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WO2016047823A1 - Composition pour inhiber la sénescence cellulaire, contenant en tant que principe actif un extrait de melandrium firmum rohrbach ou du bornésitol qui en est séparé - Google Patents

Composition pour inhiber la sénescence cellulaire, contenant en tant que principe actif un extrait de melandrium firmum rohrbach ou du bornésitol qui en est séparé Download PDF

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Publication number
WO2016047823A1
WO2016047823A1 PCT/KR2014/008944 KR2014008944W WO2016047823A1 WO 2016047823 A1 WO2016047823 A1 WO 2016047823A1 KR 2014008944 W KR2014008944 W KR 2014008944W WO 2016047823 A1 WO2016047823 A1 WO 2016047823A1
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extract
aging
pharmaceutical composition
zgc
cells
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Korean (ko)
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김재룡
손종근
양효현
황보경
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Research Cooperation Foundation of Yeungnam University
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Research Cooperation Foundation of Yeungnam University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/36—Caryophyllaceae (Pink family), e.g. babysbreath or soapwort

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  • the present invention relates to a composition for inhibiting cellular aging containing Jangchaechae extract or (-)-bornenesitol ((-)-bornesitol) separated therefrom as an active ingredient.
  • telomeres at the ends of the chromosomes become shorter in the process of cell division, resulting in DNA damage, which is called replication aging.
  • Cell aging is induced not only by shortening of telomeres, but also by dysfunction of oncogenes and cancer suppressor genes, inflammatory reactions, oxidative stress, anticancer agents, ultraviolet rays and radiation. Aging cells are larger in size and flatter in shape, stop cell growth, have many signs of DNA damage in the nucleus, and secrete a variety of inflammatory proteins.
  • SA- ⁇ -gal senescence-associated ⁇ -galactosidase
  • Cell aging may also inhibit or promote cancer, and has been suggested as an important mechanism of tissue regeneration and repair, tissue / individual aging and aging-related diseases.
  • cell aging contributes to the pathogenesis of various aging-related diseases such as cancer, arteriosclerosis, skin aging, neurodegenerative diseases, myotropenia, osteoporosis and prostatic hyperplasia.
  • aging-related diseases such as cancer, arteriosclerosis, skin aging, neurodegenerative diseases, myotropenia, osteoporosis and prostatic hyperplasia.
  • Recent studies have shown that selective control of cell aging can regulate the development of tissues, organs, aging, health life, and age-related diseases.
  • Telomerase deficient mice are known to be rapidly aging, and it has been shown that increasing telomerase expression in older telomere deficient mice reverses degenerative changes in tissues or organs with aging.
  • Jangguchae is a biennial plant that has been widely distributed in Korea and used to treat various diseases. It is reported in the literature that it has been widely used in the treatment of urinary, breast cancer, gonorrhea and lactation diseases. Butanol fraction of methanol extract of Jangchaechae has been reported to cause hepatic activity including hexobarbital prolongation of sleep, elevated serum transaminase activity, and severe pathological histological changes of hepatic cells. The effect of inhibiting cell or fibroblast aging has not been reported yet.
  • Korean Patent Registration No. 10-0686260 relates to a pharmaceutical composition and health functional food for liver function improvement and liver disease treatment comprising at least one of jangguchae and Ulleungjangguchae extract as an active ingredient, jangguchae or Ulleung jangguchae extract Not only inhibits AST and ALT activity, but also significantly inhibits the amount of collagen, alpha-smooth-muscle actin and TGF- ⁇ accumulated due to fibrosis in liver tissue, which is useful for the prevention and treatment of liver disease. Although it is disclosed that it can be used as a pharmaceutical and health functional food, there is no mention of inhibiting aging of umbilical vein endothelial cells or fibroblasts as in the present invention.
  • the present invention provides a pharmaceutical composition for inhibiting cellular aging containing Jangchaechae extract or (-)-bornenesitol ((-)-bornesitol) represented by the following Chemical Formula 1 isolated therefrom as an active ingredient.
  • Jangchaechae extract or (-)-bornenesitol ((-)-bornesitol) isolated therefrom inhibits cellular senescence by adriamycin and also inhibits aging due to cell division.
  • it can be usefully used for treating diseases related to aging, for example, skin aging, rheumatoid arthritis, osteoarthritis, hepatitis, chronic skin damage tissue, arteriosclerosis, prostate hyperplasia and liver cancer.
