KR101837023B1 - Cosmetic nanoliposome composition containg peptides for inhibiting degradation of collagen and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a nano-sized liposome cosmetic composition containing a collagen degradation inhibiting peptide. The liposome cosmetic composition of the present invention comprises a peptide having low turbidity, a collagen degradation inhibiting peptide as a stable nano-sized liposome, .
The cosmetic composition of the present invention contains an effective ingredient in a nano-sized liposome of stable size, which is excellent in skin beauty, low cytotoxicity and storage stability.
The present invention also provides a method for producing the above-described nano-sized liposome cosmetic composition containing an active ingredient in a high content.

Description

TECHNICAL FIELD The present invention relates to a nano-sized liposome cosmetic composition containing a collagen degradation inhibiting peptide and a method for producing the same, and a method for producing the same. BACKGROUND ART < RTI ID = 0.0 >

The present invention relates to a nano-sized liposome cosmetic composition containing a collagen degradation inhibiting peptide and a method for producing the same, more particularly, to a nano-sized liposome cosmetic composition which is a peptide composition stabilized by encapsulating a specific functional peptide into a nanoliposome And a method for producing the same.

Skin is an important organ that plays various roles such as protective function, barrier function, temperature control function, excretion function, and respiratory function, which are composed of epidermis, dermis, and subcutaneous tissue. However, due to the various external environments, the thickness of epidermis, dermis, and subcutaneous tissue becomes thinner, and as the composition and content of the lipid barrier, which functions as a skin barrier, changes, its function rapidly decreases and aging occurs. Molecular biologically, skin aging is known to be a major cause of collagen reduction in the skin dermis.

Collagen is 70 ~ 80% of the skin's dermis, and it is a typical substance that is involved in maintaining the elasticity of the skin. According to the research results of the Korean Society of Cosmetic Scientists Volume 38, Issue 4, Vol. 87, it is known that collagen peptides have skin recovery and skin regeneration effects.

As collagen ages, its production is decreased, and degradation is promoted by an external environment such as ultraviolet rays, which causes wrinkles of the skin, and aging of the skin is promoted.

In order to solve such problems as skin aging and wrinkle improvement, various studies on the change of collagen have been carried out. Especially, it has been reported that an enzyme called MMP (Matrix metalloproteinase) decomposes collagen, -0514315, there have been various studies on a substance that inhibits the expression of MMP enzyme.

In recent years, studies have been conducted to stabilize active ingredients. Such stabilization methods have been used for about 10 years, not only in the cosmetics industry, but also in pharmaceuticals, electronics, communications, Research on nanotechnology has been actively pursued in all industrial fields.

Nanotechnology is a technology that efficiently transports the drug concentration to a nano-sized capsule efficiently enough to exhibit its efficacy. It is largely divided into an emulsion and a liposome. The emulsion encapsulates a lipophilic substance, and the liposome encapsulates a hydrophilic substance into a skin This technology can be applied according to each characteristic.

In such a technique, it is required to stabilize the active ingredient at a high concentration and a high content. The present inventors have developed the collagen degradation inhibitory peptide as an active ingredient and stably formulate it into a nano-sized liposome capsule at a high content to complete the present invention Respectively.

It is an object of the present invention to provide a nano-sized liposome cosmetic composition containing a collagen degradation inhibiting peptide.

More specifically, the collagen decomposition inhibiting peptide comprises a peptide of the SRPQGPFL sequence and is excellent in stability and storage activity.

Another object of the present invention is to provide a method for producing a nano-sized liposome cosmetic composition which is obtained by stabilizing a collagen decomposition inhibiting peptide by a liposome method.

In order to achieve the object of the present invention, there is provided a nano-sized liposome cosmetic composition comprising a collagen degradation-inhibiting peptide, wherein the liposome cosmetic composition comprises a lipid stabilizer and a lipid thickener.

The collagen decomposition inhibiting peptide is characterized by containing an effective physiologically active substance and an MMP activity inhibiting peptide.

A GG linker may be introduced between the effective physiologically active substance and the MMP activity inhibitory peptide, and the effective physiologically active substance is one effective physiologically active substance selected from the group consisting of lactobionic acid and biotin, and the MMP activity inhibitory peptide is SRPQGPFL And VHTVCDPLFEAVGKI. ≪ / RTI >

The collagen degradation inhibiting peptide is preferably lactobionoyl-GG-SRPQGPFL.

