WO2021201503A1 - An oral pharmaceutical composition comprising an extract of combined herbs comprising longanae arillus for the treatment or alleviation of arthritis disease and the use thereof. - Google Patents
An oral pharmaceutical composition comprising an extract of combined herbs comprising longanae arillus for the treatment or alleviation of arthritis disease and the use thereof. Download PDFInfo
- Publication number
- WO2021201503A1 WO2021201503A1 PCT/KR2021/003729 KR2021003729W WO2021201503A1 WO 2021201503 A1 WO2021201503 A1 WO 2021201503A1 KR 2021003729 W KR2021003729 W KR 2021003729W WO 2021201503 A1 WO2021201503 A1 WO 2021201503A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- extract
- rhizoma
- tenuissimi
- ligustici
- arthritis
- Prior art date
Links
- 239000000284 extract Substances 0.000 title claims abstract description 87
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title claims abstract description 35
- 206010003246 arthritis Diseases 0.000 title claims abstract description 32
- 239000008203 oral pharmaceutical composition Substances 0.000 title claims abstract description 15
- 235000008216 herbs Nutrition 0.000 title description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 38
- 238000000034 method Methods 0.000 claims description 26
- 238000002360 preparation method Methods 0.000 claims description 26
- 230000036541 health Effects 0.000 claims description 17
- 201000008482 osteoarthritis Diseases 0.000 claims description 14
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims description 12
- 235000013361 beverage Nutrition 0.000 claims description 12
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 12
- 239000004480 active ingredient Substances 0.000 claims description 10
- 235000013305 food Nutrition 0.000 claims description 10
- 235000013376 functional food Nutrition 0.000 claims description 10
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 9
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 9
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 8
- 241000124008 Mammalia Species 0.000 claims description 8
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 7
- 239000000843 powder Substances 0.000 claims description 7
- 239000003826 tablet Substances 0.000 claims description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 6
- 239000002775 capsule Substances 0.000 claims description 5
- 235000015872 dietary supplement Nutrition 0.000 claims description 5
- 239000000839 emulsion Substances 0.000 claims description 5
- 239000002904 solvent Substances 0.000 claims description 5
- 239000000725 suspension Substances 0.000 claims description 5
- 235000011187 glycerol Nutrition 0.000 claims description 4
- 239000008187 granular material Substances 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 4
- 235000013772 propylene glycol Nutrition 0.000 claims description 4
- 229960004063 propylene glycol Drugs 0.000 claims description 4
- 239000006188 syrup Substances 0.000 claims description 4
- 235000020357 syrup Nutrition 0.000 claims description 4
- PUPZLCDOIYMWBV-UHFFFAOYSA-N (+/-)-1,3-Butanediol Chemical compound CC(O)CCO PUPZLCDOIYMWBV-UHFFFAOYSA-N 0.000 claims description 3
- 239000003937 drug carrier Substances 0.000 claims description 3
- 239000006187 pill Substances 0.000 claims description 3
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 claims description 3
- 201000005569 Gout Diseases 0.000 claims description 2
- 206010034464 Periarthritis Diseases 0.000 claims description 2
- 201000002661 Spondylitis Diseases 0.000 claims description 2
- 201000010603 frozen shoulder Diseases 0.000 claims description 2
- 206010039073 rheumatoid arthritis Diseases 0.000 claims description 2
- 241001122767 Theaceae Species 0.000 claims 2
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 claims 1
- 238000012360 testing method Methods 0.000 abstract description 76
- 102000004127 Cytokines Human genes 0.000 abstract description 32
- 108090000695 Cytokines Proteins 0.000 abstract description 32
- 230000002401 inhibitory effect Effects 0.000 abstract description 31
- 239000000203 mixture Substances 0.000 abstract description 31
- 238000000338 in vitro Methods 0.000 abstract description 19
- 230000000694 effects Effects 0.000 abstract description 18
- 206010061218 Inflammation Diseases 0.000 abstract description 17
- 230000004054 inflammatory process Effects 0.000 abstract description 17
- 241000700159 Rattus Species 0.000 abstract description 16
- 230000014509 gene expression Effects 0.000 abstract description 16
- 230000003110 anti-inflammatory effect Effects 0.000 abstract description 15
- 102100031294 Thymic stromal lymphopoietin Human genes 0.000 abstract description 11
- 238000001727 in vivo Methods 0.000 abstract description 11
- 230000003389 potentiating effect Effects 0.000 abstract description 11
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 abstract description 10
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 abstract description 10
- 238000010171 animal model Methods 0.000 abstract description 9
- 238000002474 experimental method Methods 0.000 abstract description 8
- 230000004900 autophagic degradation Effects 0.000 abstract description 7
- 238000013043 cell viability test Methods 0.000 abstract description 7
- 230000002751 inhibitory effect on arthritis Effects 0.000 abstract description 6
- 238000012404 In vitro experiment Methods 0.000 abstract description 5
- 230000002456 anti-arthritic effect Effects 0.000 abstract description 5
- 208000027866 inflammatory disease Diseases 0.000 abstract description 5
- 101000845170 Homo sapiens Thymic stromal lymphopoietin Proteins 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 56
- 239000002158 endotoxin Substances 0.000 description 27
- 229920006008 lipopolysaccharide Polymers 0.000 description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 26
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 21
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 21
- 206010061363 Skeletal injury Diseases 0.000 description 14
- 239000003981 vehicle Substances 0.000 description 13
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- -1 IL-1β Proteins 0.000 description 11
- 241001465754 Metazoa Species 0.000 description 11
- 210000002683 foot Anatomy 0.000 description 11
- 239000013642 negative control Substances 0.000 description 10
- 108010029307 thymic stromal lymphopoietin Proteins 0.000 description 10
- 201000010099 disease Diseases 0.000 description 9
- 239000012153 distilled water Substances 0.000 description 9
- RZEKVGVHFLEQIL-UHFFFAOYSA-N celecoxib Chemical compound C1=CC(C)=CC=C1C1=CC(C(F)(F)F)=NN1C1=CC=C(S(N)(=O)=O)C=C1 RZEKVGVHFLEQIL-UHFFFAOYSA-N 0.000 description 8
- 229960000590 celecoxib Drugs 0.000 description 8
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 8
- 239000013641 positive control Substances 0.000 description 8
- 210000001519 tissue Anatomy 0.000 description 8
- 102000002274 Matrix Metalloproteinases Human genes 0.000 description 7
- 108010000684 Matrix Metalloproteinases Proteins 0.000 description 7
- 208000000114 Pain Threshold Diseases 0.000 description 7
- 238000000605 extraction Methods 0.000 description 7
- 230000002757 inflammatory effect Effects 0.000 description 7
- 238000010603 microCT Methods 0.000 description 7
- 230000037040 pain threshold Effects 0.000 description 7
- 238000011160 research Methods 0.000 description 7
- 239000000796 flavoring agent Substances 0.000 description 6
- 235000013402 health food Nutrition 0.000 description 6
- 238000007489 histopathology method Methods 0.000 description 6
- 230000001965 increasing effect Effects 0.000 description 6
- 230000001939 inductive effect Effects 0.000 description 6
- 230000028709 inflammatory response Effects 0.000 description 6
- 238000002347 injection Methods 0.000 description 6
- 239000007924 injection Substances 0.000 description 6
- 210000000629 knee joint Anatomy 0.000 description 6
- 230000000770 proinflammatory effect Effects 0.000 description 6
- 238000010998 test method Methods 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 239000003795 chemical substances by application Substances 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 238000011049 filling Methods 0.000 description 5
- 235000013355 food flavoring agent Nutrition 0.000 description 5
- 230000005764 inhibitory process Effects 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 239000006186 oral dosage form Substances 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 102000007469 Actins Human genes 0.000 description 4
- 108010085238 Actins Proteins 0.000 description 4
- 102000004072 Beclin-1 Human genes 0.000 description 4
- 108090000524 Beclin-1 Proteins 0.000 description 4
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 4
- 102000007644 Colony-Stimulating Factors Human genes 0.000 description 4
- 108010071942 Colony-Stimulating Factors Proteins 0.000 description 4
- 201000004624 Dermatitis Diseases 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 4
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 4
- 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 4
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 4
- 229960003957 dexamethasone Drugs 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000008101 lactose Substances 0.000 description 4
- 235000019359 magnesium stearate Nutrition 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 210000001616 monocyte Anatomy 0.000 description 4
- 239000008194 pharmaceutical composition Substances 0.000 description 4
- 239000000546 pharmaceutical excipient Substances 0.000 description 4
- 230000002265 prevention Effects 0.000 description 4
- 230000002441 reversible effect Effects 0.000 description 4
- 229930003231 vitamin Natural products 0.000 description 4
- 235000013343 vitamin Nutrition 0.000 description 4
- 239000011782 vitamin Substances 0.000 description 4
- 229940088594 vitamin Drugs 0.000 description 4
- 150000003722 vitamin derivatives Chemical class 0.000 description 4
- 102100040881 60S acidic ribosomal protein P0 Human genes 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 101100387135 Caenorhabditis elegans dex-1 gene Proteins 0.000 description 3
- 229920002261 Corn starch Polymers 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 3
- 108010010803 Gelatin Proteins 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- 101000673456 Homo sapiens 60S acidic ribosomal protein P0 Proteins 0.000 description 3
- 102000015696 Interleukins Human genes 0.000 description 3
- 108010063738 Interleukins Proteins 0.000 description 3
- 108010015302 Matrix metalloproteinase-9 Proteins 0.000 description 3
- 208000002193 Pain Diseases 0.000 description 3
- 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 3
- 229930006000 Sucrose Natural products 0.000 description 3
- 244000269722 Thea sinensis Species 0.000 description 3
- 206010052428 Wound Diseases 0.000 description 3
- 208000027418 Wounds and injury Diseases 0.000 description 3
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 3
- 150000001720 carbohydrates Chemical class 0.000 description 3
- 235000014633 carbohydrates Nutrition 0.000 description 3
- 230000003833 cell viability Effects 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 239000008120 corn starch Substances 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- 238000004108 freeze drying Methods 0.000 description 3
- 235000015203 fruit juice Nutrition 0.000 description 3
- 235000020510 functional beverage Nutrition 0.000 description 3
- 229920000159 gelatin Polymers 0.000 description 3
- 239000008273 gelatin Substances 0.000 description 3
- 235000019322 gelatine Nutrition 0.000 description 3
- 235000011852 gelatine desserts Nutrition 0.000 description 3
- 230000009931 harmful effect Effects 0.000 description 3
- 230000035876 healing Effects 0.000 description 3
- 210000002865 immune cell Anatomy 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 230000002735 inhibitory effect on pain Effects 0.000 description 3
- 229910052500 inorganic mineral Inorganic materials 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 3
- 235000010755 mineral Nutrition 0.000 description 3
- 239000011707 mineral Substances 0.000 description 3
- 230000036407 pain Effects 0.000 description 3
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- 239000000454 talc Substances 0.000 description 3
- 229910052623 talc Inorganic materials 0.000 description 3
- 238000003809 water extraction Methods 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- 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 2
- GHOKWGTUZJEAQD-ZETCQYMHSA-N (D)-(+)-Pantothenic acid Chemical compound OCC(C)(C)[C@@H](O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-ZETCQYMHSA-N 0.000 description 2
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 2
- 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 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- HJXMNVQARNZTEE-UHFFFAOYSA-N Butylphthalide Chemical compound C1=CC=C2C(CCCC)OC(=O)C2=C1 HJXMNVQARNZTEE-UHFFFAOYSA-N 0.000 description 2
- 206010050685 Cytokine storm Diseases 0.000 description 2
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 2
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 2
- 240000001008 Dimocarpus longan Species 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 239000004386 Erythritol Substances 0.000 description 2
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 2
- 235000000235 Euphoria longan Nutrition 0.000 description 2
- 229930091371 Fructose Natural products 0.000 description 2
- 239000005715 Fructose Substances 0.000 description 2
- 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 2
- 102000004269 Granulocyte Colony-Stimulating Factor Human genes 0.000 description 2
- 108010017080 Granulocyte Colony-Stimulating Factor Proteins 0.000 description 2
- 108010002352 Interleukin-1 Proteins 0.000 description 2
- 102000000589 Interleukin-1 Human genes 0.000 description 2
- 108090000176 Interleukin-13 Proteins 0.000 description 2
- 108090000978 Interleukin-4 Proteins 0.000 description 2
- 102000007651 Macrophage Colony-Stimulating Factor Human genes 0.000 description 2
- 108010046938 Macrophage Colony-Stimulating Factor Proteins 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 102100030412 Matrix metalloproteinase-9 Human genes 0.000 description 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
- 241000700157 Rattus norvegicus Species 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 210000001744 T-lymphocyte Anatomy 0.000 description 2
- 244000299461 Theobroma cacao Species 0.000 description 2
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 2
- XLOMVQKBTHCTTD-UHFFFAOYSA-N Zinc monoxide Chemical compound [Zn]=O XLOMVQKBTHCTTD-UHFFFAOYSA-N 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 235000010443 alginic acid Nutrition 0.000 description 2
- 229920000615 alginic acid Polymers 0.000 description 2
- 125000005233 alkylalcohol group Chemical group 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 229910000019 calcium carbonate Inorganic materials 0.000 description 2
- 239000001506 calcium phosphate Substances 0.000 description 2
- 238000010609 cell counting kit-8 assay Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 206010052015 cytokine release syndrome Diseases 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 235000019414 erythritol Nutrition 0.000 description 2
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 2
- 229940009714 erythritol Drugs 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 210000003630 histaminocyte Anatomy 0.000 description 2
- 230000001900 immune effect Effects 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 210000004969 inflammatory cell Anatomy 0.000 description 2
- 229940047122 interleukins Drugs 0.000 description 2
- 210000002414 leg Anatomy 0.000 description 2
- 210000002540 macrophage Anatomy 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 150000004804 polysaccharides Chemical class 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 229960003415 propylparaben Drugs 0.000 description 2
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 description 2
- HELXLJCILKEWJH-NCGAPWICSA-N rebaudioside A Chemical compound O([C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HELXLJCILKEWJH-NCGAPWICSA-N 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000010992 reflux Methods 0.000 description 2
- 238000009877 rendering Methods 0.000 description 2
- 238000003757 reverse transcription PCR Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- AGDSCTQQXMDDCV-UHFFFAOYSA-M sodium;2-iodoacetate Chemical compound [Na+].[O-]C(=O)CI AGDSCTQQXMDDCV-UHFFFAOYSA-M 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000000600 sorbitol Substances 0.000 description 2
- 235000010356 sorbitol Nutrition 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 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 2
- 235000000346 sugar Nutrition 0.000 description 2
- 239000010414 supernatant solution Substances 0.000 description 2
- 239000000829 suppository Substances 0.000 description 2
- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 description 2
- 210000000689 upper leg Anatomy 0.000 description 2
- 239000000080 wetting agent Substances 0.000 description 2
- 230000029663 wound healing Effects 0.000 description 2
- 230000037314 wound repair Effects 0.000 description 2
