KR102324233B1 - Novel use of Extract of Solani Nigri Herba - Google Patents
Novel use of Extract of Solani Nigri Herba Download PDFInfo
- Publication number
- KR102324233B1 KR102324233B1 KR1020150034679A KR20150034679A KR102324233B1 KR 102324233 B1 KR102324233 B1 KR 102324233B1 KR 1020150034679 A KR1020150034679 A KR 1020150034679A KR 20150034679 A KR20150034679 A KR 20150034679A KR 102324233 B1 KR102324233 B1 KR 102324233B1
- Authority
- KR
- South Korea
- Prior art keywords
- extract
- yonggyu
- endometriosis
- pharmaceutical composition
- expression
- Prior art date
Links
- 239000000284 extract Substances 0.000 title claims abstract description 159
- 201000009273 Endometriosis Diseases 0.000 claims abstract description 95
- 230000014509 gene expression Effects 0.000 claims description 76
- 239000000469 ethanolic extract Substances 0.000 claims description 48
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 47
- 239000002034 butanolic fraction Substances 0.000 claims description 38
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 36
- 238000011282 treatment Methods 0.000 claims description 27
- 102100026802 72 kDa type IV collagenase Human genes 0.000 claims description 25
- 101710155857 C-C motif chemokine 2 Proteins 0.000 claims description 22
- 102100038280 Prostaglandin G/H synthase 2 Human genes 0.000 claims description 22
- 102100030412 Matrix metalloproteinase-9 Human genes 0.000 claims description 21
- 108050003267 Prostaglandin G/H synthase 2 Proteins 0.000 claims description 21
- 239000000203 mixture Substances 0.000 claims description 20
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims description 19
- 102000004190 Enzymes Human genes 0.000 claims description 18
- 108090000790 Enzymes Proteins 0.000 claims description 18
- 230000005764 inhibitory process Effects 0.000 claims description 18
- 208000002193 Pain Diseases 0.000 claims description 16
- 230000036407 pain Effects 0.000 claims description 16
- 239000008194 pharmaceutical composition Substances 0.000 claims description 16
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 15
- 230000002265 prevention Effects 0.000 claims description 15
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 14
- 102000004127 Cytokines Human genes 0.000 claims description 13
- 108090000695 Cytokines Proteins 0.000 claims description 13
- 239000002904 solvent Substances 0.000 claims description 13
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 12
- 206010061218 Inflammation Diseases 0.000 claims description 12
- 230000004054 inflammatory process Effects 0.000 claims description 12
- 235000013305 food Nutrition 0.000 claims description 11
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 11
- 239000004480 active ingredient Substances 0.000 claims description 10
- 230000002757 inflammatory effect Effects 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 9
- 230000003405 preventing effect Effects 0.000 claims description 9
- 239000000654 additive Substances 0.000 claims description 7
- 230000005012 migration Effects 0.000 claims description 7
- 238000013508 migration Methods 0.000 claims description 7
- 230000000996 additive effect Effects 0.000 claims description 5
- 239000002038 ethyl acetate fraction Substances 0.000 claims description 5
- 238000000605 extraction Methods 0.000 claims description 5
- 239000002044 hexane fraction Substances 0.000 claims description 5
- 239000002037 dichloromethane fraction Substances 0.000 claims description 4
- 208000000509 infertility Diseases 0.000 claims description 4
- 230000036512 infertility Effects 0.000 claims description 4
- 231100000535 infertility Toxicity 0.000 claims description 4
- 208000029082 Pelvic Inflammatory Disease Diseases 0.000 claims description 2
- 206010034238 Pelvic adhesions Diseases 0.000 claims description 2
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 claims description 2
- 206010046798 Uterine leiomyoma Diseases 0.000 claims description 2
- 201000003511 ectopic pregnancy Diseases 0.000 claims description 2
- 201000010260 leiomyoma Diseases 0.000 claims description 2
- 208000025661 ovarian cyst Diseases 0.000 claims description 2
- 230000002018 overexpression Effects 0.000 claims description 2
- 101710151806 72 kDa type IV collagenase Proteins 0.000 claims 2
- 108010015302 Matrix metalloproteinase-9 Proteins 0.000 claims 2
- 102100021943 C-C motif chemokine 2 Human genes 0.000 claims 1
- 210000004027 cell Anatomy 0.000 description 60
- 230000000694 effects Effects 0.000 description 43
- 230000002401 inhibitory effect Effects 0.000 description 43
- 101000627872 Homo sapiens 72 kDa type IV collagenase Proteins 0.000 description 23
- 238000010586 diagram Methods 0.000 description 23
- 102000000018 Chemokine CCL2 Human genes 0.000 description 21
- 238000012790 confirmation Methods 0.000 description 21
- 102100040247 Tumor necrosis factor Human genes 0.000 description 20
- 101000990902 Homo sapiens Matrix metalloproteinase-9 Proteins 0.000 description 19
- 230000021164 cell adhesion Effects 0.000 description 17
- 229940088598 enzyme Drugs 0.000 description 17
- 238000002474 experimental method Methods 0.000 description 16
- 238000002360 preparation method Methods 0.000 description 16
- 230000012292 cell migration Effects 0.000 description 14
- 108020004999 messenger RNA Proteins 0.000 description 14
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 13
- 238000003753 real-time PCR Methods 0.000 description 12
- 238000002513 implantation Methods 0.000 description 11
- 230000006872 improvement Effects 0.000 description 9
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 8
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 8
- 230000000692 anti-sense effect Effects 0.000 description 6
- 230000036541 health Effects 0.000 description 6
- 230000006698 induction Effects 0.000 description 6
- 210000004303 peritoneum Anatomy 0.000 description 6
- 238000003757 reverse transcription PCR Methods 0.000 description 6
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 238000009472 formulation Methods 0.000 description 4
- 235000013376 functional food Nutrition 0.000 description 4
- 230000035515 penetration Effects 0.000 description 4
- 239000000546 pharmaceutical excipient Substances 0.000 description 4
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 4
- 230000001629 suppression Effects 0.000 description 4
- 230000009772 tissue formation Effects 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 244000061457 Solanum nigrum Species 0.000 description 3
- 235000002594 Solanum nigrum Nutrition 0.000 description 3
- 210000000683 abdominal cavity Anatomy 0.000 description 3
- 239000002775 capsule Substances 0.000 description 3
- 239000002299 complementary DNA Substances 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
- 235000008504 concentrate Nutrition 0.000 description 3
- 238000012937 correction Methods 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 239000000839 emulsion Substances 0.000 description 3
- 238000007912 intraperitoneal administration Methods 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000002560 therapeutic procedure Methods 0.000 description 3
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- NIPNSKYNPDTRPC-UHFFFAOYSA-N N-[2-oxo-2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 NIPNSKYNPDTRPC-UHFFFAOYSA-N 0.000 description 2
- AFCARXCZXQIEQB-UHFFFAOYSA-N N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CCNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 AFCARXCZXQIEQB-UHFFFAOYSA-N 0.000 description 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
- RJKFOVLPORLFTN-LEKSSAKUSA-N Progesterone Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 RJKFOVLPORLFTN-LEKSSAKUSA-N 0.000 description 2
- 210000001015 abdomen Anatomy 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 239000003125 aqueous solvent Substances 0.000 description 2
- 239000007900 aqueous suspension Substances 0.000 description 2
- 235000013361 beverage Nutrition 0.000 description 2
- AZFLJNIPTRTECV-FUMNGEBKSA-N dienogest Chemical compound C1CC(=O)C=C2CC[C@@H]([C@H]3[C@@](C)([C@](CC3)(O)CC#N)CC3)C3=C21 AZFLJNIPTRTECV-FUMNGEBKSA-N 0.000 description 2
- 229960003309 dienogest Drugs 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 235000013373 food additive Nutrition 0.000 description 2
- 239000002778 food additive Substances 0.000 description 2
- 238000005194 fractionation Methods 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 description 2
- 239000007928 intraperitoneal injection Substances 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 239000011777 magnesium Substances 0.000 description 2
- 229910052749 magnesium Inorganic materials 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000027758 ovulation cycle Effects 0.000 description 2
- 229940056360 penicillin g Drugs 0.000 description 2
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 2
- 229920000053 polysorbate 80 Polymers 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000829 suppository Substances 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 210000004291 uterus Anatomy 0.000 description 2
- 239000003981 vehicle Substances 0.000 description 2
- 239000000080 wetting agent Substances 0.000 description 2
- WBYWAXJHAXSJNI-VOTSOKGWSA-M .beta-Phenylacrylic acid Natural products [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 description 1
- KZDCMKVLEYCGQX-UDPGNSCCSA-N 2-(diethylamino)ethyl 4-aminobenzoate;(2s,5r,6r)-3,3-dimethyl-7-oxo-6-[(2-phenylacetyl)amino]-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid;hydrate Chemical compound O.CCN(CC)CCOC(=O)C1=CC=C(N)C=C1.N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 KZDCMKVLEYCGQX-UDPGNSCCSA-N 0.000 description 1
- YLZOPXRUQYQQID-UHFFFAOYSA-N 3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)-1-[4-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]piperazin-1-yl]propan-1-one Chemical compound N1N=NC=2CN(CCC=21)CCC(=O)N1CCN(CC1)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F YLZOPXRUQYQQID-UHFFFAOYSA-N 0.000 description 1
- IPJDHSYCSQAODE-UHFFFAOYSA-N 5-chloromethylfluorescein diacetate Chemical compound O1C(=O)C2=CC(CCl)=CC=C2C21C1=CC=C(OC(C)=O)C=C1OC1=CC(OC(=O)C)=CC=C21 IPJDHSYCSQAODE-UHFFFAOYSA-N 0.000 description 1
- 208000030507 AIDS Diseases 0.000 description 1
- 208000002874 Acne Vulgaris Diseases 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 208000004746 Atrophic Vaginitis Diseases 0.000 description 1
- 206010003693 Atrophic vulvovaginitis Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- WBYWAXJHAXSJNI-SREVYHEPSA-N Cinnamic acid Chemical compound OC(=O)\C=C/C1=CC=CC=C1 WBYWAXJHAXSJNI-SREVYHEPSA-N 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 108010037462 Cyclooxygenase 2 Proteins 0.000 description 1
- 206010013935 Dysmenorrhoea Diseases 0.000 description 1
- 208000004483 Dyspareunia Diseases 0.000 description 1
- LVGKNOAMLMIIKO-UHFFFAOYSA-N Elaidinsaeure-aethylester Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC LVGKNOAMLMIIKO-UHFFFAOYSA-N 0.000 description 1
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 1
- 239000004606 Fillers/Extenders Substances 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- NMJREATYWWNIKX-UHFFFAOYSA-N GnRH Chemical compound C1CCC(C(=O)NCC(N)=O)N1C(=O)C(CC(C)C)NC(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)CNC(=O)C(NC(=O)C(CO)NC(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)C(CC=1NC=NC=1)NC(=O)C1NC(=O)CC1)CC1=CC=C(O)C=C1 NMJREATYWWNIKX-UHFFFAOYSA-N 0.000 description 1
- 229920002907 Guar gum Polymers 0.000 description 1
- 206010019027 Haemothorax Diseases 0.000 description 1
- 206010020112 Hirsutism Diseases 0.000 description 1
- 208000033830 Hot Flashes Diseases 0.000 description 1
- 206010060800 Hot flush Diseases 0.000 description 1
- 208000007101 Muscle Cramp Diseases 0.000 description 1
- MKYBYDHXWVHEJW-UHFFFAOYSA-N N-[1-oxo-1-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propan-2-yl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(C(C)NC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 MKYBYDHXWVHEJW-UHFFFAOYSA-N 0.000 description 1
- 229930182559 Natural dye Natural products 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 206010039792 Seborrhoea Diseases 0.000 description 1
- 241000208292 Solanaceae Species 0.000 description 1
- 208000005392 Spasm Diseases 0.000 description 1
- 101000857870 Squalus acanthias Gonadoliberin Proteins 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 244000299461 Theobroma cacao Species 0.000 description 1
- 235000005764 Theobroma cacao ssp. cacao Nutrition 0.000 description 1
- 235000005767 Theobroma cacao ssp. sphaerocarpum Nutrition 0.000 description 1
- 206010046406 Ureteric obstruction Diseases 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 206010000496 acne Diseases 0.000 description 1
- 230000000202 analgesic effect Effects 0.000 description 1
- 229940035676 analgesics Drugs 0.000 description 1
- 239000000730 antalgic agent Substances 0.000 description 1
- 229940124599 anti-inflammatory drug Drugs 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000008366 buffered solution Substances 0.000 description 1
- 235000014121 butter Nutrition 0.000 description 1
- 235000001046 cacaotero Nutrition 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 229930016911 cinnamic acid Natural products 0.000 description 1
- 235000013985 cinnamic acid Nutrition 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000008094 contradictory effect Effects 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- POZRVZJJTULAOH-LHZXLZLDSA-N danazol Chemical compound C1[C@]2(C)[C@H]3CC[C@](C)([C@](CC4)(O)C#C)[C@@H]4[C@@H]3CCC2=CC2=C1C=NO2 POZRVZJJTULAOH-LHZXLZLDSA-N 0.000 description 1
- -1 depigmentation Chemical compound 0.000 description 1
- 230000035614 depigmentation Effects 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 230000002996 emotional effect Effects 0.000 description 1
- 210000005168 endometrial cell Anatomy 0.000 description 1
- 230000002357 endometrial effect Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- LVGKNOAMLMIIKO-QXMHVHEDSA-N ethyl oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC LVGKNOAMLMIIKO-QXMHVHEDSA-N 0.000 description 1
- 229940093471 ethyl oleate Drugs 0.000 description 1
- 206010016256 fatigue Diseases 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 239000007903 gelatin capsule Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000000665 guar gum Substances 0.000 description 1
- 229960002154 guar gum Drugs 0.000 description 1
- 235000010417 guar gum Nutrition 0.000 description 1
- 208000006750 hematuria Diseases 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 238000001794 hormone therapy Methods 0.000 description 1
- 229960000890 hydrocortisone Drugs 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 238000010874 in vitro model Methods 0.000 description 1
- 208000021267 infertility disease Diseases 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- VMPHSYLJUKZBJJ-UHFFFAOYSA-N lauric acid triglyceride Natural products CCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCC)COC(=O)CCCCCCCCCCC VMPHSYLJUKZBJJ-UHFFFAOYSA-N 0.000 description 1
- 229940069445 licorice extract Drugs 0.000 description 1
- 239000012669 liquid formulation Substances 0.000 description 1
- 229940057995 liquid paraffin Drugs 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 229960003511 macrogol Drugs 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- WBYWAXJHAXSJNI-UHFFFAOYSA-N methyl p-hydroxycinnamate Natural products OC(=O)C=CC1=CC=CC=C1 WBYWAXJHAXSJNI-UHFFFAOYSA-N 0.000 description 1
- 238000010232 migration assay Methods 0.000 description 1
- 239000000978 natural dye Substances 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000037312 oily skin Effects 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 210000003245 peritoneal mesothelial cell Anatomy 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 201000003144 pneumothorax Diseases 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 201000010808 postmenopausal atrophic vaginitis Diseases 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000186 progesterone Substances 0.000 description 1
- 229960003387 progesterone Drugs 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 235000011083 sodium citrates Nutrition 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 210000003932 urinary bladder Anatomy 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 230000004584 weight gain Effects 0.000 description 1
- 235000019786 weight gain Nutrition 0.000 description 1
Images
Classifications
-
- 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)
- A61K36/81—Solanaceae (Potato family), e.g. tobacco, nightshade, tomato, belladonna, capsicum or jimsonweed
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/02—Drugs for genital or sexual disorders; Contraceptives for disorders of the vagina
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Botany (AREA)
- Mycology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Biotechnology (AREA)
- Alternative & Traditional Medicine (AREA)
- Epidemiology (AREA)
- Microbiology (AREA)
- Medical Informatics (AREA)
- Nutrition Science (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Gynecology & Obstetrics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Reproductive Health (AREA)
- Endocrinology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicines Containing Plant Substances (AREA)
- Steroid Compounds (AREA)
Abstract
본 발명은 용규 추출물의 신규 용도를 제공한다. 본 발명에 의해 자궁내막증 또는 이의 합병증을 효과적으로 치료, 개선 및/또는 예방할 수 있다는 장점이 있다.The present invention provides a novel use of the Yonggyu extract. The present invention has the advantage that endometriosis or its complications can be effectively treated, improved and/or prevented.
