CN109700687B

CN109700687B - Flexible liposome cosmetic containing active polypeptide and preparation method thereof

Info

Publication number: CN109700687B
Application number: CN201910094274.4A
Authority: CN
Inventors: 杨莉; 魏于全
Original assignee: Shenzhen Gaoshang Kemei Biotechnology Co ltd
Current assignee: Shenzhen Gaoshang Kemei Biotechnology Co ltd
Priority date: 2018-02-13
Filing date: 2019-01-30
Publication date: 2022-04-15
Anticipated expiration: 2039-01-30
Also published as: CN109700687A

Abstract

The invention belongs to the field of cosmetics, and particularly relates to a flexible liposome cosmetic containing active polypeptide and a preparation method thereof. The invention provides a flexible liposome cosmetic containing active polypeptide aiming at the problems that most active ingredients in the cosmetic containing the active polypeptide are difficult to permeate the skin due to the barrier effect of the horny layer of the epidermis of the skin, the flexible liposome modified by hydrophobic modified polypeptide contains the active polypeptide to prepare the cosmetic, the permeability of the active polypeptide in the cosmetic is effectively improved, the cosmetic with better absorption, oxidation resistance and anti-aging effects is developed, and the application prospect is wide.

Description

Flexible liposome cosmetic containing active polypeptide and preparation method thereof

Technical Field

The invention belongs to the field of cosmetics, and particularly relates to a flexible liposome cosmetic containing active polypeptide and a preparation method thereof.

Background

Polypeptides are compounds in which two or more amino acids are condensed and joined by peptide bonds, which are intermediates of proteins. A peptide containing less than 10 amino acids is called an oligopeptide, more than 50 amino acids is called a polypeptide, and a polypeptide is called a protein. The polypeptide substance has high biological activity, and can regulate various physiological activities and biochemical reactions of body cells, and is also called active biological polypeptide.

In the defense and care process of skin natural aging, the active biological polypeptide also plays unique and important physiological roles, such as proliferation of skin tissue cells, cell chemotaxis and migration, repair and regeneration, angiogenesis and reconstruction, pigment formation and removal, protein synthesis and secretion, metabolism and regulation and the like.

Currently, most of the active polypeptides related to skin beauty, which are widely used, are chemically synthesized peptides (peptides, CPs) with small molecular weight, which can be classified into three categories according to their functions: 1) anti-wrinkle, which can block the signal of facial nerve transmission muscle contraction and affect the skin sac nerve conduction, and relax facial muscles, and improve dynamic and static wrinkles and fine lines of facial skin, and comprises acetyl hexapeptide-8 (also known as Argireline, ayurrelin), palmitoyl tripeptide, acetyl octapeptide, etc.; 2) anti-free radical species, which contribute to anti-inflammatory and antioxidant properties of the skin, including palmitoyl tetrapeptide-7, glutathione, carnosine, L-carnosine, and the like; 3) promoting cell activity, promoting synthesis of collagen and glucosamine in human fibroblast, strengthening connective tissue, increasing skin compactness, and promoting cell activity, including palmitoyl pentapeptide-3, palmitoyl pentapeptide-4, copper peptide, palmitoyl tripeptide-1, palmitoyl tripeptide-5, acetyl tetrapeptide-9, and myristoyl hexapeptide-4; 4) whitening skin, relieving swelling, promoting eyelash growth, inhibiting melanocyte melanin production, or as hygroscopic agent to relieve eye edema, or activating keratin gene expression to promote eyelash growth, including nonapeptide-1, dipeptide-2, acetyl tetrapeptide-5, and myristoyl pentapeptide-17. And the like.

The molecular weight of these active polypeptides is much reduced compared to proteins, and some are hydrophobically modified, but most of the active ingredients are difficult to pass through due to the barrier effect of the stratum corneum of the skin epidermis. Therefore, it is of great significance to develop cosmetics capable of transdermal delivery of active substances having antioxidant and anti-aging effects, such as active polypeptides, with high efficiency.

Disclosure of Invention

The object of the present invention is to provide a cosmetic comprising flexible liposomes of active polypeptides.

The cosmetic containing the flexible liposome of the active polypeptide is prepared by hydrophobic modification of the flexible liposome containing the active polypeptide.

Furthermore, in the cosmetic comprising the flexible liposome of active polypeptide, the hydrophobic modified polypeptide is a nitrogen-terminal hydrophobic modified polypeptide.

In the cosmetic containing the flexible liposome of the active polypeptide, the flexible liposome modified by the hydrophobic modified polypeptide is prepared from the following raw materials in parts by weight: lecithin-sodium deoxycholate or polysorbate 80 derivative 7:2-4, 1-10 wt% of hydrophobic modified polypeptide, and active polypeptide.

Wherein, the cosmetic of the flexible liposome containing the active polypeptide also contains an antioxidant. The antioxidant comprises: at least one of vitamin C or its derivative, vitamin E or its derivative, or coenzyme Q. The weight percentage of the antioxidant is 0.1-1%.

In the cosmetic containing the flexible liposome of the active polypeptide, the active polypeptide is an active polypeptide which can be used in the field of cosmetics. The active polypeptide used in the field of cosmetics is an active polypeptide having at least one function of anti-wrinkle, anti-free radical, cell activity promotion, whitening, detumescence or eyelash growth promotion.

Specifically, the peptide comprises at least one of copper peptide, dipeptide-2, palmitoyl tripeptide-1, palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, acetyl tetrapeptide-5, acetyl tetrapeptide-9, palmitoyl pentapeptide-3, palmitoyl pentapeptide-4, acetyl hexapeptide-8, acetyl octapeptide, myristoyl pentapeptide-17, myristoyl hexapeptide-4, nonapeptide-1, carnosine, L-carnosine and glutathione.

Further, the weight percentage of the active polypeptide is 0.01-1%.

Specifically, the composition of the polypeptide is as follows:

acetyl hexapeptide-8: Ac-Glu-Glu-Met-Gln-Arg-Arg-NH₂；

Acetyl tetrapeptide-5: Ac-beta-Ala-His-Ser-His-OH;

palmitoyl pentapeptide-4: PAL (palmitic acid) -Lys-Thr-Thr-Lys-Ser-OH;

palmitoyl tetrapeptide-7: PAL (palmitic acid) -Gly-Gln-Pro-Arg-OH;

palmitoyl tripeptide-1: PAL (palmitic acid) -Gly-His-Lys-OH;

myristoyl pentapeptide-17: Myr-Lys-Leu-Ala-Lys-Lys-Lys-NH₂；

Carnosine: beta-Ala-L-His;

glutathione: NH 2-Glu-Cys-Gly-OH;

nonapeptide-1: Met-Pro-D-Phe-Arg-D-Trp-Phe-Lys-Pro-Val-NH₂。

Amino acid sequence of SEQ ID NO 2 nonapeptide-1

MPFRWFKPV。

Furthermore, the cosmetic containing the flexible liposome of the active polypeptide is prepared from the following components in proportion: lecithin sodium deoxycholate or polysorbate 80 derivative 7:3, 0.5 wt% antioxidant, and 2-10 wt% hydrophobic modified polypeptide.

Furthermore, in the cosmetics containing the flexible liposome of the active polypeptide, the sequence of the polypeptide in the hydrophobic modified polypeptide is that NH is connected to the C end on the basis of SEQ ID NO. 1₂And coupling sterol compounds or saturated straight-chain fatty acids at the nitrogen terminal of the polypeptide.

1 amino acid sequence of hydrophobic modified polypeptide of SEQ ID NO

VQWRIRVAVIRK。

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the sterol compound is a cholesterol compound or a cholic acid compound.

Furthermore, in the cosmetic containing the flexible liposome of active polypeptide, the sterol compound is at least one of cholesterol, succinylated cholesterol, cholic acid or deoxycholic acid.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the saturated straight-chain fatty acid is at least one of C6-C20.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the saturated straight-chain fatty acid is at least one of C8-C18.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the long-chain fatty acid is at least one of stearic acid, palmitic acid, lauric acid and n-caprylic acid.

Furthermore, in the cosmetic containing the flexible liposome of active polypeptide, the hydrophobic modified polypeptide has the following structure:

wherein, R is a sterol compound or saturated straight chain fatty acid.

Furthermore, in the cosmetics containing the flexible liposome of the active polypeptide, R is

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the anti-wrinkle active polypeptide is at least one of palmitoyl tripeptide, acetyl hexapeptide-8 or acetyl octapeptide.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the anti-wrinkle polypeptide accounts for 0.01-3% of the weight of the liposome.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the anti-wrinkle polypeptide accounts for 0.05-2% of the weight of the liposome.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the anti-wrinkle polypeptide accounts for 0.1-1% of the liposome by weight.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the anti-free radical polypeptide is at least one of glutathione, palmitoyl tetrapeptide-7, carnosine or L-carnosine.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the anti-free radical polypeptide accounts for 0.01-3% of the weight of the liposome.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the anti-free radical polypeptide accounts for 0.05-2% of the weight of the liposome.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the anti-free radical polypeptide accounts for 0.1-1% of the weight of the liposome.

Furthermore, in the cosmetic containing the flexible liposome of active polypeptide, the active polypeptide for promoting cell activity is at least one of palmitoyl pentapeptide-3, palmitoyl pentapeptide-4, copper peptide, palmitoyl tripeptide-1, palmitoyl tripeptide-5, acetyl tetrapeptide-9 or myristoyl hexapeptide-4.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the weight percentage of the active polypeptide for promoting cell activity accounts for 0.01-3% of the liposome.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the weight percentage of the active polypeptide for promoting cell activity accounts for 0.05-2% of the liposome.

Furthermore, in the cosmetic containing the flexible liposome of active polypeptide, the weight percentage of the active polypeptide for promoting cell activity in the liposome is 0.1-1%.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the whitening active polypeptide is nonapeptide-1.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the detumescence active polypeptide is acetyl tetrapeptide-5 or dipeptide-2.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the active polypeptide for promoting eyelash growth is myristoyl pentapeptide-17.