  • diseases related to aging for example, skin aging, rheumatoid arthritis, osteoarthritis, hepatitis, chronic skin damage tissue, arteriosclerosis, prostate hyperplasia and liver cancer.
  • anti-aging functional foods, anti-vascular aging drugs, and cosmetics that can inhibit cell aging of vascular endothelial cells and fibroblasts are expected to be utilized.
  • Figure 1 shows an isolated schematic diagram of Jangchaechae extract.
  • Figure 2 shows the chemical structure of a single component separated from Jangguchae.
  • Figure 3 shows the cytotoxicity and inhibitory effect of cell aging by adriamycin of Jangchaechae extract in human umbilical vein endothelial cells.
  • A Cytotoxic effect of Jangchaechae extract. Each extract was treated with 10, 100 ug / ml, and then cultured for 3 days to examine the cytotoxic effect by MTT method.
  • B SA- ⁇ -gal active staining picture and SA- ⁇ -gal active staining percentage. After adriamycin treatment, each extract was treated with 10 ug / ml, and 3 days later, SA- ⁇ -gal activity staining was performed. The results were expressed as mean and standard deviation after each experiment was repeated three times or more independently. Cont, control; DMSO, dimethylsulfoxide; NAC, N-acetylcysteine; Rap, rapamycin. * p ⁇ 0.05 or ** p ⁇ 0.01 vs DMSO.
  • Figure 4 shows the cytotoxicity of the Jangchaechae ethyl acetate extract in the human umbilical vein endothelial cells and the inhibitory effect of cell aging by adriamycin.
  • A Cytotoxic effect of Jangchaechae ethyl acetate extract. Each extract was treated with 1-10ug / ml, and then cultured for 3 days to examine the cytotoxic effect by MTT method.
  • B and C SA- ⁇ -gal active staining picture and SA- ⁇ -gal active staining percentage.
  • Figure 5 shows the effect of inhibiting the replication cell aging of Jangchaechae ethyl acetate extract in human umbilical vein endothelial cells.
  • A Cytotoxic effect of Jangchaechae ethylacetate extract on cloned aging cells. Each extract was treated with 1-10ug / ml, and then cultured for 3 days to examine the cytotoxic effect by MTT method.
  • B and C SA- ⁇ -gal active staining picture and SA- ⁇ -gal active staining percentage. 1-10ug / ml of Jangchaechae ethyl acetate extract was applied to cloned aging cells, and 3 days later, SA- ⁇ -gal activity staining was performed.
  • Figure 6 shows the cytotoxicity of a single component of Jangguchae in human umbilical vein endothelial cells and the inhibitory effect of cell aging by adriamycin.
  • A Cytotoxic effect of jang guchae single component. Each component was incubated for 10 days after 10ug / ml treatment, and the cytotoxic effect was examined by MTT method.
  • B SA- ⁇ -gal active staining percentage. After adriamycin treatment, each single component was treated with 10 ug / ml, and 3 days later, SA- ⁇ -gal active staining was performed. The results were expressed as mean and standard deviation after each experiment was repeated three times or more independently. Cont, control; DMSO, dimethylsulfoxide; NAC, N-acetylcysteine. * p ⁇ 0.05 vs DMSO.
  • Figure 7 shows the cytotoxicity of the jangguchae single component in human umbilical vein vascular endothelial cells and cell aging inhibitory effect by adriamycin.
  • A Cytotoxic effect of jang guchae single component. Each component was treated with 1ug / ml, and then cultured for 3 days, and the cytotoxic effect was examined by MTT method.
  • B SA- ⁇ -gal active staining percentage. After adriamycin treatment, ZGC-3 was treated with 1 ug / ml, and 3 days later, SA- ⁇ -gal activity staining was performed. The results were expressed as mean and standard deviation after each experiment was repeated three times or more independently. Cont, control; DMSO, dimethylsulfoxide; NAC, N-acetylcysteine. ** p ⁇ 0.01 vs DMSO.
  • Figure 8 shows the cytotoxicity and inhibitory effect of cell aging by adriamycin of Jangchaechae extract in human fibroblasts.
  • A Cytotoxic effect of Jangchaechae extract. Each extract was treated with 10, 100 ug / ml, and then cultured for 3 days to examine the cytotoxic effect by MTT method.
