WO2019190200A1 - Composition cosmétique comprenant un extrait de milieu conditionné d'adipocyte humain et du pdrn - Google Patents

Composition cosmétique comprenant un extrait de milieu conditionné d'adipocyte humain et du pdrn Download PDF

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WO2019190200A1
WO2019190200A1 PCT/KR2019/003587 KR2019003587W WO2019190200A1 WO 2019190200 A1 WO2019190200 A1 WO 2019190200A1 KR 2019003587 W KR2019003587 W KR 2019003587W WO 2019190200 A1 WO2019190200 A1 WO 2019190200A1
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pdrn
cell culture
stem cell
skin
culture extract
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PCT/KR2019/003587
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English (en)
Korean (ko)
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석재식
구대학
이원용
박경원
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(주)웰메이드코엔
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Publication of WO2019190200A1 publication Critical patent/WO2019190200A1/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
    • A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • A61K8/981—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of mammals or bird
    • 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
    • A61K8/60—Sugars; Derivatives thereof
    • 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
    • A61K8/60—Sugars; Derivatives thereof
    • A61K8/606—Nucleosides; Nucleotides; Nucleic acids
    • 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
    • A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • 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
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q7/00—Preparations for affecting hair growth

Definitions

  • the present invention relates to a cosmetic composition containing human fat stem cell culture extract and PDn, specifically, for improving wrinkles, whitening, elasticity, moisturizing and hair growth containing human fat stem cell culture extract and PDn It relates to a cosmetic composition for promotion.
  • the greatest areas of interest in cosmetics are wrinkles, pigmentation, decreased elasticity, skin drying and hair loss or lack of gloss associated with aging.
  • the skin undergoes various changes through aging. First, the thickness of the epidermis, dermis and subcutaneous tissue, which are components of the skin, becomes thin and the ECM (Extracellular Matrix) component that gives elasticity to the skin is changed.
  • ECM Extracellular Matrix
  • the connective tissue fibers of the ECM include glue fibers (collagen fibers, collagen fibers), reticulated fibers, and elastic fibers (elastic fibers, elastic fibers), which account for about 70% of skin connective tissue.
  • Collagen in most forms in the fibroblasts of the skin.
  • collagen content in skin connective tissues decreases due to a decrease in collagen synthesis and accelerated degradation. Therefore, degradation of collagen biosynthesis and promotion of collagen degradation may be the main cause of skin wrinkle formation.
  • Collagen biosynthesis processes are regulated by many factors involved in transcription and post-translation levels, and collagen degradation is collagenase, which breaks down collagen by UV light.
  • MMP matrix metalloproteases
  • collagen degradation is promoted to reduce the collagen content.
  • the skin becomes wrinkled and deepened.
  • skin aging is manifested by cell homogenization, loss of elastin, destruction of collagen, reduction and delay in collagen synthesis.
  • skin aging occurs in the skin epidermis, but rather in the dermis.
  • glycosaminoglycans a glycoprotein that contains hyaluronic acid, which is essential for maintaining skin moisture, is also active, giving skin elasticity, softness, flexibility and moisturizing properties.
  • vitamin C L-ascorbic acid
  • aha ( ⁇ -Hydroxy acid: AHA) containing cosmetic composition that is effective in exfoliation has also been widely used to improve the skin by promoting the activity of the cells with the exfoliation.
  • Stem cells are undifferentiated cells between differentiated cells of tissues or organs. They have self-renewing ability that continuously proliferates itself and have the ability to differentiate into all tissue cells in the body (Differentiation to specific tissue). Pluripotent cells.
  • hematopoietic stem cells include hematopoietic stem cells, mesenchymal stem cells, neuroblasts, epithelial hair cells, etc., and hematopoietic stem cells are capable of producing blood cells and lymphocytes, and are useful for treating immune system diseases including blood cancer.
  • hematopoietic stem cells In case of differentiation into bone, cartilage, adipose and fibrous tissues are involved in the recovery of each tissue is damaged.