Furthermore, the flexible liposome cosmetic containing the active polypeptide has the active polypeptide with the whitening, detumescence and eyelash growth promoting effects accounting for 0.01-3 wt% of the liposome.

Furthermore, the weight percentage of the whitening, detumescence and eyelash growth promoting active polypeptide in the flexible liposome cosmetic containing the active polypeptide is 0.05-2%.

Furthermore, the flexible liposome cosmetic containing the active polypeptide has the active polypeptide with the whitening, detumescence and eyelash growth promoting effects accounting for 0.1-1 wt% of the liposome.

Further, in the above cosmetic composition comprising flexible liposomes containing an active polypeptide, the cosmetic composition further comprises at least one of an emulsifier, a co-emulsifier, a skin conditioner, a whitening agent, a colorant, a humectant, a solubilizer, a surfactant, a preservative, a fragrance, an emollient, an anti-acne agent, a film-forming agent, a thickener, a pH adjuster, a buffer, a stabilizer, or an ultraviolet absorber, which are used in the preparation of the cosmetic composition.

Furthermore, in the cosmetic containing the flexible liposome of the active polypeptide, the dosage form of the cosmetic containing the flexible liposome of the active polypeptide is water, emulsion, paste, jelly, troche or aerosol.

Furthermore, in the cosmetic containing the flexible liposome of active polypeptide, the dosage form of the cosmetic containing the flexible liposome of active polypeptide is emulsion, essence, cream, mask, freeze-dried powder or facial cleanser.

The invention also provides a method for preparing the flexible liposome cosmetic containing the active polypeptide, which comprises the following steps:

a. weighing lecithin, sodium deoxycholate or polysorbate 80 derivatives and vitamin E, placing into a reaction container, adding solvent for dissolving, wherein the solvent is a mixture of chloroform and ethanol at a ratio of 1: 1-3 or is ethanol;

b. vacuum rotary evaporating to obtain liposome membrane, and drying;

c. adding hydrophobic modified polypeptide and distilled water into the liposome membrane, and performing ultrasonic hydration to obtain a blank liposome solution;

d. adding the active polypeptide into the blank liposome solution, slightly shaking, and incubating at room temperature for 30-60 min to obtain the flexible liposome containing the active polypeptide.

Wherein, the method for preparing the flexible liposome cosmetic containing the active polypeptide further comprises the following steps of e: and extruding the liposome subjected to vacuum decompression through a 0.1-0.45 mu m polycarbonate membrane for 4-8 times to obtain a blank liposome solution.

Wherein, the method for preparing the flexible liposome cosmetic containing the active polypeptide further comprises the following steps of f: according to the dosage form requirement, the flexible liposome is further added with at least one of emulsifying agent, auxiliary emulsifying agent, skin conditioning agent, whitening agent, colorant, humectant, solubilizer, surfactant, antiseptic, aromatic, emollient, anti-acne agent, film forming agent, thickener, pH regulator, buffer, stabilizer or ultraviolet absorbent to prepare cosmetic dosage form.

The cosmetic is a daily chemical industrial product which is applied to any part (skin, hair, nails, lips and the like) of the surface of a human body by smearing, spraying or other similar methods so as to achieve the purposes of cleaning, eliminating bad smell, protecting skin, beautifying and decorating.

The flexible liposome is a self-aggregating vesicle improved by prescription on the basis of liposome, and a surface active substance such as sodium cholate, polysorbate 80 or derivatives thereof and the like is added into the phospholipid component of the liposome, so that the lipid membrane of the liposome has high deformability, and the liposome is also called a transfersome. The carrier has smaller particle size than liposome, and can pass through small holes with the pore size of 1/10-1/5, and the permeation rate and the permeation quantity are almost equal to those of pure water.

The action mechanism of the flexible liposome (transfersome) of the invention is as follows: 1. the particle size of the carrier is smaller than that of the liposome, and the carrier can quickly penetrate through the horny layer of the skin and enter the epidermis and dermis to form a reservoir; 2. the skin active ingredients and water encapsulated in the transfer body can be slowly released, and the action effect of the active substances is greatly improved. 3. The carrier carries encapsulated skin active polypeptide, such as palmitoyl tripeptide or acetyl hexapeptide-3, which can block signals of facial nerve transmission muscle contraction, and has anti-wrinkle effect; or palmitoyl pentapeptide-3, palmitoyl tripeptide-1 or acetyl tetrapeptide-9, etc. can promote the synthesis of collagen and glucosamine in human fiber cells, strengthen connective tissues and play a role in repairing skin; or palmitoyl tetrapeptide-7, carnosine, etc. with effects of scavenging free radicals, relieving inflammation and resisting oxidation; or nonapeptide-1, etc. can inhibit melanocyte from producing melanin, and has skin whitening effect; or acetyl tetrapeptide-5, dipeptide-2, etc. can eliminate edema and has the function of eliminating eye bags; or myristoyl pentapeptide-17, etc. can activate human keratin gene expression and promote eyelash growth.

Furthermore, the active polypeptide-containing carrier of the present invention is compounded with an emulsifier, a co-emulsifier, a skin conditioner, a whitening agent, a colorant, a humectant, a solubilizer, a surfactant, a preservative, an aromatic, an emollient, an anti-acne agent, a film-forming agent, a thickener, a pH adjuster, a buffer, a stabilizer, an ultraviolet absorber, etc. in accordance with the art of the art, the active polypeptide-containing carrier can be prepared into cosmetics in the form of a water aqua, an emulsion, an essence, a cream, a mask, a lyophilized powder, a facial cleanser, etc. Various products of the present invention may be applied to the human face (including eyes), neck, hands and feet, and the skin of the whole body.

The invention has the beneficial effects that:

the invention provides a novel flexible liposome based on the traditional liposome through prescription improvement, surface active substances such as sodium deoxycholate, polysorbate 80 or derivatives thereof and the like are added into phospholipid components of the liposome, so that a lipid membrane of the liposome has high deformability, and meanwhile, cell-penetrating peptide capable of enhancing transdermal effect is added, so that the transdermal permeability of the liposome is improved. The inventor uses the transfersome to wrap active polypeptide, uses its excellent transdermal ability to prepare new active polypeptide transdermal agent, and develops its application in cosmetic field. The novel active polypeptide transdermal agent is safe and nontoxic in material, and has good effects in the aspects of wrinkle resistance, aging resistance, oxidation resistance, repair and the like. In addition, it has the functions and characteristics of biological membrane, strong affinity with human body cell, unique efficacy in cosmetics and great economic benefit.

Preliminary tests show that: the carrier carries encapsulated active polypeptide, such as palmitoyl tripeptide or acetyl hexapeptide-3, which can block signals of facial nerve transmission muscle contraction, and has anti-wrinkle effect; the encapsulation of palmitoyl pentapeptide-3, palmitoyl tripeptide-1 or acetyl tetrapeptide-9 and the like can promote the synthesis of collagen and glucosamine in human body fiber cells, strengthen connective tissues and play a role in repairing skin; the encapsulated palmitoyl tetrapeptide-7, carnosine and the like can resist free radicals, and have the effects of resisting inflammation and resisting oxidation; or nonapeptide-1, etc. can inhibit melanocyte from producing melanin, and has skin whitening effect; or acetyl tetrapeptide-5, dipeptide-2, etc. can eliminate edema and has the function of eliminating eye bags; or myristoyl pentapeptide-17, etc. can activate human keratin gene expression and promote eyelash growth.

On the technical level, the invention has the advantages that: besides the general functions of resisting aging, moisturizing and promoting softness in the market, the product of the invention also applies a novel transdermal technology to cosmetics, and has better effect.

Drawings

FIG. 1 is a graph of prepared nanoparticle size measurements of DP7-C modified flexible liposomes containing acetyl hexapeptide-8 with polysorbate 80 as a surfactant;

FIG. 2 is a nano-particle size assay of prepared acetyl hexapeptide-8-containing PAL-DP7 modified flexible liposomes with polysorbate 80 as a surfactant;

FIG. 3 is a graph showing the prepared DP7-C modified flexible liposome nanoparticle size assay with palmitoyl tetrapeptide-7 using polysorbate 80 as a surfactant;

FIG. 4 is a graph showing the particle size of PAL-DP7 modified flexible liposomes containing palmitoyl tetrapeptide-7 prepared using polysorbate 80 as a surfactant;

FIG. 5 is a graph showing the prepared DP7-C modified flexible liposome nanoparticle size assay with palmitoyl pentapeptide-4 using polysorbate 80 as a surfactant;

FIG. 6 is a nano-particle size test chart of PAL-DP7 modified flexible liposomes containing palmitoyl pentapeptide-4 prepared using polysorbate 80 as a surfactant;

FIG. 7 is a graph showing the particle size of PAL-DP7 modified flexible liposomes containing nonapeptide-1 prepared using polysorbate 80 as a surfactant;

FIG. 8 is a graph showing the prepared DP7-C modified flexible liposome nanoparticle size assay with nonapeptide-1 using polysorbate 80 as a surfactant;

FIG. 9 is a graph of DP7-C modified flexible liposome nanoparticle size assay prepared with polysorbate 80 as the surfactant and containing acetyl tetrapeptide-5;

FIG. 10 is a graph showing the nanoparticle size detection of PAL-DP7 modified flexible liposomes containing acetyl tetrapeptide-5 prepared with polysorbate 80 as a surfactant;

fig. 11 is a graph of prepared nanoparticle size detection of myristic pentapeptide-17-containing DP7-C modified flexible liposomes with polysorbate 80 as the surfactant;

FIG. 12 is a graph showing the particle size of prepared PAL-DP7 modified flexible liposomes containing myristoyl pentapeptide-17 with polysorbate 80 as a surfactant;

FIG. 13 is a graph showing the in vitro transdermal effect of DP7-C and PAL-DP7 modified flexible liposomes containing acetyl hexapeptide-8 prepared;

FIG. 14 is a graph showing the in vitro transdermal effect of the prepared palmitoyl tetrapeptide-7-containing DP7-C and PAL-DP7 modified flexible liposomes;

FIG. 15 is a graph showing the in vitro transdermal effect of the prepared palmitoyl pentapeptide-4 containing DP7-C and PAL-DP7 modified flexible liposomes;

FIG. 16 is a graph showing the in vitro transdermal effect of prepared nonapeptide-1 containing DP7-C and PAL-DP7 modified flexible liposomes;

FIG. 17 is a graph showing the in vitro transdermal effect of DP7-C and PAL-DP7 modified flexible liposomes containing acetyl tetrapeptide-5 prepared;

FIG. 18 is a graph showing the in vitro transdermal effect of DP7-C and PAL-DP7 modified liposomes containing myristoyl pentapeptide-17 prepared.

