CN117503785A - Application of larch resin alcohol and derivative thereof in preparation of COX-2 selective inhibitor - Google Patents
Application of larch resin alcohol and derivative thereof in preparation of COX-2 selective inhibitor Download PDFInfo
- Publication number
- CN117503785A CN117503785A CN202311637681.8A CN202311637681A CN117503785A CN 117503785 A CN117503785 A CN 117503785A CN 202311637681 A CN202311637681 A CN 202311637681A CN 117503785 A CN117503785 A CN 117503785A
- Authority
- CN
- China
- Prior art keywords
- cox
- resin alcohol
- larch
- larch resin
- alcohol
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 title claims abstract description 111
- 239000011347 resin Substances 0.000 title claims abstract description 100
- 229920005989 resin Polymers 0.000 title claims abstract description 100
- 241000218652 Larix Species 0.000 title claims abstract description 97
- 235000005590 Larix decidua Nutrition 0.000 title claims abstract description 97
- 229940111134 coxibs Drugs 0.000 title claims abstract description 27
- 238000002360 preparation method Methods 0.000 title claims description 24
- 150000002148 esters Chemical class 0.000 claims abstract description 4
- 150000003839 salts Chemical class 0.000 claims abstract description 4
- 230000003110 anti-inflammatory effect Effects 0.000 claims abstract description 3
- 230000001754 anti-pyretic effect Effects 0.000 claims abstract description 3
- 239000002221 antipyretic Substances 0.000 claims abstract description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 238000002156 mixing Methods 0.000 claims description 9
- -1 aqua Substances 0.000 claims description 8
- 239000010231 banlangen Substances 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 7
- 238000002347 injection Methods 0.000 claims description 7
- 239000007924 injection Substances 0.000 claims description 7
- 239000003826 tablet Substances 0.000 claims description 7
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 6
- 239000000284 extract Substances 0.000 claims description 6
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 6
- 238000011068 loading method Methods 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 238000005303 weighing Methods 0.000 claims description 6
- 150000001298 alcohols Chemical class 0.000 claims description 4
- 238000000605 extraction Methods 0.000 claims description 4
- 239000008187 granular material Substances 0.000 claims description 4
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 4
- 238000004440 column chromatography Methods 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 239000003937 drug carrier Substances 0.000 claims description 3
- 239000000706 filtrate Substances 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 3
- 125000003158 alcohol group Chemical group 0.000 claims description 2
- 229940035676 analgesics Drugs 0.000 claims description 2
- 239000000730 antalgic agent Substances 0.000 claims description 2
- 229940124599 anti-inflammatory drug Drugs 0.000 claims description 2
- 239000002246 antineoplastic agent Substances 0.000 claims description 2
- 229940041181 antineoplastic drug Drugs 0.000 claims description 2
- 229940046011 buccal tablet Drugs 0.000 claims description 2
- 239000006189 buccal tablet Substances 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 239000002502 liposome Substances 0.000 claims description 2
- 239000008176 lyophilized powder Substances 0.000 claims description 2
- 239000004005 microsphere Substances 0.000 claims description 2
- 239000006072 paste Substances 0.000 claims description 2
- 229940023488 pill Drugs 0.000 claims description 2
- 239000006187 pill Substances 0.000 claims description 2
- 239000007901 soft capsule Substances 0.000 claims description 2
- 239000007921 spray Substances 0.000 claims description 2
- 239000006190 sub-lingual tablet Substances 0.000 claims description 2
- 229940098466 sublingual tablet Drugs 0.000 claims description 2
- 238000001179 sorption measurement Methods 0.000 claims 2
- 230000000202 analgesic effect Effects 0.000 claims 1
- 102100038280 Prostaglandin G/H synthase 2 Human genes 0.000 abstract description 30
- 230000000694 effects Effects 0.000 abstract description 17
- 239000003255 cyclooxygenase 2 inhibitor Substances 0.000 abstract description 8
- 239000000463 material Substances 0.000 abstract description 5
- 241000196324 Embryophyta Species 0.000 abstract description 3
- 102000004005 Prostaglandin-endoperoxide synthases Human genes 0.000 abstract description 3
- 108090000459 Prostaglandin-endoperoxide synthases Proteins 0.000 abstract description 3
- 231100000331 toxic Toxicity 0.000 abstract description 3
- 230000002588 toxic effect Effects 0.000 abstract description 3
- 206010067484 Adverse reaction Diseases 0.000 abstract description 2
- 230000006838 adverse reaction Effects 0.000 abstract description 2
- 108050003267 Prostaglandin G/H synthase 2 Proteins 0.000 abstract 2
- 239000004480 active ingredient Substances 0.000 abstract 1
- 230000001760 anti-analgesic effect Effects 0.000 abstract 1
- 230000000259 anti-tumor effect Effects 0.000 abstract 1
- 108010037462 Cyclooxygenase 2 Proteins 0.000 description 29
- 230000005764 inhibitory process Effects 0.000 description 29
- 210000004027 cell Anatomy 0.000 description 22
- 108010037464 Cyclooxygenase 1 Proteins 0.000 description 21
- 102100038277 Prostaglandin G/H synthase 1 Human genes 0.000 description 21
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 18
- 239000003814 drug Substances 0.000 description 18
- 229960000590 celecoxib Drugs 0.000 description 15
- RZEKVGVHFLEQIL-UHFFFAOYSA-N celecoxib Chemical compound C1=CC(C)=CC=C1C1=CC(C(F)(F)F)=NN1C1=CC=C(S(N)(=O)=O)C=C1 RZEKVGVHFLEQIL-UHFFFAOYSA-N 0.000 description 15
- 229940079593 drug Drugs 0.000 description 14
- BSYNRYMUTXBXSQ-UHFFFAOYSA-N Aspirin Chemical compound CC(=O)OC1=CC=CC=C1C(O)=O BSYNRYMUTXBXSQ-UHFFFAOYSA-N 0.000 description 11
- 239000000243 solution Substances 0.000 description 11
- 229960001138 acetylsalicylic acid Drugs 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 9
- 238000000034 method Methods 0.000 description 9
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 8
- 102000004190 Enzymes Human genes 0.000 description 8
- 108090000790 Enzymes Proteins 0.000 description 8
- 229940088598 enzyme Drugs 0.000 description 8
- 229960000371 rofecoxib Drugs 0.000 description 8
- RZJQGNCSTQAWON-UHFFFAOYSA-N rofecoxib Chemical compound C1=CC(S(=O)(=O)C)=CC=C1C1=C(C=2C=CC=CC=2)C(=O)OC1 RZJQGNCSTQAWON-UHFFFAOYSA-N 0.000 description 8
- 238000012360 testing method Methods 0.000 description 8
- 229940021182 non-steroidal anti-inflammatory drug Drugs 0.000 description 7