  • these adult stem cells are present in small amounts in their tissues and may remain silent for years without dividing or proliferating. That is, stem cells are undifferentiated cells which have such differentiation ability but have not yet differentiated. Under these undifferentiated conditions, if appropriate conditions are met, stem cells can differentiate into various tissue cells. Therefore, the research for applying to the treatment, such as regeneration of damaged tissues, by using the differentiation capacity of stem cells are in progress.
  • Stem cells are also largely divided into embryonic stem cells and adult stem cells. Stem cells can be divided into two types: adult stem cells, which are present in various tissues in our bodies since birth, and embryonic stem cells, which are derived from fertilized eggs, which are the beginnings of human life. .
  • Adult stem cells are cells that constitute specific tissues, that is, bone marrow cells are blood cells, skin stem cells are skin, and neurons are destined to differentiate only olfactory nerve cells. Which cells differentiate into stem cells depends on the environment in which they are cultured. Therefore, if you create an environment to differentiate into neurons becomes a neuron, and if you create an environment to differentiate into skin cells can be a skin cell. In the present invention, it is intended to apply the stem cell culture extract to the cosmetic composition using these properties.
  • Stem cell cultivation method for producing human adipocyte conditioned media extracts is cultured for several generations of fat-depleted cells from adipose tissue to separate stem cells and then stem from stem cell culture.
  • the protein produced by the cell can be obtained.
  • This protein mixture contains EGF (epidermal growth factor), vEGF (vascular endothelial growth factor), TGF (transformation growth factor), HGF (hepatocyte growth factor), FGF (fibroblast growth factor), and IGF (insulin-like growth factor). It is reported that it contains about 150 growth factor protein components.
  • Adipose stem cells are a type of adult stem cells that are easier to harvest than bone marrow stem cells or umbilical cord blood stem cells. The growth factor is the most important factor in the proliferation and differentiation of one cell into another.
  • growth factors are responsible for transmitting signals that cause cells to proliferate or differentiate into cells, and based on these signals, cells divide and differentiate to make the skin fresh and lively, and always keep the skin fresh and fresh. You can keep it in the best condition.
  • the proteins produced from stem cells were applied to fibroblasts of the skin, the amount of collagen, a connective tissue produced by fibroblasts, increased more than five times, and the growth of fibroblasts increased by more than 30%.
  • Fibroblasts are cells that produce collagen, which forms the dermal layer of the skin, and when the amount of collagen decreases, skin elasticity decreases and wrinkles increase.
  • Stem cells are multipotent cells that are multipotent and have the ability to differentiate into cells such as bone, cartilage, ligaments, fat, myocardium, and muscle in vitro as well as in vivo. These multipotent cells, stem cells, are not only used as a good model in the study of differentiation mechanisms of individual tissue cells, but are also used in the development of various cell therapies.
  • mesenchymal stem cells In view of its application as a cell for cell treatment, mesenchymal stem cells have a number of advantages over embryonic stem cells in that cell harvesting is relatively easy, safe, and self-transplantable. Because of the advantages mentioned above, the treatment of various diseases such as myocardial infarction, fracture, cartilage damage, and stroke using stem cells is being studied.
  • stem cells are stem cells that can be utilized in culture in vitro, and have a very short lifespan and have a differentiation capacity for a short period of time. The limited lifespan of these stem cells is a major limitation in the utilization of the cells.
  • stem cells in the cosmetic field is the use of growth factors as metabolites.
  • Growth factors are proteins that bind to receptors on the cell's surface and stimulate cell proliferation and differentiation. Functions of growth factors are so diverse that they stimulate the division of many cells, while in some cases only specific cell types. Growth factors have been shown to regulate cell differentiation, migration and proliferation as essential components for stimulation of cell proliferation. Binding of growth factors to specific receptors activates signal transduction chains, which transmit signals from the outside of the cell to the inner nucleus. As signal transduction continues, it acts like a large amount of other transporters, finally synthesizing new proteins. Thus, growth factors form important signal transduction networks in the transmission of intercellular interactions and in the control of function.
