KR20200005499A - Cosmetic composition for eliminating or adsorbing particulate matter comprising peptide complex as effective component - Google Patents

Abstract

The present invention relates to a cosmetic composition for removing or adsorbing fine dust, containing a fatty acid-amino acid complex or a fatty acid-oligopeptide complex as an active ingredient. The complex of the present invention has an excellent effect of removing fine dust without skin toxicity, and thus can be effectively used as a composition for preventing or treating various diseases caused by fine dust.

Description

Cosmetic composition for eliminating or adsorbing fine dust containing peptide complex as an active ingredient

The present invention relates to a cosmetic composition for removing or removing fine dust containing the peptide complex as an active ingredient.

Particulate matter (PM) is an air pollutant containing ionic components such as nitrates (NO ₃ ⁻ ), ammonium (NH ₄ ⁺ ) and sulfates (SO ₄ ^2- ), carbon compounds or metal compounds. It means a particulate matter having a diameter of 10 ㎛ or less floating in the long term. Fine dust is expressed as PM 10 when the particle diameter is 10 μm or less, and denoted as PM 2.5 when the particle diameter is less than 2.5 μm, and is referred to as ultrafine dust or ultrafine dust. Most of the ultra fine dust is caused by anthropogenic factors such as automobile exhaust, industrial process, and dust blowing from roads.

When exposed to fine dust for a long time, the immunity is sharply lowered and can be exposed to various diseases such as cardiovascular disease, skin disease and eye disease as well as respiratory diseases such as cold, asthma and bronchitis. Fine dust has been reported to exacerbate respiratory diseases such as asthma and reduce lung function.In particular, ultrafine dust particles are so fine that they do not filter through the nasal mucosa and immediately penetrate into the alveoli, leading to the prevalence and prevalence of lung disease. Increase mortality. In addition, fine dust can enter the body not only through the nose, mouth, but also through the skin. Fine dust is up to 20 times smaller than the size of the pores, so it is difficult to penetrate into the pores and are difficult to remove.Increasing inflammatory cytokines and free radicals by the absorbed fine dust can reduce skin immunity and cause acne. It further accelerates skin aging such as wrinkles, skin dryness and pigmentation. Specifically, fine dust may cause inflammatory reactions to damage the skin barrier and exacerbate atopic dermatitis, and produce reactive oxygen species in mitochondria to reduce collagen synthesis and increase collagen breakdown to reduce skin aging. May cause In addition, polynuclear aromatic hydrocarbons (PAH) adhered to fine dust can increase melanocytes and increase pigment spots on the face.In particular, Asians have darker skin than white, which increases pigment spots due to fine dust. It is very likely to cause dermatitis, burning and pruritus in people with sensitive skin. Long-term exposure to fine dust can cause irreversible skin damage that can cause serious illnesses such as skin cancer, as the skin loses its ability to protect itself from environmental stressors.

Recently, as the danger and damage of fine dust has emerged seriously, the interest in the development of products for the prevention of fine dust has increased, and the sales volume of cosmetics, health functional foods, air cleaners, and masks related to fine dust has increased rapidly. There is a continuing need for the development of a cosmetic composition capable of removing fine dust or significantly preventing the adsorption of fine dust. Peptide materials are representative of high functional bio raw materials that meet this trend. Depending on the arrangement and structure of the amino acid sequence in the peptide has a biological activity in vivo or in the skin can be applied to a variety of biological materials development, semi-treatment for the improvement of wrinkles, whitening, trouble skin and various skin diseases It can also be applied to the development of sexual products, which is expected to have an excellent industrial utility value as a cosmetic and pharmaceutical material.

Meanwhile, Korean Patent No. 173941 discloses a 'cosmetic composition for adsorption and removal of fine dust as an active ingredient of coral lake and Bengal rubber resin extract', and Korean Patent No. 1850313 uses' Chrysanthemum extract as an active ingredient. There is disclosed a cosmetic composition for removing heavy metals or fine dust containing the fine dust removal or adsorption cosmetic composition containing the peptide complex of the present invention as an active ingredient.

The present invention has been derived by such a demand, and the present inventors have described amino acids; Or a fatty acid-amino acid complex and a fatty acid-oligopeptide complex by combining saturated fatty acid or unsaturated fatty acid to the N-terminal, C-terminal or side chain of the oligopeptide, and the complex reduces the cytotoxicity caused by the fine dust and The present invention was completed by confirming that coagulation and fine dust could be removed.

In order to solve the above problems, the present invention is an amino acid; Or N terminal, C terminal or side chain of the oligopeptide; provides a cosmetic composition for removing fine dust or adsorption containing a complex of fatty acids bonded as an active ingredient.

Since the peptide complex according to the present invention is excellent in the effect of removing the fine dust by aggregation or binding to the fine dust as well as cell protective effect against fine dust, the composition comprising the peptide complex of the present invention is a variety of It is expected to be usefully used as a cosmetic or pharmaceutical composition for the prevention and treatment of diseases.

1 is a conceptual diagram illustrating a method of binding a fatty acid to an N-terminus (A), C-terminus (B) or side chain (C) of an oligopeptide (pentapeptide).
2 to 4 show the results of analyzing the peptide complex (Table 5-Division 9) purified through High Performance Liquid Chromatography (HPLC).
5 to 15 are the results of confirming the cell viability after treating HaCaT cells with ultrafine dust (diesel exhaust particles; fine dust with a diameter of 2.5 μm or less, PM2.5) and the peptide complex of the present invention at the same time. NC (negative control): 0.2% DMSO (dimethyl sulfoxide) / Dulbecco's Modified Eagle Medium (DMEM) treatment, PC (positive control): 10% DMSO / DMEM treatment.
Figure 16 is a graph showing the cytotoxicity by treating the solution containing ultra-fine dust in human lung cancer epithelial cell line (A549) by concentration.
17 is a result of confirming cell viability after simultaneously processing a complex of ultrafine dust and saturated fatty acid bound to the oligopeptide in A549 cells. The information of the graph X-axis sample is the same as in Table 10 below.
FIG. 18 shows the aggregation effect of the peptide complex (complex of capric acid (C10) bonded to the 9th K side chain of the oligopeptide (llwialrkk) of SEQ ID NO: 30) and ultrafine dust using silver staining; Is a SDS-PAGE gel picture analyzed.
19 is a graph showing particle removal efficiencies of commercially available masks (HIP's) and fine dust masks (KF84) using a dust collection (particle removal) efficiency evaluation system.
20 and 21 illustrate a general mask sprayed with nothing using a dust collection efficiency evaluation system, a general mask sprayed with purified water, and a normal mask sprayed with a 250 ppm solution of a fatty acid-oligopeptide complex (distilled water dilution). It is a graph showing the ultrafine particle removal efficiency.

In order to achieve the object of the present invention, the present invention is amino acid; Or N terminal, C terminal or side chain of the oligopeptide; provides a cosmetic composition for removing fine dust or adsorption containing a complex of fatty acids bonded as an active ingredient.

In the present invention, the fine dust is a concept including the ultra fine dust, the term 'fine dust' refers to dust having a diameter of 10 ㎛ or less, the term 'ultra fine dust' refers to dust having a diameter of 2.5 ㎛ or less It may include, but is not limited to, air pollutants such as yellow dust, automobile exhaust gas, factory smoke, domestic combustion gas, and heavy metals.

In the present invention, the term 'fine dust adsorption' may mean to remove by adsorbing fine substances remaining in the pores in the skin. Treatment of cosmetic compositions based on fatty acid and amino acid complexes or fatty acid and oligopeptide complexes improves adsorption to fine dust in the skin and is hypoallergenic to the skin and eliminates skin irritation or skin problems that may occur when fine dust remains. Can be.

The term 'N-terminal and C-terminal' in the present invention means one end having an amino group (-NH ₂ ) and the other end having a carboxyl group (-COOH) in the amino acid sequence of the peptide, the term 'side chain' It refers to an aliphatic carbon chain attached to a ring in a carbon chain or a cyclic compound in which branches are divided from straight chain carbon atoms in which carbon atoms are regularly connected.

In the cosmetic composition for removing fine dust or adsorption of the present invention, the amino acid is alanine (A), cysteine (C), aspartic acid (D), glutamic acid (G), phenylalanine (Phenylalanine , F), Glycine (G), Histidine (H), Isoleucine (I), Lysine (Lysine, K), Leucine (L), Methionine (M), Asparagine , N), Pyrrolysine (O), Proline (P), Glutamine (Q), Arginine (Arginine, R), Serine (S), Threonine (Threonine, T), Seleno At least one selected from the group consisting of cysteine (U), valine (V), tryptophan (W) and tyrosine (Y), preferably glycine (G), glutamic acid (E) or Lysine (K), but is not limited thereto. The amino acid may include both D-type and L-type amino acids, but is not limited thereto.

In addition, in the cosmetic composition for removing fine dust or adsorption of the present invention, the oligopeptide is a peptide consisting of two or more amino acids, preferably AAPV (SEQ ID NO: 1), AAP (SEQ ID NO: 2), AA (SEQ ID NO: 3) ), GHK (SEQ ID NO: 4), GH (SEQ ID NO: 5), KTTKS (SEQ ID NO: 6), KTTK (SEQ ID NO: 7), KTT (SEQ ID NO: 8), KT (SEQ ID NO: 9), EEMQRR (SEQ ID NO: 10) , EEMQR (SEQ ID NO: 11), EEMQ (SEQ ID NO: 12), EEM (SEQ ID NO: 13), EE (SEQ ID NO: 14), KKKKKKKKK (SEQ ID NO: 15), KKKKK (SEQ ID NO: 16), KKK (SEQ ID NO: 17), KK (SEQ ID NO: 18), GPO (SEQ ID NO: 19), GP (SEQ ID NO: 20), GQPR (SEQ ID NO: 21), GQP (SEQ ID NO: 22), GQ (SEQ ID NO: 23), TTKS (SEQ ID NO: 24), TKS (SEQ ID NO: 25), TK (SEQ ID NO: 26), RRRRRRRRR (SEQ ID NO: 27), RRRRR (SEQ ID NO: 28), RRR (SEQ ID NO: 29), and LLWIALRKK (SEQ ID NO: 30), This is not restrictive. The amino acid of the oligopeptide may be composed of D-type amino acid or L-type amino acid, and the amino acid PO of SEQ ID NO: 19 may be O-hydroxyproline, but is not limited thereto.

In the cosmetic composition for removing or removing fine dust of the present invention, the fatty acid may be saturated or unsaturated fatty acid, but is not limited thereto.

The saturated fatty acid according to the present invention is caproic acid (Caproic acid, C6: 0), caprylic acid (Caprylic acid, C8: 0), Pelargonic acid (Pelargonic acid, C9: 0), capric acid (Capric acid, C10: 0), Undecylic acid (C11: 0), Lauric acid (C12: 0), Myristic acid (C14: 0), Pentadecylic acid (C15: 0 ), Palmitic acid (C16: 0), Margaric acid (C17: 0), Stearic acid (Ctearic acid, C18: 0), Nonadecylic acid (C19: 0), Sub Arachidic acid (C20: 0), Henicosylic acid (C21: 0), Behenic acid (C22: 0), Tricosylic acid (Cricosylic acid. C23: 0), Lignose Lignoceric acid (C24: 0), pentacosylic acid (C25: 0), cerotic acid (C26: 0), heptacosylic acid (C27: 0), montanic acid , C28: 0), nonacosylic acid (C29: 0), melissic acid (C30: 0), henatriacontylic acid (C31: 0), lacse Acid (Lacceroic acid, C32: 0), Psyllic acid (C33: 0), Gedic acid (C34: 0), Ceroplastic acid (C35: 0), and hexatriacontylic acid ( Hexatriacontylic acid, C36: 0), preferably at least one selected from the group consisting of caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, margaric acid, stearic acid, behenic acid or It may be lignoseric acid, but is not limited thereto.