Detailed Description

The hydrophobic modified polypeptide DP7-C is a conjugate of antimicrobial peptide DP7 and cholesterol, the hydrophobic modified polypeptide PAL-DP7 is a conjugate of antimicrobial peptide DP7 and palmitic acid, the synthetic method is disclosed in patent CN107441501A, and other methods in the field can be adopted for synthesis, and DP7-C and PAL-DP7 used in the examples are synthesized by Dorkaee biological medicine science and technology development Co.

The raw materials and equipment used in the embodiment of the present invention are known products and obtained by purchasing commercially available products.

EXAMPLE 1 preparation of Flexible liposomes containing active Polypeptides of the invention

Precisely weighing soybean lecithin, sodium deoxycholate or polysorbate 80 and vitamin E according to the formula, dissolving with chloroform/ethanol or ethanol respectively, dissolving and mixing in a 250ml pear-shaped bottle, rotary evaporating at room temperature for 2 hours, and placing the obtained film in a vacuum drying oven overnight. Distilled water and a proper amount of hydrophobic modified polypeptide are added in the next day, and a 400w probe is used for ultrasonic treatment for 30min to obtain a blank carrier solution. Slowly adding a proper amount of acetyl octapeptide-8, palmitoyl tetrapeptide, palmitoyl pentapeptide, nonapeptide-1, acetyl tetrapeptide-5, myristoyl pentapeptide-17 and the like, incubating at room temperature for 40 minutes, filtering the obtained mother liquor for multiple times by using a 0.2-micron polycarbonate membrane, adding an excipient of 5% mannitol, subpackaging in penicillin bottles, and freeze-drying to obtain the flexible liposome containing the active polypeptide.

Experimental example 1-1 preparation of DP7-C modified Flexible liposomes containing acetyl hexapeptide-8

Experimental materials:

soybean lecithin: purchased from Lipoid GmbH;

sodium deoxycholate: purchased from biotechnology responsibility ltd of obozoxing, beijing;

polysorbate 80: from Merck, USA;

hydrophobic modified polypeptide DP 7-C: the Chengdayaki biological medicine science and technology development company is synthesized;

acetyl hexapeptide-8: the Chengdayaki biological medicine science and technology development company is synthesized;

the instrument comprises the following steps: malvern laser particle size detector ZEN 3600.

In the experimental example, the flexible liposome is modified by DP7-C, the surface active substances are respectively prepared by sodium deoxycholate and polysorbate 80, and the preparation process of the flexible liposome containing acetyl hexapeptide-8 has the following parameters: the proportion of the soybean lecithin to the sodium deoxycholate or the proportion of the soybean lecithin to the polysorbate 80 are both 8.7:1.3, the proportion of the hydrophobic modified polypeptide DP7-C is 55%, the final concentration of the acetyl hexapeptide-8 is 50ppm, the hydration medium is water, and the hydration time is 30 minutes.

The specific preparation process comprises the following steps: accurately weighing soybean lecithin, sodium deoxycholate or polysorbate 80 and vitamin E according to the formula, dissolving with a mixture of chloroform and ethanol at a volume ratio of 1: 2, respectively, dissolving and mixing in a 250ml pear-shaped bottle, performing rotary evaporation at room temperature for 2 hours, and placing the obtained film in a vacuum drying oven overnight. Adding distilled water and a proper amount of hydrophobic modified polypeptide DP7-C in the next day, and performing ultrasonic treatment for 30min by a 400w probe to obtain a blank carrier solution. Slowly adding acetyl hexapeptide-8 to the final concentration of 50ppm, incubating at room temperature for 30 minutes, filtering the obtained mother liquor for multiple times by using a 0.2-micron polycarbonate membrane, adding an excipient of 5% mannitol, subpackaging in penicillin bottles, and freeze-drying to obtain flexible liposome 1-1 containing acetyl hexapeptide-8 and taking sodium deoxycholate as a surfactant and flexible liposome 1-2 containing acetyl hexapeptide-8 and taking polysorbate 80 as a surfactant, wherein the particle diameters of the flexible liposome 1-1 and the flexible liposome 1-2 are respectively about 74.47nm and 68.57nm, as shown in table 1, and the encapsulation rates of the product are respectively 45.6% and 52.65% by an ultracentrifugation method. Wherein, the particle size diagram of the flexible liposome modified by DP7-C and polysorbate 80 as surfactant is shown in figure 1.

Experimental examples 1-2 preparation of PAL-DP7 modified Flexible liposomes comprising acetyl hexapeptide-8

Experimental materials:

soybean lecithin: purchased from Lipoid GmbH;

polysorbate 80: from Merck, USA;

hydrophobically modified polypeptide PAL-DP 7: the Chengdayaki biological medicine science and technology development company is synthesized;

In this experimental example, the flexible liposome is modified by PAL-DP7, the surface active substances are respectively prepared by sodium deoxycholate and polysorbate 80, and the parameters of the preparation process of the flexible liposome containing acetyl hexapeptide-8 are as follows: the ratio of soybean lecithin to sodium deoxycholate or the ratio of soybean lecithin to polysorbate 80 is 8.5:1.5, the ratio of the hydrophobic modified polypeptide PAL-DP7 is 6%, the final concentration of acetyl hexapeptide-8 is 50ppm, the hydration medium is water, and the hydration time is 30 minutes.

The specific preparation process comprises the following steps: accurately weighing soybean lecithin, sodium deoxycholate or polysorbate 80 vitamin E according to the formula, dissolving with a mixture of chloroform and ethanol according to a volume ratio of 1: 2, respectively, dissolving and mixing in a 250ml pear-shaped bottle, performing rotary evaporation at room temperature for 2 hours, and placing the obtained film in a vacuum drying oven overnight. Distilled water and a proper amount of hydrophobic modified polypeptide PAL-DP7 are added in the next day, and a 400w probe is used for ultrasonic treatment for 30min to obtain a blank carrier solution. Slowly adding acetyl hexapeptide-8 to a final concentration of 50ppm, incubating at room temperature for 30 minutes, filtering the obtained mother liquor for multiple times by using a 0.2-micron polycarbonate membrane, adding an excipient of 5% mannitol, subpackaging in a penicillin bottle, and freeze-drying to obtain flexible liposomes 1-3 containing acetyl hexapeptide-8 and taking sodium deoxycholate as surfactants and flexible liposomes 1-4 containing acetyl hexapeptide-8 and taking polysorbate 80 as surfactants, wherein the particle diameters of the flexible liposomes are respectively about 70.34nm and about 59.23nm, as shown in table 1, and the encapsulation rates of the product are respectively 47.2% and 55.17% by an ultracentrifugation method. The flexible liposome modified by PAL-DP7 and using polysorbate 80 as surfactant has the smallest nanometer particle size and the best encapsulation efficiency, and the particle size diagram is shown in figure 2.

TABLE 1 Nanocharacterization of preparation of Flexible liposomes of different composition containing acetyl hexapeptide-8

Experimental example 2 preparation of Flexible Liposome containing palmitoyl tetrapeptide-7

According to the research thought, the palm tetrapeptide-7 flexible liposome is prepared at the same time.

Experimental example 2-1 preparation of DP7-C modified Flexible Liposome containing palmitoyl tetrapeptide-7

Experimental materials:

soybean lecithin: purchased from Lipoid GmbH;

polysorbate 80: from Merck, USA;

palmitoyl tetrapeptide-7: the Chengdayaki biological medicine science and technology development company is synthesized;

In the experimental example, the flexible liposome is modified by DP7-C, the surface active substances are respectively prepared by sodium deoxycholate and polysorbate 80, and the parameters of the preparation process of the flexible liposome containing palmitoyl tetrapeptide-7 are as follows: the ratio of soybean lecithin to sodium deoxycholate or the ratio of soybean lecithin to polysorbate 80 is 8:2, the ratio of the hydrophobic modified polypeptide DP7-C is 5%, the final concentration of the palmitoyl tetrapeptide-7 is 5ppm, the hydration medium is water, and the hydration time is 30 minutes.

The specific preparation process comprises the following steps: accurately weighing soybean lecithin, sodium deoxycholate or polysorbate 80 and vitamin E according to the formula, dissolving with a mixture of chloroform and ethanol at a volume ratio of 1: 2, respectively, dissolving and mixing in a 250ml pear-shaped bottle, performing rotary evaporation at room temperature for 2 hours, and placing the obtained film in a vacuum drying oven overnight. Adding distilled water and a proper amount of hydrophobic modified polypeptide DP7-C in the next day, and performing ultrasonic treatment for 30min by a 400w probe to obtain a blank carrier solution. Slowly adding palmitoyl tetrapeptide-7 to a final concentration of 5ppm, incubating at room temperature for 30 minutes, filtering the obtained mother liquor with a 0.2-micron polycarbonate membrane for multiple times, adding an excipient of 5% mannitol, subpackaging in penicillin bottles, and freeze-drying to obtain the palmitoyl tetrapeptide-7-containing flexible liposome 2-1 taking sodium deoxycholate as a surfactant and the palmitoyl tetrapeptide-7-containing flexible liposome 2-2 taking polysorbate 80 as a surfactant, wherein the particle diameters of the palmitoyl tetrapeptide-7-containing flexible liposome 2-1 and the palmitoyl tetrapeptide-7-containing flexible liposome 2-2 are respectively about 82.69nm and 79.47nm, as shown in Table 2, and the encapsulation efficiencies of the products are respectively 47.65% and 48.1% by ultracentrifugation. Wherein, the particle size diagram of the flexible liposome modified by DP7-C and taking polysorbate 80 as a surfactant is shown in figure 3.