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 6
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 6
- 230000009471 action Effects 0.000 description 6
- 239000012091 fetal bovine serum Substances 0.000 description 6
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 6
- MHXCIKYXNYCMHY-AUSJPIAWSA-N (+)-lariciresinol Chemical compound C1=C(O)C(OC)=CC(C[C@@H]2[C@@H]([C@H](OC2)C=2C=C(OC)C(O)=CC=2)CO)=C1 MHXCIKYXNYCMHY-AUSJPIAWSA-N 0.000 description 5
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 5
- YVRYZXAHRGGELT-UHFFFAOYSA-N Lariciresinol Natural products C1=C2OCOC2=CC(C2C(C)C3(OC)C=C(CC=C)C(=O)CC3(O2)OC)=C1 YVRYZXAHRGGELT-UHFFFAOYSA-N 0.000 description 5
- 230000002411 adverse Effects 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 150000002191 fatty alcohols Chemical class 0.000 description 5
- 230000002496 gastric effect Effects 0.000 description 5
- 239000003112 inhibitor Substances 0.000 description 5
- 235000006826 lariciresinol Nutrition 0.000 description 5
- 229940094443 oxytocics prostaglandins Drugs 0.000 description 5
- 150000003180 prostaglandins Chemical class 0.000 description 5
- 238000012216 screening Methods 0.000 description 5
- DZUXGQBLFALXCR-UHFFFAOYSA-N (+)-(9alpha,11alpha,13E,15S)-9,11,15-trihydroxyprost-13-en-1-oic acid Natural products CCCCCC(O)C=CC1C(O)CC(O)C1CCCCCCC(O)=O DZUXGQBLFALXCR-UHFFFAOYSA-N 0.000 description 4
- OKKJLVBELUTLKV-MZCSYVLQSA-N Deuterated methanol Chemical compound [2H]OC([2H])([2H])[2H] OKKJLVBELUTLKV-MZCSYVLQSA-N 0.000 description 4
- 229910002092 carbon dioxide Inorganic materials 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 230000002526 effect on cardiovascular system Effects 0.000 description 4
- 229930013686 lignan Natural products 0.000 description 4
- 235000009408 lignans Nutrition 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000001543 one-way ANOVA Methods 0.000 description 4
- DZUXGQBLFALXCR-CDIPTNKSSA-N prostaglandin F1alpha Chemical compound CCCCC[C@H](O)\C=C\[C@H]1[C@H](O)C[C@H](O)[C@@H]1CCCCCCC(O)=O DZUXGQBLFALXCR-CDIPTNKSSA-N 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
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 3
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 3
- 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 3
- 239000004471 Glycine Substances 0.000 description 3
- 206010061218 Inflammation Diseases 0.000 description 3
- 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 3
- 229930195725 Mannitol Natural products 0.000 description 3
- 229920000881 Modified starch Polymers 0.000 description 3
- CMWTZPSULFXXJA-UHFFFAOYSA-N Naproxen Natural products C1=C(C(C)C(O)=O)C=CC2=CC(OC)=CC=C21 CMWTZPSULFXXJA-UHFFFAOYSA-N 0.000 description 3
- 208000002193 Pain Diseases 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 210000001035 gastrointestinal tract Anatomy 0.000 description 3
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 3
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 3
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 3
- UFVKGYZPFZQRLF-UHFFFAOYSA-N hydroxypropyl methyl cellulose Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 UFVKGYZPFZQRLF-UHFFFAOYSA-N 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 230000004054 inflammatory process Effects 0.000 description 3
- 239000008101 lactose Substances 0.000 description 3
- GAYKAIAESJROGN-UHFFFAOYSA-N lariciresinol-4-O-beta-D-glucoside Natural products C1=C(O)C(OC)=CC(C2C(C(CC=3C=C(OC)C(OC4C(C(O)C(O)C(CO)O4)O)=CC=3)CO2)CO)=C1 GAYKAIAESJROGN-UHFFFAOYSA-N 0.000 description 3
- 235000019359 magnesium stearate Nutrition 0.000 description 3
- 239000000594 mannitol Substances 0.000 description 3
- 235000010355 mannitol Nutrition 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 229960002009 naproxen Drugs 0.000 description 3
- CMWTZPSULFXXJA-VIFPVBQESA-N naproxen Chemical compound C1=C([C@H](C)C(O)=O)C=CC2=CC(OC)=CC=C21 CMWTZPSULFXXJA-VIFPVBQESA-N 0.000 description 3
- 206010039073 rheumatoid arthritis Diseases 0.000 description 3
- 238000007873 sieving Methods 0.000 description 3
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- 235000002639 sodium chloride Nutrition 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 239000008215 water for injection Substances 0.000 description 3
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- 101100275473 Caenorhabditis elegans ctc-3 gene Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 241000334160 Isatis Species 0.000 description 2
- 241000334154 Isatis tinctoria Species 0.000 description 2
- 101100168274 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) cox-3 gene Proteins 0.000 description 2
- 108010019160 Pancreatin Proteins 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
- 206010037660 Pyrexia Diseases 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000001569 carbon dioxide Substances 0.000 description 2
- 239000006143 cell culture medium Substances 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000007405 data analysis Methods 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- XEYBRNLFEZDVAW-ARSRFYASSA-N dinoprostone Chemical compound CCCCC[C@H](O)\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1C\C=C/CCCC(O)=O XEYBRNLFEZDVAW-ARSRFYASSA-N 0.000 description 2
- 229960002986 dinoprostone Drugs 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 150000005692 lignans Chemical class 0.000 description 2
- 210000002540 macrophage Anatomy 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 201000008482 osteoarthritis Diseases 0.000 description 2
- 229940055695 pancreatin Drugs 0.000 description 2
- 229940049954 penicillin Drugs 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- 239000000419 plant extract Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- XEYBRNLFEZDVAW-UHFFFAOYSA-N prostaglandin E2 Natural products CCCCCC(O)C=CC1C(O)CC(=O)C1CC=CCCCC(O)=O XEYBRNLFEZDVAW-UHFFFAOYSA-N 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000012453 solvate Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000000699 topical effect Effects 0.000 description 2
- 239000012588 trypsin Substances 0.000 description 2
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 201000004569 Blindness Diseases 0.000 description 1
- 241000219193 Brassicaceae Species 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 206010059024 Gastrointestinal toxicity Diseases 0.000 description 1
- HEFNNWSXXWATRW-UHFFFAOYSA-N Ibuprofen Chemical compound CC(C)CC1=CC=C(C(C)C(O)=O)C=C1 HEFNNWSXXWATRW-UHFFFAOYSA-N 0.000 description 1
- 108010044467 Isoenzymes Proteins 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 206010068319 Oropharyngeal pain Diseases 0.000 description 1
