JP2016079182A - Drug delivery carrier in which cell-penetrating peptide comprising macromolecule is introduced - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a drug delivery carrier in which a cell-penetrating peptide comprising hydrophilic macromolecules is introduced.SOLUTION: The invention provides a composition comprising: a drug delivery carrier containing a lipid structure or polymer particles, which are covalently bonded to AP-GRR peptide that is a cell penetration peptide or the peptide chain containing the peptide or deformed by the peptide chain comprising the peptide; and a bioactive component collected into the carrier thereof, wherein by effectively delivering, to in vivo cells, macromolecules which are difficult to introduce into cells, the peptide can enhance the bioavailability of the macromolecules, the cell penetration peptide has a sequence of Gly (Arg) nGly Tyr Lys Cys (n=1-20), and the number average molecular weight or weight average molecular weight of the water-soluble or water-insoluble macromolecule is 5,000 Da or more.SELECTED DRAWING: None

Description

  The present specification relates to a drug delivery carrier into which a cell-penetrating peptide containing a macromolecule is introduced.

  In general, hydrophilic and huge substances cannot enter cells by a barrier called a cell membrane. The cell membrane blocks macromolecules such as peptides, proteins, and nucleic acids from entering the cell, and even if it enters the cell by a physiological mechanism called endocytosis by the cell membrane receptor, the cell lysosome ( Because it is fused with lysosomal compartments and eventually decomposes, there are many restrictions in the treatment and prevention of diseases using these.

  Therefore, development of a novel preparation capable of effectively delivering a biological substance into a living body and having no cytotoxicity is required and urgently required. In recent years, several new alternatives have been reported, of which cell permeable peptide has previously been difficult to use as a drug due to low cell membrane permeability and fast in vivo half-life. It can increase the utility value of macromolecules such as protein and gene, and has attracted much attention.

  Peptides having such a cell membrane permeability function are membrane-permeable peptides mainly derived from proteins, and can be roughly classified into three types. The first is a homeodomain-derived peptide penetratin, which is SEQ ID NO: 2 (Drosophila melanogaster, amino acid sequence: Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys ) Amino acid sequence. It was discovered from the homeodomain of Antennapedia of Drosophila homeoprotein (A. Joliot et al., Proc. Natl. Acad. Sci. USA, (1991) 88, 1864. ). Here, the homeoprotein is a kind of transcription factor and has a structure of about 60 amino acids that can bind to DNA called homeodomain. The second is 49- of Tat protein, which is a transcription-related protein of human immunodeficiency virus (HIV-1), which is a strain that causes acquired immune deficiency syndrome (AIDS). A Tat49-57 peptide existing between No. 57, which has the amino acid sequence of SEQ ID NO: 3 (Human immunodeficiency virus type 1, amino acid sequence: Arg Lys Lys Arg Arg Gln Arg Arg Arg) (PA Wender et al PNAS (2000) 97, 24, 13003-13008). The third is a peptide based on a membrane translocating sequence (hereinafter referred to as MTS) or a signal sequence, in which a newly synthesized protein by RNA is adapted to a suitable organelle in vivo. MTS, which is recognized by an acceptor protein that helps to be located in the membrane of (organelle) and combined with a nuclear localization signal (hereinafter referred to as NLS), is a cell of several types It has become clear that it accumulates in the cell nucleus through the cell membrane. For example, nuclear transcription factor kappa B (NF-κB), monkey virus 40T antigen (Simian virus 40 (SV40) T-antige) or Kaposi sarcoma fibroblast growth factor 1 In the MTS derived from a hydrophobic region of a signal sequence such as K-FGF, human β3 integrin, HIV-1 gp41, etc., bound to an NLS peptide derived from factor 1 (hereinafter referred to as K-FGF) (Y. Lin et al., J. Biol. Chem. (1996) 271, 5305; X. Lin et al., Proc. Natl. Acad. Sci. USA (1996) 93 MC Morris et al. Nucleic Acids Res. (1997) 25, 2730; L. Zhang et al. Proc. Natl. Acad. Sci. USA (1998) 95, 9184; Chaloin et al., Biochiem. Res. Commun. (1998) 243, 601; Y. Lin et al., J. Biol. Chem. (1995) 270, 14255).

A. Joliot et al., Proc. Natl. Acad. Sci. U.S.A., (1991) 88, 1864 P. A. Wender et al., PNAS (2000) 97, 24, 13003-13008 Y. Lin et al., J. Biol. Chem. (1996) 271, 5305 X. Lin et al., Proc. Natl. Acad. Sci. U. S. A. (1996) 93, 11819 M. C. Morris et al. Nucleic Acids Res. (1997) 25, 2730 L. Zhang et al. Proc. Natl. Acad. Sci. U. S. A. (1998) 95, 9184 Chaloin et al., Biochiem. Biochim. Res. Commun. (1998) 243, 601 Y. Lin et al., J. Biol. Chem. (1995) 270, 14255

  An object of the present invention is to provide a composition comprising a cell-penetrating peptide and a physiologically active ingredient in order to deliver a macromolecular substance that is difficult to be taken into a cell.

  In order to achieve the above object, in one aspect, the present invention provides a drug delivery carrier comprising a lipid structure or a polymer particle covalently bound to an AP-GRR peptide or a peptide chain comprising the peptide, and a trap within the carrier. A composition comprising the collected bioactive ingredients is provided.

  According to the drug delivery carrier according to one aspect of the present invention, a macromolecular substance having a large molecular weight can be delivered into a cell of a physiologically active ingredient to be delivered by introducing an AP-GRR peptide that effectively exhibits a membrane permeability function. There is an effect that the amount delivered to can be greatly increased. Therefore, the drug delivery carrier according to one aspect of the present invention has overcome the shortcoming that a giant physiologically active ingredient such as a polysaccharide, an enzyme, a peptide, a drug, or a protein is difficult to be delivered into a cell.

It is the photograph which observed the liposome aqueous solution containing the comparative example of the drug delivery carrier based on this invention with the transmission electron microscope. It is the photograph which observed the drug delivery carrier based on this invention with the transmission electron microscope. It is a graph which shows the result of having processed the drug delivery support | carrier which carry | supported the fluorescent substance which concerns on 1 side of this invention to a cell, and having analyzed the quantity of the fluorescent substance transferred into the cell by FACS. It is a graph which shows the result of having processed the drug delivery support | carrier which carry | supported the fluorescent substance which concerns on 1 side of this invention to a cell, and having analyzed the quantity of the fluorescent substance transferred into the cell by FACS. It is a table | surface which shows the result of having processed the drug delivery support | carrier which carry | supported the fluorescent substance which concerns on 1 side of this invention to a cell, and analyzed the quantity of the fluorescent substance transferred into the cell by FACS. It is a photograph which shows the image result which processed the drug delivery support | carrier which carry | supported the rhodamine B which concerns on 1 side of this invention to a cell, and observed this with the confocal laser scanning microscope (confocal laser scanning microscope, CLSM). It is a photograph which shows the image result which processed the drug delivery support | carrier which carry | supported the dextran-RITC which concerns on one side of this invention to a cell, and observed this with the confocal laser scanning microscope (confocal laser scanning microscope, CLSM). The microscopic image result which shows the skin absorption experiment result of the sample which applied the rhodamine B and the dextran-RITC to PBS and the general lipozone, respectively is shown. The microscopic image result which shows the skin absorption experiment result of the sample which applied the rhodamine B and the dextran-RITC to the drug delivery carrier which concerns on PBS and 1 side of this invention, respectively is shown. It is the graph which showed the quantitative result of the skin absorption experiment with respect to rhodamine B with respect to each sample. It is the graph which showed the quantitative result of the skin absorption experiment with respect to rhodamine B with respect to each sample. It is the graph which showed the quantitative result of the skin absorption experiment with respect to dextran-RITC with respect to each sample. It is the graph which showed the quantitative result of the skin absorption experiment with respect to dextran-RITC with respect to each sample.

  In one aspect, the present invention may relate to a drug delivery carrier comprising a lipid structure or a polymer particle covalently bound to a cell-penetrating peptide or a peptide chain containing the same.

  In one aspect of the present invention, the lipid structure or polymer particle may have a structure in which a physiologically active component is collected inside the structure.

  In one aspect of the invention, the lipid structure may be a lipid construct.

  In one aspect of the present invention, the polymer particles may be a polymer structure or a polymer construct.

  In one aspect of the present invention, the physiologically active ingredient may be a macromolecule having a number average molecular weight or a weight average molecular weight of 500 Da or more. Specifically, the physiologically active ingredient may be a water-insoluble or water-soluble macromolecule. .

  In one aspect of the invention, when the drug delivery carrier comprises a lipid structure, the bioactive component may be a water soluble macromolecule.

  In one aspect of the present invention, when the drug delivery carrier includes polymer particles, the bioactive component may be a water-insoluble macromolecule.

  In one aspect of the present invention, the cell penetrating peptide may be an AP-GRR peptide having a sequence of Gly (Arg) n Gly Tyr Lys Cys (n = 1 to 20).

  In one aspect of the present invention, in the cell-penetrating peptide, n may be 3 to 9.

  In one aspect of the present invention, the cell-penetrating peptide may comprise the sequence of SEQ ID NO: 1 (amino acid sequence: Gly Arg Arg Arg Arg Arg Arg Arg Arg Gly Tyr Lys Cys).

  Specifically, in one aspect of the present invention, the cell penetrating peptide may be the sequence of SEQ ID NO: 1 (Gly Arg Arg Arg Arg Arg Arg Arg Arg Gly Tyr Lys Cys).

  In one aspect of the present invention, the lipid structure or polymer particle may contain an amphiphilic polymer.

  In one aspect of the present invention, when the polymer particle further includes an amphiphilic polymer, the polymer particle may be an amphiphilic polymer or produced therefrom.