In addition, the unsaturated fatty acid may be palmitoleic acid (Palmitoleic acid, C16: 1), oleic acid (Oleic acid, C18: 1), elideic acid (Elaidic acid, C18: 1), petroleic acid (Petroselinic acid, C18: 1) ), Vaccenic acid (C18: 1), Gondoic acid (C20: 1), Erucic acid (C22: 1), Nervonic acid (C24: 1), Linoleic acid acid, C18: 2), cis-Linolenic acid (C18: 3), Punicic acid (C18: 3), Eleostearic acid (C18: 3), Stearic acid ( stearidonic acid (C18: 4), Arachidonic acid (C20: 4), Eicosapentaenoic acid (C20: 5), Docosapentaenoic acid (C22: 5), Adrenic acid ( Adrenic acid (C22: 4), docosahexaenoic acid (Docosahexaenoic acid (C22: 6) and ricinoleic acid (C18: 1) may be one or more selected from the group consisting of, preferably oleic acid, linoleic acid, Palmitoleic acid, elideic acid, petroleum acid, Greater sensan, Oregon bankruptcy, erucic acid, Nerja acid, cis-on-linolenic acid, fumaric acid Dubrovnik, eleostearic acid, but acid Toko four Penta yen or receiver linoleate sanil, but not limited thereto.

The fine dust removal or adsorption cosmetic composition of the present invention is an amino acid; Or an N-terminal, C-terminal or side chain of the oligopeptide; a complex of fatty acids and amino acids bound to a fatty acid or a complex of fatty acids and oligopeptides, wherein the complex of fatty acids and amino acids is glycine (G), glutamic acid (E) or lysine ( K) may be an unsaturated fatty acid oleic acid is bound, but is not limited thereto, the complex of the fatty acid and oligopeptide is SEQ ID NO: 1, 4, 6, 7, 8, 10, 11, 15, 16, 17, 19 Caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, mar, at the N-terminal, C-terminal, or side chain of any of the oligopeptides consisting of the amino acid sequences 21, 27, 28, 29, and 30 At least one saturated fatty acid selected from the group consisting of garic acid, stearic acid, behenic acid and lignoseric acid; Or oleic acid, linoleic acid, palmitoleic acid, elideic acid, petroleic acid, barksenic acid, gondonic acid, erucic acid, nerbonic acid, cis-linolenic acid, punic acid, eleostearic acid, docosapentaenoic acid, and rishi One or more unsaturated fatty acids selected from the group consisting of nooleic acid; may be combined, but is not limited thereto.

In the cosmetic composition for removing fine dust or adsorption of the present invention, the complex in which a fatty acid is bound to the side chain of the oligopeptide is preferably an oligopeptide (LLWIALRKK, L-type amino acid; llwialrkk, D-type amino acid) amino acid of SEQ ID NO. Lysine 9 in the sequence (Lysine, K); Fourth lysine in the oligopeptide amino acid sequence of SEQ ID NO: 6 (KTTKS) and SEQ ID NO: 7 (KTTK); Fatty acid may be bound to the fifth lysine in the oligopeptide amino acid sequence of SEQ ID NO: 16 (KKKKK), but is not limited thereto.

In addition, the peptide complex of the present invention may be in the form of an amino group (-NH ₂ ) or a carboxyl group (-COOH) is bonded to the N- or C- terminal, specifically, NH ₂ -amino acid-fatty acid-COOH; NH ₂ -oligopeptide-fatty acid-COOH; COOH-amino acid-fatty acid-NH ₂ ; COOH-oligopeptide-fatty acid-NH ₂ ; NH ₂ -fatty acid-amino acid-COOH; NH ₂ -fatty acid-oligopeptide-COOH; COOH-fatty acid-amino acid-NH ₂ ; And COOH-fatty acid-oligopeptide-NH ₂ form, but is not limited thereto.

Amino acids of the invention; Or the N-terminus, C-terminus or side chain of the oligopeptide; the conjugated saturated fatty acid or unsaturated fatty acid is not toxic to normal skin cells, can reduce the cytotoxicity caused by fine dust, ultrafine particles It has a removal (dust collection) effect.

Specifically, the capric acid of the present invention is conjugated to the oligopeptide of SEQ ID NO: 6, 7, 15, 16, 27, 28 or 29; A complex in which palmitic acid and an oligopeptide of SEQ ID NO: 4, 6, 7, 15, 16, 27, 28, 29, or 30 are bound; Complexes in which oleic acid and oligopeptides of SEQ ID NOs: 1, 4, 6, 7, 8, 11, 15, 16, 21, 27, 28 or 30 are bound; A complex in which linoleic acid and an oligopeptide of SEQ ID NO: 10, 16, 27, 28 or 30 are bound; A complex in which caprylic acid and an oligopeptide of SEQ ID NO: 15, 16, 27 or 28 are bound; A complex in which lauric acid and an oligopeptide of SEQ ID NO: 4, 15, 16, 17, 27 or 28 are bound; A complex in which myristic acid and an oligopeptide of SEQ ID NO: 10, 15, 16, 27 or 28 are bound; A complex in which margaric acid and an oligopeptide of SEQ ID NO: 15, 16, 27, 28 or 30 are bound; A combination of stearic acid and an oligopeptide of SEQ ID NO: 15, 16, 27, 28 or 30; A complex in which behenic acid and an oligopeptide of SEQ ID NO: 15, 16, 19, 27, 28 or 30 are bound; A complex in which lignoseric acid and an oligopeptide of SEQ ID NO: 15, 16, 27, 28 or 30 are bound; A complex in which palmitoleic acid and an oligopeptide of SEQ ID NO: 1, 15. 21 or 27 are bound; A complex in which elideic acid and an oligopeptide of SEQ ID NO: 6, 15, 16, 19 or 27 are bound; Complexes in which petroleic acid and an oligopeptide of SEQ ID NO: 6, 15, 16, 27 or 30 are bound; A complex in which a vexenic acid and an oligopeptide of SEQ ID NO: 15, 27 or 28 are bound; A complex in which a gondonic acid and an oligopeptide of SEQ ID NO: 6, 15, 27, 28 or 30 are bound; A complex in which erucic acid and an oligopeptide of SEQ ID NO: 15, 27, 28 or 30 are bound; A complex in which nerbonic acid and an oligopeptide of SEQ ID NO: 4, 6, 15, 16, 27, 28 or 30 are bound; A complex of cis-linolenic acid bound to an oligopeptide of SEQ ID NO: 15, 16, 27, 28 or 30; A complex in which a funic acid and an oligopeptide of SEQ ID NO: 6, 15, 16 or 27 are bound; A complex in which eleostearic acid and an oligopeptide of SEQ ID NO: 6, 15, 27 or 30 are bound; A complex in which docosapentaenoic acid and an oligopeptide of SEQ ID NO: 1, 15, 16 or 29 are bound; Or a complex in which ricinoleic acid and an oligopeptide of SEQ ID NO: 15, 27 or 30 are combined; is excellent in reducing the cytotoxicity increased by ultrafine particles and increasing the ultrafine particle removal (dust collection) rate.

Cosmetic composition for removing or adsorbing fine dust of the present invention is selected from solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, foundations, wax foundations and sprays. It is preferably one of the formulations, and more preferably, external skin ointments, creams, supple cosmetics, nourishing cosmetics, packs, essences, hair tonics, shampoos, rinses, hair conditioners, hair treatments, gels, skin lotions, skin softeners , Skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, eye cream, moisture cream, hand cream, foundation, nutrition essence, sunscreen, soap, cleansing foam, cleansing lotion, cleansing cream It may have any one formulation selected from the group consisting of, body lotion and body cleanser, but is not limited thereto . The composition of each of these formulations may contain a variety of bases and additives necessary for the formulation of the formulation and are suitable, and the types and amounts of these components can be readily selected by those skilled in the art.

When the formulation of the present invention is a paste, cream or gel, animal carriers, plant fibers, waxes, paraffins, starches, tracantes, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc acid may be used as carrier components. Can be.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the dosage form of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable fats and oils, linolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Example  One. Peptide  Preparation of the complex

Amino acid raw materials used in the synthesis of the complex were purchased from GLS (GL Biochem, Shanghai), and fatty acids were purchased from TCI chemicals (India) and Sigma (Sigma Aldrich, US) (Table 1), dimethylformamide (dimethylformamide, DMF), diisopropylethylamine (N, N-diisopropylethylamine, DIEA), dichloromethane (DCM) and piperidine were purchased from Daejungchem, Korea.

Fatty Acid Purchase Information division fatty acid Where to buy One Capric acid TCI (TCI chemicals, Korea), D0017 2 Palmitic acid TCI (TCI chemicals, Korea), P0002 3 Arachnic acid TCI (TCI chemicals, Korea), E0006 4 Oleic acid TCI (TCI chemicals, Korea), O0011 5 Linoleic acid Sigma (Sigma Aldrich, Korea), L1376 6 Eicosapentaenoic acid TCI (TCI chemicals, Korea), E0441 7 Caprylic acid TCI (TCI chemicals, Korea), E0120 8 Lauric acid TCI (TCI chemicals, Korea), A0932 9 Myristic acid TCI (TCI chemicals, Korea), M0477 10 Margaric acid TCI (TCI chemicals, Korea), C0950 11 Stearic acid TCI (TCI chemicals, Korea), S0235 12 Behenic acid TCI (TCI chemicals, Korea), B1248 13 Lignoseric acid TCI (TCI chemicals, Korea), L0111 14 Palmitoleic acid TCI (TCI chemicals, Korea), H0501 15 Elide acid TCI (TCI chemicals, Korea), O0010 16 Petroselin acid Sigma (Sigma Aldrich, Korea), P8750 17 Bachsensan TCI (TCI chemicals, Korea), M2308 18 Kundosan Sigma (Sigma Aldrich, Korea), E3635 19 Erucic acid TCI (TCI chemicals, Korea), D0965 20 Nerbonic acid TCI (TCI chemicals, Korea), T1642 21 Funic acid Cayman (Cayman chemicals, USA), 22976EL 22 Eleostearic acid Chemical Book USA, 10008349 23 Arachidonic acid TCI (TCI chemicals, Korea), A0781 24 Docosapentaenoic Acid TCI (TCI chemicals, Korea), D2964 25 Cis-linolenic acid TCI (TCI chemicals, Korea), L0152 26 Docosahexaenoic acid TCI (TCI chemicals, Korea), D2226 27 Ricinoleic acid TCI (TCI chemicals, Korea), R0027

1-1. Resin Swelling

A solid-phase synthesis reactor equipped with a filtration membrane was used, and peptide synthesis having a carboxyl group (-COOH) at the end of the complex was performed using 2-chlorotritylchloride resin (Bead Tech, Korea). , Peptide synthesis terminated with a peptide bond (-CONH ₂ ) at the end of the complex using a link amide resin (GLS, China). The resin was prepared by swelling the resin for 20 minutes using dichloromethane and dimethylformamide.

1-2. Amino acid loading

Synthesis with chlorotrityl chloride resin involves loading the first amino acid into the resin. The resin was removed from the solvent through a filtration membrane under reduced pressure. 3 to 5 equivalents of amino acid in the resin was completely dissolved in DMF, and then added to chlorotrityl chloride resin, and an amount corresponding to 3 to 5 equivalents of chlorotrityl chloride resin was added to diisopropylethylamine in consideration of density. Thereafter, the reactor was reacted at a temperature of 24 ° C. to 32 ° C. for at least 5 hours.

