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WO2015099274A1 - Structure dans laquelle une matière active est introduite dans un protoplaste végétal dé-différencié, son procédé de préparation et composition cosmétique la contenant - Google Patents

Structure dans laquelle une matière active est introduite dans un protoplaste végétal dé-différencié, son procédé de préparation et composition cosmétique la contenant Download PDF

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Publication number
WO2015099274A1
WO2015099274A1 PCT/KR2014/010012 KR2014010012W WO2015099274A1 WO 2015099274 A1 WO2015099274 A1 WO 2015099274A1 KR 2014010012 W KR2014010012 W KR 2014010012W WO 2015099274 A1 WO2015099274 A1 WO 2015099274A1
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Prior art keywords
plant
active material
cosmetic composition
cells
active substance
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English (en)
Korean (ko)
Inventor
심성보
박소현
홍우진
최은호
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Coway Co Ltd
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Coway Co Ltd
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Priority to CN201480071246.6A priority Critical patent/CN105848635B/zh
Publication of WO2015099274A1 publication Critical patent/WO2015099274A1/fr
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    • 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
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • 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
    • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A61K8/4906—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom
    • A61K8/4926—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom having six membered rings
    • 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
    • A61K8/67—Vitamins
    • 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
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9771—Ginkgophyta, e.g. Ginkgoaceae [Ginkgo family]
    • 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
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • 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
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9794—Liliopsida [monocotyledons]
    • 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
    • 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
    • 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
    • A61K2800/56—Compounds, absorbed onto or entrapped into a solid carrier, e.g. encapsulated perfumes, inclusion compounds, sustained release forms
    • 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
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95

Definitions

  • the present invention relates to a structure in which an active substance is introduced into a dedifferentiated plant protoplast, a preparation method thereof, and a cosmetic composition comprising the same as an active ingredient in order to increase the stability and efficacy of the active substance.
  • phyto-chemicals inherent in plants that exist in nature, and there are a myriad of them, including terpenoids, flavonoids, flavonols, polyphenols, amino acids, lignans, alkaloids, vitamins and catechin compounds. consist of.
  • Patent Laid-Open No. 2012-102136 discloses a cosmetic composition comprising a lyophilized product of dedifferentiated plant cells of a basophilic plant such as Criste Marine for skin bleaching and lightening.
  • an effective active substance such as retinol and the like has a useful function in the physiological characteristics in the human body, but is not free from stability, various methods have been proposed to increase its stability.
  • liposomes using phospholipids or surfactants such as liposomes using phospholipids or surfactants, stabilized cerasomes using ceramides, liquid crystals stabilized by liquid crystal structures, and cubic cubosomes using monoglycerides.
  • International Patent WO2012-173458 discloses a cosmetic composition containing plant cells stabilized with active substances such as gallic acid, amino acids, etc., wherein the plant cells are cell walls, and the active substances are introduced into the cell walls. It was suggested that the stability of the active substance can be increased.
  • the cell wall of the plant cell contains excess cellulose, which is not easy to penetrate other substances, so it needs to be removed for the entry of the active substance, and if the cell wall and the cell membrane are removed, the stability of the introduced active substance is rather deteriorated. Can be.
  • Patent Document 1 International Patent WO2013-180526
  • Patent Document 2 International Patent WO2012-102456
  • Patent Document 3 International Patent WO2009-139581
  • Patent Document 4 International Patent WO2012-173458
  • the present inventors have conducted various studies to stably capture active substances in plant cells at high inlet rate. As a result, the present inventors have removed the cell wall of plant cells and used protoplasts composed of cell membranes and small cells. The present invention was completed by confirming that it can be used as a cosmetic composition by increasing the penetration rate of the active substance as well as the permeability to.
  • an object of the present invention is to provide a structure and a method of manufacturing the active material is introduced at a high pulling rate, which can maximize the efficacy of the active material.
  • Another object of the present invention to provide a cosmetic composition comprising the structure as an active ingredient.
  • the present invention provides a structure in which the active material is introduced into the dedifferentiated plant protoplast.
  • a method of making a structure in which an active substance is introduced into a dedifferentiated plant protoplast is provided.
  • the present invention also provides a cosmetic composition
  • a cosmetic composition comprising as an active ingredient a structure in which an active substance is introduced into the dedifferentiated plant protoplast.
