CN102924554B - C-3,11,12,20-tetrasubstituted C-21 steroid derivative and its pharmaceutical composition and use in medicine - Google Patents
C-3,11,12,20-tetrasubstituted C-21 steroid derivative and its pharmaceutical composition and use in medicine Download PDFInfo
- Publication number
- CN102924554B CN102924554B CN201210419579.6A CN201210419579A CN102924554B CN 102924554 B CN102924554 B CN 102924554B CN 201210419579 A CN201210419579 A CN 201210419579A CN 102924554 B CN102924554 B CN 102924554B
- Authority
- CN
- China
- Prior art keywords
- group
- cym
- zebra fish
- acid
- pyrans glycosyl
- 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.)
- Active
Links
- 239000003814 drug Substances 0.000 title claims abstract description 24
- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 8
- -1 C-21 steroid Chemical class 0.000 title abstract description 32
- 229930182470 glycoside Natural products 0.000 claims abstract description 57
- 150000002338 glycosides Chemical class 0.000 claims abstract description 56
- 150000003839 salts Chemical class 0.000 claims abstract description 18
- 238000002360 preparation method Methods 0.000 claims abstract description 15
- 230000003556 anti-epileptic effect Effects 0.000 claims description 48
- 239000001961 anticonvulsive agent Substances 0.000 claims description 30
- 230000001037 epileptic effect Effects 0.000 claims description 9
- 239000003937 drug carrier Substances 0.000 claims description 2
- 150000003431 steroids Chemical class 0.000 claims 3
- 206010015037 epilepsy Diseases 0.000 abstract description 35
- 125000002887 hydroxy group Chemical group [H]O* 0.000 abstract description 34
- 229910052739 hydrogen Inorganic materials 0.000 abstract description 23
- 239000001257 hydrogen Substances 0.000 abstract description 23
- 125000004185 ester group Chemical group 0.000 abstract description 9
- 125000002252 acyl group Chemical group 0.000 abstract description 6
- 125000003147 glycosyl group Chemical group 0.000 abstract description 2
- 125000004442 acylamino group Chemical group 0.000 abstract 3
- 125000003277 amino group Chemical group 0.000 abstract 3
- 239000004480 active ingredient Substances 0.000 abstract 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 abstract 1
- 241000252212 Danio rerio Species 0.000 description 132
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 99
- 150000004880 oxines Chemical class 0.000 description 76
- 230000033001 locomotion Effects 0.000 description 60
- 150000001875 compounds Chemical class 0.000 description 49
- 210000001161 mammalian embryo Anatomy 0.000 description 49
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 45
- 238000000034 method Methods 0.000 description 36
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 34
- 241000587240 Cynanchum Species 0.000 description 34
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 32
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 27
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 27
- 230000001225 therapeutic effect Effects 0.000 description 27
- 230000008569 process Effects 0.000 description 26
- 241000893871 Dregea Species 0.000 description 25
- 206010010904 Convulsion Diseases 0.000 description 24
- CWRVKFFCRWGWCS-UHFFFAOYSA-N Pentrazole Chemical compound C1CCCCC2=NN=NN21 CWRVKFFCRWGWCS-UHFFFAOYSA-N 0.000 description 24
- 206010002091 Anaesthesia Diseases 0.000 description 21
- 230000037005 anaesthesia Effects 0.000 description 21
- 238000000605 extraction Methods 0.000 description 21
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 18
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 17
- RYVMUASDIZQXAA-UHFFFAOYSA-N pyranoside Natural products O1C2(OCC(C)C(OC3C(C(O)C(O)C(CO)O3)O)C2)C(C)C(C2(CCC3C4(C)CC5O)C)C1CC2C3CC=C4CC5OC(C(C1O)O)OC(CO)C1OC(C1OC2C(C(OC3C(C(O)C(O)C(CO)O3)O)C(O)C(CO)O2)O)OC(CO)C(O)C1OC1OCC(O)C(O)C1O RYVMUASDIZQXAA-UHFFFAOYSA-N 0.000 description 17
- DKVBOUDTNWVDEP-NJCHZNEYSA-N teicoplanin aglycone Chemical compound N([C@H](C(N[C@@H](C1=CC(O)=CC(O)=C1C=1C(O)=CC=C2C=1)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)OC=1C=C3C=C(C=1O)OC1=CC=C(C=C1Cl)C[C@H](C(=O)N1)NC([C@H](N)C=4C=C(O5)C(O)=CC=4)=O)C(=O)[C@@H]2NC(=O)[C@@H]3NC(=O)[C@@H]1C1=CC5=CC(O)=C1 DKVBOUDTNWVDEP-NJCHZNEYSA-N 0.000 description 17
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 16
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 16
- 239000002253 acid Substances 0.000 description 16
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 16
- 229960002790 phenytoin sodium Drugs 0.000 description 16
- FJPYVLNWWICYDW-UHFFFAOYSA-M sodium;5,5-diphenylimidazolidin-1-ide-2,4-dione Chemical compound [Na+].O=C1[N-]C(=O)NC1(C=1C=CC=CC=1)C1=CC=CC=C1 FJPYVLNWWICYDW-UHFFFAOYSA-M 0.000 description 16
- 238000011278 co-treatment Methods 0.000 description 15
- 239000007788 liquid Substances 0.000 description 15
- 239000000243 solution Substances 0.000 description 15
- 241000251468 Actinopterygii Species 0.000 description 14
- 239000003480 eluent Substances 0.000 description 14
- 239000000284 extract Substances 0.000 description 14
- 230000004720 fertilization Effects 0.000 description 14
- 238000010898 silica gel chromatography Methods 0.000 description 14
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 13
- 101100115553 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) cym-1 gene Proteins 0.000 description 12
- 238000004458 analytical method Methods 0.000 description 12
- 239000013641 positive control Substances 0.000 description 12
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 11
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 11
- 150000002500 ions Chemical class 0.000 description 11
- 229960004002 levetiracetam Drugs 0.000 description 11
- HPHUVLMMVZITSG-ZCFIWIBFSA-N levetiracetam Chemical compound CC[C@H](C(N)=O)N1CCCC1=O HPHUVLMMVZITSG-ZCFIWIBFSA-N 0.000 description 11
- 239000000203 mixture Substances 0.000 description 11
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 10
- 238000004445 quantitative analysis Methods 0.000 description 10
- 239000000741 silica gel Substances 0.000 description 10
- 229910002027 silica gel Inorganic materials 0.000 description 10
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 9
- 230000003542 behavioural effect Effects 0.000 description 9
- 235000015165 citric acid Nutrition 0.000 description 9
- 150000002431 hydrogen Chemical class 0.000 description 9
- 229910052500 inorganic mineral Inorganic materials 0.000 description 9
- 239000011707 mineral Substances 0.000 description 9
- 150000007524 organic acids Chemical class 0.000 description 9
- 235000006408 oxalic acid Nutrition 0.000 description 9
- 239000002904 solvent Substances 0.000 description 9
- 229940095064 tartrate Drugs 0.000 description 9
- 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 8
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 8
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 8
- 229960003965 antiepileptics Drugs 0.000 description 8
- 229960000648 digitoxin Drugs 0.000 description 8
- 235000019253 formic acid Nutrition 0.000 description 8
- 125000000468 ketone group Chemical group 0.000 description 8
- 239000000463 material Substances 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical group CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 7
- 241001465754 Metazoa Species 0.000 description 7
- 238000009395 breeding Methods 0.000 description 7
- 230000001488 breeding effect Effects 0.000 description 7
- 239000002026 chloroform extract Substances 0.000 description 7
- 230000002508 compound effect Effects 0.000 description 7
- 210000002257 embryonic structure Anatomy 0.000 description 7
- 238000011156 evaluation Methods 0.000 description 7
- 238000009472 formulation Methods 0.000 description 7
- JEGUKCSWCFPDGT-UHFFFAOYSA-N h2o hydrate Chemical compound O.O JEGUKCSWCFPDGT-UHFFFAOYSA-N 0.000 description 7
- 230000013011 mating Effects 0.000 description 7
- 239000013642 negative control Substances 0.000 description 7
- 230000000050 nutritive effect Effects 0.000 description 7
- 239000000843 powder Substances 0.000 description 7
- 238000000746 purification Methods 0.000 description 7
- 238000001223 reverse osmosis Methods 0.000 description 7
- 229940072040 tricaine Drugs 0.000 description 7
- FQZJYWMRQDKBQN-UHFFFAOYSA-N tricaine methanesulfonate Chemical compound CS([O-])(=O)=O.CCOC(=O)C1=CC=CC([NH3+])=C1 FQZJYWMRQDKBQN-UHFFFAOYSA-N 0.000 description 7
- 210000001325 yolk sac Anatomy 0.000 description 7
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 6
- 238000005160 1H NMR spectroscopy Methods 0.000 description 6
- 238000004440 column chromatography Methods 0.000 description 6
- 239000003921 oil Substances 0.000 description 6
- 239000003981 vehicle Substances 0.000 description 6
- 241000345998 Calamus manan Species 0.000 description 5
- 238000004992 fast atom bombardment mass spectroscopy Methods 0.000 description 5
- 239000000499 gel Substances 0.000 description 5
- 150000008131 glucosides Chemical group 0.000 description 5
- 239000003208 petroleum Substances 0.000 description 5
- 239000000546 pharmaceutical excipient Substances 0.000 description 5
- 235000012950 rattan cane Nutrition 0.000 description 5
- 230000002441 reversible effect Effects 0.000 description 5
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Natural products OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 4
- ODFJOVXVLFUVNQ-UHFFFAOYSA-N acetarsol Chemical compound CC(=O)NC1=CC([As](O)(O)=O)=CC=C1O ODFJOVXVLFUVNQ-UHFFFAOYSA-N 0.000 description 4
- 239000002775 capsule Substances 0.000 description 4
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 229930194991 otophylloside Natural products 0.000 description 4
- APHQTSDXMVIBRQ-UHFFFAOYSA-N otophylloside H Natural products COC1CC(OC2C(O)CC(OC3CCC4(C)C5CC(OC(=O)c6ccc(O)cc6)C7(C)C(O)(CCC7(O)C5(O)CC=C4C3)C(=O)C)OC2C)OC(C)C1OC8CC(OC)C(OC9OC(CO)C(OC%10OC(CO)C(O)C(O)C%10O)C(O)C9O)C(C)O8 APHQTSDXMVIBRQ-UHFFFAOYSA-N 0.000 description 4
- 229960003742 phenol Drugs 0.000 description 4
- 208000024891 symptom Diseases 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 230000001419 dependent effect Effects 0.000 description 3
- HAPOVYFOVVWLRS-UHFFFAOYSA-N ethosuximide Chemical compound CCC1(C)CC(=O)NC1=O HAPOVYFOVVWLRS-UHFFFAOYSA-N 0.000 description 3
- 229930182478 glucoside Natural products 0.000 description 3
- 239000008187 granular material Substances 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000007689 inspection Methods 0.000 description 3
- 230000007774 longterm Effects 0.000 description 3
- DDBREPKUVSBGFI-UHFFFAOYSA-N phenobarbital Chemical compound C=1C=CC=CC=1C1(CC)C(=O)NC(=O)NC1=O DDBREPKUVSBGFI-UHFFFAOYSA-N 0.000 description 3
- DQMZLTXERSFNPB-UHFFFAOYSA-N primidone Chemical compound C=1C=CC=CC=1C1(CC)C(=O)NCNC1=O DQMZLTXERSFNPB-UHFFFAOYSA-N 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- AEQFSUDEHCCHBT-UHFFFAOYSA-M sodium valproate Chemical compound [Na+].CCCC(C([O-])=O)CCC AEQFSUDEHCCHBT-UHFFFAOYSA-M 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 238000007619 statistical method Methods 0.000 description 3
- 208000005809 status epilepticus Diseases 0.000 description 3
- 230000003637 steroidlike Effects 0.000 description 3
- 238000002560 therapeutic procedure Methods 0.000 description 3
- XMIIGOLPHOKFCH-UHFFFAOYSA-N 3-phenylpropionic acid Chemical compound OC(=O)CCC1=CC=CC=C1 XMIIGOLPHOKFCH-UHFFFAOYSA-N 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 2
- 101100068867 Caenorhabditis elegans glc-1 gene Proteins 0.000 description 2
- 101100068866 Caenorhabditis elegans glc-2 gene Proteins 0.000 description 2
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 2
- CPELXLSAUQHCOX-UHFFFAOYSA-N Hydrogen bromide Chemical compound Br CPELXLSAUQHCOX-UHFFFAOYSA-N 0.000 description 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
- CXOFVDLJLONNDW-UHFFFAOYSA-N Phenytoin Chemical compound N1C(=O)NC(=O)C1(C=1C=CC=CC=1)C1=CC=CC=C1 CXOFVDLJLONNDW-UHFFFAOYSA-N 0.000 description 2
- 208000005392 Spasm Diseases 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- KJADKKWYZYXHBB-XBWDGYHZSA-N Topiramic acid Chemical compound C1O[C@@]2(COS(N)(=O)=O)OC(C)(C)O[C@H]2[C@@H]2OC(C)(C)O[C@@H]21 KJADKKWYZYXHBB-XBWDGYHZSA-N 0.000 description 2
- 208000003443 Unconsciousness Diseases 0.000 description 2
- 208000016807 X-linked intellectual disability-macrocephaly-macroorchidism syndrome Diseases 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 239000000443 aerosol Substances 0.000 description 2