  In one aspect of the present invention, the cell-penetrating peptide or peptide chain containing the same may be covalently bound to an amphiphilic polymer.

  In one aspect of the present invention, a covalent bond between a cell-penetrating peptide or a peptide chain containing the peptide and a lipid structure or polymer particle is a covalent bond generated by a bond between a maleimide group and a thiol group. It may be. Such covalent bonds are fairly stable and are well known to ordinary engineers.

  In one aspect of the present invention, when a cell-penetrating peptide or a peptide chain containing the cell-penetrating peptide is bonded to a lipid structure, a polymer particle, or an amphiphilic polymer contained therein, a bond between a maleimide group and a thiol group However, it is not necessarily limited to this as long as it is a stable covalent bond. Since the cell-penetrating peptide according to one aspect of the present invention has a thiol group, the cell-penetrating peptide can be obtained by introducing a maleimide group into a lipid structure, a polymer particle, or an amphiphilic polymer contained therein. Can be covalently linked. The introduction of the maleimide group into the lipid structure, polymer particle, or the amphiphilic polymer contained in the lipid structure according to one aspect of the present invention involves bonding the maleimide group to a carboxyl group of the lipid structure or the like to be introduced. Can be done by.

  In one aspect of the present invention, the amphiphilic polymer includes an alkylated hyaluronic acid having an alkyl group attached to a hyaluronic acid side chain, poly (methacrylic acid-co-n-alkyl) represented by Formula 1. It may be at least one selected from the group consisting of a methacrylate) random copolymer and a poly (hydroxyethyl methacrylate-co-n-alkyl methacrylate) random copolymer represented by Chemical Formula 2, but is not necessarily limited thereto. Is not to be done. In one aspect of the present invention, the amphiphilic polymer may be a general acrylate polymer produced by polymerization using a general free radical thermal initiation method.

[In the above formulas 1 and 2, n = 7-22,
x: y is 90: 10-50: 50 by molar ratio. ]

  In one aspect of the present invention, the poly (methacrylic acid-co-n-alkyl methacrylate) random copolymer may be composed of two types of monomers, methacrylic acid and n-alkyl methacrylate. Such a polymer is produced by polymerization by a general free radical thermal initiation method, and an anionic or cationic polymerization may be used to adjust the molecular weight distribution.

  In one aspect of the present invention, the poly (hydroxyethyl methacrylate-co-n-alkyl methacrylate) random copolymer may be composed of two types of monomers, hydroxyethyl methacrylate and n-alkyl methacrylate. Such a polymer is produced by polymerization by a general free radical thermal initiation method, and an anionic or cationic polymerization may be used to adjust the molecular weight distribution.

  In an aspect of the present invention, in Chemical Formulas 1 and 2, n may be an integer of 5 or more and 30 or less, specifically an integer between 7 and 22, more specifically 11-22. It may be an integer between.

  In one aspect of the present invention, in Formulas 1 and 2, x: y may be an integer between 50 and 90 in molar ratio: an integer between 10 and 50, specifically 90:10, 85: It may be 15, 70:30, 60:40, or 50:50, preferably 85:15 to 70:30.

  In one aspect of the present invention, the molecular weight of the amphiphilic polymer having a structure represented by Chemical Formula 1 or 2 affects the structure of the polymer-liposome nanocomposite, and the drug delivery according to one aspect of the present invention. The molecular weight of the polymer used for the carrier is 5,000 to 100,000, preferably 10,000 to 50,000, based on the number average molecular weight.

  In the drug delivery carrier according to one aspect of the present invention, the lipid structure can have a more stable structure due to the amphiphilic polymer.

  Liposomes based on lipid-cholesterol need to ensure stability in the dosage form in order to be used in cosmetics and external preparations for skin, but are easily controlled by various surfactants in the dosage form. There is a problem of eliminating the structure. By introducing a polymer having amphiphilic properties into such liposomes, structurally unstable disadvantages can be overcome to some extent. The polymer-liposome complex into which the amphiphilic polymer is introduced retains a similar form to the liposome based on lipid-cholesterol, and the hydrophobic part of the amphiphilic polymer contains lipid and lipid-cholesterol. It will have a structure associated together between the lipid bilayers as the basis. As a result, the lipid bilayer is tightly connected to protect the outer wall, and the role of maintaining the liposome structure stably from elements such as salts or surfactants that destabilize the various liposome structures present in the aqueous phase. Will do.

  In the drug delivery carrier according to one aspect of the present invention, the molecular weight range of the polymer that may be used as the amphiphilic polymer is 5,000 Da to 200,000 Da based on the number average molecular weight. Preferably, it may be 10,000 to 100,000 Da. The molecular weight of the amphiphilic polymer is 1,000 Da or more, 2,000 Da or more, 3,000 Da or more, 4,000 Da or more, 5,000 Da or more, 6,000 Da or more, 7,000 Da or more, 8,000 Da or more, 9,000 Da or more, 10,000 Da or more, 11,000 Da or more, 12,000 Da or more, 13,000 Da or more, 14,000 Da or more, 15,000 Da or more, 20,000 Da or more, 30,000 Da or more, 50,000 Da or more, Alternatively, it is 100,000 Da or more, or 200,000 Da or less, 150,000 Da or less, 100,000 Da or less, 90,000 Da or less, 80,000 Da or less, 70,000 Da or less, 60,000 Da or less, 50,000 Da or less, 40 30,000 Da or less, 30,000 Da Lower, 20,000 Da or less, 10,000 Da or less, 5,000 Da or less, 3,000 Da or less, or 1,000Da may be less, but is not necessarily limited thereto.

  In one aspect of the present invention, the hydrophobic portion of the amphiphilic polymer may have a molar ratio of 10% to 50% with respect to the hydrophilic portion. If it is less than 10%, the polymer may not only partially exist in the aqueous phase, but may also act as a surfactant. If it exceeds 50%, When the hydrophobic part of the molecule is too high to form a complex with the liposome, it may act as an element that destabilizes the liposome structure.

  In one aspect of the present invention, the amphiphilic polymer is produced by polymerization using a general free radical thermal initiation method, and anionic or cationic polymerization is performed to control the molecular weight distribution. May be used. In the case of a natural polymer such as hyaluronic acid, a method in which an alkyl chain is covalently bonded to a side chain may be used in order to add hydrophobicity.

  In one aspect of the present invention, the weight ratio of lipid structure or polymer particle: amphiphilic polymer is 50 to 99% by weight: 1 to 50% by weight, preferably based on the total weight of the mixture. May be 70 to 90% by weight: 10 to 30% by weight, and the lipid structure may contain cholesterol.

  In order to increase the structural stability of the liposome of the drug delivery carrier according to the present invention, when the amphiphilic polymer is further included, the amphiphilic polymer may be a lipid structure or a polymer particle and It may be used in an amount of 1 to 50% by weight, preferably 10 to 30% by weight, based on the total weight of the amphiphilic polymer mixture. This is because, in the case of the content, it is easy to produce the most stable polymer-liposome complex.

  In one aspect of the present invention, the “number average molecular weight” means an average molecular weight obtained by averaging the molecular weights of the component molecular species of the polymer compound having a molecular weight distribution at a fraction or mole fraction. The “weight average molecular weight” may mean an average molecular weight obtained by averaging the molecular weights of the component molecular species of the polymer compound having a molecular weight distribution by the weight fraction. Such a calculation method of the number average molecular weight and the weight average molecular weight may be performed by a method obvious to an ordinary engineer in the technical field of the present invention.

  In one aspect of the present invention, the number average molecular weight or weight average molecular weight of the physiologically active ingredient is 1,000 Da or more, 2,000 Da or more, 3,000 Da or more, 4,000 Da or more, 5,000 Da or more, 6,000 Da or more, 7,000 Da or more, 8,000 Da or more, 9,000 Da or more, 10,000 Da or more, 11,000 Da or more, 12,000 Da or more, 13,000 Da or more, 14,000 Da or more, 15,000 Da or more, 20,000 Da or more, 30,000 Da or more, 50,000 Da or more, 100,000 Da or more, 300,000 Da or more, 500,000 Da, or 1,000,000 Da or more, 5,000,000 Da or less, 4,000,000 Da or less, 3, 1,000,000 Da or less, 2,000,000D Hereinafter, or 1,000,000Da may be less, but is not necessarily limited thereto.

  In one aspect of the present invention, the physiologically active ingredient includes a water-soluble macromolecular substance synthesized as a macromolecule, a macromolecular substance obtained by extraction of a natural product, an enzyme, an EGF (epidermal growth factor), a protein, a peptide, and It may be one or more selected from the group consisting of polysaccharides, and these physiologically active ingredients may exhibit useful effects such as skin moisturizing, whitening and anti-oxidation effects.

  In one aspect of the present invention, the physiologically active ingredient is not particularly limited, but is a water-soluble macromolecule that covers a synthetic, extracted, or naturally-obtained, Sex macromolecules, and such macromolecules may have a beneficial effect on the skin.

  In one aspect, the present invention may relate to a composition comprising a drug delivery carrier according to one aspect of the present invention and a physiologically active ingredient collected inside the carrier.

  In one aspect of the invention, the composition may be a pharmaceutical composition or a cosmetic composition.

  In one aspect of the present invention, the cosmetic composition is not particularly limited in dosage form, and may be appropriately selected according to the intended purpose. For example, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, essence, nutrition essence, pack, soap, cleansing foam, Although it may be manufactured in any one or more dosage forms selected from the group consisting of creasing growth, cleansing cream, body lotion, and body cleanser, it is not necessarily limited thereto.

  When the dosage form of the cosmetic composition according to one aspect of the present invention is a paste, cream, or gel, as a carrier component, animal fiber, plant fiber, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol Silicon, bentonite, silica, talc, or zinc oxide may be used.

  When the dosage form of the cosmetic composition according to one aspect of the present invention is powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder is used as a carrier component. Well, especially if it is a spray, it may further contain a propellant such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether.