  • PDRN Polydeoxyribonucleotide
  • PDRN is a nucleotide fragment of a specific specification, which is sodium DNA extracted from salmon, a regression fish.
  • PDRN which is already used as an injectable drug and has proven its stability and efficacy, has tissue regeneration efficacy and stimulates A2 receptor, which is a skin regeneration signal, to promote the secretion of various growth factors, Vascular endothelial growth factor (VEGF) )
  • VEGF Vascular endothelial growth factor
  • VEGF Vascular endothelial growth factor
  • PDRN is always present in human cells and physiologically stimulates the regeneration and metabolic activity of fibroblasts to help maintain the best condition.
  • PDRN Polydeoxyribonucleotide
  • PDRN Polydeoxyribonucleotide
  • PDRN improves the blood circulation of the scalp and can observe newborn hair 4 to 6 weeks after the procedure. The patient's satisfaction is high.
  • the use of PDRN in hair transplantation compensates for the disadvantages of conventional PRP treatment. And edema that occurs after transplantation.
  • PDRN regenerates and grows fibroblasts, which are skin stem cells, creates elastic fibers such as collagen and elastin, regenerates damaged skin quickly, and improves blood circulation to keep skin more elastic. It was judged to be a very effective way to maximize the effects of stem cells when used with various skin stem cells.
  • the cosmetic composition comprising the culture extract of the human adipose stem cells and PDRN (Polydeoxyribonucleotide; PDRN) as an active ingredient has an excellent wrinkle improvement, whitening effect, skin elasticity enhancing effect, moisturizing effect and hair growth promoting effect.
  • PDRN Polydeoxyribonucleotide
  • Korean Patent No. 10-1627562 discloses a cosmetic composition containing human adipose stem cell culture extract and hyaluronic acid.
  • Korean Patent Publication No. 10-2017-0068857 discloses a composition for anti-inflammatory and skin regeneration using polydeoxyribonucleotide.
  • An object of the present invention is to provide a cosmetic composition containing human adipose stem cell culture extract and PDRN, specifically, for improving wrinkles containing human adipose stem cell culture extract and PDRN.
  • a cosmetic composition for whitening, skin elasticity promotion, moisturizing and hair growth promoting.
  • the present invention is a cosmetic composition for improving wrinkles containing human fat stem cell culture extract and PDRN
  • the cosmetic composition is 0.01 to 10% by weight of human fat stem cell culture extract and 0.01 to 10% by weight of the total weight of the cosmetic It comprises 2.0 wt% PDRN, and the weight ratio of human fat stem cell culture extract and PDRN is 1: 1 to 10: 1.
  • the present invention is a whitening cosmetic composition containing human fat stem cell culture extract and PDRN, the cosmetic composition is 0.01 to 10% by weight of human fat stem cell culture extract and 0.01 to 2.0 to the total weight of the cosmetic It comprises weight% PDRN, and the weight ratio of the human fat stem cell culture extract and PDRN is 1: 1 to 10: 1.
  • the present invention is a cosmetic composition for enhancing skin elasticity containing human fat stem cell culture extract and PDRN, the cosmetic composition is 0.01 to 10% by weight of human fat stem cell culture extract and 0.01 to the total weight of the cosmetic To 2.0 wt% PDRN, and the weight ratio of the human adipose stem cell culture extract and PDRN (PDRN) is 1: 1 to 10: 1.
  • the present invention is a moisturizing cosmetic composition containing human fat stem cell culture extract and PDRN, the cosmetic composition is 0.01 to 10% by weight of human fat stem cell culture extract and 0.01 to 2.0 to the total weight of the cosmetic It comprises weight% PDRN, and the weight ratio of the human fat stem cell culture extract and PDRN is 1: 1 to 10: 1.
  • the present invention is a hair growth promoting cosmetic composition containing human fat stem cell culture extract and PDRN
  • the cosmetic composition is a human fat stem cell culture extract of 0.01 to 10% by weight relative to the total weight of the cosmetic and 0.01 to 2.0 wt% of PDRN
  • the weight ratio of the human fat stem cell culture extract and PDRN of PDRN is 1: 1 to 10: 1.