The collagen degradation inhibiting peptide is contained in an amount of 1.0 to 10.0% by weight as a 0.5% solution, and the lipid stabilizing part includes purified water, adenosine, and kavasome. The lipid thickening part may include 1,3-butylene glycol, phospholipids, ceramide, cholesterol, , And useful antioxidants.

The nano-sized liposome has a particle size of 100 to 300 nm.

In order to achieve another object of the present invention, there is provided a method for preparing a lipid-stabilizing agent, comprising the steps of: (1) (2) preparing a lipid thickener for dissolving phospholipids, ceramide, cholesterol, stearic acid and an antioxidant in a solvent having a hydroxyl group by heating at 75 캜; (3) a first concentrated phase forming step in which the dissolved lipid thickening part is put into the dissolved lipid stabilizing part and stirred at 2,000 rpm to make it uniform; (4) The first concentrated phase formed was added to a commercially available high-pressure emulsifier after adding collagen decomposition-inhibiting peptide at a temperature of 45 ° C, and treated three times at a pressure of 1,000 bar to prepare a nano-concentrate composition having a nano- A second concentrated phase forming step of obtaining a second concentrated phase; And (5) a second concentrated phase stabilizing step of stabilizing the second concentrated phase by cooling to 25 to 40 캜. The present invention also provides a method for producing a nano-sized liposome cosmetic composition.

The present invention relates to a nano-sized liposome cosmetic composition which contains a high content of collagen degradation-inhibiting peptide which is excellent in skin cosmetics, has no cytotoxicity, has good solubility in water, excellent storage stability and storage activity, Stabilized.

In the present invention, the peptide is in the form of a nano-sized liposome and has excellent skin absorption.

1 shows the structure of Lactobionoyl-SRPQGPFL of the present invention,
Figure 2 shows the structure of Biotinoyl-SRPQGPFL of the present invention,
FIG. 3 shows the structure of Lactobionoyl-GG-SRPQGPFL of the present invention,
Figure 4 shows the structure of the biotinoyl-GG-SRPQGPFL of the present invention,
FIG. 5 shows the structure of Lactobionoyl-GG-VHTVCDPLFEAVGKI of the present invention,
FIG. 6 shows the results of suppressing MMP-1 activity of the collagen degradation inhibiting peptide of the present invention,
7 is a cytotoxicity analysis result of the collagen degradation inhibiting peptide of the present invention,
8 is a result of particle size analysis of the nano-sized liposome cosmetic composition of the present invention.

The present invention relates to a nano-sized liposome cosmetic composition containing a collagen decomposition inhibiting peptide, and includes a lipid stabilizing portion and a lipid thickening portion. Hereinafter, the present invention will be described in more detail.

1. Nano-sized liposome cosmetic composition

The nano-sized liposome cosmetic composition of the present invention is composed of a lipid stabilizing part, a lipid thickening part and a peptide.

First, the lipid stabilizing part contains 2.0 wt% of adenosine and 0.5 wt% of kavasome in the purified water. The adenosine can be used without limitation as long as it is adenosine product of ZMC of Japan and commercially available, and kavasome is an extract of leaves, roots, and stem of Kaba, which can be used as a kavasome concentrate product of Korea Chemtrot Co., The lipid stabilizing part includes a hydrophilic active material.

The lipid concentrate comprises phospholipids, ceramides, cholesterol, fatty acids and an antioxidant, more preferably 20 wt% 1,3-butylene glycol, 3.0 wt% phospholipid, 1.0 wt% ceramide, 1.0 wt% Cholesterol, 0.5% by weight stearic acid, 0.05% by weight of an antioxidant.

The collagen degradation inhibiting peptide is selected from peptide candidates inhibiting MMP activity, and may be a VHTVCDPLFEAVGKI peptide or a SRPQGPFL peptide.

In particular, the SRPQGPFL peptide is a peptide consistent with the CD44 binding peptide sequence derived from PEX (hemoPEXin domain 9), and is reported to have an EC50 of 11 μM.

The collagen decomposition inhibiting peptide may further comprise an effective physiologically active substance in the MMP activity inhibiting peptide. Effective biologically active substances can use biotin, which has been shown to be effective in promoting collagen biosynthesis and lactobionic acid with high solubility (50 mg / ml in DW), as well as inhibiting MMP activity.