  When the dosage form of the cosmetic composition according to one aspect of the present invention is a solution or an emulsion, a solvent, a solvating agent, or an emulsifying agent is used as a carrier component. For example, water, ethanol, There are isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycols, or fatty acid esters of sorbitan.

  When the dosage form of the cosmetic composition according to one aspect of the present invention is a suspension, as a carrier component, water, a liquid diluent such as ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol Suspending agents such as esters and polyoxyethylene sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tragacanth may be used.

  When the dosage form of the cosmetic composition according to one aspect of the present invention is a surfactant-containing cleansing, as a carrier component, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinate monoester, isethionic acid , Imidazolium derivatives, methyl taurates, sarcosinates, fatty acid amide ether sulfates, alkylamide betaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters It's okay.

  The cosmetic composition according to one aspect of the present invention may further contain a functional additive and a component contained in a general cosmetic composition. The functional additive may include a component selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids, and seaweed extracts. The substance is a physiologically active ingredient and may be collected in the drug delivery carrier according to one aspect of the present invention.

  In one aspect of the present invention, the cosmetic composition may further contain a component contained in a general cosmetic composition, if necessary, in addition to the functional additive. Other ingredients included are oil and fat ingredients, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, fungicides, antioxidants, plant extracts , PH adjusters, alcohols, pigments, fragrances, blood circulation promoters, cooling sensates, antiperspirants, purified water, and the like.

  The pharmaceutical composition according to one aspect of the present specification may be in various dosage forms, oral or parenteral. In the case of formulating, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants and the like that are usually used. Solid formulations for oral administration include tablets, pills, powders, granules, soft or hard capsules, and such solid formulations contain at least one excipient for at least one compound, for example, It is prepared by mixing starch, calcium carbonate, sucrose or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to the frequently used simple diluent water and liquid paraffin, various excipients such as wet Agents, sweeteners, fragrances, preservatives and the like may be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, suppositories. As the non-aqueous solvent or suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, or the like may be used. As a suppository base, witepsol, macrogol, tween 61, cacao butter, lauric fat, glycerogelatin and the like may be used.

  In one aspect of the invention, the pharmaceutical dosage form of the composition may also be used in the form of these pharmaceutically acceptable salts and alone or in combination with other pharmaceutically active compounds. It may be used as an appropriate set. The salt is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid, acetic acid, tartaric acid, lactic acid, citric acid. Acid, fumaric acid, maleic acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid and the like may be used.

  In one aspect of the present invention, the composition can be administered parenterally or orally depending on the purpose, so that the daily dose is 0.1 to 500 mg, preferably 1 to 100 mg / kg body weight. It may be administered once to several times. The dose administered to a particular patient may vary depending on the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, and severity of the disease.

  The pharmaceutical composition according to one aspect of the present invention is a powder, granule, tablet, soft or hard capsule, suspension, emulsion, syrup, aerosol or other oral dosage form, ointment, cream, etc. And may be used in any form suitable for pharmaceutical preparations such as suppositories, suppositories, injections and sterile injection solutions, preferably for injections or for external use on the skin. It may be used in the form of an agent.

  The composition according to one aspect of the present invention may be administered to mammals such as rats, mice, domestic animals, and humans from various routes such as parenteral and oral, and all modes of administration can be expected. It may be administered by oral, transdermal, rectal, or intravenous, intramuscular, subcutaneous, uterine lining or intracerebroventricular injection.

  The composition according to one aspect of the present invention may be administered from a variety of routes that can be readily applied by ordinary technicians. In particular, the pharmaceutical composition according to one aspect of the present invention is an external preparation for skin and may be administered from a route applied to the skin surface. The present invention provides an AP-GRR peptide characterized by having a sequence of Gly (Arg) n Gly Tyr Lys Cys (n = 1 to 20) as a cell-penetrating peptide.

  In one aspect of the present invention, in the AP-GRR peptide, the n is 3 to 9.

  In one aspect of the present invention, in the AP-GRR peptide, the sequence is the sequence of SEQ ID NO: 1.

  In one aspect of the present invention, when the drug delivery carrier includes a lipid structure, the drug delivery carrier may further include a stabilizer.

  In one aspect of the present invention, the stabilizer is a cholesterol derivative. Specifically, the stabilizer may be cholesterol. The cholesterol derivative means a derivative having cholesterol as a mother nucleus.

  Specifically, in one aspect of the present invention, the drug delivery carrier may include a lipid structure covalently bound to a cell-penetrating peptide or a peptide chain containing the same; a stabilizer; and a bioactive component.

  In one aspect of the invention, the drug delivery carrier may comprise a lipid structure: stabilizer in a molar ratio of 1-3: 1-2. Specifically, the ratio may be a molar ratio of 1.5 to 3.0: 1.0 to 2.0. More specifically, in the case of a lipid structure, it may consist of a combination of two or more lipids, and at this time, the constituted lipid is included at a lipid molar ratio of 1-2: 1-2. Good.

  In one aspect of the present invention, the lipid structure in the drug delivery carrier comprises one or more of dioleyl phosphatidylethanolamine, phosphatidylcholine, and distearoylphosphatidylethanolamine-polyethylene glycol-maleimide (DSPE-PEG-Mal) complex. May contain.

  In one aspect of the invention, the drug delivery carrier is a dioleyl phosphatidylethanolamine: phosphatidylcholine: cholesterol: distearoylphosphatidylethanolamine-polyethylene glycol-maleimide (DSPE-PEG-Mal) complex 1.0-2.0: It may be included in a molar ratio of 1.0 to 2.0: 1.0 to 3.0: 0.01 to 1.0, specifically, the molar ratio is 1.0 to 1.5: 1. 0 to 1.5: 1.5 to 2.5: 0.1 to 0.3. When the lipid has the kind and molar ratio as described above, a drug delivery carrier that exhibits the most excellent effect in delivering a water-soluble macromolecule into cells can be constructed.

  In one aspect of the present invention, the AP-GRR peptide or peptide chain containing the same covalently bonded to the lipid structure or polymer particle of the drug delivery carrier is 1 mol% based on the lipid structure or polymer particle. Or more, 2 mol% or more, 4 mol% or more, 6 mol% or more, 8 mol% or more, or 10 mol% or more, 15 mol% or less, 10 mol% or less, 8 mol% or less, 6 mol% or less It may be introduced at 4 mol% or less, 2 mol% or less, 1 mol% or less, or 0.1 mol% or less.

  In one aspect of the present invention, the lipid structure is a liposome or an emulsion.

  In one aspect of the present invention, the lipid component of the liposome or emulsion is a phospholipid or a nitrogen lipid having a fatty acid chain having 12 to 24 carbon atoms.

  In one aspect of the present invention, the phospholipids include egg yolk lecithin (phosphatidylcholine), soybean lecithin, lysolecithin, sphingomyelin, phosphatidic acid, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, Diphosphatidylglycerol, cardiolipin, natural phospholipids of plasmalogen, and hydrogenated products, dicetyl phosphate, distearoyl phosphatidylcholine, distearoyl phosphatidyl-ethanolamine that can be obtained from these phospholipids by conventional methods (DSPE), dioleoylphosphatidylethanolamine (DOPE), dipalmitoylphosphatidylcholine, dipalmitoylphosphine Selected from the group consisting of thidylethanolamine, dipalmitoylphosphatidylserine, eleostearoylphosphatidylcholine, eleostearoylphosphatidylethanolamine, synthetic lipids of eleostearoylphosphatidylserine, and fatty acid mixtures obtainable by hydrolysis thereof. It is characterized by more than one kind.

  In one aspect of the present invention, the combination of phospholipids includes a combination of phosphatidylcholine and phosphatidylethanolamine, a combination of phosphatidylcholine and phosphatidylglycerol, a combination of phosphatidylcholine and phosphatidylinositol, a combination of phosphatidylcholine and phosphatidic acid, and phosphatidylcholine. And dioleoylphosphatidylethanolamine, or a combination of phosphatidylcholine, dioleoylphosphatidylethanolamine and phosphatidylserine.

  Specifically, in one aspect of the present invention, the liposome may be a combination of phosphatidylcholine and dioleoylphosphatidylethanolamine.

  In one aspect of the present invention, the combination ratio is characterized in that the mixing ratio of the maximum component to the minimum component is 1: 5 or less.

  In one aspect of the present invention, the combination may be phosphatidylcholine, dioleoylphosphatidylethanolamine, and phosphatidylserine, and the combination ratio of the phosphatidylcholine, dioleoylphosphatidylethanolamine, and phosphatidylserine is 1 It may be ˜4: 1 to 2: 1 to 2.

  In one aspect of the present invention, the lipid component is 0.001 to 20% by weight based on the total weight of the whole liposome dispersion or emulsion.

  In one aspect of the present invention, the polymer particles are compatible with a living body and are degraded in vivo without inducing inflammation or immune reaction, and the degradation product is also harmless in vivo. .

  In one aspect of the present invention, the polymer particles include an amphiphilic polymer or a biodegradable aliphatic polyester polymer.

  In one aspect of the present invention, the polymer particles include a biodegradable aliphatic polyester polymer based on lactic acid and glycolic acid.

  In one aspect of the present invention, the biodegradable aliphatic polyester polymer is poly (D, L-lactic acid) represented by the following chemical formula 3 (poly (D, L-lactic acid)), poly (D L-lactic acid), poly (D-lactic acid), poly (D, L-lactic acid-co-glycolic acid) represented by the following chemical formula 4 (poly (D, L-lactic acid-co-glycolic acid)), Poly (D-lactic acid-co-glycolic acid), poly (L-lactic acid-co-glycolic acid) poly (caprolactone), poly (valerolactone), poly (heath record) City (Poetry) Booty Rate) (poly (hydroxy butyrate)), Poly (hydroxy valerate), poly (1,4-dioxane-2-one) (poly (1,4-dioxane-) 2-one)), poly (ortho ester), and these And wherein the selected from the group consisting of copolymers prepared from dimers is one or more.