- 125000003368 amide group Chemical group 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- NIJJYAXOARWZEE-UHFFFAOYSA-N di-n-propyl-acetic acid Natural products CCCC(C(O)=O)CCC NIJJYAXOARWZEE-UHFFFAOYSA-N 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- ZEIXWOCIFAPPPJ-UHFFFAOYSA-N dregeoside Natural products COC1CC(OC2C(C)OC(CC2OC)OC3C(C)OC(CC3OC)OC4CCC5(C)C(CCC6(O)C5CC(OC(=O)c7ccccc7)C8(C)C(O)(CCC68O)C(C)O)C4)OC(C)C1O ZEIXWOCIFAPPPJ-UHFFFAOYSA-N 0.000 description 2
- 229960002767 ethosuximide Drugs 0.000 description 2
- 230000004438 eyesight Effects 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- MNWFXJYAOYHMED-UHFFFAOYSA-N heptanoic acid Chemical compound CCCCCCC(O)=O MNWFXJYAOYHMED-UHFFFAOYSA-N 0.000 description 2
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 229960001848 lamotrigine Drugs 0.000 description 2
- PYZRQGJRPPTADH-UHFFFAOYSA-N lamotrigine Chemical compound NC1=NC(N)=NN=C1C1=CC=CC(Cl)=C1Cl PYZRQGJRPPTADH-UHFFFAOYSA-N 0.000 description 2
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 2
- CTRLABGOLIVAIY-UHFFFAOYSA-N oxcarbazepine Chemical compound C1C(=O)C2=CC=CC=C2N(C(=O)N)C2=CC=CC=C21 CTRLABGOLIVAIY-UHFFFAOYSA-N 0.000 description 2
- 230000000803 paradoxical effect Effects 0.000 description 2
- 235000019260 propionic acid Nutrition 0.000 description 2
- 238000004064 recycling Methods 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 229940084026 sodium valproate Drugs 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000000829 suppository Substances 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- 229960004394 topiramate Drugs 0.000 description 2
- RSJKGSCJYJTIGS-UHFFFAOYSA-N undecane Chemical compound CCCCCCCCCCC RSJKGSCJYJTIGS-UHFFFAOYSA-N 0.000 description 2
- LSPHULWDVZXLIL-LDWIPMOCSA-N (?)-Camphoric acid Chemical compound CC1(C)[C@@H](C(O)=O)CC[C@@]1(C)C(O)=O LSPHULWDVZXLIL-LDWIPMOCSA-N 0.000 description 1
- MIOPJNTWMNEORI-GMSGAONNSA-N (S)-camphorsulfonic acid Chemical compound C1C[C@@]2(CS(O)(=O)=O)C(=O)C[C@@H]1C2(C)C MIOPJNTWMNEORI-GMSGAONNSA-N 0.000 description 1
- FEJIIZAOQRTGPC-UHFFFAOYSA-N 5,5-diphenylimidazolidin-4-one Chemical compound O=C1NCNC1(C=1C=CC=CC=1)C1=CC=CC=C1 FEJIIZAOQRTGPC-UHFFFAOYSA-N 0.000 description 1
- JWMFYGXQPXQEEM-NUNROCCHSA-N 5β-pregnane Chemical compound C([C@H]1CC2)CCC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H](CC)[C@@]2(C)CC1 JWMFYGXQPXQEEM-NUNROCCHSA-N 0.000 description 1
- 206010061623 Adverse drug reaction Diseases 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 241000208327 Apocynaceae Species 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 1
- 208000018152 Cerebral disease Diseases 0.000 description 1
- 208000017667 Chronic Disease Diseases 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
- ITPJPPQJMRCTTO-UHFFFAOYSA-N Dregeoside Da1 Natural products COC1CC(OC2CCC3(C)C4C(O)C(O)C5(C)C(CCC5(O)C4CC=C3C2)C(C)O)OC(C)C1OC1CC(OC)C(OC2OC(C)C(O)C(OC)C2O)C(C)O1 ITPJPPQJMRCTTO-UHFFFAOYSA-N 0.000 description 1
- 208000030453 Drug-Related Side Effects and Adverse reaction Diseases 0.000 description 1
- 208000012661 Dyskinesia Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 208000002877 Epileptic Syndromes Diseases 0.000 description 1
- 108010010803 Gelatin Proteins 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
- XYZZKVRWGOWVGO-UHFFFAOYSA-N Glycerol-phosphate Chemical class OP(O)(O)=O.OCC(O)CO XYZZKVRWGOWVGO-UHFFFAOYSA-N 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 101000584505 Homo sapiens Synaptic vesicle glycoprotein 2A Proteins 0.000 description 1
- 206010020751 Hypersensitivity 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
- 108010076876 Keratins Proteins 0.000 description 1
- 102000011782 Keratins Human genes 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- 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 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 241000978776 Senegalia senegal Species 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 102100030701 Synaptic vesicle glycoprotein 2A Human genes 0.000 description 1
- 102000004874 Synaptophysin Human genes 0.000 description 1
- 108090001076 Synaptophysin Proteins 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 235000011054 acetic acid Nutrition 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000002082 anti-convulsion Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- BLCFCVYGFDLOSC-UHFFFAOYSA-N benzene;benzenesulfonic acid Chemical compound C1=CC=CC=C1.OS(=O)(=O)C1=CC=CC=C1 BLCFCVYGFDLOSC-UHFFFAOYSA-N 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
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 229960000623 carbamazepine Drugs 0.000 description 1
- FFGPTBGBLSHEPO-UHFFFAOYSA-N carbamazepine Chemical compound C1=CC2=CC=CC=C2N(C(=O)N)C2=CC=CC=C21 FFGPTBGBLSHEPO-UHFFFAOYSA-N 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000002490 cerebral effect Effects 0.000 description 1
- SIHHLZPXQLFPMC-UHFFFAOYSA-N chloroform;methanol;hydrate Chemical compound O.OC.ClC(Cl)Cl SIHHLZPXQLFPMC-UHFFFAOYSA-N 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000008119 colloidal silica Substances 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 229940089052 depakene Drugs 0.000 description 1
- 229940064790 dilantin Drugs 0.000 description 1
- GXGAKHNRMVGRPK-UHFFFAOYSA-N dimagnesium;dioxido-bis[[oxido(oxo)silyl]oxy]silane Chemical compound [Mg+2].[Mg+2].[O-][Si](=O)O[Si]([O-])([O-])O[Si]([O-])=O GXGAKHNRMVGRPK-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- MOTZDAYCYVMXPC-UHFFFAOYSA-N dodecyl hydrogen sulfate Chemical compound CCCCCCCCCCCCOS(O)(=O)=O MOTZDAYCYVMXPC-UHFFFAOYSA-N 0.000 description 1
- 229950004453 doxenitoin Drugs 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- CCIVGXIOQKPBKL-UHFFFAOYSA-N ethanesulfonic acid Chemical compound CCS(O)(=O)=O CCIVGXIOQKPBKL-UHFFFAOYSA-N 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229910000042 hydrogen bromide Inorganic materials 0.000 description 1
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 description 1
- 229940071870 hydroiodic acid Drugs 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 239000003978 infusion fluid Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 239000000391 magnesium silicate Substances 0.000 description 1
- 235000019793 magnesium trisilicate Nutrition 0.000 description 1
- 229940099273 magnesium trisilicate Drugs 0.000 description 1
- 229910000386 magnesium trisilicate Inorganic materials 0.000 description 1
- 206010025482 malaise Diseases 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000007510 mood change Effects 0.000 description 1
- 230000017311 musculoskeletal movement, spinal reflex action Effects 0.000 description 1
- 229940090010 mysoline Drugs 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 229960001816 oxcarbazepine Drugs 0.000 description 1
- 230000001314 paroxysmal effect Effects 0.000 description 1
- 229920003175 pectinic acid Polymers 0.000 description 1
- 238000005453 pelletization Methods 0.000 description 1
- RGSFGYAAUTVSQA-UHFFFAOYSA-N pentamethylene Natural products C1CCCC1 RGSFGYAAUTVSQA-UHFFFAOYSA-N 0.000 description 1
- 229960005152 pentetrazol Drugs 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- 229960002695 phenobarbital Drugs 0.000 description 1
- 229960002036 phenytoin Drugs 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- OXNIZHLAWKMVMX-UHFFFAOYSA-N picric acid Chemical compound OC1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O OXNIZHLAWKMVMX-UHFFFAOYSA-N 0.000 description 1
- IUGYQRQAERSCNH-UHFFFAOYSA-N pivalic acid Chemical compound CC(C)(C)C(O)=O IUGYQRQAERSCNH-UHFFFAOYSA-N 0.000 description 1
- WSHYKIAQCMIPTB-UHFFFAOYSA-M potassium;2-oxo-3-(3-oxo-1-phenylbutyl)chromen-4-olate Chemical compound [K+].[O-]C=1C2=CC=CC=C2OC(=O)C=1C(CC(=O)C)C1=CC=CC=C1 WSHYKIAQCMIPTB-UHFFFAOYSA-M 0.000 description 1
- 230000003518 presynaptic effect Effects 0.000 description 1
- 229960002393 primidone Drugs 0.000 description 1
- JUJWROOIHBZHMG-RALIUCGRSA-N pyridine-d5 Chemical compound [2H]C1=NC([2H])=C([2H])C([2H])=C1[2H] JUJWROOIHBZHMG-RALIUCGRSA-N 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 230000000306 recurrent effect Effects 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000035807 sensation Effects 0.000 description 1
- 235000019615 sensations Nutrition 0.000 description 1
- 230000008786 sensory perception of smell Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 239000008227 sterile water for injection Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 235000012222 talc Nutrition 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 231100001274 therapeutic index Toxicity 0.000 description 1
- 230000008719 thickening Effects 0.000 description 1
- ZMZDMBWJUHKJPS-UHFFFAOYSA-N thiocyanic acid Chemical compound SC#N ZMZDMBWJUHKJPS-UHFFFAOYSA-N 0.000 description 1
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 1
- 229940061414 trileptal Drugs 0.000 description 1
- 239000006216 vaginal suppository Substances 0.000 description 1
- 229940120293 vaginal suppository Drugs 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 229940063682 zarontin Drugs 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides a C-3,11,12,20-tetrasubstituted C-21 steroid derivative shown in the general formula I and its pharmaceutical salt and glycoside, and also provides a pharmaceutical composition containing the C-3,11,12,20-tetrasubstituted C-21 steroid derivative as an active ingredient and a use of the C-3,11,12,20-tetrasubstituted C-21 steroid derivative in preparation of a medicine for treating epilepsy. In the general formula I, R1, R2 and R3 respectively represent a ketonic group, a hydroxyl group, a substituted acyl ester group, an amino-group or a substituted acylamino group; R4 represents a ketonic group, a hydroxyl group, a substituted acyl ester group, an amino-group, a substituted acylamino group, or a glycosyl group; and R5, R6 and R7 respectively represent hydrogen, a hydroxyl group, a substituted acyl ester group, an amino-group or a substituted acylamino group.
Description
Technical field:
The invention belongs to field of medicaments, be specifically related to a kind of natural compounds be activeconstituents antiepileptic drug, more specifically, relate to natural physiological active substance C-3,11,12,17-tetra-replaces C-21 steroid derivative and pharmaceutical composition and its thereof application in the medicine of preparation treatment epileptics.
Background technology:
Epilepsy (epilepsy) is commonly called as " insane crazy, the epilepsy of sheep ", is the electric discharge of cerebral neuron paroxysmal abnormality, causes a kind of chronic disease of of short duration cerebral disorder.This disease due to the neurone position difference in the brain of paradoxical discharge, and has diversified symptom.These symptoms comprise relatively common, significant motor symptoms and the visible loss of consciousness in tetanic old contraction outbreak, old the change observed in multi-motion function in addition, the change of such as sensation, sense of smell, vision and higher function, the such as change of mood, memory, language and vision.Wherein, spasm is one of typical epilepsy syndromes, and the normal involuntary movement with stiff property, indirect, also has not with the seizure types of spasm.In addition, may occur unexpected loss of consciousness, memory, faint suddenly, tic of the limbs, spits out white foams, and the symptom such as to shout, epileptic seizures great majority belong to temporary, and generally namely several points recovered as ordinary person to several tens minutes.But it is long that epilepsy has case history, the feature of recurrent exerbation, the sequela caused thus even can cause disturbance of intelligence.Once suffer from epilepsy, be just difficult to radical cure.Slighter can control outbreak with medicine, and anti-frightened epilepsy agent treatment will be lifelong.When medicine can not control epileptic attack, must operate treatment, but expense, risk are high, and still need to take medicine all the life.
According to WHO statistics, current global epileptic 5,000 ten thousand people, wherein 80% in developing country, annual newly-increased 2,000,000 epileptics.The sickness rate of China's epileptic condition is about 7 ‰, wherein 40% has not accepted any treatment, and what the patient of 40% accepted is non-regular treatment, and annual newly-increased patient 400,000 people nearly.
