WO2013141576A1 - Method for stabilizing ramalin using porous matrix, and stablized ramalin solution - Google Patents

Method for stabilizing ramalin using porous matrix, and stablized ramalin solution Download PDF

Info

Publication number
WO2013141576A1
WO2013141576A1 PCT/KR2013/002247 KR2013002247W WO2013141576A1 WO 2013141576 A1 WO2013141576 A1 WO 2013141576A1 KR 2013002247 W KR2013002247 W KR 2013002247W WO 2013141576 A1 WO2013141576 A1 WO 2013141576A1
Authority
WO
WIPO (PCT)
Prior art keywords
ramalin
solution
porous matrix
hydrophilic
stabilizing
Prior art date
Application number
PCT/KR2013/002247
Other languages
French (fr)
Korean (ko)
Inventor
임정한
김일찬
김덕규
한세종
이형석
다타바타라이 하리
박수진
Original Assignee
한국해양연구원
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 한국해양연구원 filed Critical 한국해양연구원
Publication of WO2013141576A1 publication Critical patent/WO2013141576A1/en

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
    • A61K31/198—Alpha-amino acids, e.g. alanine or edetic acid [EDTA]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C241/00—Preparation of compounds containing chains of nitrogen atoms singly-bound to each other, e.g. hydrazines, triazanes
    • C07C241/04—Preparation of hydrazides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A61K31/135—Amines having aromatic rings, e.g. ketamine, nortriptyline
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
    • A61K8/0241—Containing particulates characterized by their shape and/or structure
    • A61K8/0283—Matrix particles
    • A61K8/0287—Matrix particles the particulate containing a solid-in-solid dispersion
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
    • A61K8/44—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof
    • A61K8/442—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof substituted by amido group(s)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/58—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing atoms other than carbon, hydrogen, halogen, oxygen, nitrogen, sulfur or phosphorus
    • A61K8/585—Organosilicon compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C243/00—Compounds containing chains of nitrogen atoms singly-bound to each other, e.g. hydrazines, triazanes
    • C07C243/10—Hydrazines
    • C07C243/22—Hydrazines having nitrogen atoms of hydrazine groups bound to carbon atoms of six-membered aromatic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/52—Stabilizers

Definitions

  • the present invention relates to a method for stabilizing ramalin and stabilized ramalin solution, and more particularly, after adsorbing ramalin to a hydrophilic porous matrix, modifying the surface of the porous matrix and dispersing it in an organic solvent.
  • a method of stabilizing marlin and thereby a stabilized ramalline solution is a method for stabilizing marlin and thereby a stabilized ramalline solution.
  • Lichens are a symbiotic association of mycobiant, microalga and / or cyanobacteria. Lichens contain a variety of secondary metabolites (Ahmadjin V., The lichen symbiosis, Wiley, New York , pp.1-6, 1993), but it is difficult to collect sufficient natural samples, and mass cultivation techniques are unknown. As a result, research is less than higher plants.
  • tissue culture method of lichens mass production method of metabolites and biochemical analysis method have been improved, researches on the metabolites they produce are being actively conducted (Behera, BC et al., Lebensm.Wiss.Technol. , 39: 805, 2006), fatty acids, depsides and depsidones, debenzofurans, diterpenes, anthraquinones, anthraquinones, which have several biological activities such as antibacterial, antibiotic and antioxidant ), Compounds including naphtoquinones, usninic acid, p ⁇ lvinic acids, xanthones and epidithiopiperazinediones have been isolated from lichens ( Muller, K., Appl. Microbiol.Biotechnol ., 56: 9-16, 2001).
  • Ramalina terebrata is a native lichen that colonizes King George Island in the Antarctic Peninsula and can be easily collected from various parts of King George Island.
  • the present inventors have isolated a novel compound called Ramalin (Ramalin) having excellent antioxidant activity while studying the Antarctic lichen Ramalina terebrata (Korean Patent Publication No. 10-2010-0052130).
  • Ramalin has been reported to have excellent anti-inflammatory activity (Korean Patent Application No. 10-2010-0052551).
  • the present inventors have made diligent efforts to develop a method for stabilizing ramallin that can maintain the titer of ramalin even when stored at a high temperature without long effect on temperature, and as a result of forming a porous matrix with a hydrophilic aerosol. After adsorbing the RAMALIN and modifying the surface and dispersing it in an organic solvent, it was confirmed that the high titer was maintained even during long-term storage and high titer even at high temperature, thereby completing the present invention.
  • An object of the present invention is to provide a method for stabilizing RAMALIN and thus stabilized RAMALIN solution, which can stabilize unstable RAMALIN even for long term storage.
  • the present invention provides a method for stabilizing Ramalin represented by the following formula (1) comprising the following steps:
  • the present invention also provides a stabilized ramalline solution, which is prepared by the above method.
  • the present invention also provides a cosmetic composition for whitening or improving wrinkles containing the stabilized ramine solution as an active ingredient.
  • 1 is a schematic view showing that the hydroxyl group of the hydrophilic airgel and ramalin formed the first physicochemical stabilization through hydrogen bonding.
  • FIG. 2 is a schematic showing that the hydrophilic airgel matrix is hydrophobically modified by lecithin.
  • Figure 3 is a schematic diagram showing the appearance of the o / w nano-emulsion particles surrounding the hydrophilic airgel particles adsorbed to the RAMALIN.
  • Figure 4 is a graph showing the titer of the stabilized ramalin solution according to the present invention at 25 °C and 45 °C storage for 4 weeks.
  • the present invention relates to a method for stabilizing ramalin represented by the following Chemical Formula 1 in one aspect, and more particularly, it includes the following steps:
  • Ramalin of the present invention is a compound having an antioxidant activity isolated from the Antarctic lichen, Ramalina terebrata, The ramalin is confirmed by using a high resolution ESI-MS, molecular weight is 254, the molecular formula C 11 H 16 N 3 O
  • the compound of 4 has a structure represented by the formula (1), because it is a compound separated from ramalina terebrata it was named as RAMALIN.
  • the porous matrix carrying the ramine is water-soluble, preferably the water-soluble porous matrix is composed of a hydrophilic airgel.
  • Ramalin is a water-soluble substance that is insoluble in a general organic solvent. In order to adsorb it into a porous matrix, first, an aqueous solution in which ramalin is dissolved must be prepared, followed by mixing and adsorbing the porous matrix in the mixture.
  • the porous matrix used is hydrophobically modified, it is difficult to disperse in the aqueous solution, and since the ramalline is particularly unstable in the aqueous solution, it should be placed in the oil phase rather than in the aqueous phase to prevent the hydrolysis reaction by the aqueous solution.
  • Hydrophilic airgel was prepared by preparing a wet gel using a sol-gel process and applying a supercritical fluid drying process thereto.
  • a general sol-gel process for preparing the wet gel is to obtain an alcohol by hydrolysis and polycondensation of silicon alkoxide, as follows.
  • silicon alkoxide various materials such as alumina alkoxide may be used as well as silicon.
  • hydrolysis proceeds rapidly in an acidic atmosphere (1)
  • condensation polymerization proceeds rapidly in a basic atmosphere (2).
  • Hydrophilic aerogels can be prepared by using water, alcohols and metal alkoxides as raw materials using acidic or basic catalysts.
  • tetra-alkoxysilane is used as the metal alkoxide.
  • the main parameters of the sol-gel process are the type of alkoxy silane solution, the H 2 O / Si ratio, and the pH of the reaction solution.
  • the pH change effect of the reaction solution can change the reaction rate of the hydrolysis and polycondensation reaction, it is also possible to control the reaction mechanism (reaction mechanism).
  • the hydrophilic aerogel is a mixture of tetramethylortho-silicate: water: methanol in a ratio of 1: 1.5: 1 under citric acid catalyst, and stirred in a mixture of NH, Tetramethyl-oso-silicate: water under 4 OH catalyst was prepared by adding a solution of isopropanol as a dilution solvent in a ratio of 1: 2.5 to form a gel, followed by removing the solvent and drying.
  • the hydrophilic airgel thus prepared is placed in an aqueous solution and sufficiently wetted to infiltrate water into the matrix. After slowly adding dropwise aqueous solution in which the RAMALIN is dissolved, the mixture is stirred to adsorb the RAMALIN inside the hydrophilic airgel to induce hydrogen bonding between the hydroxyl group (-OH) and the RAMALIN of the hydrophilic airgel and thereby primarily stabilize the physicochemical properties.
  • the form was prepared (FIG. 1).
  • the surface of the porous matrix adsorbed with the RAMALIN is then hydrophobically modified, characterized in that it is hydrophobically modified by treating the amphoteric surfactant or surfactant compound.
  • Amphiphilic compounds can be used as long as they can hydrophobically modify the surface of the hydrophilic porous matrix, for example lecithin, phosphatidyl ethanolamine, lyso lecithin, aminopropyl laurylglutamine, disodium tallow Disodium tallowam- phodiacetate, Cocoamphodipropionic acid, Disodium caproamphodiacetate, Cocamidopropyl betaine, Sodium C12-15 alkoxypropyl imide No propionate (Sodium C12-15 alkoxypropyl Iminodipropionate), Quaternium-85 (Quaternium-85) and the like can be used, but are not limited thereto.
  • lecithin was used, and after adsorbing ramalin inside a hydrophilic airgel matrix, it was placed in an ethanol mixture containing lecithin and distilled under reduced pressure to remove ethanol.
  • the hydrophilic group of the component phosphotidylcholine is oriented toward the surface of the airgel and the hydrophobic group of the phosphotidylcholine is oriented to the outside to facilitate dispersion in the oil phase (FIG. 2).
  • the hydrophobically modified porous matrix is dispersed in a hydrophobic organic solvent, wherein the organic solvent is caprylic / capric triglyceride, squalane, silicone oil, mineral oil, vegetable oil and mixtures thereof. Characterized in that it is selected from the group consisting of, but are not limited to these.
  • the hydrophobically modified ramal adsorbed porous matrix was treated with a mixture of glycerin and caprylic / capric triglycerides.
  • Glycerin is added to the mixture of caprylic / capric triglycerides, additionally dissolved by adding lecithin, purified water, and then passed through a microfludizer to make caprylic / capric triglycerides into the inner phase and glycerin to the trauma.
  • the o / w nanoemulsion particles are enclosed with the hydrophilic airgel particles adsorbed with ramallin to prepare a suspension in which the porous matrix particles having the submicron units are dispersed (FIG. 3).
  • the stabilized ramalin solution prepared by the method of the present invention showed a high titer retention of about 96% even after 4 weeks in storage at 25 ° C, and a 61% titer retention rate at 45 ° C. Therefore, the method of stabilizing the RAMALIN has been shown to greatly aid in improving the stability of RAMALIN. That is, the method according to the present invention can stabilize the RAMALIN to maintain a nearly similar titer in comparison with the RAMALIN standard even during long-term storage, so that the RAMALIN having excellent antioxidant and anti-inflammatory effects can be stored for a long time. It is useful in keeping the titer at a high level even at the same high temperature for a long time to facilitate the storage of ramalin.
  • the present invention relates to a stabilized ramalline solution, which is prepared by the above method.
  • the cosmetic composition containing the stabilized ramalin solution prepared by the method has an effect of whitening and wrinkle improvement. Accordingly, the present invention relates to a cosmetic composition containing the stabilized ramine solution as an active ingredient in another aspect.
  • the stabilized RAMALIN solution according to the present invention may be preferably characterized in that it comprises 0.0001% to 10% based on the total weight of the cosmetic composition.
  • the cosmetic composition of the present invention can be prepared in the form of general emulsion formulations and solubilized formulations.
  • Cosmetics of the emulsified formulations include nutrient cosmetics, creams, essences, etc., and cosmetics of the solubilized formulations are flexible cosmetics.
  • the cosmetic composition according to the present invention may prepare a formulation such as a lotion, an emulsion, a cream, an essence, a cosmetic ointment, a spray, a gel, a pack, a sunscreen, a makeup base, a foundation, a powder, a makeup cleaner and a cleaner
  • Suitable cosmetic formulations include, for example, emulsions, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes), nonionic vesicles obtained by dispersing an oil phase in a solution, gel, solid or pasty anhydrous product, aqueous phase, for example. It may be provided in the form of a dispersant, cream, skin, lotion, powder, ointment, spray or cone stick. It may also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.
  • Cosmetics of the present invention may further comprise fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionics.
  • fatty substances organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionics.
  • TMOS Tetramethylortho- silicate
  • citric acid C 6 H 8 O 7
  • NH 4 OH NH 4 OH
  • citric acid as a catalyst
  • the pH is maintained at 2.0 and stirred for 1 hour at room temperature / atmospheric pressure.
  • the methanol was removed using a vacuum evaporator.
  • the whole solution prepared in the second step was put together in the first step solution and mixed for 2 hours at room temperature / atmospheric pressure.
  • the raw material solutions of the second stage should not be added individually and sequentially to the solution prepared in the first stage.
  • the sol solution prepared above was poured into a sealed container, sealed, and placed in a 50 ° C. thermostatic bath to induce gelation. Generally, the gel is formed within 3 hours under the above conditions.
  • the wet gel prepared in the sol-gel process was placed in a supercritical reactor and the reactor was sealed. Ethanol was supplied to the top of the supercritical reactor using a high pressure liquid pump to pressurize the pressure to 50 atm at room temperature. When 50 atm is reached, ethanol is fed to the top of the supercritical reactor, and the liquid mixture is discharged to the bottom and maintained at 50 atm (using gravity direction). This is the catalyst and unreacted water (H 2 O) remaining in the wet gel. It is to remove. In this case, the total amount of ethanol supplied is about twice the internal volume of the supercritical reactor or about three times the volume of the wet gel, and a flow rate of 5 mL / min is sufficient.
  • liquid carbon dioxide was fed to the top of the reactor and ethanol was released to the bottom.
  • the temperature at this time was 30 degrees C or less, and the pressure necessarily maintained 70 or more atmospheres.
  • the valves of the upper and lower parts of the reactor are closed and the heater of the reactor is operated to raise the temperature inside the reactor to 40 ° C., at which time the pressure is increased as the temperature increases.
  • the pressure was maintained at 80 to 100 atm, and when the pressure rises above 10 atm, the valve at the bottom of the reactor is opened to adjust the pressure.
  • the temperature inside the reactor is maintained at 40 ° C, and carbon dioxide is released using a rear pressure regulator, and the carbon dioxide discharge flow rate is about 2 atm / min (0.2 MPa / min).
  • the reactor was opened without cooling to obtain a dried hydrophilic airgel.
  • the recovered hydrophilic airgel was ground to a powder of desired size.
  • Ramalina terebrata is a native lichen that colonizes King George Island in Antarctica and can be easily collected from all over King George Island.
  • the solvent system used was 10 minutes for a 0% methanol-water mixed solution for water mixed with 0.1% formic acid, 20 minutes for a 20% methanol-water mixed solution and 30 minutes for a 100% methanol solution.
  • the flow rate was 2 mL / min.
  • the compound was absorbed at 280 nm ultraviolet.
  • Example 1-4 After dissolving 3.1 g of lecithin in 50 g of ethanol heated to 50 ° C., 31 g of the ramalin-hydrophilic aerogel obtained in Example 1-4 was slowly added, dispersed evenly using an azimixer, and then distilled under reduced pressure at 60 ° C. to give a surface treatment. 32 g of hydrophilic airgel powder was obtained.
  • Example 1-5 28 g of the powder obtained in Example 1-5 was added to 210 g of caprylic / capric triglyceride, and 357 g of glycerin was added while warming to 60 ° C., and warmed to 70 ° C., and 35 g of lecithin was added thereto to completely dissolve it. After adding 70 g of purified water to the mixture and stirring at 4,000 rpm for 20 minutes, the mixture was passed three times at 1,000 bar using a high pressure homogenizer (microfludizer).
  • a high pressure homogenizer microfludizer
  • the concentration was calculated by comparing the area values as shown below. The results were compared at 100%.
  • the stabilizing ramalin solution was prepared at the same concentration as the stabilizing ramalin solution in the incubator at 25 °C and 45 °C observed for four weeks at intervals of one week, stabilizing ramalin solution was maintained at a high titer of about 96% even after 4 weeks when stored at 25 °C, while unstable aqueous solution of unstable ramaline lost more than 50% of the titer in one week.
  • the stabilized ramalin solution showed a titer retention rate of 61% at 45 ° C., and the unstabilized ramalin aqueous solution destroyed more than 90% of the total titer at 45 ° C. in one week. Therefore, the method of stabilizing the RAMALIN has been shown to greatly aid in improving the stability of RAMALIN.
  • Example 1 After preparing a cosmetic formulation comprising the stabilized ramallin prepared in Example 1 as shown in Table 1 below, divided 30 groups of 20 to 60-year-old female subjects into two groups daily cosmetic and control cosmetics containing ramalin Approximately 1 g was applied to the face for 4 weeks in the morning and evening, and then the subject's facial skin brightness and wrinkle improvement were evaluated every week.
  • the degree of improvement of skin brightness was measured using ARAMO TS (Aram Huvis, South Korea), and the change in the measured value is shown in Table 2 below.
  • the wrinkle improvement effect was obtained by taking a replica under the subject's eyes (Xantopren, Bayer) before the experiment began, and after 4 weeks after the experiment was completed, a replica was taken from the same area under the eyes to analyze the wrinkles.
  • Wrinkle density was measured by two-dimensional analysis, and the results of the measurement of wrinkle density by image analysis are shown in Table 3 by averaging the reduction ratio of the wrinkle density before use.
  • the stabilization method of ramalin according to the present invention is capable of stabilizing ramalin so as to maintain a nearly similar titer compared to the ramalin standard even in long-term storage, which has the advantage of allowing the long-term storage of ramin excellent in antioxidant and anti-inflammatory effects. have.
  • the method according to the present invention is useful in that it can be maintained at a high level without rapidly lowering the titer even when stored at a high temperature such as 45 ° C. to facilitate the storage of ramalin.
  • the stabilized RAMALIN solution according to the present invention can be used as functional cosmetics for whitening and wrinkle improvement as it has an effect of improving the brightness of the skin and improving wrinkles.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Organic Chemistry (AREA)
  • Epidemiology (AREA)
  • Birds (AREA)
  • Dermatology (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Dispersion Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Mathematical Physics (AREA)
  • Gerontology & Geriatric Medicine (AREA)
  • Cosmetics (AREA)
  • Medicinal Preparation (AREA)

