KR20210040509A - Skin permation enhancing composition of hppln1 using epidermal pentetrating peptide - Google Patents

Abstract

The present invention relates to a transdermal drug delivery composition containing an arginine oligomer that is a transdermal drug delivery peptide skin-penetrating peptide which can excellently increase transdermal absorption by using a cell membrane-penetrating peptide with respect to an HAPLN1 protein which has an excellent skin aging effect but has a molecular weight of 40 KDa and is thus difficult to be absorbed transdermally. More specifically, provided is a protein transdermal drug delivery composition with enhanced transdermal permeability by containing R4 (butyl-D-arginine), R6 (hexa-D-arginine), and R8 (octa-D-arginine) with respect to an HAPLN1 protein.

Description

Skin absorption enhancing composition of HAPLN1 by transdermal penetration peptide {SKIN PERMATION ENHANCING COMPOSITION OF HPPLN1 USING EPIDERMAL PENTETRATING PEPTIDE}

The present invention relates to a composition for enhancing skin absorption of HAPLN1 protein in a transdermal peptide, and more particularly, arginine oligopeptides R4 (butyl-D-arginine), R6 (hexa-D-arginine), R8 (octa-D- arginine) to increase the transdermal permeability of hyaluronic acid and proteoglycan link protein 1 (HAPLN1).

One of the most obvious phenomena due to skin aging is the decrease in the extracellular matrix of the dermal layer, resulting in wrinkles. The extracellular matrix of the dermal layer contains various proteins such as collagen, elastin, and glycoprotein as its main components. More than 90% of these proteins are collagen, of which about 90% is type 1 collagen. It was confirmed that the amount of type 1 collagen in the extracellular matrix of the dermal layer decreased by about 70% due to aging. Hyaluronic acid is a type of polysaccharide composed of amino sugars and uronic acid. It is distributed in all tissues of the living body, and about 50% of the total amount in the body is distributed in the skin. Hyaluronic acid can bind up to 1000 times the mass of water and fills the extracellular matrix of dermal tissue. In addition, hyaluronic acid plays a role in storing or preserving various growth factors by forming an aggregate between various glycoproteins and proteoglycans. According to the results of clinical studies, hyaluronic acid increases the proliferation of dermal fibroblasts, collagen production, and secretion of growth factors, and is known to decrease by more than 40% of the skin with increasing age.

HAPLN1 is a hyaluronic acid and proteoglycan link protein 1 (HAPLN1), which is an effective binding protein for inhibiting skin aging, but has a molecular weight of about 40 kDa and has difficulty in skin absorption. The structure of the skin is divided into epidermis, dermis, and subcutaneous fat, and the epidermis is divided into stratum corneum, transparent layer, granule layer, play layer, and basement layer from the outside. The stratum corneum composed of dead cells is only about 15 μm thick, but 15 to 30 flat and keratinized cell layers are stacked and the density is 1.4 g/cm ² . On the other hand, drug diffusion to the epidermis and dermis under the stratum corneum is relatively easy compared to the stratum corneum, and the blood flow rate of capillaries located in the dermis acts as a sink condition that is sufficiently fast, so passing through the stratum corneum overcomes the transdermal drug delivery system It's the biggest barrier to have.

There are three pathways for transdermal absorption. The first is skin appendages such as pores, the second is transcellual, and the last is skin keratinocytes. It is intercellular through the relatively lipid layer between. Skin appendages occupy 0.1% of the skin's surface area, which contributes to the absorption of drugs very little, and keratinocytes are almost full of keratin, which is a crystal structure, so it is difficult to diffuse and move drugs through the cells. Since the intercellular pathway is used as the main drug absorption pathway and also acts as a barrier to drug permeation, the rate or amount of drug permeation is largely influenced by the distribution coefficient of the drug to intercellular lipids.

Many studies have been conducted for transdermal drug delivery, and the particle size is micronized or nanoparticleized to pass the size of intercellular lipids within 70 nm, liposome structures similar to intercellular lipids, a method using a biotransformation prodrug, and electrical power. A physical method using Tencel, etc. and a method for simultaneous administration of a skin drug metabolism inhibitor, etc., have improved to some extent in transdermal drug delivery, but there is no satisfactory technology yet.

Meanwhile, the stratum corneum of the epidermis located at the outermost part of the skin forms a lamellar structure, which may hinder the penetration of foreign substances during transdermal drug delivery. In order to solve this problem, there is an increasing number of studies on applying Cell Penetrating Peptide to the skin. Peptides act as chemical messengers in the body. Among these, cell-penetrating peptides derived from the protein transduction domain (PTD) are known to be easy to pass through the cell membrane due to the cationic amino acid arginine.