Experimental example 2-2 preparation of PAL-DP7 modified Flexible liposomes comprising palmitoyl tetrapeptide-7

Experimental materials:

soybean lecithin: purchased from Lipoid GmbH;

polysorbate 80: from Merck, USA;

In the experimental example, the flexible liposome is modified by PAL-DP7, the surface active substances are respectively prepared by sodium deoxycholate and polysorbate 80, and the parameters of the preparation process of the flexible liposome containing palmitoyl tetrapeptide-7 are as follows: the ratio of soybean lecithin to sodium deoxycholate or the ratio of soybean lecithin to polysorbate 80 is 8.5:1.5, the ratio of the hydrophobic modified polypeptide PAL-DP7 is 6%, the final concentration of palmitoyl tetrapeptide-7 is 5ppm, the hydration medium is water, and the hydration time is 30 minutes.

The specific preparation process comprises the following steps: accurately weighing soybean lecithin, sodium deoxycholate or polysorbate 80 and vitamin E according to the formula, dissolving with a mixture of chloroform and ethanol at a volume ratio of 1: 2, respectively, dissolving and mixing in a 250ml pear-shaped bottle, performing rotary evaporation at room temperature for 2 hours, and placing the obtained film in a vacuum drying oven overnight. Distilled water and a proper amount of hydrophobic modified polypeptide PAL-DP7 are added in the next day, and a 400w probe is used for ultrasonic treatment for 30min to obtain a blank carrier solution. Slowly adding palmitoyl tetrapeptide-7 to a final concentration of 5ppm, incubating at room temperature for 30 minutes, filtering the obtained mother liquor with a 0.2-micron polycarbonate membrane for multiple times, adding excipient 5% mannitol, subpackaging in penicillin bottles, and lyophilizing to obtain flexible liposome 2-3 containing palmitoyl tetrapeptide-7 and sodium deoxycholate as surfactants and flexible liposome 2-4 containing palmitoyl tetrapeptide-7 and polysorbate 80 as surfactants, wherein the particle diameters are about 75.07nm and about 72.57nm, respectively, as shown in Table 2, and the encapsulation efficiencies of the products are 48.1% and 49.27% respectively, measured by ultracentrifugation. Wherein the flexible liposome modified by PAL-DP7 and with the smallest nanometer particle size and the best encapsulation efficiency is polysorbate 80 as surfactant, and the particle size diagram is shown in figure 4.

TABLE 2 Nanocharacterization of preparation of different composition flexible liposomes containing palmitoyl tetrapeptide-7

Experimental example 3 preparation of Flexible Liposome containing palmitoyl pentapeptide-4

According to the research thought, the palm pentapeptide-4 flexible liposome is prepared at the same time.

Experimental example 3-1 preparation of DP7-C modified Flexible Liposome containing palmitoyl pentapeptide-4

Experimental materials:

soybean lecithin: purchased from Lipoid GmbH;

polysorbate 80: from Merck, USA;

In the experimental example, the flexible liposome is modified by DP7-C, the surface active substances are respectively prepared by sodium deoxycholate and polysorbate 80, and the parameters of the preparation process of the flexible liposome containing palmitoyl pentapeptide-4 are as follows: the proportion of the soybean lecithin to the sodium deoxycholate or the proportion of the soybean lecithin to the polysorbate 80 are both 8.5:1.5, the proportion of the hydrophobic modified polypeptide DP7-C is 5%, the final concentration of the palmitoyl pentapeptide-4 is 10ppm, the hydration medium is water, and the hydration time is 30 minutes.

The specific preparation process comprises the following steps: accurately weighing soybean lecithin, sodium deoxycholate or polysorbate 80 and vitamin E according to the formula, respectively dissolving with ethanol, mixing in 250ml pear-shaped bottle, rotary evaporating at room temperature for 2hr, and placing the obtained film in a vacuum drying oven overnight. Adding distilled water and a proper amount of hydrophobic modified polypeptide DP7-C in the next day, and performing ultrasonic treatment for 30min by a 400w probe to obtain a blank carrier solution. Slowly adding palmitoyl pentapeptide-4 to a final concentration of 10ppm, incubating at room temperature for 30 minutes, filtering the obtained mother liquor with a 0.2-micron polycarbonate membrane for multiple times, adding excipient 5% mannitol, subpackaging in penicillin bottles, and lyophilizing to obtain palmitoyl pentapeptide-4-containing flexible liposome 3-1 using sodium deoxycholate as a surfactant and palmitoyl pentapeptide-4-containing flexible liposome 3-2 using polysorbate 80 as a surfactant, wherein the particle diameters of the palmitoyl pentapeptide-4-containing flexible liposome 3-1 and the flexible liposome 3-2 are respectively 89.37nm and 82.28nm, as shown in Table 3, and the encapsulation efficiencies of the products are respectively 44.3% and 47.12% by ultracentrifugation. Wherein, the particle size diagram of the flexible liposome modified by DP7-C and taking polysorbate 80 as a surfactant is shown in figure 5.

Experimental example 3-2 preparation of PAL-DP7 modified Flexible liposomes comprising palmitoyl pentapeptide-4

Experimental materials:

soybean lecithin: purchased from Lipoid GmbH;

polysorbate 80: from Merck, USA;

In the experimental example, the flexible liposome is modified by PAL-DP7, the surface active substances are respectively prepared by sodium deoxycholate and polysorbate 80, and the parameters of the preparation process of the flexible liposome containing palmitoyl pentapeptide-4 are as follows: the ratio of soybean lecithin to sodium deoxycholate or the ratio of soybean lecithin to polysorbate 80 is 8:2, the ratio of the hydrophobic modified polypeptide PAL-DP7 is 6%, the final concentration of palmitoyl pentapeptide-4 is 10ppm, the hydration medium is water, and the hydration time is 30 minutes.

The specific preparation process comprises the following steps: accurately weighing soybean lecithin, sodium deoxycholate or polysorbate 80 and vitamin E according to the formula, respectively dissolving with ethanol, mixing in 250ml pear-shaped bottle, rotary evaporating at room temperature for 2hr, and placing the obtained film in a vacuum drying oven overnight. Distilled water and a proper amount of hydrophobic modified polypeptide PAL-DP7 are added in the next day, and a 400w probe is used for ultrasonic treatment for 30min to obtain a blank carrier solution. Slowly adding palmitoyl pentapeptide-4 to a final concentration of 10ppm, incubating at room temperature for 30 minutes, filtering the obtained mother liquor with a 0.2-micron polycarbonate membrane for multiple times, adding excipient 5% mannitol, subpackaging in penicillin bottles, and lyophilizing to obtain palmitoyl pentapeptide-4-containing flexible liposome 3-3 using sodium deoxycholate as a surfactant and palmitoyl pentapeptide-4-containing flexible liposome 3-4 using polysorbate 80 as a surfactant, wherein the particle diameters of the palmitoyl pentapeptide-4-containing flexible liposome 3-3 and the palmitoyl pentapeptide-4-containing flexible liposome 3-4 are respectively about 87.24nm and 79.47nm, as shown in Table 3, and the encapsulation efficiencies of the products are respectively 45.1% and 48.72% by ultracentrifugation. Wherein the flexible liposome modified by PAL-DP7 and with the smallest nanometer particle size and the best encapsulation efficiency is polysorbate 80 as surfactant, and the particle size diagram is shown in figure 6.

TABLE 3 Nanocharacterization of preparation of different composition flexible liposomes containing palmitoyl pentapeptide-4

Experimental example 4 preparation of Flexible liposomes containing nonapeptide-1

According to the research thought, the nonapeptide-1 flexible liposome is prepared at the same time.

Experimental example 4-1 preparation of DP7-C modified Flexible liposomes containing nonapeptide-1

Experimental materials:

soybean lecithin: purchased from Lipoid GmbH;

polysorbate 80: from Merck, USA;

nonapeptide-1: the Chengdayaki biological medicine science and technology development company is synthesized;

In the experimental example, the flexible liposome is modified by DP7-C, the surface active substances are respectively prepared by sodium deoxycholate and polysorbate 80, and the parameters of the preparation process of the flexible liposome containing the nonapeptide-1 are as follows: the proportion of the soybean lecithin to the sodium deoxycholate or the proportion of the soybean lecithin to the polysorbate 80 are both 8.2:1.8, the proportion of the hydrophobic modified polypeptide DP7-C is 5%, the final concentration of the nonapeptide-1 is 50ppm, the hydration medium is water, and the hydration time is 30 minutes.

The specific preparation process comprises the following steps: accurately weighing soybean lecithin, sodium deoxycholate or polysorbate 80 and vitamin E according to the formula, dissolving with a mixture of chloroform and ethanol at a volume ratio of 1: 2, respectively, dissolving and mixing in a 250ml pear-shaped bottle, performing rotary evaporation at room temperature for 2 hours, and placing the obtained film in a vacuum drying oven overnight. Adding distilled water and a proper amount of hydrophobic modified polypeptide DP7-C in the next day, and performing ultrasonic treatment for 30min by a 400w probe to obtain a blank carrier solution. Adding nonapeptide-1 slowly to the final concentration of 50ppm, incubating at room temperature for 30min, filtering the obtained mother liquor with 0.2 μm polycarbonate membrane for multiple times, adding excipient 5% mannitol, subpackaging in penicillin bottles, and lyophilizing to obtain nonapeptide-1-containing flexible liposome 4-1 with sodium deoxycholate as surfactant and nonapeptide-1-containing flexible liposome 4-2 with polysorbate 80 as surfactant, wherein the particle diameters are about 69.26nm and 64.94nm, respectively, as shown in Table 4, and the encapsulation efficiencies of the product are 52.3% and 57.26% respectively by ultracentrifugation. Wherein, the particle size diagram of the flexible liposome modified by DP7-C and taking polysorbate 80 as a surfactant is shown in figure 7.