Description
본 발명은 용규 추출물에 관한 것으로, 보다 상세하게는 용규 추출물의 신규 용도에 관한 것이다.The present invention relates to a Yonggyu extract, and more particularly, to a novel use of the Yonggyu extract.
자궁내막증(endometriosis)은 가임기 여성의 3~10%, 불임 여성의 25~35% 정도에서 나타나는 매우 흔한 부인과 질환으로 자궁내막조직이 자궁 이외에 장소(난소, 복강, 장관, 방광)에서 비정상적으로 증식하는 질환이다.Endometriosis is a very common gynecological disease that occurs in 3-10% of women of childbearing age and 25-35% of infertile women. is a disease
자궁내막증은 월경통(dysmenorrhoea), 성교통(dyspareunia) 및 침범부위에 따른 다양한 통증 (ex. 폐침범시: 기흉, 혈흉/ 요관침범시: 요관폐쇄, 혈뇨)을 증상으로 하며, 30-50%의 환자에서 불임의 원인으로 확인되고 있다.Endometriosis is characterized by dysmenorrhoea, dyspareunia, and various pains depending on the site of involvement (ex. pulmonary involvement: pneumothorax, hemothorax/ureteral involvement: ureter obstruction, hematuria), accounting for 30-50% of patients. has been identified as a cause of infertility.
현재까지 수술을 통한 자궁내막 제거수술 외에 자궁내막증에 대한 근본적인 치료법이 없으며, 이러한 수술요법은 차후 임신에 제약을 줄 수 있으며 재발율 또한 높은 편이다.To date, there is no fundamental treatment for endometriosis other than surgical removal of endometriosis.
현재 내과적 치료로는 통증완화를 위해 진통제, 소염제를 사용하는 보존적 요법과 다나졸, 프로게스테론, GnRH를 사용하여 생리주기를 제어하는 호르몬치료가 사용되고 있으나 이들은 근본적 치료법이 아닌 증상의 개선일 뿐이다.Currently, as a medical treatment, conservative therapy using analgesics and anti-inflammatory drugs for pain relief, and hormone therapy using danazole, progesterone, and GnRH to control the menstrual cycle are used, but these are only symptomatic improvements, not fundamental treatments.
또한 이들 호르몬의 장기적 사용은 여러 가지 부작용 (체중의 증가, 수분 축적, 피로, 여드름, 지성 피부, 조모증, 위축성 질염, 안면 홍조, 근육 경련, 불안정한 감정 상태, 간세포 독성)을 야기하며, 게다가 이러한 치료에 대한 재발률도 매우 높은 것으로 알려져 있다.In addition, long-term use of these hormones causes a number of side effects (weight gain, water accumulation, fatigue, acne, oily skin, hirsutism, atrophic vaginitis, hot flashes, muscle spasms, unstable emotional states, hepatocytotoxicity). It is also known that the recurrence rate for treatment is very high.
이처럼 자궁내막증은 치명적이지는 않지만 극심한 통증으로 인해 일상생활에 불편함이 있고 심한 경우 불임에까지 이르는 질환으로, 여성의 삶의 질과 임신의 문제에서 심각하게 인식되어야 하고 반드시 극복해야 할 질병임에도 불구하고 아직까지 이렇다 할 예방 및 치료방법이 존재하지 않고 있다.As such, endometriosis is not fatal, but it is a disease that causes discomfort in daily life due to extreme pain and even leads to infertility in severe cases. So far, there is no such prevention and treatment method.
따라서 여성, 특히 가임기 여성들에게 침습적인(invasive) 수술요법이나 단순한 진통요법, 인위적인 생리주기제어법 등이 아닌 자연적이면서도 부작용이 낮은 새로운 개념의 자궁내막증 예방, 치료제의 개발이 시급한 실정이다.Therefore, it is urgent for women, especially women of childbearing age, to develop a new concept of prevention and treatment for endometriosis, which is natural and has low side effects, rather than invasive surgical therapy, simple analgesic therapy, or artificial menstrual cycle control.
한편, 용규 추출물은 AIDS 치료 효과를 가짐이 알려져 있다(대한민국 공개특허 제1020060034178호). On the other hand, Yonggyu extract is known to have an AIDS treatment effect (Korean Patent Publication No. 1020060034178).
본 발명이 해결하고자 하는 과제는 용규 추출물의 신규 용도를 제공하는 것이다.The problem to be solved by the present invention is to provide a novel use of the Yonggyu extract.
본 발명의 과제는 상기에 언급된 과제로 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당업자에게 명확하게 이해될 수 있을 것이다.The problems of the present invention are not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.
본 발명자들은 놀랍게도 용규 추출물이 자궁내막증 또는 이의 합병증 치료, 개선 및/또는 예방에 효과를 나타냄을 알게 되어 본 발명을 완성하였다.The present inventors have surprisingly found that Yonggyu extract is effective in treating, improving and/or preventing endometriosis or its complications, and thus completed the present invention.
본 발명은 용규 추출물의 자궁내막증 또는 이의 합병증 치료, 개선 및/또는 예방을 위한 의약 및/또는 식품 용도를 제공한다.The present invention provides a pharmaceutical and/or food use of the Yonggyu extract for the treatment, improvement and/or prevention of endometriosis or its complications.
또한, 본 발명은 용규 추출물을 유효성분으로 포함하는 것을 특징으로 하는 자궁내막증 또는 이의 합병증 치료 또는 예방용 약학 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for the treatment or prevention of endometriosis or its complications, characterized in that it contains Yonggyu extract as an active ingredient.
상기 용규(Solani Nigri Herba)는 대한민국약전외한약(생약)규격집에 수재된 것으로, 솔라눔니그럼{Solanum nigrum Linne(가지과 Solanaceae)}의 지상부일 수 있다. 지상부는 줄기와 잎을 포함하는 의미로, 꽃도 포함될 수 있다.The Yong-gyu (Solani Nigri Herba) is listed in the Korean Non-Pharmaceutical Herbal Medicine (Germany) Standards, and may be an above-ground part of Solanum nigrum Linne (Solanaceae). The above-ground part is meant to include stems and leaves, and flowers may also be included.
상기 약학조성물은 약제학적으로 허용되는 첨가제를 더 포함하고, 상기 용규 추출물 및 상기 첨가제로 이루어질 수 있다.The pharmaceutical composition may further include a pharmaceutically acceptable additive, and may consist of the Yonggyu extract and the additive.
상기 자궁내막증 또는 이의 합병증은 MMP-2, MMP-9, COX-2, TNF-알파(α), 또는 MCP-1 중에서 선택된 하나 이상의 과발현에서 기인하는 것일 수 있다.The endometriosis or its complications may be due to overexpression of one or more selected from MMP-2, MMP-9, COX-2, TNF-alpha (α), or MCP-1.
상기 합병증은 골반염, 골반유착, 난소낭종, 자궁근종, 자궁외임신 및 불임으로 이루어진 군에서 선택된 하나 이상일 수 있다.The complication may be one or more selected from the group consisting of pelvic inflammation, pelvic adhesions, ovarian cyst, uterine fibroids, ectopic pregnancy, and infertility.
상기 용규 추출물은 상기 용규를 용매로 추출한 용매추출물로, 상기 용매는 물, 알코올, 또는 그 혼합물 중에서 선택된 하나 이상일 수 있다.The Yonggyu extract is a solvent extract obtained by extracting the Yonggyu with a solvent, and the solvent may be one or more selected from water, alcohol, or a mixture thereof.
상기 알코올은 탄소수 1~5의 저급알코올일 수 있다.The alcohol may be a lower alcohol having 1 to 5 carbon atoms.
상기 용매는 70부피% 에탄올일 수 있다.The solvent may be 70% by volume ethanol.
상기 용규 추출물은 상기 용규를 70부피%에탄올로 추출한 70부피%에탄올추출물을 부탄올로 분획추출한 부탄올분획추출물일 수 있다.The Yonggyu extract may be a butanol fractional extract obtained by fractionally extracting the 70% by volume ethanol extract obtained by extracting the Yonggyu with 70% by volume ethanol with butanol.
상기 부탄올분획추출물은 (A) 용규를 70부피%에탄올로 추출한 70부피%에탄올추출물을 준비하는 단계; (B) 상기 (A)단계의 70부피%에탄올추출물에 물을 가하는 단계; (C) 상기 (B)단계의 물이 가해진 70부피%에탄올추출물을 n-헥산으로 분획추출하는 단계; (D) 상기 (C)단계의 n-헥산 분획을 제거하고 남은 수층을 디클로로메탄으로 분획추출하는 단계; (E) 상기 (D)단계의 디클로로메탄 분획을 제거하고 남은 수층을 에틸아세테이트로 분획추출하는 단계; 및 (F) 상기 (E)단계의 에틸아세테이트 분획을 제거하고 남은 수층을 부탄올로 분획추출하는 단계에 의해 수득한 분획추출물일 수 있다.The butanol fraction extract is prepared by (A) preparing a 70 vol % ethanol extract obtained by extracting Yonggyu with 70 vol % ethanol; (B) adding water to the 70% by volume ethanol extract of step (A); (C) fractional extraction of the 70% by volume ethanol extract to which water was added in step (B) with n-hexane; (D) removing the n-hexane fraction of step (C) and fraction-extracting the remaining aqueous layer with dichloromethane; (E) removing the dichloromethane fraction of step (D) and fractionally extracting the remaining aqueous layer with ethyl acetate; and (F) removing the ethyl acetate fraction of step (E) and fractionally extracting the remaining aqueous layer with butanol.
상기 치료 또는 예방은 자궁내막증 세포의 부착 또는 이동 억제에 의한 것일 수 있다.The treatment or prevention may be by inhibition of adhesion or migration of endometriosis cells.
상기 부착 또는 이동 억제는 MMP-2 또는 MMP-9 중에서 선택된 하나 이상의 발현 억제에 의한 것일 수 있다.The inhibition of adhesion or migration may be due to inhibition of expression of one or more selected from MMP-2 or MMP-9.
상기 치료 또는 예방은 염증 및 통증 관련 효소 발현 억제 또는 염증성 싸이토카인 발현 억제 중에서 선택된 하나 이상에 의한 것일 수 있다.The treatment or prevention may be by one or more selected from inhibition of expression of inflammation and pain-related enzymes or inhibition of inflammatory cytokine expression.