  • B SA- ⁇ -gal active staining picture and SA- ⁇ -gal active staining percentage. After adriamycin treatment, each extract was treated with 10 ug / ml, and 3 days later, SA- ⁇ -gal activity staining was performed. The results were expressed as mean and standard deviation after each experiment was repeated three times or more independently. Cont, control; DMSO, dimethylsulfoxide; NAC, N-acetylcysteine; Rap, rapamycin. * p ⁇ 0.05 or ** p ⁇ 0.01 vs DMSO.
  • Figure 9 shows the cytotoxicity and inhibitory effect of cell aging by adriamycin of Jangchaechae hexane extract in human fibroblasts.
  • A showing the cytotoxic effect of jangguchae hexane extract. Each extract was treated with 1-10ug / ml, and then cultured for 3 days to examine the cytotoxic effect by MTT method.
  • B and C SA- ⁇ -gal active staining picture and SA- ⁇ -gal active staining percentage. After adriamycin treatment, Jangchaechae hexane extract was treated with 1-10 ug / ml, and 3 days later, SA- ⁇ -gal activity staining was performed.
  • Figure 10 shows the effect of inhibiting replication cell aging of Jangchaechae hexane extract in human fibroblasts.
  • A the cytotoxic effect of Jangchaechae hexane extract on cloned aging cells. Each extract was treated with 1-10ug / ml, and then cultured for 3 days to examine the cytotoxic effect by MTT method.
  • B and C SA- ⁇ -gal active staining picture and SA- ⁇ -gal active staining percentage.
  • the cloned aging cells were treated with Jangchaechae hexane extract 1-10 ug / ml, and 3 days later, SA- ⁇ -gal activity staining was performed.
  • Figure 11 shows the cytotoxicity of the Jangchaechae single component in human fibroblasts and the effect of inhibiting cell aging by adriamycin.
  • A Cytotoxic effect of jang guchae single component. Each component was incubated for 10 days after 10ug / ml treatment, and the cytotoxic effect was examined by MTT method.
  • B SA- ⁇ -gal active staining percentage. After adriamycin treatment, ZGC-5, ZGC-6, and ZGC-9 were treated at 10 ug / ml, and 3 days later, SA- ⁇ -gal active staining was performed. The results were expressed as mean and standard deviation after each experiment was repeated three times or more independently. Cont, control; DMSO, dimethylsulfoxide; NAC, N-acetylcysteine. * p ⁇ 0.05 vs DMSO.
  • Figure 12 shows the effect of inhibiting cell aging by adriamycin of Jangchaechae single component ZGC-9 ((-)-bornenesitol) in human fibroblasts A.
  • B SA- ⁇ -gal activity Staining Photos and Percentage of SA- ⁇ -gal Active Staining
  • ZGC-9 was treated at 1-10 ug / ml and SA- ⁇ -gal active staining was performed 3 days later.
  • FIG. 13 shows the effect of ZGC-9 ((-)-bornesitol) on free radical production in human fibroblasts.
  • A free radical flow cytometry.
  • B average free radicals comparison. Fibroblasts were treated with adriamycin and ZGC-9 was treated with 10 ug / ml, and the level of intracellular free radicals was examined by flow cytometry. Each experiment was repeated three times or more independently, and the mean and standard deviation were expressed.
  • ADR adriamycin; Y, young cells; Cont, control; DMSO, dimethylsulfoxide; NAC, N-acetylcysteine; Rap, rapamycin.
  • ZGC-9 ((-)-bornesitol) on replication senescence in human fibroblasts.
  • A SA- ⁇ -gal active staining pictures are shown.
  • B SA- ⁇ -gal active staining percentage.
  • C ZGC-9 shows the cytotoxic effect.
  • the cloned cells were treated with 1-10 ug / ml of ZGC-9 and subjected to SA- ⁇ -gal activity staining 3 days later. Cytotoxicity was examined by MTT method. The results were expressed as mean and standard deviation after each experiment was repeated three times or more independently. Old, Old cells; DMSO, dimethylsulfoxide; NAC, N-acetylcysteine; Rap, rapamycin.
  • Figure 15 shows the cytotoxicity of Jangchaechae single component ZGC-2 (ursolic acid) in human fibroblasts and the inhibition of cell aging by adriamycin.
  • A shows the cytotoxic effects of ZGC-2.