Experimental example 4-2 preparation of PAL-DP7 modified Flexible liposomes comprising nonapeptide-1

Experimental materials:

soybean lecithin: purchased from Lipoid GmbH;

polysorbate 80: from Merck, USA;

In this experimental example, the flexible liposome is modified by PAL-DP7, the surface active substances are respectively prepared by sodium deoxycholate and polysorbate 80, and the parameters of the preparation process of the flexible liposome containing nonapeptide-1 are as follows: the ratio of soybean lecithin to sodium deoxycholate or the ratio of soybean lecithin to polysorbate 80 is 8:2, the ratio of the hydrophobic modified polypeptide PAL-DP7 is 6%, the final concentration of nonapeptide-1 is 50ppm, the hydration medium is water, and the hydration time is 30 minutes.

The specific preparation process comprises the following steps: accurately weighing soybean lecithin, sodium deoxycholate or polysorbate 80 and vitamin E according to the formula, dissolving with a mixture of chloroform and ethanol at a volume ratio of 1: 2, respectively, dissolving and mixing in a 250ml pear-shaped bottle, performing rotary evaporation at room temperature for 2 hours, and placing the obtained film in a vacuum drying oven overnight. Distilled water and a proper amount of hydrophobic modified polypeptide PAL-DP7 are added in the next day, and a 400w probe is used for ultrasonic treatment for 30min to obtain a blank carrier solution. Adding nonapeptide-1 slowly to the final concentration of 50ppm, incubating at room temperature for 30min, filtering the obtained mother liquor with 0.2 μm polycarbonate membrane for multiple times, adding excipient 5% mannitol, subpackaging in penicillin bottles, and lyophilizing to obtain nonapeptide-1-containing flexible liposome 4-3 with sodium deoxycholate as surfactant and nonapeptide-1-containing flexible liposome 4-4 with polysorbate 80 as surfactant, wherein the particle diameters are 65.4nm and 62.11nm respectively, as shown in Table 4, and the encapsulation efficiencies of the product are 54.1% and 59.34% respectively by ultracentrifugation. Wherein the flexible liposome modified by PAL-DP7 and with the smallest nanometer particle size and the best encapsulation efficiency is polysorbate 80 as surfactant, and the particle size diagram is shown in figure 8.

TABLE 4 Nanocharacterization of preparation of Flexible liposomes of different composition containing nonapeptide-1

Experimental example 5 preparation of Flexible liposomes containing acetyl tetrapeptide-5

According to the research thought, the flexible liposome of acetyl tetrapeptide-5 is prepared at the same time.

Experimental example 5-1 preparation of DP7-C modified Flexible Liposome containing acetyl tetrapeptide-5

Experimental materials:

soybean lecithin: purchased from Lipoid GmbH;

polysorbate 80: from Merck, USA;

acetyl tetrapeptide-5: the Chengdayaki biological medicine science and technology development company is synthesized;

In the experimental example, the flexible liposome is modified by DP7-C, the surface active substances are respectively prepared by sodium deoxycholate and polysorbate 80, and the preparation process of the flexible liposome containing acetyl tetrapeptide-5 has the following parameters: the proportion of the soybean lecithin to the sodium deoxycholate or the proportion of the soybean lecithin to the polysorbate 80 are both 8.5:1.5, the proportion of the hydrophobic modified polypeptide DP7-C is 5%, the final concentration of the acetyl tetrapeptide-5 is 100ppm, the hydration medium is water, and the hydration time is 30 minutes.

The specific preparation process comprises the following steps: accurately weighing soybean lecithin, sodium deoxycholate or polysorbate 80 and vitamin E according to the formula, dissolving with a mixture of chloroform and ethanol at a volume ratio of 1: 3, respectively, dissolving and mixing in a 250ml pear-shaped bottle, performing rotary evaporation at room temperature for 2 hours, and placing the obtained film in a vacuum drying oven overnight. Adding distilled water and a proper amount of hydrophobic modified polypeptide DP7-C in the next day, and performing ultrasonic treatment for 30min by a 400w probe to obtain a blank carrier solution. Slowly adding acetyl tetrapeptide-5 to the final concentration of 100ppm, incubating at room temperature for 30 minutes, filtering the obtained mother liquor with 0.2 μm polycarbonate membrane for multiple times, adding excipient 5% mannitol, subpackaging in penicillin bottles, and lyophilizing to obtain flexible liposome 5-1 containing acetyl tetrapeptide-5 with sodium deoxycholate as surfactant and flexible liposome 5-2 containing acetyl tetrapeptide-5 with polysorbate 80 as surfactant, wherein the particle diameters are respectively 77.48nm and 64.52nm, as shown in Table 4, and the encapsulation efficiencies of the product are respectively 51.4% and 53.6% by ultracentrifugation. Wherein, the particle size diagram of the flexible liposome modified by DP7-C and taking polysorbate 80 as a surfactant is shown in figure 9.

Experimental example 5-2 preparation of PAL-DP7 modified Flexible liposomes containing acetyl tetrapeptide-5

Experimental materials:

soybean lecithin: purchased from Lipoid GmbH;

polysorbate 80: from Merck, USA;

In this experimental example, the flexible liposome was modified with PAL-DP7, the surface active substances were prepared with sodium deoxycholate and polysorbate 80, respectively, and the preparation process of the flexible liposome containing acetyl tetrapeptide-5 was carried out with the following parameters: the ratio of soybean lecithin to sodium deoxycholate or the ratio of soybean lecithin to polysorbate 80 is 8:2, the ratio of the hydrophobic modified polypeptide PAL-DP7 is 6%, the final concentration of acetyl tetrapeptide-5 is 100ppm, the hydration medium is water, and the hydration time is 30 minutes.

The specific preparation process comprises the following steps: accurately weighing soybean lecithin, sodium deoxycholate or polysorbate 80 and vitamin E according to the formula, dissolving with a mixture of chloroform and ethanol at a volume ratio of 1: 3, respectively, dissolving and mixing in a 250ml pear-shaped bottle, performing rotary evaporation at room temperature for 2 hours, and placing the obtained film in a vacuum drying oven overnight. Distilled water and a proper amount of hydrophobic modified polypeptide PAL-DP7 are added in the next day, and a 400w probe is used for ultrasonic treatment for 30min to obtain a blank carrier solution. Slowly adding acetyl tetrapeptide-5 to the final concentration of 100ppm, incubating at room temperature for 30 minutes, filtering the obtained mother liquor with 0.2 μm polycarbonate membrane for multiple times, adding excipient 5% mannitol, subpackaging in penicillin bottles, and lyophilizing to obtain flexible liposome 5-3 containing acetyl tetrapeptide-5 and polysorbate 80 as surfactants, wherein the particle diameters of the flexible liposome 5-4 are respectively 66.9nm and 64.12nm, as shown in Table 5, and the encapsulation efficiencies of the product are respectively 53.2% and 56.5% by ultracentrifugation. Wherein the flexible liposome modified by PAL-DP7 and with the smallest nanometer particle size and the best encapsulation efficiency is polysorbate 80 as surfactant, and the particle size diagram is shown in figure 10.

TABLE 5 Nanocharacterization of preparation of Flexible liposomes of different composition containing acetyl tetrapeptide-5

Experimental example 6 preparation of Flexible liposomes containing myristoyl pentapeptide-17

According to the research thought, the myristoyl pentapeptide-17 flexible liposome is prepared at the same time.

Experimental example 6-1 preparation of DP7-C modified Flexible Liposome containing Myristicamyl pentapeptide-17

Experimental materials:

soybean lecithin: purchased from Lipoid GmbH;

polysorbate 80: from Merck, USA;

myristoyl pentapeptide-17: the Chengdayaki biological medicine science and technology development company is synthesized;

In the experimental example, the flexible liposome is modified by DP7-C, the surface active substances are respectively prepared by sodium deoxycholate and polysorbate 80, and the preparation process of the flexible liposome containing the myristoyl pentapeptide-17 has the following parameters: the proportion of the soybean lecithin to the sodium deoxycholate or the proportion of the soybean lecithin to the polysorbate 80 are both 8.5:1.5, the proportion of the hydrophobic modified polypeptide DP7-C is 5%, the final concentration of the myristoyl pentapeptide-17 is 200ppm, the hydration medium is water, and the hydration time is 30 minutes.

The specific preparation process comprises the following steps: accurately weighing soybean lecithin, sodium deoxycholate or polysorbate 80 and vitamin E according to the formula, dissolving with a mixture of chloroform and ethanol according to a volume ratio of 1: 1, dissolving and mixing in a 250ml pear-shaped bottle, performing rotary evaporation at room temperature for 2 hours, and placing the obtained film in a vacuum drying oven overnight. Adding distilled water and a proper amount of hydrophobic modified polypeptide DP7-C in the next day, and performing ultrasonic treatment for 30min by a 400w probe to obtain a blank carrier solution. Slowly adding the myristoyl pentapeptide-17 to the final concentration of 200ppm, incubating for 30 minutes at room temperature, filtering the obtained mother liquor for multiple times by using a 0.2 mu m polycarbonate membrane, adding an excipient of 5% mannitol, subpackaging in penicillin bottles, and freeze-drying to obtain the flexible liposome 6-1 containing the myristoyl pentapeptide-17 and taking sodium deoxycholate as a surfactant and the flexible liposome 6-2 containing the myristoyl pentapeptide-17 and taking polysorbate 80 as a surfactant, wherein the particle diameters of the flexible liposome 6-1 and the flexible liposome 6-2 are respectively about 131.4nm and 120.4nm, as shown in Table 4, and the encapsulation efficiencies of the product are respectively 47.4% and 51.16% by an ultracentrifugation method. Wherein, the particle size diagram of the flexible liposome modified by DP7-C and taking polysorbate 80 as a surfactant is shown in figure 11.