1-3. Resin Fmoc Deprotection

Synthesis using the chlorotrityl chloride resin or linkamide resin included a process of deprotection of Fluorenylmethyloxycarbonyl chloride (Fmoc). In the resin Fmoc deprotection process, the solvent was removed through a filtration membrane under reduced pressure, washed for 5 minutes with DMF added with 20% (v / v) piperidine, and then again for 10 minutes. The reaction solution was removed by filtration under reduced pressure, and washed six more times for 5 minutes using DCM or DMF.

1-4. Fatty acid-amino acid complexes and fatty acid- Oligopeptide  Complex synthesis

3 to 5 equivalents of amino acid in the link amide resin was completely dissolved in DMF, and the solvent was removed to the link amide resin. As a coupling reagent, 2 M HOBt / DIC (hydroxybenzotriazole / diisopropylcarbodiimide) was added to the amino acid equivalent and the link amide resin amount. Then, the synthesis was carried out at 24 ~ 32 ℃ 5 hours or more using a reactor. After the reaction was completed, the solvent was removed through a filtration membrane under reduced pressure, and then washed six times with clean DMF five times. After washing, amino acids were coupled in sequence in the same manner as the amino acid binding method. Thereafter, 3 equivalents of fatty acid and 2 M HOBt / DIC were added to the amino acid equivalent and the resin amount in the resin synthesized peptide. After using the reactor for 5 hours or more, it was reacted at 24 ~ 32 ℃ temperature.

1-5. Cleavage

After the reaction was completed, the solvent was removed through the filter membrane under reduced pressure, and then washed twice with clean DMF twice for 2 minutes and twice with DCM for 2 minutes, and then the solvent was almost removed through the filter membrane under reduced pressure. The dried peptide complex resin was separated for 4 hours using 70% (v / v) TFA / 29% (v / v) DCM / 1% (v / v) H ₂ O solution.

1-6. Crystallize

The separated solvent was extracted by recrystallization of the crude product using ethyl ether. Specific synthesis process is shown in Table 2 below.

Peptide complex synthesis process of the present invention * Resin substitution rate:
Rink amide MBHA resin (substitution rate: 0.54 mmole / g)
Chlorotrityl chloride resin (substitution rate: 1.25 mmol / g)
* Amino acid: 3 ~ 10 eq
Coupling reagents: DIC, HOBt, DMF
* Coupling time: 6 hours
Coupling Temperature: 24 ~ 32 ℃
* Separation: 70% TFA / 29% DCM / 1% H2O, 4 hours, or 5% TFA / 95% DCM 5 minutes
* Extraction: Ethyl ether

1-7. Synthetic products

Solid phase peptide synthesis (SPPS) through 6 to 24 carbon atoms of fatty acids (Table 3) and the amino acids glycine (G), glutamic acid (E), lysine (K) or oligos of SEQ ID NOs: 1 to 30 New complexes with peptides have been developed. Specifically, amino acids; Or an amino group (NH ₂ ) present at the N-terminus of the oligopeptide and a carboxyl group (COOH) of a fatty acid (FIG. 1A), and an amino acid using ethylene glycol as a linker; Alternatively, the carboxyl group of the fatty acid present at the C terminus of the oligopeptide may bind the carboxyl group of the fatty acid (FIG. 1B), and the carboxyl group of the fatty acid may be bonded to the side chain present in the lysine (K) of the oligopeptide (FIG. 1C).

 Finally, the synthesized complexes of amino acids or oligopeptides and fatty acids are shown in Tables 4 and 10, in which the uppercase letters indicate L-type amino acids and the lowercase letters indicate D-type amino acids. amino acid; Alternatively, the complexes in which the fatty acids are bound to the N-terminus of the oligopeptides are described in the order of fatty acids-peptide, and the complexes in which the fatty acids are bound to the C-terminus of the amino acids or oligopeptides are described in the order of peptide-fatty acids, in the side chains of the oligopeptides. Complex conjugated fatty acid is described by underlined (_) to the amino acid attached to the side chain to which the fatty acid is bound and then indicated in parentheses in the fatty acid.

Fatty acid types [Saturated Fatty Acids] Systematic name Common name Formula Symbol
( Carbon number
: Double bond water ) Caproic acid Hexanoic acid CH ₃ (CH ₂ ) ₄ COOH C6: 0 Caprylic acid Octanoic acid CH ₃ (CH ₂ ) ₆ COOH C8: 0 Capric acid Decanoic acid CH ₃ (CH ₂ ) ₈ COOH C10: 0 Lauric acid Dodecanoic acid CH ₃ (CH ₂ ) ₁₀ COOH C12: 0 Myristic acid Tetradecanoic acid CH ₃ (CH ₂ ) ₁₂ COOH C14: 0 Palmitic acid Hexadecanoic acid CH ₃ (CH ₂ ) ₁₄ COOH C16: 0 Margaric acid Heptadecanoic acid CH ₃ (CH ₂ ) ₁₅ COOH C17: 0 Stearic acid Octadecanoic acid CH ₃ (CH ₂ ) ₁₆ COOH C18: 0 Arachidic acid Eicosanoic acid CH ₃ (CH ₂ ) ₁₈ COOH C20: 0 Behenic acid Docosanoic acid CH ₃ (CH ₂ ) ₂₀ COOH C22: 0 Lignoceric acid Tetracosanoic acid CH ₃ (CH ₂ ) ₂₂ COOH C24: 0 [Unsaturated fatty acids] Systematic name Common name Formula Symbol Palmitoleic acid (Z) -9-hexadecenoic acid C ₁₆ H ₃₀ O ₂ C16: 1 Oleic acid (Z) -9-octadecenoic acid C ₁₈ H ₃₄ O ₂ C18: 1 Elaidic acid (E) -9-octadecenoic acid C ₁₈ H ₃₄ O ₂ C18: 1 Petroselinic acid (Z) -6-octadecaenoic acid C ₁₈ H ₃₄ O ₂ C18: 1 Vaccenic acid (E) -11-octadecenoic acid C ₁₈ H ₃₄ O ₂ C18: 1 Gondoic acid (Z) -11-eicosenoic acid C ₂₀ H ₃₈ O ₂ C20: 1 Erucic acid (Z) -13-docosenoic acid C ₂₂ H ₄₂ O ₂ C22: 1 Nervonic acid (Z) -15-tetracosenoic acid C ₂₄ H ₄₆ O ₂ C24: 1 Linoleic acid (LA) (Z, Z) -9,12-octadecadienoic acid C ₁₈ H ₃₂ O ₂ C18: 2 cis-linolenic acid (CLA) (E, E, E) -9,12,15-octadecatrienoic acid C ₁₈ H ₃₀ O ₂ C18: 3 Punicic acid (Z, E, Z) -9,11,13-octadecatrienoic acid C ₁₈ H ₃₀ O ₂ C18: 3 Oleostearynic acid (E, E, E) -9,11,13-octadecatrienoic acid C ₁₈ H ₃₀ O ₂ C18: 3 Arachidonic acid (AA) ( all -Z) -5,8,11,14-eicosatetraenoic acid C ₂₀ H ₃₂ O ₂ C20: 4 Eicosapentaenoic acid (EPA),
Timnodonic acid (all-Z) -5,8,11,14,17-icosapentaenoic acid C ₂₀ H ₃₀ O ₂ C20: 5 Docosapentaenoic acid (DPA), Clupanodonic acid ( all -Z) -4,8,12,15,19-docosapentaenoic acid C ₂₂ H ₃₄ O ₂ C22: 5 Docosahexaenoic acid (DHA) ( all -Z) -4,7,10,13,16,19-docosahexaenoic acid C ₂₂ H ₃₂ O ₂ C22: 6 Ricinoleic acid (R) -12-hydroxy- (Z) -9-octadecenoic acid C ₁₈ H ₃₄ O ₂ C18: 1 Omega 3 Fatty Acid Alpha-linolenic acid (ALA), Eicosapentaenoic acid (EPA), Docosahexaenoic acid (DHA) Omega 6 Fatty Acid Linoleic acid (LA), Gamma-linolenic acid (GLA), Arachidonic acid (AA) Omega 9 Fatty Acid Oleic acid

Types of synthesized fatty acid-amino acid complexes and fatty acid-oligopeptide complexes division name Carbon number division name Carbon number




One

Caprylic acid-AAPV C8





4







Caprylic acid-EEMQRR C8 Capric acid-AAPV C10 Capric acid-EEMQRR C10 Lauric acid-AAPV C12 Lauric acid-EEMQRR C12 Myristic acid-AAPV C14 Myristic acid-EEMQRR C14 Palmitoleic acid-AAPV C16 Palmitoleic acid-EEMQRR C16 Margaric acid-AAPV C17 Margaric acid-EEMQRR C17 Oleic acid-AAPV C18 Oleic acid-EEMQRR C18 Arachidic acid-AAPV C20 Arachidic acid-EEMQRR C20 Behenic acid-AAPV C22 Behenic acid-EEMQRR C22 Lignoceric acid-AAPV C24 Lignoceric acid-EEMQRR C24 Oleic acid-AAP C18 Oleic acid-EEMQR C18 Oleic acid-AA C18 Oleic acid-EEMQ C18




2





Caprylic acid-GHK C8 Oleic acid-EEM C18 Capric acid-ghk C10 Oleic acid-EE C18 Lauric acid-ghk C12 Oleic acid-E C18 Myristic acid-GHK C14




5




Caprylic acid-KKK C8 Palmitoleic acid-ghk C16 Capric acid-KKK C10 Margaric acid-GHK C17 Lauric acid-KKK C12 Oleic acid-GHK C18 Myristic acid-KKK C14 Arachidic acid-GHK C20 Palmitoleic acid-KKK C16 Behenic acid-GHK C22 Margaric acid-KKK C17 Lignoceric acid-GHK C24 Oleic acid-KKK C18 Oleic acid-GH C18 Arachidic acid-KKK C20 Oleic acid-g C18 Behenic acid-KKK C22




3





Caprylic acid-KTTKS C8 Lignoceric acid-KKK C24 Capric acid-KTTKS C10 Oleic acid-KK C18 Lauric acid-KTTKS C12 Oleic acid-K C18 Myristic acid-KTTKS C14




6




Caprylic acid-GPO (O-hydroxyproline) C8 Palmitoleic acid-KTTKS C16 Capric acid-GPO (O-hydroxyproline) C10 Margaric acid-KTTKS C17 Lauric acid-GPO (O-hydroxyproline) C12 Oleic acid-KTTKS C18 Myristic acid-GPO (O-hydroxyproline) C14 Arachidic acid-KTTKS C20 Palmitoleic acid-GPO (O-hydroxyproline) C16 Behenic acid-KTTKS C22 Margaric acid-GPO (O-hydroxyproline) C17 Lignoceric acid-KTTKS C24 Oleic acid-GPO (O-hydroxyproline) C18 Oleic acid-KTTK C18 Arachidic acid-GPO (O-hydroxyproline) C20 Oleic acid-KTT C18 Behenic acid-GPO (O-hydroxyproline) C22
Lignoceric acid-GPO (O-hydroxyproline) C24 Oleic acid-GP C18

Type of synthesized fatty acid-oligopeptide complex division name Carbon number division name Carbon number