  • SA- ⁇ -gal activity staining pictures and SA- ⁇ -gal activity staining percentages After adriamycin treatment, ZGC-2 was treated with 1 ug / ml, and 3 days later, SA- ⁇ -gal activity staining was performed. Results were shown as mean and standard deviation after each experiment repeated three or more times: Cont, control; DMSO, dimethylsulfoxide; NAC, N-acetylcysteine; Rap, rapamycin (rapamycin). * p ⁇ 0.05 or ** p ⁇ 0.01 vs DMSO.
  • Figure 16 shows the effect of inhibiting cell aging by adriamycin of Jangchaechae single component ZGC-2 (ursolic acid) in human fibroblasts.
  • a and B SA- ⁇ -gal activity staining photos and SA- ⁇ -gal Percentage of Active Staining
  • ZGC-2 was treated with 0.01-1 ug / ml and SA- ⁇ -gal active staining was performed three days later. The mean and standard deviation are shown, followed by the expression of C, p53, phosphorylated S6K and p21
  • ZGC-2 was treated with 0.01-1 ug / ml, and the expression level of each protein was determined by Western blot.
  • NT untreated; Cont, control; DMSO, dimethylsulfoxide; NAC, N-acetylcysteine; Rap, rapamycin.
  • Figure 17 shows the effect of ZGC-2 (ursolic acid) on the production of free radicals in human fibroblasts.
  • A free radical flow cytometry.
  • B average free radicals comparison. Fibroblasts were treated with adriamycin, ZGC-2 was treated with 1 ug / ml, and the level of free radicals in the cells was examined by flow cytometry, and each experiment was repeated three or more times independently, indicating the mean and standard deviation ADR, adriamycin; Y, young cells; Cont, control; DMSO, dimethylsulfoxide; NAC, N-acetylcysteine; Rap, rapamycin. * P ⁇ 0.05 or ** p ⁇ 0.01 vs DMSO.
  • Figure 18 shows the effect of inhibiting replication cell aging of ZGC-2 (ursolic acid) in human fibroblasts.
  • A shows the SA- ⁇ -gal activity staining pictures.
  • B SA- ⁇ -gal activity staining percentage ZGC-2 was treated with 0.01-1 ug / ml of cloned aging cells and subjected to SA- ⁇ -gal activation staining 3 days later. The results were repeated three or more times independently of each experiment. Variation: Old, Old cells; DMSO, Dimethylsulfoxide; NAC, N-acetylcysteine; Rap, Rapamycin. ** p ⁇ 0.01 vs DMSO.
  • Jangchaechae ethyl acetate extract in human vascular endothelial cells and Jangchaechae hexane extract, ZGC-2 (ursolic acid) and ZGC-9 ((-)-bornesitol) in human fibroblasts were found to be effective in inhibiting cell aging.
  • the invention was completed.
  • the present invention provides a pharmaceutical composition for inhibiting cellular aging containing Jangchaechae extract or (-)-bornenesitol ((-)-bornesitol) represented by the following Chemical Formula 1 isolated therefrom as an active ingredient.
  • the jangguchae extract is hexane (n-hexane) fraction extract extracted by adding distilled water and hexane (n-hexane) to the jangguchae methanol extract and fractionated, more specifically, the cells are fibroblasts It is done.
  • the jangguchae extract is ethyl acetate (EtOAc) fraction extract extracted by adding ethyl acetate (EtOAc) to the distilled water layer fractionated by adding distilled water and hexane (n-hexane) to the methanol extract of Jangguchae More specifically, the cells are characterized in that the umbilical vein vascular endothelial cells.
  • the cellular senescence is characterized by induced by adriamycin, the cell aging inhibition is to measure the inhibition of senescence-associated ⁇ -galactosidase (SA- ⁇ -gal) activity It features.
  • the pharmaceutical composition may include a pharmaceutically acceptable carrier in addition to the jang-guchae extract or (-)-bornenesitol ((-)-bornesitol), such a pharmaceutically acceptable Carriers to be used are commonly used in pharmaceutical preparations, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrroly Don, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but are not limited thereto.
  • the pharmaceutical composition may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like as an additive.
  • the pharmaceutical composition is determined by the method of administration according to the degree of symptoms of cellular aging, usually topical administration is preferred.
  • the dosage of the active ingredient in the pharmaceutical composition may vary depending on the route of administration, the degree of the disease, the age, sex, and weight of the patient, and may be administered once to several times daily.
  • the pharmaceutical composition may be administered to various mammals such as rats, mice, livestock, humans, and the like. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.
