticity. As shown in Table 3, compared with before use, after using the composition of Example 2 for 14 days and 28 days, the skin elasticity R2 parameter was significantly improved by 5.08% and 16.95% respectively; while the skin elasticity of the comparative example AH group was also improved to a certain extent, but the improvement was significantly lower than the effect of the composition of Example 2 of the present invention. This shows that the use of the composition of the present invention can make the skin more elastic.

[0105] Table 5 Measurement results of the number of crow's feet

[0106] Group Before use Use for 14 days Use for 28 days Improvement rate after 28 days % Example 2 87 81 70 19.54 Comparative Example A 78 77 75 3.85 Comparative Example B 84 83 81 3.57 Comparative Example C 85 84 83 2.35 Comparative Example D 79 77 76 3.80 Comparative Example E 74 72 70 5.41 Comparative Example F 71 69 67 5.63 Comparative Example G 77 75 72 6.49 Comparative Example H 78 74 71 8.97

[0107] Note: Improvement rate (%) = (test value before use - test value after using the sample for N days) / test value before use × 100%.

[0108] The smaller the measured value, the fewer the number of crow's feet. It can be seen from Table 3 that compared with before use, the number of crow's feet was significantly improved after using the composition of Example 2 for 14 days and 28 days, which was significantly reduced by 6.90% and 19.54%, respectively; although the comparative example AH group also improved the number of crow's feet to a certain extent, the degree of improvement was significantly lower than the effect of the composition of Example 2 of the present invention.

[0109] Table 6 Measurement results of crow's feet area

[0110] Group Before use Use for 14 days Use for 28 days Improvement rate after 28 days % Example 2 48.93 45.25 38.80 20.70 Comparative Example A 41.35 40.15 39.83 3.68 Comparative Example B 43.06 42.11 41.35 3.97 Comparative Example C 45.85 45.25 44.18 3.64 Comparative Example D 47.25 46.26 44.63 5.54 Comparative Example E 42.87 41.93 40.48 6.27 Comparative Example F 47.06 45.67 44.54 5.35 Comparative Example G 49.43 47.20 45.95 7.04 Comparative Example H 46.94 44.34 41.89 10.76

[0111] Note: Improvement rate (%) = (test value before use - test value after using the sample for N days) / test value before use × 100%.

[0112] The smaller the measured value, the smaller the crow's feet area. As shown in Table 3, compared with before use, the area of ​​crow's feet has been significantly improved after using the composition of Example 2 for 14 days and 28 days, which is significantly reduced by 7.52% and 20.70% respectively; while the comparative example AH group also improved the area of ​​crow's feet to a certain extent, but the improvement degree is significantly lower than the effect of the composition of Example 2 of the present invention. It can be shown that the composition of the present invention has the effect of reducing facial fine lines.

[0113] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0114] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A triple active collagen composition for lightening and firming wrinkles, characterized in that: Each 100 mL of the composition includes: 0.5-2 mg recombinant humanized collagen type I; 2.5-10 mg recombinant humanized collagen type III; 0.5-2 mg recombinant humanized collagen type XVII; 0.25-0.8 mg ectoine; 0.12-0.46 mg ergothioneine; 0.32-0.51 mg niacinamide; 0.15-0.40 mg sea fennel water extract; 0.10-0.37 mg Centella asiatica extract; 0.12-0.28 mg Antrodia cinnamomea extract; 0.08-0.17 mg Hippophae rhamnoides extract; 0.05-0.15 mg Polygonum multiflorum root extract; 0.01-0.10 mg Dendrobium officinale extract; 0.01-0.15 mg retinol; and 0.01-0.08 mg erythritol; The triple active collagen composition for reducing wrinkles and tightening skin is used for reducing wrinkles and tightening skin.

2. The wrinkle-lightening and firming triple active collagen composition according to claim 1, characterized in that: The preparation method of the sea fennel water extract comprises the following steps: crushing the sea fennel, passing through a 40-mesh sieve, adding the sea fennel powder into distilled water according to a ratio of sea fennel to distilled water (m:v)=1:8, stirring evenly, performing water bath ultrasonic extraction, filtering, centrifuging, concentrating the supernatant, and drying to obtain the sea fennel extract.