- 239000000811 xylitol Substances 0.000 description 2
- 235000010447 xylitol Nutrition 0.000 description 2
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 2
- 229960002675 xylitol Drugs 0.000 description 2
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 241000208173 Apiaceae Species 0.000 description 1
- 108010011485 Aspartame Proteins 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- PTHCMJGKKRQCBF-UHFFFAOYSA-N Cellulose, microcrystalline Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC)C(CO)O1 PTHCMJGKKRQCBF-UHFFFAOYSA-N 0.000 description 1
- 102000019034 Chemokines Human genes 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- GHOKWGTUZJEAQD-UHFFFAOYSA-N Chick antidermatitis factor Natural products OCC(C)(C)C(O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-UHFFFAOYSA-N 0.000 description 1
- 235000005979 Citrus limon Nutrition 0.000 description 1
- 244000131522 Citrus pyriformis Species 0.000 description 1
- UXDIXFDKSPCUIX-AXFHLTTASA-N Cnidilide Chemical compound C1=CCC[C@@H]2[C@H](CCCC)OC(=O)[C@@H]21 UXDIXFDKSPCUIX-AXFHLTTASA-N 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 206010012438 Dermatitis atopic Diseases 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 208000008960 Diabetic foot Diseases 0.000 description 1
- 235000019739 Dicalciumphosphate Nutrition 0.000 description 1
- 235000000525 Dimocarpus longan Nutrition 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
- 206010015866 Extravasation Diseases 0.000 description 1
- 239000001512 FEMA 4601 Substances 0.000 description 1
- 101100175482 Glycine max CG-3 gene Proteins 0.000 description 1
- 239000004378 Glycyrrhizin Substances 0.000 description 1
- 101001008922 Homo sapiens Kallikrein-11 Proteins 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 102000008070 Interferon-gamma Human genes 0.000 description 1
- 108010074328 Interferon-gamma Proteins 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 108010002616 Interleukin-5 Proteins 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 208000012659 Joint disease Diseases 0.000 description 1
- 102100027612 Kallikrein-11 Human genes 0.000 description 1
- 240000007472 Leucaena leucocephala Species 0.000 description 1
- 235000010643 Leucaena leucocephala Nutrition 0.000 description 1
- 241000244359 Ligusticum jeholense Species 0.000 description 1
- 241000244365 Ligusticum sinense Species 0.000 description 1
- 241000143634 Ligusticum tenuissimum Species 0.000 description 1
- 102000008072 Lymphokines Human genes 0.000 description 1
- 108010074338 Lymphokines Proteins 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 102000001776 Matrix metalloproteinase-9 Human genes 0.000 description 1
- 102000005741 Metalloproteases Human genes 0.000 description 1
- 108010006035 Metalloproteases Proteins 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 241000906034 Orthops Species 0.000 description 1
- 229920002230 Pectic acid Polymers 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- 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 1
- 206010034972 Photosensitivity reaction Diseases 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 241001080798 Polygala tenuifolia Species 0.000 description 1
- 241000208977 Polygalaceae Species 0.000 description 1
- 244000018633 Prunus armeniaca Species 0.000 description 1
- 235000009827 Prunus armeniaca Nutrition 0.000 description 1
- 239000012979 RPMI medium Substances 0.000 description 1
- HELXLJCILKEWJH-SEAGSNCFSA-N Rebaudioside A Natural products O=C(O[C@H]1[C@@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1)[C@@]1(C)[C@@H]2[C@](C)([C@H]3[C@@]4(CC(=C)[C@@](O[C@H]5[C@H](O[C@H]6[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O6)[C@@H](O[C@H]6[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O6)[C@H](O)[C@@H](CO)O5)(C4)CC3)CC2)CCC1 HELXLJCILKEWJH-SEAGSNCFSA-N 0.000 description 1
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 1
- 241001093760 Sapindaceae Species 0.000 description 1
- 206010040943 Skin Ulcer Diseases 0.000 description 1
- 244000228451 Stevia rebaudiana 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
- 102100040247 Tumor necrosis factor Human genes 0.000 description 1
- 206010054094 Tumour necrosis Diseases 0.000 description 1
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 1
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 229930003451 Vitamin B1 Natural products 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 208000038016 acute inflammation Diseases 0.000 description 1
- 230000006022 acute inflammation Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 201000009961 allergic asthma Diseases 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000003356 anti-rheumatic effect Effects 0.000 description 1
- 230000036528 appetite Effects 0.000 description 1
- 235000019789 appetite Nutrition 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000003125 aqueous solvent Substances 0.000 description 1
- 230000002917 arthritic effect Effects 0.000 description 1
- 239000000605 aspartame Substances 0.000 description 1
- 235000010357 aspartame Nutrition 0.000 description 1
- IAOZJIPTCAWIRG-QWRGUYRKSA-N aspartame Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(=O)OC)CC1=CC=CC=C1 IAOZJIPTCAWIRG-QWRGUYRKSA-N 0.000 description 1
- 229960003438 aspartame Drugs 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 239000012298 atmosphere Substances 0.000 description 1
- 201000008937 atopic dermatitis Diseases 0.000 description 1
- 230000004642 autophagic pathway Effects 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000008512 biological response Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 235000015895 biscuits Nutrition 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 235000008429 bread Nutrition 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000004067 bulking agent Substances 0.000 description 1
- 235000014121 butter Nutrition 0.000 description 1
- 238000010805 cDNA synthesis kit Methods 0.000 description 1
- 235000001046 cacaotero Nutrition 0.000 description 1
- 235000001465 calcium Nutrition 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 239000000378 calcium silicate Substances 0.000 description 1
- 229910052918 calcium silicate Inorganic materials 0.000 description 1
- 235000012241 calcium silicate Nutrition 0.000 description 1
- OYACROKNLOSFPA-UHFFFAOYSA-N calcium;dioxido(oxo)silane Chemical compound [Ca+2].[O-][Si]([O-])=O OYACROKNLOSFPA-UHFFFAOYSA-N 0.000 description 1
- 238000010000 carbonizing Methods 0.000 description 1
- 230000003293 cardioprotective effect Effects 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 210000000845 cartilage Anatomy 0.000 description 1
- 230000005754 cellular signaling Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 235000013351 cheese Nutrition 0.000 description 1
- 229940112822 chewing gum Drugs 0.000 description 1
- 235000015218 chewing gum Nutrition 0.000 description 1
- 235000019219 chocolate Nutrition 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 230000006020 chronic inflammation Effects 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- UXDIXFDKSPCUIX-UHFFFAOYSA-N condilide Natural products C1=CCCC2C(CCCC)OC(=O)C21 UXDIXFDKSPCUIX-UHFFFAOYSA-N 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000012050 conventional carrier Substances 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 235000012495 crackers Nutrition 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000001934 delay Effects 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- NEFBYIFKOOEVPA-UHFFFAOYSA-K dicalcium phosphate Chemical compound [Ca+2].[Ca+2].[O-]P([O-])([O-])=O NEFBYIFKOOEVPA-UHFFFAOYSA-K 0.000 description 1
- 229910000390 dicalcium phosphate Inorganic materials 0.000 description 1
- 229940038472 dicalcium phosphate Drugs 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000002905 effect on arthritis Effects 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- HELXLJCILKEWJH-UHFFFAOYSA-N entered according to Sigma 01432 Natural products C1CC2C3(C)CCCC(C)(C(=O)OC4C(C(O)C(O)C(CO)O4)O)C3CCC2(C2)CC(=C)C21OC(C1OC2C(C(O)C(O)C(CO)O2)O)OC(CO)C(O)C1OC1OC(CO)C(O)C(O)C1O HELXLJCILKEWJH-UHFFFAOYSA-N 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
- 239000003172 expectorant agent Substances 0.000 description 1
- 230000003419 expectorant effect Effects 0.000 description 1
- 230000036251 extravasation Effects 0.000 description 1
- 210000003414 extremity Anatomy 0.000 description 1
- 239000011790 ferrous sulphate Substances 0.000 description 1
- 235000003891 ferrous sulphate Nutrition 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 239000005417 food ingredient Substances 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 239000007903 gelatin capsule Substances 0.000 description 1
- 230000008570 general process Effects 0.000 description 1
- LPLVUJXQOOQHMX-UHFFFAOYSA-N glycyrrhetinic acid glycoside Natural products C1CC(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2=O)C(O)=O)C)(C)CC2)(C)C2C(C)(C)C1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O LPLVUJXQOOQHMX-UHFFFAOYSA-N 0.000 description 1
- 229960004949 glycyrrhizic acid Drugs 0.000 description 1
- UYRUBYNTXSDKQT-UHFFFAOYSA-N glycyrrhizic acid Natural products CC1(C)C(CCC2(C)C1CCC3(C)C2C(=O)C=C4C5CC(C)(CCC5(C)CCC34C)C(=O)O)OC6OC(C(O)C(O)C6OC7OC(O)C(O)C(O)C7C(=O)O)C(=O)O UYRUBYNTXSDKQT-UHFFFAOYSA-N 0.000 description 1
- 235000019410 glycyrrhizin Nutrition 0.000 description 1
- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 210000000548 hind-foot Anatomy 0.000 description 1
- 238000007602 hot air drying Methods 0.000 description 1
- 235000015243 ice cream Nutrition 0.000 description 1
- 208000026278 immune system disease Diseases 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000002718 inhibitory effect on inflammation Effects 0.000 description 1
- 230000015788 innate immune response Effects 0.000 description 1
- 210000005007 innate immune system Anatomy 0.000 description 1
- 229960003130 interferon gamma Drugs 0.000 description 1
- 229940047124 interferons Drugs 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 239000002085 irritant Substances 0.000 description 1
- 231100000021 irritant Toxicity 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- 229940041476 lactose 100 mg Drugs 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
- 210000000265 leukocyte Anatomy 0.000 description 1
- 210000003041 ligament Anatomy 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 229940057995 liquid paraffin Drugs 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 229960003511 macrogol Drugs 0.000 description 1
- ZLNQQNXFFQJAID-UHFFFAOYSA-L magnesium carbonate Chemical compound [Mg+2].[O-]C([O-])=O ZLNQQNXFFQJAID-UHFFFAOYSA-L 0.000 description 1
- 239000001095 magnesium carbonate Substances 0.000 description 1
- 229910000021 magnesium carbonate Inorganic materials 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 239000000845 maltitol Substances 0.000 description 1
- 235000010449 maltitol Nutrition 0.000 description 1
- VQHSOMBJVWLPSR-WUJBLJFYSA-N maltitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-WUJBLJFYSA-N 0.000 description 1
- 229940035436 maltitol Drugs 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 235000013622 meat product Nutrition 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 description 1
- 239000004292 methyl p-hydroxybenzoate Substances 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- 229960002216 methylparaben Drugs 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 239000002480 mineral oil Substances 0.000 description 1
- 235000010446 mineral oil Nutrition 0.000 description 1
- 230000002297 mitogenic effect Effects 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 230000003040 nociceptive effect Effects 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 229940100688 oral solution Drugs 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 229940055726 pantothenic acid Drugs 0.000 description 1
- 235000019161 pantothenic acid Nutrition 0.000 description 1
- 239000011713 pantothenic acid Substances 0.000 description 1
- 239000006201 parenteral dosage form Substances 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- LCLHHZYHLXDRQG-ZNKJPWOQSA-N pectic acid Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)O[C@H](C(O)=O)[C@@H]1OC1[C@H](O)[C@@H](O)[C@@H](OC2[C@@H]([C@@H](O)[C@@H](O)[C@H](O2)C(O)=O)O)[C@@H](C(O)=O)O1 LCLHHZYHLXDRQG-ZNKJPWOQSA-N 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 235000013550 pizza Nutrition 0.000 description 1
- 229920002492 poly(sulfone) Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000010318 polygalacturonic acid Substances 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 230000031339 positive regulation of inflammatory response Effects 0.000 description 1
- 239000001508 potassium citrate Substances 0.000 description 1
- 229960002635 potassium citrate Drugs 0.000 description 1
- QEEAPRPFLLJWCF-UHFFFAOYSA-K potassium citrate (anhydrous) Chemical compound [K+].[K+].[K+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O QEEAPRPFLLJWCF-UHFFFAOYSA-K 0.000 description 1
- 235000011082 potassium citrates Nutrition 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 230000008741 proinflammatory signaling process Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 description 1
- 239000004405 propyl p-hydroxybenzoate Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- RADKZDMFGJYCBB-UHFFFAOYSA-N pyridoxal hydrochloride Natural products CC1=NC=C(CO)C(C=O)=C1O RADKZDMFGJYCBB-UHFFFAOYSA-N 0.000 description 1
- 238000011552 rat model Methods 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 235000019203 rebaudioside A Nutrition 0.000 description 1
- 230000011514 reflex Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 229960000342 retinol acetate Drugs 0.000 description 1
- 235000019173 retinyl acetate Nutrition 0.000 description 1
- 239000011770 retinyl acetate Substances 0.000 description 1
- QGNJRVVDBSJHIZ-QHLGVNSISA-N retinyl acetate Chemical compound CC(=O)OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C QGNJRVVDBSJHIZ-QHLGVNSISA-N 0.000 description 1
- CVHZOJJKTDOEJC-UHFFFAOYSA-N saccharin Chemical compound C1=CC=C2C(=O)NS(=O)(=O)C2=C1 CVHZOJJKTDOEJC-UHFFFAOYSA-N 0.000 description 1
- 235000019204 saccharin Nutrition 0.000 description 1
- 229940081974 saccharin Drugs 0.000 description 1
- 239000000901 saccharin and its Na,K and Ca salt Substances 0.000 description 1
- OARRHUQTFTUEOS-UHFFFAOYSA-N safranin Chemical compound [Cl-].C=12C=C(N)C(C)=CC2=NC2=CC(C)=C(N)C=C2[N+]=1C1=CC=CC=C1 OARRHUQTFTUEOS-UHFFFAOYSA-N 0.000 description 1
- 235000013580 sausages Nutrition 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 235000011888 snacks Nutrition 0.000 description 1
- HRZFUMHJMZEROT-UHFFFAOYSA-L sodium disulfite Chemical compound [Na+].[Na+].[O-]S(=O)S([O-])(=O)=O HRZFUMHJMZEROT-UHFFFAOYSA-L 0.000 description 1
- 229940001584 sodium metabisulfite Drugs 0.000 description 1
- 235000010262 sodium metabisulphite Nutrition 0.000 description 1
- 229960002920 sorbitol Drugs 0.000 description 1
- 235000014347 soups Nutrition 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 210000002536 stromal cell Anatomy 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 150000005846 sugar alcohols Chemical class 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
- 230000008961 swelling Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 229960003080 taurine Drugs 0.000 description 1
- 235000013616 tea Nutrition 0.000 description 1
- 239000000892 thaumatin Substances 0.000 description 1
- 235000010436 thaumatin Nutrition 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
- 230000000451 tissue damage Effects 0.000 description 1
- 231100000827 tissue damage Toxicity 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 238000007492 two-way ANOVA Methods 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 239000011720 vitamin B Substances 0.000 description 1
- 235000010374 vitamin B1 Nutrition 0.000 description 1
- 239000011691 vitamin B1 Substances 0.000 description 1
- 239000011716 vitamin B2 Substances 0.000 description 1
- 235000019158 vitamin B6 Nutrition 0.000 description 1
- 239000011726 vitamin B6 Substances 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 229940011671 vitamin b6 Drugs 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000011787 zinc oxide Substances 0.000 description 1
Classifications
-
- 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
- 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)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
- A61K36/232—Angelica
-
- 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)
- A61K36/69—Polygalaceae (Milkwort family)
-
- 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)
- A61K36/77—Sapindaceae (Soapberry family), e.g. lychee or soapberry
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0087—Galenical forms not covered by A61K9/02 - A61K9/7023
- A61K9/0095—Drinks; Beverages; Syrups; Compositions for reconstitution thereof, e.g. powders or tablets to be dispersed in a glass of water; Veterinary drenches
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
Definitions
- the present invention is related to an oral pharmaceutical composition comprising an extract of combined herbs comprising Longanae arillus for the treatment or prevention of arthritis diseases and the use thereof.