Abstract

The present invention relates to a method for stabilizing ramalin and a stabilized ramalin solution, and more specifically to a method for stabilizing ramalin and a solution of thus stabilized ramalin, the method for stabilizing ramalin being characterized by adhering ramalin to a hydrophilic and porous matrix, then modifying the surface thereof so as to disperse same in an organic solvent. A method for stabilizing ramalin according to the present invention allows stabilizing ramalin in such a way as to maintain similar potency in comparison to the standard preparation of ramalin even when stored for an extended period of time, thus having the benefit of allowing an extended storage of ramalin, which has excellent antioxidant and anti-inflammatory activities. Furthermore, the method according the present invention facilitates storage of ramalin as the potency thereof is maintained even when stored for an extended period at a high temperature such as 45 °C.

Description

다공성 매트릭스를 이용한 라말린의 안정화 방법 및 안정화된 라말린 용액Stabilization of Ramalin Using Porous Matrix and Stabilized Ramalin Solution
본 발명은 라말린의 안정화 방법 및 안정화된 라말린 용액에 관한 것으로, 보다 상세하게는 친수성의 다공성 매트릭스에 라말린을 흡착시킨 후 다공성 매트릭스의 표면을 개질하여 유기용매에 분산시키는 것을 특징으로 하는 라말린의 안정화 방법 및 이에 의한 안정화된 라말린 용액에 관한 것이다.The present invention relates to a method for stabilizing ramalin and stabilized ramalin solution, and more particularly, after adsorbing ramalin to a hydrophilic porous matrix, modifying the surface of the porous matrix and dispersing it in an organic solvent. A method of stabilizing marlin and thereby a stabilized ramalline solution.
지의류는 곰팡이(mycobiant), 미세조류 (microalga) 및/또는 남조류 (cyanobacteria)의 공생연합이다. 지의류는 다양한 이차대사산물을 함유하나(Ahmadjin V., The lichen symbiosis, Wiley, New York, pp.1-6, 1993), 충분한 양의 천연시료를 수집하기가 어렵고, 대량재배 기술이 알려져 있지 않기 때문에, 고등식물보다는 연구가 미진하였다. Lichens are a symbiotic association of mycobiant, microalga and / or cyanobacteria. Lichens contain a variety of secondary metabolites (Ahmadjin V., The lichen symbiosis, Wiley, New York , pp.1-6, 1993), but it is difficult to collect sufficient natural samples, and mass cultivation techniques are unknown. As a result, research is less than higher plants.
현재 지의류의 조직배양법, 대사체 대량생산 방법 및 생화학적 분석 방법 등이 개선됨에 따라, 이들이 생산하는 대사체에 대한 연구가 활발히 진행되고 있고(Behera, B.C. et al., Lebensm. Wiss. Technol., 39:805, 2006), 항균, 항생 및 항산화 등의 여러 생물학적 활성을 가지는 지방산, 뎁시드 (depside) 및 뎁시돈 (depsidones), 디벤조푸란 (debenzofurans), 디터펜 (diterpenes), 안트라퀴논 (anthraquinones), 나프토퀴논 (naphtoquinones), 우스닉산 (usninic acid), 풀비닉 산 (pμlvinic acids), 잔톤 (xanthones) 및 에피디티오피퍼라진이온 (epidithiopiperazinediones)을 포함하는 화합물들이 지의류로부터 분리된 바 있다(Muller, K., Appl. Microbiol. Biotechnol., 56:9-16, 2001). As the tissue culture method of lichens, mass production method of metabolites and biochemical analysis method have been improved, researches on the metabolites they produce are being actively conducted (Behera, BC et al., Lebensm.Wiss.Technol. , 39: 805, 2006), fatty acids, depsides and depsidones, debenzofurans, diterpenes, anthraquinones, anthraquinones, which have several biological activities such as antibacterial, antibiotic and antioxidant ), Compounds including naphtoquinones, usninic acid, pμlvinic acids, xanthones and epidithiopiperazinediones have been isolated from lichens ( Muller, K., Appl. Microbiol.Biotechnol ., 56: 9-16, 2001).
라말리나 테레브라타 (Ramalina terebrata)는 남극반도 킹조지섬 등에 군락을 이루어 자생하는 지의류로서, 킹조지섬의 곳곳에서 용이하게 채취할 수 있다. 본 발명자들은 상기 남극 지의류 Ramalina terebrata를 연구하던 중에 우수한 항산화 활성을 가진 라말린(Ramalin)이라는 신규 화합물을 분리한 바 있다(대한민국 공개특허공보 제10-2010-0052130호). 또한, 라말린은 우수한 항염증 활성을 가진 것이 보고된 바 있다(대한민국 특허출원번호 제10-2010-0052551호). Ramalina terebrata is a native lichen that colonizes King George Island in the Antarctic Peninsula and can be easily collected from various parts of King George Island. The present inventors have isolated a novel compound called Ramalin (Ramalin) having excellent antioxidant activity while studying the Antarctic lichen Ramalina terebrata (Korean Patent Publication No. 10-2010-0052130). In addition, ramalin has been reported to have excellent anti-inflammatory activity (Korean Patent Application No. 10-2010-0052551).
라말린을 상업적으로 대량 생산하여 판매하기 위해서는 장기간 보관시에도 그 역가가 동일 또는 유사하게 유지되는 것이 필요하나, 라말린은 매우 불안정한 성질을 갖기 때문에 이의 안정화 방법이 요구되었다. In order to commercially produce and sell ramalin, it is necessary that the titer remains the same or similar even after long-term storage, but since stabilization is very unstable, a method of stabilization thereof has been required.
이에 본 발명자들은 장기간 보관시에도, 아울러 온도에 큰 영향을 받지 않고 고온에서 보관할 때도 라말린의 역가를 유지할 수 있는 라말린의 안정화 방법을 개발하기 위하여 예의 노력한 결과, 친수성 에어로졸로 다공성 매트릭스를 형성시킨 후 이에 라말린을 흡착시키고 표면을 개질하여 유기용매에 분산시키는 경우 장기간 보관시에도 높은 역가를 유지하고 고온에서도 높은 역가를 보임을 확인하고, 본 발명을 완성하게 되었다.Accordingly, the present inventors have made diligent efforts to develop a method for stabilizing ramallin that can maintain the titer of ramalin even when stored at a high temperature without long effect on temperature, and as a result of forming a porous matrix with a hydrophilic aerosol. After adsorbing the RAMALIN and modifying the surface and dispersing it in an organic solvent, it was confirmed that the high titer was maintained even during long-term storage and high titer even at high temperature, thereby completing the present invention.
발명의 요약Summary of the Invention
본 발명의 목적은 불안정한 라말린을 장기간 보관시에도 안정화시킬 수 있는 라말린의 안정화 방법 및 이에 따른 안정화된 라말린 용액을 제공하는 데 있다.SUMMARY OF THE INVENTION An object of the present invention is to provide a method for stabilizing RAMALIN and thus stabilized RAMALIN solution, which can stabilize unstable RAMALIN even for long term storage.
상기 목적을 달성하기 위하여, 본 발명은 다음 단계를 포함하는 하기 화학식 1로 표시되는 라말린의 안정화 방법을 제공한다:In order to achieve the above object, the present invention provides a method for stabilizing Ramalin represented by the following formula (1) comprising the following steps:
(a) 친수성의 다공성 매트릭스에 라말린을 흡착시키는 단계;(a) adsorbing ramalin to a hydrophilic porous matrix;
(b) 상기 라말린이 흡착된 다공성 매트릭스의 표면을 소수성으로 개질하는 단계; 및(b) hydrophobically modifying the surface of said ramal adsorbed porous matrix; And
(c) 상기 소수성으로 개질된 다공성 매트릭스를 유기 용매에 분산시키는 단계(c) dispersing the hydrophobically modified porous matrix in an organic solvent
화학식 1
Figure PCTKR2013002247-appb-C000001
 Formula 1
Figure PCTKR2013002247-appb-C000001
본 발명은 또한, 상기 방법에 의하여 제조되는 것을 특징으로 하는 안정화된 라말린 용액을 제공한다. The present invention also provides a stabilized ramalline solution, which is prepared by the above method.
본 발명은 또한, 상기 안정화된 라말린 용액을 유효성분으로 함유하는 미백 또는 주름 개선용 화장료 조성물을 제공한다. The present invention also provides a cosmetic composition for whitening or improving wrinkles containing the stabilized ramine solution as an active ingredient.
도 1은 친수성 에어로겔의 수산화기와 라말린이 수소결합을 통하여 1차적인 물리화학적 안정화를 형성한 것을 보여주는 개략도이다. 1 is a schematic view showing that the hydroxyl group of the hydrophilic airgel and ramalin formed the first physicochemical stabilization through hydrogen bonding.
도 2는 친수성 에어로겔 매트릭스가 레시틴에 의하여 소수성으로 개질되는 것을 보이는 개략도이다. 2 is a schematic showing that the hydrophilic airgel matrix is hydrophobically modified by lecithin.
도 3은 o/w 나노 에멀젼 입자가 라말린이 흡착된 친수성 에어로겔 입자를 둘러싼 모습을 보이는 개략도이다. Figure 3 is a schematic diagram showing the appearance of the o / w nano-emulsion particles surrounding the hydrophilic airgel particles adsorbed to the RAMALIN.
도 4는 본 발명에 따른 안정화된 라말린 용액이 4주 동안 25℃ 및 45℃ 보관 시 역가를 나타내는 그래프이다. Figure 4 is a graph showing the titer of the stabilized ramalin solution according to the present invention at 25 ℃ and 45 ℃ storage for 4 weeks.
발명의 상세한 설명 및 바람직한 구현예Detailed Description of the Invention and Preferred Embodiments
다른 식으로 정의되지 않는 한, 본 명세서에서 사용된 모든 기술적 및 과학적 용어들은 본 발명이 속하는 기술분야에서 숙련된 전문가에 의해서 통상적으로 이해되는 것과 동일한 의미를 갖는다. 일반적으로, 본 명세서에서 사용된 명명법은 본 기술분야에서 잘 알려져 있고 통상적으로 사용되는 것이다.Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is well known and commonly used in the art.