- 201000007100 Pharyngitis Diseases 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 235000009697 arginine Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- CCGSUNCLSOWKJO-UHFFFAOYSA-N cimetidine Chemical compound N#CNC(=N/C)\NCCSCC1=NC=N[C]1C CCGSUNCLSOWKJO-UHFFFAOYSA-N 0.000 description 1
- 229960001380 cimetidine Drugs 0.000 description 1
- 229940075614 colloidal silicon dioxide Drugs 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 208000000718 duodenal ulcer Diseases 0.000 description 1
- 230000002900 effect on cell Effects 0.000 description 1
- KAQKFAOMNZTLHT-VVUHWYTRSA-N epoprostenol Chemical compound O1C(=CCCCC(O)=O)C[C@@H]2[C@@H](/C=C/[C@@H](O)CCCCC)[C@H](O)C[C@@H]21 KAQKFAOMNZTLHT-VVUHWYTRSA-N 0.000 description 1
- 229960001123 epoprostenol Drugs 0.000 description 1
- BEFDCLMNVWHSGT-UHFFFAOYSA-N ethenylcyclopentane Chemical compound C=CC1CCCC1 BEFDCLMNVWHSGT-UHFFFAOYSA-N 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 231100000414 gastrointestinal toxicity Toxicity 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 239000003485 histamine H2 receptor antagonist Substances 0.000 description 1
- 229960001680 ibuprofen Drugs 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 150000002611 lead compounds Chemical class 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- OJLOPKGSLYJEMD-URPKTTJQSA-N methyl 7-[(1r,2r,3r)-3-hydroxy-2-[(1e)-4-hydroxy-4-methyloct-1-en-1-yl]-5-oxocyclopentyl]heptanoate Chemical compound CCCCC(C)(O)C\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1CCCCCCC(=O)OC OJLOPKGSLYJEMD-URPKTTJQSA-N 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229960005249 misoprostol Drugs 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 208000010125 myocardial infarction Diseases 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 150000003815 prostacyclins Chemical class 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 229940126409 proton pump inhibitor Drugs 0.000 description 1
- 239000000612 proton pump inhibitor Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007779 soft material Substances 0.000 description 1
- 239000004334 sorbic acid Substances 0.000 description 1
- 229940075582 sorbic acid Drugs 0.000 description 1
- 235000010199 sorbic acid Nutrition 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 238000000859 sublimation Methods 0.000 description 1
- 230000008022 sublimation Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 230000001732 thrombotic effect Effects 0.000 description 1
- RZWIIPASKMUIAC-VQTJNVASSA-N thromboxane Chemical compound CCCCCCCC[C@H]1OCCC[C@@H]1CCCCCCC RZWIIPASKMUIAC-VQTJNVASSA-N 0.000 description 1
- 229940124279 traditional non-steroidal anti-inflammatory drug Drugs 0.000 description 1
- 231100000397 ulcer Toxicity 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
-
- 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
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
- A61K31/341—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide not condensed with another ring, e.g. ranitidine, furosemide, bufetolol, muscarine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/19—Acanthaceae (Acanthus family)
- A61K36/195—Strobilanthes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/31—Brassicaceae or Cruciferae (Mustard family), e.g. broccoli, cabbage or kohlrabi
- A61K36/315—Isatis, e.g. Dyer's woad
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
- C07D307/02—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings
- C07D307/04—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having no double bonds between ring members or between ring members and non-ring members
- C07D307/10—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having no double bonds between ring members or between ring members and non-ring members with substituted hydrocarbon radicals attached to ring carbon atoms
- C07D307/12—Radicals substituted by oxygen atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/26—Acyclic or carbocyclic radicals, substituted by hetero rings
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Epidemiology (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Alternative & Traditional Medicine (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Rheumatology (AREA)
- Pain & Pain Management (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to application of larch resin alcohol and derivatives thereof in preparing COX-2 selective inhibitors, in particular to application of COX-2, wherein the target point is cyclooxygenase COX-2, the COX-2 inhibitors are larch resin alcohol, larch resin alcohol 4-O-beta-D-glucopyranoside and/or pharmaceutically acceptable polyglycoside, ester, salt and the like, and the larch resin alcohol, the larch resin alcohol 4-O-beta-D-glucopyranoside are derived from traditional Chinese medicinal materials or natural plants, and no obvious toxic or side effect or adverse reaction is found until now, so that the active ingredients are used as selective COX-2 inhibitors and have the relative antipyretic, anti-inflammatory, analgesic and antitumor medicinal application.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to application of larch resin alcohol and derivatives thereof in preparation of COX-2 selective inhibitors.
Background
Cyclooxygenase (COX) is an important rate-limiting enzyme that controls the production of Prostaglandins (PGs) and thromboxane (TxA 2). The enzyme has 3 isoenzymes: COX-1, COX-2 and COX-3.COX-1 catalyzes the production of PGs and TxA2, has the ability to regulate gastrointestinal, renal, vascular and other physiological functions, and COX-2 regulates the production of PGs involved in inflammation, pain and fever, COX-3 being in fact a splice variant of COX-1. At present, clinically used medicines are designed mainly for two targets of COX-1 and COX-2.
Traditional nonsteroidal anti-inflammatory drugs (NSAIDs) have poor selectivity for COX-1 and COX-2, whereas prostaglandins produced by COX-1 metabolism have gastric mucosal protective effects, which are blocked, causing serious gastrointestinal side effects. In order to reduce the side effects of the gastrointestinal tract, various measures have been taken, such as the use of the H2 receptor antagonist cimetidine to reduce the formation of duodenal ulcers, but most patients are not as effective. A more effective approach is to employ gastrointestinal tract protecting drugs such as proton pump inhibitors or the prostacyclin analogue misoprostol. Although this approach may reduce gastric lesions, it does not completely eliminate ulcer formation. NO release has also been used in combination with non-steroidal anti-inflammatory drugs. However, none of the above methods truly addresses the side effects caused by insufficient topical mucosal prostacyclin.