7





Caprylic acid-GQPR C8












9
















Palmitic acid-AAPV C16: 0 Capric acid-GQPR C10 Palmitic acid-AAP C16: 0 Lauric acid-GQPR C12 Palmitic acid-AA C16: 0 Myristic acid-GQPR C14 Palmitic acid-GHK C16: 0 Palmitoleic acid-GQPR C16 Palmitic acid-gh C16: 0 Margaric acid-GQPR C17 Palmitic acid-KTTKS C16: 0 Oleic acid-GQPR C18 Palmitic acid-KTTK C16: 0 Arachidic acid-GQPR C20 Palmitic acid-KTT C16: 0 Behenic acid-GQPR C22 Palmitic acid-KT C16: 0 Lignoceric acid-GQPR C24 Palmitic acid-EEMQRR C16: 0 Oleic acid-GQP C18 Palmitic acid-EEMQR C16: 0 Oleic acid-gq C18 Palmitic acid-EEMQ C16: 0









8










Capric acid-AAP C10: 0 Palmitic acid-EEM C16: 0 Capric acid-AA C10: 0 Palmitic acid-EE C16: 0 Capric acid-gh C10: 0 Palmitic acid-KKK C16: 0 Capric acid-KTTK C10: 0 Palmitic acid-KK C16: 0 Capric acid-KTT C10: 0 Palmitic acid-gp C16: 0 Capric acid-KT C10: 0 Palmitic acid-GQPR C16: 0 Capric acid-EEMQR C10: 0 Palmitic acid-GQP C16: 0 Capric acid-EEMQ C10: 0 Palmitic acid-gq C16: 0 Capric acid-EEM C10: 0 Palmitic acid-LLWIALRKK C16: 0 Capric acid-ee C10: 0 Palmitic acid-GPO C16: 0 Capric acid-KK C10: 0 Palmitic acid-llwialrkk C16: 0 Capric acid-gp C10: 0 Palmitic acid-TK C16: 0 Capric acid-GQP C10: 0 Palmitic acid-TTKS C16: 0 Capric acid-gq C10: 0 Palmitic acid-TKS C16: 0 Capric acid-TK C10: 0 Palmitic acid-KKKKK C16: 0 Capric acid-TTKS C10: 0 Palmitic acid-KKKKKKKKK C16: 0 Capric acid-TKS C10: 0 Palmitic acid-RRR C16: 0 Capric acid-KKKKK C10: 0 Palmitic acid-RRRRR C16: 0 Capric acid-KKKKKKKKK C10: 0 Palmitic acid-RRRRRRRRR C16: 0 Capric acid-RRR C10: 0

Capric acid-RRRRR C10: 0 Capric acid-RRRRRRRRR C10: 0

Type of synthesized fatty acid-oligopeptide complex division name Carbon number division name Carbon number




10




Oleic acid-KT C18: 1

12

Caprylic acid-RRR C8: 0 Oleic acid-LLWIALRKK C18: 1 Caprylic acid-RRRRR C8: 0 Oleic acid-llwialrkk C18: 1 Caprylic acid-RRRRRRRRR C8: 0 Oleic acid-TK C18: 1 Caprylic acid-KKKKK C8: 0 Oleic acid-TTKS C18: 1 Caprylic acid-KKKKKKKKK C8: 0 Oleic acid-TKS C18: 1

13

Lauric acid-RRR C12: 0 Oleic acid-KKKKK C18: 1 Lauric acid-RRRRR C12: 0 Oleic acid-KKKKKKKKK C18: 1 Lauric acid-RRRRRRRRR C12: 0 Oleic acid-RRR C18: 1 Lauric acid-KKKKK C12: 0 Oleic acid-RRRRRR C18: 1 Lauric acid-KKKKKKKKK C12: 0 Oleic acid-RRRRRRRRR C18: 1

14

Myristic acid-RRR C14: 0













11















Linoleic acid-AAPV C18: 2 Myristic acid-RRRRR C14: 0 Linoleic acid-aap C18: 2 Myristic acid-RRRRRRRRR C14: 0 Linoleic acid-aa C18: 2 Myristic acid-KKKKK C14: 0 Linoleic acid-ghk C18: 2 Myristic acid-KKKKKKKKK C14: 0 Linoleic acid-gh C18: 2


15


Margaric acid-LLWIALRKK C17: 0 Linoleic acid-KTTKS C18: 2 Margaric acid-llwialrkk C17: 0 Linoleic acid-KTTK C18: 2 Margaric acid-RRR C17: 0 Linoleic acid-KTT C18: 2 Margaric acid-RRRRR C17: 0 Linoleic acid-kt C18: 2 Margaric acid-RRRRRRRRR C17: 0 Linoleic acid-EEMQRR C18: 2 Margaric acid-KKKKK C17: 0 Linoleic acid-EEMQR C18: 2 Margaric acid-KKKKKKKKK C17: 0 Linoleic acid-EEMQ C18: 2


16


Stearic acid-LLWIALRKK C18: 0 Linoleic acid-EEM C18: 2 Stearic acid-llwialrkk C18: 0 Linoleic acid-EE C18: 2 Stearic acid-RRR C18: 0 Linoleic acid-KKK C18: 2 Stearic acid-RRRRR C18: 0 Linoleic acid-KK C18: 2 Stearic acid-RRRRRRRRR C18: 0 Linoleic acid-gp C18: 2 Stearic acid-KKKKK C18: 0 Linoleic acid-GQPR C18: 2 Stearic acid-KKKKKKKKK C18: 0 Linoleic acid-gqp C18: 2


17


Behenic acid-LLWIALRKK C22: 0 Linoleic acid-gq C18: 2 Behenic acid-llwialrkk C22: 0 Linoleic acid-LLWIALRKK C18: 2 Behenic acid-RRR C22: 0 Linoleic acid-gpo C18: 2 Behenic acid-RRRRR C22: 0 Linoleic acid-llwialrkk C18: 2 Behenic acid-RRRRRRRRR C22: 0 Linoleic acid-tk C18: 2 Behenic acid-KKKKK C22: 0 Linoleic acid-ttks C18: 2 Behenic acid-KKKKKKKKK C22: 0 Linoleic acid-tks C18: 2


18


Lignoceric acid-LLWIALRKK C24: 0 Linoleic acid-KKKKK C18: 2 Lignoceric acid-llwialrkk C24: 0 Linoleic acid-KKKKKKKKK C18: 2 Lignoceric acid-RRR C24: 0 Linoleic acid-rrr C18: 2 Lignoceric acid-RRRRR C24: 0 Linoleic acid-RRRRR C18: 2 Lignoceric acid-RRRRRRRRR C24: 0 Linoleic acid-RRRRRRRRR C18: 2 Lignoceric acid-KKKKK C24: 0 Lignoceric acid-KKKKKKKKK C24: 0

Type of synthesized fatty acid-oligopeptide complex division name Carbon number division name Carbon number


19


Palmitoleic acid-LLWIALRKK C16: 1





22





Vaccenic acid-AAPV C18: 1 Palmitoleic acid-llwialrkk C16: 1 Vaccenic acid-GHK C18: 1 Palmitoleic acid-rrr C16: 1 Vaccenic acid-KTTKS C18: 1 Palmitoleic acid-RRRRR C16: 1 Vaccenic acid-EEMQRR C18: 1 Palmitoleic acid-RRRRRRRRR C16: 1 Vaccenic acid-KKK C18: 1 Palmitoleic acid-KKKKK C16: 1 Vaccenic acid-LLWIALRKK C18: 1 Palmitoleic acid-KKKKKKKKK C16: 1 Vaccenic acid-GPO C18: 1





20


Elaidic acid-AAPV C18: 1 Vaccenic acid-llwialrkk C18: 1 Elaidic acid-ghk C18: 1 Vaccenic acid-RRR C18: 1 Elaidic acid-KTTKS C18: 1 Vaccenic acid-RRRRR C18: 1 Elaidic acid-EEMQRR C18: 1 Vaccenic acid-RRRRRRRRR C18: 1 Elaidic acid-KKK C18: 1 Vaccenic acid-KKKKK C18: 1 Elaidic acid-LLWIALRKK C18: 1 Vaccenic acid-KKKKKKKKK C18: 1 Elaidic acid-GPO C18: 1





23





Gondoic acid-AAPV C20: 1 Elaidic acid-llwialrkk C18: 1 Gondoic acid-GHK C20: 1 Elaidic acid-rrr C18: 1 Gondoic acid-KTTKS C20: 1 Elaidic acid-RRRRR C18: 1 Gondoic acid-EEMQRR C20: 1 Elaidic acid-RRRRRRRRR C18: 1 Gondoic acid-KKK C20: 1 Elaidic acid-KKKKK C18: 1 Gondoic acid-LLWIALRKK C20: 1 Elaidic acid-KKKKKKKKK C18: 1 Gondoic acid-GPO C20: 1





21





Petroselinic acid-AAPV C18: 1 Gondoic acid-llwialrkk C20: 1 Petroselinic acid-GHK C18: 1 Gondoic acid-RRR C20: 1 Petroselinic acid-KTTKS C18: 1 Gondoic acid-RRRRR C20: 1 Petroselinic acid-EEMQRR C18: 1 Gondoic acid-RRRRRRRRR C20: 1 Petroselinic acid-KKK C18: 1 Gondoic acid-KKKKK C20: 1 Petroselinic acid-LLWIALRKK C18: 1 Gondoic acid-KKKKKKKKK C20: 1 Petroselinic acid-GPO





24





Erucic acid-AAPV C22: 1 Petroselinic acid-llwialrkk C18: 1 Erucic acid-GHK C22: 1 Petroselinic acid-RRR C18: 1 Erucic acid-KTTKS C22: 1 Petroselinic acid-RRRRR C18: 1 Erucic acid-EEMQRR C22: 1 Petroselinic acid-RRRRRRRRR C18: 1 Erucic acid-KKK C22: 1 Petroselinic acid-KKKKK C18: 1 Erucic acid-LLWIALRKK C22: 1 Petroselinic acid-KKKKKKKKK C18: 1 Erucic acid-GPO C22: 1




Erucic acid-llwialrkk C22: 1 Erucic acid-RRR C22: 1 Erucic acid-RRRRR C22: 1 Erucic acid-RRRRRRRRR C22: 1 Erucic acid-KKKKK C22: 1 Erucic acid-KKKKKKKKK C22: 1

Type of synthesized fatty acid-oligopeptide complex division name Carbon number division name Carbon number




25





Nervonic acid-AAPV C24: 1




28






cis-Linoleic acid-AAPV C18: 2 Nervonic acid-GHK C24: 1 cis-Linoleic acid-GHK C18: 2 Nervonic acid-KTTKS C24: 1 cis-Linoleic acid-KTTKS C18: 2 Nervonic acid-EEMQRR C24: 1 cis-Linoleic acid-EEMQRR C18: 2 Nervonic acid-KKK C24: 1 cis-Linoleic acid-KKK C18: 2 Nervonic acid-LLWIALRKK C24: 1 cis-Linoleic acid-LLWIALRKK C18: 2 Nervonic acid-GPO C24: 1 cis-Linoleic acid-GPO C18: 2 Nervonic acid-llwialrkk C24: 1 cis-Linoleic acid-llwialrkk C18: 2 Nervonic acid-RRR C24: 1 cis-Linoleic acid-RRR C18: 2 Nervonic acid-RRRRR C24: 1 cis-Linoleic acid-RRRRR C18: 2 Nervonic acid-RRRRRRRRR C24: 1 cis-Linoleic acid-RRRRRRRRR C18: 2 Nervonic acid-KKKKK C24: 1 cis-Linoleic acid-KKKKK C18: 2 Nervonic acid-KKKKKKKKK C24: 1 cis-Linoleic acid-KKKKKKKKK C18: 2