  • compositions may be prepared in unit dose form or formulated using pharmaceutically acceptable carriers and / or excipients or may be prepared within a multi-dose container.
  • the formulation may be in the form of a solution, suspension, or emulsion, or may be in the form of an exercicide, extract, powder, granule, tablet, warning, lotion, ointment, or the like.
  • the pharmaceutical composition may treat any one disease selected from the group consisting of skin aging, rheumatoid arthritis, osteoarthritis, hepatitis, chronic skin damage tissue, arteriosclerosis, prostate hyperplasia and liver cancer, but is not limited thereto.
  • Fraction BZG12 (0.3 g) was loaded onto a 75 ⁇ 2 cm Sephadex TM LH-20 column to obtain ZGC-7 (13.5 mg) and ZGC-8 (50.5 mg) with a solvent composition of 100% MeOH.
  • DMEM Dubroccos-Modified Eagle's medium
  • Fetal Bovine Serum Fetal Bovine Serum
  • Antibiotic Solution Penicillin-Streptomycin WelGene (Daegu, Korea)
  • Endothelial Cell Growth Medium-2 -2, EGM-2) was purchased from Lonza (Walkersvill, MD, USA).
  • Antibodies to p53 are described in SantaCruz Biotech, Inc. (SantaCruz, CA, USA), and antibodies against p21 and pS6 were from Cell Signaling Technology Inc. (Beverly, MA, USA).
  • GAPDH antibody was distributed by Dr. Ki-sun Kwon, Korea Research Institute of Bioscience and Biotechnology. Adriamycin used products of Ildong Pharmaceutical Co., Ltd.
  • Human fibroblasts were cultured using a DMEM medium containing 10% fetal bovine serum and 1% antibiotics (penicillin 10,000 units / ml, streptomycin 10,000 ug / ml). After dispensing 10 5 , it was incubated in 37 °C, 5% carbon dioxide incubator. When the cells grew to 80-90% at the bottom of the culture dish, trypsin-EDTA solution (2.5X) was added to separate the cells, and then passaged. Umbilical cord vascular endothelial cells were cultured in the same manner as human fibroblasts using EGM-2 as a culture medium. Each time the cells were passaged, the cell number was measured to determine how many times the cells divide.
  • PD Population doubling
  • MTT method The effect of jangguchae extract and compound on the growth rate of cells was investigated by MTT method. 0.1% MTT solution was added to 50 ul of each well of a 96 well culture vessel, and the reaction was performed at 37 ° C. and a 5% carbon dioxide incubator for 3 hours. After removing the culture solution and the MTT solution, 100 ul of dimethyl sulfoxide was added to dissolve the crystals formed. The growth rate of the cells was measured by measuring the absorbance at 550 nm using a microplate reader.
  • SA- ⁇ -gal activity staining After treatment with a single compound in a 24 well or 12 well culture vessel for 3 days, the cells were washed with phosphate buffer. After fixing the cells with 3.7% paraformaldehyde (paraformaldehyde), the fixed solution was removed and washed again with phosphate buffer.
  • SA- ⁇ -gal staining solution [40 mM citric acid / phosphate; pH 5.8, 5 mM potassium ferrocyanide, 5 mM potassium ferricyanide, 150 mM NaCl, 2 mM MgCl 2 , X-gal 1 mg / ml] were added to each well in a 24 well culture vessel.
  • Each cell was dispensed into 1 ⁇ 10 5 in a 60 mm culture dish and then cultured in a 37 ° C., 5% carbon dioxide incubator. After washing the cells twice with DMEM broth containing antibiotics, Jangchaechae extract and single components were treated for 1 hour before concentration, and adriamycin 500 nM for 4 hours. After removing the culture solution, it was washed twice with phosphate buffer.
  • Cell lysis solution per culture dish [25 mM Tris-HCl (pH 7.6), 150 mM Nacl, 1% Tryton X-100, 0.5% sodium deoxycholate, 0.1% SDS, 1 mM sodium vanadate vanadate), 5 mM NaF, protease inhibitor or 1 mM PMSF] was added 50 ul.
  • the plate was scraped using a cell scraper to collect the solution and cells and transferred to the microacupuncture tube.
  • the solution was shaken every 10 minutes while reacting for 30 minutes on ice.
  • the supernatant was transferred to a new tube by centrifugation for 15 minutes at 12,000 rpm.