  • the structure according to the present invention maintains the stability of the active ingredient existing in the plant cell intact and additionally has high stability of the active substance introduced into the cell, and the efficacy by the active substance is also improved, preferably as an active ingredient composition of the cosmetic Can be used
  • FIG. 1 is a flow chart showing the steps of producing a structure in which an active substance is introduced into a dedifferentiated plant protoplast according to the present invention.
  • Figure 2 (a) is a standard sample of cyclohexanediol bisethylhexanoate, (b) is a GC spectrum of the structure in which cyclohexanediol bisethylhexanoate is introduced into the prototype of Example 1
  • Figure 3 (a) is a standard sample bisretin amido methyl pentane, (b) is a GC spectrum of the structure in which the bisretin amido methyl pentane is introduced into the propoflast of Example 2
  • Figure 4 (a) is beta rapachon is a standard sample, (b) is a GC spectrum of the structure in which beta rapachon is introduced into the propoflast of Example 3
  • Figure 5 (a) is a growth factor complex peptide of the standard sample, (b) is a MALDI-TOF mass spectrogram of the structure in which the growth factor complex peptide is inserted in the propoflast of Example 4
  • Example 6 is a graph showing the effect of inhibiting the production of melanin at each concentration of the structure of Example 1, cyclohexanediol bisethylhexanoate and alpha-bisabolol
  • Example 7 is a graph showing collagen synthesis ability of the structure of Example 2, bisretinamido methylpentane, retinol and adenosine
  • FIG. 8 is a graph showing collagen synthesis ability of the construct, betarapacachon, retinol and adenosine of Example 3.
  • FIG. 8 is a graph showing collagen synthesis ability of the construct, betarapacachon, retinol and adenosine of Example 3.
  • Figure 9 is a graph showing the collagen synthesis of the structure of Example 4, growth factor complex peptide, retinol and adenosine
  • Example 10 is an image showing epidermal regeneration before and after treatment of the construct of Example 4 on human skin tissue
  • Example 11 is an image showing the degree of glycosaminoglycan increase before and after treatment of the construct of Example 4 for human skin tissue
  • the present invention provides a structure in which an active substance is introduced into a dedifferentiated plant protoplast.
  • Protoplasm is a protoplast in which a cell wall of a cell is removed and a cell membrane is present, and various active substances which can be used as a cosmetic composition in the cell membrane can be introduced therein.
  • the cell wall contains an excess of cellulose, which is not easy to penetrate other materials, so it needs to be removed for the introduction of the active material, and if the cell wall and the cell membrane are removed, the stability of the introduced active material may be lowered. Therefore, protoplasts with cell membranes are preferred.
  • the active substance may be a hydrophilic substance or a hydrophobic substance, and is not particularly limited in the present invention.
  • Representative examples of the active substance include organic acids, vitamins, arbutin, adenosine, niacinamide, polyphenols, flavonoids, retinol, cyclohexanediol bisethylhexanoate, bisretinamido methylpentane, betarapach, growth factor, growth One kind selected from the group consisting of factor complex peptides, and combinations thereof is possible.
  • the active substance is itself used as an active ingredient of the cosmetic composition, it is not stable enough to express its activity, but it is incorporated into the protoplast, thereby increasing the stability of the activity, and the biocompatibility is also greatly improved due to the protoplast. There is an advantage.
  • Figure 1 is a flow chart showing the steps of manufacturing a structure in which the active material is introduced into the dedifferentiated plant protoplast according to the present invention, which will be described in detail with respect to each step with reference to Figure 1 below:
  • step (a) the cells of the plant are obtained from the leaves, stems, roots, flowers, berries and seeds of the plant.
  • Plant cell harvesting is not particularly limited in the present invention, and known methods can be used.
  • the embodiment of the present invention after sterilization, soaked in an aqueous solution of sodium chlorate added with a surfactant and then washed to obtain plant cells.
  • curry plant Helichrysum italicum
  • gugal Commiphora wightii, Commiphora myrrha
  • prickly pear Opuntia Ficus indica
  • peony Paeonia lactiflora
  • petrified Adenium obesum
  • Nymphia ordorata Nymphaea coerulea
  • Eucalyptus punctata Ginkgo ( Ginkgo biloba ), Lilium candidum , Olea europaea
  • Papyrus Cyperus papyrus
  • Lotus Nelumbo nucifera
  • Redwood Sequoia sempervirens
  • Rosa gallica officinalis Coffea arabica
  • Plumeria obtusa Plumeria obtusa
  • Gardenia jasminoides Bougainvillea spectabilis
  • step (b) the obtained plant cells are dedifferentiated using auxin to obtain dedifferentiated plant cells.