Experimental example 6-2 preparation of PAL-DP7 modified Flexible liposomes comprising myristoyl pentapeptide-17

Experimental materials:

soybean lecithin: purchased from Lipoid GmbH;

polysorbate 80: from Merck, USA;

In the experimental example, the flexible liposome is modified by PAL-DP7, the surface active substances are respectively prepared by sodium deoxycholate and polysorbate 80, and the preparation process of the flexible liposome containing myristoyl pentapeptide-17 has the following parameters: the ratio of soybean lecithin to sodium deoxycholate or the ratio of soybean lecithin to polysorbate 80 is 8:2, the ratio of the hydrophobic modified polypeptide PAL-DP7 is 6%, the final concentration of myristoyl pentapeptide-17 is 200ppm, the hydration medium is water, and the hydration time is 30 minutes.

The specific preparation process comprises the following steps: accurately weighing soybean lecithin, sodium deoxycholate or polysorbate 80 and vitamin E according to the formula, dissolving with a mixture of chloroform and ethanol according to a volume ratio of 1: 1, dissolving and mixing in a 250ml pear-shaped bottle, performing rotary evaporation at room temperature for 2 hours, and placing the obtained film in a vacuum drying oven overnight. Distilled water and a proper amount of hydrophobic modified polypeptide PAL-DP7 are added in the next day, and a 400w probe is used for ultrasonic treatment for 30min to obtain a blank carrier solution. Slowly adding the myristoyl pentapeptide-17 to the final concentration of 200ppm, incubating for 30 minutes at room temperature, filtering the obtained mother liquor for multiple times by using a 0.2 mu m polycarbonate membrane, adding an excipient of 5% mannitol, subpackaging in penicillin bottles, and freeze-drying to obtain the flexible liposome 6-3 containing the myristoyl pentapeptide-17 and taking sodium deoxycholate as a surfactant and the flexible liposome 6-4 containing the myristoyl pentapeptide-17 and taking polysorbate 80 as a surfactant, wherein the particle diameters of the flexible liposome 6-3 and the flexible liposome 6-4 are respectively about 121.4nm and 114.1nm, as shown in Table 6, and the encapsulation efficiencies of the product are respectively 49.2% and 54.1% by adopting an ultracentrifugation method. Wherein the flexible liposome modified by PAL-DP7 and with the smallest nanometer particle size and the best encapsulation efficiency is polysorbate 80 as surfactant, and the particle size diagram is shown in figure 12.

TABLE 6 Nanocharacterization of preparation of flexible liposomes of different compositions containing myristoyl pentapeptide-17

Example 2 transdermal effect verification of Flexible liposomes containing active Polypeptides of the invention

In vitro transdermal experiments, an improved single-chamber Franz diffusion cell is adopted, mouse skin is used as the skin of the in vitro transdermal experiment, the transdermal absorption results of different flexible liposomes containing active polypeptide are compared, sampling is carried out at different time points respectively, the concentration of the drug in the transdermal receiving solution is measured by an HPLC method, and the accumulated transdermal quantity is calculated.

Experimental example 2-1 verification of transdermal Effect of Flexible liposomes comprising acetyl hexapeptide-8

1. Material

Sample preparation: experimental example 1 prepared acetyl hexapeptide-8-containing liposomes lyophilized powder (1-1, 1-2, 1-3, 1-4) modified with polysorbate 80 as a surfactant and PAL-DP7, and acetyl hexapeptide-8-containing liposomes unmodified with DP7-C or PAL-DP7 were used as a common liposomes control;

the instrument comprises the following steps: a drug transdermal test diffusion instrument (Shanghai yellow sea drug test, model RYJ-6B); HPLC (Agilent, model 1260); a chromatographic column: ZORBAX 300 SB-C18;

materials: stripped mouse skin.

2. Method of producing a composite material

2.1 the transdermal test of the sample is carried out according to the instructions of the diffusion instrument of the transdermal test, and a blank control with buffer solution added above is arranged at the same time, and the samples in the receiving pool are harvested respectively at 0hr, 1hr, 2hr, 4hr and 8hr of the test.

2.2 HPLC detection: ultraviolet detection wavelength: 220 nm; mobile phase: solution A: 100% ACN + 0.1% TFA; and B, liquid B: 100% H₂O + 0.1% TFA; the test conditions are shown in table 7 below:

TABLE 7 HPLC DETECTION CONDITIONS TABLE

Time (min)	Solution A	Liquid B
				0	43％	57％
20	63％	37％
			20.5	43％	57％
27	43％	57％

3. Results

Respectively taking the polypeptide of the acetyl hexapeptide-8 as a standard substance and a sample for HPLC determination, establishing a standard curve according to the peak area of the acetyl hexapeptide-8 standard substance, substituting the peak area of the sample to be determined into a formula, and calculating the cumulative transdermal mass of the sample, as shown in figure 13. The results show that the amount of acetyl hexapeptide-8 penetrating the skin of mice increases gradually with time, and that DP7-C and PAL-DP7 modified flexible liposomes have better transdermal properties than unmodified flexible liposomes.

Experimental example 2-2 verification of transdermal Effect of Flexible liposomes containing palmitoyl tetrapeptide-7

1. Material

Sample preparation: experimental example 2 prepared palmitoyl tetrapeptide-7-containing flexo liposome lyophilized powder (2-1, 2-2, 2-3, 2-4) modified with polysorbate 80 as a surfactant and PAL-DP7, and palmitoyl tetrapeptide-7-containing flexo liposome without DP7-C or PAL-DP7 modification were used as a common flexo liposome control;

materials: stripped mouse skin.

2. Method of producing a composite material

2.2 HPLC detection: ultraviolet detection wavelength: 220 nm; mobile phase: solution A: 100% ACN + 0.1% TFA; and B, liquid B: 100% H₂O + 0.1% TFA; the test conditions are shown in Table 8 below:

TABLE 8 HPLC DETECTION CONDITIONS TABLE

Time (min)	Solution A	Liquid B
				0	50％	50％
25	75％	25％
			25.01	100％	0
30	100％	0
			30.01	50％	50％
35	50％	50％

3. Results

Respectively taking the polypeptide of the palmitoyl tetrapeptide-7 as a standard substance and a sample for HPLC determination, establishing a standard curve according to the peak area of the palmitoyl tetrapeptide-7 standard substance, substituting the peak area of the sample to be determined into a formula, and calculating the cumulative transdermal mass of the sample, as shown in figure 14. The results show that the amount of palmitoyl tetrapeptide-7 penetrating the skin of mice increases gradually with time, and that both DP7-C and PAL-DP7 modified flexible liposomes have better transdermal properties compared to unmodified flexible liposomes.

Experimental examples 2-3 verification of transdermal Effect of Flexible liposomes containing palmitoyl pentapeptide-4

1. Material

Sample preparation: experimental example 3 prepared palmitoyl pentapeptide-4-containing flexo liposome lyophilized powder (3-1, 3-2, 3-3, 3-4) modified with polysorbate 80 as a surfactant and PAL-DP7, and palmitoyl pentapeptide-4-containing flexo liposome without DP7-C or PAL-DP7 modification were used as common flexo liposome controls;

materials: stripped mouse skin.

2. Method of producing a composite material

2.2 HPLC detection: ultraviolet detection wavelength: 220 nm; mobile phase: solution A: 100% ACN + 0.1% TFA; and B, liquid B: 100% H₂O + 0.1% TFA; the test conditions are shown in table 9 below:

TABLE 9 HPLC DETECTION CONDITIONS TABLE

3. Results

Respectively taking the polypeptide of the palmitoyl pentapeptide-4 as a standard substance and a sample to carry out HPLC (high performance liquid chromatography) determination, establishing a standard curve according to the peak area of the palmitoyl pentapeptide-4 standard substance, substituting the peak area of the sample to be determined into a formula, and calculating the cumulative transdermal mass of the sample, as shown in figure 15. The results show that the amount of palmitoyl pentapeptide-4 penetrating the skin of mice increases gradually with time, and that both DP7-C and PAL-DP7 modified flexible liposomes have better transdermal properties compared to unmodified flexible liposomes.

Experimental examples 2-4 verification of transdermal Effect of Flexible liposomes containing nonapeptide-1

1. Material

Sample preparation: experimental example 4 prepared nonapeptide-1-containing flexo liposome lyophilized powder (4-1, 4-2, 4-3, 4-4) modified with polysorbate 80 as a surfactant and PAL-DP7, and nonapeptide-1-containing flexo liposomes that were not modified with DP7-C or PAL-DP7 were used as a common flexo liposome control;

materials: stripped mouse skin.

2. Method of producing a composite material

2.2 HPLC detection: ultraviolet detection wavelength: 220 nm; mobile phase: solution A: 100% ACN + 0.1% TFA; and B, liquid B: 100% H₂O + 0.1% TFA; the test conditions are shown in table 10 below:

TABLE 10 HPLC DETECTION CONDITIONS TABLE

3. Results

Respectively taking the polypeptide of the nonapeptide-1 as a standard substance and a sample for HPLC determination, establishing a standard curve according to the peak area of the nonapeptide-1 standard substance, substituting the peak area of the sample to be determined into a formula, and calculating the cumulative transdermal mass of the sample, as shown in figure 16. The results show that the amount of nonapeptide-1 penetrating the skin of mice increases gradually with time, and that DP7-C and PAL-DP7 modified flexible liposomes have better transdermal properties than unmodified flexible liposomes.