본 발명은 일 관점에서, 하기 화학식 1로 표시되는 라말린의 안정화 방법에 관한 것으로, 보다 상세하게는 다음 단계를 포함하는 것을 특징으로 한다:The present invention relates to a method for stabilizing ramalin represented by the following Chemical Formula 1 in one aspect, and more particularly, it includes the following steps:
(a) 친수성의 다공성 매트릭스에 라말린을 흡착시키는 단계;(a) adsorbing ramalin to a hydrophilic porous matrix;
(b) 상기 라말린이 흡착된 다공성 매트릭스의 표면을 소수성으로 개질하는 단계; 및(b) hydrophobically modifying the surface of said ramal adsorbed porous matrix; And
(c) 상기 소수성으로 개질된 다공성 매트릭스를 유기 용매에 분산시키는 단계(c) dispersing the hydrophobically modified porous matrix in an organic solvent
화학식 1 Formula 1
Figure PCTKR2013002247-appb-I000001
Figure PCTKR2013002247-appb-I000001
본 발명의 라말린은 남극 지의류인 라말리나 테레브라타로부터 분리한 항산화 활성을 갖는 화합물로, 상기 라말린은 고분해능 ESI-MS를 이용해 확인한 결과, 분자량이 254이고, 분자식 C11H16N3O4의 화합물로 화학식 1로 표시되는 구조를 가지며, 라말리나 테레브라타로부터 분리된 화합물이므로 라말린으로 명명되었다.Ramalin of the present invention is a compound having an antioxidant activity isolated from the Antarctic lichen, Ramalina terebrata, The ramalin is confirmed by using a high resolution ESI-MS, molecular weight is 254, the molecular formula C 11 H 16 N 3 O The compound of 4 has a structure represented by the formula (1), because it is a compound separated from ramalina terebrata it was named as RAMALIN.
본 발명에서는 라말린 활용을 위하여 고온에서 장기간 안정성이 요구되는 보존방법을 개발하기 위하여 노력한 결과, 다공성 매트릭스를 이용하는 매우 간단한 공정으로 라말린을 안정화할 수 있음을 확인하였다. In the present invention, as a result of the effort to develop a preservation method that requires long-term stability at high temperature for the use of ramalline, it was confirmed that it can be stabilized by a very simple process using a porous matrix.
본 발명에서 라말린을 담지하는 다공성 매트릭스는 수용성인 것을 특징으로 하는데, 바람직하게는 상기 수용성의 다공성 매트릭스는 친수성 에어로겔로 구성되는 것을 특징으로 한다. 라말린은 일반적인 유기용매에 녹지 않는 수용성 물질로 이를 다공성 매트릭스 내에 흡착을 하기 위해서는 먼저 라말린이 용해된 수용액을 제조한 후 이 혼합물 내에 다공성 매트릭스를 혼산 및 흡착 하여야 한다. 이때 사용되는 다공성 매트릭스가 소수성으로 개질이 되어 있다면 수용액 내에 분산하기 어렵고, 라말린은 특히 수용액내에서 매우 불안정하므로 이를 수상이 아니 유상에 위치 시켜야 수용액에 의한 가수분해 반응을 막을 수 있다. In the present invention, it is characterized in that the porous matrix carrying the ramine is water-soluble, preferably the water-soluble porous matrix is composed of a hydrophilic airgel. Ramalin is a water-soluble substance that is insoluble in a general organic solvent. In order to adsorb it into a porous matrix, first, an aqueous solution in which ramalin is dissolved must be prepared, followed by mixing and adsorbing the porous matrix in the mixture. At this time, if the porous matrix used is hydrophobically modified, it is difficult to disperse in the aqueous solution, and since the ramalline is particularly unstable in the aqueous solution, it should be placed in the oil phase rather than in the aqueous phase to prevent the hydrolysis reaction by the aqueous solution.
친수성 에어로겔은 졸-겔 (sol-gel) 공정을 이용하여 습윤겔(wet gel)을 제조하고 여기에 초임계 유체 건조공정을 적용하여 제조하였다. Hydrophilic airgel was prepared by preparing a wet gel using a sol-gel process and applying a supercritical fluid drying process thereto.
습윤겔을 제조하는 일반적인 졸-겔 공정은 다음과 같이 실리콘 알콕사이드(silicon alkoxide)의 가수분해와 축중합반응을 거쳐서 알코겔(alcogel)을 얻어내는 것이다. 원료로는 실리콘만이 아니라 알루미나 알콕사이드 등 다양한 물질을 사용할 수 있다. 이때 산성(acidic) 분위기에서는 가수분해가 빠르게 진행 되며(1), 반대로 염기성(basic) 분위기 하에서는 축중합반응이 빠르게 진행된다(2).A general sol-gel process for preparing the wet gel is to obtain an alcohol by hydrolysis and polycondensation of silicon alkoxide, as follows. As a raw material, various materials such as alumina alkoxide may be used as well as silicon. At this time, hydrolysis proceeds rapidly in an acidic atmosphere (1), whereas condensation polymerization proceeds rapidly in a basic atmosphere (2).
Figure PCTKR2013002247-appb-I000002
Figure PCTKR2013002247-appb-I000002
친수성 에어로겔은 산성 또는 염기성 촉매를 이용하여, 물, 알콜 및 금속 알콕사이드를 원료로 하여 제조할 수 있는데, 본 발명의 일 실시예에서는 금속 알콕사이드로 tetra-alkoxysilane을 사용하였다. Hydrophilic aerogels can be prepared by using water, alcohols and metal alkoxides as raw materials using acidic or basic catalysts. In one embodiment of the present invention, tetra-alkoxysilane is used as the metal alkoxide.
졸-겔 공정의 주요변수로는 alkoxy silane 용액의 종류, H2O/Si 비율, 그리고 반응용액의 pH 등 이다. 특히 반응용액의 pH 변화효과는 가수분해와 축중합반응의 반응속도를 변화시킬 수 있으며 반응기구(reaction mechanism)의 조절도 가능하다. The main parameters of the sol-gel process are the type of alkoxy silane solution, the H 2 O / Si ratio, and the pH of the reaction solution. In particular, the pH change effect of the reaction solution can change the reaction rate of the hydrolysis and polycondensation reaction, it is also possible to control the reaction mechanism (reaction mechanism).
본 발명에 있어서, 바람직하게는 상기 친수성 에어로겔은 시트르산 (citric acid) 촉매 하에서 테트라메틸-오소-실리케이트 (tetramethylortho-silicate): 물: 메탄올을 1: 1.5:1의 비로 혼합하여 교반한 혼합물에, NH4OH 촉매 하에서 테트라메틸-오소-실리케이트:물을 이소프로판올을 희석용매로 하여 1: 2.5의 비로 혼합한 용액을 가하여 겔을 형성한 다음, 용매를 제거하고 건조하여 제조된 것임을 특징으로 한다.In the present invention, preferably, the hydrophilic aerogel is a mixture of tetramethylortho-silicate: water: methanol in a ratio of 1: 1.5: 1 under citric acid catalyst, and stirred in a mixture of NH, Tetramethyl-oso-silicate: water under 4 OH catalyst was prepared by adding a solution of isopropanol as a dilution solvent in a ratio of 1: 2.5 to form a gel, followed by removing the solvent and drying.
이렇게 제조된 친수성 에어로겔을 수용액 내에 넣고 충분히 습윤(wetting) 시켜 매트릭스의 내부에 물을 침투시킨다. 이후 라말린이 용해 된 수용액을 천천히 적가 하면서 교반을 하여 친수성 에어로겔의 내부에 라말린을 흡착 시켜 친수성 에어로겔의 수산화기(-OH)와 라말린의 수소결합을 유도하고 이를 통하여 1차적으로 물리화학적인 안정화 형태를 제조하였다 (도 1). The hydrophilic airgel thus prepared is placed in an aqueous solution and sufficiently wetted to infiltrate water into the matrix. After slowly adding dropwise aqueous solution in which the RAMALIN is dissolved, the mixture is stirred to adsorb the RAMALIN inside the hydrophilic airgel to induce hydrogen bonding between the hydroxyl group (-OH) and the RAMALIN of the hydrophilic airgel and thereby primarily stabilize the physicochemical properties. The form was prepared (FIG. 1).
그 후, 라말린이 흡착된 다공성 매트릭스의 표면은 소수성으로 개질되는데, 이는 양쪽성 계면활성제 또는 계면활성제 화합물을 처리하여 소수성으로 개질하는 것을 특징으로 한다. 양친매성 화합물로는 친수성의 다공성 매트릭스 표면을 소수성으로 개질할 수 있는 것이면 어떠한 것이든 사용가능하며, 예시적으로 레시틴, 포스파티딜 에탄올아민, 리소레시틴, 아미노프로필 라우릴글루타민(aminopropyl laurylglutamine), 다이소듐 탈로우암포다이아세테이트 (Disodium tallowam- phodiacetate), 코코암포다이프로피오닉산(Cocoamphodipropionic acid), 다이소듐 카프로암포다이아세테이트(Disodium Caproamphodiacetate), 코카미도프로필 베타인(Cocamidopropyl Betain), 소듐 C12-15 알콕시프로필 이미노프로피오네이트(Sodium C12-15 alkoxypropyl Iminodipropionate), 쿼터니움-85(Quaternium-85) 등을 사용할 수 있으나 이들로 제한되는 것은 아니다. The surface of the porous matrix adsorbed with the RAMALIN is then hydrophobically modified, characterized in that it is hydrophobically modified by treating the amphoteric surfactant or surfactant compound. Amphiphilic compounds can be used as long as they can hydrophobically modify the surface of the hydrophilic porous matrix, for example lecithin, phosphatidyl ethanolamine, lyso lecithin, aminopropyl laurylglutamine, disodium tallow Disodium tallowam- phodiacetate, Cocoamphodipropionic acid, Disodium caproamphodiacetate, Cocamidopropyl betaine, Sodium C12-15 alkoxypropyl imide No propionate (Sodium C12-15 alkoxypropyl Iminodipropionate), Quaternium-85 (Quaternium-85) and the like can be used, but are not limited thereto.
본 발명의 실시예에서는 레시틴을 사용하였는데, 친수성 에어로겔 매트릭스 내부에 라말린을 흡착 후 이를 레시틴이 용해 된 에탄올 혼합물에 넣은 뒤 감압증류 하여 에탄올을 제거 하면 라말린이 흡착 된 친수성 에어로겔 표면에 레시틴의 주요성분인 포스포티딜콜린(phosphotidylcholine)의 친수성기가 에어로겔의 표면쪽으로 배향을 하고 포스포티딜콜린의 소수성기가 외부로 배향을 하게 되어 오일상에서 분산이 용이해 진다(도 2). In an embodiment of the present invention, lecithin was used, and after adsorbing ramalin inside a hydrophilic airgel matrix, it was placed in an ethanol mixture containing lecithin and distilled under reduced pressure to remove ethanol. The hydrophilic group of the component phosphotidylcholine is oriented toward the surface of the airgel and the hydrophobic group of the phosphotidylcholine is oriented to the outside to facilitate dispersion in the oil phase (FIG. 2).