  • auxin which is used for dedifferentiation, is one of the plant growth regulators, which promotes cell elongation at low concentrations but inhibits growth at high concentrations.
  • the auxin one species selected from the group consisting of alpha-naphtalene acetic acid, 2,4-dichlorophenoxy acetic acid, indole-3-acetic acid, and combinations thereof It is possible to induce dedifferentiation of the plant cells by culturing the plant cells in a medium containing them at a concentration of 1 to 5 mg / l.
  • the medium is not particularly limited in the present invention, any medium used for plant cell culture may be used.
  • step (c) the dedifferentiated plant cells obtained in step (b) are cultured in large quantities.
  • Mass cultivation of dedifferentiated plant cells is carried out in a variety of media, wherein as the medium may be a known medium such as MS medium, B5 medium, WHITE medium, N6 medium, SH medium, Anderson medium, preferably MS Use the medium.
  • the medium may be a known medium such as MS medium, B5 medium, WHITE medium, N6 medium, SH medium, Anderson medium, preferably MS Use the medium.
  • culture conditions and period are not particularly mentioned in the present invention, it can be carried out under the conditions as known.
  • step (d) the cell walls of the largely cultured dedifferentiated plant cells are removed using an enzymatic reaction to obtain a protoplast remaining only in the cell membrane and organelles.
  • the enzyme consists of cellulase (EC 3.2.1.4), pectinase (EC 3.2.1.15), xylanase (EC 3.2.1.8), chitinase (EC 3.2.1.14), hemicellulase and combinations thereof
  • cellulase EC 3.2.1.4
  • pectinase EC 3.2.1.15
  • xylanase EC 3.2.1.8
  • chitinase EC 3.2.1.14
  • hemicellulase hemicellulase
  • These enzymes can be used in various concentrations of 0.01 to 10% by weight, preferably 0.1 to 5% by weight of cellulase, 0.01 to 2.5% by weight of pectinase, 0.1 to 5% by weight of hemicellulase, more preferably A multicomponent enzyme mixture of 1% by weight cellulase, 2% by weight hemicellulase and 0.5% by weight pectinase is used.
  • the enzymatic reaction is carried out in a temperature range of 4 ° C. to 40 ° C. (more preferably 10 ° C. to 25 ° C.) at a rate of 10 to 50 rpm for 15 hours to 24 hours. Only the cell wall is removed and the cell membrane can be stably preserved.
  • step (e) the active material is introduced into the obtained dedifferentiated plant protoplasm by a pressure osmotic process.
  • the dedifferentiated plant protoplasts are dissolved in lower alcohols (C1 to C4 alcohols), mixed with the active substance and water, and sodium chloride is added, the active substance is introduced into the protoplasm by osmosis. It is pulled in.
  • lower alcohols C1 to C4 alcohols
  • This process is carried out in a pressurized osmosis process, specifically at 0.01 to 10 MPa, preferably at 0.05 to 1 MPa, most preferably at 0.05 to 0.5 MPa, wherein the concentration is 1% or more, preferably by addition of sodium chloride. It is set to 1-50% below. If the pressure and concentration are less than the above range, the introduction of the active substance does not occur effectively. If the pressure and the concentration are above the above range, the protoplasm may be destroyed. Therefore, it is suitably used within the above range.
  • the pulling is carried out so that the concentration of the active substance can be sufficiently exhibited, which may vary depending on the active substance, but has a pulling rate of 0.001 to 20% (based on weight%).
  • concentration is 0.001 to 15% by weight, preferably 0.01 to 10% by weight, and more preferably 0.1 to 10% by weight.
  • bisretin amidomethylpentane the active substance of Example 2
  • the concentration may be 10 to 5000 ppm, preferably 50 to 5000 ppm, and more preferably 100 to 500 ppm.
  • the pulling may further perform a dehydration reaction to increase the pulling rate.
  • step (f) the post-treatment obtains a structure in which the active material is introduced into the protoplasm through the post-treatment.
  • This aftertreatment is a conventional aftertreatment process in which unreacted material (active material not taken in) and salts are removed by centrifugation and washing.