Cell membranes are mainly composed of phospholipids and are limited by substances larger in size than cell membranes. Recently, HIV viruses larger than the cell membrane pores can rapidly penetrate the cell membrane.The reason that the HIV virus is a drug delivery peptide (Trans Activating Transcription, Tat -Peptide) is being made and applied to penetrate the cell membrane. Tat-Peptide is composed of 86 amino acids, and the key amino acid sequence representing drug delivery is [YGRKKRRQRRR], which is amino acids 47 to 57. Arginine (Arg), which has double cationic properties, is known to be the largest core amino acid with six.

As such, it is excellent in inhibiting skin aging, but it contains arginine oligomer, which is a key amino acid sequence of skin-absorbing peptides, targeting HAPLN1 protein, which has a molecular weight of around 40 kDa and is difficult to absorb skin, thereby increasing the skin absorption of HAPLN1 protein. It is believed that the expression of the active ingredient can be maximized.

HAPLN1 protein has an excellent effect in inhibiting skin aging, but it has a disadvantage that it is difficult to absorb skin due to its molecular weight of around 40kDa.

It is an object of the present invention to provide a transdermal drug delivery composition in which the transdermal permeability of HAPLN1 protein is increased by containing arginine oligomer, which is a key amino acid sequence of a skin-absorbing peptide, in order to solve this problem.

In the present invention, it is intended to provide an excellent composition for transdermal drug delivery by increasing the transdermal permeability by containing arginine oligomer, which is a key amino acid sequence of skin-absorbing peptides, relative to HAPLN1 protein with a molecular weight of about 40 kDa.

As a result of the present invention, the transdermal transmittance was relatively increased using Franz Cell Diffusion Methid by containing arginine oligomer, which is a key amino acid sequence of skin-absorbing peptides, targeting HAPLN1 protein.

It relates to a cell membrane-penetrating peptide for increasing the transdermal drug absorption of HAPLN1 protein according to the present invention, and contains arginine oligomers R4, R6, R8, which are key amino acid sequences of the skin-absorbing peptide, to increase the amount of transdermal drug absorption. It is characterized in that it contains 0.00001 to 10.0% by weight of absorbing peptides (R4, R6, R8) at the same time.

In the present invention, a transdermal drug delivery peptide capable of remarkably increasing transdermal absorption by using a cell membrane-penetrating peptide for HAPLN1 protein, which has excellent skin aging effect but has a molecular weight of about 40 KDa, and is difficult to transdermally absorb, and a cosmetic composition. I wanted to complete it.

Example 1: Arginine Oligomer (R4, R6, R8)

The reason that the HIV virus larger than the cell membrane pore can rapidly penetrate the cell membrane is that the HIV virus creates a drug delivery peptide (Trans Activating Transcription, Tat-Peptide) to penetrate the cell membrane, and Tat-Peptide is 86 amino acids. It is composed of and the key amino acid sequence representing drug delivery is [YGRKKRRQRRR, which is amino acids 47 to 57. Arginine (Arg), which has double cationic properties, is known to be the largest core amino acid with six. Arginine with double cationic properties (Arg, R) was used as a skin-absorbing peptide using R4 [RRRR] containing 4, R6 containing 6 [RRRRRR], and R8 containing 8 [RRRRRRRR]. .

Example 2. The HAPLN1 protein was subjected to an experiment for measuring skin absorption by targeting functional proteins known to have an effect on improving wrinkles, etc., respectively.

Example 3. Skin absorption measurement test (Franz Cell)

For skin penetration experiments, fill the receptor chamber with 5 mL of the receptor phase (HCO-60: ethanol: PBS = 2: 20: 78 (w/w/w %)), and then the stratum corneum of the prepared pig skin (Ginia Pig, thickness 300 μm). It was fixed between the donor and receptor phases of a Franz diffusion cell (Standard diameter 9mm, Receiver 5ml, Permegear) facing upward. At this time, the temperature was maintained at 37° C. using a constant temperature water bath, and the mixture was stirred at 150 rpm for 24 h. Prepare an aqueous solution containing 50 ppm (0.005%) of each HAPLN1 protein and 50 ppm (0.005%) of Arginine Oligomer, R4, R6, and R8, respectively, and 0.2 mL of this solution was added dropwise to the donor, followed by 24 For h, 0.5 mL of the receptor phase was taken through the sampling port. Immediately after collection, the same amount of receptor solution was added to the receptor chamber. The collected receptor solution was analyzed at each maximum absorption wavelength by HPLC as the content of each peptide (transdermal) that passed through the skin.

The skin absorption amount of HAPLN1 protein was greatly increased depending on the presence or absence of Arginine Oligomer, R4, R6, R8, and the results are presented in Table 1, and the present invention was completed.

Claims (1)

Contains 0.0001 to 10.0% of the total weight of the HAPLN1 protein and 0.0001 to 10.0% by weight of skin-penetrating peptides R4 (butyl-D-arginine), R6 (hexa-D-arginine), and R8 (octa-D-arginine). Transdermal drug delivery cosmetic composition.