본 발명의 방법에서, 상기 소수성으로 개질된 다공성 매트릭스를 소수성 유기 용매에 분산시키는데, 이때 유기 용매로는 카프릴릭/카프릭트리글리세라이드, 스쿠알란, 실리콘 오일, 미네랄 오일, 식물성 오일 및 이들의 혼합물로 구성된 군에서 선택되는 것을 특징으로 할 수 있으나 이들로 한정되는 것은 아니다. In the process of the invention, the hydrophobically modified porous matrix is dispersed in a hydrophobic organic solvent, wherein the organic solvent is caprylic / capric triglyceride, squalane, silicone oil, mineral oil, vegetable oil and mixtures thereof. Characterized in that it is selected from the group consisting of, but are not limited to these.
본 발명의 일 실시예에서는, 소수성으로 개질된 라말린을 흡착한 다공성 매트릭스를 글리세린과 카프릴릭/카프릭트리글리세라이드의 혼합물로 처리하였다. 글리세린을 카프릴릭/카프릭트리글리세라이드의 혼합물에 넣고 추가적으로 레시친을 추가하여 용해하고 정제수를 넣은 뒤 고압균질기(microfludizer)를 통과 시키면 추가적으로 카프릴릭/카프릭트리글리세라이드를 내상으로 하고 글리세린을 외상으로 하는 o/w 나노 에멀젼 입자가 라말린이 흡착 된 친수성 에어로겔 입자를 둘러 쌓게 되어 서브마이크론 단위를 갖는 다공성 매트릭스 입자가 분산된 현탁액을 제조하게 된다(도 3).In one embodiment of the present invention, the hydrophobically modified ramal adsorbed porous matrix was treated with a mixture of glycerin and caprylic / capric triglycerides. Glycerin is added to the mixture of caprylic / capric triglycerides, additionally dissolved by adding lecithin, purified water, and then passed through a microfludizer to make caprylic / capric triglycerides into the inner phase and glycerin to the trauma. The o / w nanoemulsion particles are enclosed with the hydrophilic airgel particles adsorbed with ramallin to prepare a suspension in which the porous matrix particles having the submicron units are dispersed (FIG. 3).
본 발명의 방법에 의하여 제조된 안정화된 라말린 용액은 25℃에서 보관 시 4주 후에도 약 96%의 높은 역가 유지율을 보여주었으며, 45℃에서는 61%의 역가 유지율을 보여주었다. 따라서 상기 라말린 안정화 방법은 라말린의 안정성 향상에 크게 도움을 주는 것으로 나타났다. 즉, 본 발명에 따른 방법은 장기간 보관시에도 라말린 표준품에 대비하여 거의 유사한 역가를 유지하도록 라말린을 안정화시킬 수 있어 항산화 및 항염증 효과가 뛰어난 라말린을 장기간 보관할 수 있게 하며, 45℃와 같은 고온에서도 장기간 보관 시 역가를 높은 수준으로 유지시킬 수 있어 라말린의 보관을 용이하게 하는 점에서 유용하다. 따라서, 본 발명은 다른 관점에서, 상기 방법에 의하여 제조되는 것을 특징으로 하는 안정화된 라말린 용액에 관한 것이다. The stabilized ramalin solution prepared by the method of the present invention showed a high titer retention of about 96% even after 4 weeks in storage at 25 ° C, and a 61% titer retention rate at 45 ° C. Therefore, the method of stabilizing the RAMALIN has been shown to greatly aid in improving the stability of RAMALIN. That is, the method according to the present invention can stabilize the RAMALIN to maintain a nearly similar titer in comparison with the RAMALIN standard even during long-term storage, so that the RAMALIN having excellent antioxidant and anti-inflammatory effects can be stored for a long time. It is useful in keeping the titer at a high level even at the same high temperature for a long time to facilitate the storage of ramalin. Thus, in another aspect, the present invention relates to a stabilized ramalline solution, which is prepared by the above method.
본 발명의 다른 실시예에서는 상기 방법에 의하여 제조된 안정화된 라말린 용액을 함유하는 화장료 조성물이 미백 및 주름 개선 효과가 있음을 확인하였다. 이에 본 발명은, 또 다른 관점에서 상기 안정화된 라말린 용액을 유효성분으로 함유하는 화장료 조성물에 관한 것이다.In another embodiment of the present invention it was confirmed that the cosmetic composition containing the stabilized ramalin solution prepared by the method has an effect of whitening and wrinkle improvement. Accordingly, the present invention relates to a cosmetic composition containing the stabilized ramine solution as an active ingredient in another aspect.
이때, 본 발명에 따른 안정화된 라말린 용액은 바람직하게는 화장료 조성물 총 중량에 대하여 0.0001% 내지 10%로 포함되는 것을 특징으로 할 수 있다. At this time, the stabilized RAMALIN solution according to the present invention may be preferably characterized in that it comprises 0.0001% to 10% based on the total weight of the cosmetic composition.
본 발명의 화장료 조성물은 일반적인 유화 제형 및 가용화 제형의 형태로 제조할 수 있다. 유화 제형의 화장품으로는 영양화장수, 크림, 에센스 등이 있으며, 가용화 제형의 화장품으로는 유연화장수가 있다. The cosmetic composition of the present invention can be prepared in the form of general emulsion formulations and solubilized formulations. Cosmetics of the emulsified formulations include nutrient cosmetics, creams, essences, etc., and cosmetics of the solubilized formulations are flexible cosmetics.
본 발명에 따른 화장료 조성물은 화장수, 유액, 크림, 에센스, 화장연고, 스프레이, 젤, 팩, 선 스크린, 메이크업 베이스, 파운데이션, 파우더, 메이크업 세정제 및 세정제 등의 제형을 제조될 수 있는데, 본 발명에 적합한 화장품의 제형으로는 예를 들면 용액, 겔, 고체 또는 반죽 무수 생성물, 수상에 유상을 분산시켜 얻은 에멀젼, 현탁액, 마이크로에멀젼, 마이크로캡슐, 미세과립구 또는 이온형(리포좀), 비이온형의 소낭 분산제의 형태, 크림, 스킨, 로션, 파우더, 연고, 스프레이 또는 콘실 스틱의 형태로 제공될 수 있다. 또한, 포말(foam)의 형태 또는 압축된 추진제를 더 함유한 에어로졸 조성물의 형태로도 제조될 수 있다. The cosmetic composition according to the present invention may prepare a formulation such as a lotion, an emulsion, a cream, an essence, a cosmetic ointment, a spray, a gel, a pack, a sunscreen, a makeup base, a foundation, a powder, a makeup cleaner and a cleaner, Suitable cosmetic formulations include, for example, emulsions, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes), nonionic vesicles obtained by dispersing an oil phase in a solution, gel, solid or pasty anhydrous product, aqueous phase, for example. It may be provided in the form of a dispersant, cream, skin, lotion, powder, ointment, spray or cone stick. It may also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.
본 발명의 화장품은 추가로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온봉쇄제 및 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 또는 화장품에 통상적으로 사용되는 임의의 다른 성분과 같은 화장품학 또는 피부과학 분야에서 통상적으로 사용되는 보조제를 함유할 수 있다. 그리고, 상기의 성분들은 피부과학 분야에서 일반적으로 사용되는 양으로 도입될 수 있다. Cosmetics of the present invention may further comprise fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionics. Such as type emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any other ingredients commonly used in cosmetics It may contain adjuvants conventionally used in the cosmetic or dermatology field. In addition, the above components may be introduced in an amount generally used in the dermatology field.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 최적화된 공정은 아니며, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, not an optimized process, it will be apparent to those of ordinary skill in the art that the scope of the present invention is not to be construed as being limited by these examples. will be.
실시예 1. 안정화된 라말린의 제조Example 1 Preparation of Stabilized Ramalin
1-1: 친수성 에어로겔의 제조1-1: Preparation of Hydrophilic Airgel
습윤겔 제조를 위하여, 먼저, 물, 금속 알콕사이드 용액으로 TMOS (Tetramethylortho- silicate)를, 촉매로는 시트릭산(citric acid, C6H8O7)과 NH4OH를 사용하였다. 첫번째 단계에서 원료의 혼합비율은 몰비로 TMOS : H2O : Methanol = 1 : 1.5 : 1 이며, 여기에 촉매로 시트릭산을 첨가하여 pH를 2.0으로 유지하며 상온/상압 상태에서 1시간 동안 교반시켜서 혼합하였다. 혼합 후 진공 증발기를 이용하여 메탄올을 제거하였다.To prepare the wet gel, first, TMOS (Tetramethylortho- silicate) was used as a water and metal alkoxide solution, and citric acid (C 6 H 8 O 7 ) and NH 4 OH were used as a catalyst. In the first step, the mixing ratio of the raw material is TMOS: H 2 O: Methanol = 1: 1.5: 1 in molar ratio, and by adding citric acid as a catalyst, the pH is maintained at 2.0 and stirred for 1 hour at room temperature / atmospheric pressure. Mixed. After mixing, the methanol was removed using a vacuum evaporator.
두 번째 단계에서는 TMOS 1몰 기준(첫번째 단계 TMOS의 양)으로 H2O 2.5몰과 희석용매로는 이소프로판올 (isopropanol) 500mL를, 촉매로는 NH4OH를 소량(이때 pH는 8.0～9.0을 유지하였다) 혼합하여 혼합용액을 제조하였다. pH가 높게 되면 실리카 분말 침전이 생성되어 겔이 형성되지 않는다. 이소프로판올 (isopropanol)의 양은 최종 친수성 에어로겔의 밀도를 조절하는 기능을 하므로 원하는 밀도에 맞추어 적정량을 넣는다. 즉 많이 넣으면 밀도가 낮아지고, 적게 넣으면 밀도가 높아진다. In the second step, 2.5 moles of H 2 O, 500 mL of isopropanol as the diluent solvent, and a small amount of NH 4 OH as the catalyst (the pH was maintained at 8.0∼9.0) based on 1 mole of TMOS (the amount of the first stage TMOS) Mixing to prepare a mixed solution. If the pH is high, silica powder precipitates and no gel is formed. The amount of isopropanol serves to control the density of the final hydrophilic airgel, so the appropriate amount is added to the desired density. If you put more, the density is lower, and if you put less, the density is higher.