또한, 본 발명은 용규 추출물을 유효성분으로 포함하는 것을 특징으로 하는 자궁내막증 또는 이의 합병증 개선 또는 예방용 식품 조성물을 제공한다.In addition, the present invention provides a food composition for improving or preventing endometriosis or its complications, characterized in that it contains Yonggyu extract as an active ingredient.
별도의 언급이 없는 한 상기 식품 조성물은 본 발명의 약학조성물에서 언급된 사항이 모순되지 않는 한 동일하게 적용된다. 상기 '개선'은 '치료'에 포함되며, 상태 또는 증세가 호전되는 것을 의미한다.Unless otherwise stated, the food composition is equally applied as long as the matters mentioned in the pharmaceutical composition of the present invention do not contradict each other. The 'improvement' is included in 'treatment' and means that a condition or symptom is improved.
상기 식품조성물은 음료를 포함하는 식품에 다양하게 포함될 수 있고, 음료, 껌, 차, 건강기능식품 등의 형태일 수 있으며, 상기 건강기능식품은 정제, 캡슐제 등의 제형으로 제제화할 수 있다. 상기 건강기능식품이라 함은 대한민국 건강기능식품에 관한 법률 제12669호에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조(가공을 포함한다. 이하 같다)한 식품을 말하며, "기능성"이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻는 것을 말한다. 상기 식품 조성물은 통상의 식품 첨가물을 포함할 수 있으며, 상기 식품 첨가물은 케톤류, 글리신, 구연산나트륨, 니코틴산, 계피산 등의 화학적 합성물, 감색소, 감초추출물, 결정셀룰로오스, 고량색소, 구아검 등의 천연첨가물, L-글루타민산나트륨 제제, 면류첨가알칼리제, 보존료제제, 타르색소제제 등의 혼합제제류들을 들 수 있다.The food composition may be variously included in food including beverages, and may be in the form of beverages, gum, tea, health functional food, etc., and the health functional food may be formulated in dosage forms such as tablets and capsules. The term "health functional food" refers to a food manufactured (including processing; hereinafter the same) using raw materials or ingredients useful for the human body in accordance with Act No. 12669 on Health Functional Foods of the Republic of Korea, and "functionality" It refers to obtaining useful effects for health purposes such as regulating nutrients or physiological effects on the structure and function of the human body. The food composition may include conventional food additives, and the food additives are chemical compounds such as ketones, glycine, sodium citrate, nicotinic acid, and cinnamic acid, natural dyes such as depigmentation, licorice extract, crystalline cellulose, high pigment, and guar gum. Mixed preparations such as additives, sodium L-glutamate preparations, noodles added alkalis, preservatives, and tar dye preparations may be mentioned.
본 발명은 또한 용규 추출물을 투여를 필요로 하는 인간을 포함한 포유류에게 용규 추출물을 투여하는 단계를 포함하는 자궁내막증 또는 이의 합병증의 치료 또는 예방법을 제공하며, 용규 추출물의 자궁내막증 또는 이의 합병증의 치료 또는 예방용 제제 제조를 위한 용도를 제공한다. 상기 투여되는 용규 추출물은 유효량의 용규 추출물일 수 있다.The present invention also provides a method for treating or preventing endometriosis or its complications, comprising administering the Yonggyu extract to mammals, including humans, in need of administration of the Yonggyu extract. Provided is a use for the manufacture of a prophylactic formulation. The administered Yonggyu extract may be an effective amount of Yonggyu extract.
별도의 언급이 없는 한, 본 발명의 약학 조성물에서 언급된 사항은 모순되지 않는 한 본 발명의 방법, 및 용도에도 동일하게 적용된다. Unless otherwise stated, the matters mentioned in the pharmaceutical composition of the present invention apply equally to the method and use of the present invention unless contradictory.
상기 용규 추출물 또는 조성물은 인간을 포함한 포유류에 경구 또는 비경구로 투여가 가능하며, 유효성분을 약학적으로 허용되는 첨가제와 함께 배합하여 제제화하여 투여할 수 있다. 제제화할 경우 보통 사용하는 첨가제는 유효성분 이외의 물질로 담체, 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제, 희석제, 부형제 등일 수 있다. 경구투여를 위한 고형제제는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 본 발명의 조성물에 적어도 하나 이상의 부형제 예를 들면, 전분, 탄산칼슘, 수크로스, 락토오스, 및/또는 젤라틴 등을 첨가하여 제조한다. 또한, 마그네슘, 탈크 등 윤활제도 사용된다. 경구투여를 위한 액상제제로는 현탁제, 내용액제, 유제 및 시럽제 등이 해당되며, 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러가지 부형제, 예를 들면 습윤제, 감미제, 방향제 및/또는 보존제 등이 포함될 수 있다. 비경구투여를 위한 제제에는 주사가능한 액제, 현탁제, 유제, 동결건조제, 비강세척제 및 좌제가 포함된다. 주사가능한 액제, 현탁제, 유제는 물, 비수성용제나 현탁용제와 유효성분을 혼합하여 제조할 수 있으며, 비수성용제와 현탁용제로는 프로필렌글리콜, 폴리에틸렌글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사가능한 에스테르 등일 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈 61, 카카오지, 라우린지, 글리세롤 및/또는 젤라틴 등이 사용될 수 있다. 비경구 투여시 피하주사, 정맥주사 또는 근육내 주사로 투여가능하다.The Yonggyu extract or composition can be administered orally or parenterally to mammals including humans, and the active ingredient can be formulated and administered with a pharmaceutically acceptable additive. In the case of formulation, additives usually used are substances other than active ingredients, and may be carriers, fillers, extenders, binders, wetting agents, disintegrants, surfactants, diluents, excipients, and the like. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations include at least one excipient in the composition of the present invention, for example, starch, calcium carbonate, sucrose, lactose, and / or by adding gelatin or the like. Lubricating agents such as magnesium and talc are also used. Liquid formulations for oral administration include suspensions, internal solutions, emulsions and syrups, and various excipients, such as wetting agents, sweeteners, fragrances and/or preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. may be included. Formulations for parenteral administration include injectable solutions, suspensions, emulsions, lyophilisates, nasal washes and suppositories. Injectable solutions, suspensions, and emulsions can be prepared by mixing water, non-aqueous solvents or suspensions with active ingredients, and non-aqueous solvents and suspensions include vegetable oils such as propylene glycol, polyethylene glycol, olive oil, and ethyl oleate. and injectable esters such as As the base of the suppository, witepsol, macrogol, Tween 61, cacao butter, laurin fat, glycerol and/or gelatin, etc. may be used. When administered parenterally, it can be administered by subcutaneous injection, intravenous injection or intramuscular injection.
본 발명의 조성물에 포함되거나, 용도 및 방법 중 사용되는 용규 추출물은 성인 여성 기준으로 1일 0.0001 ~ 1000mg/kg, 바람직하게는 0.0001 ~ 100 mg/kg, 보다 바람직하게는 0.001~10mg/kg의 용량으로 사용가능하다. 투여는 하루에 한번 또는 수회로 나누어 투여할 수 있다. 그러나, 본 발명의 범주는 상기 투여량 및 투여횟수에 의해 제한되지 않는다.Yonggyu extract included in the composition of the present invention or used in uses and methods is 0.0001 to 1000 mg/kg, preferably 0.0001 to 100 mg/kg, more preferably 0.001 to 10 mg/kg per day based on adult females. can be used as Administration may be administered once or divided into several times a day. However, the scope of the present invention is not limited by the dosage and number of administration.
본 발명의 조성물은 상기 유효성분을 조성물 총 중량에 대하여 0.1~99.9중량% 함유할 수 있다.The composition of the present invention may contain 0.1 to 99.9% by weight of the active ingredient based on the total weight of the composition.
상기 용규 추출물은 약학적으로 또는 식품학적으로 허용되는 담체, 부형제 또는 희석제 등을 첨가하여 제제화할 수 있으며, 제제화에 관한 내용은 Remington's Pharmaceutical Science(최근판), Mack Publishing Company, Easton PA 등의 문헌을 참조할 수 있다.The Yonggyu extract can be formulated by adding a pharmaceutically or pharmaceutically acceptable carrier, excipient or diluent, etc. For information on formulation, see Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA, etc. can refer to
본 발명에 의해 자궁내막증 또는 이의 합병증을 효과적으로 치료, 개선 및/또는 예방할 수 있다.The present invention can effectively treat, improve and/or prevent endometriosis or its complications.
도 1은 용규 알콜 추출물의 MMP-2 발현 억제 효과 확인 실험 결과를 나타낸 도이다.
도 2는 용규 알콜 추출물의 MMP-9 발현 억제 효과 확인 실험 결과를 나타낸 도이다.
도 3은 용규 알콜 추출물의 COX-2 발현 억제 효과 확인 실험 결과를 나타낸 도이다.
도 4는 용규 알콜 추출물의 TNF알파 발현 억제 효과 확인 실험 결과를 나타낸 도이다.
도 5는 용규 알콜 추출물의 MCP-1 발현 억제 효과 확인 실험 결과를 나타낸 도이다.
도 6은 용규 알콜 추출물의 자궁내막증 세포 부착 및 착상 억제 효과 확인 실험 결과를 나타낸 도이다.
도 7은 용규 알콜 추출물의 자궁내막증 세포 이동 억제 효과 확인 실험 결과를 나타낸 도이다.
도 8은 용규 알콜 추출물의 복강내투여모델에서 효과 확인 실험 결과를 나타낸 도이다.
도 9는 용규 알콜 추출물의 수술유도모델에서 효과 확인 실험 결과를 나타낸 도이다.
도 10은 용규 부탄올분획 추출물의 MMP-2 발현 억제 효과 확인 실험 결과를 나타낸 도이다.
도 11은 용규 부탄올분획 추출물의 MMP-9 발현 억제 효과 확인 실험 결과를 나타낸 도이다.
도 12는 용규 부탄올분획 추출물의 COX-2 발현 억제 효과 확인 실험 결과를 나타낸 도이다.
도 13은 용규 부탄올분획 추출물의 TNF알파 발현 억제 효과 확인 실험 결과를 나타낸 도이다.
도 14는 용규 부탄올분획 추출물의 MCP-1 발현 억제 효과 확인 실험 결과를 나타낸 도이다.
도 15는 용규 부탄올분획 추출물의 자궁내막증 세포 부착 및 착상 억제 효과 확인 실험 결과를 나타낸 도이다.
도 16은 용규 물 추출물의 MMP-2 발현 억제 효과 확인 실험 결과를 나타낸 도이다.
도 17은 용규 물 추출물의 COX-2 발현 억제 효과 확인 실험 결과를 나타낸 도이다.
도 18은 용규 물 추출물의 TNF알파 발현 억제 효과 확인 실험 결과를 나타낸 도이다.
도 19는 용규 물 추출물의 MCP-1 발현 억제 효과 확인 실험 결과를 나타낸 도이다.1 is a diagram showing the results of experiments confirming the MMP-2 expression inhibitory effect of Yonggyu alcohol extract.
2 is a diagram showing the results of experiments confirming the MMP-9 expression inhibitory effect of Yonggyu alcohol extract.
3 is a diagram showing the results of experiments confirming the effect of inhibiting COX-2 expression of Yonggyu alcohol extract.
Figure 4 is a diagram showing the experimental results confirming the TNF alpha expression inhibitory effect of Yonggyu alcohol extract.
5 is a diagram showing the results of experiments confirming the MCP-1 expression inhibitory effect of Yonggyu alcohol extract.
6 is a diagram showing the results of experiments confirming the effect of Yonggyu alcohol extract to inhibit endometriosis cell adhesion and implantation.
7 is a diagram showing the results of experiments confirming the effect of Yonggyu alcohol extract on the inhibition of endometriosis cell migration.
8 is a diagram showing the experimental results of confirming the effect in the intraperitoneal administration model of Yonggyu alcohol extract.
9 is a diagram showing the experimental results of confirming the effect in the surgical induction model of Yonggyu alcohol extract.
10 is a diagram showing the results of experiments confirming the MMP-2 expression inhibitory effect of the Yonggyu butanol fraction extract.
11 is a diagram showing the results of experiments confirming the MMP-9 expression inhibitory effect of Yonggyu butanol fraction extract.
12 is a view showing the results of experiments confirming the effect of inhibiting the expression of COX-2 of the extract of Yonggyu butanol fraction.
13 is a diagram showing the results of experiments confirming the TNF alpha expression inhibitory effect of Yonggyu butanol fraction extract.
14 is a diagram showing the results of experiments confirming the MCP-1 expression inhibitory effect of Yonggyu butanol fraction extract.
15 is a diagram showing the results of experiments confirming the effect of the extract of Yonggyu butanol fraction on endometriosis cell adhesion and implantation inhibition.
16 is a diagram showing the results of experiments confirming the MMP-2 expression inhibitory effect of the Yonggyu water extract.
17 is a diagram showing the results of the experiment confirming the effect of inhibiting the expression of COX-2 of the water extract of Yonggyu.
18 is a diagram showing the results of experiments confirming the TNF alpha expression inhibitory effect of Yonggyu water extract.
19 is a diagram showing the results of experiments confirming the MCP-1 expression inhibitory effect of the Yonggyu water extract.