(In the above formula, n is an integer of 2 or more.)

(In the above formula, m and n are two or more integers which may be the same or different from each other.)

  In one aspect of the present invention, the biodegradable aliphatic polyester polymer has an average molecular weight of 500 Da to 100,000 Da.

  In one aspect of the present invention, the AP-GRR peptide (A) and the biodegradable aliphatic polyester polymer (B) are configured to be covalently bonded in the AB type or the ABA type. It is characterized by that.

  In one aspect of the present invention, the covalent bond is obtained by adding a base, a linker, or a multiligand compound between an AP-GRR peptide or a peptide chain containing an AP-GRR peptide and the lipid structure or polymer particle. It is characterized by that.

  In one aspect of the present invention, the drug delivery carrier may have an average particle size of 1,000 nm or less. In one aspect of the present invention, the drug delivery carrier has an average particle size of 100 nm or more, 200 nm or more, 300 nm or more, 400 nm or more, 500 nm or more, 600 nm or more, 700 nm or more, 800 nm or more, 900 nm or more, 1,000 nm or more, or 2 3,000 nm or less, 3,000 nm or less, 2,000 nm or less, 1,000 nm or less, 900 nm or less, 800 nm or less, 700 nm or less, 600 nm or less, 500 nm or less, 400 nm or less, 300 nm or less, 200 nm or less, or 100 nm or less It may be.

  In one aspect of the present invention, the drug delivery carrier is contained with a bioactive effectiveness of 0.01 to 30% by weight based on the total weight of the lipid structure, liposome dispersion, emulsion, or polymer particle. A pharmaceutical composition is provided.

  In one aspect of the present invention, the physiologically active ingredient collected in the drug delivery carrier is 0.01 to 30% by weight based on the total weight of the lipid structure, liposome dispersion, emulsion, or polymer particle. Good. Specifically, in one aspect of the present invention, the physiologically active ingredient is 0.01% by weight or more, 0.05% by weight or more, based on the total weight of the lipid structure, liposome dispersion, emulsion, or polymer particle. 0.1% by weight or more, 0.5% by weight or more, 1% by weight or more, 2% by weight or more, 3% by weight or more, 4% by weight or more, 6% by weight or more, 10% by weight or more, 15% % By weight or more, 20% by weight or more, 25% by weight or more, or 30% by weight or more, 30% by weight or less, 25% by weight or less, 20% by weight or less, 15% by weight or less, 10% by weight or less, 6% by weight % Or less, 5% or less, 4% or less, 3% or less, 2% or less, 1% or less, 0.5% or less, 0.1% or less, 0.05% or less, or It may be 0.01% by weight or less.

  In one aspect of the present invention, the composition has a dosage form selected from the group consisting of an external preparation for skin, an oral preparation, and an injection.

  The present invention is a newly devised GRR peptide having excellent biomembrane permeability, which is rich in arginine and has Glycine (Gly) amino acid and Glycine (Gly) -Tyrosine (Tyr) -Lycine (both ends). (Lys) -Cystein (Cys) A peptide having an amino acid chain, a liposome having a structure for collecting water-insoluble or water-soluble macromolecules such as drugs, genes, oligopeptides and proteins, polymer nanoparticles, phospholipid-polymer This is to improve the bioavailability when the substance to be delivered is delivered by various routes such as transdermal, oral, injection, etc., by modifying the surface of a delivery carrier such as a complex or emulsion. . In the AP-GRR peptide, the number of Arg in the sequence of Gly (Arg) n Gly Tyr Lys Cys includes 1 to 20, preferably 3 to 9. This is because it has high delivery efficiency and is easy to manufacture in combination with a drug delivery carrier.

  In one aspect of the present invention, the AP-GRR peptide or the peptide chain containing AP-GRR is not particularly limited, and examples thereof include amide 4-methylbenzylidylamine hydrochloride (MBHA) resin and ABI 433 synthesis. Using an instrument, it may be synthesized by solid phase peptide synthesis (SPPS) according to the Fmoc (N- (9-flurenyl) methoxycarbonyl) / t-butyl method (M. Bodansky, A. Bodansky, The Practice of Peptide Synthesis; Springer: Berlin, Heidelberg, 1984, JM Stewart, JD Young, Solid Phase Peptide Synthesis, 2nd ed; Pierce Chemical Co: Rockford. IL, 1984).

  In one aspect of the present invention, the drug delivery carrier may use a structure such as a lipid structure, a liposome, an emulsion, or a polymer particle. In one aspect of the present invention, phospholipids having a fatty acid chain having 12 to 24 carbon atoms or nitrogen lipids may be used as the lipid component of the lipid structure during the production of the liposome or emulsion. This is because they are easy to use as a component of lipid-based drug delivery carriers that can be used in pharmaceutical compositions such as external preparations for skin, oral preparations, and injections.

  In one aspect of the present invention, generally, phospholipids are more preferable as the lipid component of the lipid structure. For example, egg yolk lecithin (phosphatidylcholine), soybean lecithin, lysolecithin, sphingomyelin, phosphatidic acid, Natural phospholipids such as phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, diphosphatidylglycerol, cardiolipin, plasmalogen, hydrogenated products and dihydrides that can be obtained in the usual way from these phospholipids Cetyl phosphate, distearoyl phosphatidylcholine, distearoyl phosphatidyl-ethanolamine, dioleoylphosphatidylethanolamine, dipalmitoylphos Synthetic lipids such as watidylcholine, dipalmitoyl phosphatidylethanolamine, dipalmitoyl phosphatidylserine, eleostearoylphosphatidylcholine, eleostearoylphosphatidylethanolamine, eleostearoylphosphatidylserine, and fatty acid mixtures obtainable by hydrolysis thereof are preferred. .

  In one aspect of the present invention, lipids may be used alone or in combination of two or more. When a mixture of two phospholipids is used, preferably a combination of phosphatidylcholine: phosphatidylethanolamine, phosphatidylcholine: phosphatidylglycerol, phosphatidylcholine: phosphatidylinositol, phosphatidylcholine: phosphatidic acid, phosphatidylcholine: dioleoylphosphatidylethanolamine, etc. May be used. The mixing ratio varies depending on the composition of the components to be mixed, but the mixing ratio of the maximum component to the minimum component is preferably 1: 5 or less. This is because it is easy to produce a lipid-based drug delivery carrier by mixing two or more phospholipids. For example, in the case of phosphatidylcholine: dioleoylphosphatidylethanolamine, 1: 1 or 2: 1, 3: 1, 4: 1, 5: 1, 1: 5, 1: 4, 1: 3, 1: 2, etc., respectively. It can be produced with various molar ratios. Even when three types of phospholipids are mixed and used, for example, phosphatidylcholine: dioleoylphosphatidylethanolamine: phosphatidylserine is 1: 1: 1 or 2: 1: 1, 3: 1: 2, 3: It can be produced at a molar ratio of 2: 1.3: 2: 2.4: 1: 1, 4: 2: 1, and various mixing ratios.

  In one aspect of the invention, the lipid component of the drug delivery carrier is in an amount of 0.001-20 wt%, preferably 0.2-10 wt%, based on the total weight of the total lipid structure, liposome dispersion or emulsion. Used in. This is because, in the case of the content, it is easy to produce a drug delivery carrier.

  In producing the polymer particles according to one aspect of the present invention, it is compatible with the living body and not only decomposes in vivo without inducing inflammation, immune reaction, etc., but also its degradation products also The polymer must be harmless in vivo. Biodegradable aliphatic polyester polymers based on lactic acid and glycolic acid as polymers that satisfy these conditions are the most widely used polymers approved by the US Food and Drug Administration (FDA). Yes. Typical examples thereof include poly (D, L-lactic acid) (poly (D, L-lactic acid)), poly (L-lactic acid), poly (D-lactic acid) and the above chemical formula. 4 (poly (D, L-lactic acid-co-glycolic acid)), poly (D-lactic acid-co-glycolic acid), poly (L -Lactic acid-co-glycolic acid), and poly (caprolactone) (poly (caprolactone)), poly (valerolactone), poly (hydroxy butyrate), poly (hydroxy (Poly (hydroxy valerate)), poly (1,4-dioxane-2-one) (poly (1,4-dioxane-2-one)), poly (ortho ester) And copolymers produced from these monomers.

In one aspect of the present invention, the molecular weight of the biodegradable aliphatic polyester polymer is not particularly limited, but when it is less than 500 or more than 100,000, structural instability as a drug delivery carrier increases. Therefore, the weight average molecular weight may be 500 to 100,000, preferably 5,000 to 50,000.
In one aspect of the present invention, the molecular weight of the biodegradable aliphatic polyester polymer is 1,000 Da or more, 2,000 Da or more, 3,000 Da or more, 4,000 Da or more, 5,000 Da or more, 6,000 Da or more. 7,000 Da or more, 8,000 Da or more, 9,000 Da or more, 10,000 Da or more, 11,000 Da or more, 12,000 Da or more, 13,000 Da or more, 14,000 Da or more, 15,000 Da or more, 20,000 Da or more 30,000 Da or more, 50,000 Da or more, or 100,000 Da or more, 200,000 Da or less, 150,000 Da or less, 100,000 Da or less, 90,000 Da or less, 80,000 Da or less, 70,000 Da or less, 60,000 Da or less, 50,000 Da Below 40,000 Da or less, 30,000 Da or less, 20,000 Da or less, 10,000 Da or less, 5,000 Da or less, 3,000 Da or less, or 1,000 Da or less, but not necessarily limited thereto. It is not a thing.