  • the structure in which the active material is introduced into the protoplasm obtained through the above steps is maintained with the stability of the active ingredient existing in the plant cells as it is, and further, the stability of the active material introduced into the cells is high. Efficacy is also improved and can be preferably used as an active ingredient composition of the cosmetic.
  • Such structures comprise from 0.001 to 99.0% by weight, preferably from 0.01 to 10.0% by weight, more preferably from 0.1 to 3% by weight, based on the total weight of the total cosmetic composition.
  • the cosmetic composition may be prepared in any formulation as known, and may be a lotion, nourishing lotion, nourishing cream, massage cream, essence, pack, paste, gel, cream, lotion, powder, soap, oil, foundation, wax or It may be formulated in a spray form.
  • compositions of each formulation may contain various bases and additives necessary and appropriate for the formulation of the formulation, and nonionic surfactants, silicone polymers, extender pigments, fragrances, preservatives, Fungicides, oxidation stabilizers, organic solvents, ionic or nonionic thickeners, softeners, antioxidants, free radical destroying agents, opacifying agents, stabilizers, emollients, silicones, ⁇ -hydroxy acids, antifoams, humectants, And known compounds such as vitamins, insect repellents, fragrances, preservatives, surfactants, anti-inflammatory agents, substance P antagonists, fillers, polymers, propellants, basicizing or acidifying agents, or coloring agents.
  • the formulation of the present invention is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components.
  • animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components.
  • animal oils vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide
  • cellulose derivatives polyethylene glycols
  • silicones bentonites
  • silicas talc or zinc oxide
  • lactose 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, and especially in the case of spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.
  • a solvent, solubilizer or emulsifier 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.
  • liquid carrier diluents such as water, ethanol or propylene glycol
  • suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Soluble cellulose, aluminum metahydroxide, bentonite, agar or tracant and the like can be used.
  • the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, isethionate, an imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide.
  • Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.
  • Tissue sections (at least 3 cm) were cut from curry plants ( Helichrysum italicum ), gugal ( Commiphora wightii, Commiphora myrrha), stems and leaves of Opuntia Ficus indica . In this operating step, all work was performed on a sterile workbench under sterile conditions.
  • the tissues are soaked in 70% ethanol (Ethanol, Sigma, USA) for 60 seconds and 30% hydrogen peroxide (LG Chemical, Korea) for 15 minutes and the solvent is removed. Rinse 3 to 5 times with 2 O, soak for 15 minutes in sodium chlorate (Sigma, USA) to which a few drops of Tween 20 was added, and wash 3 to 5 times with sterile H 2 O.
  • tissue culture For tissue culture, these tissue fragments are placed in a sterile Petri dish (125 mm) and cut (2-3 mm) and carefully removed the bleached portion, and the sample thus obtained is thinly cut to give a solid culture medium (see table below). 1) smeared and half buried.
  • Two to three cell clusters (1 to 2 cm) were collected with a spatula, and then plated and dispersed in fresh medium. All this was done on a sterile workbench under sterile conditions.
  • the dedifferentiated plant cells were transferred to the liquid culture medium of Table 2 below, and then cultured in a rotary stirrer at 50 ° C. to 150 rpm under 25 ° C. and dark conditions, and their passage culture period was fixed every 10 days.
  • a multicomponent enzyme mixture (cellulase 1%, hemicellulase 2% and pectinase 0.5%) was added to the liquid culture medium in which the dedifferentiated plant cells were grown, and reacted at a rate of 50 rpm for 20 hours at a temperature range of 25 ° C. The cell wall was removed.
  • the protoplasts in the culture were obtained by centrifugation for 15 minutes under 200xg, and further purified by centrifugation for 15 minutes under 5,000xg.
  • centrifugation was performed at 5,000xg for 20 minutes using a centrifuge (Supra 22K, Hanil, Korea) to obtain a structure in which cyclohexanediol bisethylhexanoate was introduced into the protoplasm.
  • Example 2 The same procedure as in Example 1, except that betarapacyan ( ⁇ -Lapachone, Sigma, USA) instead of cyclohexanediol bisethylhexanoate as the active material, the beta-parapachon is introduced into the prototype Was obtained (structure / glycerine, 20/80 wt%).
  • betarapacyan ⁇ -Lapachone, Sigma, USA
  • Example 2 In the same manner as in Example 1, using the growth factor complex peptide composition of the following composition instead of cyclohexanediol bisethylhexanoate as the active material, to obtain a structure in which the growth factor complex peptide is introduced into the prototype (structure / Glycerin, 20/80 wt%).