The antiepileptic drug of WHO recommendation in 1980 mainly contains: phenylethyl barbituric acid (Phenobarbital, Luminal), Carbamzepine (Carbamazepine), phenytoin Sodium (Dilantin, Phenytoin, insanely to stop), Sodium Valproate (Depakene), ethosuximide (Ethosuximide, Zarontin), doxenitoin sodium (Primidonum, Mysoline, Hexamidinu, Primidone).At present, market at home, traditional old kind phenylethyl barbituric acid, Carbamzepine, Sodium Valproate, phenytoin Sodium still occupy the absolute market share.In addition, some antiepileptic Chinese patent medicine products are also had, such as Cynanchum otophvllum sheet, the clear sheet of epilepsy, the peaceful sheet of epilepsy, more epilepsy spirit market sale at home.But these traditional Western medicine side effects are large, and long-term taking can cause larger impact to health.Such as, the First Line medication Carbamzepine for the treatment of epilepsy, this medicine can produce the fragrant Johnson Syndrome (Steven-Johnson syndrome) of severe allergy side effect-Shi Supreme Being, and mortality ratio reaches 1 to 4 and becomes.And Chinese patent medicine product exists the features such as curative effect is remarkable, target spot is unclear, there is no the market competitiveness.
In recent years, in world market, there is multiple new antiepileptic medicine, such as: oxcarbazepine (Trileptal), Levetiracetam (Levetiracetam), topiramate (Topiramate), lamotrigine (Lamotrigine) etc.Wherein, Levetiracetam was in 2008, and sales volume just more than 18.64 hundred million dollars, becomes the cookle in antiepileptic drug.The mechanism of action of this medicine is that synaptophysin SV2A is combined in presynaptic nerve teminal, suppresses the paradoxical discharge in epilepsy loop, thus blocks epileptic seizures.Have that anti-outbreak spectrum is wide, good effect, therapeutic index increase, can with the feature such as other AEDs share.Behind Discussion on Chinese Listed, occupy the market share of Chinese antiepileptic drug 10.65% in 2009.
Therefore, study to send out with external antiepileptic drug and enliven, market growth is rapidly compared, the domestic AED operation listing not having novelty.AED market domestic is at present occupied by import medicine, on the high side, and general patient is difficult to bear, the economic problems that long-term taking brings, and is also difficult to solve China epileptics and seeks medical advice the low problem of rate.Meanwhile, existing drug side effect is all comparatively large, and long-term taking affects greatly.
Up to now, there are no C-3 in prior art, 11,12,20-tetra-replaces the report of C-21 steroid derivatives and pharmacologically active thereof, also has no the report that it has antiepileptic activity.
Summary of the invention:
Object of the present invention aims to provide C-3, and 11,12,20-tetra-replaces C-21 steroid derivative, the pharmaceutical composition being activeconstituents with it, and they treat the application in the medicine of epileptics in preparation.
In order to realize above-mentioned purpose of the present invention, the invention provides following technical scheme:
C-3 shown in general formula (I), 11,12,20-tetra-replace C-21 steroid derivatives and pharmacy acceptable salt thereof and glycoside,
Wherein, R
1, R
2, R
3be independently ketone group, hydroxyl and replacement acyl ester group, amino and replacement amido separately; R
4for ketone group, hydroxyl and replacement acyl ester group, amino and replacement acyl ammonia, glycosyl; R
5, R
6, R
7be independently hydrogen, hydroxyl and replacement acyl ester group thereof, amino and replacement amido separately.
For above-mentioned general formula (I) compound, the preferred technical scheme of the present invention is R
1, R
2be respectively isovaleryl ester group, R
3for acetyl ester group, R
4, R
6for hydroxyl, R
5, R
7be respectively hydrogen.
The present invention further preferred technical scheme is R
1for ketone group, R
2para hydroxybenzene formyl ester group, R
3for hydrogen, R
4for 3-O-β-D-glucopyanosyl base-(1 → 4)-β-D-glucopyanosyl base-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-Folium seu Cortex Nerii pyrans glycosyl-(1 → 4)-β-D-digitoxin pyrans glycosyl, R
5, R
6, R
7be respectively hydroxyl.
The present invention further preferred technical scheme is R
1, R
2, R
3be respectively hydroxyl, R
4for O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
7be respectively hydrogen, R
6for hydroxyl.
The present invention further preferred technical scheme is R
1for ketone group, R
2, R
3be respectively acetyl ester group, R
4for O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
7be respectively hydrogen, R
6for hydroxyl.
The present invention further preferred technical scheme is R
1for ketone group, R
2para hydroxybenzene formyl ester group, R
3for hydrogen, R
4for 3-O-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-Folium seu Cortex Nerii pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-digitoxin pyrans glycosyl, R
5, R
6, R
7be respectively hydroxyl.
The present invention further preferred technical scheme is R
1for cis isopentene ester group, R
2for isovaleryl ester group, R
3for acetyl ester group, R
4for O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
6be respectively hydroxyl, R
7for hydrogen.
The present invention further preferred technical scheme is R
1, R
2, R
3be respectively hydroxyl, R
4for O-β-D-glucopyanosyl base-(1 → 4)-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
6be respectively hydroxyl, R
7for hydrogen.
The present invention further preferred technical scheme is R
1, R
2, R
3be respectively hydroxyl, R
4for O-β-D-glucopyanosyl base-(1 → 4)-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
6be respectively hydroxyl, R
7for hydrogen.
The present invention further preferred technical scheme is R
1for trans isopentene ester group, R
2for isovaleryl ester group, R
3for acetyl ester group, R
4for O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
6be respectively hydroxyl, R
7for hydrogen.
The present invention further preferred technical scheme is R
1, R
3be respectively hydroxyl, R
2for trans isopentene ester group, R
4for O-β-D-glucopyanosyl base-(1 → 4)-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
6be respectively hydroxyl, R
7for hydrogen.
The present invention further preferred technical scheme is R
1, R
3be respectively hydroxyl, R
2for cis isopentene ester group, R
4for O-β-D-glucopyanosyl base-(1 → 4)-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
6be respectively hydroxyl, R
7for hydrogen.
The present invention further preferred technical scheme is R
1, R
2be respectively isovaleryl amino, R
3for kharophen, R
4, R
6for hydroxyl, R
5, R
7be respectively hydrogen.
The present invention further preferred technical scheme is R
1for ketone group, R
2for para hydroxybenzene formamido group, R
3for hydrogen, R
4for O-β-D-Folium seu Cortex Nerii pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
6, R
7be respectively hydroxyl.
The present invention further preferred technical scheme is R
1, R
2, R
3be respectively amino, R
4for O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
7be respectively hydrogen, R
6for hydroxyl.
The present invention further preferred technical scheme is R
1for cis senecioyl is amino, R
2for isovaleryl is amino, R
3for kharophen, R
4for O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
6be respectively hydroxyl, R
7for hydrogen.
The present invention further preferred technical scheme is R
1for trans senecioyl is amino, R
2for isovaleryl is amino, R
3for kharophen, R
4for O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
6be respectively hydroxyl, R
7for hydrogen.
The present invention further preferred technical scheme is R
1, R
3be respectively hydroxyl, R
2for trans senecioyl is amino, R
4for O-β-D-glucopyanosyl base-(1 → 4)-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
6be respectively hydroxyl, R
7for hydrogen.
The present invention further preferred technical scheme is R
1, R
3be respectively amino, R
2for cis senecioyl is amino, R
4for O-β-D-glucopyanosyl base-(1 → 4)-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
6be respectively hydroxyl, R
7for hydrogen.
The present invention further preferred technical scheme is R
1, R
2, R
3be respectively amino, R
4for O-β-D-glucopyanosyl base-(1 → 4)-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
6be respectively hydroxyl, R
7for hydrogen.
The present invention further preferred technical scheme is R
1for ketone group, R
2for para hydroxybenzene formamido group, R
3for hydrogen, R
4for O-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-Folium seu Cortex Nerii pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-digitoxin pyrans glycosyl, R
5, R
6, R
7be respectively hydroxyl.
The present invention further preferred technical scheme is R
1for ketone group, R
2, R
3be respectively kharophen, R
4for O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl, R
5, R
7be respectively hydrogen, R
6for hydroxyl.
Pharmaceutical salts described in above-mentioned any one, refer to pharmacy acceptable salt, comprise the salt formed with organic acid or mineral acid, described organic acid includes but not limited to tartrate, citric acid, formic acid, acetic acid, oxalic acid, butyric acid, oxalic acid, toxilic acid, succsinic acid, hexanodioic acid, alginic acid, citric acid, aspartic acid, benzene Phenylsulfonic acid, dextrocamphoric acid, camphorsulfonic acid, didextrose acid, pentamethylene propionic acid, dodecyl sulphate, ethyl sulfonic acid, glucoheptonic acid, Phosphoric acid glycerol esters, hemisulfic acid, enanthic acid, caproic acid, fumaric acid, 2-ethylenehydrinsulfonic acid, lactic acid, toxilic acid, methylsulfonic acid, nicotinic acid, 2-naphthene sulfonic acid, flutter acid, pectinic acid, 3-phenylpropionic acid, picric acid, PIVALIC ACID CRUDE (25), propionic acid, succsinic acid, tartrate, sulfocyanic acid, p-tosylate and undecane hydrochlorate, described mineral acid includes but not limited to hydrochloric acid, Hydrogen bromide, hydroiodic acid HI, sulfuric acid or phosphoric acid.
The present invention provides the pharmaceutical composition of antiepileptic simultaneously, and it comprises the C-3 according to above-mentioned any one, and 11,12,17-tetra-replaces C-21 steroid derivative or its drug salts or glycoside and the pharmaceutically acceptable carrier of at least one.
The present invention still further provides the C-3 described in above-mentioned any one, and 11,12,17-tetra-replaces the application of C-21 steroid derivative in the medicine of preparation treatment epileptics.
Technique scheme of the present invention proposes based on following discovery and principle.
The compound of C-21 steroidal to be a class with pregnane be parent nucleus, its C-11, C-12, C-20 position hydroxyl often forms ester bond, and C-3 position hydroxyl forms glycoside with the sugar chain formed primarily of α-desoxy sugar.This compounds has neural physiologically active, as anticonvulsion, and anti-epileptic, anti-melancholy etc.The neurosteroid that their chemical structure and physiological function and eighties of last century the nineties are risen is closely similar, and infer thus, C-21 steroidal is a kind of class neurosteroid of plant origin.The present invention is based on the basis of document in early stage, a series of new 11-O-ethanoyl-12 is found from asclepiadaceae plant, 20-O-diisoamyl acyl-hardship rope glycoside and Cynanchum otophvllum glycoside derivative, and obtain new 11-O-ethanoyl-12 by hydrolysis, 20-O-diisoamyl acyl-hardship rope aglycon has stronger antiepileptic activity, and do not report this C-21 steroidal compounds and antiepileptic action thereof in document before this, the present invention is this compounds of isolation identification first, and by zebra fish anti-epileptic model validation C-3, 11, 12, it is effective to treatment epilepsy that 20-tetra-replaces C-21 steroid derivative.11-O-ethanoyl-12,20-O-diisoamyl acyl-hardship rope aglycon (11-O-acetyl-12,20-di-O-isovaleryldrevogenin) is C-21 steroid derivative, belongs to a kind of C-21 steroid compound of innovation.Zebra fish anti-epileptic model proves, it has stronger antiepileptic activity (IC
50be 17.2 μMs), be better than positive control drug Levetiracetam (IC far away
50be 170.6 μMs) and Cynanchum otophvllum sheet.And compared with Levetiracetam, Levetiracetam anti-epileptic therapeutic efficiency first increases fast, increase trend afterwards and slow down gradually, and this compound has the pharmacology that anti-epileptic therapeutic efficiency substantially linearly increases along with the increase of concentration.
Found by the model research of zebra fish anti-epileptic, C-3 of the present invention, 11,12,20-tetra-replaces the activity that C-21 steroid derivative and pharmacy acceptable salt and glycoside thereof have treatment epilepsy, significantly reduces attack degree and attack rate, in the concentration range of 1 μM to 100 μMs, anti-epileptic therapeutic efficiency increases along with the increase of concentration, presents concentration dependent.Compound 11-O-ethanoyl-12,20-O-diisoamyl acyl of the present invention-hardship rope aglycon, IC
50be 17.2, be significantly better than positive control Levetiracetam (IC
50be 170.6 μMs) and Cynanchum otophvllum sheet (52.8 μ g/ml).
C-3 of the present invention, 11,12,20-tetra-replaces C-21 steroid derivative and pharmaceutical composition can be any suitable form, such as solid, semi-solid, liquid or aerosol form.Generally, medicine contains compound of the present invention or extract as activeconstituents, with applicable outside, enteron aisle, or the organic or inorganic carrier of administered parenterally or mixed with excipients.Activeconstituents can be compound, such as, can accept carrier and/or vehicle make tablet, piller, capsule, suppository, vaginal suppository, solution, emulsion, suspension and be applicable to other forms of using with conventional non-toxic pharmaceutical.The pharmaceutical acceptable carrier used in the composition comprises, such as, water, glucose, lactose, gum arabic, gelatin, mannitol, starch, Magnesium Trisilicate, talcum, W-Gum, Keratin sulfate, colloidal silica, yam starch, and be adapted at preparing other carriers used in the preparation of solid, semisolid, liquid or aerosol form.Composition can contain stablizer, thickening material in addition, and/or tinting material and spices.