두 번째 단계에서 제조된 용액 전체를 첫 번째 단계 용액에 함께 넣고 상온/상압에서 2시간 동안 혼합하였다. 두 번째 단계의 원료용액들을 첫 번째 단계에서 제조된 용액에 개별적으로 순차적으로 넣어선 안된다.The whole solution prepared in the second step was put together in the first step solution and mixed for 2 hours at room temperature / atmospheric pressure. The raw material solutions of the second stage should not be added individually and sequentially to the solution prepared in the first stage.
이상에서 제조된 졸용액을 밀폐용기에 부어 넣고 밀봉하여 50℃ 항온조에 넣고 겔화를 유도하였다. 일반적으로 상기의 조건이면 3시간 이내에 겔이 형성된다.The sol solution prepared above was poured into a sealed container, sealed, and placed in a 50 ° C. thermostatic bath to induce gelation. Generally, the gel is formed within 3 hours under the above conditions.
1-2: 초임계 건조공정 1-2: Supercritical Drying Process
1) 습윤겔 내의 용매 제거 및 치환1) Removal and Substitution of Solvent in Wet Gel
졸겔공정에서 제조된 습윤겔을 초임계 반응기에 넣고 반응기를 밀봉하였다. 고압액체 펌프를 이용하여 초임계 반응기 상부로 에탄올을 공급하여 상온에서 압력을 50기압으로 가압하였다. 50기압에 도달하면 초임계 반응기의 상부로 에탄올을 공급하며, 하부로 액체 혼합물을 방출시키며 50기압을 유지한다.(중력방향 이용) 이는 습윤겔 내에 잔류하는 촉매와 미반응 물(H2O)을 제거하기 위한 것이다. 이때 공급되는 에탄올의 총량은 초임계 반응기의 내부 용적의 두배 또는 습윤겔 부피의 세배 정도이며, 유속은 5mL/min이면 충분하다. The wet gel prepared in the sol-gel process was placed in a supercritical reactor and the reactor was sealed. Ethanol was supplied to the top of the supercritical reactor using a high pressure liquid pump to pressurize the pressure to 50 atm at room temperature. When 50 atm is reached, ethanol is fed to the top of the supercritical reactor, and the liquid mixture is discharged to the bottom and maintained at 50 atm (using gravity direction). This is the catalyst and unreacted water (H 2 O) remaining in the wet gel. It is to remove. In this case, the total amount of ethanol supplied is about twice the internal volume of the supercritical reactor or about three times the volume of the wet gel, and a flow rate of 5 mL / min is sufficient.
2) 습윤겔의 건조 2) drying of the wet gel
다음 단계로 액체 이산화탄소를 반응기의 상부에 공급하며 하부로 에탄올을 방출시켰다. 이때의 온도는 30℃이하, 압력은 70기압 이상을 반드시 유지하였다. In the next step, liquid carbon dioxide was fed to the top of the reactor and ethanol was released to the bottom. The temperature at this time was 30 degrees C or less, and the pressure necessarily maintained 70 or more atmospheres.
에탄올이 완전히 제거되면 반응기의 상부와 하부의 밸브를 잠그고 반응기의 히터를 가동시켜서 반응기 내부의 온도를 40℃ 까지 상승시키고, 이때 온도의 상승에 따라서 압력을 상승시킨다. 압력은 80∼100기압을 유지하였으며, 이보다 10기압 이상으로 압력이 상승하면 반응기 하부의 밸브를 조금 열어서 압력을 낮추어 조절한다.When the ethanol is completely removed, the valves of the upper and lower parts of the reactor are closed and the heater of the reactor is operated to raise the temperature inside the reactor to 40 ° C., at which time the pressure is increased as the temperature increases. The pressure was maintained at 80 to 100 atm, and when the pressure rises above 10 atm, the valve at the bottom of the reactor is opened to adjust the pressure.
위와 같이 초임계 상태에 도달한 후에는 반응기 내부의 온도를 40℃로 유지하며, 후방압력 조절기를 이용하여 이산화탄소를 방출시키고 이산화탄소 방출유속은 2기압/분 정도(0.2MPa/min)의 감압율을 유지하도록 하였으며 상압에 도달하면 냉각없이 반응기를 열고 건조된 친수성 에어로겔을 얻어냈다. After reaching the supercritical state as above, the temperature inside the reactor is maintained at 40 ° C, and carbon dioxide is released using a rear pressure regulator, and the carbon dioxide discharge flow rate is about 2 atm / min (0.2 MPa / min). When reaching atmospheric pressure, the reactor was opened without cooling to obtain a dried hydrophilic airgel.
회수된 친수성 에어로겔은 분쇄하여 원하는 크기의 분말로 제조하였다. The recovered hydrophilic airgel was ground to a powder of desired size.
1-3: 라말린의 합성1-3: Synthesis of Ramalin
상기 제조한 에어로젤에 흡착하기 위한 라말린은 다음과 같이 분리하였다. Ramalin for adsorption on the prepared airgel was separated as follows.
라말리나 테레브라타 (Ramalina terebrata)는 남극대륙 킹조지섬에 군락을 이루어 자생하는 지의류로서, 킹조지섬의 곳곳에서 용이하게 채취할 수 있다. Ramalina terebrata is a native lichen that colonizes King George Island in Antarctica and can be easily collected from all over King George Island.
완전히 동결 건조되고, 분쇄된 지의류 라말리나 테레브라타 샘플 672g을 메탄올:물 (5L, 80:20 v/v) 혼합용액을 이용하여 3회 반복하여 추출한 후, 동결건조를 거쳐 83g의 조추출물을 얻었다. 상기 조추출물을 1L의 증류수에 녹인 다음, 1L의 n-hexane과 클로로포름 (CHCl3)으로 추출하여 n-hexane 12.7g과 클로로포름 (CHCl3) 9.1g, 수용성 추출물 61.0g을 얻었다. 상기 수용성 추출물은 DPPH 자유 라디컬에 대해 높은 활성 IC50=9ug/ml를 나타냈다. 상기 수용성 추출물 중 5g을 이용하여 자동화된 MPLC (mild pressure liquid chromatography)를 적용시키고, 0%, 20%, 40%, 60%, 80%, 100% 메탄올-물 혼합용액으로 단계적으로 그래디언트 용매 시스템을 실시하였다. 0% 메탄올-물 혼합용액에 녹여진 추출물은 DPPH 자유 라디컬에 대해 높은 활성 IC50=8ug/ml를 나타냈고, 그 중 100mg 부분을 C18ODS 칼럼(250cm x 10cm)를 사용하여 준 분취 역상 HPLC(semi-preparative reverse phase HPLC)로 분석하였다. 사용한 용매 시스템은 0.1% 포름산이 섞인 물에 대해 0% 메탄올-물 혼합용액의 경우 10분, 20% 메탄올-물 혼합용액은 20분, 100% 메탄올용액에 대해서는 30분 이상 수행하였다. 유속은 2mL/분으로 하였다. After lyophilization of lyophilized and ground limalina ramera terretta sample was repeated three times using methanol: water (5L, 80:20 v / v) mixed solution, 83g of crude extract was obtained by lyophilization. Got it. The crude extract was dissolved in 1 L of distilled water and extracted with 1 L of n-hexane and chloroform (CHCl 3 ) to obtain 12.7 g of n-hexane, 9.1 g of chloroform (CHCl 3 ), and 61.0 g of an aqueous extract. The water soluble extract showed a high activity IC 50 = 9 ug / ml for DPPH free radicals. Automated Mild Pressure Liquid Chromatography (MPLC) was applied using 5 g of the aqueous extract, and the gradient solvent system was gradually phased with 0%, 20%, 40%, 60%, 80%, 100% methanol-water mixture solution. Was carried out. The extract dissolved in a 0% methanol-water mixture showed a high activity IC 50 = 8 ug / ml for DPPH free radicals, of which 100 mg portion was subjected to semipreparative reversed phase using a C 18 ODS column (250 cm x 10 cm). It was analyzed by HPLC (semi-preparative reverse phase HPLC). The solvent system used was 10 minutes for a 0% methanol-water mixed solution for water mixed with 0.1% formic acid, 20 minutes for a 20% methanol-water mixed solution and 30 minutes for a 100% methanol solution. The flow rate was 2 mL / min.
그 결과, 화합물은 자외선 280nm에서 흡수되었다. 다섯 번째 분획 (45mg; tR = 18.88 분)의 경우, DPPH 자유 라디칼에 대해 가장 높은 활성 IC50=1ug/ml를 나타냈으며, 그 분획을 C18ODS 칼럼 (250cm x 10cm)를 사용하여 준 분취 역상 HPLC (semi-preparative reverse phase HPLC)로 반복하여 정제하였다. 그래디언트 용매 시스템은 10~30% 아세토니트릴 (acetonitrile) 수용액 (0.1% 포름산)을 사용하여 50분 이상 수행하였으며, 유속은 2mL/분이었다. 그 결과, 8.26분에 DPPH 자유라디칼에 대한 활성이 IC50=0.99ug/ml인 화학식 1로 표시되는 라말린 30mg을 얻었다. As a result, the compound was absorbed at 280 nm ultraviolet. For the fifth fraction (45 mg; tR = 18.88 min), the highest active IC 50 = 1 ug / ml was shown for DPPH free radicals, which fraction was subfractional reversed using a C 18 ODS column (250 cm x 10 cm). Purification was repeated by HPLC (semi-preparative reverse phase HPLC). The gradient solvent system was run for at least 50 minutes using 10-30% acetonitrile aqueous solution (0.1% formic acid) and the flow rate was 2 mL / min. As a result, at 8.26 minutes, 30 mg of RAMALIN represented by the formula (1) having an activity of DPPH free radicals of IC 50 = 0.99 ug / ml was obtained.
1-4: 라말린 흡착공정1-4: Ramalline adsorption process
40℃로 가온된 정제수 100g에 친수성 에어로겔 23.3g을 천천히 투입하면서 24시간 동안 교반 하였다(A상). 40℃로 가온된 정제수에 실시예 1-3에서 얻은 라말린 10g을 넣고 용해한 혼합물을 A상에 천천히 적가 후 25℃를 유지하면서 12시간 동안 교반 하였다. 이 혼합물에 아세톤 2㎏을 천천히 적가 하여 친수성 에어로겔에 라말린의 흡착을 촉진한 후 진공 여과를 통하여 슬러리를 수득 후 45℃에서 12시간 동안 진공건조 하여 라말린-친수성 에어로겔을 31.8g 얻었다.23.3 g of hydrophilic airgel was slowly added to 100 g of purified water warmed to 40 ° C. and stirred for 24 hours (phase A). 10 g of ramalin obtained in Example 1-3 was added to purified water warmed to 40 ° C., and the dissolved mixture was slowly added dropwise to A phase, followed by stirring for 12 hours while maintaining 25 ° C. 2 kg of acetone was slowly added dropwise to the mixture to promote the adsorption of ramalin to the hydrophilic airgel, and a slurry was obtained by vacuum filtration, followed by vacuum drying at 45 ° C. for 12 hours to obtain 31.8 g of ramine-hydrophilic airgel.