  • Growth factor complex peptide composition is oligopeptide-34 (Oligopeptide-34, Caregen, Korea), oligopeptide-24 (Oligopeptide-24, Caregen, Korea), decapeptide-4 (Caregen, Korea), acetyl deca Peptide-3 (Acetyl Decapeptide-3, Caregen, Korea) and rh-polypeptide-4 (rh-Polypeptide-4, Bio-FD & C, Korea) were added to tertiary distilled water at 4,000 ppm (20,000 ppm total) for 30 minutes. Stirred ones were used.
  • Figure 2 (a) is a standard sample of cyclohexanediol bisethylhexanoate, (b) is a GC spectrum of the structure in which cyclohexanediol bisethylhexanoate is introduced into the prototype of Example 1, It can be seen that cyclohexanediol bisethylhexanoate as an active substance is introduced into the cells.
  • Figure 3 (a) is a standard sample bisretin amido methyl pentane, (b) is a GC spectrum of the structure in which the bisretin amido methyl pentane is introduced into the propoflast of Example 2, Bis as an active substance in the cell It can be seen that retinamido also introduces methylpentane.
  • Figure 4 is a beta-rappachon as a standard sample
  • (b) is a GC spectrum of the structure in which beta-rappachon is introduced into the propoflast of Example 3, the beta-parapachon as an active substance is introduced into the cell It can be seen that.
  • Cells into which the growth factor complex peptide of Example 4 was introduced were made through a matrix assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF mass spectrometry).
  • MALDI-TOF mass spectrometry matrix assisted laser desorption / ionization time-of-flight mass spectrometry
  • Figure 5 (a) is a growth factor complex peptide of the standard sample, (b) is a MALDI-TOF mass spectrometry graph of the structure in which the growth factor complex peptide is inserted in the propoflast of Example 4, the active material in the cell growth It can be seen that the factor complex peptide is introduced.
  • Cyclohexanediol bisethylhexanoate has a whitening effect and was tested using melanocytes to compare the whitening effect with the construct prepared in Example 1.
  • B16 cell line (ATCC CRL-6323), a rodent melanoma cell line, was treated with 10% FBS (Gibco Co), 100 ⁇ g / ml streptomycin, and 100 U / ml penicillin at 37 ° C. and 5% CO 2 .
  • FBS Gibco Co
  • streptomycin 100 ⁇ g / ml streptomycin
  • U / ml penicillin 100 ⁇ g / ml penicillin at 37 ° C. and 5% CO 2 .
  • the added DMEM Dulbecco's modified Eagle medium was incubated (Incubator; Thermo-scientific, USA).
  • a 96-well plate 1 ⁇ 10 5 per well was inoculated using medium DMEM, and the structure prepared in Example 1 and cyclohexanediolbisethylhexanoate and alpha-bisabolol, which are known to have a whitening effect, were prepared. Each concentration was made to treat 2ml of the sample solution. Since 1ppm of ⁇ -Melanocyte stimulating hormone ( ⁇ -MSH) was treated and incubated for 72 hours. Next, the medium was washed with phosphate buffer saline (PBS), and then 100 ⁇ l of melanin extract solution (1N NaOH + 50% DMSO) was added per well to lyse the cells at 80 ° C., followed by ELISA reader. The absorbance of melanin was measured at 492 nm.
  • PBS phosphate buffer saline
  • Mm amount of melanin in ⁇ -MSH treatment group
  • Figure 6 is a graph showing the effect of inhibiting the production of melanin according to each concentration of the structure of Example 1, cyclohexanediol bisethylhexanoate and alpha-bisabolol.
  • Example 6 in the case of the structure in which cyclohexanediol bisethylhexanoate was introduced into the protoplasm prepared in Example 1 according to the present invention, the melamine inhibitory effect was shown to be cyclohexanediol bisethylhexanoate and alpha- It was superior to bisabolol, and the higher the concentration, the greater the effect of inhibiting melanogenesis.
  • Bisretinamido methylpentane has a skin regeneration effect, and compared the collagen biosynthesis effect with the structure prepared in Example 2, wherein retinol and adenosine were further used as a comparative example.