이하, 실시예, 제조예 및 도면을 참조하여, 본 발명을 보다 상세하게 설명하나, 하기 실시예 및 제조예는 본 발명을 예시하기 위한 것일 뿐으로 본 발명의 내용이 하기 실시예, 제조예 및 도면에 의해 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to Examples, Preparation Examples and Drawings, but the following Examples and Preparation Examples are only for illustrating the present invention and the content of the present invention is the following Examples, Preparation Examples and Drawings is not limited by
자궁내막증의 특징인 세포이동 및 부착 관련 효소{MMP-2(matrix metallopeptidase-2) 및/또는 MMP-9(matrix metallopeptidase-9)}의 발현 변화, 염증 및 통증 관련 효소{COX-2(cyclooxygenase-2)}의 발현 변화, 염증성 싸이토카인{TNF-알파(tumor necrosis factor alpha) 및/또는 MCP-1(Monocyte chemotactic protein-1)}의 발현 변화, 자궁내막증세포 부착 및 착상 변화, 자궁내막증세포 이동 변화 등에 용규 추출물이 미치는 영향을 확인함으로써, 용규 추출물이 자궁내막증 또는 이의 합병증 치료, 개선 또는 예방 효과를 가짐을 아래와 같은 방법으로 확인하였다.
Cell migration and adhesion-related enzymes characteristic of endometriosis {MMP-2 (matrix metallopeptidase-2) and/or MMP-9 (matrix metallopeptidase-9)} expression changes, inflammation and pain-related enzymes {COX-2 (cyclooxygenase- 2)} expression change, inflammatory cytokine {TNF-alpha (tumor necrosis factor alpha) and/or MCP-1 (Monocyte chemotactic protein-1)} expression change, endometriosis cell adhesion and implantation change, endometriosis cell migration change By confirming the effect of the Yonggyu extract on the back, it was confirmed that the Yonggyu extract has an effect of treating, improving or preventing endometriosis or its complications by the following method.
하기 실시예 중 기재된 자궁내막증 모델 세포는 자궁내막증 여성의 활성 자궁내막증 조직을 이용하여 개발된, human immortalized endometriotic epithelial cells인 12Z 세포주를 준비하였다{Johann-Wolfgang-Goethe-Universitaet(Germany)의 Dr. Starzinski-Powitz 제공}. 이 세포는 기존에 알려진 활성 자궁내막증 조직(active endometotric tissue)의 분자생물학적 성격과 유사함이 증명되어(Banu et al., 2008), 자궁내막증의 분자생물학적 연구에 대한 새로운 인 비트로 모델로 사용되고 있다. 세포는 RPMI 1640 및 DMEM/F12 배지(10% 우태아혈청, 100 U/ml 페니실린 G, 100g/ml 스트렙토마이신 첨가됨; 라이프 테크놀로지, 그랜드 아일랜드, 뉴욕) 중, 섭씨 37도, 5%이산화탄소-95%공기 조건에서 유지되었다.
The endometriosis model cells described in the Examples below were prepared using a 12Z cell line, which is human immortalized endometriotic epithelial cells, developed using active endometriosis tissue from women with endometriosis {Dr. Johann-Wolfgang-Goethe-Universitaet (Germany). Courtesy of Starzinski-Powitz}. These cells have been proven to be similar to the molecular biological properties of previously known active endometotric tissues (Banu et al., 2008), and are being used as a new in vitro model for molecular biological studies of endometriosis. Cells were cultured in RPMI 1640 and DMEM/F12 medium (10% fetal bovine serum, 100 U/ml penicillin G, 100 g/ml streptomycin added; Life Technologies, Grand Island, NY), 37°C, 5% CO-95 was maintained at % air condition.
<실시예 1> 용규 알코올 추출물의 자궁내막증 또는 이의 합병증 치료, 개선 또는 예방 효과 확인<Example 1> Confirmation of the effect of treatment, improvement or prevention of endometriosis or its complications of Yonggyu alcohol extract
1-1. 용규 알코올 추출물의 준비1-1. Preparation of Yonggyu Alcohol Extract
음건된 용규(Solanum nigrum, 산지: 경상북도 영천)을 대한민국 경동시장에서 구입하였고, 용규 전초를 400um 입자 사이즈가 되도록 분쇄하였다. 이와 같이 준비된 용규 10kg에 70%(v/v) 에탄올 수용액 100리터를 가하고 실온에서 24시간동안 침지하여 여액을 취하였고, 동일한 방법으로 1회 반복 추출하였다. 추출한 여액을 섭씨 60도에서 감압농축하고, 농축액을 동결건조하여 분말상태의 용규 70%에탄올 추출물 1.5kg을 얻었다.
Shade-dried yonggyu (Solanum nigrum, origin: Yeongcheon, Gyeongsangbuk-do) was purchased from the Gyeongdong market in Korea, and yonggyu outpost was pulverized to a particle size of 400um. 100 liters of 70% (v/v) ethanol aqueous solution was added to 10 kg of Yonggyu prepared in this way, and the filtrate was obtained by immersion at room temperature for 24 hours, and the extraction was repeated once in the same manner. The extracted filtrate was concentrated under reduced pressure at 60 degrees Celsius, and the concentrate was freeze-dried to obtain 1.5 kg of powdery Yonggyu 70% ethanol extract.
1-2. 세포이동 및 부착 관련 효소 발현 억제 효과 확인1-2. Confirmation of the effect of inhibiting the expression of enzymes related to cell migration and adhesion
용규 70부피% 에탄올 추출물이 자궁내막증 세포의 이동 및 침투에 중요한 역할을 하는 단백질 분해효소인 MMP-2 및 MMP-9의 mRNA 발현을 억제하는지 확인하기 위하여, 실시간 PCR (real-time PCR)을 수행하였다.To confirm that Yonggyu 70% ethanol extract by volume inhibits the mRNA expression of MMP-2 and MMP-9, proteolytic enzymes that play an important role in the migration and penetration of endometriosis cells, real-time PCR was performed. did.
구체적으로, 자궁내막증 세포인 12Z(epithelial endometriotic cells)에 용규 70부피% 에탄올 추출물 25 ug/mL, 50 ug/mL, 또는 100 ug/mL을 각각 24시간 처리한 후 12Z 세포에서 TRIzol(Invitrogen Canada, Burlington, ON, Canada)을 사용하여 총 RNA를 추출한 후, 역전사 중합효소연쇄반응(reverse transcription polymerase chain reaction, RT-PCR)을 실시하였다. 구체적으로, first-strand cDNA를 주형으로 하여, Thermal Cycler Dice Real Time PCR System(Takara, Japan)을 이용하여 섭씨 95도에서 5초 동안 변성, 섭씨 57도에서 10초 동안 어닐링(annealing), 섭씨 72도에서 20초 동안 연장을 수행하는 조건으로 45회 반복하여 SYBR Green real-time PCR을 수행하였다. 상기 SYBR Green real-time PCR에 사용된 MMP-2, MMP-9 및 보정용 GAPDH 프라이머들은 하기 [표 1]와 같으며, MMP-2, MMP-9의 Ct값의 평균은 트리플리케이트(triplicate) 측정으로 구해지며, GAPDH의 Ct값에 의해 보정되었다. 또한, 용규 추출물 미처리군은 대조군(control; CON)으로 하여, 추출물 처리를 제외하고 용규 추출물 처리군과 동일하게 처리하였다. 그 결과, 자궁내막증에서 발현되는 대표적인 단백질이며, 세포의 이동과 침투에 중요한 역할을 하는 내인성 MMP-2 및 MMP-9의 mRNA 발현을 용규 70% 에탄올 추출물이 억제함을 자궁내막증 세포인 12Z 세포에서 확인하였다(도 1, 도 2 참조).Specifically, 12Z (epithelial endometriotic cells), which are endometriosis cells, were treated with 25 ug/mL, 50 ug/mL, or 100 ug/mL of Yonggyu 70% ethanol extract for 24 hours, respectively, and then TRIzol (Invitrogen Canada, Burlington, ON, Canada) was used to extract total RNA, and then reverse transcription polymerase chain reaction (RT-PCR) was performed. Specifically, using the first-strand cDNA as a template, using the Thermal Cycler Dice Real Time PCR System (Takara, Japan), denatured at 95 degrees Celsius for 5 seconds, annealed at 57 degrees Celsius for 10 seconds, 72 degrees Celsius SYBR Green real-time PCR was performed by repeating 45 times under the condition of performing extension for 20 seconds in FIG. The MMP-2, MMP-9, and GAPDH primers for correction used in the SYBR Green real-time PCR are shown in Table 1 below, and the average of the Ct values of MMP-2 and MMP-9 is a triplicate. It was determined by measurement and corrected by the Ct value of GAPDH. In addition, the Yonggyu extract untreated group was treated as a control (control; CON), and was treated in the same manner as the Yonggyu extract treated group except for the extract treatment. As a result, it was found that Yonggyu 70% ethanol extract inhibited the mRNA expression of endogenous MMP-2 and MMP-9, which are representative proteins expressed in endometriosis and play an important role in cell migration and penetration, in 12Z cells, endometriosis cells. was confirmed (see FIGS. 1 and 2).
[표 1][Table 1]
도 1은 용규 알콜 추출물의 MMP-2 발현 억제 효과 확인 실험 결과를 나타낸 도이고, 도 2는 용규 알콜 추출물의 MMP-9 발현 억제 효과 확인 실험 결과를 나타낸 도로, 70부피% 에탄올 추출물 25ug/mL의 농도에서도, 효과적으로 MMP-2와 MMP-9 발현을 억제함을 알 수 있다.
1 is a diagram showing the experimental results for confirming the MMP-2 expression inhibitory effect of Yonggyu alcohol extract, Figure 2 is a road showing the experimental results confirming the MMP-9 expression inhibitory effect of the Yonggyu alcohol extract, 70% by volume ethanol extract 25ug/mL It can be seen that even at the concentration, it effectively inhibits the expression of MMP-2 and MMP-9.
1-3. 염증 및 통증 관련 효소 발현 억제 효과 확인1-3. Confirmation of the effect of inhibiting the expression of inflammation and pain-related enzymes
용규 70부피% 에탄올 추출물이 자궁내막증의 통증과 관련되어 있다고 알려진 COX-2의 mRNA 발현에 미치는 영향을 확인하기 위하여, 실시간 PCR (real-time PCR)을 수행하였다. 구체적으로, 자궁내막증 세포인 12Z (epithelial endometriotic cells)에 용규 70% 에탄올 추출물 25 ug/mL, 50 ug/mL, 또는 100 ug/mL 각각 24시간 처리한 후 12Z 세포에서 TRIzol (Invitrogen Canada, Burlington, ON, Canada)을 사용하여 총 RNA를 추출하여, 역전사 중합효소연쇄반응 (reverse transcription polymerase chain reaction, RT-PCR)을 실시하였다. 구체적으로, first-strand cDNA를 주형으로 하여, Thermal Cycler Dice Real Time PCR System (Takara, Japan)을 이용하여 섭씨 95도에서 5초 동안 변성, 섭씨 57도에서 10초 동안 어닐링 (annealing), 섭씨 72도에서 20초 동안 연장을 수행하는 조건으로 45회 반복하여 SYBR Green real-time PCR을 수행하였다. 상기 SYBR Green real-time PCR에 사용된 COX-2 프라이머, 보정용 GAPDH 프라이머들은 하기 [표 2]와 같으며, COX-2의 Ct값의 평균은 트리플리케이트(triplicate) 측정으로 구해지며, GAPDH의 Ct값에 의해 보정되었다. 또한, 용규 추출물 미처리군은 대조군(control; CON)으로 하여, 추출물 처리를 제외하고 용규 추출물 처리군과 동일하게 처리하였다.To confirm the effect of Yonggyu 70% ethanol extract by volume on the mRNA expression of COX-2, which is known to be associated with endometriosis pain, real-time PCR was performed. Specifically, 12Z (epithelial endometriotic cells), which are endometriosis cells, were treated with 25 ug/mL, 50 ug/mL, or 100 ug/mL of Yonggyu 70% ethanol extract for 24 hours, respectively, and then TRIzol (Invitrogen Canada, Burlington, ON, Canada) was used to extract total RNA, and reverse transcription polymerase chain reaction (RT-PCR) was performed. Specifically, using the first-strand cDNA as a template, using the Thermal Cycler Dice Real Time PCR System (Takara, Japan), denatured at 95 degrees Celsius for 5 seconds, annealed at 57 degrees Celsius for 10 seconds, 72 degrees Celsius SYBR Green real-time PCR was performed by repeating 45 times under the condition of performing extension for 20 seconds in FIG. The COX-2 primer and the GAPDH primer for correction used in the SYBR Green real-time PCR are as shown in Table 2 below, and the average of the Ct values of COX-2 is obtained by triplicate measurement, and the GAPDH Corrected by Ct value. In addition, the Yonggyu extract untreated group was treated as a control (control; CON), and was treated in the same manner as the Yonggyu extract treated group except for the extract treatment.
[표 2][Table 2]
그 결과, 자궁내막증 세포의 통증 관련 특징적 효소인 COX-2의 mRNA 발현을 용규 70% 에탄올 추출물이 억제함을 자궁내막증 세포인 12Z 세포에서 확인하였다(도 3 참조).As a result, it was confirmed in 12Z cells of endometriosis cells that the 70% ethanol extract of Yonggyu inhibited the mRNA expression of COX-2, a characteristic pain-related enzyme in endometriosis cells (see FIG. 3 ).