26





Punicic acid-AAPV C18: 3





29





Docosapentaenoic acid-AAPV C22: 6 Punicic acid-GHK C18: 3 Docosapentaenoic acid-GHK C22: 6 Punicic acid-KTTKS C18: 3 Docosapentaenoic acid-KTTKS C22: 6 Punicic acid-EEMQRR C18: 3 Docosapentaenoic acid-EEMQRR C22: 6 Punicic acid-KKK C18: 3 Docosapentaenoic acid-KKK C22: 6 Punicic acid-LLWIALRKK C18: 3 Docosapentaenoic acid-LLWIALRKK C22: 6 Punicic acid-GPO C18: 3 Docosapentaenoic acid-GPO C22: 6 Punicic acid-llwialrkk C18: 3 Docosapentaenoic acid-llwialrkk C22: 6 Punicic acid-RRR C18: 3 Docosapentaenoic acid-RRR C22: 6 Punicic acid-RRRRR C18: 3 Docosapentaenoic acid-RRRRR C22: 6 Punicic acid-RRRRRRRRR C18: 3 Docosapentaenoic acid-RRRRRRRRR C22: 6 Punicic acid-KKKKK C18: 3 Docosapentaenoic acid-KKKKK C22: 6 Punicic acid-KKKKKKKKK C18: 3 Docosapentaenoic acid-KKKKKKKKK C22: 6




27





Eleostearic acid-AAPV C18: 3





30





Ricinoleic acid-AAPV C18: 1 Eleostearic acid-GHK C18: 3 Ricinoleic acid-GHK C18: 1 Eleostearic acid-KTTKS C18: 3 Ricinoleic acid-KTTKS C18: 1 Eleostearic acid-EEMQRR C18: 3 Ricinoleic acid-EEMQRR C18: 1 Eleostearic acid-KKK C18: 3 Ricinoleic acid-KKK C18: 1 Eleostearic acid-LLWIALRKK C18: 3 Ricinoleic acid-LLWIALRKK C18: 1 Eleostearic acid-GPO C18: 3 Ricinoleic acid-GPO C18: 1 Eleostearic acid-llwialrkk C18: 3 Ricinoleic acid-llwialrkk C18: 1 Eleostearic acid-RRR C18: 3 Ricinoleic acid-RRR C18: 1 Eleostearic acid-RRRRR C18: 3 Ricinoleic acid-RRRRR C18: 1 Eleostearic acid-RRRRRRRRR C18: 3 Ricinoleic acid-RRRRRRRRR C18: 1 Eleostearic acid-KKKKK C18: 3 Ricinoleic acid-KKKKK C18: 1 Eleostearic acid-KKKKKKKKK C18: 3 Ricinoleic acid-KKKKKKKKK C18: 1

Type of synthesized fatty acid-oligopeptide complex division name Carbon number division name Carbon number



31



Palmitic acid-KTTKS C18: 1




32




Capric acid-KTTKS C18: 1 KTTKS-Palmitic acid C18: 1 KTTKS-Capric acid C18: 1 Pmitmitic acid (KTTKS) C18: 1 KTTKS (Capric acid) C18: 1 Palmitic acid-KTTK C18: 1 Capric acid-KTTK C18: 1 KTTK-Palmitic acid C18: 1 KTTK-Capric acid C18: 1 Pmitmitic acid (KTTK) C18: 1 Capric acid (KTTK) C18: 1 Palmitic acid-RRRRR C18: 1 Capric acid-RRRRR C18: 1 RRRRR-Palmitic acid C18: 1 RRRRR-Capric acid C18: 1 Palmitic acid-KKKKK C18: 1 Capric acid-KKKKK C18: 1 KKKKK-Palmitic acid C18: 1 KKKKK-Capric acid C18: 1 KKKKK (Palmitic acid) C18: 1 KKKKK (Capric acid) C18: 1

Type of synthesized fatty acid-oligopeptide complex division name division name







33






P1 LLWIALRKK-NH ₂ P15 LLWIALRKK-COOH P2 LLWIALRK K (C6: 0) -NH ₂ P16 LLWIALRK K (C6: 0) -COOH P3 LLWIALRK K (C8: 0) -NH ₂ P17 LLWIALRK K (C8: 0) -COOH P4 LLWIALRK K (C10: 0) -NH ₂ P18 LLWIALRK K (C10: 0) -COOH P5 LLWIALRK K (C12: 0) -NH ₂ P19 LLWIALRK K (C12: 0) -COOH P6 LLWIALRK K (C14: 0) -NH ₂ P20 LLWIALRK K (C14: 0) -COOH P7 LLWIALRK K (C16: 0) -NH ₂ P21 LLWIALRK K (C16: 0) -COOH P8 llwialrkk-NH ₂ P22 llwialrkk- COOH P9 llwialrk k (C6: 0) -NH ₂ P23 llwialrk k (C6: 0) -COOH P10 llwialrk k (C8: 0) -NH ₂ P24 llwialrk k (C8: 0) -COOH P11 llwialrk k (C10: 0) -NH ₂ P25 llwialrk k (C10: 0) -COOH P12 llwialrk k (C12: 0) -NH ₂ P26 llwialrk k (C12: 0) -COOH P13 llwialrk k (C14: 0) -NH ₂ P27 llwialrk k (C14: 0) -COOH P14 llwialrk k (C16: 0) -NH ₂ P28 llwialrk k (C16: 0) -COOH

Example  2. Complex Tablets

The composite preparations synthesized in Example 1 were dissolved in distilled water to which 10% (v / v) acetonitrile was added, and then purified by high performance liquid chromatography (HPLC) under gradient conditions (FIG. 2). To 4), lyophilization to obtain the desired complex. HPLC conditions and gradient conditions of the separation phase are shown in Tables 11 and 12 below. And the molecular weight of the purified composite preparations were measured using MALDI-TOF-MASS, the molecular weight measurement method is shown in Table 13, the molecular weight measurement results are shown in Table 14 to Table 23.

HPLC conditions device Shimadzu 8A (5cm × 25cm) / 8A (prep (dia 5cm)) Solvent composition Buffer A = 0.05% TFA / acetonitrile, Buffer B = 0.05% TFA / H ₂ O Flow rate 14 ml / min column Silicagel C18 reverse phase Absorbance 230 nm Separation condition Gradient

Gradient conditions Minutes H ₂ 0 (%) Acetonitrile (%) 0-12 89 → 77 11 → 23 12-13 77 → 5 23 → 95 13-18 5 → 5 95 → 95 18 ~ 19 5 → 89 95 → 11 19-27 89 → 99 11 → 11

Molecular weight measuring method -Device Name: SHIMADZU MALDI-TOF
Mix 5 μL of the sample with 5 μL of the matrix solution and load 0.5 μL of the mixture onto the target plate.
-Then measure according to the manual of the mass spectrometer.
Where the matrix solution is a saturated solution of alpha-cyanohydroxycinnamic acid in 0.1% Trifluoroacetic acid (TFA) acetonitrile / water = 1/1 (v / v).

Check molecular weight division Complex sequence Composite location
( Peptide  standard) theory
Molecular Weight Actual measurement
Molecular Weight











One















One Capric acid-AAPV








N-term 510.7 511.6 2 Capric acid-AAP 410.6 409.8 3 Capric acid-AA 313.5 314.4 4 Capric acid-gh 366.5 366.4 5 Capric acid-ghk 494.6 493.8 6 Capric acid-KTTKS 717.9 718.4 7 Capric acid-KTTK 630.8 631.8 8 Capric acid-KTT 502.7 500.6 9 Capric acid-KT 401.6 400.5 10 Capric acid-EEMQRR 1002.2 1003.1 11 Capric acid-EEMQR 846.0 847.0 12 Capric acid-EEMQ 689.8 690.8 13 Capric acid-EEM 561.7 562.7 14 Capric acid-ee 430.5 432.4 15 Capric acid-KKK 556.8 557.4 16 Capric acid-KK 428.6 429.6 17 Capric acid-gp 326.4 327.4 18 Capric acid-GQPR 610.8 611.5 19 Capric acid-GQP 454.6 456.5 20 Capric acid-gq 357.5 359.4 21 LLWIALRK K (Capric acid) K (side chain) 1294.7 1295.5 22 Capric acid-GPO N-term 440.6 441.3 23 llwialrk k (Capric acid) K (side chain) 1294.7 1295.1 24 Capric acid-TK


N-term
401.6 402.5 25 Capric acid-TTKS 589.7 590.7 26 Capric acid-TKS 488.6 490.6 27 Capric acid-KKKKK 813.1 814.1 28 Capric acid-KKKKKKKKK 1325.8 1326.8 29 Capric acid-RRR 640.8 642.8 30 Capric acid-RRRRR 953.2 954.2 31 Capric acid-RRRRRRRRR 1578.0 1579.9






2







One Palmitic acid-AAPV






N-term
594.8 595.8 2 Palmitic acid-AAP 494.7 496.7 3 Palmitic acid-AA 397.6 397.7 4 Palmitic acid-GHK 578.8 578.8 5 Palmitic acid-gh 450.6 452.6 6 Palmitic acid-KTTKS 802.1 803.0 7 Palmitic acid-KTTK 715.0 715.9 8 Palmitic acid-KTT 586.8 588.8 9 Palmitic acid-KT 485.7 486.6 10 Palmitic acid-EEMQRR 1086.3 1088.3 11 Palmitic acid-EEMQR 930.2 930.1 12 Palmitic acid-EEMQ 774.0 774.9 13 Palmitic acid-EEM 645.8 644.8 14 Palmitic acid-EE 514.6 516.6 15 Palmitic acid-KKK 640.9 642.9 16 Palmitic acid-KK 512.8 513.8 17 Palmitic acid-gp 410.6 411.2

K side chain; Fatty acid binding to oligopeptide of SEQ ID 30 (LLWIALRKK, llwialrkk) 9th K amino acid (underlined).