1-5: 라말린-친수성 에어로겔 표면처리1-5: Ramallin-Hydrophilic Airgel Surface Treatment
50℃로 가온된 에탄올 50g에 레시친 3.1g을 용해한 후 실시예 1-4에서 얻은 라말린-친수성 에어로겔 31g을 천천히 넣으면서 아지믹서를 이용하여 골고루 분산 후 60℃에서 감압증류 하여 표면처리된 라말린-친수성 에어로겔 파우더 32g을 얻었다.After dissolving 3.1 g of lecithin in 50 g of ethanol heated to 50 ° C., 31 g of the ramalin-hydrophilic aerogel obtained in Example 1-4 was slowly added, dispersed evenly using an azimixer, and then distilled under reduced pressure at 60 ° C. to give a surface treatment. 32 g of hydrophilic airgel powder was obtained.
1-6: 서브마이크로 입자의 현탁액 제조1-6: Preparation of Suspension of Submicro Particles
카프릴릭/카프릭트리글리세라이드 210g에 실시예 1-5에서 얻은 파우더를 28g 넣고 60℃로 가온하면서 글리세린 357g을 투입 후 70℃로 가온하였고, 여기에 레시친 35g을 넣고 완전 용해하였다. 이 혼합물에 정제수 70g을 투입 후 4,000rpm으로 20분간 교반 후 고압균질기 (microfludizer)를 이용하여 1,000bar에서 3회 통과 시켰다.28 g of the powder obtained in Example 1-5 was added to 210 g of caprylic / capric triglyceride, and 357 g of glycerin was added while warming to 60 ° C., and warmed to 70 ° C., and 35 g of lecithin was added thereto to completely dissolve it. After adding 70 g of purified water to the mixture and stirring at 4,000 rpm for 20 minutes, the mixture was passed three times at 1,000 bar using a high pressure homogenizer (microfludizer).
실시예 2: 안정화된 라말린의 안정성 시험Example 2: Stability Test of Stabilized Ramalin
실시예 1에서 제조된 안정화 라말린의 안정성을 평가하기 위하여 하기와 같이 시험하였다.The following test was conducted to evaluate the stability of the stabilized ramine prepared in Example 1.
＊검체보관조건: 25, 45℃ (안정화 라말린)＊ Storage condition: 25, 45 ℃ (stabilized ramaline)
＊분석 조건＊ Condition condition
사용기기: HPLC (Prostar, Varian, Holland)Instrument used: HPLC (Prostar, Varian, Holland)
검출기: UV/Vis detector 235 nmDetector: UV / Vis detector 235 nm
컬럼: Mightsil RP-18 GP 250-4.6 (5 um, Kanto, Japan)Column: Mightsil RP-18 GP 250-4.6 (5 um, Kanto, Japan)
이동상: 0.01M KH2PO4 (pH2.5) : MeOH ( 90 : 10 )Mobile phase: 0.01M KH 2 PO 4 (pH2.5): MeOH (90: 10)
유속: 1 mL/min.Flow rate: 1 mL / min.
라밀린 표준품 및 안정화된 라말린을 각각 10 mg/L의 농도가 되도록 이동상에 용해 후 각각을 상기 분석조건에 따라 분석 후 면적값을 아래의 식처럼 비교하여 농도를 산출하였으며, 안정성은 초기값을 100%로 하여 결과를 비교하였다.After dissolving the ramyl standard and the stabilized ramalline to the concentration of 10 mg / L, respectively, and after analyzing each of them according to the above analysis conditions, the concentration was calculated by comparing the area values as shown below. The results were compared at 100%.
안정성 계산식Stability Formula
Figure PCTKR2013002247-appb-I000003
Figure PCTKR2013002247-appb-I000003
그 결과, 도 4에 나타난 바와 같이, 안정화된 라말린 용액과 동일 농도로 제조된 라말린 수용액을 25℃와 45℃의 인큐베이터에 보관하면서 1주일 간격으로 4주 동안 관찰한 결과, 안정화 라말린 용액은 25℃에서 보관 시 4주 후에도 약 96%의 높은 역가 유지율을 보이는 반면 비안정화 라말린 수용액은 1주만에 역가의 50% 이상이 손실 되었다. 또한, 안정화 라말린 용액은 45℃에서는 61%의 역가 유지율을 보여 주었고, 비안정화 라말린 수용액은 45℃에서 1주일만에 전체 역가의 90% 이상이 파괴되었다. 따라서 상기 라말린 안정화 방법은 라말린의 안정성 향상에 크게 도움을 주는 것으로 나타났다.As a result, as shown in Figure 4, the stabilizing ramalin solution was prepared at the same concentration as the stabilizing ramalin solution in the incubator at 25 ℃ and 45 ℃ observed for four weeks at intervals of one week, stabilizing ramalin solution Was maintained at a high titer of about 96% even after 4 weeks when stored at 25 ℃, while unstable aqueous solution of unstable ramaline lost more than 50% of the titer in one week. In addition, the stabilized ramalin solution showed a titer retention rate of 61% at 45 ° C., and the unstabilized ramalin aqueous solution destroyed more than 90% of the total titer at 45 ° C. in one week. Therefore, the method of stabilizing the RAMALIN has been shown to greatly aid in improving the stability of RAMALIN.
실시예 3: 안정화된 라말린의 화장료 원료로써의 효능 시험Example 3 Efficacy Test of Stabilized Ramalin as a Cosmetic Ingredient
실시예 1에서 제조된 안정화 라말린의 화장료 원료로써의 효능을 평가하기 위하여 하기와 같이 시험하였다.In order to evaluate the efficacy of the stabilized RAMALIN prepared in Example 1 as a cosmetic raw material was tested as follows.
실시예 1에서 제조된 안정화 라말린을 포함하는 화장료 제형을 하기의 표 1과 같이 제조한 후, 20∼60세의 여성 피검자 30명을 두 그룹으로 나누어 라말린이 포함된 화장료 및 대조구 화장료를 매일 아침 저녁 약 1 g을 안면부에 4주간 도포하게 한 후 매 주 피검자의 안면부 피부 밝기 및 주름 개선 정도를 평가하였다.After preparing a cosmetic formulation comprising the stabilized ramallin prepared in Example 1 as shown in Table 1 below, divided 30 groups of 20 to 60-year-old female subjects into two groups daily cosmetic and control cosmetics containing ramalin Approximately 1 g was applied to the face for 4 weeks in the morning and evening, and then the subject's facial skin brightness and wrinkle improvement were evaluated every week.
표 1
Figure PCTKR2013002247-appb-T000001
 Table 1
Figure PCTKR2013002247-appb-T000001
피부 밝기의 개선 정도는 ARAMO TS (아람 휴비스, 대한민국)를 사용하여 측정하였으며, 측정된 값의 변화는 하기의 표 2에 나타내었다. 아울러, 주름 개선효과는 실험이 시작되기 전 피검자 눈 밑의 레플리카 (replica)를 채취하고 (Xantopren, Bayer), 실험이 종료된 4주 후 레플리카를 눈밑의 동일 부위에서 채취하여 영상분석을 통해 주름의 2차원적 분석으로 주름의 밀도를 측정하였으며, 영상분석에 의한 주름 밀도의 측정 결과는 사용 전에 대한 주름 밀도에 대한 감소율을 평균하여 표 3에 나타내었다.The degree of improvement of skin brightness was measured using ARAMO TS (Aram Huvis, South Korea), and the change in the measured value is shown in Table 2 below. In addition, the wrinkle improvement effect was obtained by taking a replica under the subject's eyes (Xantopren, Bayer) before the experiment began, and after 4 weeks after the experiment was completed, a replica was taken from the same area under the eyes to analyze the wrinkles. Wrinkle density was measured by two-dimensional analysis, and the results of the measurement of wrinkle density by image analysis are shown in Table 3 by averaging the reduction ratio of the wrinkle density before use.
표 2
Figure PCTKR2013002247-appb-T000002
 TABLE 2
Figure PCTKR2013002247-appb-T000002
표 3
Figure PCTKR2013002247-appb-T000003
 TABLE 3
Figure PCTKR2013002247-appb-T000003
상기 표 2 및 3에 나타난 바와 같이, 본 발명에 의한 안정화된 라말린 용액을 포함하는 화장료 조성물을 도포한 경우 미백효과가 있으며, 주름의 밀도가 사용전보다 크게 감소하는 것을 확인할 수 있었다. 아울러, 피검자 모두 피부결의 개선되었음을 보고한바 본 물질이 주름 및 피부결 개선에 도움을 주는 물질임을 확인할 수 있었다.As shown in Tables 2 and 3, when the cosmetic composition containing the stabilized ramine solution according to the present invention was applied, there was a whitening effect, and it was confirmed that the density of the wrinkles was greatly reduced than before use. In addition, all the subjects reported that the skin texture was improved, and it was confirmed that the material was helpful for improving wrinkles and skin texture.
본 발명에 따른 라말린의 안정화 방법은 장기간 보관시에도 라말린 표준품에 대비하여 거의 유사한 역가를 유지하도록 라말린을 안정화시킬 수 있어 항산화 및 항염증 효과가 뛰어난 라말린을 장기간 보관할 수 있게 하는 장점이 있다. 아울러, 본 발명에 따른 방법은 45℃와 같은 고온에서도 장기간 보관 시 역가가 급격히 낮아지지 않고 높은 수준으로 유지시킬 수 있어 라말린의 보관을 용이하게 하는 점에서 유용하다. 아울러, 본 발명에 따른 안정화된 라말린 용액은 피부의 밝기를 개선시키고 주름을 개선하는 효과가 있는바 미백 및 주름 개선용 기능성 화장품으로서 사용될 수 있다.The stabilization method of ramalin according to the present invention is capable of stabilizing ramalin so as to maintain a nearly similar titer compared to the ramalin standard even in long-term storage, which has the advantage of allowing the long-term storage of ramin excellent in antioxidant and anti-inflammatory effects. have. In addition, the method according to the present invention is useful in that it can be maintained at a high level without rapidly lowering the titer even when stored at a high temperature such as 45 ° C. to facilitate the storage of ramalin. In addition, the stabilized RAMALIN solution according to the present invention can be used as functional cosmetics for whitening and wrinkle improvement as it has an effect of improving the brightness of the skin and improving wrinkles.
이상으로 본 발명의 내용의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.As described above in detail a specific part of the content of the present invention, for those skilled in the art, such a specific description is only a preferred embodiment, which is not limited by the scope of the present invention Will be obvious. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (13)