Experimental examples 2-5 verification of transdermal Effect of Flexible liposomes comprising acetyl tetrapeptide-5

1. Material

Sample preparation: experimental example 5 prepared acetyl tetrapeptide-5-containing flexo liposome lyophilized powder (5-1, 5-2, 5-3, 5-4) modified with polysorbate 80 as a surfactant and PAL-DP7, and acetyl tetrapeptide-5-containing flexo liposome without DP7-C or PAL-DP7 modification were used as a general flexo liposome control;

materials: stripped mouse skin.

2. Method of producing a composite material

2.2 HPLC detection: ultraviolet detection wavelength: 220 nm; mobile phase: solution A: 100% ACN + 0.1% TFA; and B, liquid B: 100% H₂O + 0.1% TFA; the test conditions are shown in Table 11 below:

TABLE 11 HPLC TEST CONDITIONS TABLE

3. Results

Respectively taking the polypeptide of acetyl tetrapeptide-5 as a standard substance and a sample for HPLC determination, establishing a standard curve according to the peak area of the acetyl tetrapeptide-5 standard substance, substituting the peak area of the sample to be determined into a formula, and calculating the cumulative transdermal mass of the sample, as shown in figure 17. The results show that the amount of acetyl tetrapeptide-5 penetrating the skin of mice increases gradually with time, and that DP7-C and PAL-DP7 modified flexible liposomes have better transdermal properties than unmodified flexible liposomes.

Test examples 2-6 verification of transdermal Effect of Flexible liposomes containing myristoyl pentapeptide-17

1. Material

Sample preparation: experimental example 6 prepared myristoyl pentapeptide-17-containing flexo liposome lyophilized powder (6-1, 6-2, 6-3, 6-4) modified with polysorbate 80 as a surfactant and PAL-DP7, and myristoyl pentapeptide-17-containing flexo liposome without DP7-C or PAL-DP7 modification were used as a common flexo liposome control;

materials: stripped mouse skin.

2. Method of producing a composite material

2.2 HPLC detection: ultraviolet detection wavelength: 220 nm; mobile phase: solution A: 100% ACN + 0.1% TFA; and B, liquid B: 100% H₂O + 0.1% TFA; the test conditions are shown in table 12 below:

TABLE 12 HPLC TEST CONDITIONS TABLE

3. Results

Respectively taking the polypeptide of the myristyl pentapeptide-4 as a standard substance and a sample for HPLC determination, establishing a standard curve according to the peak area of the myristyl pentapeptide-4 standard substance, substituting the peak area of the sample to be determined into a formula, and calculating the cumulative transdermal volume of the sample, as shown in figure 18. The results show that the amount of acetyl pentapeptide-4 penetrating the skin of mice increases gradually with time, and that DP7-C and PAL-DP7 modified flexible liposomes have better transdermal properties than unmodified flexible liposomes.

Example 3 stability study of Flexible Liposome lyophilized powder containing active polypeptide of the invention

The flexible liposome freeze-dried powder containing the active polypeptide prepared in the example 1 is subjected to stability study (comprising three aspects of accelerated test, long-term test and the like). The samples were placed in a 37 ℃ drug stability test chamber for accelerated testing, and were placed for 1, 2, 3, and 6 months, respectively, and the properties of the lyophilized powder, including color, content, and solubility, were determined, with the results shown in table 13. The sample is placed in a drug stability test box at 25 ℃ for long-term test, and is respectively placed for 3, 6, 9, 12, 15, 18, 21 and 24 months, and the color, HPLC content and solubility of the freeze-dried powder are respectively detected, and the result is shown in Table 14.

TABLE 13 stability acceleration test of Flexible Liposome lyophilized powder containing active polypeptide (37 deg.C)

TABLE 14 stability Long-term test (25 ℃ C.) for Flexible Liposome lyophilized powders containing active polypeptide

The results show that the freeze-dried powder keeps the properties and the activity of the sample in the first 3 months of the accelerated test, the activity is reduced in the 6 th month, and the high-temperature storage is avoided as much as possible. The freeze-dried powder keeps better properties and activity in a long-term test at 25 ℃, and can ensure that the activity is not lost after 2 years of storage.

Example 4 preparation of essence of Flexible liposomes containing active Polypeptides according to the invention

The flexible liposome containing the active polypeptide can be prepared into essence by using a solvent for use. The formula of the solvent is as follows: 0.3 percent of hyaluronic acid, 0.1 percent of allantoin, 0.5 percent of collagen, 0.1 percent of EDTA and 0.15 percent of multivitamin. The formula is aseptically subpackaged into 10-30 ml bottled solvents. When in use, 1 bottle of freeze-dried flexible liposome is dissolved by 1 bottle of solvent to obtain the essence which is directly smeared on the face after cleaning the face in the morning and evening.

Test example 4-1 verification of the effect of use of the essence of flexible liposome containing acetyl hexapeptide-8:

subject: 108 volunteers aged 40-60 years;

the experimental method comprises the following steps: volunteers were randomly divided into two groups, one group being the essence of flexible liposomes containing acetyl hexapeptide-8 prepared in example 4, and the other group being the vehicle without flexible liposomes. After cleaning the face in the morning and evening every day, the two groups of essences are respectively applied to the face and gently massaged until the essences are absorbed, and other skin care products are not applied. The trial period is 2-6 weeks, and after trial, a use effect statistical table is filled, and the main indexes comprise functions of skin care, fine line removal and the like.

The experimental results are as follows: the essence is tried for 108 people all over the country, such as Chengdu, Wenchang, Tibet, Chongqing, Wuhan, Hainan, Shenzhen and the like, the trial effect evaluation is better (see table 15 for details), and simultaneously 1 case of allergic symptoms does not appear, which indicates that the essence has good safety.

TABLE 15 serum usage results of flexible liposomes containing acetyl hexapeptide-8 of the present invention

Item	Number of trial persons	Service time (week)	Obvious effect (%)	The effect was not remarkable (%)
					Skin care	108	2-6	82	18
Removing wrinkles	108	4-6	88	12

From the above results, it can be seen that: the flexible liposome containing acetyl hexapeptide-8 can be applied to cosmetics, the transdermal efficiency is high, the effect is good, the prepared cosmetics show good effects in nursing and anti-wrinkle surfaces, and the application prospect is wide.

Test example 4-2 verification of the effect of using the essence of palmitoyl tetrapeptide-7-containing flexible liposome:

subject: 188 volunteers aged 40-60 years;

the experimental method comprises the following steps: volunteers were randomly divided into two groups, one containing the essence of the flexible liposomes containing palmitoyl tetrapeptide-7 prepared in example 4, and the other containing the vehicle without flexible liposomes. After cleaning the face in the morning and evening every day, the two groups of essences are respectively applied to the face and gently massaged until the essences are absorbed, and other skin care products are not applied. The trial period is 2-6 weeks, and after trial, a use effect statistical table is filled, and the main indexes comprise functions of skin care, fine line removal and the like.

The experimental results are as follows: the essence is tried for nearly 200 people all over the country, such as Chengdu, Wenchang, Tibet, Chongqing, Wuhan, Hainan, Shenzhen and the like, the trial effect evaluation is better (see table 16 for details), and simultaneously 1 case of allergic symptoms does not appear, which indicates that the essence has good safety.

TABLE 16 serum usage results of the palmitoyl tetrapeptide-7-containing flexible liposomes of the present invention

From the above results, it can be seen that: the flexible liposome containing palmitoyl tetrapeptide-7 can be applied to cosmetics, has high transdermal efficiency and good effect, and the prepared cosmetics have excellent effects on resisting free radicals and oxidation, removing acnes and freckles, repairing sunburn and resisting allergy, and have wide application prospects.

Experimental example 4-3 verification of the effect of using the essence of palmitoyl pentapeptide-4-containing flexible liposome:

subject: 90 volunteers in the age range of 40-60 years;

the experimental method comprises the following steps: volunteers were randomly divided into two groups, one containing the essence of the flexible liposomes containing palmitoyl pentapeptide-4 prepared in example 4, and the other containing the vehicle without flexible liposomes. After cleaning the face in the morning and evening every day, the two groups of essences are respectively applied to the face and gently massaged until the essences are absorbed, and other skin care products are not applied. The trial period is 2-6 weeks, and after trial, a use effect statistical table is filled, and the main indexes comprise functions of skin care, fine line removal, elasticity, smoothness and the like.

The experimental results are as follows: the essence is tried for nearly 100 people in all parts of the country, such as Chengdu, Wenchang, Tibet, Chongqing, Wuhan, Hainan, Shenzhen and the like, the trial effect evaluation is better (see table 17 in detail), and simultaneously 1 case of allergic symptoms does not appear, which indicates that the essence has good safety.

TABLE 17 serum usage results of the palmitoyl pentapeptide-4-containing flexible liposomes of the present invention

Item	Number of trial persons	Service time (week)	Obvious effect (%)	The effect was not remarkable (%)
					Skin care	90	2-6	92	8
Removing wrinkles	90	4-6	84	16
					Elastic smooth	90	2-6	90	10

From the above results, it can be seen that: the palmitoyl pentapeptide-4-containing flexible liposome can be applied to cosmetics, has high transdermal efficiency and good effect, and the prepared cosmetics have good effects on skin care, wrinkle resistance and elasticity smoothness and have wide application prospects.

Experimental examples 4-4 verification of the effect of using the serum containing nonapeptide-1 flexible liposomes:

subject: 80 volunteers aged 20-40 years;

the experimental method comprises the following steps: volunteers were randomly divided into two groups, one group being the serum containing the nonapeptide-1 flexible liposomes prepared in example 4, and the other group being the vehicle without the flexible liposomes. After cleaning the face in the morning and evening every day, the two groups of essences are respectively applied to the face and gently massaged until the essences are absorbed, and other skin care products are not applied. The trial period is 4-8 weeks, and after trial, a using effect statistical table is filled, and the main indexes comprise functions of skin care, whitening, freckle removal and the like.