  • the amount of protein in the solution was quantified by bicinchoninic acid (BCA) method (Pierce Biotechnology Inc., Rockford IL, USA) using bovine serum albumin as a standard protein.
  • BCA bicinchoninic acid
  • Protein (30 ⁇ g) was isolated by electrophoresis on 10% SDS-polyacrylamide gel. After transferring the protein to the nitrocellulose membrane, it was reacted for 30 minutes in Tween-20-Tris buffered saline (TTBS) containing 5% whole milk powder. The nitrocellulose membrane was reacted overnight in a 5% whole milk powder-TTBS solution containing primary antibodies against p53, pS6 or p21. After washing three times with TTBS solution three times, and reacted with a horseradish peroxidase-bound secondary antibody for 1 hour 30 minutes. After washing the membrane five times for 7 minutes with TTBS, the amount of protein was measured using an enhanced chemiluminescence solution.
  • TTBS Tween-20-Tris buffered saline
  • the amount of specific protein reacted with each antibody was measured using a LAS-3000 imaging device (Fujifilm Corp., Stanford, CT, USA). The same amount of protein was used in each experiment and compared with glyceraldehyde-3-phosphate dehydrogease (GAPDH) antibody.
  • GPDH glyceraldehyde-3-phosphate dehydrogease
  • the cells were dispensed into 1.5 ⁇ 10 5 in a 100 mm culture dish and incubated for 3 days in a 37 ° C., 5% carbon dioxide incubator.
  • the cells were washed twice with DMEM broth containing antibiotics and then treated with adriamycin 500 nM for 4 hours.
  • the cells were washed once with phosphate buffer, treated with trypsin-EDTA solution (2.5%), and cells were separated, and then aliquoted into 1 ⁇ 10 5 in a 60 mm culture dish. Incubated at 37 ° C., 5% carbon dioxide incubator for one day.
  • the culture solution was changed and a single compound was treated with 10 ug / ml and 1 ug / ml.
  • Dimethyl sulfoxide was added as a negative control and 5 mM of N-acetylcysteine and 500 nM of rapamycin were added as a positive control. After incubation for 3 days at 37 °C, 5% carbon dioxide incubator, washed twice with DMEM culture medium containing antibiotics and was treated with H 2 DCFDA 250uM for 20 minutes. Once washed with phosphate buffer solution, trypsin-EDTA solution was added to the cells were separated and transferred to the microneedle tube.
  • Jangchaechae ethyl acetate extract has an effect of inhibiting replication aging due to cell aging and cell division by adriamycin.
  • the cytotoxicity of 12 single components of Jangchaechae in human umbilical vascular endothelial cells was investigated.
  • the compounds were treated with 10 ug / ml, all of the other substances except ZGC-2 and 3 were not cytotoxic (FIG. 6A).
  • ZGC-6, 7, 10, 11, 12 was investigated whether inhibiting cell aging by adriamycin, but the effect of inhibiting cell aging could not be observed (Fig. 6B).
  • the concentration of ZGC-2, 3, which was toxic at 10 ug / ml was lowered by 1 ug / ml, no cytotoxicity was observed (FIG. 7A).
  • Treatment with 1 ug / ml ZGC-3 did not inhibit cell senescence by adramycin (FIG. 7B).
  • Jangchaechae hexane extract has an effect of inhibiting replication aging due to cell aging and cell division by adriamycin.
  • cytotoxicity of 12 single Jangchaechae single component in human fibroblasts was investigated. Cytotoxicity was not observed in the other materials except ZGC-2 when the compounds were treated with 10 ug / ml (FIG. 11A). Among them, ZGC-5, 6, and 9 were investigated to inhibit cell aging, and it was confirmed that ZGC-9 inhibited cell aging (FIGS. 11B and 11C). When ZGC-9 was treated by concentration, SA- ⁇ -gal activity was reduced in a concentration-dependent manner (Figs. 12A, 12B). It also reduced p53 and pS6 protein expression in a concentration dependent manner (FIG. 12C).
  • ZGC-2 which was toxic when treated at 10 ug / ml, did not exhibit cytotoxicity when treated at 1 ug / ml (FIG. 15A).
  • ZGC-2 was treated at 1 ug / ml to determine how it affects cellular senescence by adriamycin. It was confirmed that cell senescence was inhibited by ZGC-2 (FIGS. 15B and 15C).