- an inflammatory response is a normal response of human body associated with an edema, a pain etc in case that a tissue or a cell received any invasion causing some organic change in the tissue or cell.
- various kinds of cytokines have been found to be involved in the inflammatory disease.
- cytokines such as TNF- ⁇ , IL-1 ⁇ , IL-6 etc and MMPs (matrix metalloproteinases, such as PDGF, VEGF, and IGF
- MMPs matrix metalloproteinases
- MMPs matrix metalloproteinases
- TIMPs tissue inhibitors of metalloproteinases
- Ligustici Tenuissimi Rhizoma, a rhizoma or root of Ligusticum tenuissimum Kitagawa, Ligusticum sinense Oliv, Ligusticum jeholense Nakai et Kitagawa or the same species belonged to Umbelliferae has been reported to contain a cnidilide, 3-butyl phthalide etc and to show anti-bacterial effect etc (Chung B. S et al, Dohaehyangyakdaesajeon, youngrimsa, 2 nd Ed. P428-429, 1998).
- Polygalae radix a root of Polygala tenuifolia Willd., or the same species belonged to Polygalaceae has been reported to contain various sanponis and to show expectorant activity, anti-bacterial effect etc (Chung B. S et al, Dohaehyangyakdaesajeon, youngrimsa, 2 nd Ed. P798-799, 1998).
- the technical solution to solve the problem of the background art is for the development of novel herb formulation for treating and preventing inflammation disease or arthritis diseases.
- an oral pharmaceutical composition comprising a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix as an active ingredient to treat and alleviate arthritis diseases.
- extract comprises the extract which can be extracted with at least one solvent selected from water, C1-C4 lower alkyl alcohol such as methanol, ethanol, propanol, butanol, etc, acetone, ethyl acetate, chloroform, hexane, butyleneglycol, propyleneglycol or glycerin, preferably, water, methanol, ethanol, more preferably, water or 10-90 %(v/v) ethanol in water, most preferably, water or 20-80 %(v/v) ethanol in water.
- solvent selected from water, C1-C4 lower alkyl alcohol such as methanol, ethanol, propanol, butanol, etc, acetone, ethyl acetate, chloroform, hexane, butyleneglycol, propyleneglycol or glycerin, preferably, water, methanol, ethanol, more preferably, water or 10-90 %(v/v) ethanol
- arthritic diseases comprises the disease selected from rigid spondylitis, degenerative arthritis, osteoarthritis, rheumatoid arthritis, gout or frozen shoulder etc.
- anti-Inflammation means all the mechanism to inhibit various inflammation.
- anti-arthritic means all the mechanism to inhibit various rheumatism.
- Inflammation is part of the complex biological response of body tissues to harmful stimuli, such as pathogens, damaged cells, or irritants, and the nonspecific immune response such as heat, pain, redness, swelling, etc is called as “inflammatory response”.
- Inflammation can be classified as (a) acute inflammation, the initial response of the body to harmful stimuli, is achieved by the increased movement of plasma and leukocytes (especially granulocytes) from the blood into the injured tissues and then a series of biochemical events propagates and matures the inflammatory response, involving the local vascular system, the immune system, and various cells within the injured tissue and (b)Prolonged inflammation, known as chronic inflammation, leading to a progressive shift in the type of cells present at the site of inflammation, such as mononuclear cells, and being characterized by simultaneous destruction and healing of the tissue from the inflammatory process.
- the macrophage in damaged cell excrete various cytokines, which activates T lymphocyte and mast cell, a lymphocyte, releases various histamines, which initiate internal barrier response, resulting in inducing inflammation of the inflected cells. Accordingly, the expressed level of cell cytokines may be used as an indicator of the activation of inflammatory response (the other aspects, anti-inflammatory activity).
- the “anti-inflammatory activity” disclosed herein denotes the inhibitory activity against various skin inflammation.
- Cytokines means all the immunological substances including chemokines, interferons, interleukins, lymphokines, and tumour necrosis factors produced by a broad range of cells, including immune cells like macrophages, B lymphocytes, T lymphocytes and mast cells, as well as endothelial cells, fibroblasts, and various stromal cells, which are released through immunological progress caused by the infiltration of various pathogen such as virus etc.
- cytokine storm is a physiological reaction in which the innate immune system causes an uncontrolled and excessive release of pro-inflammatory signaling molecules called cytokines and it exacerbates the inflammation resulting from the extremely abundant homing of immune cells to the inflected area, causes to blood extravasation through the loosening of blood vessel and severely to death.
- the term “the inhibitory activity of cytokine expression” disclosed herein can be interpreted as a prevention, treatment or improvement of cytokine storm.
- cytokine comprises various cytokine involved in dermatitis, such as atopic dermatitis, specifically, the cytokine selected from group of TLSP (thymic stromal lymphopoietin), colony stimulating factor (CSF) such as GM-CSF (granulocyte-macrophage colony stimulating factor), M-CSF (macrophage colony stimulating factor), G-CSF (granulocyte colony stimulating factor) and the like, interleukins such as interleukin-1 (IL-1), IL-4, IL-10, IL-12, IL-13, IL-31, IL-33 and the like, tumor necrosis factor alpha (TNF- ⁇ ), interferon gamma (IFN ⁇ ) etc,
- TLSP thymic stromal lymphopoietin
- CSF colony stimulating factor
- GM-CSF granulocyte-macrophage colony stimulating factor
- M-CSF macrophage colony stimulating factor
- An inventive extract may be prepared in accordance with the following preferred embodiment.
- combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix can be prepared by the procedure comprising the steps; of slicing and washing Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix” to use as a basic extraction material at 1 st step; mixing together thoroughly with the mixed ratio based on the dried weight of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix (w/w) ranging from 0.01 -100: 0.01 -100: 0.01 -100 weight part (w/w), preferably, 0.1-50: 0.1-50: 0.1-50 weight part (w/w), more preferably, 0.1-10: 0.1-10: 0.1-10 weight part (w/w), more and more preferably, 1-5: 1-5: 1-5 weight part (w/w), most preferably, 1-3: 1-3 : 1-3 weight
- It is another object of the present invention to provide an oral pharmaceutical composition comprising a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix prepared by the above-described process, as an active ingredient to treat and alleviate arthritis diseases.
- a method of treating or alleviating arthritis diseases in a mammal comprising orally administering to said mammal an effective amount of the combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix and pharmaceutically acceptable carrier thereof.
- inventive composition is potent by accomplishing in vitro experiments such as the inhibitory test on the expression of cytokines involved in inflammation (RPLPO, TSLP, GM-CSF and IL-1beta) (Experimental Example 1); Cell viability test on HT-29 and THP-1 cell (in vitro Experimental Example 2); Anti-inflammatory activity in THP-1 cell (in vitro, Experimental Example 3); inhibitory effect on autophagy activity (in vitro, Experimental Example 4.) as well as in vivo experiments such as inhibitory effect on arthritis using by arthritis-induced rat animal model (in vivo, Experimental Example 5), therefore, it is confirmed that inventive combined extract is very useful in the alleviation or treatment of inflammatory disease and arthritis disease as a form of oral pharmaceutical composition.
- inventive combined extract is very useful in the alleviation or treatment of inflammatory disease and arthritis disease as a form of oral pharmaceutical composition.
- the pharmaceutical composition for treating purposed diseases could contain about 0.01 to 99 w/w% the above herb extract of the present invention based on the total weight of the composition.
- the amount and each component of the above-mentioned composition can be varied with the patient’s condition, development of patient’s disease, the sort of disease etc.
- the inventive composition may additionally comprise conventional carrier, adjuvants or diluents in accordance with a using method.
- the herb composition according to the present invention can be formulated in oral dosage form such as powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol and the like; topical preparation; or injection solution.
- the herb composition according to the present invention can be provided as a pharmaceutical composition containing pharmaceutically acceptable carriers, adjuvants or diluents, e.g., lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starches, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, magnesium stearate and mineral oil.
- pharmaceutically acceptable carriers, adjuvants or diluents e.g., lactose, dextrose, sucrose,
- the formulations may additionally include excipients such as fillers, bulking agents, binders, wetting agents, disintegrating agents, surfactants, diluents and the like.
- the solid oral dosage form comprises tablet, pill, powder, granule, capsule and the like and the solid oral dosage form is prepared by adding at least one excipient such as starch, calcium carbonate, sucrose, lactose or gelatin and the like to the herb extract.
- Lubricant such as magnesium stearate or talc may be used.
- the aqueous oral dosage form comprises suspension, oral solution, emulsion, syrup and the aqueous oral dosage form may comprise several excipients such as wetting agents, sweetener flavoring agents, preservatives, as well as water, liquid paraffin.
- the parenteral dosage form comprises sterilized aqueous solution, non-aqueous solvent, suspension, emulsion, lyophilized preparation, suppository, and the like.
- Suitable examples of the carriers include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable ester such as ethyl oleate.
- Base for suppository may include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatine etc., but are not limited to them.
- the desirable dose of the inventive composition varies depending on the condition and the weight of the subject, severity, drug form, route and period of administration, and may be chosen by those skilled in the art. However, in order to obtain desirable effects, it is generally recommended to administer at the amount ranging 0.01mg/kg to10g/kg, preferably, 1mg/kg to 1g/kg by weight/day of the inventive composition of the present invention.
- the dose may be administered in a single or multiple doses per day.
- composition of present invention can be administered to a subject animal such as mammals (rat, mouse, domestic animals or human) via various routes. All modes of administration are contemplated, for example, administration can be made orally, rectally or by intravenous injection.
- a health functional food comprising a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix, as an active ingredient to prevent or improve arthritis diseases.
- a health functional food defined herein comprises the functional food having enhanced functionality such as physical functionality or physiological functionality by adding the extract of the present invention to conventional food to prevent or improve the purposed diseases in human or mammal and stipulated by the Law for Health Functional Foods 6727 in Republic of Korea.
- the health functional food composition for preventing and improving purposed diseases could contain about 0.01 to 95 w/w%, preferably 1 to 80 w/w% of the above herb composition of present invention based on the total weight of the composition.
- inventive extract of the present invention also can be used as a main component or additive and aiding agent in the preparation of various functional health food and health supplement food for the prevention or improvement of arthritis diseases.
- the inventive health functional food may be prepared and processed by the form of pharmaceutically acceptable dosage form such as powder, granule, tablet, capsule, pills, suspension, emulsion, syrup and the like; or the functional health food form such as tea bag, leached tea, health beverage type and the like.
- pharmaceutically acceptable dosage form such as powder, granule, tablet, capsule, pills, suspension, emulsion, syrup and the like
- functional health food form such as tea bag, leached tea, health beverage type and the like.
- the above health supplement food comprises about 30 to 99 (w/w%), preferably 50 to 99 (w/w%), more preferably 70 to 99 (w/w%) of the inventive extract of present invention based on the total weight of the composition.
- the health functional beverage composition can comprise other component such as flavoring agent or natural carbohydrate without limits like that typical beverage composition.
- natural carbohydrate comprise monosaccharide such as glucose, fructose etc; disaccharide such as maltose, sucrose etc; and polysaccharide, for example, sugar such as dextrin, cyclodextrin, and sugar alcohol such as xylitol, sorbitol, erythritol.
- Natural flavoring agent thaumatin, stevia extract (rebaudioside A, glycyrrhizin, etc)
- synthetic flavoring agent sacharin, aspartame, etc
- the amount of natural carbohydrate generally ranges from about 1 to 20g, preferably about 5 to 12g per 100ml of the present composition.
- the combined herb extract of the present invention When used as a food additive of the health food, the combined herb extract may be added intact or used with other food ingredient according to general process.
- the food comprises meat products, sausage, bread, chocolate, candy, snack, cracker, biscuit, pizza, ramen, noodle products, chewing gum, dairy products such as ice cream, soup, beverage, tea, drinks, alcohol drink, vitamin complex etc, but not intended herein to limit thereto, for preventing or improving of purposed disease.
- the other components than aforementioned composition are various nutrients, a vitamin, a mineral or an electrolyte, synthetic flavoring agent, a coloring agent and improving agent in case of cheese, chocolate et al., pectic acid and the salt thereof, alginic acid and the salt thereof, organic acid, protective colloidal adhesive, pH controlling agent, stabilizer, a preservative, glycerin, alcohol, carbonizing agent used in carbonate beverage et al.
- the other component than aforementioned ones may be fruit juice for preparing natural fruit juice, fruit juice beverage and vegetable beverage, wherein the component can be used independently or in combination.
- the ratio of the components is not so important but is generally range from about 0 to 20 w/w % per 100 w/w % present composition.
- above described extract can be added to food or beverage for prevention and improvement of purposed disorder.
- the amount of above described extract in food or beverage as a functional health food or health supplement food may generally range from about 0.01 to 15 w/w % of total weight of food for functional health food composition.
- the extract of the present invention may be added 0.02 to 5g, preferably 0.3 to 1g per 100ml in health beverage composition.
- inventive combined composition is potent by accomplishing in vitro experiments such as the inhibitory test on the expression of cytokines involved in skin inflammation (RPLPO, TSLP, GM-CSF and IL-1beta) (Experimental Example 1); Cell viability test on HT-29 and THP-1 cell (in vitro Experimental Example 2); Anti-inflammatory activity in THP-1 cell (in vitro, Experimental Example 3); inhibitory effect on autophagy activity (in vitro, Experimental Example 4.) as well as in vivo experiments such as inhibitory effect on arthritis using by arthritis-induced rat animal model (in vivo, Experimental Example 5), therefore, it is confirmed that inventive combined extract is very useful in the alleviation or treatment of inflammatory disease and arthritis disease as a form of oral pharmaceutical composition.
- inventive combined extract is very useful in the alleviation or treatment of inflammatory disease and arthritis disease as a form of oral pharmaceutical composition.
- Example 2-6 The preparation of inventive combined extract (2)-(6)
- Example 2 Excepting adopting different combined ratio as well as different solvents disclosed in Example 1, all the procedure was identical with those in Example 1 to obtain various inventive combined extract of Longanae Arillus (LA) , Ligustici Tenuissimi Rhizoma (LT) and Polygalae radix (PR) i.e., inventive combined extract (2) to inventive combined extract (6) of the present invention, which are used as a test samples in following experiment.
- LA Longanae Arillus
- LT Ligustici Tenuissimi Rhizoma
- PR Polygalae radix
- Example LA* PR* LT* solvent* name Extract weight Final yield
- Example 2 10 5 50 10% EtOH WIN-1002X 16.6g 25.6%
- Example 3 20 50 5 Water WIN-1003X 24.7g 32.9%
- Example 4 10 80 20 70% BuOH WIN-1004X 32.3g 29.4%
- Example 5 50 20 50% EtOH WIN-1005X 21.5g 28.7%
- Example 6 30 10 2 hexane WIN-1006X 12.6g 30.1% *: Longanae Arillus (LA) , Ligustici Tenuissimi Rhizoma (LT), Polygalae radix (PR)
- HaCaT cell human epithelial keratinocyte cell, 300493, CLS
- DMEM medium containing 10% Fetal bovine serum, 100units/ml of penicillin, 100 ⁇ g/ml of streptomycin (D6429, Sigma-Aldrich Co. Ltd) and was incubated in the incubator (HERA cell 150i, Thermo Fisher Scientific Co. Ltd.) maintaining optimum humidity (85-95%) and 5% CO 2 atmosphere.