Check molecular weight division Complex sequence Composite location
( Peptide  standard) theory
Molecular Weight Actual measurement
Molecular Weight




2






18 Palmitic acid-GQPR
N-term
694.9 695.9 19 Palmitic acid-GQP 538.7 536.7 20 Palmitic acid-gq 441.6 443.6 21 LLWIALRK K (Palmitic acid) K (side chain) 1378.9 1379.9 22 Palmitic acid-GPO N-term 524.7 525.5 23 llwialrk k (Palmitic acid) K (side chain) 1378.9 1379.8 24 Palmitic acid-TK


N-term 485.7 486.6 25 Palmitic acid-TTKS 673.9 674.8 26 Palmitic acid-TKS 572.8 574.7 27 Palmitic acid-KKKKK 897.3 898.2 28 Palmitic acid-KKKKKKKKK 1410.0 1409.8 29 Palmitic acid-RRR 725.0 722.9 30 Palmitic acid-RRRRR 1037.4 1038.4 31 Palmitic acid-RRRRRRRRR 1662.1 1662.8













3

















One Oleic acid-AAPV














N-term














620.9 621.8 2 Oleic acid-AAP 520.8 521.6 3 Oleic acid-AA 423.7 423.9 4 Oleic acid-A 353.6 354.5 5 Oleic acid-GHK 604.9 605.4 6 Oleic acid-GH 476.7 478.5 7 Oleic acid-g 339.5 340.2 8 Oleic acid-KTTKS 828.1 829.4 9 Oleic acid-KTTK 741.0 742.3 10 Oleic acid-KTT 612.9 613.5 11 Oleic acid-KT 511.8 512.9 12 Oleic acid-EEMQRR 1112.4 1113.5 13 Oleic acid-EEMQR 956.2 957.2 14 Oleic acid-EEMQ 800.0 800.9 15 Oleic acid-EEM 671.9 672.8 16 Oleic acid-EE 540.7 540.7 17 Oleic acid-E 411.6 412.6 18 Oleic acid-KKK 667.0 667.9 19 Oleic acid-KK 538.8 539.6 20 Oleic acid-K 410.7 411.2 21 Oleic acid-GP 436.7 437.4 22 Oleic acid-GQPR 721.0 722.1 23 Oleic acid-GQP 564.8 565.3 24 Oleic acid-gq 467.7 468.4 25 Oleic acid-LLWIALRKK 1404.9 1404.5 26 Oleic acid-GPO 550.8 551.4 27 Oleic acid-llwialrkk 1404.9 1405.6 28 Oleic acid-TK 511.8 513.1 29 Oleic acid-TTKS 699.9 697.1 30 Oleic acid-TKS 598.8 599.8 31 Oleic acid-KKKKK 923.4 923.7 32 Oleic acid-KKKKKKKKK 1436.0 1437.1 33 Oleic acid-RRR 751.0 751.8 34 Oleic acid-RRRRR 1063.4 1063.9 35 Oleic acid-RRRRRRRRR 1688.2 1689.4

Check molecular weight division Complex sequence Composite location
( Peptide  standard) theory
Molecular Weight Actual measurement
Molecular Weight













4












One Linoleic acid-AAPV













N-term















618.9 619.4 2 Linoleic acid-AAP-OH 518.7 519.4 3 Linoleic acid-AA-OH 421.6 422.4 4 Linoleic acid-ghk 602.8 603.6 5 Linoleic acid-gh 474.7 475.1 6 Linoleic acid-KTTKS 826.1 827.1 7 Linoleic acid-KTTK 739.0 739.8 8 Linoleic acid-KTT 610.8 611.6 9 Linoleic acid-kt 509.7 511.0 10 Linoleic acid-EEMQRR 1110.4 1111.5 11 Linoleic acid-EEMQR 954.2 955.1 12 Linoleic acid-EEMQ 798.0 799.0 13 Linoleic acid-EEM 669.9 670.4 14 Linoleic acid-EE 538.7 539.5 15 Linoleic acid-KKK 665.0 666.1 16 Linoleic acid-KK 536.8 536.8 17 Linoleic acid-gp 434.6 435.5 18 Linoleic acid-GQPR 718.9 718.7 19 Linoleic acid-gqp 562.8 563.7 20 Linoleic acid-gq 465.6 466.6 21 Linoleic acid-LLWIALRKK 1402.9 1403.6 22 Linoleic acid-gpo 548.8 549.2 23 Linoleic acid-llwialrkk 1402.9 1403.7 24 Linoleic acid-tk 509.7 510.4 25 Linoleic acid-ttks 697.9 698.8 26 Linoleic acid-tks 596.8 597.4 27 Linoleic acid-KKKKK 921.3 922.6 28 Linoleic acid-KKKKKKKKK 1434.0 1435.1 29 Linoleic acid-rrr 749.0 751.0 30 Linoleic acid-RRRRR 1061.4 1062.1 31 Linoleic acid-RRRRRRRRR 1686.1 1687.2




5





One Caprylic acid-AAPV

N-term

482.6 483.3 2 Caprylic acid-GHK 466.6 467.4 3 Caprylic acid-KTTKS 689.9 691.0 4 Caprylic acid-EEMQRR 974.2 975.1 5 Caprylic acid-KKK 528.7 529.3 6 LLWIALRK K (Caprylic acid) K (side chain) 1266.7 1266.9 7 Caprylic acid-GPO N-term 412.5 413.5 8 llwialrk k (Caprylic acid) K (side chain) 1266.7 1266.9 9 Caprylic acid-RRR

N-term
612.8 613.5 10 Caprylic acid-RRRRR 925.2 925.9 11 Caprylic acid-RRRRRRRRR 1549.9 1550.0 12 Caprylic acid-KKKKK 785.1 786.7 13 Caprylic acid-KKKKKKKKK 1297.8 1298.8

Check molecular weight division Complex sequence Composite location
( Peptide  standard) theory
Molecular Weight Actual measurement
Molecular Weight





6





One Lauric acid-AAPV

N-term

538.7 539.5 2 Lauric acid-ghk 522.7 523.5 3 Lauric acid-KTTKS 746.0 747.0 4 Lauric acid-EEMQRR 1030.3 1030.8 5 Lauric acid-KKK 584.8 585.7 6 LLWIALRK K (Lauric acid) K (side chain) 1322.8 132.6 7 Lauric acid-GPO N-term 468.6 469.3 8 llwialrk k (Lauric acid) K (side chain) 1322.8 1323.6 9 Lauric acid-RRR

N-term

668.9 669.6 10 Lauric acid-RRRRR 981.3 982.2 11 Lauric acid-RRRRRRRRR 1606.0 1607.1 12 Lauric acid-KKKKK 841.2 842.0 13 Lauric acid-KKKKKKKKK 1353.9 1354.7





7





One Myristic acid-AAPV

N-term

566.8 567.3 2 Myristic acid-GHK 550.8 551.2 3 Myristic acid-KTTKS 774.0 775.1 4 Myristic acid-EEMQRR 1058.3 1059.2 5 Myristic acid-KKK 612.9 612.9 6 LLWIALRK K (Myristic acid) K (side chain) 1350.8 1351.6 7 Myristic acid-GPO N-term 496.7 497.6 8 llwialrk k (Myristic acid) K (side chain) 1350.8 1351.4 9 Myristic acid-RRR

N-term

696.9 670.2 10 Myristic acid-RRRRR 1009.3 1010.2 11 Myristic acid-RRRRRRRRR 1634.1 1635.1 12 Myristic acid-KKKKK 869.3 870.2 13 Myristic acid-KKKKKKKKK 1381.9 1382.4





8





One Margaric acid-AAPV





N-term





608.9 609.3 2 Margaric acid-GHK 692.8 593.3 3 Margaric acid-KTTKS 816.1 817.0 4 Margaric acid-EEMQRR 1100.4 1101.0 5 Margaric acid-KKK 655.0 655.9 6 Margaric acid-LLWIALRKK 1392.9 1393.4 7 Margaric acid-GPO 538.8 539.6 8 Margaric acid-llwialrkk 1392.9 1374.0 9 Margaric acid-RRR 739.0 741.2 10 Margaric acid-RRRRR 1051.4 1052.1 11 Margaric acid-RRRRRRRRR 1676.1 1677.3 12 Margaric acid-KKKKK 911.3 912.2 13 Margaric acid-KKKKKKKKK 1424.0 1425.2

Check molecular weight division Complex sequence Composite location
( Peptide  standard) theory
Molecular Weight Actual measurement
Molecular Weight





9





One Stearic acid-AAPV




N-term






622.9 623.8 2 Stearic acid-GHK 06.9 07.5 3 Stearic acid-KTTKS 30.1 30.7 4 Stearic acid-EEMQRR 114.4 115.2 5 Stearic acid-KKK 69.0 70.2 6 Stearic acid-LLWIALRKK 406.9 407.8 7 Stearic acid-GPO 52.8 53.8 8 Stearic acid-llwialrkk 406.9 407.2 9 Stearic acid-RRR 53.1 54.0 10 Stearic acid-RRRRR 065.4 065.9 11 Stearic acid-RRRRRRRRR 690.2 691.1 12 Stearic acid-KKKKK 25.4 26.0 13 Stearic acid-KKKKKKKKK 438.1 439.0





10





One Behenic acid-AAPV





N-term





679.0 79.4 2 Behenic acid-GHK 63.0 63.8 3 Behenic acid-KTTKS 86.2 87.1 4 Behenic acid-EEMQRR 170.5 171.5 5 Behenic acid-KKK 25.1 26.0 6 Behenic acid-LLWIALRKK 463.0 463.7 7 Behenic acid-GPO 08.9 09.4 8 Behenic acid-llwialrkk 463.0 463.6 9 Behenic acid-RRR 09.2 09.9 10 Behenic acid-RRRRR 121.5 122.6 11 Behenic acid-RRRRRRRRR 746.3 747.2 12 Behenic acid-KKKKK 81.5 82.5 13 Behenic acid-KKKKKKKKK 494.2 494.5





11





One Lignoceric acid-AAPV





N-term





707.1 707.6 2 Lignoceric acid-GHK 691.0 492.5 3 Lignoceric acid-KTTKS 914.3 915.2 4 Lignoceric acid-EEMQRR 1198.6 1199.2 5 Lignoceric acid-KKK 753.2 754.2 6 Lignoceric acid-LLWIALRKK 1491.1 1492.2 7 Lignoceric acid-GPO 637.0 638.0 8 Lignoceric acid-llwialrkk 1491.1 1492.2 9 Lignoceric acid-RRR 837.2 838.2 10 Lignoceric acid-RRRRR 1149.6 1151.2 11 Lignoceric acid-RRRRRRRRR 1774.3 1775.1 12 Lignoceric acid-KKKKK 1009.5 1010.0 13 Lignoceric acid-KKKKKKKKK 1522.2 1523.1

Check molecular weight division Complex sequence Composite location
( Peptide  standard) theory
Molecular Weight Actual measurement
Molecular Weight





12





One Palmitoleic acid-AAPV





N-term




592.8 593.1 2 Palmitoleic acid-ghk 576.8 577.4 3 Palmitoleic acid-KTTKS 800.1 801.0 4 Palmitoleic acid-EEMQRR 1084.4 1085.4 5 Palmitoleic acid-KKK 638.9 639.4 6 Palmitoleic acid-LLWIALRKK 1376.9 1377.6 7 Palmitoleic acid-GPO 522.7 523.0 8 Palmitoleic acid-llwialrkk 1376.9 1377.3 9 Palmitoleic acid-rrr 723.0 724.0 10 Palmitoleic acid-RRRRR 1035.4 1036.0 11 Palmitoleic acid-RRRRRRRRR 1660.1 1661.0 12 Palmitoleic acid-KKKKK 895.3 896.0 13 Palmitoleic acid-KKKKKKKKK 1408.0 1409.0





13





One Elaidic acid-AAPV





N-term





620.9 621.2 2 Elaidic acid-ghk 604.8 605.2 3 Elaidic acid-KTTKS 828.1 829.0 4 Elaidic acid-EEMQRR 1112.4 1113.2 5 Elaidic acid-KKK 667.0 667.9 6 Elaidic acid-LLWIALRKK 1404.9 1405.3 7 Elaidic acid-GPO 550.8 551.3 8 Elaidic acid-llwialrkk 1404.9 1405.4 9 Elaidic acid-rrr 751.0 752.0 10 Elaidic acid-RRRRR 1063.2 1064.1 11 Elaidic acid-RRRRRRRRR 1668.2 1689.4 12 Elaidic acid-KKKKK 923.3 925.0 13 Elaidic acid-KKKKKKKKK 1436.0 1437.2





14

One Petroselinic acid-AAPV





N-term
620.9 621.7 2 Petroselinic acid-GHK 604.8 605.9 3 Petroselinic acid-KTTKS 828.1 823.8 4 Petroselinic acid-EEMQRR 1112.4 1113.3 5 Petroselinic acid-KKK 667.0 337.9 6 Petroselinic acid-LLWIALRKK 1404.9 1405.2 7 Petroselinic acid-GPO 550.9 551.6 8 Petroselinic acid-llwialrkk 1404.9 1405.3 9 Petroselinic acid-RRR 751.0 752.0 10 Petroselinic acid-RRRRR 1063.4 1064.2 11 Petroselinic acid-RRRRRRRRR 1688.2 1689.0 12 Petroselinic acid-KKKKK 823.3 924.1 13 Petroselinic acid-KKKKKKKKK 1436.0 1437.3