  • ZGC-2 inhibited SA- ⁇ -gal activity and p53, p21, pS6 protein expression in a concentration-dependent manner (Figs. 16A, 16B, 16C).
  • ZGC-2 also reduced the amount of intracellular ROS increased by adriamycin at 1 ug / ml (FIG. 17).
  • FIG. 18A and 18B As a result of examining whether ZGC-2 inhibited cell aging even in cells of replication aging, it was observed that the increased SA- ⁇ -gal activity was reduced by the replication aging (FIGS.
  • ZGC-2 ursolic acid
  • ZGC-9 ((-)-bornesitol) isolated from Jang-chae were caused by cell aging and cell division by adriamycin at concentrations of 1 ug / ml and 10 ug / ml, respectively. It was confirmed that there is an effect that inhibits replication aging.

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Abstract

La présente invention concerne une composition pour inhiber la sénescence cellulaire, contenant en tant que principe actif un extrait de Melandrium firmum Rohrbach ou du (-)-bornésitol qui en est séparé, la composition pharmaceutique fournie, destinée à inhiber la sénescence cellulaire, dérivant de l'adriamycine. La présente invention inhibe le processus de sénescence cellulaire des fibroblastes ou des cellules endothéliales de la veine ombilicale, et peut ainsi être utilisée utilement pour traiter des maladies associées à la sénescence, par exemple le vieillissement de la peau, la polyarthrite rhumatoïde, l'ostéoarthrite, l'hépatite, un tissu de dommages chroniques de la peau, l'artériosclérose, l'hyperplasie prostatique et le cancer du foie, etc. On s'attend en outre à ce que la présente invention soit utilisée pour la mise au point d'un aliment fonctionnel antivieillissement, de médicaments contre le vieillissement vasculaire, et de produits cosmétiques qui peuvent inhiber la sénescence cellulaire.
PCT/KR2014/008944 2014-09-25 2014-09-25 Composition pour inhiber la sénescence cellulaire, contenant en tant que principe actif un extrait de melandrium firmum rohrbach ou du bornésitol qui en est séparé Ceased WO2016047823A1 (fr)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004075882A1 (fr) * 2003-02-27 2004-09-10 Ellipsis Biotherapeutics Corporation Procedes permettant de prevenir, traiter et diagnostiquer des troubles dans l'agregation des proteines
KR20060108284A (ko) * 2005-04-12 2006-10-17 (주)젠크로스 장구채 추출물을 포함하는 간 기능 개선 및 간질환 치료용 조성물
KR20110002955A (ko) * 2009-07-03 2011-01-11 김충환 생약추출액이 함유된 샴푸조성물
KR20110097145A (ko) * 2010-02-24 2011-08-31 한국 한의학 연구원 장구채 추출물을 유효성분으로 함유하는 전립선 비대증 예방 및 치료용 조성물
KR20120000246A (ko) * 2010-06-25 2012-01-02 한림대학교 산학협력단 장구채 뿌리 추출물을 포함하는 항암제 조성물
KR20150024605A (ko) * 2013-08-27 2015-03-09 영남대학교 산학협력단 장구채 추출물 또는 이로부터 분리된 보르네시톨을 유효성분으로 함유하는 세포 노화 억제용 조성물

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004075882A1 (fr) * 2003-02-27 2004-09-10 Ellipsis Biotherapeutics Corporation Procedes permettant de prevenir, traiter et diagnostiquer des troubles dans l'agregation des proteines
KR20060108284A (ko) * 2005-04-12 2006-10-17 (주)젠크로스 장구채 추출물을 포함하는 간 기능 개선 및 간질환 치료용 조성물
KR20110002955A (ko) * 2009-07-03 2011-01-11 김충환 생약추출액이 함유된 샴푸조성물
KR20110097145A (ko) * 2010-02-24 2011-08-31 한국 한의학 연구원 장구채 추출물을 유효성분으로 함유하는 전립선 비대증 예방 및 치료용 조성물
KR20120000246A (ko) * 2010-06-25 2012-01-02 한림대학교 산학협력단 장구채 뿌리 추출물을 포함하는 항암제 조성물
KR20150024605A (ko) * 2013-08-27 2015-03-09 영남대학교 산학협력단 장구채 추출물 또는 이로부터 분리된 보르네시톨을 유효성분으로 함유하는 세포 노화 억제용 조성물

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