- TNF alpha RC214-12, Biobasic Co. Ltd
- Dexamethasone 200nM, positive control, “DEX”, D4902, Sigma-Aldrich Co. Ltd.
- DIW distilled water
- RNA FATRR-001, Favorgen
- RRO36A cDNA synthesis kit
- RTPM Enzynomics
- test sample group treated with the inventive extract sharply inhibited the expressed level of various cytokine involved in inflammation comparing with negative control group treated with distilled water (DIW) and it has been confirmed that the inhibitory activity of the test sample on the expression of various cytokine involved in inflammation is equivalent to that of positive control group treated with dexamethasone (DEX).
- DIW negative control group treated with distilled water
- HT-29 cell human colon epithelial cell, Korean Cell Line Bank, Korean Cell Line Research Foundation, 101, Daehak-ro, Jongno-gu, Seoul, 03080, Korea
- DMEM medium containing 10% Fetal bovine serum and 1% penicillin-streptomycin solution
- THP-1 cell human monocyte cell, Korean Cell Line Bank, Korean Cell Line Research Foundation, 101, Daehak-ro, Jongno-gu, Seoul, 03080, Korea
- RPMI medium containing 10% Fetal bovine serum and 1% penicillin-streptomycin solution to incubate.
- test samples were added to HT-29 cell and THP-1 cell. 24 hours after the incubation, CCK-8 (cell counting kit-8, Dojindo Molecular Technologies, Inc.) was added thereto and the absorbance (optical density) was determined at 450nm to determine cell viability.
- CCK-8 cell counting kit-8, Dojindo Molecular Technologies, Inc.
- test sample group treated with the inventive extract (25-1000 ⁇ g/mL) in HT-29 cell showed similar to that of negative control group treated with only medium and that in THP-1 cell showed 90.5% and 26.5% at the concentration of 500 and 1000 ⁇ g/mL, respectively, which showed some different with that of negative control group and less than 500 ⁇ g/mL of test sample was applied to following test.
- cell viability test result WIN1001X ⁇ g/mL HT-29 cell THP-1 cell 0 100 % 100 % 25 100.5 ⁇ 0.6 % 102.2 ⁇ 1.1 % 50 103.3 ⁇ 3.2 % 103.7 ⁇ 1.5 % 100 105.0 ⁇ 1.4 % 104.4 ⁇ 1.6 % 250 109.2 ⁇ 3.1 % 101.1 ⁇ 0.6 % 500 105.9 ⁇ 3.3 % 90.5 ⁇ 0.8 % 1000 90.3 ⁇ 0.1 % 26.5 ⁇ 5.8 %
- LPS lipopolysaccharide
- the level of IL-1beta, an inflammatory cytokine, in the collected cell supernatant solution was measured.
- test samples were treated in THP-1 cells for four hours and LPS was treated therewith to confirm the anti-inflammatory effect of the test sample.
- IL-1beta a pro-inflammatory cytokine
- ELISA reader IL-1beta/IL-1F2 Duo set ELISA, R&D Systems
- test sample prepared in Examples sharply reduced the level of IL-1beta which had been increased with the dose-dependently increased LPS.
- the inventive combined extract prepared in Examples has potent inhibitory effect on inflammatory response.
- LPS lipopolysaccharide
- the level of IL-10, an inflammatory cytokine, in the collected cell supernatant solution was measured.
- test samples were treated in THP-1 cells for four hours and LPS was treated therewith to confirm the anti-inflammatory effect of the test sample.
- IL-10 a pro-inflammatory cytokine
- test sample prepared in Examples sharply reduced the level of IL-10 which had been increased with the dose-dependently increased LPS.
- the inventive combined extract prepared in Examples has potent inhibitory effect on inflammatory response.
- LPS lipopolysaccharide
- LPS lipopolysaccharide
- the expressed level of Beclin 1 and LC3B was quantified by scanning the resulting photo-sensitized film with ChemiDoc TM MPImagingSystem (Biorad).
- Beclin 1 and LC3B WIN1001X ⁇ g/mL LPS 20 ug/mL LPS 100 ug/mL ratio Beclin/ ⁇ -actin ratio LC3/ ⁇ -actin ratio Beclin/ ⁇ -actin ratio LC3/ ⁇ -actin 0 0.67 1.18 0.81 1.35 10 0.76 0.86 0.63 1.59 25 0.37 1.05 0.82 2.49 50 0.41 2.55 1.39 2.28 100 0.25 3.47 1.10 4.57
- Rats (Male Wista rats, Central Lab. Animal, Seoul, Korea, 7 to 8 weeks aged, 200 to 250g) were bred in the well-controlled breeding room in polysulfone cage (2-3 mice per cage) maintaining the temperature of 21 ⁇ 2°C and relative humidity of 50 ⁇ 20% with the light cycle of day/dark (08:00 ⁇ 20:00) at the interval of 12 hours and acclimated to the surround environment
- Test method 3mg/kg of Monosodium iodoacetate (MIA, I2512, Sigma, Poole, UK) was dissolved in the injection saline to re reach to 60 mg/ml concentration on the day of the experiment (day 0). After dividing each groups, the experimental animals were placed in the anesthesia chamber and anesthetized with diethyl ether. 50 ⁇ L of MIA (3 mg/body) was injected into the right knee joint through the infrapatellar ligament using a 1cc syringe (26.5 gauge) in order to inducing osteoarthritis
- MIA Monosodium iodoacetate
- test groups See Table 8.
- test sample treatment groups (G2, G3) were prepared by homogenizing the test samples with vehicle (saline) according to the prescribed dose and orally administrating 1 mL of test sample once a day.
- the negative control group (G1) was treated with only vehicle and orally administrated once a day according to the sample schedule as the test substance administration.
- the positive control group (G4) was homogenized to the vehicle according to the prescribed dose and orally administered once a day.
- the pain threshold (nociceptive latency, threshold) was determined by using dynamic plantar aesthesiometer (Ugo Basile, 37400, Comerio, Italy), a device that gradually increases the force on the feet of Male Wistarats over a certain period of time, according to a von Freestyle evaluation method to measure pain thresholds ((Kwon JY et al., Sci Rep. 2018 Sep 14;8(1):13832)
- the animal Prior to measurement, the animal was placed in an acrylic box with a wire mesh bench and acclimated for five minutes.
- the pain threshold was determined by measuring the weight showing paw withdrawal behavior which animal withdraws its feet by applying a slow force of 0 to 50 g over 10 seconds using metal filaments in the center of each animal's right hind foot.
- the cut-off threshold was set to 50 g, and the measurement time was set to the day before the inducing day of osteoarthritis by the treatment of MIA to calculate base line value in the normal control group (G1), test sample groups (G2, G3) and positive control group (G4) and the same value was obtained in the base line (day3).
- the test result of pain threshold was calculated according to (a) paw withdrawal latencies (sec) and (b) paw withdrawal threshold (g).
- test sample group treated with100 and 150 mg/kg of test samples showed potent inhibitory effect on pain comparing with negative control group treated with vehicle, dose-dependently manner.
- MIA inducement vehicle WIN1001X 100 mg/kg (s) WIN1001X 150 mg/kg (s) Celecoxib 30 mg/kg (s) Before MIA treatment 16.8 ⁇ 0.8 16.2 ⁇ 0.6 16.5 ⁇ 0.6 16.0 ⁇ 1.5 3 days after MIA treatment 9.4 ⁇ 1.2 8.0 ⁇ 1.1 9.7 ⁇ 1.1 9.2 ⁇ 2.1 6 days after MIA treatment 8.6 ⁇ 1.0 11.0 ⁇ 1.3 11.2 ⁇ 1.3 9.9 ⁇ 2.2 11 days after MIA treatment 9.1 ⁇ 1.4 12.4 ⁇ 2.0 11.4 ⁇ 1.8 10.2 ⁇ 0.9 13 days after MIA treatment 9.0 ⁇ 1.6 11.4 ⁇ 1.0 12.1 ⁇ 1.5 10.4 ⁇ 0.9 17 days after MIA treatment 9.5 ⁇ 0.4 11.1 ⁇ 0.4 13.1 ⁇ 1.4 10.9 ⁇ 1.8 21 days after MIA treatment 9.0 ⁇ 1.6 12.8
- MIA inducement vehicle WIN1001X 100 mg/kg (g) WIN1001X 150 mg/kg (g) Celecoxib 30 mg/kg (g) Before MIA treatment 33.6 ⁇ 1.7 32.5 ⁇ 1.2 33.0 ⁇ 1.2 33.8 ⁇ 2.9 3 days after MIA treatment 19.0 ⁇ 2.4 16.2 ⁇ 2.1 19.7 ⁇ 2.2 18.6 ⁇ 4.1 6 days after MIA treatment 17.4 ⁇ 1.8 22.3 ⁇ 2.7 22.6 ⁇ 2.5 19.9 ⁇ 4.3 11 days after MIA treatment 18.5 ⁇ 2.8 25.0 ⁇ 4.0 23.1 ⁇ 3.6 20.7 ⁇ 1.7 13 days after MIA treatment 18.3 ⁇ 3.2 23.0 ⁇ 2.0 24.5 ⁇ 2.9 21.0 ⁇ 1.6 17 days after MIA treatment 19.2 ⁇ 0.8 22.5 ⁇ 0.9 26.3 ⁇ 2.7 22.0 ⁇ 3.4 21 days after MIA treatment 18.3 ⁇ 3.1 2
- test animal shall be correctly located in the holder and fixed so that both feet can be stepped symmetrically, since the load can be varied depending on the position of the animal's foot.
- the particular person performs the fixation to reduce the possible error to the maximum extent.
- the mean value for each test result was digitized into the weight bearing(g) of each foot.
- test sample group potently improved the balance capability of both limbs comparing with the negative control group.
- MIA inducement vehicle WIN1001X 100 mg/kg (%) WIN1001X 150 mg/kg (%) Celecoxib 30 mg/kg (%) Before MIA inducement 50.6 ⁇ 1.2 49.6 ⁇ 0.8 50.3 ⁇ 1.3 49.3 ⁇ 0.9 3 days after MIA inducement 36.9 ⁇ 1.8 37.9 ⁇ 2.7 35.2 ⁇ 2.4 34.7 ⁇ 4.0 6 days after MIA inducement 35.5 ⁇ 1.5 38.3 ⁇ 1.7 38.6 ⁇ 1.3 38.5 ⁇ 1.8 11 days after MIA inducement 33.8 ⁇ 2.9 41.0 ⁇ 2.5 39.1 ⁇ 3.0 40.0 ⁇ 3.2 13 days after MIA inducement 32.3 ⁇ 2.8 43.6 ⁇ 1.4 44.2 ⁇ 1.5 44.5 ⁇ 0.7 17 days after MIA inducement 32.2 ⁇ 1.7 42.9 ⁇ 1.2 42.5 ⁇ 1.3 43.9 ⁇ 1.3 21 days after MIA inducement 31.6 ⁇ 2.0 40.6 ⁇ 1.2 41.0 ⁇ 1.5
- the MIA and test sample were administered to the right knee joint of sacrificed rats (male Wistar rats) and fixed with formalin.
- the degree of bone injury around femur area of the knee joint in each group consisting of 3 rats was determined using X-ray source (70kV, 142uA, AI 0.5mm filter, rotation step 0.6 °) and animal scanner (SKYSCAN1272 ex-vivo micro-CT, Bruker micro CT, Belgium) to perform micro-CT photography at 15 ⁇ m of Pixel resolution including section formation (NRecon), section rotation (Data Viewer), data Analysis (CTAN), Volume rendering generation (CTVox), and surface rendering (CTAN+CTVol).
- test sample group treated with100 and 150 mg/kg of test samples showed potent inhibitory effect on bone injury comparing with negative control group (vehicle group) through the test result of Micro-CT photography.
- the MIA and test sample were administered to the right knee joint of sacrificed rats (male Wistar rats) and the right knee joint of the rat fixed with formalin was sliced to stain with Safranin O.
- the histopathological analysis was performed by photographing the femur area of knee joint (x200 fold). The test result was calculated and evaluated according to the known methods including Total Mankin score (Bulstra SK et al., 1989, Clin Orthop Relat Res: 294-302.) and OARSI score (Pritzker K.P.H. et al., 2006, Osteoarthritis Cartilage 14:13-29).
- test sample group treated with100 and 150 mg/kg of test samples showed potent inhibitory effect on bone injury comparing with negative control group (vehicle group) through the test result of histopathological analysis
- Injection preparation was prepared by dissolving active component, controlling pH to about 7.5 and then filling all the components in 2ml ample and sterilizing by conventional injection preparation method.
- Powder preparation was prepared by mixing above components and filling sealed package.
- Tablet preparation was prepared by mixing above components and entabletting.
- Lactose 50mg
- Tablet preparation was prepared by mixing above components and filling gelatin capsule by conventional gelatin preparation method.
- Liquid preparation was prepared by dissolving active component, and then filling all the components in 1000ml ample and sterilizing by conventional liquid preparation method.
- Vitamin mixture optimum amount
- Vitamin E 1.0mg
- Vitamin B 10 13mg
- Vitamin B 2 0.15mg
- Vitamin B6 0.5mg
- Vitamin B1 20.2g
- Vitamin C 10mg
- Zinc oxide 0.82mg
- Citric acid 1000mg
- Health beverage preparation was prepared by dissolving active component, mixing, stirred at 85°C for 1 hour, filtered and then filling all the components in 1000ml ample and sterilizing by conventional health beverage preparation method.
- the present invention provides an oral pharmaceutical composition
- the present inventors demonstrated that the anti-inflammatory/anti-arthritic effects of inventive combined composition is potent by accomplishing in vitro experiments such as the inhibitory test on the expression of cytokines involved in inflammation (RPLPO, TSLP, GM-CSF and IL-1beta) (Experimental Example 1); Cell viability test on HT-29 and THP-1 cell ( in vitro Experimental Example 2); Anti-inflammatory activity in THP-1 cell ( in vitro, Experimental Example 3); inhibitory effect on autophagy activity ( in vitro, Experimental Example 4.) as well as in vivo experiments such as inhibitory effect on arthritis using by arthritis-induced rat animal model ( in vivo, Experimental Example 5), therefore, it is confirmed that inventive combined extract is very useful in the alleviation or treatment of inflammatory disease and arthritis disease as a form of oral pharmaceutical composition.
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Botany (AREA)
- Epidemiology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Medical Informatics (AREA)
- Alternative & Traditional Medicine (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Nutrition Science (AREA)
- Immunology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Rheumatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Physical Education & Sports Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention relates to an oral pharmaceutical composition comprising a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix, the present inventors demonstrated that the anti-arthritic effect of inventive combined composition is potent by accomplishing in vitro experiments such as the inhibitory test on the expression of cytokines involved in inflammation (RPLPO, TSLP, GM-CSF and IL-1beta) (Experimental Example 1); Cell viability test on HT-29 and THP-1 cell (in vitro Experimental Example 2); Anti-inflammatory activity in THP-1 cell (in vitro, Experimental Example 3); inhibitory effect on autophagy activity (in vitro, Experimental Example 4.) as well as in vivo experiments such as inhibitory effect on arthritis using by arthritis-induced rat animal model (in vivo, Experimental Example 5), therefore, it is confirmed that inventive combined extract is very useful in the alleviation or treatment of inflammatory disease and arthritis disease as a form of oral pharmaceutical composition.