The experimental results are as follows: the essence is tried for nearly 100 people in all parts of the country, such as Chengdu, Wenchang, Tibet, Chongqing, Wuhan, Hainan, Shenzhen and the like, the trial effect evaluation is better (see table 18 for details), and simultaneously 1 case of allergic symptoms does not appear, which indicates that the essence has good safety.

TABLE 18 serum usage results of the nonapeptide-1-containing flexible liposomes of the present invention

Item	Number of trial persons	Service time (week)	Obvious effect (%)	EffectNot obvious (%)
					Skin care	80	4-8	85	15
Whitening	80	4-8	88	12
					Speckle removing method	30	4-8	80	20

From the above results, it can be seen that: the nonapeptide-1-containing flexible liposome can be applied to cosmetics, has high transdermal efficiency and good effect, and the prepared cosmetics show good effects in whitening and removing freckles and have wide application prospects.

Sequence listing

<110> Shenzhen Shenshangke scientific and technology Limited

<120> flexible liposome cosmetic containing active polypeptide and preparation method thereof

<130> A190061K

<141> 2019-01-30

<150> 201810150521.3

<151> 2018-02-13

<160> 2

<170> SIPOSequenceListing 1.0

<210> 1

<211> 12

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 1

Val Gln Trp Arg Ile Arg Val Ala Val Ile Arg Lys

1 5 10

<210> 2

<211> 9

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 2

Met Pro Phe Arg Trp Phe Lys Pro Val

1 5

Claims

1. Flexible liposome cosmetic products comprising active polypeptides, characterized in that: is made of a hydrophobic modified polypeptide modified flexible liposome containing active polypeptide; the hydrophobic modified polypeptide modified flexible liposome is prepared from the following raw materials in parts by weight: lecithin sodium deoxycholate or polysorbate 80 = 7:2-4, 1-10 wt% of hydrophobic modified polypeptide and active polypeptide, wherein the hydrophobic modified polypeptide is connected with NH at C terminal based on SEQ ID NO:1₂And coupling cholesterol or palmitic acid to the nitrogen terminus of the polypeptide.

2. A flexible liposomal cosmetic comprising an active polypeptide according to claim 1, characterized in that: also contains antioxidant.

3. A flexible liposomal cosmetic comprising an active polypeptide according to claim 2, characterized in that: the antioxidant comprises: at least one of vitamin C, vitamin E or coenzyme Q.

4. A flexible liposomal cosmetic comprising an active polypeptide according to claim 2, characterized in that: the weight percentage of the antioxidant is 0.1-1%.

5. A flexible liposomal cosmetic comprising an active polypeptide according to claim 1, characterized in that: the active polypeptide is an active polypeptide which can be used in the field of cosmetics.

6. Flexible liposomal cosmetic comprising an active polypeptide according to claim 5, characterized in that: the active polypeptide used in the field of cosmetics is an active polypeptide having at least one function of anti-wrinkle, anti-free radical, cell activity promotion, whitening, detumescence or eyelash growth promotion.

7. Flexible liposomal cosmetic comprising an active polypeptide according to claim 6, characterized in that: the active polypeptide comprises at least one of copper peptide, dipeptide-2, palmitoyl tripeptide-1, palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, acetyl tetrapeptide-5, acetyl tetrapeptide-9, palmitoyl pentapeptide-3, palmitoyl pentapeptide-4, acetyl hexapeptide-8, acetyl octapeptide, myristoyl pentapeptide-17, myristoyl hexapeptide-4, nonapeptide-1, carnosine, L-carnosine or glutathione.

8. A flexible liposomal cosmetic comprising an active polypeptide according to claim 1, characterized in that: the weight percentage of the active polypeptide is 0.01-1%.

9. The cosmetic of flexible liposomes containing active polypeptides of claim 1, wherein said flexible liposomes are prepared from the following components in the following proportions: lecithin sodium deoxycholate or polysorbate 80 = 7:3 by weight, 0.5% by weight of antioxidant, and 2-10% by weight of hydrophobically modified polypeptide.

10. A flexible liposomal cosmetic comprising an active polypeptide according to claim 1, characterized in that: the structure of the hydrophobic modified polypeptide is as follows:

，

wherein R is

。

11. Flexible liposomal cosmetic comprising an active polypeptide according to claim 6, characterized in that: the anti-wrinkle active polypeptide is at least one of palmitoyl tripeptide, acetyl hexapeptide-8 or acetyl octapeptide.

12. A flexible liposomal cosmetic comprising an active polypeptide according to claim 11, characterized in that: the anti-wrinkle active polypeptide accounts for 0.01-3% of the liposome by weight.

13. A flexible liposomal cosmetic comprising an active polypeptide according to claim 11, characterized in that: the anti-wrinkle active polypeptide accounts for 0.05-2% of the liposome by weight.

14. A flexible liposomal cosmetic comprising an active polypeptide according to claim 11, characterized in that: the anti-wrinkle active polypeptide accounts for 0.1-1% of the liposome by weight.

15. Flexible liposomal cosmetic comprising an active polypeptide according to claim 6, characterized in that: the anti-free radical active polypeptide is at least one of glutathione, palmitoyl tetrapeptide-7, carnosine or L-carnosine.

16. A flexible liposomal cosmetic comprising an active polypeptide according to claim 15, characterized in that: the anti-free radical active polypeptide accounts for 0.01-3% of the liposome by weight.

17. A flexible liposomal cosmetic comprising an active polypeptide according to claim 15, characterized in that: the anti-free radical active polypeptide accounts for 0.05-2% of the liposome by weight.

18. A flexible liposomal cosmetic comprising an active polypeptide according to claim 15, characterized in that: the anti-free radical active polypeptide accounts for 0.1-1% of the liposome by weight.

19. Flexible liposomal cosmetic comprising an active polypeptide according to claim 6, characterized in that: the active polypeptide for promoting cell activity is at least one of palmitoyl pentapeptide-3, palmitoyl pentapeptide-4, copper peptide, palmitoyl tripeptide-1, palmitoyl tripeptide-5, acetyl tetrapeptide-9 or myristoyl hexapeptide-4.

20. A flexible liposomal cosmetic comprising an active polypeptide according to claim 19, characterized in that: the weight percentage of the cell activity promoting active polypeptide in the liposome is 0.01-3%.

21. A flexible liposomal cosmetic comprising an active polypeptide according to claim 19, characterized in that: the weight percentage of the cell activity promoting active polypeptide in the liposome is 0.05-2%.

22. A flexible liposomal cosmetic comprising an active polypeptide according to claim 19, characterized in that: the weight percentage of the cell activity promoting active polypeptide in the liposome is 0.1-1%.

23. Flexible liposomal cosmetic comprising an active polypeptide according to claim 6, characterized in that: the whitening active polypeptide is nonapeptide-1.

24. Flexible liposomal cosmetic comprising an active polypeptide according to claim 6, characterized in that: the detumescence active polypeptide is acetyl tetrapeptide-5 or dipeptide-2.

25. Flexible liposomal cosmetic comprising an active polypeptide according to claim 6, characterized in that: the active polypeptide for promoting eyelash growth is myristoyl pentapeptide-17.

26. A flexible liposomal cosmetic comprising an active polypeptide according to any one of claims 23-25 characterised in that: the active polypeptide for whitening, detumescence and promoting eyelash growth accounts for 0.01-3% of the liposome by weight.

27. A flexible liposomal cosmetic comprising an active polypeptide according to any one of claims 23-25 characterised in that: the active polypeptide for whitening, detumescence and promoting eyelash growth accounts for 0.05-2% of the liposome by weight.

28. A flexible liposomal cosmetic comprising an active polypeptide according to any one of claims 23-25 characterised in that: the active polypeptide for whitening, detumescence and promoting eyelash growth accounts for 0.1-1% of the liposome by weight.

29. A flexible liposomal cosmetic comprising an active polypeptide according to claim 1, characterized in that: the flexible liposome cosmetic containing active polypeptide also comprises at least one of emulsifier, co-emulsifier, skin conditioner, whitening agent, colorant, humectant, solubilizer, surfactant, antiseptic, aromatic, emollient, anti-acne agent, film forming agent, thickener, pH regulator, buffer, stabilizer or ultraviolet absorbent.

30. A flexible liposomal cosmetic comprising an active polypeptide according to claim 1, characterized in that: the flexible liposome cosmetic containing the active polypeptide is in the dosage form of aqua, emulsion, paste, jelly, troche or aerosol.

31. A flexible liposomal cosmetic comprising an active polypeptide according to claim 1, characterized in that: the flexible liposome cosmetic containing the active polypeptide is in the dosage form of emulsion, essence, cream, facial mask, freeze-dried powder or facial cleanser.

32. A process for preparing a flexible liposomal cosmetic comprising an active polypeptide according to any one of claims 1-31, characterized in that: the method comprises the following steps:

a. weighing lecithin, sodium deoxycholate or polysorbate 80 and vitamin E, placing into a reaction container, adding a solvent for dissolving, wherein the solvent is a mixture of chloroform and ethanol at a ratio of 1: 1-3 or is ethanol;

b. vacuum rotary evaporating to obtain liposome membrane, and drying;

d. adding the active polypeptide into the blank liposome solution, slightly oscillating, and incubating at room temperature for 30-60 min to obtain the flexible liposome containing the active polypeptide.

33. The method of making a flexible liposome cosmetic product containing an active polypeptide of claim 32, wherein: further comprising e: the liposomes are extruded through a 0.1-0.45 μm polycarbonate membrane 4-8 times.

34. The method of making a flexible liposome cosmetic product containing an active polypeptide of claim 32, wherein: further comprising the step f: according to the dosage form requirement, the flexible liposome is further added with at least one of emulsifying agent, auxiliary emulsifying agent, skin conditioning agent, whitening agent, colorant, humectant, solubilizer, surfactant, antiseptic, aromatic, emollient, anti-acne agent, film forming agent, thickener, pH regulator, buffer, stabilizer or ultraviolet absorbent to prepare cosmetic dosage form.