  • the human fat stem cell culture extract includes a culture solution obtained by culturing fat stem cells and all extracts obtained by crushing the cultured fat stem cells.
  • the human adipose stem cell means a stem cell obtained from an adult except for embryonic stem cells, and includes all stem cells derived from the base of blood and tissue including bone marrow, re-blood, and peripheral blood, and the present invention is not limited thereto. no.
  • the active ingredients of the human adipose stem cell culture extract are mainly peptides and proteins that regulate cell growth, for example PDGF (platelet-derived growth factor), Basic FGF (basic fibroblast growth factor), KGF (keratin cell growth factor). ), TGF- ⁇ 1 (transformation growth factor), HGF (hepatocyte growth factor), VEGF (vascular endothelial growth factor), collagen, fibronectin, SOD (antioxidant enzyme) and the like.
  • PDGF platelet-derived growth factor
  • Basic FGF basic fibroblast growth factor
  • KGF keratin cell growth factor
  • TGF- ⁇ 1 transformation growth factor
  • HGF hepatocyte growth factor
  • VEGF vascular endothelial growth factor
  • collagen fibronectin
  • SOD antioxidant enzyme
  • the human stem cell culture extract contains the growth factors described above, skin cell constituent cells promote fibroblast migration, activation effect, skin regeneration and anti-aging effect, collagen synthesis increase, tyrosinase activity inhibition effect, melanin synthesis inhibition, hair Growth and contraction of hair follicle restoration can be expected.
  • Human fat stem cell culture extract of the present invention can be obtained by the method specified in US Pat. No. 6,153,432 (2000.11.28). First, adipose stem cells are separated from adipose tissue obtained through liposuction. It is suspended in the substrate medium and inoculated in a culture vessel and then incubated for 24 hours at 37 °C, CO 2 incubator, when the fat stem cells adhere to the bottom of the culture vessel, the substrate medium is removed and put into differentiation medium (DIFFERENTIATION MEDIA). After incubation for 3 days, adipocyte medium (adipocyte media) is added and differentiated into adipocytes.
  • adipocyte medium adipocyte media
  • the differentiation of adipose stem cells begins at the beginning of differentiation and begins to form small lipid droplets in the cytoplasm, gradually increasing the number of small lipid droplets, and gradually adding smaller lipid droplets to form large lipid droplets. Once fully matured, one lipid droplet occupies the entire cytoplasm and gradually increases in size during the maturation process.
  • the culture medium is serum-free DMEM medium, basic fibroblast growth factor (bFGF) and / or epidermal growth factor (EGF) 0.1 ⁇
  • the culture broth obtained by culturing in DMEM medium further comprising 100 ng / ml concentration was separated and purified and used as the adipose stem cell culture extract of the present invention.
  • PDRN of the present invention is an abbreviation of polydeoxyribonucleotide and is called PDRN because it shows pharmacological activity without transmitting genetic information unlike DNA fragments or general DNA.
  • This is pure PDRN, an active form of 95% or higher purity obtained from salmon (Oncorhynchus keta) or trout (Oncorhynchus mykiss) semen and subjected to high heat treatment.
  • the INCI name is Sodium DNA.
  • PDRN polydeoxyribonucleotide
  • PDRN is composed of DNA polymer of specific size and when it is administered in biological tissues, it not only promotes DNA synthesis but also promotes rapid regeneration against excessive inflammatory reaction or tissue production.
  • the rapid anti-inflammatory and tissue regeneration effect shortens the wound healing time, activates cell regeneration, contains collagen, activates the production of various proteins and promotes the differentiation of various cells.
  • PDRN used in the present invention is sodium diene extracted from the testis of salmon, the GC content (%) of about 40%, the molecular weight is 1.3 ⁇ 10 6 Da ( ⁇ 2,000 bp).
  • VEGF Vascular endothelial growth factor vascular endothelial cell growth factor
  • PDRN Polydeoxyribonucleotide
  • VEGF vascular endothelial growth factor
  • various researches on the treatment of hair loss have been progressed and emerging as a new treatment method using PDRN such as hair follicle cell regeneration, scalp capillary generation, and scalp tissue regeneration using PDRN.