The selective COX-2 inhibitor is a novel NSAIDs, can selectively inhibit cell COX-2 causing pain and inflammation, does not interfere with COX-1 to protect stomach and intestinal linings, and can reduce adverse events of gastrointestinal complications caused by NSAIDs. However, the widely used COX-2 inhibitor rofecoxib causes an increasing report of cardiovascular adverse events, reicin [ Reicin A S, shapiro D, sprling R S, et al Comparison of cardiovascular thrombotic events in patients with osteoarthritis treated with Rofecoxib versus nonselective nonsteroidal anti-inflammatory drugs (Ibuprofen, dicrofenac, and N abumetone) [ J ]. American Journal of Cardiology,2002,89 (2): 204-209.] for 3 years of rofecoxib and 5435 patients in the clinical research project of osteoarthritis participate in double blindness tests, and the results show that both the administration of rofecoxib and non-selective NSAIDs can increase the incidence of thrombotic cardiovascular adverse events. Subsequently, bombardier et al [ Bombardier C, lane L, reicin A, et al Comparison of upper gastrointestinal toxicity of rofecoxib and naproxen in patients with rheumatoid arthritis [ J ]. New England Journal of Medicine,2000,343 (21): 1520-1528 ] randomly distributed 8076 cases of rheumatoid arthritis in 2 groups, and compared the incidence of adverse events in patients taking rofecoxib and naproxen, the results showed similar efficacy of rofecoxib and naproxen on rheumatoid arthritis, the latter causing 2 times the incidence of adverse events in the gastrointestinal tract, but 4 times the incidence of myocardial infarction. This series of results has forced the merck pharmaceutical company to announce the active withdrawal of the drug rofecoxib worldwide, and there is a clinical need for a safe and effective COX-2 inhibitor. Thus, chemically synthesized selective COX-2 inhibitors have potential cardiovascular risks.
Radix Isatidis is the dry root of Isatis tinctoria (Isatis indigotica Fort) of Brassicaceae, and has effects of clearing heat and toxic materials, cooling blood, and relieving sore throat. The isatis root has long history of application in China and high safety, and has no related report of toxic and side effects so far. The larch resin alcohol and the larch resin alcohol 4-O-beta-D-glucopyranoside are chemical components extracted and separated from the isatis root, and although relevant research reports on antiviral are provided, the contents of the larch resin alcohol and the larch resin alcohol 4-O-beta-D-glucopyranoside acting on COX-2 targets and relevant pharmaceutical uses are not reported.
Disclosure of Invention
The invention aims to provide application of larch resin alcohol and derivatives thereof in preparing COX-2 selective inhibitors.
The invention relates to the discovery of an action target point of a lignan compound derivative from a natural plant source, in particular to the discovery of an action target point of a larch resin alcohol, a larch She Songshu resin alcohol 4-O-beta-D-glucopyranoside and a larch resin alcohol-containing 4-O-beta-D-glucopyranoside plant extract contained in the lignan compound, wherein the action target point is cyclooxygenase COX-2, and the pharmaceutical application related to COX-2 possibly is the application of a medicament for relieving fever, resisting inflammation and easing pain related to COX-2 inhibition.
The application of larch resin alcohol and derivatives thereof in preparing COX-2 selective inhibitors, wherein the structural formula of the larch resin alcohol is I, one of the larch resin alcohol derivatives is larch She Songshu resin alcohol 4-O-beta-D-glucopyranoside, the structural formula is II,
the use of larch resin alcohol and derivatives thereof in the preparation of COX-2 selective inhibitors, the larch resin alcohol derivatives also including polyglycosides, esters, salts, solvates, plant extracts, chemical compositions, and pharmaceutical compositions containing larch resin alcohol groups.
The application of the larch resin alcohol and the derivative thereof in preparing COX-2 selective inhibitors comprises two or more of larch resin alcohol, larch resin alcohol 4-O-beta-D-glucopyranoside, polyglycoside, ester, salt and solvate, or pharmaceutically acceptable carriers or excipients.
The COX-2 selective inhibitor is in the form of injection, lyophilized powder for injection, injectable microsphere, liposome, bilayer/multilayer tablet, buccal tablet, sublingual tablet, capsule, aqua, powder, paste, spray, granule, soft capsule, dripping pill, gel, patch, or paste.
The application of the larch resin alcohol and the derivative thereof in preparing COX-2 selective inhibitors and the application of the COX-2 selective inhibitors in preparing antipyretic, anti-inflammatory, analgesic and antitumor drugs.
The pharmaceutically acceptable carrier or excipient is not limited to the choice of carrier or excipient, for example: starch, sodium chloride, microcrystalline cellulose, lactose, pregelatinized starch, sodium carboxymethyl cellulose, hydroxypropyl methylcellulose, magnesium stearate, colloidal silicon dioxide, glycine, arginine, sorbic acid, mannitol, and the like. The composition may be administered by, but is not limited to, intravenous, oral, intramuscular, subcutaneous, topical, and the like.
The beneficial effects are that:
the invention provides a safe natural plant-derived selective COX-2 inhibitor, which has selectivity on COX-2 inhibition and weak or no inhibition on COX-1, has similar drug effect to that of celecoxib which is clinically used at present, and is expected to avoid adverse reaction events of the drugs. The discovery of definite action targets and purposes of the components provides novel lead compounds for the discovery and application of potential novel non-steroidal anti-inflammatory drugs, and has definite development and application prospects.
Drawings
FIG. 1 is a graph showing the efficacy of lariciresinol, 4-O-beta-D-glucopyranoside, on COX-2 inhibition.
Figure 2 is a graph showing the efficacy of celecoxib as a control against COX-2 inhibition.
FIG. 3 is a graph showing COX-1 inhibition by lariciresinol, lariciresinol 4-O-beta-D-glucopyranoside. Wherein, the concentration of the larch resin alcohol, the larch resin alcohol 4-O-beta-D-glucopyranoside and the aspirin is 100 mu M, and the concentration of the celecoxib is 1 mu M.
FIG. 4 is the effect of test agents on RAW264.7 cell proliferation.
FIG. 5 shows COX-2 inhibition by test agents on LPS-induced RAW264.7 cell model.
FIG. 6 shows COX-1 inhibition in LPS-induced RAW264.7 cell model by test agents.