C-3 of the present invention, 11,12,20-tetra-replaces C-21 steroid derivative and pharmacy acceptable salt thereof and glycoside can per os or without mouth administration, dosage is had nothing in common with each other because medicine is different, and concerning adult, every day, 1-100mg was more suitable.
During oral administration administration, first make compound mix as vehicle, disintegrating agent, tamanori, lubricant, antioxidant, Drug coating, tinting material, perfume compound, tensio-active agent etc. with conventional medicinal adjuvant, be made into the form administrations such as granule, capsule, tablet; Can the form administration such as injection liquid, infusion solution or suppository during non-oral administration.When preparing above-mentioned preparation, conventional preparation technique can be used.
Accompanying drawing illustrates:
Fig. 1-1 is the zebra fish movement locus after the process of Compound D V12a each concentration group;
Fig. 1-2 is compound 11-O-ethanoyl-12, zebra fish rapid movement distance after 20-O-diisoamyl acyl-hardship rope aglycon each concentration group process, 11-O-ethanoyl-12,20-O-diisoamyl acyl-hardship restricts each concentration group of aglycon with phenytoin Sodium compared with epilepsy model group, p<0.001;
Fig. 1-3 is the dose-effect curve of compound 11-O-ethanoyl-12,20-O-diisoamyl acyl-hardship rope aglycon antiepileptic action;
Fig. 2-1 is the zebra fish movement locus after the process of Cynanchum otophvllum glycosides H each concentration group;
Fig. 2-2 is the zebra fish rapid movement distance after the process of Cynanchum otophvllum glycosides H each concentration group, each concentration group of Cynanchum otophvllum glycosides H with phenytoin Sodium compared with epilepsy model group, p<0.001;
Fig. 2-3 is the dose-effect curve of Cynanchum otophvllum glycosides H antiepileptic action.
Embodiment:
Below in conjunction with accompanying drawing, further illustrate essentiality content of the present invention with embodiments of the invention, these examples are only the explanations to preferred version of the present invention, and also limit the scope of the invention never in any form.
Embodiment 1:
The evaluation of antiepileptic action in 11-O-acetyl ester group-12,20-O-diisoamyl acyl ester group-hardship rope aglycon body:
11-O-ethanoyl-12; the preparation of 20-O-diisoamyl acyl-hardship rope aglycon: dry South Mountain rattan (5.5kg) is pulverized, and extracts three times, each four hours with methanol eddy; cooled and filtered; united extraction liquid, is concentrated into about 1.5L, after petroleum ether extraction removing fat-soluble component; reclaim methyl alcohol to medicinal extract shape; enriched material is suspended in water, with chloroform extraction four times, obtains chloroform extract 168g.Get 150g to dissolve, sample mixed by 300g silica gel (80-100 order), is separated, CHCl with silica gel column chromatography (200-300 order, 3270g)
3-MeOH-H
2o (86:17:1) wash-out, is mainly divided into three part Frs.A-C of opposed polarity.Fr.C (21g) is little polar portion, separating difficulty is relatively little, main utilization silica gel column chromatography is separated (chloroform/methanol or sherwood oil/acetone are eluent), Rp-8 reverse phase silica gel and MCI-gel CHP-20P column chromatography purification (methanol/water is eluent), obtain new compound root of Twisting Dregea glycosides I (97mg).Root of Twisting Dregea glycosides I is dissolved in MeOH (20mL), adds 5%HCl (10mL), after 50 ° of C water-bath 15min, add H
2o (20mL), reclaim under reduced pressure MeOH to 30mL, then after 60 ° of C water-bath 15min, cooling, extracts three times by equal amounts of chloroform, obtains glucoside unit part.Glucoside unit part, through silica gel column chromatography separating purification, with sherwood oil/acetone mixture wash-out, obtains 11-O-ethanoyl-12,20-O-diisoamyl acyl-hardship rope aglycon.
11-O-acetyl ester group-12,20-O-diisoamyl acyl ester group-hardship rope aglycon
11-O-acetyl-12,20-di-O-isovaleryldrevogenin
The physicochemical data of compound 11-O-acetyl-12,20-di-O-isovaleryldrevogenin is as follows: white amorphous powder, C
33h
52o
8; Mp 97-99 ° C,
(c=0.27, MeOH), IR (KBr) ν
max3441,2961,2934,2873,1742,1631,1467,1415,1370,1294,1254,1236,1196,1168,1120,1094,1061,1029,1000,963,940,901,811,771cm
-1.ESI-MS (positive ion) m/z 599 [M+Na]
+, HRESI-MS (positive ion) m/z 599.3568 [M (C
33h
52o
8)+Na]
+(calcd.599.3559).
1H-NMR(C
5D
5N,400MHz):δ1.46(1H,m,H-1a),2.08(1H,m,H-1b),1.82(1H,m,H-2a),2.10(1H,m,H-2b),3.85(1H,m,H-3),2.41(1H,m,H-4a),2.61(1H,m,H-4b),5.51(1H,d,J=4.9Hz,H-6),2.01(1H,m,H-7a),2.61(1H,m,H-7b),2.22(1H,m,H-8),1.86(1H,m,H-9),5.77(1H,t,J=9.8Hz,H-11),5.28(1H,d,J=9.8Hz,H-12),1.89(1H,m,H-15a),1.98(1H,m,H-15b),1.81(1H,m,H-16a),1.99(1H,m,H-16b),2.77(1H,m,H-17),1.46(3H,s,Me-18),1.31(3H,s,Me-19),5.47(1H,m,H-20),1.20(3H,d,J=5.9Hz,Me-21),2.12(3H,s,Me-2′),2.38(1H,m,H-2″a),2.51(1H,m,H-2″b),2.35(1H,m,H-3″),1.04(3H,d,J=6.4Hz,Me-4″),1.04(3H,d,J=6.4Hz,Me-5″),2.27(2H,m,H-2″′),2.20(1H,m,H-3″′),0.95(3H,d,J=6.4Hz,Me-4″′),0.95(3H,d,J=6.4Hz,Me-5″′)13C-NMR(C5D5N，100MHz)：δ39.0(t,C-1),32.8(t,C-2),70.9(d,C-3),43.9(t,C-4),140.8(s,C-5),121.9(d,C-6),28.2(t,C-7),38.0(d,C-8),48.2(d,C-9),39.6(s,C-10),72.2(d,C-11),78.8(d,C-12),53.0(s,C-13),84.2(s,C-14),33.0(t,C-15),25.0(t,C-16),50.2(d,C-17),11.3(q,C-18),19.5(q,C-19),73.2(d,C-20),19.5(q,C-21),170.2(s,C-1'),21.8(q,C-2'),173.2(s,C-1″),43.9(t,C-2″),25.7(d,C-3″),22.7(q,C-4″),22.8(q,C-5″),172.3(s,C-1″′),44.1(t,C-2″′),25.9(d,C-3″′),22.5(q,C-4″′),22.5(q,C-5″′)。
The breeding of zebrafish embryo is carried out in the mode of natural paired cross.Each mating prepares 4 ~ 5 pairs of Adult Zebrafishs, and on average often pair can be produced 200 ~ 300 embryos.At after fertilization 6 hours (i.e. 6hpf) and 24hpf, embryo is cleared up (removing dead embryo), and select suitable embryo according to the etap of embryo.Embryo's (fish culture water water quality: add 200mg Instant Ocean in every 1L reverse osmosis water, specific conductivity is 480 ~ 510 μ S/cm is hatched with fish culture water under 28 DEG C of conditions; PH is 6.9 ~ 7.2; Hardness is 53.7 ~ 71.6mg/LCaCO
3).Because embryo can obtain nutritive substance from the yolk sac of self, so (9dpf) does not need feeding in after fertilization 9 days.After having tested, with tricaine methylsulfonic acid, over-exposure process is carried out to the zebra fish of each etap, thus zebra fish anesthesia is put to death.The operation steps that anesthesia is put to death meets the code requirement that American Veterinary association (AVMA) puts to death Animal Anesthesia.
The zebra fish of Yetrazol process is epilepsy model group; The zebra fish of phenytoin Sodium and Yetrazol co-treatment is positive controls; The zebra fish of the DMSO process of 1.1% is negative control group (solvent control group); Untreated zebra fish is blank group.11-O-acetyl ester group-12,20-O-diisoamyl acyl ester group-hardship rope aglycon and positive control (Levetiracetam and Cynanchum otophvllum sheet) are dissolved in DMSO respectively, are mixed with the solution to be measured that concentration is 1,3,10,30,60 and 100 μM.Above-mentioned solution to be measured respectively with Yetrazol co-treatment zebra fish for some time after, utilize the movement locus of zebra fish in behavioural analysis instrument record 60min, then to the rapid movement (seizure of zebra fish, V>20mm/sec) distance carries out quantitative analysis, by calculating following index: 1. zebra fish rapid movement distance (distance ofrapid movement-seizure); 2. the anti-epileptic therapeutic efficiency (therapeutic efficacy) of compound; 3. compound time changing curve (compound effect on the time-course of zebrafish seizure) that zebra fish rapid movement is affected.Quantitative analysis is carried out, the anti-epileptic therapeutic efficiency of computerized compound with distance (D) to the movement velocity of zebra fish, and draws dose-effect curve according to compound epilepsy therapy efficiency, the IC of computerized compound antiepileptic action
50; Carry out statistical analysis with variance analysis and T-inspection, p<0.05 is significant difference.In experimentation all there are not the phenomena of mortality in each group zebra fish.Fig. 1-1 is each experimental group zebra fish movement locus in 60min, and restrict each concentration group of aglycon compared with epilepsy model group from Fig. 1-1,11-O-acetyl ester group-12,20-O-diisoamyl acyl ester group-hardship, zebra fish rapid movement track obviously reduces.Fig. 1-2 is the column diagram drawn according to zebra fish rapid movement distance average mean ± SE, and from Fig. 1-2, each concentration group all has statistical significance (p<0.001) compared with solvent control group; Fig. 1-3 is 11-O-acetyl ester group-12; the dose-effect curve of 20-O-diisoamyl acyl ester group-hardship rope aglycon antiepileptic action; from Fig. 1-3; 11-O-ethanoyl-12; 20-O-diisoamyl acyl-hardship rope aglycon is in the concentration range of 1 μM to 100 μMs; anti-epileptic therapeutic efficiency increases along with the increase of concentration, presents concentration dependent, IC
50it is 17.2 μMs.
Table 1.11-O-acetyl ester group-12,20-O-diisoamyl acyl ester group-hardship rope aglycon each concentration group zebra fish rapid movement distance raw data
Embodiment 2:
The evaluation of antiepileptic action in Cynanchum otophvllum glycosides H [Qingyanshengenin-3-O-β-D-glucopyanosyl base-(1 → 4)-β-D-glucopyanosyl base-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-Folium seu Cortex Nerii pyrans glycosyl-(1 → 4)-β-D-digitoxin pyranoside, Otophylloside H] body.
The preparation of Cynanchum otophvllum glycosides H [Qingyanshengenin-3-O-β-D-glucopyanosyl base-(1 → 4)-β-D-glucopyanosyl base-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-Folium seu Cortex Nerii pyrans glycosyl-(1 → 4)-β-D-digitoxin pyranoside, Otophylloside H]:
Dry Cynanchum otophvllum root (10.0kg) is pulverized, and extracts three times (72,72,48 hours), each consumption 15L by 90% ethanol soaking at room temperature, filters, united extraction liquid, and recycling design, to without alcohol taste, obtains fluid extract 368g.Be suspended in by medicinal extract in 2L water, with chloroform extraction three times, each consumption 1.5L, combining extraction liquid, is concentrated into dry, obtains chloroform extract 155g.Get 150g to be separated through silica gel column chromatography, chloroform/methanol (10:1.5,7.6L) wash-out, obtain five part Frs.1-5.Fr.3 is through silica gel column chromatography, with chloroform/methanol and ethyl acetate/ethanol/water for eluent, and Rp-8 column chromatography (taking methanol/water as eluent) separation and purification repeatedly, obtain Cynanchum otophvllum glycosides H [Qingyanshengenin-3-O-β-D-glucopyanosyl base-(1 → 4)-β-D-glucopyanosyl base-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-Folium seu Cortex Nerii pyrans glycosyl-(1 → 4)-β-D-digitoxin pyranoside, Otophylloside H].
Cynanchum otophvllum glucoside H, Otophylloside H
The physicochemical data of Cynanchum otophvllum glycosides H is as follows: white amorphous powder, C
60h
90o
27; Mp 213-216 ° C,
(c=0.83, MeOH), IR (KBr) ν
max3442,2934,1709,1610,1591,1504,1452,1384,1309,1276,1163,1072,911,854,773cm
-1.FAB-MS (negative ion) m/z 1242 [M]
-, 1105 [M-137]
-, 943 [M-137-162]
-, HRFAB-MS (negative ion) m/z1241.5628 [M (C
60h
90o
27)-H]
-(calcd.1241.5591).