  • PDRN improves the blood circulation of the scalp and can observe newborn hair 4 to 6 weeks after the procedure. The patient's satisfaction is high. Also, the use of PDRN in hair transplantation compensates for the disadvantages of conventional PRP treatment.
  • PDRN regenerates and grows fibroblasts, which are skin stem cells, creates elastic fibers such as collagen and elastin, regenerates damaged skin quickly, and improves blood circulation to keep skin more elastic. It was judged to be a very effective way to maximize the effects of stem cells when used with various skin stem cells.
  • the cosmetic composition of the present invention may include 0.01 to 10.0% by weight of human fat stem cell culture extract and 0.01 to 2.0% by weight of PDRN based on the total weight of the cosmetic.
  • the weight ratio of human adipose stem cell culture extract and PDRN may be 1: 1 to 10: 1, preferably 1: 1 to 5: 1.
  • the cosmetic composition of the present invention containing human fat stem cell culture extract and PDRN as an active ingredient, skin wrinkle improvement effect, whitening effect, skin elasticity enhancing effect, moisturizing effect and hair growth promotion It has a skin anti-aging effect such as effect.
  • a cosmetic containing human fat stem cell culture extract and PDRN was prepared as in the following example.
  • composition containing sodium diene extracted from human adipose stem cell culture extract and PDRN testis, molecular weight of 1.3 ⁇ 10 6 Da ( ⁇ 2,000 bp) (Example 1)
  • a composition containing only human fat stem cell culture extract Comparative Example 1
  • a composition containing only PDRN Comparative Example 2
  • Example 1 In order to measure the skin wrinkle improvement effect (mechanical evaluation) for the cosmetic composition prepared by Example 1 according to the present invention, 30 women over 20 years old (mean age 45.5 years) were grouped into groups A, B, and C respectively. Divided into, the cosmetic composition prepared according to the embodiment of Table 1 according to the present invention was applied to Example 1 in Group A, Comparative Example 1 in Group B, Comparative Example 2 in Group C.
  • a replica of the skin wrinkles was applied to each subject of group A, B, and C by applying a transparent silicone solution while applying it to the area of the upper arm 2 ⁇ 2 cm 2 for 6 weeks (2 times / day, 0.2 g / time).
  • the skin wrinkles were measured using a skin wrinkle measuring instrument (SKIN VISIOMETER SV400, C + K Electronics GmbH. Germany).
  • SKIN VISIOMETER SV400 C + K Electronics GmbH. Germany.
  • Three-dimensional analysis of the image of the replica with a CCD camera wrinkles by the average wrinkle roughness (R z ) of the following formula 1 which is the sum of the roughness (R m : m is an integer of 1 or more) of each wrinkle divided by the number of wrinkles
  • the improvement effect (6 weeks) was analyzed.
  • Table 2 The test results are shown in Table 2 below.
  • Average wrinkle roughness (R z ) [R 1 + R 2 + R 3 +... + R m -2 + R m -1 + R m ] / number of wrinkles (m)
  • Example 1 As can be seen in Table 2, after six weeks, the average wrinkle roughness (R z ) of Example 1 according to the present invention was reduced by 93.7 ⁇ m (p ⁇ 0.01), Comparative Example 1 was 62.3 ⁇ m (p ⁇ 0.05), Comparative Example 2 is reduced by 52.3 ⁇ m (p ⁇ 0.05), the cosmetic containing a human body fat stem cell culture extract and PDRN according to the present invention is a comparative example containing only human body fat stem cell culture medium About 33.5% than 1, 44.2% than the comparative example 2 showed a wrinkle improvement effect.
  • the cosmetic composition containing human fat stem cell culture extract and PDRN according to the present invention is a human fat stem cell culture extract or PDAL & It can be seen that the effect of improving wrinkles is more effectively increased than the cosmetics containing (PDRN) alone.