3. The wrinkle-lightening and firming triple active collagen composition according to claim 2, characterized in that: In the preparation method of sea fennel water extract, the water bath temperature is 50° C., the ultrasonic power is 500 W, and the extraction is performed for 2 hours.

4. The wrinkle-lightening and firming triple active collagen composition according to claim 2, characterized in that: In the preparation method of sea fennel water extract, the centrifugation is performed at 800 rpm for 5 minutes.

5. The wrinkle-lightening and firming triple active collagen composition according to claim 1, characterized in that: The camphorata extract is a water extract.

6. The wrinkle-lightening and firming triple active collagen composition according to claim 5, characterized in that: The preparation method of the water extract of Antrodia cinnamomea is as follows: crush Antrodia cinnamomea, pass through a 0.42 mm sieve, add Antrodia cinnamomea powder into distilled water in a ratio of Antrodia cinnamomea: distilled water (m:v=1:10), stir evenly, extract by heating in a water bath, filter, centrifuge, concentrate and dry the supernatant to obtain the Antrodia cinnamomea extract.

7. The wrinkle-lightening and firming triple active collagen composition according to claim 6, characterized in that: In the preparation method of the water extract of Antrodia cinnamomea, the water bath temperature is 35° C. and the extraction time is 8 hours.

8. The wrinkle-lightening and firming triple active collagen composition according to claim 6, characterized in that: In the preparation method of the aqueous extract of Antrodia cinnamomea, the centrifugation is performed at 500 rpm for 10 minutes.

9. The method for preparing the wrinkle-lightening and firming triple active collagen composition according to any one of claims 1 to 8, characterized in that: The following steps are involved: (1) mixing recombinant humanized type I collagen, recombinant humanized type III collagen and recombinant humanized type XVII collagen to obtain multiple humanized collagens; (2) mixing ectoine, ergothioneine, sea fennel extract, Centella asiatica extract and water, heating and stirring to dissolve evenly, and keeping the temperature to obtain a mixed solution A; (3) After the mixed solution A is cooled, niacinamide, retinol and erythritol are added in sequence and heated until completely dissolved to obtain a mixed solution B; (4) After the mixed solution B cools down, add multiple humanized collagen, Antrodia camphorata extract, Hippophae rhamnoides extract, Polygonum multiflorum root extract and Dendrobium officinale extract in sequence, stir evenly and homogenize to obtain a wrinkle-lightening and firming triple active collagen composition.

10. The preparation method according to claim 9, characterized in that: In step (2), the heating temperature is 65-75°C, the insulation time is 15-30 min, the stirring speed is 300-400 rpm, and the stirring time is 5-10 min; in step (3), the cooling is 50-60°C, the heating time is 15-20 min, the stirring speed is 550-650 rpm, and the stirring time is 3-8 min.

11. The preparation method according to claim 9, characterized in that: In step (4), the cooling speed is 35-40°C, the stirring speed is 400-500 rpm, and the stirring time is 5-15 min; the homogenization speed is 2100-2800 rpm, and the homogenization time is 8-12 min.

12. Use of the wrinkle-lightening and firming triple active collagen composition according to any one of claims 1 to 8, characterized in that: The use is in the preparation of wrinkle-lightening and firming cosmetics or medicines.

13. Use of the wrinkle-lightening and firming triple active collagen composition prepared by the preparation method according to any one of claims 9 to 11, characterized in that: The use is in the preparation of wrinkle-lightening and firming cosmetics or medicines.

14. Use according to claim 12 or 13, characterized in that The wrinkle-lightening and firming triple-active collagen composition improves cell vitality and promotes cell proliferation.

15. Use according to claim 12 or 13, characterized in that The wrinkle-lightening and firming triple-active collagen composition promotes skin firmness and elasticity.