1H-NMR(C
5D
5N,500MHz):δ3.54(1H,m,H-3),5.38(1H,d,J=4.7Hz,H-6),4.72(1H,dd,J=11.6Hz,J=4.1Hz,H-12),1.63(3H,s,Me-18),1.14(3H,s,Me-19),2.08(3H,s,Me-21),7.73(1H,d,J=8.8Hz,H-3'),6.71(1H,d,J=8.8Hz,H-4'),6.71(1H,d,J=8.8Hz,H-6'),7.73(1H,d,J=8.8Hz,H-7'),4.94(1H,dd,J=9.7Hz,J=1.7Hz,H-1″),1.67(1H,m,H-2″a),1.93(1H,m,H-2″b),4.21(1H,m,H-3″),3.27(1H,m,H-4″),3.80(1H,m,H-5″),1.21(3H,d,J=5.8Hz,H-6″),4.60(dd,J=9.6Hz,J=1.6Hz,H-1″′),1.41(1H,m,H-2a″′),2.34(1H,m,H-2b″′),3.41(1H,m,H-3″′),3.18(1H,m,H-4″′),3.40(1H,m,H-5″′),1.38(3H,d,J=6.2Hz,H-6″′),3.48(3H,s,Me-3″′),4.82(1H,br d,J=9.7Hz,H-1″″),1.73(1H,m,H-2a″″),2.45(1H,m,H-2b″″),3.55(1H,m,H-3″″),3.51(1H,m,H-4″″),3.52(1H,m,H-5″″),1.43(3H,d,J=6.4Hz,H-6″″),3.51(3H,s,Me-3″″),4.48(1H,d,J=7.9Hz,H-1″″′),3.23(1H,m,H-2″″′),3.29(1H,m,H-3″″′),3.52(1H,m,H-4″″′),3.51(1H,m,H-5″″′),3.82(1H,m,H-6a″″′),3.92(1H,dd,J=12.0Hz,J=2.2Hz,H-6b″″′),4.38(1H,d,J=7.8Hz,H-1″″″),3.21(1H,m,H-2″″″),3.34(1H,m,H-3″″″),3.28(1H,m,H-4″″″),3.32(1H,m,H-5″″″),3.64(1H,dd,J=12.0Hz,J=5.8Hz,H-6a″″″),3.87(1H,dd,J=12.0Hz,J=2.2Hz,H-6b″″″)。
13C-NMR(C
5D
5N,100MHz):δ39.8(t,C-1),30.1(t,C-2),79.3(d,C-3),39.8(t,C-4),140.3(s,C-5),119.4(d,C-6),35.2(t,C-7),75.0(s,C-8),45.1(d,C-9),38.1(s,C-10),25.5(t,C-11),73.7(d,C-12),59.1(s,C-13),90.0(s,C-14),34.3(t,C-15),33.4(t,C-16),93.1(s,C-17),10.6(q,C-18),18.6(q,C-19),211.9(s,C-20),27.7(q,Me-21),167.3(s,C-1'),119.6(s,C-2'),132.8(d,C-3'),117.6(d,C-4'),168.3(s,C-5'),117.6(d,C-6'),132.8(d,C-7'),97.0(d,C-1″),38.8(t,C-2″),68.3(d,C-3″),83.7(d,C-4″),69.5(d,C-5″),18.5(q,C-6″),102.6(d,C-1″′),37.7(t,C-2″′),80.1(d,C-3″′),83.6(d,C-4″′),72.6(d,C-5″′),18.8(q,C-6″′),57.9(q,Me-3″′),100.6(d,C-1″″),36.3(t,C-2″″),78.4(d,C-3″″),83.6(d,C-4″″),70.0(d,C-5″″),18.6(q,C-6″″),58.6(q,Me-3″″),104.0(d,C-1″″′),75.3(d,C-2″″′),76.7(d,C-3″″′),81.0(d,C-4″″′),76.4(d,C-5″″′),62.1(t,C-6″″′),104.6(d,C-1″″″),74.9(d,C-2″″″),78.1(d,C-3″″″),71.4(d,C-4″″″),77.8(d,C-5″″″),62.4(t,C-6″″″)。
The breeding of zebrafish embryo is carried out in the mode of natural paired cross.Each mating prepares 4 ~ 5 pairs of Adult Zebrafishs, and on average often pair can be produced 200 ~ 300 embryos.At after fertilization 6 hours (i.e. 6hpf) and 24hpf, embryo is cleared up (removing dead embryo), and select suitable embryo according to the etap of embryo.Embryo's (fish culture water water quality: add 200mg Instant Ocean in every 1L reverse osmosis water, specific conductivity is 480 ~ 510 μ S/cm is hatched with fish culture water under 28 DEG C of conditions; PH is 6.9 ~ 7.2; Hardness is 53.7 ~ 71.6mg/LCaCO3).Because embryo can obtain nutritive substance from the yolk sac of self, so (9dpf) does not need feeding in after fertilization 9 days.After having tested, with tricaine methylsulfonic acid, over-exposure process is carried out to the zebra fish of each etap, thus zebra fish anesthesia is put to death.The operation steps that anesthesia is put to death meets the code requirement that American Veterinary association (AVMA) puts to death Animal Anesthesia.
The zebra fish of Yetrazol process is epilepsy model group; The zebra fish of phenytoin Sodium and Yetrazol co-treatment is positive controls; The zebra fish of the DMSO process of 1.1% is negative control group (solvent control group); Untreated zebra fish is blank group.Cynanchum otophvllum glycosides H and positive control (Levetiracetam and Cynanchum otophvllum sheet) are dissolved in DMSO respectively, are mixed with the solution to be measured that concentration is 1,3,10,30,60,100,150 and 300 μM.Above-mentioned solution to be measured respectively with Yetrazol co-treatment zebra fish certain hour, utilize the movement locus of zebra fish in behavioural analysis instrument record 60min, then to the rapid movement (seizure of zebra fish, V>20mm/sec) distance carries out quantitative analysis, by calculating following index: 1. zebra fish rapid movement distance (distance of rapid movement-seizure); 2. the anti-epileptic therapeutic efficiency (therapeuticefficacy) of compound; 3. compound time changing curve (compound effect onthe time-course of zebrafish seizure) that zebra fish rapid movement is affected.Quantitative analysis is carried out, the anti-epileptic therapeutic efficiency of computerized compound with distance (D) to the movement velocity of zebra fish, and draws dose-effect curve according to compound epilepsy therapy efficiency, the IC of computerized compound antiepileptic action
50; Carry out statistical analysis with variance analysis and T-inspection, p<0.05 is significant difference.In experimentation all there are not the phenomena of mortality in each group zebra fish.Fig. 2-1 is each experimental group zebra fish movement locus in 60min, and from Fig. 2-1, each concentration group of compound Cynanchum otophvllum glycosides H is compared with epilepsy model group, and zebra fish rapid movement track obviously reduces.Fig. 2-2 is the column diagram drawn according to zebra fish rapid movement distance average mean ± SE, and from Fig. 2-2, each concentration group all has statistical significance (p<0.001) compared with solvent control group; Fig. 2-3 is the dose-effect curve of compound Cynanchum otophvllum glycosides H antiepileptic action, from Fig. 2-3, compound Cynanchum otophvllum glycosides H is in the concentration range of 1 μM to 300 μMs, anti-epileptic therapeutic efficiency increases along with the increase of concentration, present concentration dependent, when concentration is 30 μMs, anti-epileptic therapeutic efficiency is 45% ± 2.13%.
Table 2. Cynanchum otophvllum glycosides H each concentration group zebra fish rapid movement distance raw data
Embodiment 3:
Root of Twisting Dregea glycosides Da
1(Dregeoside Da
1) evaluation of antiepileptic action in body:
Root of Twisting Dregea glycosides Da
1the preparation of (Dregeoside Da1): dry South Mountain rattan (5.5kg) is pulverized, three times are extracted with methanol eddy, each four hours, cooled and filtered, united extraction liquid, be concentrated into about 1.5L, after petroleum ether extraction removing fat-soluble component, reclaim methyl alcohol to medicinal extract shape, enriched material is suspended in water, with chloroform extraction four times, obtain chloroform extract 168g.Get 150g to dissolve, sample mixed by 300g silica gel (80-100 order), is separated, CHCl with silica gel column chromatography (200-300 order, 3270g)
3-MeOH-H
2o (86:17:1) wash-out, is mainly divided into three part Frs.A-C of opposed polarity.Fr.C (21g) is little polar portion, separating difficulty is relatively little, main utilization silica gel column chromatography is separated (chloroform/methanol or sherwood oil/acetone are eluent), Rp-8 reverse phase silica gel and MCI-gel CHP-20P column chromatography purification (methanol/water is eluent), obtain compound root of Twisting Dregea glycosides Da
1(46mg).
Root of Twisting Dregea glycosides Da
1, Dregeoside Da
1
The breeding of zebrafish embryo is carried out in the mode of natural paired cross.Each mating prepares 4 ~ 5 pairs of Adult Zebrafishs, and on average often pair can be produced 200 ~ 300 embryos.At after fertilization 6 hours (i.e. 6hpf) and 24hpf, embryo is cleared up (removing dead embryo), and select suitable embryo according to the etap of embryo.Embryo's (fish culture water water quality: add 200mg Instant Ocean in every 1L reverse osmosis water, specific conductivity is 480 ~ 510 μ S/cm is hatched with fish culture water under 28 DEG C of conditions; PH is 6.9 ~ 7.2; Hardness is 53.7 ~ 71.6mg/LCaCO
3).Because embryo can obtain nutritive substance from the yolk sac of self, so (9dpf) does not need feeding in after fertilization 9 days.After having tested, with tricaine methylsulfonic acid, over-exposure process is carried out to the zebra fish of each etap, thus zebra fish anesthesia is put to death.The operation steps that anesthesia is put to death meets the code requirement that American Veterinary association (AVMA) puts to death Animal Anesthesia.
The zebra fish of Yetrazol process is epilepsy model group; The zebra fish of phenytoin Sodium and Yetrazol co-treatment is positive controls; The zebra fish of the DMSO process of 1.1% is negative control group (solvent control group); Untreated zebra fish is blank group.Root of Twisting Dregea glycosides Da1 is dissolved in DMSO, is mixed with the solution to be measured that concentration is 30 μMs.Above-mentioned solution to be measured respectively with Yetrazol co-treatment zebra fish certain hour, utilize the movement locus of zebra fish in behavioural analysis instrument record 60min, then to the rapid movement (seizure of zebra fish, V>20mm/sec) distance carries out quantitative analysis, by calculating following index: 1. zebra fish rapid movement distance (distance of rapid movement-seizure); 2. the anti-epileptic therapeutic efficiency (therapeutic efficacy) of compound; 3. compound time changing curve (compound effect on the time-course of zebrafish seizure) that zebra fish rapid movement is affected.Root of Twisting Dregea glycosides Da1 and phenytoin Sodium are compared with epilepsy model group, and zebra fish rapid movement track obviously reduces, and preliminary deterministic compound root of Twisting Dregea glycosides Da1 has antiepileptic action, and therapeutic efficiency is 40 ± 3.13%.
Embodiment 4:
Root of Twisting Dregea glycosides D [11,12-di-O-acetyldrevogenin P-3-O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyranoside, Drevoluoside D] evaluation of antiepileptic action in body
Root of Twisting Dregea glycosides D [11,12-di-O-acetyldrevogenin P-3-O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyranoside, Drevoluoside D] preparation:
Dry South Mountain rattan (5.5kg) is pulverized, three times are extracted with methanol eddy, each four hours, cooled and filtered, united extraction liquid, be concentrated into about 1.5L, after petroleum ether extraction removing fat-soluble component, reclaim methyl alcohol to medicinal extract shape, enriched material is suspended in water, with chloroform extraction four times, obtain chloroform extract 168g.Get 150g to dissolve, sample mixed by 300g silica gel (80-100 order), is separated, CHCl with silica gel column chromatography (200-300 order, 3270g)
3-MeOH-H
2o (86:17:1) wash-out, is mainly divided into three part Frs.A-C of opposed polarity.Fr.C (21g) is little polar portion, separating difficulty is relatively little, main utilization silica gel column chromatography is separated (chloroform/methanol or sherwood oil/acetone are eluent), Rp-8 reverse phase silica gel and MCI-gel CHP-20P column chromatography purification (methanol/water is eluent), obtain root of Twisting Dregea glycosides D (8mg).
Root of Twisting Dregea glycosides D, Drevoluoside D
The physicochemical data of root of Twisting Dregea glycosides D is as follows: white amorphous powder, C
46h
72o
17; Mp 116-120 ° C,
(c=0.20, MeOH), IR (KBr) ν
max3442,2971,2932,2856,1748,1700,1612,1450,1369,1318,1247,1226,1195,1166,1127,1084,1030,1004,964,915,866,816,785,721cm
-1.FAB-MS (negative ion) m/z 895 [M-1]
-, HRESI-MS (negative ion) m/z 931.4434 [M (C
46h
72o
17)+Cl]
-(calcd.931.4458).