  • each subject of group A, B, and C was classified into six grades for objective evaluation and subjective evaluation.
  • the improvement degree ( ⁇ W) of was measured.
  • Table 3 objective evaluation of the skilled person
  • Table 4 subjective evaluation of the subject.
  • Example 1 containing the human adipose stem cell culture extract and PDRN according to the present invention
  • the objective evaluation of the skilled person and the subjective evaluation of the subject were 2.7 and 2.6, respectively.
  • an increase of 33.3% and 44.4% of Comparative Example 2 was higher than that of Comparative Example 1, and in the subjective evaluation of the subject, it was found to be 26.9% higher than Comparative Example 1 and 38.5% higher than Comparative Example 2.
  • the cosmetics containing the adipose stem cell culture extract and PDRN have more effective wrinkle improvement than the cosmetics containing only human adipose stem cell culture extract or PDRN.
  • Melanosite was used by purchasing a mouse-derived B-16 melanoma (ATCC CRL 6323) cell line.
  • Melanoma cell lines were inoculated in DMEM medium containing 4.5 g / L glucose, 10% serum and 1% antibiotics and incubated at 37 ° C. in a 50 ml T-flask. After 24 hours of incubation under 5% CO 2 , the cells were isolated by treatment with 0.05% Trypsin containing 0.02% EDTA, and then incubated in a 50 ml T-flask for 48 hours. At this time the cell number is 5.76 ⁇ 10 6 cells / flask.
  • the human adipose stem cell culture extract and PDRN of the present invention were diluted in DMEM medium at 1, 2, 5, 10, 20, 50, and 100 ⁇ g / ml, respectively, and treated with cultured melanoma cells 37 Incubate for 5 days at °C. After incubation, all of the medium was removed, cells were treated with 1 ml of saline-phosphate buffer (PBS) containing 0.02% EDTA and 0.05% trypsin, and then cells were collected by centrifugation for 5 minutes. 5% trichloroacetate (TCA) is treated, stirred and centrifuged to wash the precipitated melanin with saline-phosphate buffer. 1N NaOH was dissolved in the melanin precipitated, and then the absorbance was measured at 475 nm. Melanin concentrations were determined from standard concentration curves of synthetic melanin (Sigma). The experimental results are shown in Table 5.
  • PBS saline-phosphate buffer
  • TCA t
  • the human fat stem cell culture extract of Example 1 according to the present invention and PDRN mixed with a weight ratio of 5: 1 human fat stem cells of Comparative Example 1 The melanin synthesis inhibitory effect was higher than the treatment with only the culture extract and the treatment with PDRN of Comparative Example 2.
  • the human fat stem cell culture extract and PDRN of the present invention are used together, the melanin production of melanocytes is more effective than when the human fat stem cell culture extract or PDRN is used alone. It can be seen that the skin whitening effect is suppressed.
  • Cosmetic Example 1 containing the human fat stem cell culture extract and PDRN of the present invention and the cosmetic containing only human fat stem cell culture extract of Comparative Example 1, which is a cosmetic containing only PDRN
  • the skin elasticity improvement effect of the comparative example 2 was measured.
  • Example 1 Comparative Example 1 Comparative Example 2 Subject 1 0.62 0.40 0.49 Subject 2 0.35 0.43 0.47 Subject 3 0.58 0.37 0.35 Subject 4 0.67 0.32 0.58 Subject 5 0.45 0.37 0.39 Subject 6 0.49 0.28 0.33 Subject 7 0.61 0.26 0.39 Subject 8 0.57 0.32 0.31 Subject 9 0.41 0.29 0.30 Subject 10 0.49 0.39 0.47 ⁇ R 0.52 0.34 0.41
  • Example 1 which is a cosmetic containing human fat stem cell culture extract and PDRN, has a skin elasticity of 34.6%, compared to Comparative Example 1 containing only human fat stem cell culture extract. Compared with Comparative Example 2 containing only PDRN, it increased by 21.2% compared to cosmetics containing only human fat stem cell culture extract and PDRN.