도 3은 용규 알콜 추출물의 COX-2 발현 억제 효과 확인 실험 결과를 나타낸 도로, 70부피% 에탄올 추출물이 25ug/mL의 농도에서도, 효과적으로 COX-2 발현을 억제함을 알 수 있다.
3 is a road showing the experimental results for confirming the COX-2 expression inhibition effect of the Yong-gyu alcohol extract, it can be seen that the 70 vol% ethanol extract effectively inhibits the COX-2 expression even at a concentration of 25 ug/mL.
1-4. 염증성 싸이토카인 발현 억제 효과 확인1-4. Confirmation of inhibitory effect on inflammatory cytokine expression
용규 70부피% 에탄올 추출물이 자궁내막증 세포에서 과발현되는 것으로 알려져 있는 염증성 사이토카인 mRNA 발현에 영향을 미치는지 확인하기 위하여, 실시간 PCR (real-time PCR)을 수행하였다.In order to determine whether Yonggyu 70% ethanol extract by volume affects the mRNA expression of inflammatory cytokines known to be overexpressed in endometriosis cells, real-time PCR was performed.
구체적으로, 자궁내막증 세포인 12Z (epithelial endometriotic cells)에 용규 70% 에탄올 추출물 25 ug/mL, 50 ug/mL, 또는 100 ug/mL 각각 24시간 처리한 후 12Z 세포에서 TRIzol (Invitrogen Canada, Burlington, ON, Canada)을 사용하여 총 RNA를 추출하여, 역전사 중합효소연쇄반응 (reverse transcription polymerase chain reaction, RT-PCR)을 실시하였다. 구체적으로, first-strand cDNA를 주형으로 하여, Thermal Cycler Dice Real Time PCR System (Takara, Japan)을 이용하여 섭씨 95도에서 5초 동안 변성, 섭씨 57도에서 10초 동안 어닐링 (annealing), 섭씨 72도에서 20초 동안 연장을 수행하는 조건으로 45회 반복하여 SYBR Green real-time PCR을 수행하였다. 상기 SYBR Green real-time PCR에 사용된 TNF-a 프라이머, MCP-1 프라이머 및 보정용 GAPDH 프라이머들은 하기 [표 3]와 같으며, TNF-a 및 MCP-1의 Ct값의 평균은 트리플리케이트(triplicate) 측정으로 구해지며, GAPDH의 Ct값에 의해 보정되었다. 또한, 용규 추출물 미처리군은 대조군(control; CON)으로 하여, 추출물 처리를 제외하고 용규 추출물 처리군과 동일하게 처리하였다.Specifically, 12Z (epithelial endometriotic cells), which are endometriosis cells, were treated with 25 ug/mL, 50 ug/mL, or 100 ug/mL of Yonggyu 70% ethanol extract for 24 hours, respectively, and then TRIzol (Invitrogen Canada, Burlington, ON, Canada) was used to extract total RNA, and reverse transcription polymerase chain reaction (RT-PCR) was performed. Specifically, using the first-strand cDNA as a template, using the Thermal Cycler Dice Real Time PCR System (Takara, Japan), denatured at 95 degrees Celsius for 5 seconds, annealed at 57 degrees Celsius for 10 seconds, 72 degrees Celsius SYBR Green real-time PCR was performed by repeating 45 times under the condition of performing extension for 20 seconds in FIG. The TNF-a primer, the MCP-1 primer, and the GAPDH primer for correction used in the SYBR Green real-time PCR are as shown in Table 3 below, and the average of the Ct values of TNF-a and MCP-1 is triplicate ( triplicate) and was corrected by the Ct value of GAPDH. In addition, the Yonggyu extract untreated group was treated as a control (control; CON), and was treated in the same manner as the Yonggyu extract treated group except for the extract treatment.
[표 3][Table 3]
그 결과, 자궁내막증 세포의 염증 관련 사이토카인인 TNF-알파 및 MCP-1의 mRNA 발현을 용규 70% 에탄올 추출물이 억제함을 자궁내막증 세포인 12Z 세포에서 확인하였다(도 4, 도 5 참조).As a result, it was confirmed in 12Z cells, which are endometriosis cells, that the 70% Yonggyu extract inhibited the mRNA expression of TNF-alpha and MCP-1, which are inflammation-related cytokines, in endometriosis cells (see FIGS. 4 and 5).
도 4는 용규 알콜 추출물의 TNF-알파 발현 억제 효과 확인 실험 결과를 나타낸 도이고, 도 5는 용규 알콜 추출물의 MCP-1 발현 억제 효과 확인 실험 결과를 나타낸 도로, 70부피% 에탄올 추출물이 25ug/mL의 농도에서도, 효과적으로 TNF-알파 및 MCP-1 발현을 억제함을 알 수 있다.
Figure 4 is a diagram showing the experimental results of confirming the TNF-alpha expression inhibitory effect of Yonggyu alcohol extract, Figure 5 is a road showing the experimental results confirming the MCP-1 expression inhibitory effect of Yonggyu alcohol extract, 70% by volume ethanol extract 25ug / mL Even at a concentration of, it can be seen that it effectively inhibits TNF-alpha and MCP-1 expression.
1-5. 자궁내막증 세포 부착 및 착상 억제 효과 확인1-5. Confirmation of endometriosis cell adhesion and implantation inhibition effect
용규 70% 에탄올 추출물이 자궁내막증 세포의 부착 및 착상에 미치는 영향을 확인하기 위하여 부착 분석 (attachment assay)을 3번 반복 수행하였다. In order to confirm the effect of Yonggyu 70% ethanol extract on the adhesion and implantation of endometriosis cells, the attachment assay was repeated three times.
구체적으로, 자궁내막증의 특징인 자궁내막증 세포가 복막 복강주위의 장기에 부착 및 착상하는 현상을 용규 70% 에탄올 추출물이 억제하는지 확인하기 위해, 인간 복막 중피 세포(Human peritoneal methothelial cells)인 Met-5A 세포 (American Type Culture Collection, ATCC)를 10%의 FBS, 100 U/ml의 페니실린 G 및 100 g/ml의 스트렙토마이신 (Life Technologies, Grand Island, NY, U.S.A.), 및 400 nM의 hydrocortisone (Sigma Chemical Co, St. Louis, MO, U.S.A.)가 포함된 199 배지에서 5 % CO2 인큐베이터로 배양하였다. 상기 Met-5A 세포를 96-웰 플레이트에 분주하였다. Cell tracker green CMFDA로 표지한 자궁내막증 세포주인 12Z 세포 (2 X 103 개/well)를 50 ug/mL, 100 ug/mL, 또는 200 ug/mL 의 용규 70% 에탄올 추출물을 함유한 DMEM/F12배지와 섞은 후, 미리 분주해놓은 Met-5A 세포에 첨가하였다. 섞인 상기의 세포들은 섭씨37도의 5% CO2-인큐베이터에서 한 시간 동안 배양한 후, 플레이트를 뒤집고 칼슘 및 마그네슘이 들어있는 포스페이트-버퍼 용액 (phosphate-buffered solution)으로 씻었다. 부착된 세포들을 490 nm의 파장으로 분석하여, 보정용 대조군을 100%으로 놓고 부착된 세포들의 퍼센트를 계산하였다. 또한, 용규 추출물 미처리군은 대조군(control; CON)으로 하여, 추출물 처리를 제외하고 용규 추출물 처리군과 동일하게 처리하였다.Specifically, in order to confirm whether the Yonggyu 70% ethanol extract inhibits the attachment and implantation of endometriosis cells characteristic of endometriosis to organs around the peritoneum, Met-5A, a human peritoneal mesothelial cell Cells (American Type Culture Collection, ATCC) were cultured with 10% FBS, 100 U/ml penicillin G and 100 g/ml streptomycin (Life Technologies, Grand Island, NY, USA), and 400 nM hydrocortisone (Sigma Chemical). Co, St. Louis, MO, USA) in 199 medium containing 5% CO2 incubator. The Met-5A cells were seeded in 96-well plates. Cell tracker green CMFDA-labeled endometriosis cell line 12Z cells (2
그 결과, 용규 70% 에탄올 추출물을 처리한 자궁내막증 세포 12Z가, 복막 및 복강 주위 장기의 외부를 둘러싼 Met-5A 세포에 부착 억제하는 것을 확인하였다(도 6 참조).As a result, it was confirmed that endometriosis cells 12Z treated with Yonggyu 70% ethanol extract inhibited adhesion to Met-5A cells surrounding the peritoneum and the organs around the abdominal cavity (see FIG. 6 ).
도 6은 용규 알콜 추출물의 자궁내막증 세포 부착 및 착상 억제 효과 확인 실험 결과를 나타낸 도로, 70부피% 에탄올 추출물이 50ug/mL의 농도에서도, 효과적으로 세포 부착을 억제함을 알 수 있다.
6 is a road showing the test results for confirming the effect of Yonggyu alcohol extract to inhibit endometriosis cell adhesion and implantation, and it can be seen that 70% by volume ethanol extract effectively inhibits cell adhesion even at a concentration of 50 ug/mL.
1-6. 자궁내막증 세포 이동 억제 효과 확인1-6. Confirmation of endometriosis cell migration inhibitory effect
용규 70% 에탄올 추출물이 자궁내막증 세포의 이동에 미치는 영향을 확인하기 위하여 트랜스웰-이동 분석 (transwell-migration assay)을 3번 반복 수행하였다. 구체적으로, 24웰-트랜스웰 플레이트 (8 um pore size)에 자궁내막증 세포인 12Z 세포를 트랜스웰 플레이트의 윗 부분 (upper part)에 각각 분주한 후, 25 ug/mL, 50 ug/mL, 및 100 ug/mL 의 용규 70% 에탄올 추출물을 처리하였다. 용규 70% 에탄올 추출물 처리 후, 트랜스웰 바깥쪽면의 막으로 이동한 세포들을 메탄올로 고정하여, 0.5%의 크리스탈 바이올렛으로 10분 동안 염색하였다. 상기의 트랜스웰 바깥쪽 면으로 이동한 세포들을 200 X 배율의 현미경으로 관찰하여 개수하여 그래프화 하였다. 또한, 용규 추출물 미처리군은 대조군(control; CON)으로 하여, 추출물 처리를 제외하고 용규 추출물 처리군과 동일하게 처리하였다. 그 결과, 용규 70% 에탄올 추출물을 처리한 자궁내막증 세포 12Z의 이동이 유의적으로 억제됨을 확인하였다(도 7 참조).In order to confirm the effect of Yonggyu 70% ethanol extract on the migration of endometriosis cells, the transwell-migration assay was repeated three times. Specifically, after each dispensing of 12Z cells, which are endometriosis cells, into the upper part of the transwell plate in a 24-well-transwell plate (8 um pore size), 25 ug/mL, 50 ug/mL, and 100 ug/mL of Yonggyu 70% ethanol extract was treated. After treatment with Yonggyu 70% ethanol extract, the cells that migrated to the membrane on the outer surface of the transwell were fixed with methanol and stained with 0.5% crystal violet for 10 minutes. Cells that migrated to the outer surface of the transwell were observed under a microscope at 200 X magnification, counted and graphed. In addition, the Yonggyu extract untreated group was treated as a control (control; CON), and was treated in the same manner as the Yonggyu extract treated group except for the extract treatment. As a result, it was confirmed that the migration of endometriosis cells 12Z treated with Yonggyu 70% ethanol extract was significantly inhibited (see FIG. 7 ).
도 7은 용규 알콜 추출물의 자궁내막증 세포 이동 억제 효과 확인 실험 결과를 나타낸 도로, 70부피% 에탄올 추출물이 25ug/mL의 농도에서도, 효과적으로 세포 부착을 억제함을 알 수 있다.
7 is a road showing the test results for confirming the endometriosis cell migration inhibitory effect of Yonggyu alcohol extract, it can be seen that 70% by volume ethanol extract effectively inhibits cell adhesion even at a concentration of 25ug/mL.
1-7. 복강내투여(Intraperitoneal injection)유도모델에서의 효과 확인1-7. Confirmation of effectiveness in intraperitoneal injection induction model
용규 70% 에탄올 추출물이 Intraperitoneal injection 자궁내막증 유도 모델에서의 효과를 확인하기 위하여 평가를 수행하였다. An evaluation was performed to confirm the effect of Yonggyu 70% ethanol extract in the intraperitoneal injection endometriosis induction model.
자궁내막증 유도는 Donor mouse에서 이식할 자궁을 적출하여 잘게 조각한 후 recipient mouse의 복강내로 spreading하여 유도하였고, 약물투여는 하기 [표 4]와 같으며, 용규 70% 에탄올 추출물 125 mg/kg, 250mg/kg, 500 mg/kg을 경구로 5주간 투여하였다. 자궁내막증 유도된 동물모델을 희생시켜 복부를 개복하여 복막과 복강 내 (소장, 대장, 위, 간, 방광, 난소 등)의 implantation 수를 확인하였다.
Induction of endometriosis was induced by removing the uterus to be transplanted from the donor mouse, sculpting it, and spreading it into the abdominal cavity of the recipient mouse. /kg, 500 mg/kg was orally administered for 5 weeks. After sacrificing an endometriosis-induced animal model, the abdomen was opened and the number of implantations in the peritoneum and intraperitoneal cavity (small intestine, large intestine, stomach, liver, bladder, ovary, etc.) was confirmed.