  In one aspect of the present invention, in the case of poly (D, L-lactic acid-co-glycolic acid) represented by Chemical Formula 4, the ratio of lactic acid to glycolic acid monomer is adjusted, or a polymer synthesis process is performed. By changing it, biodegradable polymers having various degradation lifetimes can be obtained. Such biodegradable aliphatic polyester-based polymers have long been used as drug delivery carriers or surgical sutures, and their biocompatibility has already been proven.

  In one aspect of the present invention, the AP-GRR peptide (A) and the biodegradable aliphatic polyester polymer (B) are not particularly limited, but are AB type or ABA type. It may consist of This is because the carboxyl group and hydroxyl group present at both ends of the biodegradable aliphatic polyester polymer are substituted with other functional groups so as to facilitate covalent bonding, and the polymer is replaced. The terminal can be obtained by reacting with the terminal group of the AP-GRR peptide having a group that easily reacts with the terminal group or the terminal group of the peptide chain containing the AP-GRR peptide. For example, the covalent bond between the AP-GRR peptide or the peptide chain containing the AP-GRR peptide and poly (D, L-lactic acid-co-glycolic acid) is poly (D, L-lactic acid-) substituted with maleimide. co-glycolic acid) and an AP-GRR peptide having a terminal group substituted with a thiol may be synthesized by a covalent bond.

  In one aspect of the present invention, the covalent bond is not particularly limited, but may be separately provided between an AP-GRR peptide or a peptide chain containing the AP-GRR peptide and the lipid structure, liposome, emulsion, or polymer. Or a base or a linker or a multiligand compound.

  In one aspect of the present invention, the physiologically active component collected in the internal structure of the drug delivery carrier is not limited in its type, such as water-soluble or water-insoluble, as long as it can be applied to a living body. For example, it may be a single component, an animal or plant or fungus extract, and a mixture containing two or more of these, and may be used by adjusting according to the purpose and circumstances. Ingredients having the effect of whitening enhancement or wrinkle and anti-aging and treatment may be used. These physiologically active ingredients are incorporated in an amount of 0.01 to 30% by weight, preferably 0.1 to 20% by weight, based on the total weight of the lipid structure, liposome dispersion, emulsion or polymer particle. It may be. This is because the composition has a content that is easy to form, such as an external preparation for skin, an oral preparation, and an injection.

  The average particle size of the drug delivery carrier manufactured using the AP-GRR peptide according to one aspect of the present invention is preferably as small as possible, and when considering the stability of the colloid, it is at least 1,000 nm or less, preferably 500 nm or less. It is.

  The method for producing liposomes, emulsion polymer particles and the like presented in the present invention is not particularly limited. For example, they may be produced by the following method.

  As a method of forming a drug delivery carrier in which a physiologically active ingredient is incorporated using the lipid component presented in the present invention, the phospholipids and the stabilizer are dissolved in an organic solvent, and then the solvent is evaporated and depressurized. To form a lipid film, add an aqueous solution, and then apply ultrasonic waves, disperse phospholipids and stabilizers dissolved in an organic solvent in an aqueous solution, and then apply ultrasonic waves, stabilize phospholipids After dispersing or dissolving the agent in an organic solvent, the organic solvent is extracted or evaporated with an excessive amount of water. After dispersing or dissolving the phospholipids and stabilizer in the organic solvent, a homogenizer or high-pressure emulsifier is used. A method of evaporating the solvent by vigorous stirring, a method in which phospholipids and stabilizers are dispersed or dissolved in an organic solvent and then dialyzed with an excessive amount of water, and a phospholipid and stabilizers are dispersed in an organic solvent. Or after dissolving Although there is a method of gradually adding water, it not necessarily limited thereto.

  In relation to the production method, in order to dissolve the phospholipids and the stabilizer in the organic solvent, mechanical force is applied or heating is performed at 20 to 100 ° C., preferably 70 ° C. or less.

  When the physiologically active ingredient is water-soluble, the physiologically active ingredient is dissolved in water or dissolved in an aqueous solution, and administered together at the stage of adding the aqueous solution or water. When the physiologically active ingredient is insoluble in water, the physiologically active ingredient can be dissolved in an organic solvent and then added to the organic solvent containing the lipid component by the method described above.

  Organic solvents that can be used to dissolve phospholipids and stabilizers or water-insoluble physiologically active ingredients in the above method include acetone, dimethyl sulfoxide, dimethylformamide, N-methylpyrrolidone, dioxane, tetrahydrofuran, acetic acid, ethyl A solvent selected from acetate, acetonitrile, methyl ethyl ketone, methylene chloride, chloroform, methanol, ethanol, ethyl ether, diethyl ether, hexane, and petroleum ether or a mixture of these may be used, but is not necessarily limited thereto. It is not a thing.

  In addition, as a method for producing polymer particles presented in the present invention, a method of adding ultrasonic waves after dispersing a polymer as it is in an aqueous solution, an organic solvent with an excessive amount of water after dispersing or dissolving the polymer in an organic solvent Extract or evaporate, disperse or dissolve the polymer in an organic solvent, then vigorously stir using a homogenizer or high-pressure emulsifier to evaporate the solvent, after disperse or dissolve the polymer in an organic solvent Examples include a method of dialysis with an excessive amount of water, a method of gradually adding water after dispersing or dissolving a polymer in an organic solvent, and a method of manufacturing using a supercritical fluid (T. Niwa et al., J. Pharm. Sci. (1994) 83, 5, 727-732; CS Cho et al., Biomaterials (1997) 18, 323-326; T. Govender et al., J. Control. Rel. (1999) 57 171-185; MF Zambaux et al., J. Control. Rel. (1998) 50, 31-40).

  Examples of the organic solvent that may be used for the production of the polymer particles of the present invention include acetone, dimethyl sulfoxide, dimethylformamide, N-methylpyrrolidone, dioxane, tetrahydrofuran, ethyl acetate, acetonitrile, methyl ethyl ketone, methylene chloride, chloroform, methanol, ethanol. , Ethyl ether, diethyl ether, hexane, or petroleum ether, but is not necessarily limited thereto. At this time, the solvent may be used alone, or two or more solvents may be mixed and used.

  The distearoylphosphatidylethanolamine-polyethylene glycol-maleimide (DSPE-PEG-Mal) complex used in the present specification may have a structure represented by the following chemical formula 5.

  Hereinafter, the present invention will be described more specifically with reference to examples, comparative examples, and experimental examples. The materials, reagents, costs, operations, and the like shown in the following examples can be changed as appropriate without departing from the spirit of the present invention. Therefore, the scope of the present invention is not limited by the specific examples shown below.

[Examples and Comparative Examples] Production of Drug Delivery Carriers Peptide chains containing AP-GRR peptides were prepared using Fmoc (N- (N- (4)) using amide 4-methylbenzylidylamine hydrochloride (MBHA) resin and ABI 433 synthesizer. 9-Furenyl) methoxycarbonyl) / t-butyl method, synthesized by solid phase peptide synthesis (SPPS), and purified by 90% purity using Reverse High Performance Liquid Chromatography Purified as above. As a result of analyzing this with a mass spectrometer (Agilent 1100 series), the success or failure of the synthesis was confirmed by measuring the molecular weight. This confirmed that a peptide having the amino acid sequence of AP-GRR SEQ ID NO: 1 was synthesized. The lipid structure production process in which the AP-GRR is introduced on the surface is as follows.

  The lipids listed in the lipid composition of Table 1 were mixed according to the molar ratio of the lipids, then dissolved in an organic solvent mixed with chloroform: methanol (95: 5, volume / volume), and then the solvent was evaporated under reduced pressure. To form a film. For example, in the case of Example 2, dioleyl phosphatidylethanolamine: phosphatidylcholine: cholesterol: distearoylphosphatidylethanolamine-polyethylene glycol-maleimide complex was mixed in a molar ratio of 1.5: 1.1: 2: 0.4. Then, after dissolving in chloroform: metalol (95: 5, volume / volume) mixed organic solvent, the solvent was evaporated under reduced pressure to form a film.

  Rhodamine B (manufactured by Sigma-Aldrich, CAS NO. 81-88-9) or dextran, which is a fluorescent labeling substance dissolved in PBS (manufactured by WELGENE, hereinafter the same), is removed from the film from which the organic solvent has been removed. After adding (dextran) -RITC (number average molecular weight of about 10,000 Da) (manufactured by Sigma-Aldrich), ultrasonic waves were added to produce a liposome lipid dispersion. AP-GRR peptide was dissolved in PBS and added to the liposome lipid dispersion produced as described above, and then stirred at room temperature (about 25 ° C.) for reaction. Through such a reaction, a thiol group of AP-GRR and a maleimide functional group derived on the surface of the liposome are reacted for several minutes to produce a liposome into which AP-GRR is introduced, and such a bond The reaction is known to be extremely stable. The concentration of lipid in the final aqueous solution is 0.2 wt%.

  According to the said method, the Example and the comparative example were manufactured according to the lipid composition and the molar ratio of a lipid as shown in Table 1 below. Examples 1 to 6 in the following table are liposomes into which AP-GRR was introduced, provided that Example 1 did not carry a fluorescent substance. Comparative Example 1 is a liposome into which AP-GRR has not been introduced and does not carry a fluorescent substance. Comparative Examples 2 and 4 correspond to PBS solutions in which a fluorescent substance is dissolved. Comparative Examples 3 and 5 Is a liposome into which AP-GRR has not been introduced, and contains a fluorescent substance. DSPE-PEG-Mal was calculated based on the molecular weight of DSPE minus the molecular weight of PEG and Mal in the molar ratio of lipids of Examples 1 to 5, and calculated only by the lipid ratio to calculate AP-GRR. Introduced at 4 mol% and 8 mol% in the lipids constituting Further, in order to conduct an experiment accompanying the introduction of the amphiphilic polymer, as Example 6, an example in which only the amphiphilic polymer was further added to the composition of Example 5 was set. The poly (hydroxyethyl methacrylate-co-stearyl methacrylate) copolymer used in Example 6 corresponds to a number average molecular weight of 47,552 (PDI 2.20).