In addition, a linker may be introduced between the MMP activity inhibiting peptide and the active physiologically active substance in order to enhance the activity of the collagen degradation inhibiting peptide. The linker may be selected from GG, AA, KK, and PEG, and the linker may be selected from among GG, AA, KK, and PEG.

Lactobionoyl-SRPQGPFL, Biotinoyl-SRPQGPFL, Lactobionoyl-GG-SRPQGPFL, Biotinoyl-GG-SRPQGPFL and Lactobionoyl-GG-VHTVCDPLFEAVGKI as the collagen degradation inhibiting peptide are shown in FIG. 1 to FIG. 5, respectively.

The collagen degradation-inhibiting peptide was cultured for 96 hours in human dermal fibroblast cells cultured on a 96-well plate. After culturing for 96 hours, trypsinization of the medium was performed to activate MMP-1 in the culture medium and reacted with thiopeptolide As shown in FIG. 6, the lactobionic acid-GGSRPQGPFL peptide showed a 50% inhibitory effect on MMP activity at 100 .mu.M, which is preferable.

The collagen degradation inhibiting peptide can be synthesized by sequentially polymerizing the peptides using the n-term terminal of each peptide based on the peptide monomer introduced into the resin. In this process, the Fmoc elimination process and the coupling process are repeated Can be synthesized by proceeding.

The collagen degradation inhibitor peptide was cultured on a 96-well plate containing human dermal fibroblast cells and the collagen degradation inhibitor peptide derivative was treated for 48 hours. Cell viability was assayed using the MIT assay. As a result, peptide derivatives with Lactobionic acid and biotin Are excellent because they do not show cytotoxicity up to 100 [mu] M (Fig. 7).

The collagen degradation inhibiting peptide may be prepared in a 0.5% solution and may be contained in an amount of 1.0 to 10.0% by weight.

2. Manufacturing method of cosmetic composition

The present invention relates to a method for producing a cosmetic composition which stabilizes collagen degradation inhibition peptide with nano-sized liposomes. In particular, the present invention relates to a method for stabilizing a collagen degradation inhibitor peptide with a nanoliposome by incorporating the collagen degradation inhibitor peptide in the highest amount.

(1) a lipid stabilizing portion preparation step in which the lipid stabilizing portion is dissolved by heating to 75 캜; (2) preparing a lipid thickener for dissolving phospholipids, ceramide, cholesterol, stearic acid and an antioxidant in a solvent having a hydroxyl group by heating at 75 캜; (3) a first concentrated phase forming step in which the dissolved lipid thickening part is put into the dissolved lipid stabilizing part and stirred at 2,000 rpm to make it uniform; (4) adding the peptide at a temperature of 45 ° C to the first concentrated phase, adding the peptide to the commercialized high pressure emulsifier, and treating the mixture at a pressure of 1,000 bar three times to obtain a nano-thickened composition having a nano-scale of 100 to 300 nm A second concentrated phase forming step; (5) a second concentrated phase stabilizing step of stabilizing the second concentrated phase by cooling to 25 to 40 캜; and (5) a second concentrated phase stabilizing step of cooling the second concentrated phase to 25 to 40 캜.

(1) The step of preparing the dissolved lipid stabilizing part by heating at 75 DEG C is a step of dissolving adenosine and kavasome in warmed purified water, preferably adding adenosine 2.0 weight% and kavasome 0.5 wt% .

(2) The lipid thickener preparation step for dissolving phospholipids, ceramide, cholesterol, stearic acid and an antioxidant in a solvent having a hydroxyl group by heating at 75 DEG C is preferably a step of adding 20 wt% of 1,3-butylene glycol to 3.0 wt% , 1.0% by weight of ceramide, 1.0% by weight of cholesterol, 0.5% by weight of stearic acid and 0.05% by weight of an antioxidant.

(3) The dissolved lipid thickening part may be added to the dissolved lipid stabilizing part and stirred at 2,000 rpm to make the first concentrated phase forming step, a conventional agitator may be used.

(4) The first concentrated phase formed was added to a commercially available high-pressure emulsifier after adding collagen decomposition-inhibiting peptide at a temperature of 45 ° C, and treated three times at a pressure of 1,000 bar to prepare a nano-concentrate composition having a nano- The collagen decomposition inhibiting peptide is added in an amount of 1.0 to 10.0% by weight and the step of forming the concentrated base with a size of 2 to 5 탆 by using an in-line mixer before cooling .