  • Cosmetic Example 1 containing only the human adipose stem cell culture extract and PDRN of the present invention and cosmetics containing only the human adipose stem cell culture extract extract Comparative Example 1 and cosmetics containing only PDRN The skin moisturizing effect for Example 2 was measured. The measurement was performed after stabilizing the skin for 20 to 30 minutes in a room having a temperature of 24-26 ° C., a relative humidity of 45%, and no air flow.
  • the cosmetic composition prepared according to Example 1 according to the present invention was divided into 30 groups (average age 45.5 years) of women over 20 years old, respectively, in Group A, the cosmetic composition prepared by Comparative Example 1
  • the cosmetic composition prepared by Comparative Example 2 in the silver B group was applied to the C group for 6 weeks (2 times / day) around the eyes, and then percutaneously using TEWAMETER TM210 (C + K electronic GmbH. Germany).
  • the amount of water loss, ie TEWL (Transepidermal Water Loss) is measured as the TEWL value change ⁇ TEWL over time, and the change in electrical conductivity according to the epidermal moisture content is determined using CORNEOMETER CM 820 PC (C + K electronic GmbH. Germany).
  • Moisturizing effect was measured by quantifying the moisture content of the skin from 0 to 150. The test results are shown in Table 7 below.
  • Example 1 As a result of the test, the ⁇ TEWL value of Example 1 was 6.8 g / hm 2, and 6 weeks later, the transdermal moisture loss was increased by 50.0% compared to Comparative Example 1 and 27.9% compared to Comparative Example 2, which is very effective. In addition, it can be seen that even in the moisture measurement value (Corneometer value) measuring the moisture content of the skin, Example 1 is increased by 31.7% compared to Comparative Example 1, 15.1% compared to Comparative Example 2, the skin is very moist.
  • composition containing only human fat stem cell culture extract and PDRN (PDRN) and the composition containing only human fat stem cell culture extract (Comparative Example 3) and PDRN and PDRN A composition containing (Comparative Example 4) was prepared.
  • Rat vibrissa immortalized dermal papilla cells (Filsell W, et al., 1994) were treated with 100 units / mL penicillin-100 ug / mL streptomycin (Gibco Inc, NY, USA) and 10heat-inactivated fetal bovin serum (FBS; Gibco Inc, NY). , USA) was used to culture in DMEM (Hyclone Inc, USA) medium at 37 °C, 5% CO 2 incubator, subcultured once every three days. Proliferation of rat vibrissa immortalized dermal papilla cells was measured using MTT assay.
  • Dermal papilla cells (1.0 ⁇ 10 4 cells / mL) were put in a 96well plate and incubated for 24 hours, exchanged with serum-free DMEM medium for 24 hours, followed by Example 2, Comparative Example 3, and Comparative Example 4 was treated at concentrations of 1, 10, 50 and 100 ⁇ g / ml respectively.
  • the supernatant is removed, dissolved in 200 ⁇ L of DMSO, and the absorbance is measured at 540 nm using a microplate reader (Amersh-am Pharmacia Biotech, NY, USA). The average absorbance values for each sample group were obtained, and the degree of proliferation was investigated by comparing with the absorbance values of the control group and shown in Table 9 below.
  • Example 2 was evaluated for the proliferative effect of dermal papilla dermal cells of Example 2 (composition containing human adipose stem cell culture extract and PDRN) of the dermal papilla It was found that the proliferative effect of the dermal cells is excellent. In particular, the proliferative effect of Dermal papilla cells of Example 2 showed a proliferative effect of dermal papillary dermal cells similar to the minoxidil sulfate, a positive control. Therefore, it was found that the human fat stem cell culture extract and PDRN used in Example 2 exhibited an excellent hair growth promoting effect on hair follicle growth.
  • 5 ⁇ -reductase activity was assessed by converting [3H] testosterone (T) into [3H] dihydrotestosterone (DHT) using rat prostate 5 ⁇ -reductase.
  • T testosterone
  • DHT dihydrotestosterone
  • Male 7 to 8 week old Wistar rats were anesthetized with ethyl ether, followed by cervical slaughter to remove the prostate, immediately stored in liquid nitrogen, and the required amount of prostate was taken out for each experiment.