1H-NMR(C
5D
5N,500MHz):δ1.33(1H,m,H-1a),1.98(1H,m,H-1b),1.74(1H,m,H-2a),2.10(1H,m,H-2b),3.77(1H,m,H-3),2.30(1H,m,H-4a),2.50(1H,m,H-4b),5.43(1H,d,J=5.4Hz,H-6),1.94(1H,m,H-7a),2.58(1H,m,H-7b),2.03(1H,m,H-8),3.08(1H,m,H-9),5.65(1H,t,J=10.3Hz,H-11),5.12(1H,d,J=10.3Hz,H-12),1.95(1H,m,H-15a),2.05(1H,m,H-15b),1.96(1H,m,H-16a),2.29(1H,m,H-16b),3.08(1H,m,H-17),1.30(3H,s,Me-18),1.19(3H,s,Me-19),2.20(3H,s,Me-21),2.02(3H,s,Me-2'),2.18(3H,s,Me-2″),5.23(1H,br d,J=9.3Hz,cym-1),1.85(1H,m,cym-2a),2.27(1H,m,cym-2b),4.03(1H,m,cym-3),3.44(1H,dd,J=9.8Hz,J=2.4Hz,cym-4),4.19(1H,m,cym-5),1.33(3H,d,J=6.4Hz,cym-6),3.57(3H,s,Me-cym-3),5.09(1H,br d,J=9.3Hz,cym-1'),1.82(1H,m,cym-2a'),2.33(1H,m,cym-2b'),4.08(1H,m,cym-3'),3.53(1H,dd,J=9.8Hz,J=2.4Hz,cym-4'),4.18(1H,m,cym-5'),1.49(3H,d,J=6.4Hz,cym-6'),3.58(3H,s,Me-cym-3'),5.12(1H,d,J=7.8Hz,alm-1),3.88(1H,m,alm-2),4.07(1H,m,alm-3),3.60(1H,br d,J=9.8Hz,alm-4),4.15(1H,m,alm-5),1.53(3H,d,J=6.4Hz,cym-6'),3.84(3H,s,Me-alm-3')。
13C-NMR(C
5D
5N,125MHz)：δ38.8(t,C-1),30.5(t,C-2),77.2(d,C-3),39.8(t,C-4),139.7(s,C-5),122.6(d,C-6),28.3(t,C-7),37.4(d,C-8),47.9(d,C-9),39.5(s,C-10),71.9(d,C-11),78.0(d,C-12),54.6(s,C-13),84.0(s,C-14),34.5(t,C-15),24.0(t,C-16),58.3(d,C-17),11.5(q,C-18),19.3(q,C-19),213.6(s,C-20),31.8(q,Me-21),170.2(s,C-1'),21.4(q,C-2'),171.1(s,C-1″),20.8(q,C-2″),96.5(d,cym-1),37.3(t,cym-2),78.0(d,cym-3),83.3(d,cym-4),69.1(d,cym-5),18.6(q,cym-6),58.8(q,Me-cym-3),100.4(d,cym-1'),36.9(t,cym-2'),78.2(d,cym-3'),83.3(d,cym-4'),69.4(d,cym-5'),18.6(q,cym-6'),58.8(q,Me-cym-3'),104.2(d,alm-1),73.1(d,alm-2),84.0(d,alm-3),74.5(d,alm-4),70.8(d,alm-5),18.6(d,alm-6),62.2(q,Me-alm-3')。
The breeding of zebrafish embryo is carried out in the mode of natural paired cross.Each mating prepares 4 ~ 5 pairs of Adult Zebrafishs, and on average often pair can be produced 200 ~ 300 embryos.At after fertilization 6 hours (i.e. 6hpf) and 24hpf, embryo is cleared up (removing dead embryo), and select suitable embryo according to the etap of embryo.Embryo's (fish culture water water quality: add 200mg Instant Ocean in every 1L reverse osmosis water, specific conductivity is 480 ~ 510 μ S/cm is hatched with fish culture water under 28 DEG C of conditions; PH is 6.9 ~ 7.2; Hardness is 53.7 ~ 71.6mg/LCaCO
3).Because embryo can obtain nutritive substance from the yolk sac of self, so (9dpf) does not need feeding in after fertilization 9 days.After having tested, with tricaine methylsulfonic acid, over-exposure process is carried out to the zebra fish of each etap, thus zebra fish anesthesia is put to death.The operation steps that anesthesia is put to death meets the code requirement that American Veterinary association (AVMA) puts to death Animal Anesthesia.
The zebra fish of Yetrazol process is epilepsy model group; The zebra fish of phenytoin Sodium and Yetrazol co-treatment is positive controls; The zebra fish of the DMSO process of 1.1% is negative control group (solvent control group); Untreated zebra fish is blank group.Root of Twisting Dregea glycosides D is dissolved in DMSO, is mixed with the solution to be measured that concentration is 30 μMs.Above-mentioned solution to be measured respectively with Yetrazol co-treatment zebra fish certain hour, utilize the movement locus of zebra fish in behavioural analysis instrument record 60min, then to the rapid movement (seizure of zebra fish, V>20mm/sec) distance carries out quantitative analysis, by calculating following index: 1. zebra fish rapid movement distance (distance of rapid movement-seizure); 2. the anti-epileptic therapeutic efficiency (therapeutic efficacy) of compound; 3. compound time changing curve (compound effect on the time-course of zebrafish seizure) that zebra fish rapid movement is affected.Root of Twisting Dregea glycosides D and phenytoin Sodium are compared with epilepsy model group, and zebra fish rapid movement track obviously reduces, and preliminary deterministic compound root of Twisting Dregea glycosides D has antiepileptic action, and therapeutic efficiency is 38 ± 4.43%.
Embodiment 5:
The evaluation of antiepileptic action in Cynanchum otophvllum glycosides N [Qingyanshengenin-3-O-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-Folium seu Cortex Nerii pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-digitoxin pyranoside, Otophylloside N] body
The preparation of Cynanchum otophvllum glycosides N [Qingyanshengenin-3-O-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-Folium seu Cortex Nerii pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-digitoxin pyranoside, Otophylloside N]:
Dry Cynanchum otophvllum root (10.0kg) is pulverized, and extracts three times (72,72,48 hours), each consumption 15L by 90% ethanol soaking at room temperature, filters, united extraction liquid, and recycling design, to without alcohol taste, obtains fluid extract 368g.Be suspended in by medicinal extract in 2L water, with chloroform extraction three times, each consumption 1.5L, combining extraction liquid, is concentrated into dry, obtains chloroform extract 155g.Get 150g to be separated through silica gel column chromatography, chloroform/methanol (10:1.5,7.6L) wash-out, obtain five part Frs.1-5.Fr.1 is separated repeatedly through silica gel column chromatography, with chloroform/methanol and sherwood oil/acetone for eluent, obtains Cynanchum otophvllum glycosides N (10mg).
Cynanchum otophvllum glycosides N, Otophylloside N
The physicochemical data of Cynanchum otophvllum glucoside N (Otophylloside N): white amorphous powder, C
55h
82o
20; Mp163-165 ° of C,
(c=0.20, MeOH), IR (KBr) ν
max3446,2971,2932,1713,1610,1594,1516,1452,1382,1369,1308,1275,1164,1093,1004,912,866,852,772cm
-1.FAB-MS (negative ion) m/z 1061 [M-1]
-, HRESI-MS (negative ion) m/z 1061.5284 [M (C
55h
82o
20)-H]
-(calcd.1061.5321).
1H-NMR(C
5D
5N,500MHz):δ3.87(1H,m,H-3),5.26(1H,br s,H-6),5.33(1H,dd,J=11.5Hz,J=4.0Hz,H-12),2.09(3H,s,Me-18),1.30(3H,s,Me-19),2.41(3H,s,Me-21),8.30(1H,d,J=8.6Hz,H-3'),7.23(1H,d,J=8.6Hz,H-4'),7.23(1H,d,J=8.6Hz,H-6'),8.30(1H,d,J=8.6Hz,H-7'),5.48(1H,br d,J=9.2Hz,H-1″),2.07(1H,m,H-2″a),2.40(1H,m,H-2″b),4.64(1H,m,H-3″),3.52(1H,m,H-4″),4.30(1H,m,H-5″),1.43(3H,d,J=6.4Hz,H-6″),5.17(1H,br d,J=9.8Hz,H-1″′),1.78(1H,m,H-2a″′),2.29(1H,m,H-2b″′),4.01(1H,m,H-3″′),3.41(1H,m,H-4″′),4.18(1H,m,H-5″′),1.32(3H,d,J=6.1Hz,H-6″′),3.56(3H,s,Me-3″′),4.68(1H,br d,J=9.7Hz,H-1″″),1.73(1H,m,H-2a″″),2.45(1H,m,H-2b″″),3.55(1H,m,H-3″″),3.51(1H,m,H-4″″),3.52(1H,m,H-5″″),1.43(3H,d,J=6.4Hz,H-6″″),3.51(3H,s,Me-3″″),5.25(1H,br d,J=9.3Hz,H-1″″′),1.77(1H,m,H-2a″″′),2.38(1H,m,H-2b″″′),3.77(1H,m,H-3″″′),3.54(1H,m,H-4″″′),4.14(1H,m,H-5″″′),1.54(3H,d,J=6.3Hz,H-6″″′),3.46(3H,s,Me-3″″′)。
13C-NMR(C
5D
5N,100MHz)：δ39.3(t,C-1),29.9(t,C-2),77.6(d,C-3),38.9(t,C-4),139.4(s,C-5),119.2(d,C-6),34.8(t,C-7),74.3(s,C-8),44.5(d,C-9),37.4(s,C-10),25.2(t,C-11),73.4(d,C-12),58.4(s,C-13),89.6(s,C-14),33.9(t,C-15),33.2(t,C-16),92.5(s,C-17),10.9(q,C-18),18.2(q,C-19),209.9(s,C-20),27.9(q,Me-21),165.4(s,C-1'),122.0(s,C-2'),132.5(d,C-3′),116.2(d,C-4'),163.7(s,C-5'),116.2(d,C-6'),132.5(d,C-7'),96.4(d,C-1″),39.1(t,C-2″),67.6(d,C-3″),83.4(d,C-4″),68.6(d,C-5″),18.7(q,C-6″),99.8(d,C-1″′),36.7(t,C-2″′),77.7(d,C-3″′),83.2(d,C-4″′),69.1(d,C-5″′),18.5(q,C-6″′),58.9(q,Me-3″′),102.0(d,C-1″″),37.7(t,C-2″″),78.9(d,C-3″″),82.7(d,C-4″″),71.8(d,C-5″″),18.7(q,C-6″″),57.4(q,Me-3″″),98.6(d,C-1″″′),35.9(t,C-2″″′),79.0(d,C-3″″′),74.2(d,C-4″″′),71.3(d,C-5″″′),19.1(q,C-6″″′),58.1(q,Me-3″″′)。
The breeding of zebrafish embryo is carried out in the mode of natural paired cross.Each mating prepares 4 ~ 5 pairs of Adult Zebrafishs, and on average often pair can be produced 200 ~ 300 embryos.At after fertilization 6 hours (i.e. 6hpf) and 24hpf, embryo is cleared up (removing dead embryo), and select suitable embryo according to the etap of embryo.Embryo's (fish culture water water quality: add 200mg Instant Ocean in every 1L reverse osmosis water, specific conductivity is 480 ~ 510 μ S/cm is hatched with fish culture water under 28 DEG C of conditions; PH is 6.9 ~ 7.2; Hardness is 53.7 ~ 71.6mg/LCaCO
3).Because embryo can obtain nutritive substance from the yolk sac of self, so (9dpf) does not need feeding in after fertilization 9 days.After having tested, with tricaine methylsulfonic acid, over-exposure process is carried out to the zebra fish of each etap, thus zebra fish anesthesia is put to death.The operation steps that anesthesia is put to death meets the code requirement that American Veterinary association (AVMA) puts to death Animal Anesthesia.
The zebra fish of Yetrazol process is epilepsy model group; The zebra fish of phenytoin Sodium and Yetrazol co-treatment is positive controls; The zebra fish of the DMSO process of 1.1% is negative control group (solvent control group); Untreated zebra fish is blank group.Cynanchum otophvllum glycosides N is dissolved in DMSO, is mixed with the solution to be measured that concentration is 30 μMs.Above-mentioned solution to be measured respectively with Yetrazol co-treatment zebra fish certain hour, utilize the movement locus of zebra fish in behavioural analysis instrument record 60min, then to the rapid movement (seizure of zebra fish, V>20mm/sec) distance carries out quantitative analysis, by calculating following index: 1. zebra fish rapid movement distance (distance of rapid movement-seizure); 2. the anti-epileptic therapeutic efficiency (therapeutic efficacy) of compound; 3. compound time changing curve (compound effect on the time-course of zebrafish seizure) that zebra fish rapid movement is affected.Cynanchum otophvllum glycosides N and phenytoin Sodium are compared with epilepsy model group, and zebra fish rapid movement track obviously reduces, and preliminary deterministic compound Cynanchum otophvllum glycosides N has antiepileptic action, and therapeutic efficiency is 32 ± 4.50%.
Embodiment 6:
The evaluation of antiepileptic action in 11-O-acetyl-12-O-angeloyl-17 β-marsdenin-3-O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyranoside body
The preparation of 11-O-acetyl-12-O-angeloyl-17 β-marsdenin-3-O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyranoside:
Dry South Mountain rattan (5.5kg) is pulverized, three times are extracted with methanol eddy, each four hours, cooled and filtered, united extraction liquid, be concentrated into about 1.5L, after petroleum ether extraction removing fat-soluble component, reclaim methyl alcohol to medicinal extract shape, enriched material is suspended in water, with chloroform extraction four times, obtain chloroform extract 168g.Get 150g to dissolve, sample mixed by 300g silica gel (80-100 order), and be separated with silica gel column chromatography (200-300 order, 3270g), CHCl3-MeOH-H2O (86:17:1) wash-out, is mainly divided into three part Frs.A-C of opposed polarity.Fr.C (21g) is little polar portion, separating difficulty is relatively little, main utilization silica gel column chromatography is separated (chloroform/methanol or sherwood oil/acetone are eluent), Rp-8 reverse phase silica gel and MCI-gel CHP-20P column chromatography purification (methanol/water is eluent), obtain 11-O-acetyl-12-O-angeloyl-17 β-marsdenin-3-O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyranoside (3mg).