Check molecular weight division Complex sequence Composite location
( Peptide  standard) theory
Molecular Weight Actual measurement
Molecular Weight





15





One Vaccenic acid-AAPV





N-term





620.9 621.1 2 Vaccenic acid-GHK 604.8 405.0 3 Vaccenic acid-KTTKS 828.1 829.1 4 Vaccenic acid-EEMQRR 1112.4 1112.7 5 Vaccenic acid-KKK 667.0 667.8 6 Vaccenic acid-LLWIALRKK 1404.9 1405.2 7 Vaccenic acid-GPO 550.8 551.4 8 Vaccenic acid-llwialrkk 1404.9 1405.6 9 Vaccenic acid-RRR 751.0 752.2 10 Vaccenic acid-RRRRR 1063.4 1064.1 11 Vaccenic acid-RRRRRRRRR 1688.2 1689.7 12 Vaccenic acid-KKKKK 923.3 923.9 13 Vaccenic acid-KKKKKKKKK 1436.0 1437.2





16





One Gondoic acid-AAPV





N-term





648.9 649.2 2 Gondoic acid-GHK 632.9 633.8 3 Gondoic acid-KTTKS 856.2 857.3 4 Gondoic acid-EEMQRR 1140.5 1140.9 5 Gondoic acid-KKK 695.0 496.0 6 Gondoic acid-LLWIALRKK 1433.0 1433.8 7 Gondoic acid-GPO 578.8 579.3 8 Gondoic acid-llwialrkk 1433.0 1434.2 9 Gondoic acid-RRR 779.1 780.0 10 Gondoic acid-RRRRR 1091.5 1092.7 11 Gondoic acid-RRRRRRRRR 1716.2 1717.8 12 Gondoic acid-KKKKK 951.4 952.6 13 Gondoic acid-KKKKKKKKK 1464.1 1465.2





17





One Erucic acid-AAPV





N-term





677.0 677.9 2 Erucic acid-GHK 661.0 662.1 3 Erucic acid-KTTKS 884.2 885.3 4 Erucic acid-EEMQRR 1168.5 1169.2 5 Erucic acid-KKK 723.1 724.1 6 Erucic acid-LLWIALRKK 1461.0 1462.3 7 Erucic acid-GPO 606.9 607.2 8 Erucic acid-llwialrkk 1461.0 1461.7 9 Erucic acid-RRR 807.1 807.6 10 Erucic acid-RRRRR 1119.5 1120.4 11 Erucic acid-RRRRRRRRR 1744.3 1744.8 12 Erucic acid-KKKKK 979.5 980.4 13 Erucic acid-KKKKKKKKK 1492.1 1493.2

Check molecular weight division Complex sequence Composite location
( Peptide  standard) theory
Molecular Weight Actual measurement
Molecular Weight





18





One Nervonic acid-AAPV





N-term





705.0 705.9 2 Nervonic acid-GHK 689.0 689.6 3 Nervonic acid-KTTKS 912.3 912.7 4 Nervonic acid-EEMQRR 1196.6 1197.5 5 Nervonic acid-KKK 751.2 752.3 6 Nervonic acid-LLWIALRKK 1489.1 1490.1 7 Nervonic acid-GPO 635.0 636.2 8 Nervonic acid-llwialrkk 1489.1 1490.8 9 Nervonic acid-RRR 835.2 836.2 10 Nervonic acid-RRRRR 1147.6 1148.2 11 Nervonic acid-RRRRRRRRR 1772.3 1774.1 12 Nervonic acid-KKKKK 1007.5 1009.2 13 Nervonic acid-KKKKKKKKK 1520.2 1520.9





19





One cis-Linolenic acid-AAPV





N-term





618.9 619.2 2 cis-Linolenic acid-GHK 602.8 603.2 3 cis-Linolenic acid-KTTKS 826.1 827.2 4 cis-Linolenic acid-EEMQRR 1110.4 1110.8 5 cis-Linolenic acid-KKK 665.0 66.0 6 cis-Linolenic acid-LLWIALRKK 1402.9 1403.4 7 cis-Linolenic acid-GPO 548.8 549.5 8 cis-Linolenic acid-llwialrkk 1402.9 1403.7 9 cis-Linolenic acid-RRR 749.0 750.2 10 cis-Linolenic acid-RRRRR 1061.4 1062.5 11 cis-Linolenic acid-RRRRRRRRR 1686.1 1687.1 12 cis-Linolenic acid-KKKKK 921.3 922.3 13 cis-Linolenic acid-KKKKKKKKK 1434.0 1435.5




20





One Punicic acid-AAPV





N-term





616.9 617.0 2 Punicic acid-GHK 600.8 600.9 3 Punicic acid-KTTKS 824.1 824.4 4 Punicic acid-EEMQRR 1108.4 1109.0 5 Punicic acid-KKK 663.0 664.2 6 Punicic acid-LLWIALRKK 1400.9 1401.3 7 Punicic acid-GPO 546.8 547.4 8 Punicic acid-llwialrkk 1400.9 1401.4 9 Punicic acid-RRR 747.0 747.7 10 Punicic acid-RRRRR 1059.4 1060.3 11 Punicic acid-RRRRRRRRR 1684.1 1685.2 12 Punicic acid-KKKKK 919.3 920.0 13 Punicic acid-KKKKKKKKK 1432.0 1433.4

Check molecular weight division Complex sequence Composite location
( Peptide  standard) theory
Molecular Weight Actual measurement
Molecular Weight





21





One Eleostearic acid-AAPV





N-term





616.9 617.7 2 Eleostearic acid-GHK 600.8 601.3 3 Eleostearic acid-KTTKS 824.1 825.4 4 Eleostearic acid-EEMQRR 1108.4 1109.5 5 Eleostearic acid-KKK 663.0 664.0 6 Eleostearic acid-LLWIALRKK 1400.9 1401.0 7 Eleostearic acid-GPO 546.8 547.8 8 Eleostearic acid-llwialrkk 1400.9 1401.3 9 Eleostearic acid-RRR 747.0 748.2 10 Eleostearic acid-RRRRR 1059.4 1060.2 11 Eleostearic acid-RRRRRRRRR 1684.1 1684.9 12 Eleostearic acid-KKKKK 919.3 920.1 13 Eleostearic acid-KKKKKKKKK 1432.0 1433.4





22





One Docosapentaenoic acid-AAPV





N-term





668.4 669.0 2 Docosapentaenoic acid-GHK 652.4 653.2 3 Docosapentaenoic acid-KTTKS 875.7 876.5 4 Docosapentaenoic acid-EEMQRR 1159.9 1160.6 5 Docosapentaenoic acid-KKK 714.5 715.2 6 Docosapentaenoic acid-LLWIALRKK 1452.5 1453.4 7 Docosapentaenoic acid-GPO 598.3 599.4 8 Docosapentaenoic acid-llwialrkk 1452.5 1453.2 9 Docosapentaenoic acid-RRR 798.6 799.3 10 Docosapentaenoic acid-RRRRR 1111.0 111.8 11 Docosapentaenoic acid-RRRRRRRRR 1735.7 1736.2 12 Docosapentaenoic acid-KKKKK 970.9 971.5 13 Docosapentaenoic acid-KKKKKKKKK 1492.6 1484.2





23





One Ricinoleic acid-AAPV





N-term





618.9 619.1 2 Ricinoleic acid-GHK 602.8 603.4 3 Ricinoleic acid-KTTKS 826.1 837.0 4 Ricinoleic acid-EEMQRR 1110.4 1110.9 5 Ricinoleic acid-KKK 665.0 665.9 6 Ricinoleic acid-LLWIALRKK 1402.9 1403.4 7 Ricinoleic acid-GPO 548.8 549.3 8 Ricinoleic acid-llwialrkk 1402.9 1403.6 9 Ricinoleic acid-RRR 749.0 749.7 10 Ricinoleic acid-RRRRR 1061.4 1062.2 11 Ricinoleic acid-RRRRRRRRR 1686.1 1687.0 12 Ricinoleic acid-KKKKK 921.3 922.2 13 Ricinoleic acid-KKKKKKKKK 1434.0 1435.0

Check molecular weight division Complex sequence Composite location
( Peptide  standard) theory
Molecular Weight Actual measurement
Molecular Weight


24


One KTTKS-ethylene glycol-Palmitic acid C-term 846.1 846.5 2 KTT K S (Palmitic acid) K (side chain) 802.1 802.5 3 KTTK-ethylene glycol-Palmitic acid C-term 759.0 760.1 4 KTT K (Palmitic acid) K (side chain) 715.0 716.0 5 RRRRR-ethylene glycol-Palmitic acid C-term
1081.4 1081.9 6 KKKKK-ethylene glycol-Palmitic acid 1019.4 1020.3 7 KKKK K (Palmtic acid) K (side chain) 959.4 960.0


25


One KTTKS-ethylene glycol-Capric acid C-term 762.0 762.8 2 KTT K S (Capric acid) K (side chain) 717.9 717.9 3 KTTK-ethylene glycol-Capric acid C-term 674.9 675.4 4 KTT K (Capric acid) K (side chain) 630.8 631.1 5 RRRRR-ethylene glycol-Capric acid C-term
997.3 997.9 6 KKKKK-ethylene glycol-Capric acid 857.2 858.0 7 KKKK K (Capric acid) K (side chain) 813.1 813.9

Example  3. HaCaT  Cytotoxicity Analysis of Ultrafine Dust in Cells

To confirm the cytotoxicity of the fatty acid-amino acid complex and fatty acid-oligopeptide (SEQ ID NOS: 1 to 30) complex, dilute in DMEM (10% FBS) medium so that the cell number is 2 × 10 ⁴ cells / ml and in a 96-well culture vessel. After 100 μl aliquots were incubated for 16-20 hours until the cells adhered to the plate surface. The medium was removed, and each complex material was diluted to 10 μM concentration with DMEM medium, treated with 100 μl of cells, incubated in 37 ° C., 5% CO ₂ incubator for 24 hours, and then the medium was removed again. Dispense 100 μl of a 10-fold dilution of MTT (5 mg / ml in PBS) reagent into DMEM medium, react for 3 to 4 hours, remove the MTT reagent, and dispense 100 μl of DMSO (dimethyl sulfoxide) in the incubator. After 30 minutes of reaction in the DMEM medium treatment group, 0.2% DMSO / DMEM treatment group (negative control group), 10% DMSO / DMEM treatment group (positive control group) by measuring the absorbance (540 nm) and in Tables 4 to 10 above Cell viability of the group treated with the conjugated amino acid or oligopeptide and fatty acid conjugate was analyzed.

As a result, the cell survival rate of the complex-treated group was 100 ± 4%, which was similar to that of the negative control group, compared to the negative control group with 100% cell viability. It was confirmed that it was ˜25% (holding result not shown). Through this amino acid of the present invention; Or the N-terminal, C-terminal or side chain of the oligopeptide; fatty acid is conjugated to the complex was found to be non-toxic to skin cells.