[표 4][Table 4]
또한, 비클(Vehicle) 투여군은, 추출물 대신 10% Tween 80을 200ul 투여한 것을 제외하고 용규 추출물 투여군과 동일하게 처리하였다. 또한, 양성 대조군은 용규 추출물 대신 dienogest를 0.3mg/kg 투여한 것을 제외하고, 용규 추출물 투여군과 동일하게 처리하였다.In addition, the vehicle administration group was treated the same as the Yonggyu extract administration group except that 200ul of 10% Tween 80 was administered instead of the extract. In addition, the positive control group was treated in the same manner as the Yonggyu extract administration group, except that 0.3 mg/kg of dienogest was administered instead of the Yonggyu extract.
그 결과, 용규 70% 에탄올 추출물이 유의적으로 자궁내막증 조직형성 억제 효과가 있는 것으로 확인하였다(도 8 참조).As a result, it was confirmed that the 70% ethanol extract of Yonggyu had a significant inhibitory effect on endometriosis tissue formation (see FIG. 8 ).
도 8은 용규 알콜 추출물의 복강내투여모델에서 효과 확인 실험 결과를 나타낸 도로, 70부피% 에탄올 추출물이 125mg/kg에서도, 효과적으로 자궁내막증 조직형성 억제 효과를 나타냄을 알 수 있다.
8 is a road showing the experimental results for confirming the effect in the intraperitoneal administration model of the Yonggyu alcohol extract, it can be seen that the 70% by volume ethanol extract effectively exhibits the endometriosis tissue formation inhibitory effect even at 125 mg/kg.
1-8. 수술 유도 모델(Surgical induced model)에서의 효과 확인1-8. Confirmation of effectiveness in surgical induced model
용규 70% 에탄올 추출물이 Surgical induced 자궁내막증 유도 모델에서의 효과를 확인하기 위한 평가를 수행하였다. An evaluation was performed to confirm the effect of Yonggyu 70% ethanol extract in a Surgical-induced endometriosis induction model.
자궁내막증 유도는 SD rat (female)의 왼쪽 자궁을 7 mm길이로 절단 후 내막 부분이 복막에 맞닿게 양쪽 끝을 silkam으로 봉합하는 방법으로 자궁내막세포를 이식하여 유도하였고, 약물투여는 하기 [표 5]와 같으며, 용규 70% 에탄올 추출물 100 mg/kg, 300mg/kg, 500 mg/kg을 경구로 4주간 투여하였다. 자궁내막증 유도된 동물모델을 희생시켜 복부를 개복하여 자궁내막증 세포를 이식한 복막에서의 implant 크기{염증 부위의 크기 (가로*세로*높이)}를 확인하였다.
Endometriosis was induced by transplanting endometrial cells by cutting the left uterus of an SD rat (female) to a length of 7 mm and sealing both ends with silkam so that the endometrial part was in contact with the peritoneum. 5], and Yonggyu 70
[표 5][Table 5]
또한, 비클(Vehicle) 투여군은, 추출물 대신 추출물을 녹이기 위한 용매를 체중에 비례하여 투여한 것을 제외하고 용규 추출물 투여군과 동일하게 처리하였다. 추출물은 10% Tween 80으로 녹였다. 또한, 양성 대조군(Positive control)은 용규 추출물 대신 dienogest를 0.3mg/kg 투여한 것을 제외하고, 용규 추출물 투여군과 동일하게 처리하였다.In addition, the vehicle administration group was treated in the same manner as the Yonggyu extract administration group except that a solvent for dissolving the extract was administered in proportion to the body weight instead of the extract. The extract was dissolved with 10% Tween 80. In addition, the positive control group was treated in the same manner as the Yonggyu extract administration group, except that 0.3 mg/kg of dienogest was administered instead of the Yonggyu extract.
그 결과, 용규 70% 에탄올 추출물이 자궁내막증 조직형성 억제 효과가 있는 것으로 확인하였다(도 9 참조).As a result, it was confirmed that the Yonggyu 70% ethanol extract had the effect of inhibiting endometriosis tissue formation (see FIG. 9 ).
도 9는 용규 알콜 추출물의 수술유도모델에서 효과 확인 실험 결과를 나타낸 도로, 70부피% 에탄올 추출물이 100mg/kg에서도, 효과적으로 자궁내막증 조직형성 억제 효과를 나타냄을 알 수 있다.9 is a road showing the experimental results for confirming the effect of the yonggyu alcohol extract in the surgical induction model, it can be seen that the 70% by volume ethanol extract effectively exhibits the effect of inhibiting endometriosis tissue formation even at 100 mg/kg.
이와 같은 결과로부터, 용규 알코올 추출물이 자궁내막증 또는 이의 합병증을 치료, 개선 또는 예방 효과를 확인할 수 있다.
From these results, it can be confirmed that Yonggyu alcohol extract treats, improves or prevents endometriosis or its complications.
<실시예 2> 용규 부탄올분획 추출물의 자궁내막증 및 합병증 치료, 개선 또는 예방 효과 확인<Example 2> Confirmation of effect of treatment, improvement or prevention of endometriosis and complications of Yonggyu butanol fraction extract
2-1. 용규 부탄올분획 추출물의 준비2-1. Preparation of Yonggyu butanol fraction extract
1-1.과 동일한 방법으로 준비한 70%에탄올 추출물 1kg의 계통적인 분획을 통하여 노말헥산 분획, 디클로로메탄 분획, 에틸아세테이트 분획, 부탄올 분획, 그리고 물층으로 나누는 방법에 의해, 용규 부탄올분획 추출물을 준비하였다. 구체적으로, 분말상태의 추출물 1kg에 물 4L를 가하고, 물이 가해진 알콜 추출물을 n-헥산 20L로 분획추출하고, n-헥산 분획을 따로 취하고, 남은 수층을 디클로로메탄 20L로 분획추출하여 디클로로메탄 분획을 따로 취하고, 남은 수층을 에틸아세테이트 40L로 분획추출하여, 에틸아세테이트 분획을 따로 취하고, 남은 수층을 부탄올 40L로 분획추출하여, 부탄올 분획을 얻고, 이와 같은 과정에 의해 얻어진 부탄올 분획을 감압농축하고, 농축액을 동결건조하여 분말상태의 용규 70% 에탄올 추출물 중 부탄올 분획 추출물 141g을 얻었다. 또한, 부탄올 분획추출 후 남은 물층을 따로 얻었다.
Through the systematic fractionation of 1 kg of the 70% ethanol extract prepared in the same manner as in 1-1., the extract of the Yonggyu butanol fraction was prepared by dividing it into a normal hexane fraction, a dichloromethane fraction, an ethyl acetate fraction, a butanol fraction, and an aqueous layer. . Specifically, 4L of water was added to 1kg of the powdery extract, the alcohol extract to which water was added was fractionally extracted with 20L of n-hexane, the n-hexane fraction was separately taken, and the remaining aqueous layer was fractionated with 20L of dichloromethane to fractionate dichloromethane. was separately taken, the remaining aqueous layer was fractionally extracted with 40 L of ethyl acetate, the ethyl acetate fraction was separated, and the remaining aqueous layer was fractionally extracted with 40 L of butanol to obtain a butanol fraction, and the butanol fraction obtained by this process was concentrated under reduced pressure, The concentrate was freeze-dried to obtain 141 g of the butanol fraction extract in the powdery Yonggyu 70% ethanol extract. In addition, the water layer remaining after the butanol fractionation was obtained separately.
2-2. 세포이동 및 부착 관련 효소 발현 억제 효과 확인2-2. Confirmation of the effect of inhibiting the expression of enzymes related to cell migration and adhesion
용규 70부피% 에탄올 추출물 25 ug/mL, 50 ug/mL, 또는 100 ug/mL 대신, 2-1.의 용규 부탄올분획 추출물 0.5 ug/mL, 1 ug/mL, 또는 2 ug/mL을 사용한 것을 제외하고, 실시예 1-2.과 동일하게 실시하였다.Instead of 25 ug/mL, 50 ug/mL, or 100 ug/mL of Yonggyu's 70 vol% ethanol extract, 0.5 ug/mL, 1 ug/mL, or 2 ug/mL of Yonggyu's butanol fraction extract from 2-1 was used. Except, it was carried out in the same manner as in Example 1-2.
그 결과, 자궁내막증에서 발현되는 대표적인 단백질이며, 세포의 이동과 침투에 중요한 역할을 하는 내인성 MMP-2 및 MMP-9의 mRNA 발현을 용규 70% 에탄올 추출물의 부탄올분획추출물이 억제함을 자궁내막증 세포인 12Z 세포에서 확인하였다(도 10, 도 11 참조).As a result, it was found that the butanol fraction extract of Yonggyu 70% ethanol extract inhibited the mRNA expression of endogenous MMP-2 and MMP-9, which are representative proteins expressed in endometriosis and play an important role in cell migration and penetration, in endometriosis cells. It was confirmed in phosphorus 12Z cells (see FIGS. 10 and 11).
도 10은 용규 부탄올분획추출물의 MMP-2 발현 억제 효과 확인 실험 결과를 나타낸 도이고, 도 11은 용규 부탄올분획추출물의 MMP-9 발현 억제 효과 확인 실험 결과를 나타낸 도로, 용규 부탄올분획추출물 0.5ug/mL의 농도에서도, 효과적으로 MMP-2와 MMP-9 발현을 억제함을 알 수 있다.
10 is a diagram showing the results of the MMP-2 expression inhibitory effect of the Yonggyu butanol fraction extract, and FIG. 11 is the road showing the MMP-9 expression inhibitory effect of the Yonggyu butanol fraction extract 0.5ug/ It can be seen that even at a concentration of mL, it effectively inhibits MMP-2 and MMP-9 expression.
2-3. 염증 및 통증 관련 효소 발현 억제 효과 확인2-3. Confirmation of the effect of inhibiting the expression of inflammation and pain-related enzymes
용규 70% 에탄올 추출물 25 ug/mL, 50 ug/mL, 또는 100 ug/mL 대신, 2-1.의 용규 부탄올분획 추출물 0.5 ug/mL, 1 ug/mL, 또는 2 ug/mL을 사용한 것을 제외하고, 실시예 1-3.과 동일하게 실시하였다.Instead of 25 ug/mL, 50 ug/mL, or 100 ug/mL of Yonggyu's 70% ethanol extract, 0.5 ug/mL, 1 ug/mL, or 2 ug/mL of Yonggyu's butanol fraction extract from 2-1 was used and carried out in the same manner as in Example 1-3.
그 결과, 자궁내막증 세포의 통증 관련 특징적 효소인 COX-2의 mRNA 발현을 용규 부탄올분획 추출물이 억제함을 자궁내막증 세포인 12Z 세포에서 확인하였다(도 12 참조).As a result, it was confirmed in 12Z cells, which are endometriosis cells, that the Yonggyu butanol fraction extract inhibited the mRNA expression of COX-2, a characteristic pain-related enzyme in endometriosis cells (see FIG. 12 ).
도 12는 용규 부탄올분획추출물의 COX-2 발현 억제 효과 확인 실험 결과를 나타낸 도로, 용규 부탄올분획추출물이 0.5ug/mL의 농도에서도, 효과적으로 COX-2 발현을 억제함을 알 수 있다.
12 is a road showing the experimental results for confirming the COX-2 expression inhibition effect of the Yonggyu butanol fraction extract, it can be seen that the Yonggyu butanol fraction extract effectively inhibits the expression of COX-2 even at a concentration of 0.5ug/mL.
2-4. 염증성 싸이토카인 발현 억제 효과 확인2-4. Confirmation of inhibitory effect on inflammatory cytokine expression
용규 70% 에탄올 추출물 25 ug/mL, 50 ug/mL, 또는 100 ug/mL 대신, 2-1.의 용규 부탄올분획 추출물 0.5 ug/mL, 1 ug/mL, 또는 2 ug/mL을 사용한 것을 제외하고, 실시예 1-4.과 동일하게 실시하였다.Instead of 25 ug/mL, 50 ug/mL, or 100 ug/mL of Yonggyu's 70% ethanol extract, 0.5 ug/mL, 1 ug/mL, or 2 ug/mL of Yonggyu's butanol fraction extract from 2-1 was used and carried out in the same manner as in Examples 1-4.
그 결과, 자궁내막증 세포의 염증 관련 사이토카인인 TNF-알파 및 MCP-1의 mRNA 발현을 용규 부탄올분획 추출물이 억제함을 자궁내막증 세포인 12Z 세포에서 확인하였다(도 13, 도 14 참조).As a result, it was confirmed that the Yonggyu butanol fraction extract inhibited the mRNA expression of TNF-alpha and MCP-1, which are inflammation-related cytokines in endometriosis cells, in 12Z cells, which are endometriosis cells (see FIGS. 13 and 14).
도 13은 용규 부탄올분획 추출물의 TNF-알파 발현 억제 효과 확인 실험 결과를 나타낸 도이고, 도 14는 용규 부탄올분획 추출물의 MCP-1 발현 억제 효과 확인 실험 결과를 나타낸 도로, 용규 부탄올분획 추출물이 0.5ug/mL의 농도에서도, 효과적으로 TNF-알파 및 MCP-1 발현을 억제함을 알 수 있다.