  The polymerization method of such an amphiphilic polymer is obtained by adding 0.0995654: 0.232315 monomers of hydroxyethyl methacrylate (purchased from Sigma-Aldrich) and stearyl methacrylate (purchased from Sigma-Aldrich) to 150 g of ethanol. A molar ratio was added, and 0.003319 mol of AIBN (Azobisisobutyronitrile) (purchased from JUNSEI), which is a radical polymerization initiator, was added and stirred at 75 ° C. overnight to perform polymer polymerization. After polymerization, the heating is interrupted and allowed to stand until the temperature drops to room temperature. Thereafter, the mixture was stirred while slowly adding 5-fold to 10-fold ether based on the ethanol solution, and the solvent was removed with a filter to obtain a precipitate produced by stirring. The precipitate thus obtained was vacuum-dried to obtain 40 g of poly (hydroxyethyl methacrylate-co-stearyl methacrylate) copolymer powder.

  In the production of Example 6, the amphiphilic polymer was produced in the same process except that it was added together when the lipid was dissolved in the organic solvent.

  * DOPE: Phosphatidylethanolamine (obtained from Doosan Biotech), PC: Phosphatidylcholine (LIPOID), Chol: Cholesterol (Sigma-Aldrich), DSPE-PEG-Mal (Molecular weight 2941.605) (NOF) ): Distearyl phosphatidylethanolamine-polyethylene glycol-maleimide complex (N-[(3-Maleimide-1-oxopropyl) aminopropyl polyethyleneglycol-carbamyl] distearoylphosphatidyl-ethanolamine)

[Production of Examples Mixed with o / w Nanoemulsion]
In order to compare the absorption capacity and the particle size analysis for the skin when mixed with an o / w nanoemulsion in the drug delivery carrier accompanying the introduction of the amphiphilic polymer, the drug delivery carriers of Example 5 and Example 6 were used. Examples 7 and 8 were prepared as experimental samples mixed with o / w nanoemulsion.

  Specifically, an aqueous phase and an oil phase were prepared as shown in Table 2 below.

  The aqueous phase and the oil phase produced by the above composition were separately heated and dissolved at 70 ° C., and the oil phase was slowly added to the aqueous phase, and a homomixer (TK Homomixer Mark II, Takushu Kika Kogyo Ltd. , Japan) for 3 minutes. The o / w emulsion thus obtained by emulsification was further processed at 1000 bar for 3 cycles in a high-pressure emulsifier (Microfluidics Corp., USA) to obtain a nanoemulsion having an average particle size of about 150 nm. The nanoemulsions of Examples 5 and 6 were mixed with the nanoemulsions thus prepared at a weight ratio of 1: 1 by azimixing. Thus, compositions in which the drug delivery carriers of Example 5 and Example 6 and the nanoemulsion were mixed were prepared, and these were set as Example 7 and Example 8, respectively.

[Test Example 1] Particle size analysis of drug delivery carrier and measurement of surface potential For the liposome solutions of Examples 1 to 8 and Comparative Examples 1, 3, and 5, particle size analysis and surface potential were measured with a Malvern Zetasizer. The measurement results are shown in Table 3 below.

  According to the results of Table 3, it was confirmed that a comparatively uniform sample having an average particle size of about 100 nm was produced in both the comparative example and the example.

  Moreover, when AP-GRR was introduce | transduced, it has confirmed that the surface potential changed from a negative value to a positive value compared with the lipid structure which is not so. In addition, in Examples 2 and 4 in which the amount of AP-GRR introduced was as high as 8 mol%, the surface potential had a higher positive surface potential than in Examples 3 and 5 in which the amount of AP-GRR introduced was 4 mol%. I was able to confirm. From these results, it was confirmed that AP-GRR was introduced into the drug delivery carrier of the present invention and its physical properties were changed.

  Also, looking at the results of the particle size analysis in the nanoemulsion state, Example 8 in which the amphiphilic polymer was introduced was smaller in average particle size and higher in particle size stability than Example 7 that was not. You can confirm the fact. From these results, it can be concluded that in Example 7, the particle size increased and the PDI value increased while the recombination of the new emulsion occurred between the liposome containing the cell-permeable peptide and the nanoemulsion. This is attributed to structural instability in the emulsifier form, which is one of the disadvantages of liposome formulations indirectly. However, in the case of Example 8, it is judged that the amphiphilic polymer plays a role of a protective colloid while supporting the double lipid layer of the liposome, and as a result, the particle size result is similar to Example 6 produced with the lipid structure. No new high particle size particles were observed.

[Test Example 2] Experiment of TEM structure analysis of drug delivery carrier For Comparative Example 1 and Example 1, the structure was confirmed using a transmission electron microscope (Libra 120, Carl Zeiss, acceleration voltage 120 kV). The results are shown in FIG.
As shown in FIG. 1, the liposome structure of Example 1, which is a drug delivery carrier corresponding to a liposome into which AP-GRR was introduced, was slightly larger than that of Comparative Example 1 in which AP-GRR was not introduced. I was able to confirm. Moreover, although AP-GRR was introduce | transduced, it can confirm that the structure of a liposome is similar in the comparative example 1 and Example 1, and from this, the macromolecule of this invention can be carry | supported. It was confirmed that the drug delivery carrier having a structure was structurally similar to general liposomes. However, according to the following experimental examples, the drug delivery carrier of the present invention is structurally similar to general liposomes, but has the effect of easily transferring water-soluble macromolecules into cells.

[Test Example 3] Evaluation of intracellular delivery ability of drug delivery carrier by cell-containing analysis For Examples 2-5 and Comparative Examples 2-5, FACS analysis was performed to confirm the intracellular delivery ability of the drug delivery carrier. Carried out. The liposome system of Examples and Comparative Examples was added to pre-cultured HaCaT cells (obtained from CLS cell line service), cultured for 4 hours at 37 degrees Celsius, and then cells of each sample group were collected and dispersed in PBS solution FACS analysis was performed. For FACS analysis, BD FACScalibur equipment (Beckton Dickinson Bioscience, San Jose, Calif.) Was used, and red fluorescence in 10,000 HaCaT cells was measured for each experimental group, and a sample was obtained with CellQuest software. analyzed. Thereby, the amount of rhodamine B taken up into the cells was quantitatively compared and analyzed, and the results are shown in FIG. FIG. 2A shows the results for Rhodamine B, the FACS analysis graph results for Examples 2 and 3 and Comparative Examples 2 and 3, and FIG. 2B shows the results for dextran-RITC, The result with respect to the comparative examples 4 and 5 is shown. In the case of FIG. 2C, the result of the graph is numerically shown, and corresponds to a value indicating mean values (mean) and standard deviations (SD). The y-axis in the graph means the number of cells, and the x-axis means the amount delivered into the cells.

  According to FIG. 2A and FIG. 2C, the graph corresponds to the graphs of Example 3, Example 2, Comparative Example 3, and Comparative Example 2 in descending order of the average value from the right side, and in the case of the example in which AP-GRR is introduced. It was confirmed that the average amount delivered into the cells was larger than that of the comparative example that was not so. Therefore, in the case of intracellular delivery ability, the treatment with liposome is higher than the PBS solution, and the drug delivery carrier of the present invention in which AP-GRR is introduced into the liposome is superior to the single liposome. I was able to confirm. From this, in the case of the drug delivery carrier according to the present invention, it was confirmed that it was easy to deliver a water-soluble low-molecular substance such as rhodamine B into the cells. Moreover, when Examples 2 and 3 having different AP-GRR introduction amounts were compared, the case of Example 2 introduced at 8 mol% into the lipid constituting the liposome was more average than Example 3 in which 4 mol% was introduced. It was confirmed that the value was small. There is no significant difference in constructing the drug delivery carrier, but it is judged that 4 mol% is more preferable for the amount of AP-GRR introduced.

  According to FIG. 2B and FIG. 2C, the graph corresponds to the graphs of Example 5, Example 4, Comparative Example 5, and Comparative Example 4 in descending order of the average value from the right side, and in the case of the example in which AP-GRR is introduced. It was confirmed that the average amount delivered into the cells was larger than that of the comparative examples that were not so. Therefore, in the case of intracellular delivery ability, the treatment with liposome is higher than the PBS solution, and the drug delivery carrier of the present invention in which AP-GRR is introduced into the liposome is superior in delivery ability to a single liposome. I was able to confirm. Further, in the case of Examples 4 and 5, unlike those collected in Examples 2 and 3, the polymer is dextran-RITC and has a molecular weight different by about 20 times. The tendency to be delivered intracellularly is similar to the results when using rhodamine B. From this, it was confirmed that the drug delivery carrier of the present invention was excellent in intracellular delivery ability even in the case of a water-soluble macromolecule. Further, when Examples 4 and 5 having different AP-GRR introduction amounts are compared, it can be confirmed that the average value is larger in Example 5 in which 4 mol% is introduced, as in the case of Rhodamine B. did it.