Example  4. Amino Acids for Ultrafine Dust or Oligopeptides  fatty acid Cytoprotective effect of the complex ( HaCaT  cell)

To determine if the fatty acid-amino acid complex and fatty acid-oligopeptide (SEQ ID NOS: 1-30) complex can reduce the increased cytotoxicity caused by ultrafine dust, ultrafine dust on human skin-derived keratinocytes (HaCaT cells) Diesel exhaust particles, DEP) And the complexes were analyzed for cytotoxicity via MTT assay. The DEP (fine dust having a diameter of 2.5 μm or less, PM2.5) is purchased from the National Institute for Environmental Studies (Japan), and is a 4JB1 type 2740 cc four-cylinder direct injection diesel engine (Isuzu Automobile Co., Japan). Was collected by a glass fiber filter operating at 1500 rpm load conditions and 10 torque (10 kg / ml) conditions.

Cells were diluted in DMEM (10% FBS) medium so that the number of cells was 2 × 10 ⁴ cells / ml, 100 μl of the cells were dispensed into 96-well culture vessels, and cultured for 16-20 hours until the cells adhered to the plate surface. After the incubation was completed, the medium was removed, and 100 μl of a solution containing 10 μM complex and 50 μg / ml ultrafine dust in each well was incubated in a 37 ° C., 5% CO ₂ incubator for 24 hours. The medium was removed again. Thereafter, cell viability was confirmed by MTT assay in the same manner as in Example 3.

As a result, it was confirmed that the cell viability was significantly increased in the group treated with ultrafine dust and complexes combined with ultrafine dust and amino acids or oligopeptides and fatty acids compared to the group treated with only ultrafine dust (FIGS. 5 to 15)

 Through this, it was found that the complexes of fatty acids bound to the amino acids or oligopeptides of the present invention can effectively protect skin cells by agglomerating ultrafine dust.

Example  5. Cytotoxicity Analysis of Ultrafine Dust in A549 Cells

After ultrafine dust treatment on human lung cancer epithelial cell line (A549), cytotoxicity was analyzed by MTT assay. The MTT assay was performed in the same manner as in Example 3, and cultured A549 cells were treated with ultrafine dust at concentrations of 50, 60, 70, 80, 90, 100, 110, 120 and 200 μg / ml.

As a result, as the concentration of ultrafine dust increased, the cell viability decreased, and it was confirmed that only about 20% of cells survived in the highest concentration of 200 μg / ml fine dust (FIG. 16).

Example  6. For ultrafine dust Peptides and  Analysis of Cytoprotective Effect of Fatty Acid Complex (A549 Cell)

Cytotoxicity was analyzed by MTT assay to determine whether the fatty acid-oligopeptide (saturated fatty acid binding to the amino acid sequence side chain of SEQ ID NO: 30) complex can reduce the cytotoxicity caused by ultrafine dust. MTT assay was performed in the same manner as in Example 3.

As a result, the cell survival rate was increased in the group treated with fatty acid-oligopeptide complexes compared to the group treated with ultrafine dust alone (approximately 40%). Cell viability was significantly increased in the peptide complex. Among them, the complex (P4, P11, P18, P25) combined with capric acid (C10) showed more than 70% cell viability. It was confirmed to reduce toxicity (FIG. 17).

Example  7. Fatty acids using silver staining Oligopeptide  Coagulation Effect Analysis of Complex and Ultrafine Dust

Ultrafine dust and fatty acid-oligopeptide complex P11 (capric acid binding to the side chain of the amino acid sequence of SEQ ID NO: 30) simultaneously on a 20% SDS-PAGE gel and the amount of complex that is not aggregated with ultrafine dust by silver staining It was confirmed.

DMEM medium was treated with 100 μg / ml ultrafine dust alone or simultaneously with ultrafine dust and P11 complexes at 10, 50 and 100 μM concentrations and then incubated in 37 ° C., 5% CO ₂ incubator for 24 hours and centrifugal force. Supernatant samples were prepared by centrifugation at 10,000 × g for 1 hour. Then 15 μl of the supernatant was loaded on a 20% SDS-PAGE gel and subjected to electrophoresis. The silver staining process was performed using Elpis Biotech's kit (PeptiGel ™, ELPISBIOTECH, EBA-1053) as follows. After the electrophoresis was completed, the gel was reacted with a solution of 30% ethanol and 10% acetic acid for 1 hour to be immobilized, and washed twice with distilled water. After soaking in solution A for 1 minute, washed twice with distilled water, and soaking the gel in solution B containing formaldehyde 20-30 minutes. Finally, the gel was washed twice with distilled water for 1 minute and then immersed in solution C and waited until a band appeared. Then, the reaction was stopped using a fixed solution.

As a result, in the group treated with ultrafine dust and complex P11 at the same time, the band intensity was slightly decreased compared with the group treated with complex P11 alone. It was found to be reduced (FIG. 18). Through this, it was found that the complexes in which the fatty acid is bound to the oligopeptide of the present invention can remove ultrafine dust by aggregation with ultrafine dust.

Example  8. Fatty Acid- Using Comparison of Permeation Rate of Fine Dust Particles in Masks Oligopeptide  Dust in the complex Capture (Particle removal) efficiency analysis

In order to check whether the fatty acid-oligopeptide complex moves to increase the filter efficiency of the mask by adsorbing the ultrafine dust, the efficiency of dust collection (particle removal) was analyzed. .

Dust collection efficiency evaluation and calculation formula 1) Mask Filter Replacement
2) Flow control: Flow control to 19.4 LPM, 0.6 LPM
3) Check flow rate with flow meter after connecting with CPC
4) After adjusting the flow rate, check leak test (blank)-> CPC 0 value by connecting with DMA.
-> sheath 6.0l pm / particle diameter 0nm-> sheath (6): CPC (0.6) = 10: 1
5) Change to DMA 50,100,200,300 nm to check down-stream kPa and CPC
6) Calculation formula for mask particle removal (dust collection) efficiency evaluation
* Mask particle removal (dust collection) efficiency efficiency (%) = (1-down / up) * 100
* pressure drop (kPa) = check down value

As a result of measuring the removal efficiency of mask particles (%) by measuring before and after 50-300 nm particle dust penetration amount in air in the general mask (HIP's) and the fine dust mask (KF84), the fine dust mask was 50-300. Particle removal (dust collection) efficiency is about 89-100% for nm-size fine dust particles, whereas general masks have particle removal efficiency of about 39-57% for 50-300 nm fine dust particles It was confirmed (Fig. 19).

 In addition, a 250 ppm solution of distilled water or a fatty acid-oligopeptide complex (distilled water dilution) is sprayed once on a general mask, and then the amount of fine dust particles having a size of 50 to 300 nm in the air is quantified before and after to remove mask particles (dust). As a result of measuring the collection efficiency (%), the collection collection efficiency was similar in the general mask without spraying anything and the distilled water spraying, and compared with the normal mask without spraying nothing, the fatty acid-peptide complex (Palmitic acid-KTTKS, In the mask treated with Palmitic acid-RRRRR, Palmitic acid-KKKKK, Capric acid-RRRRR, and Capric acid-RRRRRRRRR, the dust collection efficiency was increased. It was confirmed to be excellent (FIGS. 20 and 21).

<110> ANYGEN CO., LTD. <120> Cosmetic composition for eliminating or adsorbing particulate          matter comprising peptide complex as effective component <130> PN19194 <160> 30 <170> KoPatentIn 3.0 <210> 1 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 1 Ala Ala Pro Val   One <210> 2 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 2 Ala Ala Pro   One <210> 3 <211> 2 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 3 Ala Ala   One <210> 4 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 4 Gly his lys   One <210> 5 <211> 2 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 5 Gly his   One <210> 6 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 6 Lys Thr Thr Lys Ser   1 5 <210> 7 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 7 Lys Thr Thr Lys   One <210> 8 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 8 Lys Thr Thr   One <210> 9 <211> 2 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 9 Lys thro   One <210> 10 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 10 Glu Glu Met Gln Arg Arg   1 5 <210> 11 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 11 Glu Glu Met Gln Arg   1 5 <210> 12 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 12 Glu Glu Met Gln   One <210> 13 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 13 Glu glu met   One <210> 14 <211> 2 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 14 Glu glu   One <210> 15 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 15 Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys   1 5 <210> 16 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 16 Lys Lys Lys Lys Lys   1 5 <210> 17 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 17 Lys Lys Lys   One <210> 18 <211> 2 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 18 Lys Lys   One <210> 19 <211> 2 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 19 Gly pro   One <210> 20 <211> 2 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 20 Gly pro   One <210> 21 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 21 Gly Gln Pro Arg   One <210> 22 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 22 Gly gln pro   One <210> 23 <211> 2 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 23 Gly gln   One <210> 24 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 24 Thr Thr Lys Ser   One <210> 25 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 25 Thr Lys Ser   One <210> 26 <211> 2 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 26 Thr lys   One <210> 27 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 27 Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg   1 5 <210> 28 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 28 Arg Arg Arg Arg Arg   1 5 <210> 29 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 29 Arg Arg Arg   One <210> 30 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> peptide <400> 30 Leu Leu Trp Ile Ala Leu Arg Lys Lys   1 5

Claims (5)

amino acid; Or N terminal, C terminal or side chain of the oligopeptide; fine dust removal or adsorption cosmetic composition containing a complex in which the fatty acid is bonded as an active ingredient. According to claim 1, wherein the oligopeptide is a fine dust removal or adsorption cosmetic composition, characterized in that at least one selected from the group consisting of the amino acid sequence of SEQ ID NO: 1 to 30. According to claim 1, wherein the fatty acid is a fine dust removal or adsorption cosmetic composition, characterized in that the saturated fatty acid or unsaturated fatty acid. The method of claim 3, wherein the saturated fatty acid is caproic acid (Caproic acid, C6: 0), caprylic acid (Caprylic acid, C8: 0), Pelargonic acid (Pelargonic acid, C9: 0), capric acid (Capric acid, C10: 0), Undecylic acid (C11: 0), Lauric acid (C12: 0), Myristic acid (C14: 0), Pentadecylic acid (C15 : 0), Palmitic acid (C16: 0), Margaric acid (C17: 0), Stearic acid (Stearic acid, C18: 0), Nonadecylic acid (C19: 0) , Arachidic acid (C20: 0), Henicosylic acid (C21: 0), Behenic acid (C22: 0), Tricosylic acid (Crico: 20), league Lignoceric acid (C24: 0), pentacosylic acid (C25: 0), cerotic acid (C26: 0), heptacosylic acid (C27: 0), montanic acid ( Montanic acid, C28: 0), nonacosylic acid (C29: 0), melissic acid (C30: 0), henatriacontylic acid (C31: 0), lacse Acid (Lacceroic acid, C32: 0), Psyllic acid (C33: 0), Gedic acid (C34: 0), Ceroplastic acid (C35: 0), and hexatriacontylic acid ( Hexatriacontylic acid, C36: 0) at least one selected from the group consisting of fine dust removal or cosmetic composition for adsorption. The method of claim 3, wherein the unsaturated fatty acid is palmitoleic acid (Palmitoleic acid, C16: 1), oleic acid (Oleic acid, C18: 1), elideic acid (Elaidic acid, C18: 1), petroleic acid (Petroselinic acid) , C18: 1), Vaccenic acid (C18: 1), Gondoic acid (C20: 1), Erucic acid (C22: 1), Nervonic acid (C24: 1) , Linoleic acid (C18: 2), cis-Linolenic acid (C18: 3), Punicic acid (C18: 3), Eleostearic acid (C18: 3), Stearidonic acid (C18: 4), arachidonic acid (C20: 4), eicosapentaenoic acid (C20: 5), docosapentaenoic acid (Docosapentaenoic acid, C22: 5) , Fine dust, characterized in that at least one selected from the group consisting of adrenic acid (C22: 4), docosahexaenoic acid (Docosahexaenoic acid, C22: 6) and ricinoleic acid (C18: 1) Cosmetic composition for removal or adsorption.

Images