  • the prostate was placed in a glass homogeniger containing 4 ° C homogeniger buffer (20 mM potassium phosphate, pH6.5, 0.32 M sucrose, 0.1 mM dithiothreitol (DTT)) and ground until it became cloudy.
  • Example 2 according to the present invention can be seen to exhibit a very good 5 ⁇ -reductase inhibitory effect.
  • the effect of inhibiting 5 ⁇ -reductase when the human adipose stem cell culture extract and PDRN were used together showed a 5 ⁇ -reductase inhibitory effect similar to that of minoxidil sulfate, a positive control. It can be seen that there is a promoting effect.
  • Example 1 The stability test of the formulation for the cosmetic containing the human adipose stem cell culture extract of the present invention and PDRN (PDRN) was performed.
  • Example 2 Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4 of the Table 1 in order to test the stability of the cosmetic constant temperature bath is kept constant at 45 °C
  • the degree of separation, discoloration and odor was measured and compared. The results are shown in Table 11 below. At this time, the product separation, discoloration, and degree of odor were evaluated by classifying into the following six grades.
  • Comparative Example 1 containing only the human adipose stem cell culture extract and PDRN of Example 1, Example 2 and human adipose stem cell culture extract of the present invention
  • Comparative Example 3 and Comparative Example 2 containing only PDRN and PDRN 4 it can be seen that there is no separation, discoloration and odor (specific odor) phenomenon is a stable base.
  • the patch is applied for 24 hours, and after the patch is removed, the test site is marked with a marking pen, and the test site is observed after 24 hours and 48 hours, respectively. Determination was performed 24 hours and 48 hours after patch removal, and skin reactions were determined according to the regulations of the International Contact Dermatitis Research Group (ICDRG) shown in Table 12 below. The test results are shown as skin stability in Table 13 below.
  • IDRG International Contact Dermatitis Research Group
  • Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 24 hours 0.0 0.0 0.0 0.0 0.0 0.0 48 hours 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0
  • Example 1 As shown in Table 13, as a result of skin safety test, in Example 1, Example 2, Comparative Example 1, Comparative Example 2, Comparative Example 3, and Comparative Example 4, the average irritation degree is 0, and there is no irritation on the skin. It can be seen that it is a safe cosmetic.
  • composition of the present invention is not intended to be limited to the following formulation examples.

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Abstract

L'objectif de la présente invention est de fournir une composition cosmétique comprenant un extrait de milieu conditionné d'adipocyte humain et du PDRN et, plus particulièrement, une composition cosmétique pour la réduction des rides, le blanchiment, l'amélioration de l'élasticité de La peau, l'hydratation et la stimulation de la pousse des cheveux, comprenant un extrait de milieu conditionné d'adipocytes humains et du PDRN.
PCT/KR2019/003587 2018-03-30 2019-03-27 Composition cosmétique comprenant un extrait de milieu conditionné d'adipocyte humain et du pdrn WO2019190200A1 (fr)

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KR10-2018-0037607 2018-03-30

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KR102260525B1 (ko) 2019-11-15 2021-06-03 주식회사 엑소코바이오 줄기세포 유래 엑소좀과 폴리데옥시리보뉴클레오티드를 유효성분으로 포함하는 새로운 조성물
WO2021096089A1 (fr) * 2019-11-15 2021-05-20 주식회사 엑소코바이오 Nouvelle composition comprenant des exosomes dérivés de cellules souches et du polydésoxyribonucléotide en tant que principes actifs
KR102478551B1 (ko) * 2022-06-30 2022-12-19 주식회사 지에프메디컬 인체 유래 성분을 이용한 화장료 조성물
KR102534797B1 (ko) 2022-10-24 2023-05-30 주식회사 에이바이오머티리얼즈 지모추출물, pdrn 및 항산화 활성을 갖는 수용성 생리활성물질을 포함하는 나노 입자의 제조방법 및 이를 함유하는 화장료 조성물
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