Description
The present invention is related to an oral pharmaceutical composition comprising an extract of combined herbs comprising Longanae arillus for the treatment or prevention of arthritis diseases and the use thereof.
Generally, an inflammatory response is a normal response of human body associated with an edema, a pain etc in case that a tissue or a cell received any invasion causing some organic change in the tissue or cell. Recently, various kinds of cytokines have been found to be involved in the inflammatory disease.
Therefore, many studies have been performed to develop effective drugs to inhibit the production of various cytokines such as IL-4, IL-5, and IL-13, and immunoglobulin E etc, which are involved in the activation of inflammatory cells resulting in leucotriene biosynthesis, a system secreted by inflammatory cells such as neutrophil etc and caused by the occurrence of inflammation, allergic response or asthma etc.
While the inflammatory phase continues to develop with pro-inflammatory cytokines such as TNF-α, IL-1β, IL-6 etc and MMPs (matrix metalloproteinases, such as PDGF, VEGF, and IGF, it is known that the expression of growth factor reduces (Trengove NJ, Bielefeldt-Ohmann H, Stacey MC (2001) Mitogenic activity and cytokine levels in non-healing and healing chronic leg ulcers. Wound Repair and Regeneration. 8: 13-25.; Armstrong DG, Jude EB (2002) The Role of Matrix Metalloproteinases in Wound Healing. Journal of the American Podiatric Medical Association. 92: 12-18.).
MMPs (matrix metalloproteinases) are controlled by TIMPs (tissue inhibitors of metalloproteinases) in wound area, which decompose extracellular substrates, enabling re-epithelialization (Martins VL, Caley M, O’ Toole EA (2013) Matrix metalloproteinases and epidermal wound repair. Cell and Tissue Research. 351: 255-268).
In particular, there have been focused in the research on MMP-9 which has been known to have the most harmful effects on chronic wounds among MMPs (Jones JI, Nguyen TT, Peng Z, Chang M (2019) Targeting MMP-9 in Diabetic Foot Ulcers. Pharmaceuticals. 12: 79.; Reiss MJ, Han YP, Garcia E, Goldberg M, Yu H, Garner WL (2010) Matrix metalloproteinase-9 delays wound healing in a murine wound model. Surgery. 147: 295-302.).
Accordingly, there has been still needed to develop more effective drug and cosmetics in treating and alleviating arthritis diseases from natural source with low side effects than conventionally used drugs till now.
Longanae Arillus, a seed coat of Dimocarpus longan, Euphoria longan or the same species belonged to Sapindaceae has been reported to contain a glucose, fructose, protein etc and to show cardio-protective effect, appetite stimulanting effect etc (Chung B. S et al, Dohaehyangyakdaesajeon, youngrimsa, 2nd Ed. p197-198, 1998).
Ligustici Tenuissimi Rhizoma, a rhizoma or root of Ligusticum tenuissimum Kitagawa, Ligusticum sinense Oliv, Ligusticum jeholense Nakai et Kitagawa or the same species belonged to Umbelliferae has been reported to contain a cnidilide, 3-butyl phthalide etc and to show anti-bacterial effect etc (Chung B. S et al, Dohaehyangyakdaesajeon, youngrimsa, 2nd Ed. P428-429, 1998).
Polygalae radix, a root of Polygala tenuifolia Willd., or the same species belonged to Polygalaceae has been reported to contain various sanponis and to show expectorant activity, anti-bacterial effect etc (Chung B. S et al, Dohaehyangyakdaesajeon, youngrimsa, 2nd Ed. P798-799, 1998).
However, there has been not reported or disclosed on the preventing or alleviating activity of an orally applied extract of combined herbs of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix showing potent treating effect on arthritis diseases in any of above cited literatures, and the disclosures of which are incorporated herein by reference.
To investigate the anti-rheumatic effect of a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix, the inventors of present invention have intensively carried out various experiments including in vitro experiments such as the inhibitory test on the expression of cytokines involved in inflammation (RPLPO, TSLP, GM-CSF and IL-1beta) (Experimental Example 1); Cell viability test on HT-29 and THP-1 cell (in vitro Experimental Example 2); Anti-inflammatory activity in THP-1 cell (in vitro, Experimental Example 3); inhibitory effect on autophagy activity (in vitro, Experimental Example 4.) as well as in vivo experiments such as inhibitory effect on arthritis using by arthritis-induced rat animal model (in vivo, Experimental Example 5). As a result of these investigations, the inventors finally completed the present invention by confirming that inventive combined herb extract strongly inhibited and alleviated arthritis diseases.
The technical solution to solve the problem of the background art is for the development of novel herb formulation for treating and preventing inflammation disease or arthritis diseases.
Accordingly, it is an object of the present invention to provide an oral pharmaceutical composition comprising a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix as an active ingredient to treat and alleviate arthritis diseases.
The term “combined herb extract” defined herein comprises the combined herb extract, i.e., (a) combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix with the mixed ratio based on the dried weight of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix (w/w) ranging from 0.01 -100: 0.01 -100: 0.01 -100 weight part (w/w), preferably, 0.1-50: 0.1-50: 0.1-50 weight part (w/w), more preferably, 0.1-10: 0.1-10: 0.1-10 weight part (w/w), more and more preferably, 1-5: 1-5: 1-5 weight part (w/w), most preferably, 1-3: 1-3 : 1-3 weight part (w/w); or (b) the combination of each extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix with the mixed ratio based on the dried weight of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix (w/w) ranging from 0.01 -100: 0.01 -100: 0.01 -100 weight part (w/w), preferably, 0.1-50: 0.1-50: 0.1-50 weight part (w/w), more preferably, 0.1-10: 0.1-10: 0.1-10 weight part (w/w), more and more preferably, 1-5: 1-5: 1-5 weight part (w/w), most preferably, 1-3: 1-3 : 1-3 weight part (w/w) in the present invention.
The term “extract” disclosed herein comprises the extract which can be extracted with at least one solvent selected from water, C1-C4 lower alkyl alcohol such as methanol, ethanol, propanol, butanol, etc, acetone, ethyl acetate, chloroform, hexane, butyleneglycol, propyleneglycol or glycerin, preferably, water, methanol, ethanol, more preferably, water or 10-90 %(v/v) ethanol in water, most preferably, water or 20-80 %(v/v) ethanol in water.
The term “arthritic diseases” disclosed herein comprises the disease selected from rigid spondylitis, degenerative arthritis, osteoarthritis, rheumatoid arthritis, gout or frozen shoulder etc.
The term, “anti-Inflammation” disclosed herein, not limited thereto, means all the mechanism to inhibit various inflammation.
The term, “anti-arthritic” disclosed herein, not limited thereto, means all the mechanism to inhibit various rheumatism.
Inflammation is part of the complex biological response of body tissues to harmful stimuli, such as pathogens, damaged cells, or irritants, and the nonspecific immune response such as heat, pain, redness, swelling, etc is called as “inflammatory response”.
Inflammation can be classified as (a) acute inflammation, the initial response of the body to harmful stimuli, is achieved by the increased movement of plasma and leukocytes (especially granulocytes) from the blood into the injured tissues and then a series of biochemical events propagates and matures the inflammatory response, involving the local vascular system, the immune system, and various cells within the injured tissue and (b)Prolonged inflammation, known as chronic inflammation, leading to a progressive shift in the type of cells present at the site of inflammation, such as mononuclear cells, and being characterized by simultaneous destruction and healing of the tissue from the inflammatory process.
Generally, the macrophage in damaged cell excrete various cytokines, which activates T lymphocyte and mast cell, a lymphocyte, releases various histamines, which initiate internal barrier response, resulting in inducing inflammation of the inflected cells. Accordingly, the expressed level of cell cytokines may be used as an indicator of the activation of inflammatory response (the other aspects, anti-inflammatory activity). The “anti-inflammatory activity” disclosed herein denotes the inhibitory activity against various skin inflammation.
Cytokines means all the immunological substances including chemokines, interferons, interleukins, lymphokines, and tumour necrosis factors produced by a broad range of cells, including immune cells like macrophages, B lymphocytes, T lymphocytes and mast cells, as well as endothelial cells, fibroblasts, and various stromal cells, which are released through immunological progress caused by the infiltration of various pathogen such as virus etc.
Generally, cytokines are released at initial stage of infection, however, released constantly where the immune system becomes extraordinarily activated. When the high-level of cytokines are released for a long time such as more than week, we called as “a cytokine storm”, which is a physiological reaction in which the innate immune system causes an uncontrolled and excessive release of pro-inflammatory signaling molecules called cytokines and it exacerbates the inflammation resulting from the extremely abundant homing of immune cells to the inflected area, causes to blood extravasation through the loosening of blood vessel and severely to death. The term “the inhibitory activity of cytokine expression” disclosed herein can be interpreted as a prevention, treatment or improvement of cytokine storm.
The term “cytokine” disclosed herein, not intended to limit thereto, comprises various cytokine involved in dermatitis, such as atopic dermatitis, specifically, the cytokine selected from group of TLSP (thymic stromal lymphopoietin), colony stimulating factor (CSF) such as GM-CSF (granulocyte-macrophage colony stimulating factor), M-CSF (macrophage colony stimulating factor), G-CSF (granulocyte colony stimulating factor) and the like, interleukins such as interleukin-1 (IL-1), IL-4, IL-10, IL-12, IL-13, IL-31, IL-33 and the like, tumor necrosis factor alpha (TNF-α), interferon gamma (IFNγ) etc,
An inventive extract may be prepared in accordance with the following preferred embodiment.
For the present invention, above described extract can be prepared by follows;
The term “combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix” defined herein can be prepared by the procedure comprising the steps; of slicing and washing Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix” to use as a basic extraction material at 1st step; mixing together thoroughly with the mixed ratio based on the dried weight of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix (w/w) ranging from 0.01 -100: 0.01 -100: 0.01 -100 weight part (w/w), preferably, 0.1-50: 0.1-50: 0.1-50 weight part (w/w), more preferably, 0.1-10: 0.1-10: 0.1-10 weight part (w/w), more and more preferably, 1-5: 1-5: 1-5 weight part (w/w), most preferably, 1-3: 1-3 : 1-3 weight part (w/w) to afford the mixed material at 2nd step; adding 1 - 20 fold volume(v/w), preferably, 4 -8 fold volume(v/w) of extracting solvent selected from the group consisting of water, C1-C4 lower alkyl alcohol such as methanol, ethanol, propanol, butanol, etc, acetone, ethyl acetate, chloroform, hexane, butyleneglycol, propyleneglycol or glycerin, preferably, water, methanol, ethanol, more preferably, water or 10-90 %(v/v) ethanol in water, most preferably, water or 20-80 %(v/v) ethanol in water to the mixed material at 3rd step; extracting each solution with the extraction method by the extraction with hot water extraction, cold water extraction, reflux extraction or ultra-sonication extraction, preferably, hot water extraction at the temperature ranging from 50℃ to 120℃, preferably, about 80℃ to 100℃, for the period ranging from 1 to 24 hours, preferably, 2 to 12 hours at 4th step; repeating the above-described extraction process to collect each filtrate with filtration, drying through freeze drying, natural air drying or hot air drying process, preferably freeze drying process to obtain the combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix of the present invention.
It is another object of the present invention to provide a process for preparing the combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix of the present invention.as described above.
It is another object of the present invention to provide an oral pharmaceutical composition comprising a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix prepared by the above-described process, as an active ingredient to treat and alleviate arthritis diseases.
In accordance with another aspect of the present invention, there is also provided a method of treating or alleviating arthritis diseases in a mammal comprising orally administering to said mammal an effective amount of the combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix and pharmaceutically acceptable carrier thereof.
In accordance with the other aspect of the present invention, there is also provided a use of the combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix for manufacture of oral preparation employed for treating or alleviating arthritis diseases in mammals including human as an active ingredient.
It is still another object of the present invention to provide a pharmaceutical composition or health functional food comprising the herb extract of the above-mentioned herb obtained by the above described process as an active ingredient for preventing and treating arthritis diseases.
The present inventors demonstrated that the anti-inflammatory/anti-arthritic effect of inventive composition is potent by accomplishing in vitro experiments such as the inhibitory test on the expression of cytokines involved in inflammation (RPLPO, TSLP, GM-CSF and IL-1beta) (Experimental Example 1); Cell viability test on HT-29 and THP-1 cell (in vitro Experimental Example 2); Anti-inflammatory activity in THP-1 cell (in vitro, Experimental Example 3); inhibitory effect on autophagy activity (in vitro, Experimental Example 4.) as well as in vivo experiments such as inhibitory effect on arthritis using by arthritis-induced rat animal model (in vivo, Experimental Example 5), therefore, it is confirmed that inventive combined extract is very useful in the alleviation or treatment of inflammatory disease and arthritis disease as a form of oral pharmaceutical composition.
The pharmaceutical composition for treating purposed diseases could contain about 0.01 to 99 w/w% the above herb extract of the present invention based on the total weight of the composition.
However, the amount and each component of the above-mentioned composition can be varied with the patient’s condition, development of patient’s disease, the sort of disease etc.
The inventive composition may additionally comprise conventional carrier, adjuvants or diluents in accordance with a using method.
The herb composition according to the present invention can be formulated in oral dosage form such as powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol and the like; topical preparation; or injection solution. The herb composition according to the present invention can be provided as a pharmaceutical composition containing pharmaceutically acceptable carriers, adjuvants or diluents, e.g., lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starches, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, magnesium stearate and mineral oil. The formulations may additionally include excipients such as fillers, bulking agents, binders, wetting agents, disintegrating agents, surfactants, diluents and the like. The solid oral dosage form comprises tablet, pill, powder, granule, capsule and the like and the solid oral dosage form is prepared by adding at least one excipient such as starch, calcium carbonate, sucrose, lactose or gelatin and the like to the herb extract. Lubricant such as magnesium stearate or talc may be used. The aqueous oral dosage form comprises suspension, oral solution, emulsion, syrup and the aqueous oral dosage form may comprise several excipients such as wetting agents, sweetener flavoring agents, preservatives, as well as water, liquid paraffin. The parenteral dosage form comprises sterilized aqueous solution, non-aqueous solvent, suspension, emulsion, lyophilized preparation, suppository, and the like. Suitable examples of the carriers include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable ester such as ethyl oleate. Base for suppository may include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatine etc., but are not limited to them.
The desirable dose of the inventive composition varies depending on the condition and the weight of the subject, severity, drug form, route and period of administration, and may be chosen by those skilled in the art. However, in order to obtain desirable effects, it is generally recommended to administer at the amount ranging 0.01mg/kg to10g/kg, preferably, 1mg/kg to 1g/kg by weight/day of the inventive composition of the present invention. The dose may be administered in a single or multiple doses per day.
The pharmaceutical composition of present invention can be administered to a subject animal such as mammals (rat, mouse, domestic animals or human) via various routes. All modes of administration are contemplated, for example, administration can be made orally, rectally or by intravenous injection.
In accordance with one aspect of the present invention, there provided a health functional food comprising a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix, as an active ingredient to prevent or improve arthritis diseases.