(5) The second concentrated phase stabilization step in which the second concentrated phase is stabilized by cooling to 25 to 40 DEG C is such that the collagen decomposition inhibiting peptide is uniformly encapsulated in the phospholipid liposome by cooling the concentrated phase slowly, It can not be uniformly encapsulated. If the temperature exceeds 40 ° C, the structure of the phospholipid liposome is modified to fail to form a nano-sized homogeneous liposome.

The method for producing the nano-sized liposome cosmetic composition of the present invention may be applied in different production conditions so as to be mass-produced in an actual production process. Specifically, phospholipid, ceramide, and cholesterol may be added to a solvent containing water in a separate dissolution tank. Lt; 0 > C to make a lipid thickening part, and completely dissolving the lipid-stabilized part at 75 DEG C in a separate melting tank; Adding a lipid thickening part to the lipid stabilizing part through an emulsifier and conducting primary emulsification to make a bulky primary concentrated base; Making a second enrichment base having a size of 2 to 5 mu m using an inline mixer; After cooling to 45 to 50 캜, collagen degradation inhibiting peptide was added and the mixture was added to a high-pressure emulsifier (Microfluidics, Model: M-110F) and treated at a pressure of 9,000 bar twice. And the like.

The validation for mass production is to increase lactobionoyl-GGSRPQGPFL component through the manufacturing and aging process of nanocapsules using the optimal micro-scale concentration base and microfluidizer while increasing the production volume from 1L scale to 5L, 10L, 30L scale sequentially The optimal process conditions for the introduction of lactobionoyl-GGSRPQGPFL into the nanocapsule structure with stable and uniform size are established.

The content of the ingredients for preparing the concentrated nano-sized liposome cosmetic composition containing lactobionoyl-GGSRPQGPFL is shown in Table 1.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are intended to further illustrate the present invention, and the scope of the present invention is not limited to these examples.

≪ Examples 1 to 4 > Preparation of nano-sized liposome cosmetic composition

The lipid-stabilized portion dissolved in the composition as shown in [Table 2] was prepared by dissolving phospholipids, ceramide, cholesterol, stearic acid and an oil-soluble antioxidant by heating at 75 ° C in 1,3-butylene glycol, Prepare to bear. The dissolved lipid-enriched portion was added to the dissolved lipid-stabilizing portion and stirred at 2,000 rpm to homogenize to form a first concentrated phase. Then, 0.5% solubilized lactobionoyl-GGSRPQGPFL at 45 ° C was added to the content of [Table 2] Pressure emulsifier and treated at a pressure of 1,000 bar three times to obtain a nano-thickened composition having a nano-scale of 100 to 300 nm. The nano-thickened composition was cooled to 25 to 40 캜 and stabilized to prepare a nano-sized liposome cosmetic composition.

<Comparative Examples 1 and 2> Preparation of nano-sized liposome cosmetic composition

Was performed in the same manner as in Example 1 except that the composition and the content thereof shown in Table 2 below were used to prepare a nano-sized liposome cosmetic composition containing lactobionoyl-GGSRPQGPFL 0.5% sol.

division Component (% by weight) Example 1 Example 2 Example 3 Example 4 Comparative Example 1 Comparative Example 2 Lipid
Stabilizing portion Purified water to.100 to.100 to.100 to.100 to.100 to.100 Adenosine 2.0 2.0 2.0 2.0 2.0 2.0 kavasome 0.5 0.5 0.5 0.5 0.5 0.5 Peptides lactobionoyl-GG-SRPQGPFL 0.5% sol. 1.0 3.0 5.0 10.0 20.0 30.0 Lipid
Concentrated portion 1,3-butylene
Glycol 20.0 20.0 20.0 20.0 20.0 20.0 Phospholipids 3.0 3.0 3.0 3.0 3.0 3.0 Ceramide 1.0 1.0 1.0 1.0 1.0 1.0 cholesterol 1.0 1.0 1.0 1.0 1.0 1.0 Stearic acid 0.5 0.5 0.5 0.5 0.5 0.5 Availability antioxidant 0.05 0.05 0.05 0.05 0.05 0.05 1) 1,3-butylene glycol: DAICEL Co., Ltd., Japan
2) Phospholipids: Lipoid S75-3, Germany, Lipoid
3) Ceramide: DS-CERAMIDE, Korea, Doosan Biotech
4) Cholesterol: Cholesterol NF, United States of America, Solvay (Solvay)
5) Stearic acid: Emersol 132 (stearic acid: palmitic acid = 46: 54), Germany, Cognis
6) Availability Antioxidant: Dibutylhydroxytoluene, Ueno, Japan, Inc.
7) Water-soluble antioxidants: sodium pyrosulfite,
8) Acetyltetrapeptide: BCM-AWA, Spain, LIPOTEC
9) Kavasome: Kavasome Concentrate,