Detailed Description
The invention is further described below in conjunction with the detailed description. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. Further, it is understood that various changes and modifications of the present invention may be made by those skilled in the art after reading the description of the present invention, and that such equivalents are intended to fall within the scope of the claims appended hereto.
Example 1: extraction and separation of larch resin alcohol and larch resin alcohol 4-O-beta-D-glucopyranoside
1. The extraction method comprises the following steps: weighing 50 kg of radix isatidis decoction pieces, sequentially reflux-extracting with 8 times of 80% ethanol for 3 times each for 1 hour, filtering, mixing filtrates, recovering ethanol, and extracting with n-butanol to obtain extract. Loading into glass chromatographic column, adding 2 times of distilled water into the extract (1.8 Kg), suspending in an ultrasonic instrument, filtering, loading onto D101 macroporous adsorbent resin column, eluting sample with 10%, 20%, 30%, 40%, 50% and 60% ethanol sequentially, collecting fraction 500mL as one unit, concentrating, and drying to obtain total lignan sample of radix Isatidis total lignan sample. Separating the sample by octadecyl silane chemically bonded phase (C18) column chromatography, eluting with 7%, 20%, 30%, 40%, 50%, and pure methanol, detecting the sample by HPLC, and mixing the eluted samples according to HPLC profile data to obtain monomer compounds, namely larch resin alcohol and larch resin alcohol 4-O-beta-D-glucopyranoside.
2. Authentication method
Larch resin alcohol: white crystals. m/z 383[ M+Na ] +.
1H-NMR(400MHz,CD3OD),δ:6.83(H-2,s,1H),3.74(OCH 3 -3,s,3H),8.75(H-4,s,2H),6.75(H-5,d,J＝1.5Hz),6.67-6.71(H-6,2′,5′,overlap,3H),4.66(H-7,d,J＝6.4Hz,1H),2.19(H-8,m,1H),3.47(H-9a,dd,J＝6.8Hz,1H),3.65(H-9b,m,1H),3.73(OCH 3 -3′,s,3H),8.75(H-4′,s,2H),6.58(H-6′,dd,J＝1.6,6.4Hz,1H),2.42(H-7′a,dd,J＝13.5,11.0Hz,1H),2.83(H-7′b,dd,J＝13.5,11.0Hz,1H),2.53(H-8′,m,1H),3.55(H-9′a,dd,J＝7.0,7.5Hz,1H),3.88(H-9′b,dd,7.0,7.5Hz,1H)；
13C-NMR(100MHz,CD3OD),δ:135.17(C-1),110.42(C-2),147.83(OCH 3 -3),145.99(C-4),115.51(C-5),118.65(C-6),82.23(C-7),52.89(C-8),59.07(C-9),132.20(C-1′),113.18(C-2′),147.92(OCH 3 -3′),145.03(C-4′),115.85(C-5′),121.06(C-6′),32.62(C-7′),42.45(C-8′),72.28(C-9′)。
Fall She Songshu fatty alcohol 4-O-beta-D-glucopyranoside: white powder, m/z:523[ M+H ]] + 。
1 H-NMR(400MHz,DMSO),δ:8.683(-OH,1H,s),7.037、7.016(H-5,d,J＝8.4Hz),6.891(H-2,s,1H),6.783(H-6,dd,J＝1.6,8.4Hz),6.752(H-2′,d,J＝1.6Hz),6.684、6.664(d,J＝8.0Hz,H-5′),6.577(H-6′,dd,J＝1.6Hz,8.0Hz),4.70-4.729(H-7,H-1″,2H),3.455(br.d,J＝6.0Hz,H-9α),3.672(br.d,J＝7.2Hz,H-9β),2.199(m,H-8′),3.579(dd,J＝7.1,6.8Hz,H-9′α),3.89(dd,J＝7.1,6.4Hz,H-9′β),3.743(s,-OCH 3 ×2)；
13 C-NMR(100MHz,DMSO),δ:32.11(C-7′),42.0(C-8′),52.6(C-8),55.7(OCH 3 ),58.7(C-9),60.8(C-6″),69.8(C-4″),71.9(C-9′),73.3(C-2″),76.9(C-5″),77.1(C-3″),81.6(C-7),100.2(C-2),110.2(C-1″),112.8(C-2′),115.2(C-5),115.4(C-5′),117.9(C-6),120.6(C-6′),131.68(C-1′),137.66(C-1),144.56(C-4′),145.52(C-4),147.44(C-3′),148.78(C-3)。
Example 2: in vitro selective COX-2 direct inhibition of lariciresinol, lariciresinol 4-O-beta-D-glucopyranoside
1. Experimental materials: cyclooxygenase-1 (COX-1) inhibitor screening kit, available from Sigma Co., USA. Cyclooxygenase-2 (COX-2) inhibitor screening kit, available from Biyundian reagent company. Varioskan Flash multifunctional microplate reader, sieimer Feier technologies, USA.
2. Experimental procedure
2.1 preparation of control drug: 100 mu M celecoxib in a2 mu LCOX-2 inhibitor screening kit is taken, 198 mu L of DMSO is added, and the mixture is uniformly mixed to prepare 1 mu M solution, and aspirin is the 100 mu M solution of the kit.
2.2 preparation of test drug: 3.60mg of larch resin alcohol prepared in example 1 and 5.21mg of larch resin alcohol 4-O-beta-D-glucopyranoside were precisely weighed into 2 EP tubes, 50 mu of LDMSO was added thereto, and the mixture was dissolved by ultrasonic waves to prepare a 2mM solution. The gradient was diluted to 8 concentrations.
2.3 inhibition of COX-2 by lariciresinol, 4-O-beta-D-glucopyranoside
2.3.1 setting 3 multiple holes for each detection sample, strictly operating according to the instruction manual of the cyclooxygenase-2 (COX-2) inhibitor screening kit, and measuring fluorescence value by using a multifunctional enzyme-labeling instrument, wherein the excitation wavelength during detection is 560nm, and the emission wavelength is 590nm. And finally, calculating the inhibition rate of the drug to COX-2 at each concentration according to the fluorescence value, drawing a dose-effect relationship graph and calculating an IC50 value.
2.3.2 inhibition (%) = (RFU 100% enzyme activity control-RFU sample)/(RFU 100% enzyme activity control-RFU blank) ×100%.