The term "a health functional food" defined herein comprises the functional food having enhanced functionality such as physical functionality or physiological functionality by adding the extract of the present invention to conventional food to prevent or improve the purposed diseases in human or mammal and stipulated by the Law for Health Functional Foods 6727 in Republic of Korea.
The health functional food composition for preventing and improving purposed diseases could contain about 0.01 to 95 w/w%, preferably 1 to 80 w/w% of the above herb composition of present invention based on the total weight of the composition.
Moreover, the inventive extract of the present invention also can be used as a main component or additive and aiding agent in the preparation of various functional health food and health supplement food for the prevention or improvement of arthritis diseases.
The inventive health functional food may be prepared and processed by the form of pharmaceutically acceptable dosage form such as powder, granule, tablet, capsule, pills, suspension, emulsion, syrup and the like; or the functional health food form such as tea bag, leached tea, health beverage type and the like.
It is the other object of the present invention to provide a health supplement food comprising a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix, as an active ingredient or as a main component to prevent or improve arthritis diseases.
The above-mentioned term “as a main component” means that the above health supplement food comprises about 30 to 99 (w/w%), preferably 50 to 99 (w/w%), more preferably 70 to 99 (w/w%) of the inventive extract of present invention based on the total weight of the composition.
When the combined herb extract of the present invention is used as a component in the health functional beverage composition, the health functional beverage composition can comprise other component such as flavoring agent or natural carbohydrate without limits like that typical beverage composition. Examples of the natural carbohydrate comprise monosaccharide such as glucose, fructose etc; disaccharide such as maltose, sucrose etc; and polysaccharide, for example, sugar such as dextrin, cyclodextrin, and sugar alcohol such as xylitol, sorbitol, erythritol. Natural flavoring agent (thaumatin, stevia extract (rebaudioside A, glycyrrhizin, etc)) and synthetic flavoring agent (saccharin, aspartame, etc) may be added in the health functional beverage composition. The amount of natural carbohydrate generally ranges from about 1 to 20g, preferably about 5 to 12g per 100ml of the present composition.
When the combined herb extract of the present invention is used as a food additive of the health food, the combined herb extract may be added intact or used with other food ingredient according to general process. Examples of the food comprises meat products, sausage, bread, chocolate, candy, snack, cracker, biscuit, pizza, ramen, noodle products, chewing gum, dairy products such as ice cream, soup, beverage, tea, drinks, alcohol drink, vitamin complex etc, but not intended herein to limit thereto, for preventing or improving of purposed disease.
The other components than aforementioned composition are various nutrients, a vitamin, a mineral or an electrolyte, synthetic flavoring agent, a coloring agent and improving agent in case of cheese, chocolate et al., pectic acid and the salt thereof, alginic acid and the salt thereof, organic acid, protective colloidal adhesive, pH controlling agent, stabilizer, a preservative, glycerin, alcohol, carbonizing agent used in carbonate beverage et al. The other component than aforementioned ones may be fruit juice for preparing natural fruit juice, fruit juice beverage and vegetable beverage, wherein the component can be used independently or in combination. The ratio of the components is not so important but is generally range from about 0 to 20 w/w % per 100 w/w % present composition.
Also, above described extract can be added to food or beverage for prevention and improvement of purposed disorder. The amount of above described extract in food or beverage as a functional health food or health supplement food may generally range from about 0.01 to 15 w/w % of total weight of food for functional health food composition. And the extract of the present invention may be added 0.02 to 5g, preferably 0.3 to 1g per 100ml in health beverage composition.
As described in the present invention, the present inventors demonstrated that anti-inflammatory/anti-arthritic effect of inventive combined composition is potent by accomplishing in vitro experiments such as the inhibitory test on the expression of cytokines involved in skin inflammation (RPLPO, TSLP, GM-CSF and IL-1beta) (Experimental Example 1); Cell viability test on HT-29 and THP-1 cell (in vitro Experimental Example 2); Anti-inflammatory activity in THP-1 cell (in vitro, Experimental Example 3); inhibitory effect on autophagy activity (in vitro, Experimental Example 4.) as well as in vivo experiments such as inhibitory effect on arthritis using by arthritis-induced rat animal model (in vivo, Experimental Example 5), therefore, it is confirmed that inventive combined extract is very useful in the alleviation or treatment of inflammatory disease and arthritis disease as a form of oral pharmaceutical composition.
It will be apparent to those skilled in the art that various modifications and variations can be made in the compositions, use and preparations of the present invention without departing from the spirit or scope of the invention.
The present invention is more specifically explained by the following examples. However, it should be understood that the present invention is not limited to these examples in any manner.
EXAMPLES
The following Examples and Experimental Examples are intended to further illustrate the present invention without limiting its scope.
Example 1. The preparation of inventive combined extract (1)
20g of dried Longanae Arillus (Buyoung Yakup Co. Ltd.), 20g of dried Ligustici Tenuissimi Rhizoma (Buyoung Yakup Co. Ltd.) and 20g of dried Polygalae radix(Buyoung Yakup Co. Ltd.) were cut into small pieces, mixed with 6 fold volume (v/w) of 20% ethanol in water and the mixture was subjected to reflux extraction at 90 ± 5℃ for 3 days. After filtration of the extract through filter paper (pore size, less than 10㎛) to remove the debris, the remaining debris was further extracted two times with 4 fold volume (v/w) of 20% ethanol in water and the extract was filtered with filter paper (pore size, less than 10㎛).
The collected extract was mixed with together and concentrated under vaccuo (16-21 brix) to afford concentrated extract. The concentrated extract was dried with freeze drying process and pulverized (less than 50 mesh) to obtain 20.5 g (powder as dried basis, yield 33.4%) of inventive combined extract (1) (designated as “WIN-1001X” hereinafter)
Example 2-6. The preparation of inventive combined extract (2)-(6)
Excepting adopting different combined ratio as well as different solvents disclosed in Example 1, all the procedure was identical with those in Example 1 to obtain various inventive combined extract of Longanae Arillus (LA) , Ligustici Tenuissimi Rhizoma (LT) and Polygalae radix (PR) i.e., inventive combined extract (2) to inventive combined extract (6) of the present invention, which are used as a test samples in following experiment.
Sample weight (g) | |||||||
Example | LA* | PR* | LT* | solvent* | name | Extract weight | Final yield |
Example 2 | 10 | 5 | 50 | 10% EtOH | WIN-1002X | 16.6g | 25.6% |
Example 3 | 20 | 50 | 5 | Water | WIN-1003X | 24.7g | 32.9% |
Example 4 | 10 | 80 | 20 | 70% BuOH | WIN-1004X | 32.3g | 29.4% |
Example 5 | 5 | 50 | 20 | 50% EtOH | WIN-1005X | 21.5g | 28.7% |
Example 6 | 30 | 10 | 2 | hexane | WIN-1006X | 12.6g | 30.1% |
*: Longanae Arillus (LA) , Ligustici Tenuissimi Rhizoma (LT), Polygalae radix (PR) |
Experimental Example 1. inhibitory effect on cytokine expression (
in vitro
).
In order to determine the anti-inflammatory activity of inventive extract, following inhibition test of cytokine expression using HaCaT cell was performed according to the procedure disclosed in the literature (Jeong et al., 2019, J. Invest. Dermatol., May; 139 (5): pp1098-1109).
HaCaT cell (human epithelial keratinocyte cell, 300493, CLS) was inoculated into DMEM medium containing 10% Fetal bovine serum, 100units/ml of penicillin, 100μg/ml of streptomycin (D6429, Sigma-Aldrich Co. Ltd) and was incubated in the incubator (HERA cell 150i, Thermo Fisher Scientific Co. Ltd.) maintaining optimum humidity (85-95%) and 5% CO2 atmosphere.
For performing gene expression test, the incubated cells were transferred to 12 wells and 50ng/ml of TNF alpha (RC214-12, Biobasic Co. Ltd) was treated therewith for 1 hour to induce inflammatory response. Dexamethasone (200nM, positive control, “DEX”, D4902, Sigma-Aldrich Co. Ltd.) and distilled water (negative control, “DIW”) were used as comparative controls.
1 hour after inducing the inflammation, 1μg/ml of inventive extract prepared in Examples was treated with identical medium and subjected to incubation for 1 hour. After the incubation, RNA (FATRR-001, Favorgen) was extracted from the cell and cDNA was synthesized from the RNA using by cDNA synthesis kit (RRO36A, TAKARA). The polymerization reaction was performed using by the synthesized cDNA and Sybrgreen kit (RT500M, Enzynomics) and then Real-time-PCR was performed using by primers for various cytokines involved in skin inflammation (RPLPO, TSLP, GM-CSF and IL-1beta) as disclosed in Table 2.
human* | direction | sequence | Sequence I. D |
RPLP0 | forward | 5'- AGC CCA GAA CAC TGG TCT C-3' | 1 |
reverse | 5’- ACT CAG GAT TTC AAT GGT GCC-3’ | 2 | |
TSLP | forward | 5'-TAT GAG TGG GAC CAA AAG TAC CG-3' | 3 |
reverse | 5'-GGG ATT GAA GGT TAG GCT CTG G-3' | 4 | |
GM-CSF | forward | 5'-TCC TGA ACC TGA GTA GAG ACA C-3' | 5 |
reverse | 5'-TGC TGC TTG TAG TGG CTG G-3' | 6 | |
IL-1β | forward | 5'-CTC CAG GGA CAG GAT ATG GA-3' | 7 |
reverse | 5'-TCT TTC AAC ACG CAG GAC AG-3' | 8 | |
* : abbreviation- RPLP0 (Ribosomal Protein Lateral Stalk Subunit P0); TSLP (thymic stromal lymphopoietin); GM(Granulocyte-macrophage)-CSF (colony stimulating factor); IL (interleukin) |
As can be seen in Table 3 showing quantitative result of the RT-PCR, the test sample group treated with the inventive extract, sharply inhibited the expressed level of various cytokine involved in inflammation comparing with negative control group treated with distilled water (DIW) and it has been confirmed that the inhibitory activity of the test sample on the expression of various cytokine involved in inflammation is equivalent to that of positive control group treated with dexamethasone (DEX).
Accordingly, it has been confirmed that the various kind of inventive combined extract prepared in Examples 1-6 have potent inhibitory effect on inflammation.
TSLP | ||||||
- - |
TNFα DIW |
TNFα WIN-1001X |
TNFα WIN-1002X |
TNFα WIN-1003X |
TNFα WIN-1005X |
TNFα Dex |
1 | 132.4692 | 47.43735 | 60.85783 | 48.9323 | 55.34286 | 52.49334 |
0.462769 | 26.91228 | 9.645089 | 24.95619 | 19.85678 | 26.59252 | 11.2336 |
GM-CSF | ||||||
- - |
TNFα DIW |
TNFα WIN-1001X |
TNFα WIN-1002X |
TNFα WIN-1003X |
TNFα WIN-1005X |
TNFα Dex |
1 | 4.473627 | 1.982161 | 2.069408 | 2.384771 | 1.917569 | 1.935997 |
0.111 | 0.817826 | 0.889233 | 0.326074 | 0.871501 | 0.599711 | 0.581338 |
IL-1β | ||||||
- - |
TNFα DIW |
TNFα WIN-1001X |
TNFα WIN-1002X |
TNFα WIN-1003X |
TNFα WIN-1005X |
TNFα Dex |
1 | 4.152715 | 1.407169 | 1.437399 | 2.064964 | 1.662578 | 1.080503 |
0.483565 | 1.087056 | 0.394622 | 0.1926 | 0.620225 | 0.193175 | 0.413136 |
Experimental Example 2. Cell viability test on HT-29 and THP-1 cell (
in vitro
).
In order to confirm the cytotoxicity of inventive extract on HT-29 cell and THP-1 cell, following cell viability using HT-29 cell and THP-1 cell was according to the previous known procedure in the art.
2-1. Procedure
HT-29 cell (human colon epithelial cell, Korean Cell Line Bank, Korean Cell Line Research Foundation, 101, Daehak-ro, Jongno-gu, Seoul, 03080, Korea) was inoculated into DMEM medium containing 10% Fetal bovine serum and 1% penicillin-streptomycin solution and THP-1 cell (human monocyte cell, Korean Cell Line Bank, Korean Cell Line Research Foundation, 101, Daehak-ro, Jongno-gu, Seoul, 03080, Korea) was inoculated into RPMI medium containing 10% Fetal bovine serum and 1% penicillin-streptomycin solution to incubate.
25, 50, 100, 250, 500 and 1000μg/mL of test samples were added to HT-29 cell and THP-1 cell. 24 hours after the incubation, CCK-8 (cell counting kit-8, Dojindo Molecular Technologies, Inc.) was added thereto and the absorbance (optical density) was determined at 450nm to determine cell viability.
2-2. test result
As can be seen in Table 4, it has been confirmed that the cell viability of test sample group treated with the inventive extract (25-1000 μg/mL) in HT-29 cell showed similar to that of negative control group treated with only medium and that in THP-1 cell showed 90.5% and 26.5% at the concentration of 500 and 1000 μg/mL, respectively, which showed some different with that of negative control group and less than 500 μg/mL of test sample was applied to following test.
WIN1001X ㎍/mL |
HT-29 cell | THP-1 cell |
0 | 100 % | 100 % |
25 | 100.5 ± 0.6 % | 102.2 ± 1.1 % |
50 | 103.3 ± 3.2 % | 103.7 ± 1.5 % |
100 | 105.0 ± 1.4 % | 104.4 ± 1.6 % |
250 | 109.2 ± 3.1 % | 101.1 ± 0.6 % |
500 | 105.9 ± 3.3 % | 90.5 ± 0.8 % |
1000 | 90.3 ± 0.1 % | 26.5 ± 5.8 % |
Experimental Example 3. Anti-inflammatory activity in THP-1 cell (
in vitro
).
In order to determine the anti-inflammatory activity of inventive extract, following inhibition test of the level of pro-inflammatory cytokines using THP-1 cell was performed according to the previous known procedure in the art.
3-1. determination on the level of IL-1beta
3-1-1. test procedure
10, 20, 50, 100, and 500 μg/mL lipopolysaccharide (LPS) was added to human monocyte cell lines (THP-1 cell Korean Cell Line Bank, Korean Cell Line Research Foundation, 101, Daehak-ro, Jongno-gu, Seoul, 03080, Korea) to prepare an inflammatory model.
24 hours after LPS treatment, the level of IL-1beta, an inflammatory cytokine, in the collected cell supernatant solution was measured.
In addition, 10, 25, 50 and 100 μg/mL of test samples were treated in THP-1 cells for four hours and LPS was treated therewith to confirm the anti-inflammatory effect of the test sample.
24 hours after LPS treatment, the level of IL-1beta, a pro-inflammatory cytokine, was measured using ELISA reader (IL-1beta/IL-1F2 Duo set ELISA, R&D Systems).
3-1-2. test result
As can be seen in Table 5, it has been confirmed that the test sample prepared in Examples sharply reduced the level of IL-1beta which had been increased with the dose-dependently increased LPS.
Accordingly, it has been confirmed that the inventive combined extract prepared in Examples has potent inhibitory effect on inflammatory response.