  1. 다음 단계를 포함하는 하기 화학식 1로 표시되는 라말린의 안정화 방법:Method for stabilizing Ramalin represented by the following formula (1) comprising the following steps:
    (a) 친수성의 다공성 매트릭스에 라말린을 흡착시키는 단계;(a) adsorbing ramalin to a hydrophilic porous matrix;
    (b) 상기 라말린이 흡착된 다공성 매트릭스의 표면을 소수성으로 개질하는 단계; 및(b) hydrophobically modifying the surface of said ramal adsorbed porous matrix; And
    (c) 상기 소수성으로 개질된 다공성 매트릭스를 유기 용매에 분산시키는 단계(c) dispersing the hydrophobically modified porous matrix in an organic solvent
    화학식 1Formula 1
    Figure PCTKR2013002247-appb-I000004
    Figure PCTKR2013002247-appb-I000004
  2. 제1항에 있어서, 상기 친수성의 다공성 매트릭스는 친수성 에어로겔로 구성되는 것을 특징으로 하는 방법. The method of claim 1 wherein the hydrophilic porous matrix consists of a hydrophilic airgel.
  3. 제2항에 있어서, 상기 친수성 에어로겔은 물, 알콜 및 금속 알콕사이드를 원료로 하여 제조된 것임을 특징으로 하는 방법.The method according to claim 2, wherein the hydrophilic airgel is prepared using water, alcohol and metal alkoxide as raw materials.
  4. 제2항에 있어서, 상기 친수성 에어로겔은 초임계 유체 건조 공정을 이용하여 제조된 것임을 특징으로 하는 방법. 3. The method of claim 2, wherein the hydrophilic airgel is prepared using a supercritical fluid drying process.
  5. 제2항에 있어서, 상기 친수성 에어로겔은 시트르산 촉매 하에서 테트라메틸-오소-실리케이트 (tetramethylortho-silicate): 물: 메탄올을 1: 1.5:1의 비로 혼합하여 교반한 혼합물에, NH4OH 촉매 하에서 테트라메틸-오소-실리케이트:물을 이소프로판올을 희석용매로 하여 1: 2.5의 비로 혼합한 용액을 가하여 겔을 형성한 다음, 용매를 제거하고 건조하여 제조된 것임을 특징으로 하는 방법. The method of claim 2, wherein the hydrophilic aerogel is tetramethyl- ortho-silicate (tetramethylortho-silicate): water: methanol in a mixture of 1: 1.5: 1 in a mixture stirred, tetramethyl- ortho under NH 4 OH catalyst -Silicate: water is prepared by adding a solution of isopropanol as a dilution solvent in a ratio of 1: 2.5 to form a gel, then removing the solvent and drying.
  6. 제1항에 있어서, 상기 (a) 단계는 친수성의 다공성 매트릭스를 수용성 용매에 침지시킨 후 화학식 1로 표시되는 라말린이 용해되어 있는 수용성 용액을 첨가함으로써 흡착시키는 것을 특징으로 하는 방법. The method of claim 1, wherein the step (a) is characterized in that the hydrophilic porous matrix is immersed in an aqueous solvent and then adsorbed by adding an aqueous solution in which the ramalin represented by Formula 1 is dissolved.
  7. 제1항에 있어서, 상기 (b) 단계는 양쪽성 계면활성제 또는 계면활성제 화합물을 처리하여 소수성으로 개질하는 것을 특징으로 하는 방법. The method of claim 1, wherein step (b) is characterized in that to modify the hydrophobic by treating the amphoteric surfactant or surfactant compound.
  8. 제7항에 있어서, 상기 양친매성 화합물은 레시틴, 포스파티딜 에탄올아민, 리소레시틴, 아미노프로필 라우릴글루타민(aminopropyl laurylglutamine), 다이소듐 탈로우암포다이아세테이트 (Disodium tallowam- phodiacetate), 코코암포다이프로피오닉 애씨드(Cocoamphodipropionic acid), 다이소듐 카프로암포다이아세테이트(Disodium Caproamphodiacetate), 코카미도프로필 베타인(Cocamidopropyl Betain), 소듐 C12-15 알콕시프로필 이미노프로피오네이트(Sodium C12-15 alkoxypropyl Iminodipropionate) 및 쿼터니움-85(Quaternium-85)로 구성된 군에서 선택되는 것을 특징으로 하는 방법. The method of claim 7, wherein the amphiphilic compound is lecithin, phosphatidyl ethanolamine, lysocithin, aminopropyl laurylglutamine, disodium tallowam- phodiacetate, coco ampodipropionic acid (Cocoamphodipropionic acid), Disodium Caproamphodiacetate, Cocamidopropyl Betain, Sodium C12-15 alkoxypropyl Iminodipropionate and Quaternium-85 (Quaternium-85).
  9. 제1항에 있어서, 상기 (c) 단계의 유기용매는 카프릴릭/카프릭트리글리세라이드, 스쿠알란, 실리콘 오일, 미네랄 오일, 식물성 오일 및 이들의 혼합물로 구성된 군에서 선택되는 것을 특징으로 하는 방법. The method of claim 1, wherein the organic solvent of step (c) is selected from the group consisting of caprylic / capric triglyceride, squalane, silicone oil, mineral oil, vegetable oil, and mixtures thereof.
  10. 제1항 내지 제9항 중 어느 한 항의 방법에 의하여 제조되는 것을 특징으로 하는 안정화된 라말린 용액.10. A stabilized ramalline solution, prepared by the method of any one of claims 1-9.
  11. 제10항의 안정화된 라말린 용액을 유효성분으로 함유하는 미백 또는 주름 개선용 화장료 조성물.A cosmetic composition for improving whitening or wrinkles, comprising the stabilized ramalene solution of claim 10 as an active ingredient.
  12. 제11항에 있어서, 상기 안정화된 라말린 용액은 화장료 조성물 총 중량에 대하여 0.0001% 내지 10%로 포함되는 것을 특징으로 하는 화장료 조성물.The cosmetic composition according to claim 11, wherein the stabilized ramalin solution is contained in an amount of 0.0001% to 10% based on the total weight of the cosmetic composition.
  13. 제11항에 있어서, 상기 화장료 조성물은 화장수, 유액, 크림, 에센스, 화장연고, 스프레이, 젤, 팩, 선 스크린, 메이크업 베이스, 파운데이션, 파우더, 메이크업 세정제 및 세정제로 구성되는 군에서 선택된 제형을 갖는 것을 특징으로 하는 화장료 조성물.The cosmetic composition of claim 11, wherein the cosmetic composition has a formulation selected from the group consisting of lotion, latex, cream, essence, cosmetic ointment, spray, gel, pack, sunscreen, makeup base, foundation, powder, makeup cleaner, and cleaner. Cosmetic composition, characterized in that.
PCT/KR2013/002247 2012-03-19 2013-03-19 Method for stabilizing ramalin using porous matrix, and stablized ramalin solution WO2013141576A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020120027649A KR101429242B1 (en) 2012-03-19 2012-03-19 Stabilization Method of Ramalin Using Porous Matrix and Stabilized Ramalin Solution
KR10-2012-0027649 2012-03-19