13 is a diagram showing the experimental results for confirming the TNF-alpha expression inhibitory effect of the Yonggyu butanol fraction extract, and FIG. 14 is a road showing the experimental results for confirming the MCP-1 expression inhibitory effect of the Yonggyu butanol fraction extract. 0.5ug of the Yonggyu butanol fraction extract Even at a concentration of /mL, it can be seen that it effectively inhibits TNF-alpha and MCP-1 expression.
2-5. 자궁내막증 세포 부착 및 착상 억제 효과 확인2-5. Confirmation of endometriosis cell adhesion and implantation inhibition effect
50 ug/mL, 100 ug/mL, 또는 200 ug/mL 의 용규 70% 에탄올 추출물 대신, 2-1.의 용규 부탄올분획 추출물 0.5 ug/mL, 1 ug/mL, 또는 2 ug/mL을 사용한 것을 제외하고, 실시예 1-5.과 동일하게 실시하였다.Instead of 50 ug/mL, 100 ug/mL, or 200 ug/mL of Yonggyu 70% ethanol extract, 0.5 ug/mL, 1 ug/mL, or 2 ug/mL of Yonggyu butanol fraction extract from 2-1. Except that, it was carried out in the same manner as in Examples 1-5.
그 결과, 용규 부탄올분획 추출물을 처리한 자궁내막증 세포 12Z가, 복막 및 복강 주위 장기의 외부를 둘러싼 Met-5A 세포에 부착 억제하는 것을 확인하였다(도 15 참조).As a result, it was confirmed that the endometriosis cells 12Z treated with the Yonggyu butanol fraction extract inhibited adhesion to the Met-5A cells surrounding the peritoneum and the organs around the abdominal cavity (see FIG. 15 ).
도 15는 용규 부탄올분획 추출물의 자궁내막증 세포 부착 및 착상 억제 효과 확인 실험 결과를 나타낸 도로, 용규부탄올분획 추출물이 낮은 농도에서도, 효과적으로 세포 부착을 억제함을 알 수 있다.15 is a road showing the test results for confirming the effect of inhibiting endometriosis cell adhesion and implantation of the Yonggyu butanol fraction extract, it can be seen that the Yonggyu butanol fraction extract effectively inhibits cell adhesion even at a low concentration.
이와 같은 결과로부터, 용규 부탄올분획 추출물이 자궁내막증 또는 이의 합병증을 치료, 개선 또는 예방하는 효과가 있음을 확인할 수 있다.
From these results, it can be confirmed that the Yonggyu butanol fraction extract is effective in treating, improving or preventing endometriosis or its complications.
<실시예 3> 용규 분획 추출물의 자궁내막증 및 합병증 치료, 개선 또는 예방 효과 확인<Example 3> Confirmation of effect of treatment, improvement or prevention of endometriosis and complications of Yonggyu fraction extract
3-1. 용규 분획 추출물의 준비3-1. Preparation of Yonggyu fraction extract
1-2.의 분획 추출법과 동일한 방법으로 얻어진, 노말헥산 분획, 디클로로메탄 분획, 에틸아세테이트 분획, 및 부탄올 분획추출 후 남은 물층 각각을 감압농축하고, 농축액을 동결건조하여 용규 70% 에탄올 추출물 중 노말헥산 분획 추출물, 디클로로메탄 분획 추출물, 에틸아세테이트 분획 추출물, 및 물층 추출물을 각각 분말상태로 105g, 16g, 36g, 477g 얻었다.
The normal hexane fraction, dichloromethane fraction, ethyl acetate fraction, and butanol fraction obtained in the same manner as in the fractional extraction method in 1-2. were concentrated under reduced pressure, respectively, and the concentrate was freeze-dried to obtain normal in the 70% ethanol extract of Yonggyu. 105 g, 16 g, 36 g, and 477 g of a hexane fractional extract, a dichloromethane fractional extract, an ethyl acetate fractional extract, and an aqueous layer extract were obtained in powder form, respectively.
3-2. 세포이동 및 부착 관련 효소 발현 억제 효과 확인3-2. Confirmation of the effect of inhibiting the expression of enzymes related to cell migration and adhesion
용규 부탄올분획 추출물 대신, 표 6 또는 표 7에 기재된 분획추출물을 사용한 것을 제외하고, 실시예 2-2.과 동일하게 실시하였다.It was carried out in the same manner as in Example 2-2., except that the fractional extracts listed in Table 6 or 7 were used instead of the Yonggyu butanol fraction extract.
그 결과를 표 6과 표 7에 나타내었으며, 표 6은 MMP-2에 대한 결과이고, 표 7은 MMP-9에 대한 결과를 나타낸다.The results are shown in Tables 6 and 7, Table 6 shows the results for MMP-2, and Table 7 shows the results for MMP-9.
[표 6][Table 6]
[표 7][Table 7]
그 결과, 각각의 분획 추출물이 MMP-2 및/또는 MMP-9 발현을 억제함을 알 수 있다.
As a result, it can be seen that each fractional extract inhibits MMP-2 and/or MMP-9 expression.
3-3. 염증 및 통증 관련 효소 발현 억제 효과 확인3-3. Confirmation of the effect of inhibiting the expression of inflammation and pain-related enzymes
용규 부탄올분획 추출물 대신, 표 8에 기재된 분획추출물을 사용한 것을 제외하고, 실시예 2-3.과 동일하게 실시하였다.It was carried out in the same manner as in Example 2-3., except that the fractionated extract shown in Table 8 was used instead of the Yonggyu butanol fraction extract.
그 결과를 표 8에 나타내었다. 그 결과, 자궁내막증 세포의 통증 관련 특징적 효소인 COX-2의 mRNA 발현을 각 분획 추출물이 억제함을 알 수 있다.
The results are shown in Table 8. As a result, it can be seen that each fraction extract inhibits the mRNA expression of COX-2, a pain-related characteristic enzyme in endometriosis cells.
[표 8][Table 8]
3-4. 염증성 싸이토카인 발현 억제 효과 확인3-4. Confirmation of inhibitory effect on inflammatory cytokine expression
용규 부탄올분획 추출물 대신, 표 9 또는 표 10에 기재된 분획 추출물을 사용한 것을 제외하고, 실시예 2-4.과 동일하게 실시하였다.It was carried out in the same manner as in Example 2-4, except that the fractional extracts shown in Table 9 or Table 10 were used instead of the Yonggyu butanol fraction extract.
그 결과를 표 9와 표 10에 나타내었으며, 표 9는 TNF-알파에 대한 결과이고, 표 10의 MCP-1에 대한 결과이다. 그 결과, 자궁내막증 세포의 염증 관련 사이토카인인 TNF-알파 및 MCP-1의 mRNA 발현을 각각의 분획 추출물이 억제함을 알 수 있다.The results are shown in Tables 9 and 10, and Table 9 is the results for TNF-alpha, and the results for MCP-1 in Table 10. As a result, it can be seen that each fraction extract inhibits the mRNA expression of TNF-alpha and MCP-1, which are inflammation-related cytokines in endometriosis cells.
[표 9][Table 9]
[표 10][Table 10]
이와 같은 결과로부터, 용규 분획 추출물이 자궁내막증 및 합병증의 치료, 개선 또는 예방에 효과적임을 확인할 수 있다.
From these results, it can be confirmed that the Yonggyu fraction extract is effective in the treatment, improvement or prevention of endometriosis and complications.
<실시예 4> 용규 물추출물의 자궁내막증 및 합병증 치료, 개선 또는 예방 효과 확인<Example 4> Confirmation of effect of treatment, improvement or prevention of endometriosis and complications of Yonggyu water extract
4-1. 용규 물추출물의 준비4-1. Preparation of Yonggyu Water Extract
음건된 용규(Solanum nigrum, 산지: 경상북도 영천)을 대한민국 경동시장에서 구입하였고, 용규 전초를 분쇄하였다. 이와 같이 준비된 용규 500g에 3차 증류수 1리터를 넣고 3시간동안 약탕기를 이용하여 추출하였다. 추출액을 여과지를 이용하여 여과한 후 48시간 동결건조하여 분말상태의 용규 물 추출물을 얻었다.
Shade-dried yonggyu (Solanum nigrum, origin: Yeongcheon, Gyeongsangbuk-do) was purchased at the Gyeongdong market in Korea, and yonggyu outpost was crushed. 1 liter of tertiary distilled water was added to 500 g of Yonggyu prepared in this way, and extracted using a yaktang for 3 hours. The extract was filtered using filter paper and then freeze-dried for 48 hours to obtain a powdery Yonggyu water extract.
4-2. 세포이동 및 부착 관련 효소 발현 억제 효과 확인4-2. Confirmation of the effect of inhibiting the expression of enzymes related to cell migration and adhesion
용규 70부피% 에탄올 추출물 25 ug/mL, 50 ug/mL, 또는 100 ug/mL 대신, 4-1.의 용규 물추출물 100 ug/mL을 사용한 것을 제외하고, 실시예 1-2.과 동일하게 실시하였다.Instead of 25 ug/mL, 50 ug/mL, or 100 ug/mL of Yonggyu 70 vol% ethanol extract, 100 ug/mL of Yonggyu water extract of 4-1. was used, as in Example 1-2. carried out.
그 결과, 자궁내막증에서 발현되는 대표적인 단백질이며, 세포의 이동과 침투에 중요한 역할을 하는 내인성 MMP-2의 mRNA 발현을 용규 물추출물이 억제함을 자궁내막증 세포인 12Z 세포에서 확인하였다(도 16 참조).As a result, it was confirmed in 12Z cells, which are endometriosis cells, that the water extract inhibited the mRNA expression of endogenous MMP-2, which is a representative protein expressed in endometriosis and plays an important role in cell migration and penetration (see Fig. 16). ).
도 16은 용규 물추출물의 MMP-2 발현 억제 효과 확인 실험 결과를 나타낸 도로, 용규 물추출물이 효과적으로 MMP-2 발현을 억제함을 알 수 있다.
16 is a road showing the experimental results for confirming the MMP-2 expression inhibition effect of the Yonggyu water extract, it can be seen that the Yonggyu water extract effectively inhibits the MMP-2 expression.
4-3. 염증 및 통증 관련 효소 발현 억제 효과 확인4-3. Confirmation of the effect of inhibiting the expression of inflammation and pain-related enzymes
용규 70% 에탄올 추출물 25 ug/mL, 50 ug/mL, 또는 100 ug/mL 대신, 4-1.의 용규 물추출물 100 ug/mL을 사용한 것을 제외하고, 실시예 1-3.과 동일하게 실시하였다.Instead of 25 ug/mL, 50 ug/mL, or 100 ug/mL of Yonggyu's 70% ethanol extract, 100 ug/mL of Yonggyu's water extract was used in the same manner as in Example 1-3. did.
그 결과, 자궁내막증 세포의 통증 관련 특징적 효소인 COX-2의 mRNA 발현을 용규 물추출물이 억제함을 자궁내막증 세포인 12Z 세포에서 확인하였다(도 17 참조).As a result, it was confirmed in 12Z cells, which are endometriosis cells, that the water extract inhibited the mRNA expression of COX-2, a characteristic pain-related enzyme in endometriosis cells (see FIG. 17 ).
도 17은 용규 물추출물의 COX-2 발현 억제 효과 확인 실험 결과를 나타낸 도로, 용규 물추출물이 효과적으로 COX-2 발현을 억제함을 알 수 있다.
17 is a road showing the experimental results for confirming the COX-2 expression inhibition effect of the Yonggyu water extract, it can be seen that the Yonggyu water extract effectively inhibits the COX-2 expression.
4-4. 염증성 싸이토카인 발현 억제 효과 확인4-4. Confirmation of inhibitory effect on inflammatory cytokine expression
용규 70% 에탄올 추출물 25 ug/mL, 50 ug/mL, 또는 100 ug/mL 대신, 4-1.의 용규 물추출물 100 ug/mL을 사용한 것을 제외하고, 실시예 1-4.과 동일하게 실시하였다.Instead of 25 ug/mL, 50 ug/mL, or 100 ug/mL of Yonggyu 70% ethanol extract, the same procedure as in Example 1-4, except that 100 ug/mL of Yonggyu water extract of 4-1 was used did.
그 결과, 자궁내막증 세포의 염증 관련 사이토카인인 TNF-알파 및 MCP-1의 mRNA 발현을 용규 물추출물이 억제함을 자궁내막증 세포인 12Z 세포에서 확인하였다(도 18, 도 19 참조).As a result, it was confirmed in 12Z cells, which are endometriosis cells, that the water extract inhibited the mRNA expression of TNF-alpha and MCP-1, which are inflammation-related cytokines in endometriosis cells (see FIGS. 18 and 19).
도 18은 용규 물추출물의 TNF-알파 발현 억제 효과 확인 실험 결과를 나타낸 도이고, 도 19는 용규 물추출물의 MCP-1 발현 억제 효과 확인 실험 결과를 나타낸 도로, 용규 물추출물이 효과적으로 TNF-알파 및 MCP-1 발현을 억제함을 알 수 있다.18 is a diagram showing the experimental results of confirming the TNF-alpha expression inhibitory effect of the Yonggyu water extract, FIG. 19 is a road showing the experimental results confirming the MCP-1 expression inhibitory effect of the Yonggyu water extract, the Yonggyu water extract effectively TNF-alpha and It can be seen that MCP-1 expression is suppressed.