WIN1001X ㎍/mL |
IL-1β level (pg/ mL) | |||
LPS 20 ㎍/mL | LPS 50 ㎍/mL | LPS 100 ㎍/mL | LPS 500 ㎍/mL | |
0 | 117.7 ± 14.0 | 262.8 ± 9.0 | 284.2 ± 9.0 | 319.5 ± 10.4 |
10 | 71.1 ± 6.5 | 214.5 ± 5.5 | 242.1 ± 4.0 | 312.2 ± 9.3 |
25 | 84.9 ± 7.6 | 186.5 ± 6.9 | 213.2 ± 6.2 | 285.1 ± 9.3 |
50 | 68.0 ± 12.4 | 158.4 ± 5.3 | 186.4 ± 3.2 | 245.2 ± 11.0 |
100 | 133.4 ± 8.6 | 198.9 ± 10.3 | 233.9 ± 6.0 | 311.1 ± 8.4 |
3-2. determination on the level of IL-10
3-2-1. test procedure
10, 20, 50, 100, and 500 μg/mL of lipopolysaccharide (LPS) was added to human monocyte cell lines (THP-1 cell Korean Cell Line Bank, Korean Cell Line Research Foundation, 101, Daehak-ro, Jongno-gu, Seoul, 03080, Korea) to prepare an inflammatory model.
24 hours after LPS treatment, the level of IL-10, an inflammatory cytokine, in the collected cell supernatant solution was measured.
In addition, 10, 25, 50 and 100 μg/mL of test samples were treated in THP-1 cells for four hours and LPS was treated therewith to confirm the anti-inflammatory effect of the test sample.
24 hours after LPS treatment, the level of IL-10, a pro-inflammatory cytokine, was measured using ELISA reader (IL-10, Duo set ELISA, R&D Systems).
3-2-2. test result
As can be seen in Table 6, it has been confirmed that the test sample prepared in Examples sharply reduced the level of IL-10 which had been increased with the dose-dependently increased LPS.
Accordingly, it has been confirmed that the inventive combined extract prepared in Examples has potent inhibitory effect on inflammatory response.
WIN1001X ㎍/mL | IL-10 level (pg/ mL) | ||
LPS 50 ㎍/mL | LPS 100 ㎍/mL | LPS 500 ㎍/mL | |
0 | 5.3 ± 1.0 | 12.0 ± 1.8 | 14.2 ± 1.1 |
10 | 6.2 ± 1.6 | 7.3 ± 1.7 | 15.6 ± 1.7 |
25 | 10.6 ± 2.1 | 6.7 ± 1.8 | 19.4 ± 1.8 |
50 | 5.2 ± 1.6 | 14.0 ± 1.6 | 24.5 ± 1.0 |
100 | 45.5 ± 2.5 | 67.3 ± 4.3 | 87.0 ± 4.8 |
Experimental Example 4. inhibitory effect on autophagy activity (
in vitro
).
In order to determine the effect of inventive extract on the expression of Inflammatory factor in immune cell, following test using THP-1 cell was performed according to the previous known procedure in the art.
4-1. test procedure
In order to confirm the correlation between ant-inflammatory activity and autophagy pathway, 10, 20, 50, 100, and 500 μg/mL lipopolysaccharide (LPS) was added to human monocyte cell lines (THP-1 cell Korean Cell Line Bank, Korean Cell Line Research Foundation, 101, Daehak-ro, Jongno-gu, Seoul, 03080, Korea) to prepare an inflammatory model.
After the treatment of 10, 25, 50 and 100 μg/mL of test samples, 20 and 100 μg/mL of lipopolysaccharide (LPS) was added thereto to confirm the expression of Beclin 1 and LC3B (autophagy marker) through immuno-blotting test using anti-Beclin 1 antibody (Abcam) and LC3B-antibody (Cell Signaling), respectively.
The expressed level of Beclin 1 and LC3B was quantified by scanning the resulting photo-sensitized film with ChemiDocTMMPImagingSystem (Biorad).
4-2. test result
As can be seen in Table 7, it has been confirmed that the test group treated with test samples and 100 μg/mL of lipopolysaccharide (LPS), showed the dose-dependently increasing effect on the expression of LC3B-1 and Beclin1.
WIN1001X ㎍/mL | LPS 20 ug/mL | LPS 100 ug/mL | ||
ratio Beclin/β-actin |
ratio LC3/β-actin |
ratio Beclin/β-actin |
ratio LC3/β-actin |
|
0 | 0.67 | 1.18 | 0.81 | 1.35 |
10 | 0.76 | 0.86 | 0.63 | 1.59 |
25 | 0.37 | 1.05 | 0.82 | 2.49 |
50 | 0.41 | 2.55 | 1.39 | 2.28 |
100 | 0.25 | 3.47 | 1.10 | 4.57 |
Experimental Example 5. inhibitory effect on arthritis (
in vivo
).
To confirm the inhibitory effect of inventive extract on arthritis, the animal model test using by arthritis-induced rat animal model, was performed according to the previous known procedure in the art.
5-1. test procedure
In order to evaluate the efficacy of test sample on MIA(Monosodium iodoacetate)-induced osteoarthritis rat model, following test was performed at “Joint and Immune disease T2B center(chief MD. PARK, Sung-whan) located in “The Catholic University of Korea, Seoul ST. Mary’s Hospital, Seoul).
5-1-1. test protocol
5-1-1-1. animal model test procedure
(1)Test animals: Rats (Male Wista rats, Central Lab. Animal, Seoul, Korea, 7 to 8 weeks aged, 200 to 250g) were bred in the well-controlled breeding room in polysulfone cage (2-3 mice per cage) maintaining the temperature of 21 ± 2℃ and relative humidity of 50 ± 20% with the light cycle of day/dark (08:00 ~ 20:00) at the interval of 12 hours and acclimated to the surround environment
Test method: 3mg/kg of Monosodium iodoacetate (MIA, I2512, Sigma, Poole, UK) was dissolved in the injection saline to re reach to 60 mg/ml concentration on the day of the experiment (day 0). After dividing each groups, the experimental animals were placed in the anesthesia chamber and anesthetized with diethyl ether. 50 μL of MIA (3 mg/body) was injected into the right knee joint through the infrapatellar ligament using a 1cc syringe (26.5 gauge) in order to inducing osteoarthritis
(2)Test sample: the inventive extract prepared in Examples
(3)Positive control: Celecoxib® (Hanlim Pharmaceutical Company, Seoul, KOREA)
(4)Treatment route: oral administration 3days after inducing osteoarthritis (once a day)
(5)The establishment of test groups: See Table 8.
5-1-1-2.administration route and test period
(1)administration route
After inducing osteoarthritis with MIA, test sample treatment groups (G2, G3) were prepared by homogenizing the test samples with vehicle (saline) according to the prescribed dose and orally administrating 1 mL of test sample once a day.
The negative control group (G1) was treated with only vehicle and orally administrated once a day according to the sample schedule as the test substance administration.
The positive control group (G4) was homogenized to the vehicle according to the prescribed dose and orally administered once a day.
(2)test period
24 rats were divided into 4 groups per 6 rats, and the test samples and positive control substance were orally administered at a specified time.
Group | Drug administration | Dose | n |
G1 | MIA + vehicle (saline) | - | 6 |
G2 | MIA + WIN1001X (saline) | 100 ㎎/㎏ | 6 |
G3 | MIA + WIN1001X (saline) | 150 ㎎/㎏ | 6 |
G4 | MIA + Celecoxib (0.5 % CMC) | 30 ㎎/㎏ | 6 |
*negative control group(G1), test sample group(G2, G3), positive control group (G4), n=24 |
5-2. evaluation contents
5-2-1. determination of pain threshold
5-2-1-1. determination method of pain threshold
The pain threshold (nociceptive latency, threshold) was determined by using dynamic plantar aesthesiometer (Ugo Basile, 37400, Comerio, Italy), a device that gradually increases the force on the feet of Male Wistarats over a certain period of time, according to a von Freestyle evaluation method to measure pain thresholds ((Kwon JY et al., Sci Rep. 2018 Sep 14;8(1):13832)
Prior to measurement, the animal was placed in an acrylic box with a wire mesh bench and acclimated for five minutes.
The pain threshold was determined by measuring the weight showing paw withdrawal behavior which animal withdraws its feet by applying a slow force of 0 to 50 g over 10 seconds using metal filaments in the center of each animal's right hind foot.
To avoid tissue damage, the cut-off threshold was set to 50 g, and the measurement time was set to the day before the inducing day of osteoarthritis by the treatment of MIA to calculate base line value in the normal control group (G1), test sample groups (G2, G3) and positive control group (G4) and the same value was obtained in the base line (day3).
The same value was measured before the treatment of test samples (day3) and the value was measured once a week at a certain time by administering the test samples in the articular cavity of rats.
The test result of pain threshold was calculated according to (a) paw withdrawal latencies (sec) and (b) paw withdrawal threshold (g).
5-2-1-2. test result of determination of pain threshold
As the test result of the pain measurement, it has been confirmed that the test sample group treated with100 and 150 mg/kg of test samples showed potent inhibitory effect on pain comparing with negative control group treated with vehicle, dose-dependently manner. (Table 9-10)
MIA inducement | vehicle (s) | WIN1001X 100 ㎎/㎏ (s) |
WIN1001X 150 ㎎/㎏ (s) |
Celecoxib 30 ㎎/㎏ (s) |
Before MIA treatment | 16.8 ±0.8 | 16.2 ± 0.6 | 16.5 ± 0.6 | 16.0 ± 1.5 |
3 days after MIA treatment | 9.4 ± 1.2 | 8.0 ± 1.1 | 9.7 ± 1.1 | 9.2 ± 2.1 |
6 days after MIA treatment | 8.6 ± 1.0 | 11.0 ± 1.3 | 11.2 ± 1.3 | 9.9 ± 2.2 |
11 days after MIA treatment | 9.1 ± 1.4 | 12.4 ± 2.0 | 11.4 ± 1.8 | 10.2 ± 0.9 |
13 days after MIA treatment | 9.0 ± 1.6 | 11.4 ± 1.0 | 12.1 ± 1.5 | 10.4 ± 0.9 |
17 days after MIA treatment | 9.5 ± 0.4 | 11.1 ± 0.4 | 13.1 ± 1.4 | 10.9 ± 1.8 |
21 days after MIA treatment | 9.0 ± 1.6 | 12.8 ± 1.6 | 13.5 ± 1.6 | 11.9 ± 1.5 |
25 days after MIA treatment | 9.0 ± 0.5 | 13.3 ± 0.5 | 13.9 ± 2.0 | 11.6 ± 1.0 |
MIA inducement | vehicle (g) | WIN1001X 100 ㎎/㎏ (g) |
WIN1001X 150 ㎎/㎏ (g) |
Celecoxib 30 ㎎/㎏ (g) |
Before MIA treatment | 33.6 ± 1.7 | 32.5 ± 1.2 | 33.0 ± 1.2 | 33.8 ± 2.9 |
3 days after MIA treatment | 19.0 ± 2.4 | 16.2 ± 2.1 | 19.7 ± 2.2 | 18.6 ± 4.1 |
6 days after MIA treatment | 17.4 ± 1.8 | 22.3 ± 2.7 | 22.6 ± 2.5 | 19.9 ± 4.3 |
11 days after MIA treatment | 18.5 ± 2.8 | 25.0 ± 4.0 | 23.1 ± 3.6 | 20.7 ± 1.7 |
13 days after MIA treatment | 18.3 ± 3.2 | 23.0 ± 2.0 | 24.5 ± 2.9 | 21.0 ± 1.6 |
17 days after MIA treatment | 19.2 ± 0.8 | 22.5 ± 0.9 | 26.3 ± 2.7 | 22.0 ± 3.4 |
21 days after MIA treatment | 18.3 ± 3.1 | 25.7 ± 3.3 | 27.3 ± 3.2 | 24.0 ± 2.9 |
25 days after MIA treatment | 18.2 ± 1.1 | 26.7 ± 0.9 | 27.9 ± 4.0 | 23.4 ± 1.9 |
5-2-2. test on the determination of weight bearing
5-2-2-1. determination of weight bearing
In order to measure the change in weight (or weight distribution) between normal hind legs (left) and arthritis-induced hind legs (right), the load of both hind feet was determined using testing apparatus (Model 600, IITC, USA) according to the method described in the literature (Kwon JY et al., Sci Rep. 2018 Sep 14;8(1):13832).
The test animal shall be correctly located in the holder and fixed so that both feet can be stepped symmetrically, since the load can be varied depending on the position of the animal's foot. The particular person performs the fixation to reduce the possible error to the maximum extent.
When each animal was correctly located in the holder, the machine was operated and the weight bearing of each group was measured twice for 5 seconds per measurement.
The mean value for each test result was digitized into the weight bearing(g) of each foot.
After obtaining a baseline value before MIA treatment (Day 0), the weight bearing in the normal control group (G1), test sample groups (G2, G3) and positive control group (G4), was determined at a certain time twice a week, 3 days before the treatment of test sample (day 3).
The test results of the weight load measurement were transformed into each weight bearing ratio according to below Math 1.
5-2-2-2. test result of weight bearing
As can be seen in Table 11, it has been confirmed that the test sample group potently improved the balance capability of both limbs comparing with the negative control group.
MIA inducement | vehicle (%) | WIN1001X 100 ㎎/㎏ (%) |
WIN1001X 150 ㎎/㎏ (%) |
Celecoxib 30 ㎎/㎏ (%) |
Before MIA inducement | 50.6 ± 1.2 | 49.6 ± 0.8 | 50.3 ± 1.3 | 49.3 ± 0.9 |
3 days after MIA inducement | 36.9 ± 1.8 | 37.9 ± 2.7 | 35.2 ± 2.4 | 34.7 ± 4.0 |
6 days after MIA inducement | 35.5 ± 1.5 | 38.3 ± 1.7 | 38.6 ± 1.3 | 38.5 ± 1.8 |
11 days after MIA inducement | 33.8 ± 2.9 | 41.0 ± 2.5 | 39.1 ± 3.0 | 40.0 ± 3.2 |
13 days after MIA inducement | 32.3 ± 2.8 | 43.6 ± 1.4 | 44.2 ± 1.5 | 44.5 ± 0.7 |
17 days after MIA inducement | 32.2 ± 1.7 | 42.9 ± 1.2 | 42.5 ± 1.3 | 43.9 ± 1.3 |
21 days after MIA inducement | 31.6 ± 2.0 | 40.6 ± 1.2 | 41.0 ± 1.5 | 39.7 ± 3.7 |
25 days after MIA inducement | 33.3 ± 1.8 | 42.3 ± 2.3 | 42.0 ± 1.4 | 41.7 ± 1.9 |
5-2-3. Inhibitory test on bone injury (histological analysis, Micro-CT)
In order to determine the inhibitory effect of inventive extract on bone injury, following histological analysis was performed according to the known method disclosed in the literature (Kwon JY et al., Sci. Rep. 2018 Sep 14;8(1):13832 ).
5-2-3-1. inhibitory effect on bone injury caused by osteoarthritis (histological analysis, Micro-CT)
In order to determine the inhibitory effect of inventive extract on bone injury caused by osteoarthritis, the MIA and test sample were administered to the right knee joint of sacrificed rats (male Wistar rats) and fixed with formalin.
The degree of bone injury around femur area of the knee joint in each group consisting of 3 rats, was determined using X-ray source (70kV, 142uA, AI 0.5mm filter, rotation step 0.6 °) and animal scanner (SKYSCAN1272 ex-vivo micro-CT, Bruker micro CT, Belgium) to perform micro-CT photography at 15 μm of Pixel resolution including section formation (NRecon), section rotation (Data Viewer), data Analysis (CTAN), Volume rendering generation (CTVox), and surface rendering (CTAN+CTVol).