Publications (1)

Publication Number Publication Date
WO2013141576A1 true WO2013141576A1 (en) 2013-09-26

Family

ID=49222963

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2013/002247 WO2013141576A1 (en) 2012-03-19 2013-03-19 Method for stabilizing ramalin using porous matrix, and stablized ramalin solution

Country Status (2)

Country Link
KR (1) KR101429242B1 (en)
WO (1) WO2013141576A1 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102631297B1 (en) * 2021-11-09 2024-01-30 주식회사에이치엔비랩스 Manufacturing method of nanosome with improved stability through surface treatment

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010053327A2 (en) * 2008-11-10 2010-05-14 한국해양연구원 Novel compound ramalin, and use thereof
WO2011152671A2 (en) * 2010-06-03 2011-12-08 한국해양연구원 Pharmaceutical composition for the prevention or treatment of inflammatory diseases or immune diseases containing ramalin
WO2012008785A2 (en) * 2010-07-14 2012-01-19 한국해양연구원 Method for preparing ramalin

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2195058B1 (en) * 2007-08-30 2019-02-27 Baska, Meenakshi Laryngeal mask

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010053327A2 (en) * 2008-11-10 2010-05-14 한국해양연구원 Novel compound ramalin, and use thereof
WO2011152671A2 (en) * 2010-06-03 2011-12-08 한국해양연구원 Pharmaceutical composition for the prevention or treatment of inflammatory diseases or immune diseases containing ramalin
WO2012008785A2 (en) * 2010-07-14 2012-01-19 한국해양연구원 Method for preparing ramalin

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
BHATTARAI, H. D. ET AL.: "thin layer chromatography analysis of antioxidant constituents of lichens from Antarctica", JOURNAL OF NATURAL MEDICINES, vol. 62, 2008, pages 481 - 484, XP055157233, DOI: doi:10.1007/s11418-008-0257-9 *
GULLUCE, M. ET AL.: "Screening the antioxidant and antimicrobial properties of the lichens Parmelia saxatilis, Platismatia glauca, Ramalina pollinaria, Ramalina polymorpha and Umbilicaria nylanderiana", PHYTOMEDICINE, vol. 13, 2006, pages 515 - 521, XP028022214, DOI: doi:10.1016/j.phymed.2005.09.008 *
KINOSHITA, T. ET AL.: "The synthesis of Anthglutin and its analogues", BULLETIN OF THE CHEMICAL SOCIETY OF JAPAN, vol. 44, 1981, pages 2219 - 2220 *

Also Published As

Publication number Publication date
KR101429242B1 (en) 2014-08-12
KR20130106045A (en) 2013-09-27

Similar Documents

Publication Publication Date Title
WO2019235840A2 (en) Fermented emulsifier for forming reverse micelle, composition comprising same and method for producing fermented emulsifier
US9067967B2 (en) Peptides useful in the treatment and care of the skin and mucous membranes and their use in cosmetic or pharmaceutical compositions
EP2646115B1 (en) Exopolysaccharide for the treatment and/or care of the skin, mucous membranes, hair and/or nails
KR102217951B1 (en) Exopolysaccharide for the treatment and/or care of the skin, mucous membranes and/or nails
WO2020096156A1 (en) Novel lactobacillus gasseri skb1102 strain or cosmetic composition containing same having anti-pollution function
US20200002377A1 (en) Compounds useful for the treatment and/or care of the skin, hair, nails and/or mucous membranes
JPH0694449B2 (en) Cosmetic composition
WO2011162478A2 (en) Cosmetic composition containing retinol stabilized by porous polymer beads and nanoemulsion
EP2579952A2 (en) Skin antiaging treatment
KR101188643B1 (en) Vesicles of high molecular weight silicone polyethers
WO2012173458A2 (en) Stabilized active materials contained in plant cells and method for preparing same
WO2024136468A1 (en) Pigment prepared by using microorganisms and method for preparing same
WO2013141576A1 (en) Method for stabilizing ramalin using porous matrix, and stablized ramalin solution
US20130078295A1 (en) Skin antiaging treatment
JP2009528348A (en) Silicone vesicles containing active agents
KR102263449B1 (en) High purity fermented-surfactant and preparation method thereof
WO2019132070A1 (en) Antiaging cosmetic containing nanoparticle soluble product obtained by solubilizing poorly soluble oleanolic acid, and method for preparing antiaging cosmetics
WO2020085857A1 (en) Transdermal delivery system comprising reverse micelle
WO2019045515A1 (en) Oil-in-water type cosmetic composition having stabilized high-content oil inner phase
KR100858449B1 (en) Cosmetic compositions comprising extract of viola tricolor l. stabilized in nano-liposome
WO2023085578A1 (en) Method for preparing fermented vegetable oil by means of skin flora, and cosmetic composition comprising same
WO2019139214A1 (en) Cosmetic composition containing algasome comprising microalgae extract therein
KR101096374B1 (en) Method of stabilization of active ingredients using polyol/polymer microcapsules and cosmetic composition containing the microcapsules
KR101768029B1 (en) Cosmetic Composition For Moisturizing Skin Comprising Perna Canaliculus as Active Ingredient
KR101356796B1 (en) Liposome Collecting Kaempferitrin and Coating Carrageenan and Method of The Same

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 13763679

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A) DATED 30/01/2015)

122 Ep: pct application non-entry in european phase

Ref document number: 13763679

Country of ref document: EP

Kind code of ref document: A1