  • Human dermal fibroblasts (Human dermal fibroblast, Gibco, USA) were seeded in each well of a 96-well microplate to be 1 ⁇ 10 4 cells for 100 U / ml penicillin, 100 ⁇ g / ml Cultured in 10% Dulbecco's Modified Eagle's Medium (DMEM) supplemented with streptomycin. The medium was maintained until 80% confluence of attached cells and passaged at 80% confluence. Medium was changed every two days until confluence.
  • DMEM Dulbecco's Modified Eagle's Medium
  • the subculture was washed with a phosphate buffered saline (PBS) in which the culture medium was removed, and the cells were dropped with 0.25% trypsin-EDTA (Gibco BRL, Grand Island, NY, USA), followed by centrifugation of the cell suspension. After that, the cell number was measured and subcultured three times again in the culture.
  • PBS phosphate buffered saline
  • Human normal fibroblasts are inoculated in 48-well microplates containing DMEM medium containing 10% FBS at a concentration of 1 ⁇ 10 5 cells / well and a CO 2 incubator at 5% concentration (CO 2 ). Incubated for 24 hours at 37 °C.
  • Example 2 bisretinamido methylpentane, retinol, and adenosine were replaced with serum-free DMEM medium adjusted for each concentration, and further cultured for 48 hours. Ascorbic acid 50 ⁇ g / ml is added to promote collagen synthesis before the last 24 hours of culture. After incubation, each well was washed, replaced with DMEM medium containing no serum, and further incubated for 24 hours.
  • PICP procollagen type I-peptide
  • Figure 7 is a graph showing the collagen synthesis capacity of the structure of Example 2, bisretinamido methylpentane, retinol and adenosine, the collagen synthesis ability of the active substance incoming cells of Example 2 according to the present invention is very excellent, the concentration is Higher results show that the synthesis performance is further improved.
  • Figure 8 is a graph showing the collagen synthesis ability of the structure, beta-rappachon, retinol and adenosine of Example 3, the collagen synthesis ability of the active material incoming cells of Example 3 according to the present invention is very excellent, the higher the concentration The synthesis performance is further improved.
  • Figure 9 is a graph showing the collagen synthesis ability of the structure, growth factor complex peptide, retinol and adenosine of Example 4, the collagen synthesis ability of the active material incoming cells of Example 3 according to the present invention is very excellent, the higher the concentration It shows that the synthesis ability is further improved.
  • sample treatment is done the same for D2, D5, D6 and D8)-Concentration of sample: 0.2%
  • Sample D0 three control batch skin tissues, and sample three skin tissues, D6 and D9, respectively.
  • Example 10 is an image showing epidermal regeneration before and after treatment of the structure of Example 4 for human skin tissue, it can be seen that the regeneration effect is very excellent.
  • Figure 11 is an image showing the degree of increase in glycosaminoglycans before and after treatment of the structure of Example 4 for human skin tissue
  • Figure 12 is an image showing the degree of increase of type I collagen, both of the present invention In accordance with the growth factor complex peptide in the protoplasm can be seen that the effect is excellent.
  • Example 4 Ingredient Content (% by weight) Structure of Example 4 0.5 Lipophilic monostearate 2.0 Cetearyl Alcohol 2.0 Stearic acid 1.5 Polysorbate 60 1.5 Sorbitan stearate 0.6 Hydrogenated Polyisobutene 1.0 Squalane 3.0 Mineral oil 5.0 Cyclomethicone 5.0 Dimethicone 1.0 Tocopherol Acetate 0.5 glycerin 5.0 Betaine 3.0 Triethanolamine 1.0 Xanthan Gum 0.05 incense Quantity antiseptic Quantity Pigment Quantity Distilled water Remaining amount Sum 100.00
  • a flexible lotion was prepared using a known method to have a composition as shown in Table 5 below.