&Lt; Experimental Example 1 >

The results of observing the acidity and transparency of the nano-sized liposome cosmetic composition prepared in Examples 1 to 4 and Comparative Examples 1 and 2 are shown in [Table 3].

Properties
pH 6.24 6.24 6.59 6.12 6.31 6.42 transparency ◎ ◎ ◎ ◎ ○ △ Transparency:? (Transparent),? (Translucent),? (Translucent-turbid), x (milky white)

All of the cosmetic compositions prepared in the above Table 3 were found to be suitable for application to skin with an acidity of pH 6.0 to 7.0, but lactobionoyl-GG-SRPQGPFL 0.5% sol. It is confirmed that the compositions of Comparative Examples 1 and 2 having a content of more than 10.0% become cloudy.

<Experimental Example 2> Particle size analysis and stability test

Particle size analyzer using light scattering method was used to check the size and particle size distribution of nano-sized multi-layered liposome which is not confirmed by general optical microscope. Using Helos model equipment of Sympatec, Germany. Stability tests were performed at 5 ℃, 25 ℃ and 45 ℃ for 1 month.

The results of the visual observation in Table 4 were analyzed and verified using a particle size analyzer using a light scattering method. The results of the particle size analysis are shown in FIG. 8.

Measurement sample Example 1 Example 2 Example 3 Example 4 Comparative Example 1 Comparative Example 2 Particle size
(Early) 98nm 198nm 196 nm 190 nm 608 nm 1256 nm transparency
(Early) ◎ ◎ ◎ ◎ ○ ○ Particle size (45 ℃, 1 month) 107 nm 260nm 201nm 256nm 542 nm Two week separation transparency
(45 占 폚, 1 month) ◎ ◎ ◎ ◎ ○ X Particle Cake
(5 占 폚, 1 month) 80nm 180nm 165 nm 215 nm 896 nm 896 nm transparency
(5 占 폚, 1 month) ◎ ◎ ◎ ◎ ○ ○ Particle Cake
(Cyc, 1 month) 89 nm 197nm 204 nm 203 nm 966 nm Two week separation transparency
(Cyc, 1 month) ◎ ◎ ◎ ◎ ○ X Particle Cake
(25 占 폚, 1 month) 197nm 160 nm 182 nm 191 nm 756 nm 966 nm transparency
(25 占 폚, 1 month) ◎ ◎ ◎ ◎ ○ ○ Transparency:? (Transparent),? (Translucent),? (Translucent-turbid), x (milky white)

When the content of lactobionoyl-GG-SRPQGPFL 0.5% sol is more than 10.0%, it can be seen that in Example 4, 10.0% of lactobionoyl-GG-SRPQGPFL 0.5% sol. It is possible to confirm that the encapsulation of the resin is not stable and is separated and that the properties are cloudy.

Claims (9)

A nano-sized liposome containing 1.0% by weight to 10.0% by weight of a 0.5% by weight solution of a collagen decomposition-inhibiting peptide (lactobionoyl-GG-SRPQGPFL) A cosmetic composition comprising:
Wherein the liposome comprises a lipid stabilizing portion and a lipid thickening portion,
Wherein the lipid-stabilizing moiety comprises purified water and adenosine,
Characterized in that the lipid concentrate comprises 1,3-butylene glycol, phospholipids, ceramides, cholesterol, stearic acid and an antioxidant.
A cosmetic composition comprising a nano-sized liposome.

[Chemical Formula 1]

delete delete delete delete delete delete The cosmetic composition according to claim 1, wherein the nano-sized liposome has a particle size of 100 to 300 nm. delete

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