KR20210048208A - Composition for preventing or treating TNF-mediated diseases comprising tetrahydropapaverine and method for inhibiting TNF-activity with the same - Google Patents
Composition for preventing or treating TNF-mediated diseases comprising tetrahydropapaverine and method for inhibiting TNF-activity with the same Download PDFInfo
- Publication number
- KR20210048208A KR20210048208A KR1020190132187A KR20190132187A KR20210048208A KR 20210048208 A KR20210048208 A KR 20210048208A KR 1020190132187 A KR1020190132187 A KR 1020190132187A KR 20190132187 A KR20190132187 A KR 20190132187A KR 20210048208 A KR20210048208 A KR 20210048208A
- Authority
- KR
- South Korea
- Prior art keywords
- tnf
- disease
- diseases
- treating
- preventing
- Prior art date
Links
- 201000010099 disease Diseases 0.000 title claims abstract description 31
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title claims abstract description 31
- 239000000203 mixture Substances 0.000 title claims abstract description 26
- 230000000694 effects Effects 0.000 title claims abstract description 20
- 238000000034 method Methods 0.000 title claims abstract description 19
- 230000002401 inhibitory effect Effects 0.000 title claims abstract description 16
- 102100040247 Tumor necrosis factor Human genes 0.000 title claims abstract 19
- YXWQTVWJNHKSCC-UHFFFAOYSA-N 1-[(3,4-dimethoxyphenyl)methyl]-6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline Chemical compound C1=C(OC)C(OC)=CC=C1CC1C2=CC(OC)=C(OC)C=C2CCN1 YXWQTVWJNHKSCC-UHFFFAOYSA-N 0.000 title claims description 49
- 230000001404 mediated effect Effects 0.000 title 1
- 210000000068 Th17 cell Anatomy 0.000 claims abstract description 20
- 230000004069 differentiation Effects 0.000 claims abstract description 15
- 239000004480 active ingredient Substances 0.000 claims abstract description 13
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 13
- 235000013402 health food Nutrition 0.000 claims abstract description 10
- 210000002437 synoviocyte Anatomy 0.000 claims abstract description 9
- 230000035755 proliferation Effects 0.000 claims abstract description 7
- 210000000988 bone and bone Anatomy 0.000 claims abstract description 5
- 206010039073 rheumatoid arthritis Diseases 0.000 claims description 16
- 230000014509 gene expression Effects 0.000 claims description 14
- 206010003246 arthritis Diseases 0.000 claims description 11
- 201000004624 Dermatitis Diseases 0.000 claims description 8
- 238000000338 in vitro Methods 0.000 claims description 8
- 108090001005 Interleukin-6 Proteins 0.000 claims description 7
- 201000004681 Psoriasis Diseases 0.000 claims description 6
- 241001465754 Metazoa Species 0.000 claims description 5
- 208000023275 Autoimmune disease Diseases 0.000 claims description 4
- 206010015150 Erythema Diseases 0.000 claims description 4
- 206010060862 Prostate cancer Diseases 0.000 claims description 4
- 208000000236 Prostatic Neoplasms Diseases 0.000 claims description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- 231100000321 erythema Toxicity 0.000 claims description 4
- 201000008383 nephritis Diseases 0.000 claims description 4
- 208000017520 skin disease Diseases 0.000 claims description 4
- 208000002249 Diabetes Complications Diseases 0.000 claims description 3
- 208000003456 Juvenile Arthritis Diseases 0.000 claims description 3
- 206010059176 Juvenile idiopathic arthritis Diseases 0.000 claims description 3
- 230000001363 autoimmune Effects 0.000 claims description 3
- 206010012601 diabetes mellitus Diseases 0.000 claims description 3
- 201000002215 juvenile rheumatoid arthritis Diseases 0.000 claims description 3
- 231100000331 toxic Toxicity 0.000 claims description 3
- 230000002588 toxic effect Effects 0.000 claims description 3
- 206010000748 Acute febrile neutrophilic dermatosis Diseases 0.000 claims description 2
- 201000004384 Alopecia Diseases 0.000 claims description 2
- 208000024827 Alzheimer disease Diseases 0.000 claims description 2
- 206010002556 Ankylosing Spondylitis Diseases 0.000 claims description 2
- 208000019901 Anxiety disease Diseases 0.000 claims description 2
- 206010003210 Arteriosclerosis Diseases 0.000 claims description 2
- 208000025705 Axial Spondyloarthritis Diseases 0.000 claims description 2
- 208000035143 Bacterial infection Diseases 0.000 claims description 2
- 208000009137 Behcet syndrome Diseases 0.000 claims description 2
- 208000020925 Bipolar disease Diseases 0.000 claims description 2
- 206010005003 Bladder cancer Diseases 0.000 claims description 2
- 206010006187 Breast cancer Diseases 0.000 claims description 2
- 208000026310 Breast neoplasm Diseases 0.000 claims description 2
- 206010006895 Cachexia Diseases 0.000 claims description 2
- 208000024172 Cardiovascular disease Diseases 0.000 claims description 2
- 206010063094 Cerebral malaria Diseases 0.000 claims description 2
- 208000006545 Chronic Obstructive Pulmonary Disease Diseases 0.000 claims description 2
- 206010009900 Colitis ulcerative Diseases 0.000 claims description 2
- 206010009944 Colon cancer Diseases 0.000 claims description 2
- 208000035473 Communicable disease Diseases 0.000 claims description 2
- 208000011231 Crohn disease Diseases 0.000 claims description 2
- 208000006313 Delayed Hypersensitivity Diseases 0.000 claims description 2
- 206010012289 Dementia Diseases 0.000 claims description 2
- 206010012438 Dermatitis atopic Diseases 0.000 claims description 2
- 206010012442 Dermatitis contact Diseases 0.000 claims description 2
- 206010048768 Dermatosis Diseases 0.000 claims description 2
- 206010012688 Diabetic retinal oedema Diseases 0.000 claims description 2
- 208000004232 Enteritis Diseases 0.000 claims description 2
- 206010014954 Eosinophilic fasciitis Diseases 0.000 claims description 2
- 206010059284 Epidermal necrosis Diseases 0.000 claims description 2
- 206010061857 Fat necrosis Diseases 0.000 claims description 2
- 206010017533 Fungal infection Diseases 0.000 claims description 2
- 206010017711 Gangrene Diseases 0.000 claims description 2
- 208000010412 Glaucoma Diseases 0.000 claims description 2
- 206010018338 Glioma Diseases 0.000 claims description 2
- 208000009329 Graft vs Host Disease Diseases 0.000 claims description 2
- 206010018691 Granuloma Diseases 0.000 claims description 2
- 206010072579 Granulomatosis with polyangiitis Diseases 0.000 claims description 2
- 206010019280 Heart failures Diseases 0.000 claims description 2
- 208000032759 Hemolytic-Uremic Syndrome Diseases 0.000 claims description 2
- 208000031226 Hyperlipidaemia Diseases 0.000 claims description 2
- 206010020751 Hypersensitivity Diseases 0.000 claims description 2
- 208000022559 Inflammatory bowel disease Diseases 0.000 claims description 2
- 208000011200 Kawasaki disease Diseases 0.000 claims description 2
- 208000008839 Kidney Neoplasms Diseases 0.000 claims description 2
- 206010067125 Liver injury Diseases 0.000 claims description 2
- 206010025323 Lymphomas Diseases 0.000 claims description 2
- 208000034578 Multiple myelomas Diseases 0.000 claims description 2
- 208000031888 Mycoses Diseases 0.000 claims description 2
- 201000003793 Myelodysplastic syndrome Diseases 0.000 claims description 2
- 206010028980 Neoplasm Diseases 0.000 claims description 2
- 208000012902 Nervous system disease Diseases 0.000 claims description 2
- 208000025966 Neurological disease Diseases 0.000 claims description 2
- 208000008589 Obesity Diseases 0.000 claims description 2
- 206010033078 Otitis media Diseases 0.000 claims description 2
- 206010033128 Ovarian cancer Diseases 0.000 claims description 2
- 206010061535 Ovarian neoplasm Diseases 0.000 claims description 2
- 208000002193 Pain Diseases 0.000 claims description 2
- 206010061902 Pancreatic neoplasm Diseases 0.000 claims description 2
- 206010033645 Pancreatitis Diseases 0.000 claims description 2
- 208000018737 Parkinson disease Diseases 0.000 claims description 2
- 241000721454 Pemphigus Species 0.000 claims description 2
- 206010035226 Plasma cell myeloma Diseases 0.000 claims description 2
- 206010035664 Pneumonia Diseases 0.000 claims description 2
- 201000001263 Psoriatic Arthritis Diseases 0.000 claims description 2
- 208000036824 Psoriatic arthropathy Diseases 0.000 claims description 2
- 208000006311 Pyoderma Diseases 0.000 claims description 2
- 206010038389 Renal cancer Diseases 0.000 claims description 2
- 206010063837 Reperfusion injury Diseases 0.000 claims description 2
- 206010040047 Sepsis Diseases 0.000 claims description 2
- 206010040070 Septic Shock Diseases 0.000 claims description 2
- 201000001880 Sexual dysfunction Diseases 0.000 claims description 2
- 208000021386 Sjogren Syndrome Diseases 0.000 claims description 2
- 208000000453 Skin Neoplasms Diseases 0.000 claims description 2
- 208000004078 Snake Bites Diseases 0.000 claims description 2
- 201000002661 Spondylitis Diseases 0.000 claims description 2
- 208000010443 Steatitis Diseases 0.000 claims description 2
- 208000006011 Stroke Diseases 0.000 claims description 2
- 208000010265 Sweet syndrome Diseases 0.000 claims description 2
- 201000009594 Systemic Scleroderma Diseases 0.000 claims description 2
- 206010042953 Systemic sclerosis Diseases 0.000 claims description 2
- 208000024799 Thyroid disease Diseases 0.000 claims description 2
- 201000006704 Ulcerative Colitis Diseases 0.000 claims description 2
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 claims description 2
- 206010046851 Uveitis Diseases 0.000 claims description 2
- 206010047115 Vasculitis Diseases 0.000 claims description 2
- 208000036142 Viral infection Diseases 0.000 claims description 2
- 230000007815 allergy Effects 0.000 claims description 2
- 231100000360 alopecia Toxicity 0.000 claims description 2
- 206010002026 amyotrophic lateral sclerosis Diseases 0.000 claims description 2
- 208000011775 arteriosclerosis disease Diseases 0.000 claims description 2
- 208000006673 asthma Diseases 0.000 claims description 2
- 201000008937 atopic dermatitis Diseases 0.000 claims description 2
- 208000022362 bacterial infectious disease Diseases 0.000 claims description 2
- 201000011510 cancer Diseases 0.000 claims description 2
- 208000015114 central nervous system disease Diseases 0.000 claims description 2
- 208000019069 chronic childhood arthritis Diseases 0.000 claims description 2
- 208000029742 colonic neoplasm Diseases 0.000 claims description 2
- 208000010247 contact dermatitis Diseases 0.000 claims description 2
- 230000006378 damage Effects 0.000 claims description 2
- 201000011190 diabetic macular edema Diseases 0.000 claims description 2
- 230000003325 follicular Effects 0.000 claims description 2
- 230000004927 fusion Effects 0.000 claims description 2
- 208000024908 graft versus host disease Diseases 0.000 claims description 2
- 208000006454 hepatitis Diseases 0.000 claims description 2
- 231100000283 hepatitis Toxicity 0.000 claims description 2
- 208000026278 immune system disease Diseases 0.000 claims description 2
- 208000027866 inflammatory disease Diseases 0.000 claims description 2
- 230000000366 juvenile effect Effects 0.000 claims description 2
- 201000010982 kidney cancer Diseases 0.000 claims description 2
- 208000017169 kidney disease Diseases 0.000 claims description 2
- 208000032839 leukemia Diseases 0.000 claims description 2
- 208000019423 liver disease Diseases 0.000 claims description 2
- 206010025135 lupus erythematosus Diseases 0.000 claims description 2
- 208000002780 macular degeneration Diseases 0.000 claims description 2
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 claims description 2
- 208000030159 metabolic disease Diseases 0.000 claims description 2
- 208000001725 mucocutaneous lymph node syndrome Diseases 0.000 claims description 2
- 201000006417 multiple sclerosis Diseases 0.000 claims description 2
- 208000010125 myocardial infarction Diseases 0.000 claims description 2
- 208000031225 myocardial ischemia Diseases 0.000 claims description 2
- 230000003448 neutrophilic effect Effects 0.000 claims description 2
- 235000020824 obesity Nutrition 0.000 claims description 2
- 210000000056 organ Anatomy 0.000 claims description 2
- 201000008482 osteoarthritis Diseases 0.000 claims description 2
- 201000008968 osteosarcoma Diseases 0.000 claims description 2