2.4 inhibition of COX-1 by larch resin alcohol, larch resin alcohol 4-O-beta-D-glucopyranoside
2.4.1 Each test sample was provided with 3 multiple wells, and the fluorescence values were measured using a multifunctional microplate reader with reference to the instructions for use of the cyclooxygenase-1 (COX-1) inhibitor screening kit, and the fluorescence values were measured at 535nm excitation wavelength, 587nm emission wavelength, at 5 minutes and 10 minutes, respectively. And finally, calculating the inhibition rate of the drug to COX-1 at each concentration according to the fluorescence value.
2.4.2 inhibition Rate
slope (sample slope) =Δrfu (RFU 2-RFU 1)/Δt (T2-T1)
Inhibition (%) = (slide 100% enzyme activity control-slide sample)/slide 100% enzyme activity control x 100%.
3. Experimental results
According to FIGS. 1 and 2, the larch resin alcohol and the larch resin alcohol 4-O-beta-D-glucopyranoside have different degrees of inhibition on COX-2, wherein the inhibition on COX-2 by the larch resin alcohol is remarkable, and the IC50 value of the larch resin alcohol is 1.41 mu M and the IC50 value of the larch resin alcohol 4-O-beta-D-glucopyranoside is 2.01 mu M through curve fitting. In contrast, celecoxib has an IC50 value of 41.52nM. As can be seen in FIG. 3, none of the other groups had significant inhibition of COX-1, except the aspirin control group.
4. Conclusion(s)
The IC50 of the tested medicine for inhibiting COX-2 is lower than that of the control medicine, but the maximum effect can ensure that the enzyme can reach saturation, and the medicine has no obvious COX-1 inhibition activity, is a Chinese herbal medicine with long medicinal history, has higher safety and has the potential of being developed into COX-2 inhibitor medicines and health care products.
Example 3: effect of larch resin alcohol, larch resin alcohol 4-O-beta-D-glucopyranoside on RAW264.7 cell proliferation
1. Preparation of experimental materials
Experimental cell lines: mouse macrophage RAW264.7, purchased from Shanghai ATCC cell bank. Celecoxib, available from Shanghai Ara Ding Shenghua technologies, inc., lot C20140419; aspirin, lot number L2017111, available from shanghai alaa Ding Shenghua technologies inc; DMEM, PBS, DMSO penicillin and streptomycin (P/S) were purchased from Solarbio; fetal Bovine Serum (FBS) was purchased from Biosharp; trypsin digests were purchased from Gibco; CCK8 was purchased from the syn institute; infinite 200PRO multifunctional full-wavelength microplate reader, tecan, australia; MCO-20AIC carbon dioxide incubator, juferon, USA.
2. Experimental method
Cell culture medium consisted of DMEM, 10% FBS, 1% P/S, RAW264.7 cells were cultured in an incubator at 37℃with 5% CO 2. The medium was changed every 2 days, cells were observed under a mirror to grow to 85% and digested with 0.25% pancreatin and treated to a cell suspension for the experiment. 5X 104 cells/mL of RAW264.7 cells were seeded in 96-well plates with 100. Mu.L of cell suspension per well. After 24h of culture, the cells were used for the subsequent experiments when they adhered well and were confluent at the bottom of 96-well plates. Dissolving larch resin alcohol, larch resin alcohol 4-O-beta-D-glucopyranoside and celecoxib prepared in the method of example 1 by using DMSO, and selecting the larch resin alcohol and the larch resin alcohol 4-O-beta-D-glucopyranoside with final concentrations of 7.8125, 15.625, 31.25, 62.5, 125, 250, 500 and 1000 mu M in a test; 0.0625, 0.125, 0.25, 0.5, 1, 2, 4, 8mM aspirin; 0.0625, 0.125, 0.25, 0.5, 1, 2, 4, 8. Mu.M celecoxib, 6 wells per group, after 24h incubation in an incubator with 5% CO2 at 37℃old medium was discarded, 100. Mu.L of basal DMEM medium containing 10% CCK8 was added, after 2h incubation the absorbance was measured at 450nm in an enzyme-labelling apparatus and the cell viability of different concentrations of larch resin alcohol, larch resin alcohol 4-O-. Beta. -D-glucopyranoside and celecoxib relative to the blank group was calculated.
3. Statistics and data analysis: all experimental data were initially collated with Excel 2010 software and One-way anova using SPSS26.0 software was used for One-way analysis of variance, with results expressed as "mean ± standard deviation".
4. Experimental results
As shown in FIG. 4, the CCK8 results showed that the concentrations of 0-500. Mu.M of larch resin alcohol and larch resin alcohol 4-O-. Beta. -D-glucopyranoside had no significant effect on cell proliferation, and therefore, 0-500. Mu.M of larch resin alcohol, larch resin alcohol 4-O-. Beta. -D-glucopyranoside, 1mM of aspirin and 1. Mu.M of celecoxib were selected for the subsequent experiments. The larch resin alcohol and the 4-O-beta-D-glucopyranoside of the larch resin alcohol have weak cytotoxicity and larger dosage interval.
Example 4: COX-2 selective inhibition of LPS-induced RAW264.7 cell model by lariciresinol, lariciresinol 4-O-beta-D-glucopyranoside
1. Preparation of experimental materials
Experimental cell lines: mouse macrophage RAW264.7, purchased from Shanghai ATCC cell bank. Celecoxib, available from Shanghai Ara Ding Shenghua technologies, inc., lot C20140419; aspirin, lot number L2017111, available from shanghai alaa Ding Shenghua technologies inc; DMEM, PBS, DMSO penicillin and streptomycin (P/S) were purchased from Solarbio; fetal Bovine Serum (FBS) was purchased from Biosharp; trypsin digests were purchased from Gibco; LPS was purchased from Shanghai Ala Di Ltd, and PGE2 detection kit was purchased from Rui Xin Bio; infinite 200PRO multifunctional full-wavelength microplate reader, tecan, australia; MCO-20AIC carbon dioxide incubator, juferon, USA.