  • Example 5 ingredient Content (% by weight) Structure of Example 4 0.5 glycerin 5.0 1,3-butylene glycol 3.0 Betaine 1.0 Allantoin 0.1 DL-panthenol 0.3 EDTA-2Na 0.02 Sodium hyaluronate powder 0.05 ethanol 5.0 Octyldodeceth-16 0.2 Polyoxyethylene Cured Castor Oil 0.2 incense Quantity antiseptic Quantity Pigment Quantity Purified water Remaining amount Sum 100.00
  • Example 6 ingredient Content (% by weight) Structure of Example 3 0.5 Glyceryl Stearate SE 1.5 Cetearyl Alcohol 1.0 Shea Butter 1.5 Polysorbate 60 1.3 Sorbitan stearate 0.5 Cured Vegetable Oil 1.0 Mineral oil 5.0 Squalane 3.0 Cyclomethicone 2.0 Dimethicone 0.8 Tocopherol Acetate 0.5 Carbomer 0.12 glycerin 5.0 1,3-butylene glycol 3.0 Sodium hyaluronate powder 0.05 Triethanolamine 0.12 incense Quantity antiseptic Quantity Pigment Quantity Distilled water Remaining amount Sum 100.00
  • Example 3 0.5 Lipophilic Monostearic Acid Glycerin 1.5 Cetearyl Alcohol 1.5 Stearic acid 1.0 Polysorbate 60 1.5 Sorbitan stearate 0.6 Isostearyl Isosterate 5.0 Squalane 5.0 Mineral oil 35 Dimethicone 0.5 Hydroxyethyl cellulose 0.12 glycerin 6.0 1,3-butylene glycol 3.0 Triethanolamine 0.3 incense Quantity antiseptic Quantity Pigment Quantity Distilled water Remaining amount Sum 100.00
  • Example 8 ingredient Content (% by weight) Structure of Example 2 0.5 glycerin 6.0 Betaine 5.0 PEG 1500 2.0 Allantoin 0.1 DL-panthenol 0.3 EDTA-2Na 0.02 Hydrogenated Lecithin 0.6 Hydroxyethyl cellulose 0.1 Sodium hyaluronate powder 0.08 Carboxy Vinyl Polymer 0.2 Triethanolamine 0.2 Ceramide 0.2 Octyldodecanol 3.0 Squalane 3.0 Polysorbate 60 0.4 Glyceryl Stearate SE 1.5 incense Quantity antiseptic Quantity Pigment Quantity Distilled water Remaining amount Sum 100.00
  • Example 9 ingredient Content (% by weight) Structure of Example 1 0.5 Polyvinyl alcohol 15 Cellulose gum 0.15 glycerin 3.0 PEG 1500 2.0 Betaine 2.0 DL-panthenol 0.4 Allantoin 0.1 Triethanolamine 0.2 Nicotinamide 0.5 ethanol 6.0 PEG 40 Cured Castor Oil 0.3 incense Quantity antiseptic Quantity Pigment Quantity Distilled water Remaining amount Sum 100.00

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Abstract

La présente invention concerne une structure dans laquelle une matière active est introduite dans un protoplaste végétal dé-différencié, son procédé de préparation et une composition cosmétique la contenant et, plus précisément, une structure dans laquelle une matière active est introduite dans un protoplaste végétal dé-différencié, son procédé de préparation et une composition cosmétique la contenant en tant que principe actif. Dans la structure, la stabilité des principes actifs compris dans des cellules de plantes existantes est conservée intacte, et la stabilité de la matière active introduite dans les cellules est élevée. La composition cosmétique contenant la structure peut garantir divers effets par l'intermédiaire des effets améliorés de la matière active.
PCT/KR2014/010012 2013-12-27 2014-10-23 Structure dans laquelle une matière active est introduite dans un protoplaste végétal dé-différencié, son procédé de préparation et composition cosmétique la contenant Ceased WO2015099274A1 (fr)

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CN116426457A (zh) * 2023-04-04 2023-07-14 山东爱维德生物科技有限公司 一种木立芦荟原生质体的提取工艺
CN116570547A (zh) * 2022-04-22 2023-08-11 华熙生物科技股份有限公司 一种植物细胞活性物和由其组成的透皮吸收促进剂组合物及其应用

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CN115778891A (zh) * 2022-12-26 2023-03-14 新生活化妆品科技(上海)有限公司 一种化妆品原料及其制备方法与应用、以及化妆品
KR20240147618A (ko) * 2023-03-31 2024-10-08 주식회사 엘지생활건강 피부 노화 개선용 화장료 조성물
KR102644064B1 (ko) * 2023-05-17 2024-03-05 한윤섭 포도, 올리브잎 및 몰약 혼합추출물을 유효성분으로 함유하는 피부 주름 개선용 화장료 조성물
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CN116919877B (zh) * 2023-08-23 2025-08-12 广州凌志生物科技有限公司 一种具有保湿舒缓组合物及其制备方法与应用

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CN116426457B (zh) * 2023-04-04 2023-09-29 山东鲁台农业科技有限公司 一种木立芦荟原生质体的提取工艺

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