[Test Example 4] Evaluation of cell transfer rate using immunofluorescence staining and confocal laser microscope DMEM medium (LONZA) supplemented with 10 wt% FBS (GIBCO) and 100 IU penicillin G (LONZA) Then, HaCaT cells (obtained from CLS cell line service) were dispensed into 8 well chamber slides at 25,000 cells / well. Next, the wells were washed using PBS (phosphate buffered saline), and then diluted with a control group in which nothing was added, the above Examples and Comparative Examples, and treated for 3 hours. Immunofluorescence staining (Immunofluorescence; IF) was performed on the treated cells. Each well was washed using PBS in which 1 mM CaCl ₂ and 1 mM MgCl ₂ were incorporated (hereinafter, the PBS in this test example was the same) to wash the cells contained in the well. The cells were reacted and fixed at room temperature for 10 minutes using 3.5% by weight paraformaldehyde. The fixed cells were washed again 3 times for 10 minutes using PBS. Thereafter, 0.1% Triton X-100 was treated for 5 minutes. After the treatment, it was washed again with PBS for 10 minutes three times. To stain the nuclei, the cells were stained for about 3 minutes using Propidium iodide (PI). PBST again (prepared by mixing 0.05 wt% Tween 20 in PBS, and obtained from SIGMA in the case of Tween 20. PBS for the manufacture of PBST does not contain calcium chloride or magnesium chloride.) After washing three times using a mounting, a mounting solution was added to cover the coverglass. The slides thus stained were photographed using a confocal laser scanning microscope (Confocal microscopy, Ziess). Such results are shown in FIGS. FIG. 3 shows the results for liposomes containing water-soluble low molecular weight rhodamine-B as a fluorescent substance, and FIG. 4 shows the results for liposomes containing dextran-RITC, which is a water-soluble macromolecule, as a fluorescent substance. The part stained red in the photograph shows rhodamine-B or dextran-RITC, and the part stained blue shows the nucleus.

  According to FIG. 3, the cell transfer rate of Comparative Example 3 which is a single liposome is slightly higher than that of Comparative Example 2 which is a PBS solution, and the AP-GRR introduced into this is compared with such a liposome. It was confirmed that the cell transfer rate was significantly higher in the case of the drug delivery carrier of the invention.

  According to FIG. 4, a result similar to rhodamine-B could be confirmed even in the case of dextran-RITC having a molecular weight about 20 times larger. From the photographs of Comparative Examples 4 and 5, it can be confirmed that dextran-RITC is hardly taken up into the cells. On the other hand, from the photographs of the results of Examples 4 and 5, the red fluorescent substance is found. It was confirmed that a large amount was taken up into the cell, and from this, it was confirmed that the drug delivery carrier of the present invention showed a remarkable effect in delivering the water-soluble macromolecule into the cell.

[Test Example 5] Evaluation of skin stability In order to confirm the skin stability of Examples 1 to 5 and Comparative Examples 1 to 5, subjects were 18 adult women and 12 men (average age 32.5 years). ) And a patch test in which the products containing the liposomes of Examples and Comparative Examples were applied, and the skin stability of the composition of the present invention was evaluated. In the measurement method, 28 hours after the patch was attached, the patch was removed, the first observation was performed 30 minutes later, and the second observation was performed after 96 hours had elapsed. In order to examine the strength related to skin irritation of the composition, weight was given according to the degree of positive reaction of the skin, and the skin irritation of the composition was visually determined by obtaining the skin average reactivity. The results are shown in Table 4 below.

  According to the results of Table 4 above, it was confirmed that both the examples and the comparative examples did not irritate the skin when they were produced by being included in the composition. Therefore, the cosmetic composition containing the liposome of the present invention could be judged to be excellent in stability to the skin.

[Test Example 6] Evaluation of qualitative effects of drug delivery carriers by skin absorption experiment Skin absorption experiments were conducted for Comparative Examples 2 to 5, Example 3, Example 5, Example 7, and Example 8. For skin absorption experiments, use the skin extracted from the back of the ears, which are a by-product of slaughter pigs (obtain the ears of pigs that have not been depilated at the slaughterhouse, and use them in the experiment), and wash the extracted skin And included in the comparative examples and examples over a period of 4 hours and 18 hours in a Franz-cell instrument (Franz-type vertical diffusion cell system (Microette Plus Auto Sampling System, Hanson Research, USA)), respectively. The skin absorption experiment of the fluorescent substance was conducted.

  Porcine ear skin that had been absorbed with Franz Cell was placed in Mold and covered with OCT compound (# 4583, SAKURA Tissue-Tek, USA) to prevent bubbles from forming. Thereafter, it was rapidly frozen at −196 ° C. using liquid nitrogen. The rapidly frozen pig ear skin was sliced at a thickness of 6 μm by using a cryosection machine (CM1950 cryostat, Leica, Germany), and was pasted on a silane-coated slide glass. Such a slide glass was dried for 10 minutes in a light-shielded place at room temperature (25 ° C.) and then observed using a fluorescence microscope (BX53, Olympus, Japan). The images were observed under the same fluorescence intensity and exposure time, and representative images were taken using a cooled digital color camera (DP72, Olympus, Japan). The results are shown in FIGS. The scale bar shown in white in FIGS. 5 and 6 corresponds to 500 μm. In the case of FIG. 5, the exposure time is set to 256 ms, and in the case of FIG. 6, the exposure time is set to 64 ms. In the case of fluorescence intensity, a mercury lamp (U-HGLGPS, olympus, japan) is used as a light source, and in the case of fluorescence intensity, values (0, 3, 6, 12, Analysis was performed in accordance with 6 out of 25, 50, and 100).

  According to FIG. 5, in the case of rhodamine B, it is a substance having a relatively small molecular weight, and even when dissolved in PBS as in Comparative Example 2, when absorbed for 18 hours, it is absorbed into the skin more than a certain amount, When applied to general liposomes as in Comparative Example 3, it can be confirmed that transdermal absorption occurs from the case of absorption for 4 hours. This is considered that absorption of a water-soluble component increased by the disturbance effect | action of the stratum corneum intercellular lipid by a liposome. In addition, in the case of dextran-RITC corresponding to a polymer substance, it was confirmed that in any of Comparative Examples 4 and 5, absorption into the skin was hardly caused.

  According to FIG. 6, in the case of rhodamine B, it is confirmed that the absorption of rhodamine B is remarkably exhibited even after absorption for 4 hours, compared with the case of the drug delivery carrier according to one aspect of the present invention dissolved in PBS. can do. Compared to the general liposome as Comparative Example 3 in FIG. 5, the absorption of Rhodamine B applied to the drug delivery carrier of Example 3 was formed with a wider fluorescence range and a larger thickness. It can be confirmed that a more excellent effect is exhibited. Such effects are considered to be due to the fact that the basic stratum corneum lipid perturbation effect of liposomes and the positive charge effect due to the polyarginine group of the cell-penetrating peptide work effectively for absorption into the skin. .

  Further, in the case of Comparative Examples 4 and 5, in the case of Example 5 applied to the drug delivery carrier according to one aspect of the present invention even in the case of dextran-RITC in which absorption to the skin hardly occurs, It was possible to confirm that absorption occurred. From these results, it is confirmed that the drug delivery carrier according to one aspect of the present invention exhibits a heterogeneous and remarkable effect of delivering a polymer substance having a molecular weight reaching the level of 10,000 Da into the skin. be able to.

Test Example 7 Evaluation of Quantitative Effect of Drug Delivery Carrier by Skin Absorption Experiment The stratum corneum was compared to the pig ear skin absorbed in the Franz cell device in Test Example 6 for 4 hours with the Franz cell device. The skin tissue excluding the stratum corneum and the receptor sample were prepared as follows. In the case of the stratum corneum sample (SC), the surface of the skin tissue is subjected to 3M tape stripping 3 times after 6 mm biopsy on the absorbed pig ear skin, and the tape is mixed with water and methanol at 1: 1. Extracted with 6 ml of solvent mixed in proportion. In the case of a skin tissue sample excluding the stratum corneum (Skin without SC), it was a tissue sample obtained after tape stripping, and extracted with 2 ml of a solvent in which water and methanol were mixed at a ratio of 1: 1. In the case of a receptor sample, it was obtained by dispensing 1 ml of a PBS solution in the receptor part after an absorption experiment.

  Each sample thus obtained was analyzed with a spectrophotometer (Spectrophotometer, F4500, HITACHI), and the analysis conditions were as follows.

Analysis conditions:
-Ex slit (excitation slit): 2.5 / emi slit (emission slit): 2.5
-Exi (wavelength of light for excitation): 554nm / emi (wavelength of light for excitation): 579nm

  This analysis corresponds to a result obtained by obtaining an emission result of a fluorescent substance after irradiation with light having a wavelength of 544 nm, and detecting and calculating an intensity value corresponding to 579 nm therefrom. Quantitative results of the amount of absorption of the fluorescent material obtained under such analysis conditions are shown in FIGS. 7 and 8 and Table 5 below. As a result of the analysis, no fluorescence was detected from the receptor portion.

  According to the results of FIGS. 7 and 8 and Table 5, in the case of Comparative Example 2, Comparative Example 3 and Example 3 containing Rhodamine B, in the skin tissue excluding the stratum corneum and the stratum corneum, Comparative Example 2 <Comparative Example It was confirmed that the concentration of rhodamine B absorbed in the order of 3 <Example 3 increased. From this, the degree of skin absorption enhancement effect by the liposome structure in the case of a substance having a low molecular weight can be confirmed.

  In the case of Comparative Example 4, Comparative Example 5 and Example 5 containing dextran-RITC, a slightly different aspect from Rhodamine B was shown. In the case of the fluorescent substance absorbed in the stratum corneum, it was hardly absorbed in Comparative Example 4, and no absorbed fluorescent substance was observed in Comparative Example 5 applied to general liposomes. On the other hand, in the case of Example 5 applied to the drug delivery carrier according to one aspect of the present invention, a significantly higher absorption effect than Comparative Examples 4 and 5 could be confirmed. Even in the skin tissue excluding the stratum corneum, in the case of Example 5, a significantly high fluorescent substance absorption could be confirmed, whereas in Comparative Examples 4 and 5, a conspicuous skin absorption effect was not seen. Therefore, according to such a result, in the case of the drug delivery carrier according to one aspect of the present invention, it can be confirmed that even a substance having a high molecular weight exhibits a high absorption effect on the stratum corneum and skin tissue. Such an effect is a remarkable and heterogeneous effect as compared with existing liposomes.