5-2-3-2. test result of histological analysis (Micro-CT)
As can be seen in Table 12, it has been confirmed that the test sample group treated with100 and 150 mg/kg of test samples showed potent inhibitory effect on bone injury comparing with negative control group (vehicle group) through the test result of Micro-CT photography.
vehicle | WIN1001X 100 ㎎/㎏ |
WIN1001X 150 ㎎/㎏ |
Celecoxib 30 ㎎/㎏ |
|
Bone surface (%) | 45.7 ± 8.9 | 59.3 ± 2.9 | 56.7 ± 2.5 | 57.5 ± 0.5 |
5-2-4. Inhibitory test on bone injury (histopathological analysis)
In order to determine the inhibitory effect of inventive extract on bone injury, following histopathological analysis was performed according to the known method disclosed in the literature (Kwon JY et al., Sci Rep. 2018 Sep. 14;8(1):13832 ).
5-2-3-1. inhibitory effect on bone injury caused by osteoarthritis (histopathological analysis)
In order to determine the inhibitory effect of inventive extract on bone injury caused by osteoarthritis, the MIA and test sample were administered to the right knee joint of sacrificed rats (male Wistar rats) and the right knee joint of the rat fixed with formalin was sliced to stain with Safranin O.
The histopathological analysis was performed by photographing the femur area of knee joint (x200 fold). The test result was calculated and evaluated according to the known methods including Total Mankin score (Bulstra SK et al., 1989, Clin Orthop Relat Res: 294-302.) and OARSI score (Pritzker K.P.H. et al., 2006, Osteoarthritis Cartilage 14:13-29).
5-2-3-2. test result of histopathological analysis
As can be seen in Table 13 (total Mankin score method) and Table 14 (OARSI score), it has been confirmed that the test sample group treated with100 and 150 mg/kg of test samples showed potent inhibitory effect on bone injury comparing with negative control group (vehicle group) through the test result of histopathological analysis
vehicle | WIN1001X 100 ㎎/㎏ |
WIN1001X 150 ㎎/㎏ |
Celecoxib 30 ㎎/㎏ |
|
Total Mankin score | 7.5 ± 0.8 | 2.8 ± 0.6 | 4.7 ± 0.7 | 4 ±0.6 |
vehicle | WIN1001X 100 ㎎/㎏ |
WIN1001X 150 ㎎/㎏ |
Celecoxib 30 ㎎/㎏ |
|
OARSI score | 4 ± 0.4 | 1.9 ± 0.4 | 2.0 ± 0.2 | 2.4 ± 0.4 |
Statistics analysis
All data were expressed in mean and standard deviation (mean±SD), and statistical significance verification was determined to be significant (P < 0.05 ), using two-way ANOVA, one-way ANOVA, in GraphPad PRISM Version 5.0 (USA) analysis program.
Hereinafter, the formulating methods and kinds of excipients will be described, but the present invention is not limited to them. The representative preparation examples were described as follows.
Preparation of injection
WIN-1001X extract: 100mg
Sodium metabisulfite : 3.0mg
Methyl paraben : 0.8mg
Propyl paraben : 0.1mg
Distilled water for injection : optimum amount
Injection preparation was prepared by dissolving active component, controlling pH to about 7.5 and then filling all the components in 2㎖ ample and sterilizing by conventional injection preparation method.
Preparation of powder
WIN-1002X extract : 500mg
Corn Starch : 100mg
Lactose : 100mg
Talc : 10mg
Powder preparation was prepared by mixing above components and filling sealed package.
Preparation of tablet
WIN-1003X extract 200mg
Corn Starch 100mg
Lactose 100mg
Magnesium stearate optimum amount
Tablet preparation was prepared by mixing above components and entabletting.
Preparation of capsule
WIN-1004X extract : 100mg
Lactose : 50mg
Corn starch : 50mg
Talc : 2mg
Magnesium stearate optimum amount
Tablet preparation was prepared by mixing above components and filling gelatin capsule by conventional gelatin preparation method.
Preparation of liquid
WIN-1005X extract : 1000mg
Sugar : 20g
Polysaccharide : 20g
Lemon flavor : 20g
Liquid preparation was prepared by dissolving active component, and then filling all the components in 1000㎖ ample and sterilizing by conventional liquid preparation method.
Preparation of health food
WIN-1001X extract : 1000mg
Vitamin mixture : optimum amount
Vitamin A acetate : 70g
Vitamin E : 1.0mg
Vitamin B10 : 13mg
Vitamin B2 : 0.15mg
Vitamin B6 : 0.5mg
Vitamin B1 : 20.2g
Vitamin C : 10mg
Biotin : 10g
Amide nicotinic acid : 1.7mg
Folic acid : 50g
Calcium pantothenic acid : 0.5mg
Mineral mixture : optimum amount
Ferrous sulfate : 1.75mg
Zinc oxide : 0.82mg
Magnesium carbonate : 25.3mg
Monopotassium phosphate : 15mg
Dicalcium phosphate : 55mg
Potassium citrate : 90mg
Calcium carbonate : 100mg
Magnesium chloride : 24.8mg
The above mentioned vitamin and mineral mixture may be varied in many ways. Such variations are not to be regarded as a departure from the spirit and scope of the present invention.
Preparation of health beverage
WIN-1002X extract : 1000mg
Citric acid : 1000mg
Oligosaccharide : 100g
Apricot concentration : 2g
Taurine : 1g
Distilled water : 900㎖
Health beverage preparation was prepared by dissolving active component, mixing, stirred at 85℃ for 1 hour, filtered and then filling all the components in 1000㎖ ample and sterilizing by conventional health beverage preparation method.
The invention being thus described, it will be obvious that the same may be varied in many ways. Such variations are not to be regarded as a departure from the spirit and scope of the present invention, and all such modifications as would be obvious to one skilled in the art are intended to be included within the scope of the following claims.
As described in the present invention, the present invention provides an oral pharmaceutical composition comprising a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix, the present inventors demonstrated that the anti-inflammatory/anti-arthritic effects of inventive combined composition is potent by accomplishing in vitro experiments such as the inhibitory test on the expression of cytokines involved in inflammation (RPLPO, TSLP, GM-CSF and IL-1beta) (Experimental Example 1); Cell viability test on HT-29 and THP-1 cell (in vitro Experimental Example 2); Anti-inflammatory activity in THP-1 cell (in vitro, Experimental Example 3); inhibitory effect on autophagy activity (in vitro, Experimental Example 4.) as well as in vivo experiments such as inhibitory effect on arthritis using by arthritis-induced rat animal model (in vivo, Experimental Example 5), therefore, it is confirmed that inventive combined extract is very useful in the alleviation or treatment of inflammatory disease and arthritis disease as a form of oral pharmaceutical composition.
Claims (9)
- An oral pharmaceutical composition comprising a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix, as an active ingredient for preventing and treating arthritis diseases.
- The oral pharmaceutical composition according to claim 1, wherein said “combined herb extract” is (a) combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix with the mixed ratio based on the dried weight of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix (w/w) ranging from 0.01 -100: 0.01 -100: 0.01 -100 weight part (w/w); or (b) the combination of each extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix with the mixed ratio based on the dried weight of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix (w/w) ranging from 0.01 -100: 0.01 -100: 0.01 -100 weight part (w/w).
- The oral pharmaceutical composition according to claim 1, wherein said extract is extracted with at least one solvent selected from water methanol, ethanol, propanol, butanol, acetone, ethyl acetate, chloroform, hexane, butyleneglycol, propyleneglycol or glycerin.
- The oral pharmaceutical composition according to claim 1, wherein said arthritis diseases is selected from rigid spondylitis, degenerative arthritis, osteoarthritis, rheumatoid arthritis, gout or frozen shoulder.
- A health functional food comprising a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix, as an active ingredient to prevent or improve arthritis diseases.
- The health functional food according to claim 4, wherein said health functional food is provided as powder, granule, tablet, capsule, pill, suspension, emulsion, syrup, tea bag, leached tea, or beverage type.
- A health supplement food comprising a combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix, as an active ingredient or as a main component to prevent or improve arthritis diseases.
- A method of treating or alleviating arthritis diseases in a mammal comprising orally administering to said mammal an effective amount of the combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix and pharmaceutically acceptable carrier thereof.
- A use of the combined herb extract of Longanae Arillus, Ligustici Tenuissimi Rhizoma and Polygalae radix for manufacture of oral preparation employed for treating or alleviating arthritis diseases in mammals including human as an active ingredient.
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR20200040950 | 2020-04-03 | ||
KR10-2020-0040950 | 2020-04-03 | ||
KR10-2021-0032876 | 2021-03-12 | ||
KR1020210032876A KR102374385B1 (en) | 2020-04-03 | 2021-03-12 | a composition comprising the extract of combined herbs comprising Longanae Arillus for the treatment or prevention of arthritis |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2021201503A1 true WO2021201503A1 (en) | 2021-10-07 |
Family
ID=77927543
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/KR2021/003729 WO2021201503A1 (en) | 2020-04-03 | 2021-03-25 | An oral pharmaceutical composition comprising an extract of combined herbs comprising longanae arillus for the treatment or alleviation of arthritis disease and the use thereof. |
Country Status (1)
Country | Link |
---|---|
WO (1) | WO2021201503A1 (en) |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20050007996A (en) * | 2003-07-14 | 2005-01-21 | 주식회사 엠디바이오알파 | Compositions for arthritis disease |
KR20140082502A (en) * | 2012-12-24 | 2014-07-02 | 원광대학교산학협력단 | Composition comprising tenuifoliside A for preventing or treating inflammatory diseases |
KR20150115638A (en) * | 2014-04-04 | 2015-10-14 | (주)내츄럴엔도텍 | Compositions Comprising Extracts of Longan Arillus as Active Ingredients for Preventing or Treating an Osteoporosis |
KR20160054681A (en) * | 2014-11-06 | 2016-05-17 | 주식회사 엘지생활건강 | Composition for promoting synthesis of hyaluronic acid comprising Polygala tenuifolia Willd extracts and the use thereof |
KR20190100880A (en) * | 2018-02-21 | 2019-08-29 | 경북대학교 산학협력단 | Composition for preventing, treating or improving obesity or obesity-related disease comprising extracts of Angelica tenuissima |
-
2021
- 2021-03-25 WO PCT/KR2021/003729 patent/WO2021201503A1/en active Application Filing
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20050007996A (en) * | 2003-07-14 | 2005-01-21 | 주식회사 엠디바이오알파 | Compositions for arthritis disease |
KR20140082502A (en) * | 2012-12-24 | 2014-07-02 | 원광대학교산학협력단 | Composition comprising tenuifoliside A for preventing or treating inflammatory diseases |
KR20150115638A (en) * | 2014-04-04 | 2015-10-14 | (주)내츄럴엔도텍 | Compositions Comprising Extracts of Longan Arillus as Active Ingredients for Preventing or Treating an Osteoporosis |
KR20160054681A (en) * | 2014-11-06 | 2016-05-17 | 주식회사 엘지생활건강 | Composition for promoting synthesis of hyaluronic acid comprising Polygala tenuifolia Willd extracts and the use thereof |
KR20190100880A (en) * | 2018-02-21 | 2019-08-29 | 경북대학교 산학협력단 | Composition for preventing, treating or improving obesity or obesity-related disease comprising extracts of Angelica tenuissima |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
WO2012074183A1 (en) | Pharmaceutical composition for preventing or treating inflammatory diseases comprising trachelospermi caulis extract and paeonia suffruticosa andrews extract, and method for preparing the same | |
WO2013176512A1 (en) | Panax spp. plant extract with increased content ratio of ginsenoside rg3, rg5, and rk1 produced by microwave irradiation, a method of preparing the panax spp. plant extract, and a composition including the panax spp. plant extract | |
WO2012124888A2 (en) | Composition comprising the extract of herbal combination for preventing or treating diabetic peripheral neuropathy | |
WO2018174448A1 (en) | Composition for treating and preventing climacteric syndrome containing combined herbal medicinal extract of white atractylis, mori fructus, chinese matrimony vine, longan, achyranthes, eucommia bark, and asparagus cochinchinensis merr. as active ingredient, and use of same | |
WO2010087577A2 (en) | Use of thymus capitatus extract, satureja hortensis extract, or carvacrol for treating metabolic diseases | |
WO2017183924A1 (en) | Composition for preventing or treating dry eye syndrome comprising maple leaf extract | |
WO2021201503A1 (en) | An oral pharmaceutical composition comprising an extract of combined herbs comprising longanae arillus for the treatment or alleviation of arthritis disease and the use thereof. | |
WO2012093787A2 (en) | Composition for activating gabaa benzodiazepine receptor and composition for anxiety alleviation, convulsion reduction, sedation and sleep induction and improvement containing phloroglucinol, phlorotannin or brown algae extract | |
WO2021201502A1 (en) | An oral pharmaceutical composition comprising an extract of combined herbs comprising longanae arillus for the treatment or alleviation of inflammatory disease and the use thereof. | |
WO2012081831A2 (en) | Composition comprising the extract of loranthus yadoriki sieb having monoamine oxidase-inhibiting activity | |
WO2015002430A1 (en) | Pharmaceutical composition for preventing or treating asthma comprising pistacia weinmannifolia j. poiss. ex franch extract or fraction thereof | |
WO2010143825A2 (en) | Anti-arthritic agent using cyathula officinalis | |
WO2020145619A1 (en) | Composition for allergy prevention, atopic dermatitis alleviation or skin regeneration, containing, as active ingredient, undecane or undecanal | |
WO2021256858A1 (en) | Composition for improving, preventing or treating non-alcoholic fatty liver disease and preparation method therefor | |
WO2020130478A1 (en) | Anti-allergy, atopic dermatitis alleviation or skin regeneration composition containing jasmone as active ingredient | |
WO2009104900A2 (en) | Composition comprising a flower extract of lonicera japonica thunb for preventing and treating arthritic diseases | |
WO2010090423A2 (en) | Composition comprising combined crude drug extract for preventing and treating hyperlipidemia and diabetic hyperlipidemia | |
WO2015105373A1 (en) | Composition for prevention or treatment of asthma, comprising e uonymus alatus extract or fraction thereof | |
WO2019039810A1 (en) | Composition comprising extract of lonicera coerulea for prevention or treatment of pain | |
WO2009104901A2 (en) | Composition comprising an extract of anemarrhena asphodeloides bunge for preventing and treating arthritic diseases | |
WO2014126269A1 (en) | Pharmaceutical composition for improving male sexual function and for treating infertility | |
WO2011081231A1 (en) | Composition and functional food using polygonum cuspidatum butanol extract or ethyl acetate extract for treating and preventing obesity | |
WO2020218733A2 (en) | Composition for alleviating, preventing or treating arthritis, containing stewartia pseudocamellia leaf extract and maclura tricuspidata leaf extract as active ingredients | |
WO2023128636A1 (en) | Composition containing brier tree root extract for alleviation, prevention, or treatment of obesity and metabolic diseases | |
WO2021182864A1 (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. |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 21780149 Country of ref document: EP Kind code of ref document: A1 |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
122 | Ep: pct application non-entry in european phase |
Ref document number: 21780149 Country of ref document: EP Kind code of ref document: A1 |