The physicochemical data of 11-O-acetyl-12-O-angeloyl-17 β-marsdenin-3-O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyranoside: white amorphous powder, C
49h
76o
18; FAB-MS (negative ion) m/z 951 [M-1]
-, 869 [M-83]
-.
1H-NMR(C
5D
5N,500MHz):δ2.50(1H,m,H-1a),2.56(1H,m,H-1b),1.92(1H,m,H-2a),2.15(1H,m,H-2b),3.87(1H,m,H-3),1.34(1H,m,H-4a),2.36(1H,m,H-4b),5.33(1H,d,J=5.6Hz,H-6),2.05(1H,m,H-7a),2.22(1H,m,H-7b),2.14(1H,m,H-9),6.27(1H,t,J=10.7Hz,H-11),5.42(1H,d,J=10.7Hz,H-12),2.30(1H,m,H-15a),2.59(1H,m,H-15b),2.03(1H,m,H-16a),2.29(1H,m,H-16b),3.15(1H,m,H-17),1.63(3H,s,Me-18),1.59(3H,s,Me-19),2.18(3H,s,Me-21),1.59(3H,s,Me-2'),7.19(1H,H-3″),1.65(3H,d,J=6.8Hz,Me-4″),1.97(3H,s,Me-5″),5.27(1H,br d,J=9.4Hz,cym-1),1.85(1H,m,cym-2a),2.26(1H,m,cym-2b),4.02(1H,m,cym-3),3.44(1H,dd,J=9.4Hz,J=2.6Hz,cym-4),4.19(1H,m,cym-5),1.34(3H,d,J=6.0Hz,cym-6),3.57(3H,s,Me-cym-3),5.09(1H,br d,J=9.4Hz,cym-1'),1.81(1H,m,cym-2a'),2.33(1H,m,cym-2b'),4.07(1H,m,cym-3'),3.52(1H,dd,J=9.4Hz,J=2.6Hz,cym-4'),4.17(1H,m,cym-5'),1.48(3H,d,J=6.0Hz,cym-6'),3.57(3H,s,Me-cym-3'),5.13(1H,d,J=8.1Hz,alm-1),3.87(1H,m,alm-2),4.07(1H,m,alm-3),3.59(1H,m,alm-4),4.16(1H,m,alm-5),1.53(3H,d,J=6.4Hz,cym-6'),3.85(3H,s,Me-alm-3')。
13C-NMR(C
5D
5N,125MHz)：δ39.8(t,C-1),30.2(t,C-2),77.8(d,C-3),40.5(t,C-4),139.7(s,C-5),118.8(d,C-6),36.8(t,C-7),76.0(s,C-8),49.1(d,C-9),39.3(s,C-10),71.7(d,C-11),78.4(d,C-12),55.7(s,C-13),85.6(s,C-14),35.5(t,C-15),24.4(t,C-16),59.5(d,C-17),13.7(q,C-18),18.1(q,C-19),213.3(s,C-20),31.4(q,Me-21),169.9(s,C-1'),21.5(q,C-2'),168.0(s,C-1″),128.6(t,C-2″),139.1(d,C-3″),14.4(q,C-4″),12.2(q,C-5″),96.5(d,cym-1),37.3(t,cym-2),78.0(d,cym-3),83.4(d,cym-4),69.1(d,cym-5),18.6(q,cym-6),58.8(q,Me-cym-3),100.4(d,cym-1'),36.9(t,cym-2'),78.2(d,cym-3'),83.3(d,cym-4'),69.4(d,cym-5'),18.6(q,cym-6'),58.8(q,Me-cym-3'),104.2(d,alm-1),73.2(d,alm-2),84.0(d,alm-3),74.5(d,alm-4),70.8(d,alm-5),18.6(d,alm-6),62.2(q,Me-alm-3')。
11-O-acetyl-12-O-angeloyl-17 β-marsdenin-3-O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyranoside
The breeding of zebrafish embryo is carried out in the mode of natural paired cross.Each mating prepares 4 ~ 5 pairs of Adult Zebrafishs, and on average often pair can be produced 200 ~ 300 embryos.At after fertilization 6 hours (i.e. 6hpf) and 24hpf, embryo is cleared up (removing dead embryo), and select suitable embryo according to the etap of embryo.Embryo's (fish culture water water quality: add 200mg Instant Ocean in every 1L reverse osmosis water, specific conductivity is 480 ~ 510 μ S/cm is hatched with fish culture water under 28 DEG C of conditions; PH is 6.9 ~ 7.2; Hardness is 53.7 ~ 71.6mg/LCaCO
3).Because embryo can obtain nutritive substance from the yolk sac of self, so (9dpf) does not need feeding in after fertilization 9 days.After having tested, with tricaine methylsulfonic acid, over-exposure process is carried out to the zebra fish of each etap, thus zebra fish anesthesia is put to death.The operation steps that anesthesia is put to death meets the code requirement that American Veterinary association (AVMA) puts to death Animal Anesthesia.
The zebra fish of Yetrazol process is epilepsy model group; The zebra fish of phenytoin Sodium and Yetrazol co-treatment is positive controls; The zebra fish of the DMSO process of 1.1% is negative control group (solvent control group); Untreated zebra fish is blank group.Glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyranoside is dissolved in DMSO 11-O-acetyl-12-O-angeloyl-17 β-marsdenin-3-O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans, is mixed with the solution to be measured that concentration is 30 μMs.Above-mentioned solution to be measured respectively with Yetrazol co-treatment zebra fish certain hour, utilize the movement locus of zebra fish in behavioural analysis instrument record 60min, then to the rapid movement (seizure of zebra fish, V>20mm/sec) distance carries out quantitative analysis, by calculating following index: 1. zebra fish rapid movement distance (distance of rapidmovement-seizure); 2. the anti-epileptic therapeutic efficiency (therapeutic efficacy) of compound; 3. compound time changing curve (compound effect on the time-course ofzebrafish seizure) that zebra fish rapid movement is affected.11-O-acetyl-12-O-angeloyl-17 β-marsdenin-3-O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyranoside with phenytoin Sodium compared with epilepsy model group, zebra fish rapid movement track obviously reduces, preliminary deterministic compound 11-O-acetyl-12-O-angeloyl-17 β-marsdenin-3-O-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyranoside has antiepileptic action, therapeutic efficiency is 24 ± 4.32%.
Embodiment 7:
The evaluation of antiepileptic action in root of Twisting Dregea glycosides N [Marsectohexol-3-O-β-D-glucopyanosyl base-(1 → 4)-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyranoside, Drevoluoside N] body
The preparation of root of Twisting Dregea glycosides N [Marsectohexol-3-O-β-D-glucopyanosyl base-(1 → 4)-6-deoxidation-3-O-methyl-β-D-A Luo pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyrans glycosyl-(1 → 4)-β-D-apocynum cannabinum pyranoside, Drevoluoside N]:
Dry South Mountain rattan (5.5kg) is pulverized, three times are extracted with methanol eddy, each four hours, cooled and filtered, united extraction liquid, be concentrated into about 1.5L, after petroleum ether extraction removing fat-soluble component, reclaim methyl alcohol to medicinal extract shape, enriched material is suspended in water, with chloroform extraction four times, obtain chloroform extract 168g.Get 150g to dissolve, sample mixed by 300g silica gel (80-100 order), is separated, CHCl with silica gel column chromatography (200-300 order, 3270g)
3-MeOH-H
2o (86:17:1) wash-out, is mainly divided into three part Frs.A-C of opposed polarity.Fr.A (27g) is large polar portion, be separated after (methanol/water is eluent) through Rp-8 reverse phase silica gel, recycle silicon glue (chloroform/methanol is eluent) and MCI-gel CHP-20P (methanol/water is eluent) column chromatography purification, obtain compound root of Twisting Dregea glycosides N (6mg).
Root of Twisting Dregea glycosides N, Drevoluoside N
The physicochemical data of root of Twisting Dregea glucoside N (Drevoluoside N): white amorphous powder, C
48h
80o
21; Mp 177-179 ° C,
(c=0.22, MeOH), IR (KBr) ν
max3442,3433,2970,2933,1637,1452,1402,1378,1371,1319,1262,1235,1196,1162,1080,1004,953,909,864,816,780,720cm
-1.FAB-MS (negative ion) m/z 991 [M-1]
-, HRESI-MS (negative ion) m/z 1027.4870 [M (C
48h
80o
21)+Cl]
-(calcd.1027.4880).
1H-NMR(C
5D
5N,500MHz):δ2.59(2H,m,H-1),2.02(1H,m,H-2a),2.13(1H,m,H-2b),3.92(1H,m,H-3),1.38(1H,m,H-4a),3.33(1H,m,H-4b),5.35(1H,d,J=4.9Hz,H-6),1.89(2H,m,H-7),1.83(1H,m,H-9),4.66(1H,t,J=10.3Hz,H-11),3.67(1H,d,J=10.3Hz,H-12),2.35(1H,m,H-15a),2.54(1H,m,H-15b),1.89(2H,m,H-16),2.50(1H,m,H-17),1.88(3H,s,Me-18),1.80(3H,s,Me-19),4.09(1H,m,H-20),1.41(3H,d,J=6.4Hz,Me-21),5.28(1H,br d,J=9.3Hz,cym-1),1.86(1H,m,cym-2a),2.27(1H,m,cym-2b),4.00(1H,m,cym-3),3.43(1H,br d,J=8.8Hz,cym-4),4.19(1H,m,cym-5),1.33(3H,d,J=6.4Hz,cym-6),3.55(3H,s,Me-cym-3),5.08(1H,br d,J=9.3Hz,cym-1'),1.80(1H,m,cym-2a'),2.30(1H,m,cym-2b'),4.01(1H,m,cym-3'),3.45(1H,dd,J=8.8Hz,J=2.6Hz,cym-4'),4.15(1H,m,cym-5'),1.43(3H,d,J=6.4Hz,cym-6'),3.58(3H,s,Me-cym-3′),5.07(1H,d,J=8.3Hz,alm-1),3.80(1H,m,alm-2),4.44(1H,m,alm-3),3.71(1H,br d,J=9.8Hz,alm-4),4.24(1H,m,alm-5),1.61(3H,d,J=6.4Hz,cym-6'),3.82(3H,s,Me-alm-3'),4.97(1H,d,J=7.8Hz,glc-1),4.04(1H,m,glc-2),4.23(1H,m,glc-3),4.20(1H,m,glc-4),4.25(1H,m,glc-5),4.37(1H,dd,J=11.7Hz,J=5.4Hz,glc-6)。
13C-NMR(C
5D
5N,125MHz)：δ39.9(t,C-1),30.4(t,C-2),78.3(d,C-3),40.9(t,C-4),141.0(s,C-5),118.6(d,C-6),36.1(t,C-7),76.4(s,C-8),51.1(d,C-9),39.7(s,C-10),70.1(d,C-11),82.2(d,C-12),54.4(s,C-13),85.4(s,C-14),36.0(t,C-15),27.5(t,C-16),56.8(d,C-17),12.2(q，C-18),17.9(q，C-19),69.7(s,C-20),23.3(q,Me-21),96.4(d,cym-1),37.3(t,cym-2),78.1(d,cym-3),83.4(d,cym-4),69.1(d,cym-5),18.7(q,cym-6),58.9(q,Me-cym-3),100.4(d,cym-1'),37.1(t,cym-2'),78.2(d,cym-3'),83.2(d,cym-4'),69.3(d,cym-5'),18.6(q,cym-6'),59.1(q,Me-cym-3'),104.0(d,alm-1),72.5(d,alm-2),83.1(d,alm-3),83.1(d,alm-4),69.3(d,alm-5),18.3(d,alm-6),61.8(q,Me-alm-3'),106.6(d,glc-1),75.5(d,glc-2),78.5(d,glc-3),71.9(d,glc-4),78.4(d,glc-5),63.0(t,glc-6)。
The breeding of zebrafish embryo is carried out in the mode of natural paired cross.Each mating prepares 4 ~ 5 pairs of Adult Zebrafishs, and on average often pair can be produced 200 ~ 300 embryos.At after fertilization 6 hours (i.e. 6hpf) and 24hpf, embryo is cleared up (removing dead embryo), and select suitable embryo according to the etap of embryo.Embryo's (fish culture water water quality: add 200mg Instant Ocean in every 1L reverse osmosis water, specific conductivity is 480 ~ 510 μ S/cm is hatched with fish culture water under 28 DEG C of conditions; PH is 6.9 ~ 7.2; Hardness is 53.7 ~ 71.6mg/LCaCO
3).Because embryo can obtain nutritive substance from the yolk sac of self, so (9dpf) does not need feeding in after fertilization 9 days.After having tested, with tricaine methylsulfonic acid, over-exposure process is carried out to the zebra fish of each etap, thus zebra fish anesthesia is put to death.The operation steps that anesthesia is put to death meets the code requirement that American Veterinary association (AVMA) puts to death Animal Anesthesia.