이와 같은 결과로부터, 용규 물추출물이 자궁내막증 및 합병증의 치료, 개선 또는 예방에 효과적임을 확인할 수 있다.From these results, it can be confirmed that the Yonggyu water extract is effective in the treatment, improvement or prevention of endometriosis and complications.
따라서, 용규 추출물은 세포이동 및 부착 관련 효소 발현 억제, 염증 및 통증 관련 효소 발현 억제, 염증성 싸이토카인 발현 억제, 자궁내막증세포 이동 억제, 자궁내막증 세포 부착 및/또는 착상 억제 등에 의해, 자궁내막증 또는 그 합병증의 치료, 개선 및/또는 예방 효과가 있음을 알 수 있다.
Therefore, Yonggyu extract inhibits cell migration and adhesion-related enzyme expression, inflammation and pain-related enzyme expression suppression, inflammatory cytokine expression suppression, endometriosis cell migration suppression, endometriosis cell adhesion and/or implantation suppression, etc., thereby, endometriosis or its complications It can be seen that there is a therapeutic, ameliorating and/or preventing effect of
<제조예 1> 약학 조성물의 제조<Preparation Example 1> Preparation of pharmaceutical composition
실시예 1-1 ~ 실시예 4-1에 기재된 방법 중 어느 하나와 동일한 방법으로 준비한 용규 추출물 300mg, 옥수수 전분 100mg, 유당 100mg, 스테아린산 마그네슘 2mg을 젤라틴 캡슐에 충전하여 캡슐제를 제조하였다.300 mg of Yonggyu extract, 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate prepared in the same manner as in any one of Examples 1-1 to 4-1 were filled into gelatin capsules to prepare capsules.
<제조예 2> 식품 조성물의 제조<Preparation Example 2> Preparation of food composition
실시예 1-1 ~ 실시예 4-1에 기재된 방법 중 어느 하나와 동일한 방법으로 준비한 용규 추출물(4중량 %), 액상과당(0.5중량%), 올리고당(2중량%), 설탕(2중량%), 및 식염(0.5중량%)에 물을 추가하여 잔량을 맞춘 후 균질하게 배합하여 순간 살균을 하여 건강음료를 제조하였다.Yonggyu extract (4% by weight), high fructose (0.5% by weight), oligosaccharide (2% by weight), sugar (2% by weight) prepared in the same manner as in any one of the methods described in Examples 1-1 to 4-1 ), and salt (0.5 wt%) by adding water to adjust the remaining amount, then homogeneously blended and instantaneously sterilized to prepare a health drink.
<110> DAEWOONG PHARMACEUTICAL CO., LTD. University-Industry Cooperation Group of Kyung Hee University <120> Novel use of Extract of Solani Nigri Herba <130> GP15017 <160> 12 <170> KopatentIn 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Sense primer for MMP-2 <400> 1 ccgcagtgac ggaaagatgt 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Antisense primer for MMP-2 <400> 2 cacttgcggt cgtcatcgta 20 <210> 3 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Sense primer for MMP-9 <400> 3 ggacgatgcc tgcaacgt 18 <210> 4 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Antisense primer for MMP-9 <400> 4 caaatacagc tggttcccaa tct 23 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Sense primer for GAPDH <400> 5 ggggctctcc agaacatcat 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Antisense primer for GAPDH <400> 6 caggtcaggt ccaccactga 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Sense primer for COX-2 <400> 7 ccagcacttc acgcatcagt 20 <210> 8 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Antisense primer for COX-2 <400> 8 acgctgtcta gccagagttt cac 23 <210> 9 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Sense primer for TNF-a <400> 9 aagcctgtag cccacctcgt a 21 <210> 10 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Antisense primer for TNF-a <400> 10 ggcaccacta gttggttgtc tttg 24 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Sense primer for MCP-1 <400> 11 gctcatagca gccaccttca 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Antisense primer for MCP-1 <400> 12 ggacacttgc tgctggtgat 20 <110> DAEWOONG PHARMACEUTICAL CO., LTD. University-Industry Cooperation Group of Kyung Hee University <120> Novel use of Extract of Solani Nigri Herba <130> GP15017 <160> 12 <170> KopatentIn 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Sense primer for MMP-2 <400> 1 ccgcagtgac ggaaagatgt 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Antisense primer for MMP-2 <400> 2 cacttgcggt cgtcatcgta 20 <210> 3 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Sense primer for MMP-9 <400> 3 ggacgatgcc tgcaacgt 18 <210> 4 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Antisense primer for MMP-9 <400> 4 caaatacagc tggttcccaa tct 23 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Sense primer for GAPDH <400> 5 ggggctctcc agaacatcat 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Antisense primer for GAPDH <400> 6 caggtcaggt ccaccactga 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Sense primer for COX-2 <400> 7 ccagcacttc acgcatcagt 20 <210> 8 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Antisense primer for COX-2 <400> 8 acgctgtcta gccagagttt cac 23 <210> 9 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Sense primer for TNF-a <400> 9 aagcctgtag cccacctcgt a 21 <210> 10 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Antisense primer for TNF-a <400> 10 ggcaccacta gttggttgtc tttg 24 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Sense primer for MCP-1 <400> 11 gctcatagca gccaccttca 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Antisense primer for MCP-1 <400> 12 ggacacttgc tgctggtgat 20
Claims (13)
상기 용규 추출물은 상기 용규를 용매로 추출한 용매추출물로, 상기 용매는 물과 알코올 혼합물 또는 알코올인 것을 특징으로 하는 자궁내막증 또는 이의 합병증 치료 또는 예방용 약학 조성물.Contains Yonggyu extract as an active ingredient,
The Yonggyu extract is a solvent extract obtained by extracting the Yonggyu with a solvent, and the solvent is a pharmaceutical composition for treating or preventing endometriosis or its complications, characterized in that the solvent is a mixture of water and alcohol or alcohol.
상기 부탄올분획추출물은
(A) 용규를 70부피%에탄올로 추출한 70부피%에탄올추출물을 준비하는 단계;
(B) 상기 (A)단계의 70부피%에탄올추출물에 물을 가하는 단계;
(C) 상기 (B)단계의 물이 가해진 70부피%에탄올추출물을 n-헥산으로 분획추출하는 단계;
(D) 상기 (C)단계의 n-헥산 분획을 제거하고 남은 수층을 디클로로메탄으로 분획추출하는 단계;
(E) 상기 (D)단계의 디클로로메탄 분획을 제거하고 남은 수층을 에틸아세테이트로 분획추출하는 단계; 및
(F) 상기 (E)단계의 에틸아세테이트 분획을 제거하고 남은 수층을 부탄올로 분획추출하는 단계에 의해 수득한 분획추출물인 약학 조성물.8. The method of claim 7,
The butanol fraction extract is
(A) preparing a 70% by volume ethanol extract obtained by extracting Yonggyu with 70% by volume ethanol;
(B) adding water to the 70% by volume ethanol extract of step (A);
(C) fractional extraction of the 70% by volume ethanol extract to which water was added in step (B) with n-hexane;
(D) removing the n-hexane fraction of step (C) and fraction-extracting the remaining aqueous layer with dichloromethane;
(E) removing the dichloromethane fraction of step (D) and fractionally extracting the remaining aqueous layer with ethyl acetate; and
(F) A pharmaceutical composition which is a fractional extract obtained by fractional extraction of the remaining aqueous layer with butanol after removing the ethyl acetate fraction of step (E).
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020150034679A KR102324233B1 (en) | 2015-03-12 | 2015-03-12 | Novel use of Extract of Solani Nigri Herba |
| PCT/KR2016/002427 WO2016144125A2 (en) | 2015-03-12 | 2016-03-11 | Novel use of solanum nigrum l. extract |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020150034679A KR102324233B1 (en) | 2015-03-12 | 2015-03-12 | Novel use of Extract of Solani Nigri Herba |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR20160109725A KR20160109725A (en) | 2016-09-21 |
| KR102324233B1 true KR102324233B1 (en) | 2021-11-11 |
Family
ID=56879669
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020150034679A KR102324233B1 (en) | 2015-03-12 | 2015-03-12 | Novel use of Extract of Solani Nigri Herba |
Country Status (2)
| Country | Link |
|---|---|
| KR (1) | KR102324233B1 (en) |
| WO (1) | WO2016144125A2 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11701379B2 (en) | 2018-03-26 | 2023-07-18 | Rhode Island Hospital | In vitro and in vivo intracellular delivery of siRNA via self-assembled nanopieces |
| CN108324798A (en) * | 2018-05-10 | 2018-07-27 | 李明慧 | A kind of converted products and purposes of S. photeinocarpum |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20060034178A (en) | 2004-10-18 | 2006-04-21 | 한국 한의학 연구원 | A therapeutic agent for aids containing extract of solani nigri herba |
| KR20140099394A (en) * | 2013-02-01 | 2014-08-12 | 한국식품연구원 | Composition for preventing, improving or treating of th1-mediated immune disease or th2-mediated immune disease comprising extracts from solani nigri herba, extracts from dictamni radicis cortex or its combination as an active ingredients |
-
2015
- 2015-03-12 KR KR1020150034679A patent/KR102324233B1/en active IP Right Grant
-
2016
- 2016-03-11 WO PCT/KR2016/002427 patent/WO2016144125A2/en active Application Filing
Non-Patent Citations (3)
| Title |
|---|
| Archives of Gynecology and Obstetrics, 2015년 1월 (온라인 공개일), 292권, 1호, 페이지 21-35 |
| Evidence-Based Complementary and Alternative Medicine, 2012년, 2012권, 논문 번호 859185, 내부페이지 1-10 |
| 고유미 외. 용규가 HeLa cell 증식억제와 apoptosis에 미치는 영향. 대한한방부인과학회지. 2005, Vol. 18(2), pp. 1-11.* |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2016144125A3 (en) | 2016-10-27 |
| KR20160109725A (en) | 2016-09-21 |
| WO2016144125A2 (en) | 2016-09-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR101986229B1 (en) | Composition for Improving, Preventing or Treating Muscular Diseases Comprising Natural Extract | |
| KR102324233B1 (en) | Novel use of Extract of Solani Nigri Herba | |
| KR101956620B1 (en) | Compositions for preventing or treating fibrotic diseases comprising dendropanax morbifera extracts | |
| CN116832081A (en) | Pharmaceutical composition for preventing or treating diabetic complications and angioedema comprising natural mixture extract as active ingredient | |
| KR101621446B1 (en) | Composition for preventing or treating thyroid disorders comprising euphorbia kansui liou ex wang extracts or fraction thereof | |
| KR20110032446A (en) | Composition comprising the extract of astragalus membranaceus bge. ,cinnamomum cassia and phellodendron amurensis for preventing and treating of osteoporesis and bone disease | |
| KR101828167B1 (en) | Composition for anti-obesity comprising the extract of aronia and method for preparing thereof | |
| KR102447045B1 (en) | Composition containing an extract of dendropanax morbifera | |
| KR101357119B1 (en) | A pharmaceutical composition comprising extract of Puerariae Flos for prevention and treatment of endometriosis | |
| KR100456089B1 (en) | The method for preparing purified extract showing anti-cancer activity from wild ginseng and the composition comprising the same | |
| KR20210020818A (en) | Composition for preventing, ameliorating, or treating disease associated with muscle loss, comprising extract of amomum tsaoko | |
| KR102157247B1 (en) | A composition for improving, preventing and treating of pruritus comprising Porphyra yezoensis extract | |
| KR20150060653A (en) | Pharmaceutical compositions for the treatment of cancer metastasis or inhibition of metastasis containing Quassia undulata extracts as active fractions | |
| KR101735242B1 (en) | Composition for preventing and treating male reproductive organ related diseases | |
| KR20070114444A (en) | A composition comprising an extract of meg formulation for the prevention and treatment of diabetes mellitus | |
| KR102475985B1 (en) | Composition for preventing or treating rheumatoid arthritis comprising combination extract containing Rhei Rhizoma, Scutellariae Radix and Coptidis Rhizoma | |
| CN111728985B (en) | A composition containing oviductus Ranae as main ingredient and its application | |
| US20230127213A1 (en) | A topical composition comprising an extract of combined herbs comprising longanae arillus for the treatment or alleviation of skin ulcer and the use thereof | |
| KR102479180B1 (en) | Sanguisorba officinalis Linne extract having a suppressive effect against enzymatic activity of the SARS-CoV-2 3C-like protease and RNA-dependent RNA Polymerase | |
| KR20170036547A (en) | Novel use of alchol extract of Taraxci Herba | |
| KR102119812B1 (en) | Composition for Preventing or Treating Uterine Myoma Comprising Rhus verniciflua Stokes Extract | |
| KR102454513B1 (en) | Composition for preventing and treating arthritis comprising of Artemisia argyi thereof | |
| KR102590557B1 (en) | Composition containing slugs extract as an active ingredient for the prevention, improvement or treatment of male prostate disease | |
| US20230103514A1 (en) | A topical composition comprising an extract of combined herbs comprising longanae arillus for the skin regeneration and the treatment or alleviation of skin wound and the use thereof | |
| KR101215797B1 (en) | A composition comprising a morus extract for preventing and treating liver cirrhosis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| E902 | Notification of reason for refusal | ||
| E701 | Decision to grant or registration of patent right |