  In addition, when mixed with an emulsifier form in the form of a nanoemulsion, when the drug delivery carrier according to one aspect of the present invention contains an amphiphilic polymer, the skin absorption rate is significantly higher than when it is not. It was confirmed from the results of Example 7 and Example 8. Specifically, when Example 6 which is a drug delivery carrier containing an amphiphilic polymer in nanoemulsion (emulsifier form) and Example 5 not containing this are mixed, Example 8 containing an amphiphilic polymer is mixed. Compared to Example 7, it was confirmed that the amount of skin absorbed by dextran-RITC, which is a macromolecular bioactive component, was 7 times or more higher. This is considered to be an increase in skin absorption accompanying an increase in the structural stability of liposomes. From these experimental results, the drug delivery carrier introduced with an amphiphilic polymer has an increased structural stability. In addition, it was confirmed that the bioactive ingredient, which is a macromolecule, can be easily delivered to the skin even in the emulsifier form used in cosmetics.

Claims (34)

A drug delivery carrier comprising a lipid structure or a polymer particle covalently bound to a cell penetrating peptide or a peptide chain comprising the peptide,
The lipid structure or polymer particle is a structure in which a physiologically active component is collected inside the structure,
The physiologically active ingredient is a water-soluble or water-insoluble macromolecule having a number average molecular weight or a weight average molecular weight of 500 Da or more,
The drug delivery carrier, wherein the cell-penetrating peptide is a peptide having a sequence of Gly (Arg) nGly Tyr Lys Cys (n = 1 to 20).   The drug delivery carrier according to claim 1, wherein the water-soluble or water-insoluble macromolecule has a number average molecular weight or a weight average molecular weight of 5,000 Da or more.   The drug delivery carrier according to claim 1 or 2, wherein the water-soluble or water-insoluble macromolecule has a number average molecular weight or a weight average molecular weight of 8,000 Da or more.   The said n is 3-9, The drug delivery carrier as described in any one of Claims 1-3 characterized by the above-mentioned.   The drug delivery carrier according to any one of claims 1 to 4, wherein the sequence is the sequence of SEQ ID NO: 1 (Gly Arg Arg Arg Arg Arg Arg Arg Arg Gly Tyr Lys Cys).   The drug delivery carrier according to any one of claims 1 to 5, wherein the lipid structure or polymer particle contains an amphiphilic polymer.   The drug delivery carrier according to any one of claims 1 to 6, wherein the cell-penetrating peptide or a peptide chain containing the cell-penetrating peptide is covalently bonded to an amphiphilic polymer. The amphiphilic polymer is an alkylated hyaluronic acid having an alkyl group attached to a side chain of hyaluronic acid, a poly (methacrylic acid-co-n-alkyl methacrylate) random copolymer represented by Formula 1. The drug delivery carrier according to claim 6 or 7, which is at least one selected from the group consisting of a random copolymer of poly (hydroxyethyl methacrylate-co-n-alkyl methacrylate) represented by Chemical Formula 2.




[In the above formula, n = 7-22,
x: y is 90: 10-50: 50 by molar ratio. ]   1 to 50% by weight of the amphiphilic polymer and 50 to 99% by weight of the lipid structure or polymer particles based on the total weight of the lipid structure or polymer particles and amphiphilic polymer. The drug delivery carrier according to any one of claims 6 to 8, which is contained in   The drug delivery carrier according to any one of claims 6 to 9, wherein the amphiphilic polymer has a number average molecular weight of 5,000 to 200,000 Da.   The drug delivery carrier according to any one of claims 1 to 10, wherein the drug delivery carrier comprises a lipid structure and further comprises a cholesterol derivative.   2. The lipid structure in the drug delivery carrier includes one or more of dioleyl phosphatidylethanolamine, phosphatidylcholine, and distearoylphosphatidylethanolamine-polyethylene glycol-maleimide (DSPE-PEG-Mal) complex. The drug delivery carrier according to any one of -11.   The drug delivery carrier is a dioleyl phosphatidylethanolamine: phosphatidylcholine: cholesterol derivative: distearoylphosphatidylethanolamine-polyethylene glycol-maleimide (DSPE-PEG-Mal) complex 1.0-2.0: 1.0-2. The drug delivery carrier according to any one of claims 1 to 12, which is contained in a molar ratio of 0.0: 1.0 to 3.0: 0.01 to 1.0.   The drug delivery carrier according to any one of claims 1 to 13, wherein the lipid structure is a liposome or an emulsion.   The drug delivery carrier according to claim 14, wherein the lipid component of the liposome or emulsion is a phospholipid having a fatty acid chain having 12 to 24 carbon atoms or a nitrogen lipid.   The phospholipids are yolk lecithin (phosphatidylcholine), soybean lecithin, lysolecithin, sphingomyelin, phosphatidic acid, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, diphosphatidylglycerol, lipiolipin cardio lip inlipin Natural phospholipids of plasmalogens, hydrogenated products that can be obtained from these phospholipids in the usual way, dicetyl phosphate, distearoyl phosphatidyl choline, distearoyl phosphatidyl-ethanolamine (DSPE), dioleoylphosphatidyl ethanol Amine, dipalmitoyl phosphatidylcholine, dipalmitoyl phosphatidylethanolamine, dipalmi It should be at least one selected from the group consisting of ylphosphatidylserine, eleostearoylphosphatidylcholine, eleostearoylphosphatidylethanolamine, synthetic lipids of eleostearoylphosphatidylserine, and fatty acid mixtures obtainable by hydrolysis thereof. The drug delivery carrier according to claim 15, wherein   The phospholipids are a combination of phosphatidylcholine and phosphatidylethanolamine, a combination of phosphatidylcholine and phosphatidylglycerol, a combination of phosphatidylcholine and phosphatidylinositol, a combination of phosphatidylcholine and phosphatidic acid, a combination of phosphatidylcholine and dioleoylphosphatidylethanolamine. Or a combination of phosphatidylcholine, dioleoylphosphatidylethanolamine, and phosphatidylserine.   The drug delivery carrier according to claim 17, wherein the combination ratio is such that the mixing ratio of the maximum component to the minimum component is 1: 5 or less.   The combination is phosphatidylcholine, dioleoylphosphatidylethanolamine and phosphatidylserine, and the combination ratio of the phosphatidylcholine: dioleoylphosphatidylethanolamine: phosphatidylserine combination is 1 to 4: 1 to 2: 1 to 2. The drug delivery carrier according to claim 17, wherein   The drug delivery carrier according to claim 15, wherein the lipid component is 0.001 to 20% by weight based on the total weight of the total liposome dispersion or emulsion.   21. The polymer particle according to any one of claims 1 to 20, wherein the polymer particle is compatible with a living body and is degraded in vivo without inducing inflammation or an immune reaction, and the degradation product thereof is also harmless in vivo. A drug delivery carrier according to 1.   The drug delivery carrier according to any one of claims 1 to 21, wherein the polymer particles are an amphiphilic polymer or a biodegradable aliphatic polyester polymer.   The drug delivery carrier according to any one of claims 1 to 22, wherein the polymer particle is a biodegradable aliphatic polyester polymer based on lactic acid and glycolic acid. The biodegradable aliphatic polyester polymer may be poly (D, L-lactic acid) (poly (D, L-lactic acid)), poly (L-lactic acid), poly (D -Lactic acid), poly (D, L-lactic acid-co-glycolic acid) represented by the following chemical formula 4 (poly (D, L-lactic acid-co-glycolic acid)), poly (D-lactic acid-co- Glycolic acid), poly (L-lactic acid-co-glycolic acid) poly (caprolactone), poly (valerolactone), poly (hydroxy butyrate) ), Poly (hydroxy valerate), poly (1,4-dioxane-2-one) (poly (1,4-dioxane-2-one)), poly (orthoester) (poly (Ortho ester)), and copolymers made from these monomers. The drug delivery carrier according to claim 22 or 23, wherein the drug delivery carrier is one or more selected from the group consisting of:


[In the above formula, n is an integer of 2 or more. ]


[Wherein, m and n are an integer of 2 or more, which may be the same or different from each other. ]   The drug delivery carrier according to any one of claims 22 to 24, wherein the biodegradable aliphatic polyester polymer has an average molecular weight of 500 Da to 100,000 Da.   The peptide (A) and the biodegradable aliphatic polyester polymer (B) are configured to be covalently bonded in the AB type or ABA type. A drug delivery carrier according to claim 1.   27. The covalent bond is formed by adding a base, a linker, or a multiligand compound between the peptide or a peptide chain containing a peptide and the lipid structure or polymer particle. Drug delivery carrier.   The drug delivery carrier according to any one of claims 1 to 27, wherein the drug delivery carrier has an average particle size of 1,000 nm or less.   A composition comprising the drug delivery carrier according to any one of claims 1 to 28 and a physiologically active ingredient collected inside the carrier.   The physiologically active ingredient is at least one selected from the group consisting of synthesized water-soluble macromolecular substances, macromolecular substances obtained by extraction of natural products, enzymes, EGF, proteins, peptides, and polysaccharides. Item 30. The composition according to item 29.   The composition according to claim 29 or 30, wherein the physiologically active ingredient collected in the drug delivery carrier is 0.01 to 30% by weight based on the total weight of the lipid structure or polymer particle.   The composition according to any one of claims 29 to 31, wherein the composition is a pharmaceutical composition having a dosage form selected from the group consisting of an external preparation for skin, an oral preparation and an injection.   The composition includes skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, essence, nutrition essence, pack, soap, 32. The cosmetic composition according to any one of claims 29 to 31, which is a cosmetic composition having any one or more dosage forms selected from the group consisting of cleansing foams, resin growths, cleansing creams, body lotions, and body cleansers. Composition. The bioactive component is a water-soluble macromolecule when the drug delivery carrier includes a lipid structure,
34. The composition according to any one of claims 29 to 33, wherein the drug delivery carrier is a water-insoluble macromolecule when it comprises polymeric particles.