The zebra fish of Yetrazol process is epilepsy model group; The zebra fish of phenytoin Sodium and Yetrazol co-treatment is positive controls; The zebra fish of the DMSO process of 1.1% is negative control group (solvent control group); Untreated zebra fish is blank group.Root of Twisting Dregea glycosides N is dissolved in DMSO, is mixed with the solution to be measured that concentration is 30 μMs.Above-mentioned solution to be measured respectively with Yetrazol co-treatment zebra fish certain hour, utilize the movement locus of zebra fish in behavioural analysis instrument record 60min, then to the rapid movement (seizure of zebra fish, V>20mm/sec) distance carries out quantitative analysis, by calculating following index: 1. zebra fish rapid movement distance (distance of rapid movement-seizure); 2. the anti-epileptic therapeutic efficiency (therapeutic efficacy) of compound; 3. compound time changing curve (compound effect on the time-course of zebrafish seizure) that zebra fish rapid movement is affected.Root of Twisting Dregea glycosides N and phenytoin Sodium are compared with epilepsy model group, and zebra fish rapid movement track obviously reduces, and tentatively determine that root of Twisting Dregea glycosides N has antiepileptic action, therapeutic efficiency is 19 ± 2.27%.
According to the pathogeny of epilepsy, the zebra fish that the above embodiment of the present invention system selects Yetrazol (Pentylenetetrazole) to process is used as epilepsy model group, testing compound and Yetrazol co-treatment zebra fish certain hour, utilize the motion conditions of zebra fish in motion/behavioural analysis instrument record certain hour, with distance (D), quantitative analysis is carried out to the movement velocity of zebra fish, the anti-epileptic therapeutic efficiency of computerized compound, and draw dose-effect curve according to compound epilepsy therapy efficiency, the IC of computerized compound antiepileptic action
50; Carry out statistical analysis with variance analysis and T-inspection, p<0.05 is significant difference.
Rapid movement refers to that zebra fish movement velocity is greater than the motion of 20mm/sec, is equivalent to mammiferous status epilepticus; Rapid movement track refers to utilize the movement velocity of behavioural analysis instrument record to be greater than the movement locus of 20mm/sec, i.e. the movement locus of zebra fish under status epilepticus, represents with red lines in behavior analytic record result; Rapid movement distance refers to that zebra fish movement velocity is greater than the move distance of 20mm/sec, i.e. the move distance of zebra fish under status epilepticus.
All there are not the zebra fish phenomena of mortality in all experimental group in whole experimentation.
Trial-product selects 11-O-acetyl ester group-12,20-O-diisoamyl acyl ester group-hardship to restrict aglycon and derivative thereof and Cynanchum otophvllum glycoside derivates.
Positive reference substance selects Levetiracetam and Cynanchum otophvllum sheet.
In zebra fish anti-epileptic model, trial-product 11-11-O-acetyl ester group-12,20-O-diisoamyl acyl ester group-hardship restricts aglycon and derivative, Cynanchum otophvllum glycoside derivates and positive control Levetiracetam and Cynanchum otophvllum sheet respectively with DMSO dissolving, and final concentration is 1 μM.Observe each experimental group zebra fish movement locus in 60min, the IC of calculation sample
50.
Example of formulations 1:
By the method first obtained above-claimed cpd of the present invention of embodiment 1-7, and utilize organic acid (tartrate, citric acid, formic acid, oxalic acid etc.) or mineral acid (hydrochloric acid, sulfuric acid, phosphoric acid etc.) salt made, inject with water routinely, essence filter, injection liquid is made in embedding sterilizing.
Example of formulations 2:
By the method first obtained above-claimed cpd of the present invention of embodiment 1-7, and utilize organic acid (tartrate, citric acid, formic acid, oxalic acid etc.) or the salt made of mineral acid (hydrochloric acid, sulfuric acid, phosphoric acid etc.), be dissolved in sterile water for injection, stirring makes molten, filters with aseptic suction funnel, more aseptic essence filter, be sub-packed in 2 ampoules, after frozen drying, aseptic sealing by fusing obtains powder injection.
Example of formulations 3:
By the method first obtained above-claimed cpd of the present invention of embodiment 1-7, and utilize organic acid (tartrate, citric acid, formic acid, oxalic acid etc.) or the salt made of mineral acid (hydrochloric acid, sulfuric acid, phosphoric acid etc.), add vehicle with excipient weight than the ratio for 9:1, make pulvis.
Example of formulations 4:
By the method first obtained above-claimed cpd of the present invention of embodiment 1-7, and utilize organic acid (tartrate, citric acid, formic acid, oxalic acid etc.) or the salt made of mineral acid (hydrochloric acid, sulfuric acid, phosphoric acid etc.), vehicle is added, pelletizing press sheet than the ratio for 1:5-1:10 in itself and excipient weight.
Example of formulations 5:
By the method first obtained above-claimed cpd of the present invention of embodiment 1-7, and utilize organic acid (tartrate, citric acid, formic acid, oxalic acid etc.) or mineral acid (hydrochloric acid, sulfuric acid, phosphoric acid etc.) salt made, oral liquid method for making makes oral liquid routinely.
Example of formulations 6:
By the method first obtained above-claimed cpd of the present invention of embodiment 1-7, and utilize organic acid (tartrate, citric acid, formic acid, oxalic acid etc.) or the salt made of mineral acid (hydrochloric acid, sulfuric acid, phosphoric acid etc.), add vehicle in itself and excipient weight than the ratio for 5:1, make capsule or granule or electuary.
Example of formulations 7:
By the method first obtained above-claimed cpd of the present invention of embodiment 1-7, and utilize organic acid (tartrate, citric acid, formic acid, oxalic acid etc.) or the salt made of mineral acid (hydrochloric acid, sulfuric acid, phosphoric acid etc.), add vehicle in itself and excipient weight than the ratio for 3:1, make capsule or granule or electuary.
Claims (3)
1. the C shown in structural formula (I) ?3,11,12,20 ?four replace C ?21 steroid derivatives and pharmaceutical salts thereof and glycoside,
2. the pharmaceutical composition of antiepileptic, wherein containing C according to claim 1 ?3,11,12,17 ?four replace C ?21 steroid derivative and pharmaceutical salts thereof and glycoside and the pharmaceutically acceptable carrier of at least one.
3. C according to claim 1 ?3,11,12,17 ?four replace C ?21 steroid derivative and pharmaceutical salts thereof and the application of glycoside in the medicine of preparation treatment epileptics.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210419579.6A CN102924554B (en) | 2012-10-29 | 2012-10-29 | C-3,11,12,20-tetrasubstituted C-21 steroid derivative and its pharmaceutical composition and use in medicine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210419579.6A CN102924554B (en) | 2012-10-29 | 2012-10-29 | C-3,11,12,20-tetrasubstituted C-21 steroid derivative and its pharmaceutical composition and use in medicine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102924554A CN102924554A (en) | 2013-02-13 |
| CN102924554B true CN102924554B (en) | 2015-07-22 |
Family
ID=47639504
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210419579.6A Active CN102924554B (en) | 2012-10-29 | 2012-10-29 | C-3,11,12,20-tetrasubstituted C-21 steroid derivative and its pharmaceutical composition and use in medicine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102924554B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105503988B (en) * | 2014-09-22 | 2018-03-30 | 中国科学院上海药物研究所 | Natural antiepileptic activity compound and its purposes in pharmaceutical preparation |
| CN104546877A (en) * | 2014-12-18 | 2015-04-29 | 昆明理工大学 | Application of cynanchum otophyllum scheid saponin M2 in preparation of antiepileptic drugs |
| CN104546878A (en) * | 2014-12-18 | 2015-04-29 | 昆明理工大学 | Medical application of cynanchum otophyllum saponin M1 |
| CN104644663B (en) * | 2014-12-18 | 2017-09-12 | 昆明理工大学 | A kind of cynanchum otophyllum saponin composition and its application |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1148958A (en) * | 1996-08-29 | 1997-05-07 | 中国科学院昆明植物研究所 | Pharmaceutical application of glucoside A and B of auricledleaf swallowort |
| CN1150951A (en) * | 1996-08-29 | 1997-06-04 | 中国科学院昆明植物研究所 | Five compounds of Qingyang ginseng glucoside and its preparation method and application |
| CN1190404A (en) * | 1995-06-06 | 1998-08-12 | 科斯赛斯公司 | Neuroactive steroids of androstane and pregnane series |
| CN101953864A (en) * | 2010-09-29 | 2011-01-26 | 中国人民解放军军事医学科学院毒物药物研究所 | Cynanchum otophyllum aglycone and medical application of extractive containing same |
-
2012
- 2012-10-29 CN CN201210419579.6A patent/CN102924554B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1190404A (en) * | 1995-06-06 | 1998-08-12 | 科斯赛斯公司 | Neuroactive steroids of androstane and pregnane series |
| CN1148958A (en) * | 1996-08-29 | 1997-05-07 | 中国科学院昆明植物研究所 | Pharmaceutical application of glucoside A and B of auricledleaf swallowort |
| CN1150951A (en) * | 1996-08-29 | 1997-06-04 | 中国科学院昆明植物研究所 | Five compounds of Qingyang ginseng glucoside and its preparation method and application |
| CN101953864A (en) * | 2010-09-29 | 2011-01-26 | 中国人民解放军军事医学科学院毒物药物研究所 | Cynanchum otophyllum aglycone and medical application of extractive containing same |
Non-Patent Citations (3)
| Title |
|---|
| "Identification of new qingyangshengenin and caudatin glycosides from the roots of Cynanchum otophyllum";Xiao-Xia Ma et al.;《Steroids》;20110407;第76卷;1003-1009 * |
| "New pregnane glycosides from the roots of Cynanchum otophyllum";Xiao-Xia Ma et al.;《Steroids》;20070704;第72卷;778-786 * |
| "Studies on the Constituents of Asclepiadaceae Plants.LIX.The Structures of Five New Glycosides from Dregea volubilis (L.) BENTH.";Shin-Ichi Yoshimura et al.;《Chem.Pharm.Bull.》;19851231;第33卷(第6期);2287-2293 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102924554A (en) | 2013-02-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| AU2022200876B2 (en) | Enhancing autophagy or increasing longevity by administration of urolithins or precursors thereof | |
| JP5714816B2 (en) | Methods and compositions for inhibiting angiogenesis | |
| CN102924554B (en) | C-3,11,12,20-tetrasubstituted C-21 steroid derivative and its pharmaceutical composition and use in medicine | |
| KR20010030746A (en) | A Steroidal soponins for the prophylaxis or treatment of dementia, and steroidal saponin compounds | |
| PL213644B1 (en) | Use of glycosides of mono- and diacylglycerol as anti-inflammatory agents | |
| CN103933058B (en) | The application of luteolin-7-glucosulfone aldehydic acid glycosides in preparation treatment Retinal degeneration medicine | |
| CN103796647A (en) | Methods and compositions for treating brain cancer | |
| CN103183719B (en) | Saponin derivative and uses thereof | |
| CN105503988B (en) | Natural antiepileptic activity compound and its purposes in pharmaceutical preparation | |
| CN111647003B (en) | Triepoxyhexahydrochromone A, and pharmaceutical composition and application thereof | |
| CN106083972B (en) | A kind of momordica grosvenori alcohol derivatives monomer | |
| CN103524593A (en) | Chikusetsusaponin IVa, derivatives thereof, preparation method thereof and application thereof to preparation of medicines | |
| KR20180119790A (en) | Composition for preventing or treating erectile dysfunction comprising embryonic stem cells derived exosome | |
| CN102266567B (en) | Radix scutellariae extractive phospholipid complex, and preparation method and purpose thereof | |
| CN101665489B (en) | Dehydrosilybin trialky ether and preparation method and medical application thereof | |
| CN106188205B (en) | A kind of purposes of momordica grosvenori alcohol derivatives monomer and combinations thereof | |
| CN111529515B (en) | Application of 12, 15-dioxo-alpha-cnidiene in pharmacy | |
| CN105712982B (en) | Dihydro- β -agarofuran type sesquiterpenoids, preparation method and application thereof | |
| CN108653306B (en) | Application of goldfish grass extract-6-neohesperidoside in treatment of senile dementia | |
| CN103340881B (en) | The application in neuroprotective of a kind of oligosaccharide compound | |
| CN106928299A (en) | One class derives from the compound of the root bark of Chinese wolf-berry, its preparation method and the application in terms of hypoglycemic | |
| CN101565419A (en) | 7 and 20 dehydro-silybin dialky ether and preparation method and medicine use thereof | |
| KR20130075191A (en) | A pharmaceutical composition for inhalation comprising cedrol or the pharmaceutically acceptable salts as an active ingredient for treatment or prevention of cancer | |
| CN111569008B (en) | A pharmaceutical composition for treating neurodegenerative diseases | |
| CN118717747A (en) | Dodelate C lipid lowering application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20130213 Assignee: Beijing Zhongke Wenqing Technology Development Co.,Ltd. Assignor: KUNMING INSTITUTE OF BOTANY, CHINESE ACADEMY OF SCIENCES Contract record no.: X2022530000002 Denomination of invention: C-3,11,12,20-tetrasubstituted C-21 steroid derivatives and their pharmaceutical compositions and their applications in medicine Granted publication date: 20150722 License type: Common License Record date: 20220609 |
|
| EE01 | Entry into force of recordation of patent licensing contract | ||
| OL01 | Intention to license declared | ||
| OL01 | Intention to license declared |