2. Experimental method
Cell culture medium consisted of DMEM, 10% FBS, 1% P/S, RAW264.7 cells were cultured in an incubator at 37℃with 5% CO 2. The medium was changed every 2 days, cells were observed under a mirror to grow to 85% and digested with 0.25% pancreatin and treated to a cell suspension for the experiment. Cells were seeded into 24-well plates at a density of 4×105 cells/well. After 24h of culture, the cells were used for the subsequent experiments when they adhered well and were confluent at the bottom of the cell plate. The tests were divided into aspirin groups, celecoxib groups, larch resin alcohol prepared by the method of example 1 with different concentrations, and larch resin alcohol 4-O-beta-D-glucopyranoside groups, each group being repeated 3 times. Wherein, the final concentration of aspirin is 1mM, the final concentration of celecoxib group is 1 mu M, and the larch resin alcohol, the larch resin alcohol 4-O-beta-D-glucopyranoside group with different concentrations are added with the corresponding mass of larch resin alcohol, the larch resin alcohol 4-O-beta-D-glucopyranoside to make the final concentration 500 mu M, 250 mu M, 125 mu M, 62.5 mu M, 31.25 mu M and 15.625 mu M. LPS was added to the cells at a final concentration of 1.0. Mu.g/mL at the time of the experiment, stimulated for 12h in an incubator at 37℃with 5% CO2, after which the drug was given to the different groups for further incubation for 24h.
The cell culture supernatant was collected, and the steps of measuring PGE2 and PGF1α in the supernatant were performed with reference to the instructions of the ELISA kit.
COX-2 inhibition% = (PGE 2 after drug action of control group PGE 2-C)/control group PGE 2X 100%.
COX-1 inhibition% = (PGF1α after drug action of control PGF1α—C)/control PGF1α×100% of control PGF1α.
Statistics and data analysis: all experimental data were initially collated with Excel 2010 software and One-way anova using SPSS26.0 software was used for One-way analysis of variance, with results expressed as "mean ± standard deviation".
3. Experimental results
As shown in FIG. 5, the larch resin alcohol 4-O-beta-D-glucopyranoside and the positive control agent have remarkable inhibition effect on COX-2 in the LPS-induced RAW264.7 cell model, and 100 mu M of larch resin alcohol, the larch resin alcohol 4-O-beta-D-glucopyranoside group and the celecoxib group can achieve the saturation inhibition rate of enzymes, and the inhibition rate of aspirin is lower than that of the larch resin alcohol and the larch resin alcohol 4-O-beta-D-glucopyranoside group. Furthermore, as the COX-2 inhibition rate decreased with decreasing concentrations of larch resin alcohol, larch resin alcohol 4-O-beta-D-glucopyranoside, larch resin alcohol, 4-O-beta-D-glucopyranoside, may dose-dependently inhibit COX-2 activity in the LPS-induced RAW264.7 cell model.
As shown in FIG. 6, the inhibition effect of the aspirin group on COX-1 was remarkable, and the celecoxib group and the larch resin alcohol, the larch resin alcohol 4-O-beta-D-glucopyranoside at different concentrations hardly inhibited COX-1. Larch resin alcohol, larch resin alcohol 4-O-beta-D-glucopyranoside, has selective inhibition on COX-2.
Example 5: preparation of larch resin alcohol freeze-dried powder injection (1000 bottles)
Prescription: 8 g of larch resin alcohol prepared by the method of example 1 and 80 g of tertiary butanol; 80 g of mannitol, 10g of glycine and 2000ml of water for injection.
The preparation method comprises the following steps: precisely weighing larch resin alcohol with a prescription amount, adding tertiary butanol with the prescription amount, and stirring for dissolution to obtain a solution A; adding 10g glycine into the solution A, and stirring and dissolving to obtain a solution B; adding mannitol with a prescription amount into 1500ml of water for injection, and dissolving to obtain a solution C; adding the solution B into the solution C, and adjusting the pH to 6.5 by using sodium bicarbonate and hydrochloric acid; supplementing water for injection to the prescription amount to obtain larch resin alcohol solution; filtering, packaging, and freeze-drying to obtain larch resin alcohol freeze-dried powder injection. The freeze drying method comprises the following steps: pre-freezing: maintaining at 0deg.C for 2 hr, and cooling to-40deg.C for 4 hr; primary sublimation drying: -40 ℃ for 12h, -35 ℃ for 10h, -25 ℃ for 18h, -20 ℃ for 6h, -10 ℃ for 3h, and 0 ℃ for 2h; and (3) secondary drying: the temperature was kept at 25℃for 8 hours. The obtained preparation has stable process and controllable quality, and meets the quality inspection requirement of freeze-dried preparations.
Example 6: preparation of tablet of Fall She Songshu fatty alcohol 4-O-beta-D-glucopyranoside (1000 tablets)
Prescription: 5.0g of L-She Songshu fatty alcohol 4-O-beta-D-glucopyranoside, 145.0g of lactose, 145.0g of pregelatinized starch, 3.0g of sodium carboxymethyl starch, 3.0g of magnesium stearate and 20.0g of hydroxypropyl methylcellulose, which are prepared by the method of example 1.
The preparation method comprises the following steps: (1) prescription weighing: weighing superfine crushed raw material of the She Songshu fatty alcohol 4-O-beta-D-glucopyranoside according to the formula amount, and sieving for standby. (2) adhesive preparation: the measured hydroxypropyl methylcellulose is added into 200ml of 50% ethanol solution, and is dissolved for standby. (3) granulating and drying: mixing She Songshu fatty alcohol 4-O-beta-D-glucopyranoside, lactose, pregelatinized starch and sodium carboxymethyl starch, slowly adding the above binder, making soft material, standing to obtain a powder, sieving with 18-24 mesh sieve, granulating, and drying at 60deg.C for 4 hr. (4) finishing and total mixing: and (3) sieving the dried granules with a 18-24-mesh screen, grading, and uniformly mixing the granules with the prescription amount of magnesium stearate after grading. (5) tabletting: and tabletting by adopting a shallow concave punch with the diameter of 8.0mm, and adjusting a tabletting machine regulator to ensure that the difference of hardness and tablet weight meets the requirements. The obtained tablet has stable process and controllable quality, and meets the quality inspection requirement of the tablet.
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (7)
1. The use of larch resin alcohol and derivatives thereof in the preparation of COX-2 selective inhibitors, characterized in that: the structural formula of the larch resin alcohol is I, one of the larch resin alcohol derivatives is larch She Songshu resin alcohol 4-O-beta-D-glucopyranoside, the structural formula is II,
2. use of larch resin alcohol and derivatives thereof according to claim 1 for the preparation of COX-2 selective inhibitors, characterized in that: the extraction method of larch resin alcohol and derivatives thereof comprises the following steps: weighing radix Isatidis decoction pieces, sequentially reflux-extracting with 8 times of 80% ethanol for 3 times each for 1 hr, filtering, mixing filtrates, recovering ethanol, and extracting with n-butanol to obtain extract; loading into a glass chromatographic column, adding 2 times of distilled water into the extract, suspending in an ultrasonic instrument, filtering, loading into a D101 macroporous adsorption resin column, eluting the sample with 10%, 20%, 30%, 40%, 50% and 60% ethanol sequentially, taking 500mL as a unit, collecting the fraction, concentrating, and drying to obtain a total lignanoid sample of the radix isatidis; separating the sample by octadecyl silane chemically bonded (C18) column chromatography, eluting with 7%, 20%, 30%, 40%, 50%, and pure methanol, detecting the sample by HPLC, and mixing the eluted samples according to HPLC profile data to obtain larch resin alcohol.