- 201000002528 pancreatic cancer Diseases 0.000 claims description 2
- 208000008443 pancreatic carcinoma Diseases 0.000 claims description 2
- 208000005987 polymyositis Diseases 0.000 claims description 2
- 201000011461 pre-eclampsia Diseases 0.000 claims description 2
- 208000020016 psychiatric disease Diseases 0.000 claims description 2
- 201000000306 sarcoidosis Diseases 0.000 claims description 2
- 230000036303 septic shock Effects 0.000 claims description 2
- 231100000872 sexual dysfunction Toxicity 0.000 claims description 2
- 201000000849 skin cancer Diseases 0.000 claims description 2
- 201000000596 systemic lupus erythematosus Diseases 0.000 claims description 2
- 206010043778 thyroiditis Diseases 0.000 claims description 2
- 201000008827 tuberculosis Diseases 0.000 claims description 2
- 201000005112 urinary bladder cancer Diseases 0.000 claims description 2
- 230000009385 viral infection Effects 0.000 claims description 2
- 206010012655 Diabetic complications Diseases 0.000 claims 1
- 206010010164 Hypertension complications Diseases 0.000 claims 1
- 206010028741 Nasal inflammation Diseases 0.000 claims 1
- 208000007048 Polymyalgia Rheumatica Diseases 0.000 claims 1
- 208000034189 Sclerosis Diseases 0.000 claims 1
- 125000004122 cyclic group Chemical group 0.000 claims 1
- 231100000234 hepatic damage Toxicity 0.000 claims 1
- 201000008298 histiocytosis Diseases 0.000 claims 1
- 208000015181 infectious disease Diseases 0.000 claims 1
- 210000003734 kidney Anatomy 0.000 claims 1
- 230000008818 liver damage Effects 0.000 claims 1
- 230000035900 sweating Effects 0.000 claims 1
- 208000020408 systemic-onset juvenile idiopathic arthritis Diseases 0.000 claims 1
- 208000009386 Experimental Arthritis Diseases 0.000 abstract description 14
- 102000004127 Cytokines Human genes 0.000 abstract description 13
- 108090000695 Cytokines Proteins 0.000 abstract description 13
- 230000002757 inflammatory effect Effects 0.000 abstract description 7
- 210000000845 cartilage Anatomy 0.000 abstract description 4
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 60
- 102000003390 tumor necrosis factor Human genes 0.000 description 54
- 210000004027 cell Anatomy 0.000 description 30
- 241000699666 Mus <mouse, genus> Species 0.000 description 19
- 102000013691 Interleukin-17 Human genes 0.000 description 14
- 108050003558 Interleukin-17 Proteins 0.000 description 14
- 239000002953 phosphate buffered saline Substances 0.000 description 14
- 238000005406 washing Methods 0.000 description 14
- 230000027455 binding Effects 0.000 description 10
- -1 small molecule compound Chemical class 0.000 description 9
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 8
- 238000002360 preparation method Methods 0.000 description 8
- 102000004889 Interleukin-6 Human genes 0.000 description 7
- 241000699670 Mus sp. Species 0.000 description 7
- 238000011529 RT qPCR Methods 0.000 description 7
- 230000001419 dependent effect Effects 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 6
- 238000000684 flow cytometry Methods 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 239000000546 pharmaceutical excipient Substances 0.000 description 6
- 230000028327 secretion Effects 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 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 5
- 238000002835 absorbance Methods 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 230000024245 cell differentiation Effects 0.000 description 5
- 239000012228 culture supernatant Substances 0.000 description 5
- 210000002683 foot Anatomy 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 108020003175 receptors Proteins 0.000 description 5
- 102000005962 receptors Human genes 0.000 description 5
- 230000002829 reductive effect Effects 0.000 description 5
- 238000002965 ELISA Methods 0.000 description 4
- 108010008165 Etanercept Proteins 0.000 description 4
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 4
- 210000001744 T-lymphocyte Anatomy 0.000 description 4
- RJURFGZVJUQBHK-UHFFFAOYSA-N actinomycin D Natural products CC1OC(=O)C(C(C)C)N(C)C(=O)CN(C)C(=O)C2CCCN2C(=O)C(C(C)C)NC(=O)C1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=CC=C3C(=O)NC4C(=O)NC(C(N5CCCC5C(=O)N(C)CC(=O)N(C)C(C(C)C)C(=O)OC4C)=O)C(C)C)=C3N=C21 RJURFGZVJUQBHK-UHFFFAOYSA-N 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 4
- 108020004999 messenger RNA Proteins 0.000 description 4
- 230000000770 proinflammatory effect Effects 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 210000004988 splenocyte Anatomy 0.000 description 4
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 4
- 239000003981 vehicle Substances 0.000 description 4
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 244000299461 Theobroma cacao Species 0.000 description 3
- 230000000903 blocking effect Effects 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 230000004663 cell proliferation Effects 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
- 230000003013 cytotoxicity Effects 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 210000002950 fibroblast Anatomy 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000013373 food additive Nutrition 0.000 description 3
- 239000002778 food additive Substances 0.000 description 3
- 235000013355 food flavoring agent Nutrition 0.000 description 3
- 230000003053 immunization Effects 0.000 description 3
- 238000002649 immunization Methods 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 230000003472 neutralizing effect Effects 0.000 description 3
- 239000003755 preservative agent Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 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
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 108010092160 Dactinomycin Proteins 0.000 description 2
- 238000008157 ELISA kit Methods 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 101000998146 Homo sapiens Interleukin-17A Proteins 0.000 description 2
- 238000012404 In vitro experiment Methods 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 102000051628 Interleukin-1 receptor antagonist Human genes 0.000 description 2
- 108700021006 Interleukin-1 receptor antagonist Proteins 0.000 description 2
- 108090001007 Interleukin-8 Proteins 0.000 description 2
- 102000004890 Interleukin-8 Human genes 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- 102000001691 Member 3 Group F Nuclear Receptor Subfamily 1 Human genes 0.000 description 2
- 108010029279 Member 3 Group F Nuclear Receptor Subfamily 1 Proteins 0.000 description 2
- 101000998145 Mus musculus Interleukin-17A Proteins 0.000 description 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
- 229930182555 Penicillin Natural products 0.000 description 2
- 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 2
- 229920001213 Polysorbate 20 Polymers 0.000 description 2
- 108091008778 RORγ2 Proteins 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 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 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 101710187830 Tumor necrosis factor receptor superfamily member 1B Proteins 0.000 description 2
- 102100033733 Tumor necrosis factor receptor superfamily member 1B Human genes 0.000 description 2
- RJURFGZVJUQBHK-IIXSONLDSA-N actinomycin D Chemical compound C[C@H]1OC(=O)[C@H](C(C)C)N(C)C(=O)CN(C)C(=O)[C@@H]2CCCN2C(=O)[C@@H](C(C)C)NC(=O)[C@H]1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=CC=C3C(=O)N[C@@H]4C(=O)N[C@@H](C(N5CCC[C@H]5C(=O)N(C)CC(=O)N(C)[C@@H](C(C)C)C(=O)O[C@@H]4C)=O)C(C)C)=C3N=C21 RJURFGZVJUQBHK-IIXSONLDSA-N 0.000 description 2
- 230000004913 activation Effects 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
- 238000010171 animal model Methods 0.000 description 2
- 239000003125 aqueous solvent Substances 0.000 description 2
- 108010030694 avidin-horseradish peroxidase complex Proteins 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- 235000013361 beverage Nutrition 0.000 description 2
- 239000011230 binding agent Substances 0.000 description 2
- 229940125385 biologic drug Drugs 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 235000010980 cellulose Nutrition 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 235000019219 chocolate Nutrition 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 229960000640 dactinomycin Drugs 0.000 description 2
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 229940073621 enbrel Drugs 0.000 description 2
- UVCJGUGAGLDPAA-UHFFFAOYSA-N ensulizole Chemical compound N1C2=CC(S(=O)(=O)O)=CC=C2N=C1C1=CC=CC=C1 UVCJGUGAGLDPAA-UHFFFAOYSA-N 0.000 description 2
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 2
- 229960000403 etanercept Drugs 0.000 description 2
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 235000015203 fruit juice Nutrition 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 210000000548 hind-foot Anatomy 0.000 description 2
- 102000053162 human IL17A Human genes 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 210000005067 joint tissue Anatomy 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 239000012139 lysis buffer Substances 0.000 description 2
- 210000002540 macrophage Anatomy 0.000 description 2
- 235000019359 magnesium stearate Nutrition 0.000 description 2
- 238000002826 magnetic-activated cell sorting Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 239000011325 microbead Substances 0.000 description 2
- 210000001616 monocyte Anatomy 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- 229940049954 penicillin Drugs 0.000 description 2
- 230000008823 permeabilization Effects 0.000 description 2
- 239000000049 pigment Substances 0.000 description 2
- 229920009537 polybutylene succinate adipate Polymers 0.000 description 2
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 2
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 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 2
- 239000007787 solid Substances 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000012089 stop solution Substances 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000000829 suppository Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- 239000000454 talc Substances 0.000 description 2
- 229910052623 talc Inorganic materials 0.000 description 2
- 235000012222 talc Nutrition 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 239000002562 thickening agent Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 239000000080 wetting agent Substances 0.000 description 2
- HMLGSIZOMSVISS-ONJSNURVSA-N (7r)-7-[[(2z)-2-(2-amino-1,3-thiazol-4-yl)-2-(2,2-dimethylpropanoyloxymethoxyimino)acetyl]amino]-3-ethenyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid Chemical compound N([C@@H]1C(N2C(=C(C=C)CSC21)C(O)=O)=O)C(=O)\C(=N/OCOC(=O)C(C)(C)C)C1=CSC(N)=N1 HMLGSIZOMSVISS-ONJSNURVSA-N 0.000 description 1
- WBYWAXJHAXSJNI-VOTSOKGWSA-M .beta-Phenylacrylic acid Natural products [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 description 1
- AZKSAVLVSZKNRD-UHFFFAOYSA-M 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Chemical compound [Br-].S1C(C)=C(C)N=C1[N+]1=NC(C=2C=CC=CC=2)=NN1C1=CC=CC=C1 AZKSAVLVSZKNRD-UHFFFAOYSA-M 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-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
- 244000215068 Acacia senegal Species 0.000 description 1
- 235000006491 Acacia senegal Nutrition 0.000 description 1
- 102000010565 Apoptosis Regulatory Proteins Human genes 0.000 description 1
- 108010063104 Apoptosis Regulatory Proteins Proteins 0.000 description 1
- 206010051728 Bone erosion Diseases 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 101710155857 C-C motif chemokine 2 Proteins 0.000 description 1
- 102100021943 C-C motif chemokine 2 Human genes 0.000 description 1
- 102100032367 C-C motif chemokine 5 Human genes 0.000 description 1
- 238000011740 C57BL/6 mouse Methods 0.000 description 1
- 206010007710 Cartilage injury Diseases 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
- 108010055166 Chemokine CCL5 Proteins 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- 102000019034 Chemokines Human genes 0.000 description 1
- WBYWAXJHAXSJNI-SREVYHEPSA-N Cinnamic acid Chemical compound OC(=O)\C=C/C1=CC=CC=C1 WBYWAXJHAXSJNI-SREVYHEPSA-N 0.000 description 1
- 102000000503 Collagen Type II Human genes 0.000 description 1
- 108010041390 Collagen Type II Proteins 0.000 description 1
- 241000699800 Cricetinae Species 0.000 description 1
- 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 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
- 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 1
- 238000011763 DBA/1J (JAX™ mouse strain) Methods 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- LVGKNOAMLMIIKO-UHFFFAOYSA-N Elaidinsaeure-aethylester Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC LVGKNOAMLMIIKO-UHFFFAOYSA-N 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 239000004386 Erythritol Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- 208000007882 Gastritis Diseases 0.000 description 1
- 208000032612 Glial tumor Diseases 0.000 description 1
- 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 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 1
- 229920002907 Guar gum Polymers 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 101100005713 Homo sapiens CD4 gene Proteins 0.000 description 1
- 101000883515 Homo sapiens Chitinase-3-like protein 1 Proteins 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 102000000589 Interleukin-1 Human genes 0.000 description 1
- 102000019223 Interleukin-1 receptor Human genes 0.000 description 1
- 108050006617 Interleukin-1 receptor Proteins 0.000 description 1
- 102000010781 Interleukin-6 Receptors Human genes 0.000 description 1
- 108010038501 Interleukin-6 Receptors Proteins 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- 229930182816 L-glutamine Natural products 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 101710151805 Mitochondrial intermediate peptidase 1 Proteins 0.000 description 1
- 229930191564 Monensin Natural products 0.000 description 1
- GAOZTHIDHYLHMS-UHFFFAOYSA-N Monensin A Natural products O1C(CC)(C2C(CC(O2)C2C(CC(C)C(O)(CO)O2)C)C)CCC1C(O1)(C)CCC21CC(O)C(C)C(C(C)C(OC)C(C)C(O)=O)O2 GAOZTHIDHYLHMS-UHFFFAOYSA-N 0.000 description 1
- 101001033286 Mus musculus Interleukin-1 beta Proteins 0.000 description 1
- 101001076414 Mus musculus Interleukin-6 Proteins 0.000 description 1
- 101100046526 Mus musculus Tnf gene Proteins 0.000 description 1
- 208000000112 Myalgia Diseases 0.000 description 1
- 102000003945 NF-kappa B Human genes 0.000 description 1
- 108010057466 NF-kappa B Proteins 0.000 description 1
- 206010029350 Neurotoxicity Diseases 0.000 description 1
- 229920002230 Pectic acid Polymers 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 206010061481 Renal injury Diseases 0.000 description 1
- 206010039710 Scleroderma Diseases 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 244000269722 Thea sinensis 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
- 206010044221 Toxic encephalopathy Diseases 0.000 description 1
- 101710187743 Tumor necrosis factor receptor superfamily member 1A Proteins 0.000 description 1
- 102100033732 Tumor necrosis factor receptor superfamily member 1A Human genes 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 229940119059 actemra Drugs 0.000 description 1
- 229960002964 adalimumab Drugs 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 150000001298 alcohols Chemical class 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
- 239000003513 alkali Substances 0.000 description 1
- 229960004238 anakinra Drugs 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 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 1
- 238000013357 binding ELISA Methods 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 235000008429 bread Nutrition 0.000 description 1
- 235000014121 butter Nutrition 0.000 description 1
- 235000001046 cacaotero Nutrition 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 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
- 235000014171 carbonated beverage Nutrition 0.000 description 1
- 150000004649 carbonic acid derivatives Chemical class 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 238000003570 cell viability assay Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 235000013351 cheese Nutrition 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 208000037893 chronic inflammatory disorder Diseases 0.000 description 1
- 235000013985 cinnamic acid Nutrition 0.000 description 1
- 229930016911 cinnamic acid Natural products 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 230000009137 competitive binding Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 229960003638 dopamine Drugs 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000019414 erythritol Nutrition 0.000 description 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 1
- 229940009714 erythritol Drugs 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
- 238000011156 evaluation Methods 0.000 description 1
- 208000030533 eye disease Diseases 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 210000004744 fore-foot Anatomy 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 238000012812 general test Methods 0.000 description 1
- 229960002989 glutamic acid Drugs 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 229960002449 glycine Drugs 0.000 description 1
- 239000000665 guar gum Substances 0.000 description 1
- 235000010417 guar gum Nutrition 0.000 description 1
- 229960002154 guar gum Drugs 0.000 description 1
- 231100000753 hepatic injury Toxicity 0.000 description 1
- 102000054350 human CHI3L1 Human genes 0.000 description 1
- 229940048921 humira Drugs 0.000 description 1
- 235000015243 ice cream Nutrition 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 229960000598 infliximab Drugs 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000024949 interleukin-17 production Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 208000037806 kidney injury Diseases 0.000 description 1
- 229940054136 kineret 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
- 229940069445 licorice extract Drugs 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 229940057995 liquid paraffin Drugs 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 201000003265 lymphadenitis Diseases 0.000 description 1
- 210000004698 lymphocyte Anatomy 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
- 235000013372 meat Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- WBYWAXJHAXSJNI-UHFFFAOYSA-N methyl p-hydroxycinnamate Natural products OC(=O)C=CC1=CC=CC=C1 WBYWAXJHAXSJNI-UHFFFAOYSA-N 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 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
- 235000021243 milk fat Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000002480 mineral oil Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229960005358 monensin Drugs 0.000 description 1
- GAOZTHIDHYLHMS-KEOBGNEYSA-N monensin A Chemical compound C([C@@](O1)(C)[C@H]2CC[C@@](O2)(CC)[C@H]2[C@H](C[C@@H](O2)[C@@H]2[C@H](C[C@@H](C)[C@](O)(CO)O2)C)C)C[C@@]21C[C@H](O)[C@@H](C)[C@@H]([C@@H](C)[C@@H](OC)[C@H](C)C(O)=O)O2 GAOZTHIDHYLHMS-KEOBGNEYSA-N 0.000 description 1
- 235000019462 natural additive Nutrition 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 231100000228 neurotoxicity Toxicity 0.000 description 1
- 230000007135 neurotoxicity Effects 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 239000006186 oral dosage form Substances 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000037361 pathway 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
- 230000002085 persistent effect Effects 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 235000013550 pizza Nutrition 0.000 description 1
- 230000004983 pleiotropic effect Effects 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000010318 polygalacturonic acid Substances 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
- 230000034190 positive regulation of NF-kappaB transcription factor activity 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
- 230000002335 preservative effect Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 1
- 229960003415 propylparaben Drugs 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 208000028172 protozoa infectious disease Diseases 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000010807 real-time PCR kit Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 229940116176 remicade Drugs 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 206010038796 reticulocytosis Diseases 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 235000013580 sausages Nutrition 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 235000011888 snacks Nutrition 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- JUJBNYBVVQSIOU-UHFFFAOYSA-M sodium;4-[2-(4-iodophenyl)-3-(4-nitrophenyl)tetrazol-2-ium-5-yl]benzene-1,3-disulfonate Chemical compound [Na+].C1=CC([N+](=O)[O-])=CC=C1N1[N+](C=2C=CC(I)=CC=2)=NC(C=2C(=CC(=CC=2)S([O-])(=O)=O)S([O-])(=O)=O)=N1 JUJBNYBVVQSIOU-UHFFFAOYSA-M 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 235000014347 soups Nutrition 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 201000004595 synovitis Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 235000013616 tea Nutrition 0.000 description 1
- 150000003526 tetrahydroisoquinolines Chemical class 0.000 description 1
- 229960003989 tocilizumab Drugs 0.000 description 1
- 238000002054 transplantation Methods 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
- 210000004231 tunica media Anatomy 0.000 description 1
- 230000003827 upregulation 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
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/472—Non-condensed isoquinolines, e.g. papaverine
-
- 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
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/306—Foods, ingredients or supplements having a functional effect on health having an effect on bone mass, e.g. osteoporosis prevention
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Rheumatology (AREA)
- General Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Physical Education & Sports Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
Description
본 발명은 테트라하이드로파파베린을 유효성분으로 포함하는 TNF-관련 질환 예방 또는 치료용 조성물 및 이를 이용한 TNF 활성 억제 방법에 관한 것이다.The present invention relates to a composition for preventing or treating TNF-related diseases comprising tetrahydropaberine as an active ingredient, and a method for inhibiting TNF activity using the same.
류마티스 관절염(rheumatoid arthritis; RA)은 주로 관절 조직과 뼈에 영향을 미치는 만성 염증성 질환이다. 류마티스 관절염 환자의 관절에서 활막세포의 활성화는 IL-6, TNF 및 IL-1β를 포함하는 염증성 사이토카인의 생성을 증가시켜 연골과 뼈의 파괴를 유발한다. 상기 사이토카인 중 TNF는 주요한 전염증성 사이토카인으로 활막세포에서 IL-6, IL-8, GM-CSF 및 심지어 그 자체의 생성을 강력하게 유도한다.Rheumatoid arthritis (RA) is a chronic inflammatory disease that primarily affects joint tissues and bones. Activation of synovial cells in the joints of patients with rheumatoid arthritis increases the production of inflammatory cytokines including IL-6, TNF, and IL-1β, causing cartilage and bone destruction. Among these cytokines, TNF is a major pro-inflammatory cytokine and strongly induces the production of IL-6, IL-8, GM-CSF and even itself in synovial cells.
현재, IL-17을 생성하는 Th17이 류마티스 관절염의 발병에 관여한다는 연구 결과가 보고되고 있다. 관절에서 IL-17의 억제 또는 과발현은 각각 관절염을 억제시키거나 악화시킬 수 있다. 건강한 대조군 및 류마티스 관절염 환자로부터 활성화된 단핵구는 시험관 내에서 IL-1β/TNF-α에 의존적으로 Th17 반응을 유도하고 류마티스 관절염의 염증 부위로부터 활성화된 단핵구는 세포 접촉에 의존적으로 증가된 Th17 반응을 유도하는 것으로 보고되었다. 마우스 콜라겐 유도 관절염 모델의 초기 단계에서 IL-17은 TNF-α에 의존하는 반면, 후기 단계에서 IL-17은 주도적이며, TNF-α 독립적으로 작용한다. 비록 류마티스 관절염의 기전이 명확하게 밝혀지지는 않았으나, Th17 반응과 IL-17 생산의 상향 조절이 중요한 역할을 하는 것으로 보고되고 있다.Currently, research results have been reported that Th17 producing IL-17 is involved in the development of rheumatoid arthritis. Inhibition or overexpression of IL-17 in the joint can inhibit or worsen arthritis, respectively. Activated monocytes from healthy controls and rheumatoid arthritis patients induce a Th17 response dependent on IL-1β/TNF-α in vitro, and monocytes activated from the inflammatory site of rheumatoid arthritis induce an increased Th17 response dependent on cell contact. It has been reported to be. In the early stages of the mouse collagen-induced arthritis model, IL-17 is dependent on TNF-α, whereas in the later stages, IL-17 is dominant and acts independently of TNF-α. Although the mechanism of rheumatoid arthritis has not been clearly elucidated, it has been reported that Th17 response and upregulation of IL-17 production play an important role.
TNF-α는 NF-κB 경로의 활성화를 통해 광범위한 전염증 특성을 나타내며, 류마티스 활막염에 결정적으로 관여하는 다면성 사이토카인이다. 예를 들어, TNF-α는 전염증성 사이토카인(IL-1, IL-6 등)과 케모카인(IL-8, MCP-1, MIP-1, RANTES 등)의 합성을 유도하여 대식세포를 활성화시켜 자가면역 병리학에서 전염증성 사이토카인 환경을 지속시킨다. 또한, TNF-α 수용체(TNFR1 및 TNFR2)를 통한 NF-κB의 활성화는 항-세포사멸 단백질을 상향 조절하여 염증 세포의 연장된 생존 및 지속적인 염증을 초래한다.TNF-α exhibits a wide range of proinflammatory properties through activation of the NF-κB pathway, and is a pleiotropic cytokine crucially involved in rheumatoid synovitis. For example, TNF-α activates macrophages by inducing the synthesis of pro-inflammatory cytokines (IL-1, IL-6, etc.) and chemokines (IL-8, MCP-1, MIP-1, RANTES, etc.). Sustaining the pro-inflammatory cytokine environment in autoimmune pathology. In addition, activation of NF-κB through TNF-α receptors (TNFR1 and TNFR2) upregulates anti-apoptotic proteins resulting in prolonged survival and persistent inflammation of inflammatory cells.
1988년에 활성화된 대식세포 또는 림프구에 의해 분비된 사이토카인인 TNF를 표적으로 하는 생물학적 제제가 개발되었다. 항-TNF 바이오 신약에는 etanercept(Enbrel), infliximab(Remicade), adalimumab(Humira) 등이 있고, 상이한 기전을 갖는 항염증성 바이오 신약 중 tocilizumab(Actemra)은 신호 전달을 차단하기 위해 IL-6 수용체에 결합하는 모노클로날 항체이며, Anakinra(Kineret)는 IL-1 수용체에 결합하는 길항제이다. 상기 바이오 신약은 단기간에 효과를 나타내지만 반복 주사 치료로 인한 잦은 병원 방문, 높은 저항력, 내성 면역 부작용, 고비용, 저온 저장 등의 단점을 가진다. 따라서, TNF에 직접 결합하여 류마티스 관절염과 같은 자가면역질환을 치료할 수 있는 소분자 화합물의 개발이 필요한 실정이다.In 1988, a biological agent targeting TNF, a cytokine secreted by activated macrophages or lymphocytes, was developed. New anti-TNF biologic drugs include etanercept (Enbrel), infliximab (Remicade), and adalimumab (Humira), and among anti-inflammatory biologic drugs with different mechanisms, tocilizumab (Actemra) binds to the IL-6 receptor to block signal transduction. Is a monoclonal antibody, and Anakinra (Kineret) is an antagonist that binds to the IL-1 receptor. The new bio-drug is effective in a short period of time, but has disadvantages such as frequent hospital visits due to repeated injection treatment, high resistance, immune side effects, high cost, and low temperature storage. Therefore, there is a need to develop a small molecule compound capable of directly binding to TNF to treat autoimmune diseases such as rheumatoid arthritis.
테트라하이드로파파베린(tetrahydropapaverine; THP)은 테트라하이드로이소 퀴놀린 중 하나로, 살소리롤(salsolinol) 및 테트라하이드로파파베롤린의 유사체이며, 도파민 뉴런에 신경 독성을 나타내는 것으로 알려져 있다. TNF 억제 효과 또는 자가면역질환 치료에 있어서 테트라하이드로파파베린의 효과는 아직까지 밝혀진 바가 없다.Tetrahydropapaverine (THP) is one of the tetrahydroisoquinolines, an analogue of salsorol and tetrahydropaberolin, and is known to exhibit neurotoxicity to dopamine neurons. The TNF inhibitory effect or the effect of tetrahydropaberine in the treatment of autoimmune diseases has not yet been revealed.
본 발명의 목적은 TNF-관련 질환 예방 또는 치료용 약학 조성물을 제공하는 데에 있다.An object of the present invention is to provide a pharmaceutical composition for preventing or treating TNF-related diseases.
본 발명의 다른 목적은 TNF-관련 질환 예방 또는 개선용 건강식품 조성물을 제공하는 데에 있다.Another object of the present invention is to provide a health food composition for preventing or improving TNF-related diseases.
본 발명의 또 다른 목적은 TNF-관련 질환을 치료하는 방법을 제공하는 데에 있다.Another object of the present invention is to provide a method for treating TNF-related diseases.
본 발명의 또 다른 목적은 인 비트로(in vitro)에서 TNF 활성을 억제하는 시약 조성물을 제공하는 데에 있다.Another object of the present invention is to provide a reagent composition that inhibits TNF activity in vitro.
본 발명의 또 다른 목적은 TNF 활성 억제 방법을 제공하는 데에 있다.Another object of the present invention is to provide a method for inhibiting TNF activity.
상기 목적을 달성하기 위하여, 본 발명은 테트라하이드로파파베린(tetrahydropapaverine)을 유효성분으로 포함하는 TNF-관련 질환 예방 또는 치료용 약학 조성물을 제공한다.In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating TNF-related diseases comprising tetrahydropapaverine as an active ingredient.
또한, 본 발명은 테트라하이드로파파베린(tetrahydropapaverine)을 유효성분으로 포함하는 TNF-관련 질환 예방 또는 개선용 건강식품 조성물을 제공한다.In addition, the present invention provides a health food composition for preventing or improving TNF-related diseases comprising tetrahydropapaverine as an active ingredient.
또한, 본 발명은 테트라하이드로파파베린(tetrahydropapaverine)을 약학적으로 유효한 양으로 처리하는 단계를 포함하는, TNF-관련 질환을 치료하는 방법을 제공한다.In addition, the present invention provides a method for treating TNF-related diseases, comprising the step of treating tetrahydropapaverine in a pharmaceutically effective amount.
또한, 본 발명은 테트라하이드로파파베린(tetrahydropapaverine)을 유효성분으로 포함하며, 인 비트로(in vitro)에서 TNF 활성을 억제하는 시약 조성물을 제공한다.In addition, the present invention provides a reagent composition comprising tetrahydropapaverine as an active ingredient and inhibiting TNF activity in vitro.
또한, 본 발명은 사람을 제외한 동물에 테트라하이드로파파베린(tetrahydropapaverine)을 처리하는 단계를 포함하는, TNF 활성 억제 방법을 제공한다.In addition, the present invention provides a method for inhibiting TNF activity, comprising the step of treating tetrahydropapaverine to animals other than humans.
본 발명에서는 천연 화합물 라이브러리를 스크리닝하여 TNF의 직접 결합제 및 억제제로 작용하는 테트라하이드로파파베린을 동정하였다. 상기 테트라하이드로파파베린은 TNF 활성을 억제하고, 콜라겐 유도 관절염 모델에서 뼈 및 연골을 보호하며, 염증성 사이토카인을 감소시킬 뿐만 아니라 Th17 세포의 분화 및 섬유아세포 유사 활막세포의 증식을 억제하는 효과를 나타내는 바, 상기 테트라하이드로파파베린은 류마티스 관절염을 포함하는 TNF-관련 질환의 예방 또는 치료용 약학 조성물, TNF-관련 질환의 예방 또는 개선용 건강식품 조성물 등으로 유용하게 활용될 수 있다. In the present invention, a natural compound library was screened to identify tetrahydropaberine acting as a direct binding agent and inhibitor of TNF. The tetrahydropaberine inhibits TNF activity, protects bone and cartilage in a collagen-induced arthritis model, reduces inflammatory cytokines, and inhibits the differentiation of Th17 cells and proliferation of fibroblast-like synovial cells. Bar, the tetrahydropaberine may be usefully used as a pharmaceutical composition for preventing or treating TNF-related diseases including rheumatoid arthritis, a health food composition for preventing or improving TNF-related diseases, and the like.
도 1은 TNF-TNF 수용체의 결합을 억제하는 소분자 화합물 검출에 관한 것으로, (A)는 TNF-TNF 수용체 결합 ELISA를 이용한 천연 단일 화합물의 스크리닝, (B)는 테트라하이드로파파베린의 화학 구조, (C)는 TNF에 대한 THP의 직접 결합 활성, (D)는 LM 세포에서 THP의 TNF 특이적 중화 활성을 나타낸 결과이다.
도 2는 콜라겐 유도 관절염(CIA) 모델에서 THP의 치료 효과에 관한 것으로, (A)는 관절염 점수, (B)는 혈청 사이토카인 수준, (C)는 조직학적 이미지를 나타낸 결과이다.
도 3은 마우스 비장세포를 이용한 Th17 세포의 분화 및 기능에서 THP의 억제 효과에 관한 것으로, (A)는 유세포 분석을 이용한 IL-17A의 발현, (B)는 ELISA 및 qRT-PCR을 이용한 IL-17A의 발현, (C)는 qRT-PCR을 이용한 RORγt의 발현을 나타낸 결과이다.
도 4는 인간 PBMC를 이용한 Th17 세포의 분화 및 기능에서 THP의 억제 효과에 관한 것으로, (A)는 유세포 분석을 이용한 IL-17A의 발현, (B)는 ELISA 및 qRT-PCR을 이용한 IL-17A의 발현, (C)는 qRT-PCR을 이용한 RORγt의 발현을 나타낸 결과이다.
도 5는 MH7A 세포에서 THP에 의한 세포 증식 억제 활성을 나타낸 결과이다.Figure 1 relates to the detection of a small molecule compound that inhibits the binding of TNF-TNF receptor, (A) is a screening of a natural single compound using a TNF-TNF receptor binding ELISA, (B) is the chemical structure of tetrahydropaberine, ( C) is the direct binding activity of THP to TNF, (D) is the result showing the TNF-specific neutralizing activity of THP in LM cells.
FIG. 2 shows the therapeutic effect of THP in a collagen-induced arthritis (CIA) model, where (A) is an arthritis score, (B) is a serum cytokine level, and (C) is a histological image.
Figure 3 relates to the inhibitory effect of THP on the differentiation and function of Th17 cells using mouse splenocytes, (A) is the expression of IL-17A using flow cytometry, (B) is IL- using ELISA and qRT-PCR. Expression of 17A, (C) is a result showing the expression of RORγt using qRT-PCR.
Figure 4 relates to the inhibitory effect of THP on the differentiation and function of Th17 cells using human PBMC, (A) is the expression of IL-17A using flow cytometry, (B) is IL-17A using ELISA and qRT-PCR. Expression of, (C) is a result showing the expression of RORγt using qRT-PCR.
5 is a result showing the cell proliferation inhibitory activity by THP in MH7A cells.
본 발명의 발명자들은 천연 단일 화합물 라이브러리에서 TNF 결합 스크리닝을 통해 테트라하이드로파파베린을 동정하였으며, 상기 테트라하이드로파파베린이 TNF에 결합하여 TNF 활성을 억제하고, 염증성 사이토카인의 분비, Th17 세포의 분화, Th17 세포와 관련된 분자의 발현, FLS 세포의 증식을 억제하여 류마티스 관절염을 개선시키는 것을 확인하며 본 발명을 완성하였다.The inventors of the present invention identified tetrahydropaberine through TNF binding screening in a natural single compound library, and the tetrahydropaberin binds to TNF and inhibits TNF activity, secretion of inflammatory cytokines, differentiation of Th17 cells, The present invention was completed by confirming the improvement of rheumatoid arthritis by inhibiting the expression of molecules related to Th17 cells and proliferation of FLS cells.
이에, 본 발명은 테트라하이드로파파베린(tetrahydropapaverine)을 유효성분으로 포함하는 TNF-관련 질환 예방 또는 치료용 약학 조성물을 제공한다.Accordingly, the present invention provides a pharmaceutical composition for preventing or treating TNF-related diseases comprising tetrahydropapaverine as an active ingredient.
상기 테트라하이드로파파베린은 종양괴사인자(TNF)에 직접 결합하여 TNF 활성을 억제할 수 있다.The tetrahydropaberine can inhibit TNF activity by directly binding to tumor necrosis factor (TNF).
상기 TNF-관련 질환은 자가 면역 질환, 염증성 질환, 심혈관 질환, 대사성 질환, 면역 장애, 신경 질환, 안과 질환, 피부 질환, 정신 질환, 감염 질환 및 암으로 이루어진 군에서 선택되는 어느 하나일 수 있으나, 이에 제한되는 것은 아님을 명시한다.The TNF-related disease may be any one selected from the group consisting of autoimmune diseases, inflammatory diseases, cardiovascular diseases, metabolic diseases, immune disorders, neurological diseases, eye diseases, skin diseases, mental diseases, infectious diseases and cancer, It should be noted that it is not limited thereto.
상세하게는, 상기 TNF-관련 질환은 류마티스 관절염, 소아류마티스관절염, 염증성 장질환, 크론병, 궤양성 대장염, 건선, 판상건선, 소아판상건선, 건선성관절염, 다관절형소아특발성관절염, 베체트장염, 강직성척추염, 축성척추관절염, 소아골부착부위염관련관절염, 류마티스다발근통, 다발성경화증, 갑상선염, 지연과민증, 알레르기, 접촉성피부염, 아토피성피부염, 전신성홍반루푸스, 전신경화증, 성인형스틸병, 천식, 자가면역성갑상선장애, 쇼그렌증후군, 가와사키병, 췌장염, 신장염, 간염, 폐렴, 만성폐쇄성폐질환, 중이염, 맥관증식신염, 골수형성이상증후군, 골관절염, 유육종증, 고리육아종, 베게너육아종증, 낭창, 용혈성요독증후군, 동맥경화증, 혈관염, 심부전, 뇌졸중, 심근경색, 심근허혈-재관류손상, 성기능장애, 비만, 고혈압, 당뇨병 및 당뇨합병증, 고지혈증, 자간전증, 신장병, 간장병, 신장 손상, 간손상, 뱀교상, 동종이식거부, 장기이식, 이식편대숙주병, 치매, 알츠하이머병, 파킨슨병, 근위축성측색경화증, 통증, 중추신경계질환, 포도막염, 베체트병, 당뇨황반부종, 황반변성, 안와병증, 녹내장, 화농성한선염, 다중심성망상조직구증식증, 모공성홍색비강진, 호산구성근막염, 지방층염, 당뇨병성유지방성괴사생성, 흉터유사천포창, 괴저성농피증, 스위트증후군, 각질하농포성피부증, 피부경화증, 호중구성피부염, 독성표피괴사융해, 농포성피부염, 피부근염, 다발근육염, 물집피부병, 결절홍반, 탈모증, 우울증, 양극성장애, 불안장애, 결핵, 바이러스감염, 박테리아감염, 진균감염, 원충감염, 뇌말라리아, 패혈증, 패혈성 쇼크, 전립선암, 피부암, 대장암, 신장암, 췌장암, 난소암, 유방암, 방광암, 전립선암, 림프종, 교종, 골육종, 백혈병, 다발성골수종 및 악액질로 이루어진 군에서 선택될 수 있으나, 이에 제한되는 것은 아님을 명시한다.Specifically, the TNF-related diseases are rheumatoid arthritis, juvenile rheumatoid arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, psoriasis, plaque psoriasis, juvenile plaque psoriasis, psoriatic arthritis, polyarticular pediatric idiopathic arthritis, Behcet's enteritis. , Ankylosing spondylitis, axial spondyloarthritis, pediatric bone-attached gastritis-related arthritis, rheumatoid polymyalgia, multiple sclerosis, thyroiditis, delayed hypersensitivity, allergies, contact dermatitis, atopic dermatitis, systemic lupus erythematosus, systemic sclerosis, adult steel disease, asthma , Autoimmune thyroid disorder, Sjogren's syndrome, Kawasaki's disease, pancreatitis, nephritis, hepatitis, pneumonia, chronic obstructive pulmonary disease, otitis media, vascular proliferative nephritis, myelodysplastic syndrome, osteoarthritis, sarcoidosis, ring granuloma, Wegener's granulomatosis, lupus, hemolytic Uremic syndrome, arteriosclerosis, vasculitis, heart failure, stroke, myocardial infarction, myocardial ischemia-reperfusion injury, sexual dysfunction, obesity, hypertension, diabetes and diabetes complications, hyperlipidemia, preeclampsia, kidney disease, liver disease, kidney injury, liver injury, snake bite, Allograft rejection, organ transplantation, graft versus host disease, dementia, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, pain, central nervous system disease, uveitis, Behcet's disease, diabetic macular edema, macular degeneration, orbitopathy, glaucoma, purulent Adenitis, multicentric reticulocytosis, follicular erythema, eosinophilic fasciitis, steatitis, diabetic milk fat necrosis, scar-like pemphigus, gangrene pyoderma, sweet syndrome, subkeratinous pustular dermatosis, scleroderma, neutrophilic dermatitis , Toxic epidermal necrosis fusion, pustular dermatitis, dermatitis, polymyositis, blister dermatitis, nodular erythema, alopecia, depression, bipolar disorder, anxiety disorder, tuberculosis, viral infection, bacterial infection, fungal infection, protozoal infection, cerebral malaria, sepsis , Septic shock, prostate cancer, skin cancer, colon cancer, kidney cancer, pancreatic cancer, ovarian cancer, breast cancer, bladder cancer, prostate cancer, lymphoma, glioma, osteosarcoma, leukemia, multiple myeloma, and cachexia, but It should be noted that it is not limited.
상기 조성물은 TNF, IL-6 및 IL-1β의 발현을 감소시킬 수 있고, Th17 세포의 분화 및 섬유아세포 유사 활막세포의 증식을 억제시킬 수 있다.The composition can reduce the expression of TNF, IL-6 and IL-1β, and can inhibit the differentiation of Th17 cells and the proliferation of fibroblast-like synovial cells.
본 발명의 조성물이 약학 조성물인 경우, 투여를 위하여, 상기 기재한 유효성분 이외에 약학적으로 허용 가능한 담체, 부형제 또는 희석제를 포함할 수 있다. 상기 담체, 부형제 및 희석제로는 락토오스, 덱스트로오스, 수크로오스, 소르비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로오스, 메틸 셀룰로오스, 미정질 셀룰로오스, 폴리비닐피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다.When the composition of the present invention is a pharmaceutical composition, for administration, it may include a pharmaceutically acceptable carrier, excipient, or diluent in addition to the above-described active ingredients. Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oils.
본 발명의 약학 조성물은 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 또는 멸균 주사용액의 형태로 제형화하여 사용할 수 있다. 상세하게는 제형화할 경우 통상 사용하는 충진제, 중량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제될 수 있다. 경구투여를 위한 고형 제제로는 정제, 환제, 산제, 과립제, 캡슐제 등을 포함하나, 이에 한정되는 것은 아니다. 이러한 고형 제제는 상기 유효성분 외에 적어도 하나 이상의 부형제, 예를 들면, 전분, 칼슘 카보네이트, 수크로오스, 락토오스, 젤라틴 등을 섞어 조제될 수 있다. 또한, 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용될 수 있다. 경구를 위한 액상물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등을 첨가하여 조제될 수 있다. 비경구 투여를 위한 제제는 멸균된 수용액, 비수성 용제, 현탁제, 유제, 동결건조 제제 및 과제를 포함한다. 비수성 용제 및 현탁제로는 프로필렌글리콜, 폴리에틸렌글리콜, 올리브 오일과 같은 식물성 오일, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔, 마크로솔, 트윈 61, 카카오지, 라우린지, 글리세로젤라틴 등이 사용될 수 있다.The pharmaceutical compositions of the present invention can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, or sterile injectable solutions according to a conventional method. . Specifically, when formulated, it may be prepared using diluents or excipients such as fillers, weight agents, binders, wetting agents, disintegrants, and surfactants that are commonly used. Solid preparations for oral administration include, but are not limited to, tablets, pills, powders, granules, capsules, and the like. Such a solid preparation may be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose, lactose, gelatin, etc. in addition to the active ingredient. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. It can be prepared by adding various excipients, such as wetting agents, sweetening agents, fragrances, preservatives, and the like, in addition to oral liquids and liquid paraffin. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations and tasks. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like may be used. As a base for suppositories, witepsol, macrosol, Tween 61, cacao butter, laurin paper, glycerogelatin, and the like can be used.
본 발명의 약학 조성물의 적합한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 시간에 따라 다르지만, 당 업자에 의해 적절하게 선택될 수 있는 바, 상기 조성물의 일일 투여량은 바람직하게는 0.001 mg/kg 내지 50 mg/kg이며, 필요에 따라 일일 1회 내지 수회로 나누어 투여할 수 있다.A suitable dosage of the pharmaceutical composition of the present invention varies depending on the condition and weight of the patient, the severity of the disease, the form of the drug, and the time, but may be appropriately selected by a person skilled in the art. The daily dosage of the composition is preferably It is 0.001 mg/kg to 50 mg/kg, and it can be administered once to several times a day, if necessary.
또한, 본 발명은 테트라하이드로파파베린(tetrahydropapaverine)을 유효성분으로 포함하는 TNF-관련 질환 예방 또는 개선용 건강식품 조성물을 제공한다.In addition, the present invention provides a health food composition for preventing or improving TNF-related diseases comprising tetrahydropapaverine as an active ingredient.
본 발명의 조성물이 건강식품 조성물인 경우, 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 천연 과일 주스, 합성 과일 주스 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 또한, 건강식품 조성물은 육류, 소세지, 빵, 초콜릿, 캔디류, 스넥류, 과자류, 피자, 라면, 껌류, 아이스크림류, 스프, 음료수, 차, 기능수, 드링크제, 알코올 및 비타민 복합제 중 어느 하나의 형태일 수 있다.When the composition of the present invention is a health food composition, various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and salts thereof , Alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, and the like. In addition, it may contain pulp for the manufacture of natural fruit juices, synthetic fruit juices and vegetable beverages. These components may be used independently or in combination. In addition, the health food composition may be in the form of any one of meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, gum, ice cream, soup, beverage, tea, functional water, drink, alcohol and vitamin complex. I can.
또한, 상기 건강식품 조성물은 식품첨가물을 추가로 포함할 수 있으며, 식품첨가물로서의 적합 여부는 다른 규정이 없는 한 식품의약품안전처에 승인된 식품첨가물공전의 총칙 및 일반 시험법 등에 따라 해당 품목에 관한 규격 및 기준에 의하여 판정한다.In addition, the health food composition may additionally contain food additives, and the suitability as a food additive is determined according to the general rules and general test methods of the Food Additive Code approved by the Ministry of Food and Drug Safety, unless otherwise specified. It is judged according to the standards and standards.
상기 식품첨가물공전에 수재된 품목으로 예를 들어, 케톤류, 글리신, 구연산 칼륨, 니코틴산, 계피산 등의 화학적 합성품, 감색소, 감초추출물, 결정셀룰로오스, 고랭색소, 구아검 등의 천연첨가물, L-글루타민산나트륨 제제, 면류 첨가 알칼리제, 보존료제제, 타르색소 제제 등의 혼합 제제류 등을 들 수 있다.For example, ketones, glycine, potassium citrate, nicotinic acid, cinnamic acid, chemical synthetic products such as reduced pigment, licorice extract, crystalline cellulose, high cooling pigment, natural additives such as guar gum, L-glutamic acid And mixed preparations such as sodium preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations.
이때, 건강식품 조성물을 제조하는 과정에서 식품에 첨가되는 본 발명에 따른 조성물은 필요에 따라 그 함량을 적절히 가감할 수 있다.At this time, the content of the composition according to the present invention added to food in the process of manufacturing the health food composition may be appropriately added or subtracted as needed.
또한, 본 발명은 테트라하이드로파파베린(tetrahydropapaverine)을 약학적으로 유효한 양으로 처리하는 단계를 포함하는, TNF-관련 질환을 치료하는 방법을 제공한다.In addition, the present invention provides a method for treating TNF-related diseases, comprising the step of treating tetrahydropapaverine in a pharmaceutically effective amount.
또한, 본 발명은 테트라하이드로파파베린(tetrahydropapaverine)을 유효성분으로 포함하며, 인 비트로(in vitro)에서 TNF 활성을 억제하는 시약 조성물을 제공한다.In addition, the present invention provides a reagent composition comprising tetrahydropapaverine as an active ingredient and inhibiting TNF activity in vitro.
또한, 본 발명은 사람을 제외한 동물에 테트라하이드로파파베린(tetrahydropapaverine)을 처리하는 단계를 포함하는, TNF 활성 억제 방법을 제공한다.In addition, the present invention It provides a method for inhibiting TNF activity, comprising the step of treating an animal other than humans with tetrahydropapaverine.
이하에서는 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail through examples. These examples are only for describing the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .
실시예 1: 마우스Example 1: Mouse
6주령 수컷 DBA/1J 또는 C57BL/6 마우스는 Orientbio(Gyeonggi-do, Korea)에서 구입하였다. 마우스는 12시간 명/암 주기, 온도(22 ± 3℃) 및 습도(40-60%)가 조절된 환경에서 사육하였으며, 식이 및 물은 자유롭게 섭취할 수 있게 하였다. 마우스는 1주일의 순응 기간을 거친 후 실험에 사용하였다. 모든 동물실험은 가톨릭대학교 동물실험윤리위원회(IACUC)가 승인한 프로토콜(허가번호: 2016-018-02)을 준수하였다.6-week-old male DBA/1J or C57BL/6 mice were purchased from Orientbio (Gyeonggi-do, Korea). Mice were bred in an environment in which 12 hours light/dark cycle, temperature (22 ± 3°C) and humidity (40-60%) were controlled, and diet and water were freely ingested. Mice were used in the experiment after an acclimatization period of 1 week. All animal experiments followed the protocol (license number: 2016-018-02) approved by the Catholic University's Animal Experimental Ethics Committee (IACUC).
실시예 2: 콜라겐 유도 관절염 동물모델 제작 및 테트라하이드로파파베린(THP) 주사Example 2: Collagen-induced arthritis animal model production and tetrahydropaberine (THP) injection
콜라겐 유도 관절염(collagen-induced arthritis; CIA)은 하기와 같이 유도되었다. 간략하게, 소 타입 II 콜라겐(CII)(Chondrex, ES) 및 Complete Freund’s adjuvant(Sigma, US) 50 μg를 꼬리 기저에 피내 주사하였다. 2주 후, CII 및 Incomplete Freund’s adjuvant(Sigma, US) 50 μg를 피내 주사하여 부스팅하였다. THP(5 mg/kg), 또는 비히클 대조군(10% DMSO가 포함된 증류수)은 첫 면역 후 CIA 마우스(N = 10)에 주 6회 경구 투여하였다.Collagen-induced arthritis (CIA) was induced as follows. Briefly, 50 μg of bovine type II collagen (CII) (Chondrex, ES) and Complete Freund's adjuvant (Sigma, US) were injected intradermally at the base of the tail. After 2 weeks, 50 μg of CII and Incomplete Freund's adjuvant (Sigma, US) was boosted by intradermal injection. THP (5 mg/kg) or vehicle control (distilled water containing 10% DMSO) was orally administered 6 times a week to CIA mice (N = 10) after the first immunization.
실시예 3: CIA의 임상 관절염 점수 평가Example 3: Evaluation of CIA's Clinical Arthritis Score
마우스의 4개 발에서 관절염의 중증도는 하기의 방법을 이용하여 3명의 맹검 관찰자에 의해 평가되었다. 앞발 및 뒷발의 관절염의 중증도는 첫 면역 후 0 내지 56일 사이에 주 2회 평가하였다: 0 = 정상 발, 1 = 1개의 발에 염증이 있고 부어오름, 2 = 1~3개의 발에 염증이 있고 부어오름, 3 = 발 전체에 염증이 있고 부어오름, 4 = 염증이 심하고 부어오르거나 유착된 발. 각 마우스의 관절염 점수는 4개 발의 관절염 중증도의 합으로 나타내었다. 가장 높은 점수는 16점이다.The severity of arthritis in the four paws of mice was evaluated by three blind observers using the following method. The severity of arthritis of the forefoot and hind paw was assessed twice a week between 0 and 56 days after the first immunization: 0 = normal foot, 1 = 1 foot is inflamed and swollen, 2 = 1 to 3 feet are inflamed. And swollen, 3 = inflamed and swollen all over the foot, 4 = inflamed, swollen or adherent foot. The arthritis score of each mouse was expressed as the sum of the arthritis severity of the four paws. The highest score is 16 points.
실시예 4: 조직학적 분석Example 4: Histological analysis
뒷발을 절개하고 10% 포름산으로 21일 동안 탈석회 과정을 수행하였다. 탈석회 후, 표본을 파라핀에 포매시킨 후 조직 절편을 제작하였다. 조직 절편은 탈파라핀 후 재수화시킨 다음, 헤마톡실린 & 에오신, 또는 사프라닌 O로 염색하였다.The hind paw was dissected and the deliming process was performed for 21 days with 10% formic acid. After deliming, the specimen was embedded in paraffin, and a tissue section was prepared. Tissue sections were rehydrated after deparaffining, and then stained with hematoxylin & eosin, or safranin O.
실시예 5: 생체 외 마우스 Th17 세포 분화Example 5: In vitro mouse Th17 cell differentiation
naive 마우스를 희생시킨 후, 비장을 적출하여 단일세포로 분리하였다. 이후 세포를 얼음 위에 놓고 3분 동안 RBC 용해 완충액(Thermo, US)과 반응시켰다. PBS로 세척한 후, 마우스 CD4 MACS 마이크로비드(MiltenyiBiotec, Germany)를 사용하여 naive CD4+ T 세포를 분리하였다.After the naive mouse was sacrificed, the spleen was excised and separated into single cells. Thereafter, the cells were placed on ice and reacted with RBC lysis buffer (Thermo, US) for 3 minutes. After washing with PBS, naive CD4 + T cells were isolated using mouse CD4 MACS microbeads (MiltenyiBiotec, Germany).
마우스 Th17 분화 키트(R&D system, US)를 사용하여 마우스 비장세포에 대한 생체 외 실험을 수행하였다. naive CD4+ T 세포(1 X 106 세포/ml)를 마우스 Th17 분화 배지에 현탁시켰다. 부유 세포(1 X 105 세포/웰)를 96 웰 플레이트(Gibco, US)에 접종하고 미리 코팅된 햄스터 항-CD3 마우스 항체로 자극시켰다. 이들 세포를 37℃, 5% CO2 배양기에서 3일 동안 배양하였다. 3일째에 동일한 부피의 새로운 마우스 Th17 분화 배지를 첨가하여 Th17 분화 배지를 교체하고, 2일 동안 추가 배양하였다. 이후, 세포 및 배양 상층액을 회수하고, 수집된 세포는 유세포 분석을 이용하여 Th17 세포 분화 여부를 측정하였다. 또한, 배양 상층액은 IL-17A ELISA 키트(Biolegend, US)를 사용하여 마우스 IL-17A 분비를 측정하였다.In vitro experiments were performed on mouse splenocytes using a mouse Th17 differentiation kit (R&D system, US). naive CD4 + T cells (1
실시예 6: 인간 말초혈액 단핵세포(PBMC)에서 CD4Example 6: CD4 in human peripheral blood mononuclear cells (PBMC) ++ T 세포 분리 및 Th17 세포 분화 T cell isolation and Th17 cell differentiation
건강한 기증자 3명의 시료는 적십자로부터 제공받았으며, TrimaAccel 자동 혈액 수집 시스템을 사용하여 혈액을 수집하였다. 키트에 부착된 LRS(Leukoreduction system) 챔버는 상용화 제품을 구입하여 사용하였다. LRS 챔버에 연결된 input 및 output 튜브는 알코올로 멸균하고 절단하여 사용하였다. 세포를 50 ml 튜브에 넣고, 동량의 2% 우태아혈청(FBS, Corning, US)이 포함된 인산완충식염수(PBS, Welgene)를 첨가하였다. 이어서 SepMateTM-50 튜브(STEMCELL technology, Canada)에 Ficoll-PoqueTM PLUS ratio 1.077(GE Healthcare, US) 15 ml을 첨가하고, 그 위에 LRS 챔버로 분리된 15 ml의 세포를 포개어 넣었다. 브레이크 오프 상태에서 1200 g로 20분 동안 원심분리한 후, 말초혈액 단핵세포를 새로운 50 ml 튜브로 옮겼으며, 2% FBS가 포함된 PBS로 2회 세척하였다.Samples from three healthy donors were provided by the Red Cross, and blood was collected using the TrimaAccel automated blood collection system. The LRS (Leukoreduction system) chamber attached to the kit was used by purchasing a commercial product. The input and output tubes connected to the LRS chamber were sterilized with alcohol and cut. The cells were placed in a 50 ml tube, and phosphate buffered saline (PBS, Welgene) containing an equal amount of 2% fetal calf serum (FBS, Corning, US) was added. Subsequently, 15 ml of Ficoll-Poque™ PLUS ratio 1.077 (GE Healthcare, US) was added to a SepMate™-50 tube (STEMCELL technology, Canada), and 15 ml of cells separated by an LRS chamber were superimposed thereon. After centrifugation at 1200 g for 20 minutes in the break off state, the peripheral blood mononuclear cells were transferred to a new 50 ml tube, and washed twice with PBS containing 2% FBS.
인간 PBMC를 얼음 위에 놓고 5분 동안 RBC 용해 완충액(Thermo, US)과 반응시켰다. PBS로 세척한 후, 인간 CD4 MACS 마이크로비드(MiltenyiBiotec, Germany)를 사용하여 naive CD4+ T 세포를 분리하였다. 인간 Th17 분화 키트(R&D system, US)를 사용하여 PBMC에 대한 생체 외 실험을 수행하였다. 수집된 세포는 유세포 분석을 이용하여 Th17 세포 분화 여부를 측정하였다. 또한, 배양 상층액은 IL-17A ELISA 키트(Biolegend, US)를 사용하여 인간 IL-17A 분비를 측정하였다.Human PBMCs were placed on ice and reacted with RBC lysis buffer (Thermo, US) for 5 minutes. After washing with PBS, naive CD4 + T cells were isolated using human CD4 MACS microbeads (MiltenyiBiotec, Germany). In vitro experiments were performed on PBMCs using the human Th17 differentiation kit (R&D system, US). The collected cells were measured for Th17 cell differentiation using flow cytometry. In addition, the culture supernatant was measured for human IL-17A secretion using an IL-17A ELISA kit (Biolegend, US).
실시예 7: 생체 외 IL-17A 분비 분석Example 7: In vitro IL-17A secretion assay
ELISA 플레이트를 마우스 또는 인간 IL-17A 포획 항체가 포함된 PBS로 4℃에서 밤새 코팅하였다. 코팅 후, 플레이트를 PBST(0.05% 트윈-20이 포함된 PBS)로 세척하고, 1% BSA가 포함된 PBS와 상온에서 진탕하면서 1시간 동안 반응시켜 블로킹하였다. PBST로 세척한 후, PBSA(1% BSA가 포함된 PBS)로 희석된 배양 상층액을 각 웰에 첨가하였다. 플레이트를 4℃에서 진탕하면서 밤새 반응시켰다. 세척 후, 1X 검출 항체를 각 웰에 첨가하고, 플레이트를 상온에서 진탕하면서 1시간 동안 반응시켰다. PBST로 세척한 후, 검출 항체(avidin-HRP)를 첨가하고 플레이트를 상온에서 진탕하면서 1시간 동안 반응시켰다. 세척 후, TMB 용액을 각 웰에 첨가하고 플레이트를 실온에서 진탕하면서 반응시켰다. 이후, 정지 용액(2N HCl)을 각 웰에 첨가하고, 마이크로플레이트 리더기를 이용하여 450 nm에서 흡광도를 측정하였다.ELISA plates were coated overnight at 4° C. with PBS containing mouse or human IL-17A capture antibody. After coating, the plate was washed with PBST (PBS containing 0.05% Tween-20), and blocked by reacting with PBS containing 1% BSA for 1 hour while shaking at room temperature. After washing with PBST, a culture supernatant diluted with PBSA (PBS containing 1% BSA) was added to each well. The plate was reacted overnight while shaking at 4°C. After washing, 1X detection antibody was added to each well, and the plate was reacted for 1 hour while shaking at room temperature. After washing with PBST, a detection antibody (avidin-HRP) was added, and the plate was reacted for 1 hour while shaking at room temperature. After washing, TMB solution was added to each well and the plate was reacted with shaking at room temperature. Thereafter, a stop solution (2N HCl) was added to each well, and absorbance was measured at 450 nm using a microplate reader.
실시예 8: 효소면역분석법Example 8: Enzyme Immunoassay
CIA 마우스에서 TNF, IL-6 및 IL-1β 분비를 확인하기 위해, CIA 실험 56일째에 마우스로부터 혈청을 채취하였다. ELISA 플레이트(Nunc, Denmark)를 1X 마우스 TNF, IL-6 또는 IL-1β 포획 항체가 포함된 PBS로 4℃에서 밤새 코팅하였다. 코팅 후, 플레이트를 PBST(0.05% 트윈-20이 포함된 PBS)로 세척하고, 1% BSA가 포함된 PBS와 상온에서 진탕하면서 1시간 동안 반응시켜 블로킹하였다. PBST로 세척한 후, PBSA(1% BSA가 포함된 PBS)로 희석된 배양 상층액을 각 웰에 첨가하였다. 플레이트를 4℃에서 진탕하면서 밤새 반응시켰다. 세척 후, 1X 검출 항체를 각 웰에 첨가하고, 플레이트를 상온에서 진탕하면서 1시간 동안 반응시켰다. PBST로 세척한 후, 검출 항체(avidin-HRP)를 첨가하고 플레이트를 상온에서 진탕하면서 1시간 동안 반응시켰다. 세척 후, TMB 용액을 각 웰에 첨가하고 플레이트를 실온에서 진탕하면서 반응시켰다. 이후, 정지 용액(2N HCl)을 각 웰에 첨가하고, 마이크로플레이트 리더기를 이용하여 450 nm에서 흡광도를 측정하였다.In order to confirm the secretion of TNF, IL-6 and IL-1β in CIA mice, serum was collected from mice on day 56 of the CIA experiment. ELISA plates (Nunc, Denmark) were coated overnight at 4° C. with PBS containing 1X mouse TNF, IL-6 or IL-1β capture antibody. After coating, the plate was washed with PBST (PBS containing 0.05% Tween-20), and blocked by reacting with PBS containing 1% BSA for 1 hour while shaking at room temperature. After washing with PBST, a culture supernatant diluted with PBSA (PBS containing 1% BSA) was added to each well. The plate was reacted overnight while shaking at 4°C. After washing, 1X detection antibody was added to each well, and the plate was reacted for 1 hour while shaking at room temperature. After washing with PBST, a detection antibody (avidin-HRP) was added, and the plate was reacted for 1 hour while shaking at room temperature. After washing, TMB solution was added to each well and the plate was reacted with shaking at room temperature. Thereafter, a stop solution (2N HCl) was added to each well, and absorbance was measured at 450 nm using a microplate reader.
실시예 9: 유세포 분석Example 9: Flow cytometry
수집된 마우스 또는 인간 Th17 세포를 ep 튜브에 넣고 원심분리한 후 PBS로 세척하였다. 세척 후, 단일세포를 1X stimulation cocktail 및 1X monensin과 37℃에서 4시간 동안 자극시켰다. 자극된 세포를 원심분리하고, 항-마우스 또는 인간 CD4-PEcy7 염색 완충액과 빛 차단 하에 4℃에서 30분 동안 반응시켰다. 세척 후, 시료에 fixation/permeabilization 용액을 첨가하고 빛 차단 하에 4℃에서 18시간 동안 반응시켰다. 세척 후, 세포를 항-마우스 또는 인간 IL-17A-PE가 포함된 1X permeabilization 용액과 빛 차단 하에 4℃에서 30분 동안 반응시켰다. FACS 완충액으로 세척한 후, 세포를 FACS 완충액에 재현탁시키고 유세포 분석기로 세포를 분석하였다.Collected mouse or human Th17 cells were placed in an ep tube, centrifuged, and washed with PBS. After washing, single cells were stimulated with 1X stimulation cocktail and 1X monensin at 37°C for 4 hours. Stimulated cells were centrifuged and reacted with anti-mouse or human CD4-PEcy7 staining buffer at 4° C. for 30 minutes under light blocking. After washing, a fixation/permeabilization solution was added to the sample and reacted at 4° C. for 18 hours under light blocking. After washing, the cells were reacted with 1X permeabilization solution containing anti-mouse or human IL-17A-PE at 4° C. for 30 minutes under light blocking. After washing with FACS buffer, the cells were resuspended in FACS buffer and analyzed by flow cytometry.
실시예 10: 정량적 실시간 중합효소 연쇄반응(qRT-PCR) 분석Example 10: Quantitative real-time polymerase chain reaction (qRT-PCR) analysis
qRT-PCR은 ABI StepOnePlus Real-Time PCR 시스템(Applied Biosystems, US)에서 SYBR® Green 실시간 PCR 키트(Mbiotech, Korea)를 사용하여 수행하였다. 유전자의 발현 수준은 GAPDH로 정규화하여 분석하였다.qRT-PCR was performed using the SYBR® Green real-time PCR kit (Mbiotech, Korea) in the ABI StepOnePlus Real-Time PCR system (Applied Biosystems, US). The expression level of the gene was analyzed by normalizing to GAPDH.
실시예 11: 세포 배양Example 11: Cell culture
MH7A 인간 류마티스 관절염 활막세포는 Cho YY 교수(Catholic University, Korea)로부터 제공받았다. MH7A 세포는 10% FBS, 페니실린(최종 농도, 100 U/ml), 스트렙토마이신(최종 농도, 0.1 mg/ml) 및 L-글루타민이 포함된 RPMI 1640 배지(Corning)를 이용하여 37℃, 5% C02 배양기에서 배양하였다.MH7A human rheumatoid arthritis synovial cells were provided by Professor Cho YY (Catholic University, Korea). MH7A cells were prepared using RPMI 1640 medium (Corning) containing 10% FBS, penicillin (final concentration, 100 U/ml), streptomycin (final concentration, 0.1 mg/ml) and L-glutamine at 37° C., 5%. Incubated in a CO 2 incubator.
LM 세포는 마우스 섬유아세포(fibroblast)로 Kim TS 교수(Korea university, Korea)로부터 제공받았다. LM 세포는 10% FBS, 페니실린(최종 농도, 100 U/ml), 스트렙토마이신(최종 농도, 0.1 mg/ml)이 포함된 DMEM 배지(Welgene)를 이용하여 37℃, 5% CO2 배양기에서 배양하였다.LM cells were mouse fibroblasts, which were provided by Professor Kim TS (Korea University, Korea). LM cells were cultured in a DMEM medium (Welgene) containing 10% FBS, penicillin (final concentration, 100 U/ml), and streptomycin (final concentration, 0.1 mg/ml) in a 37°C, 5% CO 2 incubator. I did.
실시예 12: 세포 생존율 분석Example 12: Cell viability analysis
세포 증식을 측정하기 위해, 세포(1 X 104 세포/200 μl/웰)를 96 웰 플레이트에 접종하고, 일반 배지를 첨가하여 72시간 동안 배양하였다. 이후, tetrazolim salt WST-1[2-(4-Iodophenyl)-3-(4-nitrophenyl)5-(2,4-disulfophenyl)-2H-tetrazolium]-based EZ-cytox 용액(Daeillab, Korea)을 사용하여 제조사에서 제공한 프로토콜에 따라 450 nm에서 흡광도를 측정하였다. 간략하게, 20 μl의 EZ- cytox 용액을 각 웰에 첨가하고, 37℃, 5% C02 배양기에서 1시간 동안 배양한 후, 450 nm에서 즉시 흡광도를 측정하였다.In order to measure cell proliferation, cells (1
실시예 13: TNF 중화 활성 분석Example 13: TNF neutralizing activity assay
TNF에 대한 독성 중화 활성을 측정하기 위해, 세포(5 X 104세포/100 μl/웰)를 96 웰 플레이트에 접종하고, 일반 배지를 사용하여 Actinomycin-D(AD) 0.5 μg/ml, AD+TNF 20 ng/ml, AD+TNF+THP(200 μM부터 1 nM까지 1/2씩 희석)를 처리한 후, 37℃, 5% CO2 배양기에서 24시간 동안 배양하였다. 이후, thiazolyl blue tetrazolium bromide(MTT, 5 mg/ml, Sigma, US)를 각 웰 당 10 μl씩 첨가하고 4시간 동안 추가 배양하였다. 이후, 상층액을 제거하고 200 μl의 DMSO를 각 웰에 첨가한 후 570 nm에서 흡광도를 측정하였다. To measure the toxic neutralizing activity for TNF, cells (5
실험예 1: 천연 단일 화합물에서 TNF-TNF 수용체의 결합을 억제하는 소분자 화합물 검출Experimental Example 1: Detection of small molecule compounds that inhibit the binding of TNF-TNF receptors in a single natural compound
TNF-TNF 수용체 사이의 결합을 억제하는 소분자 화합물을 검출하기 위해, TNF 경쟁 분석으로 스크리닝을 수행하였다. 플레이트를 TNF로 코팅한 후 TNFR2-Fc(etanercept, Enbrel)를 라이브러리의 각 천연 소분자 화합물과 동시에 첨가하여 TNF에 대한 경쟁적 결합을 유도하고, 대조군과 비교하여 OD 감소를 통해 TNF와 결합된 TNFR2를 검출하였다. TNF 경쟁 분석을 이용하여, 천연 단일 화합물 라이브러리(502 유형)를 스크리닝하였다. 도 1A와 같이, 두 가지 스크리닝 과정을 통해 TNF 표적 소분자 화합물인 THP를 동정하였으며, THP의 화학 구조는 도 1B에 나타내었다.In order to detect small molecule compounds that inhibit the binding between TNF-TNF receptors, screening was performed by TNF competition assay. After coating the plate with TNF, TNFR2-Fc (etanercept, Enbrel) was added simultaneously with each natural small molecule compound in the library to induce competitive binding to TNF, and compared with the control group, TNFR2 bound to TNF was detected through OD reduction. I did. Using the TNF competition assay, a native single compound library (502 type) was screened. As shown in FIG. 1A, THP, which is a TNF target small molecule compound, was identified through two screening processes, and the chemical structure of THP is shown in FIG. 1B.
결합 동역학을 측정하기 위해, biacore T200을 사용하여 SPR 분석을 수행하였다. 도 1C와 같이, THP는 CM5 칩에 고정화된 TNF에 대해 농도 의존적이고 특이적으로 포화 결합된 센서그램을 나타내었고, 결합 친화도(KD) 값은 1.663E-5였다. THP의 TNF 결합이 TNF의 시험관 내 세포독성을 억제할 수 있는지 여부를 평가하였다. 악티노마이신 D, TNF 및 THP를 마우스 섬유아세포인 LM 세포주에 처리하여 세포 독성을 유도하였다. 그 결과, 도 1D와 같이, THP는 농도 의존적으로 TNF의 세포독성을 차단하고, THP의 IC50 값은 0.4 μM임을 확인하였다.To measure binding kinetics, SPR analysis was performed using biacore T200. As shown in FIG. 1C, THP showed a concentration-dependent and specifically saturated-bound sensorgram for TNF immobilized on the CM5 chip, and the binding affinity (KD) value was 1.663E -5 . It was evaluated whether the TNF binding of THP could inhibit the cytotoxicity of TNF in vitro. Cytotoxicity was induced by treatment with actinomycin D, TNF, and THP on the LM cell line, which is a mouse fibroblast. As a result, as shown in FIG. 1D, it was confirmed that THP blocked the cytotoxicity of TNF in a concentration-dependent manner, and the IC50 value of THP was 0.4 μM.
실험예 2: CIA 모델에서 THP에 의한 증상 억제 효과Experimental Example 2: Effect of suppressing symptoms by THP in CIA model
CIA 동물 모델에서 THP의 치료 효과를 평가하기 위해, 2차 면역 후 비히클 또는 THP(5 mg/kg)를 매일 경구 투여하였다. 그 결과, 도 2A와 같이, 비히클이 투여된 군에 비해 THP(5 mg/kg)가 투여된 군에서 관절염 점수가 유의하게 감소하는 것을 확인하였다.To evaluate the therapeutic effect of THP in the CIA animal model, vehicle or THP (5 mg/kg) was administered orally daily after the second immunization. As a result, as shown in FIG. 2A, it was confirmed that the arthritis score significantly decreased in the group administered with THP (5 mg/kg) compared to the group administered with the vehicle.
다음으로, 비히클 또는 THP가 투여된 마우스의 혈청을 이용하여 염증성 사이토카인 분비에 대한 THP의 효능을 평가하였다. 그 결과, 도 2B와 같이, 비히클이 투여된 군에 비해 THP가 투여된 군에서 TNF, IL-6 및 IL-1β의 수준이 유의하게 감소하는 것을 확인하였다. 또한, 뒷다리 관절을 헤마톡실린 및 에오신으로 염색하여 조직학적 분석을 수행한 결과, 도 2C(상부)와 같이, THP가 투여된 군에서 염증, 연골 손상 및 뼈의 침식이 약화되는 것을 확인하였고, 사프라닌-O로 염색한 결과, 도 2C(하부)와 같이, 연골 손실이 방지되는 것을 확인할 수 있었다.Next, the efficacy of THP on the secretion of inflammatory cytokines was evaluated using the vehicle or serum of mice administered THP. As a result, as shown in FIG. 2B, it was confirmed that the levels of TNF, IL-6, and IL-1β were significantly reduced in the THP-administered group compared to the vehicle-administered group. In addition, as a result of performing histological analysis by staining the hind limb joints with hematoxylin and eosin, it was confirmed that inflammation, cartilage damage, and bone erosion were weakened in the THP-administered group, as shown in FIG. 2C (top), As a result of staining with safranin-O, it was confirmed that cartilage loss was prevented, as shown in FIG. 2C (lower part).
실험예 3: THP에 의한 마우스 및 인간 Th17 세포의 분화 및 기능 억제 효과Experimental Example 3: Effect of THP on differentiation and function inhibition of mouse and human Th17 cells
분화/활성화된 Th17 세포는 염증성 사이토카인인 IL-17A를 분비하고, 분비된 IL-17A는 관절 조직의 활막 섬유아세포에 영향을 미쳐 염증 반응을 유발하는 것으로 알려져 있다. Differentiated/activated Th17 cells secrete IL-17A, an inflammatory cytokine, and secreted IL-17A is known to affect synovial fibroblasts in joint tissues and induce an inflammatory response.
이에, 마우스 비장세포 및 인간 PBMC를 사용한 Th17 세포 분화에서 THP의 효과를 평가하였다. 그 결과, 도 3A 및 도 3B와 같이, 마우스 비장세포에서, THP는 농도 의존적으로 Th17 세포의 분화를 억제하고, THP(50 μM)는 Th17 세포에 의해 분비된 IL-17A 단백질 및 mRNA의 발현을 감소시키는 것을 확인하였다. 또한, 도 3C와 같이, Th17 관련 분자인 RORγT의 mRNA 발현을 감소시키는 것을 확인하였다.Thus, the effect of THP on Th17 cell differentiation using mouse splenocytes and human PBMCs was evaluated. As a result, as shown in FIGS. 3A and 3B, in mouse splenocytes, THP inhibits the differentiation of Th17 cells in a concentration-dependent manner, and THP (50 μM) inhibits the expression of IL-17A protein and mRNA secreted by Th17 cells. It was confirmed to decrease. In addition, as shown in Fig. 3C, it was confirmed that the mRNA expression of RORγT, a Th17-related molecule, was reduced.
마찬가지로, 도 4A 및 도 4B와 같이, 인간 PBMC에서, THP(12.5, 25 및 50 μM)는 Th17 세포의 분화를 유의하게 억제하고, 농도 의존적으로 Th17 세포에 의해 분비된 IL-17 단백질 및 mRNA의 발현을 유의하게 감소시키는 것을 확인하였다. 또한, 도 4C와 같이, THP(12.5 μM)는 RORγT의 mRNA 발현을 감소시키는 것을 확인하였다.Similarly, as shown in Figs. 4A and 4B, in human PBMCs, THP (12.5, 25 and 50 μM) significantly inhibits the differentiation of Th17 cells, and concentration-dependently of IL-17 proteins and mRNA secreted by Th17 cells. It was confirmed that the expression was significantly reduced. In addition, as shown in Figure 4C, it was confirmed that THP (12.5 μM) reduced the mRNA expression of RORγT.
실험예 4: 류마티스 관절염의 섬유아세포 유사 활막세포에서 THP에 의한 세포 증식 억제 효과Experimental Example 4: Inhibitory effect of THP on cell proliferation in fibroblast-like synovial cells of rheumatoid arthritis
THP가 류마티스 관절염의 섬유아세포 유사 활막세포(fibroblast like synoviocyte; FLS)의 증식을 억제할 수 있는지 여부를 평가하기 위해, 인간 류마티스 관절염 환자의 FLS 세포주인 MH7A 세포를 사용하여 MTT 분석을 수행하였다. 그 결과, 도 5와 같이, THP가 농도 의존적으로 MH7A 세포의 증식을 유의하게 억제 하는 것을 확인하였다.To evaluate whether THP can inhibit the proliferation of fibroblast-like synoviocytes (FLS) of rheumatoid arthritis, MTT analysis was performed using MH7A cells, which are FLS cell lines of human rheumatoid arthritis patients. As a result, as shown in FIG. 5, it was confirmed that THP significantly inhibited the proliferation of MH7A cells in a concentration-dependent manner.
이상으로 본 발명의 특정한 부분을 상세히 기술한 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현 예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.As the specific parts of the present invention have been described in detail above, it is clear that these specific techniques are only preferred embodiments, and the scope of the present invention is not limited thereto for those of ordinary skill in the art. Accordingly, it will be said that the substantial scope of the present invention is defined by the appended claims and their equivalents.
본 발명의 범위는 후술하는 특허청구범위에 의하여 나타내어지며, 특허청구범위의 의미 및 범위 그리고 그 균등 개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범위에 포함되는 것으로 해석되어야 한다.The scope of the present invention is indicated by the claims to be described later, and all changes or modified forms derived from the meaning and scope of the claims and their equivalent concepts should be construed as being included in the scope of the present invention.
Claims (9)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020190132187A KR20210048208A (en) | 2019-10-23 | 2019-10-23 | Composition for preventing or treating TNF-mediated diseases comprising tetrahydropapaverine and method for inhibiting TNF-activity with the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020190132187A KR20210048208A (en) | 2019-10-23 | 2019-10-23 | Composition for preventing or treating TNF-mediated diseases comprising tetrahydropapaverine and method for inhibiting TNF-activity with the same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| KR20210048208A true KR20210048208A (en) | 2021-05-03 |
Family
ID=75910987
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020190132187A KR20210048208A (en) | 2019-10-23 | 2019-10-23 | Composition for preventing or treating TNF-mediated diseases comprising tetrahydropapaverine and method for inhibiting TNF-activity with the same |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR20210048208A (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20120093534A (en) | 2011-02-15 | 2012-08-23 | 재단법인 전라남도생물산업진흥재단 | Pharmaceutical composition for preventing and treating rheumatoid arthritis comprising extracts of leafs of eriobotrya japonica |
-
2019
- 2019-10-23 KR KR1020190132187A patent/KR20210048208A/en not_active Application Discontinuation
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20120093534A (en) | 2011-02-15 | 2012-08-23 | 재단법인 전라남도생물산업진흥재단 | Pharmaceutical composition for preventing and treating rheumatoid arthritis comprising extracts of leafs of eriobotrya japonica |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP2022092052A (en) | Composition for prevention or treatment of tnf-related diseases containing novel derivative as effective component and tnf activation inhibitor therewith | |
| CN103648502B (en) | Include the oleanolic acid acetic acid esters pharmaceutical composition disease mediated as the prevention of active ingredient or treatment TLR and IL 6 | |
| SA04240497A (en) | Low dose methods for treating disorders in which tnf c activity is detrimental | |
| JP2015518861A (en) | Pharmaceutical composition for preventing or treating STAT3-mediated diseases, comprising an extract of Mizokouju or a fraction thereof as an active ingredient | |
| EP2968437A1 (en) | Reducing proinflammatory response | |
| KR102150944B1 (en) | Composition for prevention and treatment of muscle strengthening or treating of sarcopenia | |
| KR20130107531A (en) | Pharmaceutical composition for prevention and treatment of chronic obstructive pulmonary disease comprising extract of lilium lancifolium thunberg as an active ingredient | |
| JP7060500B2 (en) | A method for producing a pharmaceutical composition, a method for producing a quasi-drug composition, a method for producing a feed composition, and a method for producing a drinking water additive. | |
| US20150182487A1 (en) | Composition for preventing and treating inflammatory diseases and immune diseases, containing apo-9`-fucoxanthinone as active ingredient | |
| KR102105483B1 (en) | Composition for preventing or treating TNF-mediated diseases comprising tetrahydropapaverine and IL-6 inhibitor, and method for inhibiting TNF-activity with the same | |
| TWI504394B (en) | Method for preparing antofine | |
| US20160074338A1 (en) | Food composition for preventing obesity, pharmaceutical composition for treating obesity, and animal medicine for treating obesity, containing gingernone a | |
| US9700531B2 (en) | Composition comprising metformin as active ingredient for preventing or treating inflammatory bowel disease | |
| KR20130117491A (en) | Composition for preventing and treating inflammatory or immune diseases comprising plocamium telfairiae extract | |
| KR101673265B1 (en) | A pharmaceutical composition for preventing or treating IL-6-mediated disease, comprising extract, fraction, or compounds derived from Ampelopsis brevipedunculata | |
| KR20210048208A (en) | Composition for preventing or treating TNF-mediated diseases comprising tetrahydropapaverine and method for inhibiting TNF-activity with the same | |
| JP2021523952A (en) | A composition for the prevention or treatment of allergic diseases containing an inotodiol compound as an active ingredient. | |
| KR101735061B1 (en) | Composition containing Artemisia annua extract, artemisinin or dihydroartemisinin for preventing or treating obesity | |
| KR101354167B1 (en) | Novel compound of Ohionsin F for Prevention and Treatment of Atherosclerosis from Polytrichastrum alpinum | |
| KR20220044057A (en) | Fusion protein comprising Interleukin-18 binding protein and antigen binding fragment to serum albumin, and uses thereof | |
| KR101184343B1 (en) | Composition for preventing and treating inflammatory disease comprising piperine or pharmaceutically acceptable salt thereof as an active ingredient | |
| KR101892577B1 (en) | Composition for preventing or treating atherosclerosis comprising reblastatin derivation or pharmaceutical acceptable salts thereof | |
| KR20170092490A (en) | Pharmaceutical composition for prevention or treatment of immune diseases comprising 4-hydroxy-2,3-dimethyl-2-nonen-4-olide | |
| KR101273953B1 (en) | Novel compound of Sterocalpin A for Prevention and Treatment of Atherosclerosis | |
| KR101777497B1 (en) | Pharmaceutical composition for prevention or treatment of immune diseases comprising 4-hydroxy-2,3-dimethyl-2-nonen-4-olide or 3-hydroxy-4,7-megastigmadien-9-one compounds |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| E902 | Notification of reason for refusal | ||
| E601 | Decision to refuse application |