3. Use of larch resin alcohol and derivatives thereof according to claim 1 for the preparation of COX-2 selective inhibitors, characterized in that: the extraction method of larch resin alcohol and derivatives thereof comprises the following steps: weighing radix Isatidis decoction pieces, sequentially reflux-extracting with 8 times of 80% ethanol for 3 times each for 1 hr, filtering, mixing filtrates, recovering ethanol, and extracting with n-butanol to obtain extract; loading into a glass chromatographic column, adding 2 times of distilled water into the extract, suspending in an ultrasonic instrument, filtering, loading into a D101 macroporous adsorption resin column, eluting the sample with 10%, 20%, 30%, 40%, 50% and 60% ethanol sequentially, taking 500mL as a unit, collecting the fraction, concentrating, and drying to obtain a total lignanoid sample of the radix isatidis; separating the sample by octadecyl silane chemically bonded (C18) column chromatography, eluting with 7%, 20%, 30%, 40%, 50%, and pure methanol, detecting the sample by HPLC, and mixing the eluted samples according to HPLC profile data to obtain larch resin alcohol 4-O-beta-D-glucopyranoside.
4. Use of larch resin alcohol and derivatives thereof according to claim 1 for the preparation of COX-2 selective inhibitors, characterized in that: the larch resin alcohol derivative is one of polyglycoside, ester and salt containing larch resin alcohol group.
5. Use of larch resin alcohol and derivatives thereof according to claim 1 for the preparation of COX-2 selective inhibitors, characterized in that: the larch resin alcohol and the derivative thereof are added with a pharmaceutically acceptable carrier or excipient to prepare the preparation.
6. Use of larch resin alcohol and derivatives thereof according to claim 5 for the preparation of COX-2 selective inhibitors, wherein: the preparation is one of injection, lyophilized powder for injection, injection microsphere, liposome, bilayer/multilayer tablet, buccal tablet, sublingual tablet, capsule, aqua, powder, paste, spray, granule, soft capsule, dripping pill, gel, patch, and paste.
7. Use of larch resin alcohol and derivatives thereof according to claim 1 for the preparation of COX-2 selective inhibitors, characterized in that: application of COX-2 selective inhibitor in preparing antipyretic, antiinflammatory, analgesic and antitumor drugs is provided.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311637681.8A CN117503785A (en) | 2023-12-01 | 2023-12-01 | Application of larch resin alcohol and derivative thereof in preparation of COX-2 selective inhibitor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311637681.8A CN117503785A (en) | 2023-12-01 | 2023-12-01 | Application of larch resin alcohol and derivative thereof in preparation of COX-2 selective inhibitor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117503785A true CN117503785A (en) | 2024-02-06 |
Family
ID=89758619
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311637681.8A Pending CN117503785A (en) | 2023-12-01 | 2023-12-01 | Application of larch resin alcohol and derivative thereof in preparation of COX-2 selective inhibitor |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117503785A (en) |
-
2023
- 2023-12-01 CN CN202311637681.8A patent/CN117503785A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5410683B2 (en) | Hepatoprotective agent and anti-TNF-α agonist obtained from Kankaniku Juyo | |
| CN102526165A (en) | Rhodiola effective fractions, preparation method, drug composition and uses thereof | |
| CN102397269A (en) | Application of chalcone compounds in preparations of inflammation resisting medicines | |
| WO2011047576A1 (en) | Use of albiflorin for anti-depression | |
| WO2015192758A1 (en) | Anti-tumor pharmaceutical application of pentacyclic triterpene saponin compounds of szechuan melandium root | |
| CN102731276B (en) | Diterpene compound possessing antitumor activity, preparation method thereof and application thereof | |
| TWI782301B (en) | Combination product containing a limonoid and a DPP-4 inhibitor | |
| CN101849950A (en) | Application of rotundic acid in preparing blood lipid regulating medicines | |
| KR100979459B1 (en) | Tetracera scandens extracts and 4H-chromen-4-one derivatives isolated therefrom increasing glucose uptake in differentiated L6 muscle cells | |
| CN108314620B (en) | Sapium sebiferum elements I and J, and pharmaceutical composition and application thereof | |
| KR101706156B1 (en) | A composition comprising compounds isolated from Smilax china for preventing or treating metabolic disorder | |
| EP1222925B1 (en) | Kavalactone as TNF-alpha production inhibitor | |
| US10966996B2 (en) | Glechoma longitube extract, preparation method for same, and use thereof in sugar reduction, weight loss, and lipid reduction | |
| JP7326561B1 (en) | Use of an extract of the active site of Gardenia japonicum in the preparation of a medicament for treating inflammatory diseases or tumors | |
| CN117503785A (en) | Application of larch resin alcohol and derivative thereof in preparation of COX-2 selective inhibitor | |
| CN101129394A (en) | New use of aesculin in preventing and/or treating cardiovascular disease | |
| CN114748518B (en) | Oral preparation containing caffeic acid ester and breviscapine for treating intestinal cancer, and its preparation method | |
| CN113956229B (en) | Lignan compound in lilac and preparation method and application thereof | |
| CN105796764B (en) | Preparation method and application of negundo chastetree fruit total lignans | |
| CN112870208A (en) | Application of Pubescenoside A in preparation of medicine for preventing and treating myocardial ischemia-reperfusion injury | |
| CN114377023A (en) | Preparation of dianthrone compound and application of dianthrone compound in preventing and treating insulin resistance-related metabolic diseases such as diabetes, hyperlipidemia and the like | |
| WO2004009065A1 (en) | Hypoglycemic agent, liver protecting agent and anticancer agent containing lignans originating in hongdoushan | |
| CN109810084B (en) | Paeonilactiflorol, pharmaceutical composition thereof, preparation method and application thereof | |
| CN110279866A (en) | Combination product comprising limonoid and Thiazolidinediones | |
| US10329316B2 (en) | Phenylpropanoid compound and preparation method and use thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |