CN105051023B - 促进干细胞分化为肝细胞的化合物 - Google Patents
促进干细胞分化为肝细胞的化合物 Download PDFInfo
- Publication number
- CN105051023B CN105051023B CN201480012330.0A CN201480012330A CN105051023B CN 105051023 B CN105051023 B CN 105051023B CN 201480012330 A CN201480012330 A CN 201480012330A CN 105051023 B CN105051023 B CN 105051023B
- Authority
- CN
- China
- Prior art keywords
- compound
- hydrogen
- hydroxyl
- pyridine
- cell
- 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.)
- Expired - Fee Related
Links
- 150000001875 compounds Chemical class 0.000 title claims abstract description 122
- 210000005229 liver cell Anatomy 0.000 title claims abstract description 41
- 210000000130 stem cell Anatomy 0.000 title claims abstract description 20
- 150000003839 salts Chemical class 0.000 claims abstract description 23
- 239000003814 drug Substances 0.000 claims abstract description 9
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 5
- 241000700721 Hepatitis B virus Species 0.000 claims description 40
- 229910052739 hydrogen Inorganic materials 0.000 claims description 34
- 239000001257 hydrogen Substances 0.000 claims description 34
- 208000015181 infectious disease Diseases 0.000 claims description 24
- 108090000623 proteins and genes Proteins 0.000 claims description 23
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 18
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 15
- 229910052736 halogen Inorganic materials 0.000 claims description 13
- 150000002367 halogens Chemical class 0.000 claims description 13
- 102000014150 Interferons Human genes 0.000 claims description 11
- 108010050904 Interferons Proteins 0.000 claims description 11
- 229940079322 interferon Drugs 0.000 claims description 11
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 10
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 claims description 9
- 229910052731 fluorine Inorganic materials 0.000 claims description 9
- 239000011737 fluorine Substances 0.000 claims description 9
- 150000002431 hydrogen Chemical group 0.000 claims description 6
- 239000000126 substance Substances 0.000 claims description 6
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 5
- 239000000460 chlorine Substances 0.000 claims description 5
- 229910052801 chlorine Inorganic materials 0.000 claims description 5
- SIHUZXVYNKGNFF-HSZRJFAPSA-N N-[(3-fluorophenyl)methyl]-2-[4-[[(2S)-2-hydroxy-2-phenylethyl]amino]-2-oxo-1H-pyridin-3-yl]-1H-benzimidazole-4-carboxamide Chemical compound O[C@H](CNc1cc[nH]c(=O)c1-c1nc2cccc(C(=O)NCc3cccc(F)c3)c2[nH]1)c1ccccc1 SIHUZXVYNKGNFF-HSZRJFAPSA-N 0.000 claims description 3
- 238000004113 cell culture Methods 0.000 claims description 3
- 230000004069 differentiation Effects 0.000 claims description 2
- 238000012216 screening Methods 0.000 claims description 2
- 241000701076 Macacine alphaherpesvirus 1 Species 0.000 claims 4
- 208000002672 hepatitis B Diseases 0.000 claims 4
- 238000000034 method Methods 0.000 abstract description 24
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 12
- 201000010099 disease Diseases 0.000 abstract description 10
- 150000002148 esters Chemical class 0.000 abstract description 7
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 33
- 230000014509 gene expression Effects 0.000 description 21
- 239000000203 mixture Substances 0.000 description 20
- 210000004027 cell Anatomy 0.000 description 19
- BDAGIHXWWSANSR-UHFFFAOYSA-N formic acid Substances OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 19
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 18
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 18
- 239000007787 solid Substances 0.000 description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 15
- WGQKYBSKWIADBV-UHFFFAOYSA-N benzylamine Chemical compound NCC1=CC=CC=C1 WGQKYBSKWIADBV-UHFFFAOYSA-N 0.000 description 14
- 230000015572 biosynthetic process Effects 0.000 description 13
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 13
- 238000003786 synthesis reaction Methods 0.000 description 13
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- 238000005160 1H NMR spectroscopy Methods 0.000 description 10
- 239000001963 growth medium Substances 0.000 description 10
- 238000012545 processing Methods 0.000 description 10
- 239000000376 reactant Substances 0.000 description 10
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 9
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 9
- 241000700605 Viruses Species 0.000 description 9
- 239000002585 base Substances 0.000 description 9
- 230000008859 change Effects 0.000 description 9
- 230000000694 effects Effects 0.000 description 8
- -1 glycolic Chemical compound 0.000 description 8
- RZOFMJXYQYHMAC-MRXNPFEDSA-N 2-[4-[[(2S)-2-hydroxy-2-phenylethyl]amino]-2-oxo-1H-pyridin-3-yl]-1H-benzimidazole-4-carboxylic acid Chemical compound O[C@H](CNc1cc[nH]c(=O)c1-c1nc2cccc(C(O)=O)c2[nH]1)c1ccccc1 RZOFMJXYQYHMAC-MRXNPFEDSA-N 0.000 description 7
- 239000007821 HATU Substances 0.000 description 7
- WMFOQBRAJBCJND-UHFFFAOYSA-M Lithium hydroxide Chemical compound [Li+].[OH-] WMFOQBRAJBCJND-UHFFFAOYSA-M 0.000 description 7
- 230000006870 function Effects 0.000 description 7
- 210000003494 hepatocyte Anatomy 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 6
- SJRJJKPEHAURKC-UHFFFAOYSA-N N-Methylmorpholine Chemical compound CN1CCOCC1 SJRJJKPEHAURKC-UHFFFAOYSA-N 0.000 description 6
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 6
- 239000002253 acid Substances 0.000 description 6
- 238000010586 diagram Methods 0.000 description 6
- 239000000523 sample Substances 0.000 description 6
- QVSVMNXRLWSNGS-UHFFFAOYSA-N (3-fluorophenyl)methanamine Chemical compound NCC1=CC=CC(F)=C1 QVSVMNXRLWSNGS-UHFFFAOYSA-N 0.000 description 5
- 238000010438 heat treatment Methods 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 229910052757 nitrogen Inorganic materials 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- IIFVWLUQBAIPMJ-UHFFFAOYSA-N (4-fluorophenyl)methanamine Chemical compound NCC1=CC=C(F)C=C1 IIFVWLUQBAIPMJ-UHFFFAOYSA-N 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 4
- 125000004429 atom Chemical group 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 125000004175 fluorobenzyl group Chemical group 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 230000003612 virological effect Effects 0.000 description 4
- LRFWYBZWRQWZIM-UHFFFAOYSA-N (2-fluorophenyl)methanamine Chemical compound NCC1=CC=CC=C1F LRFWYBZWRQWZIM-UHFFFAOYSA-N 0.000 description 3
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 3
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- JPYQFYIEOUVJDU-UHFFFAOYSA-N beclamide Chemical compound ClCCC(=O)NCC1=CC=CC=C1 JPYQFYIEOUVJDU-UHFFFAOYSA-N 0.000 description 3
- 235000011089 carbon dioxide Nutrition 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000007876 drug discovery Methods 0.000 description 3
- 239000000706 filtrate Substances 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- 125000001424 substituent group Chemical group 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- WORJRXHJTUTINR-UHFFFAOYSA-N 1,4-dioxane;hydron;chloride Chemical compound Cl.C1COCCO1 WORJRXHJTUTINR-UHFFFAOYSA-N 0.000 description 2
- YZNFGRHAHBWLPH-UHFFFAOYSA-N 2-[4-[2-(3-chlorophenyl)ethylamino]-2-oxo-1H-pyridin-3-yl]-N-[(4-fluorophenyl)methyl]-1H-benzimidazole-4-carboxamide Chemical compound FC1=CC=C(CNC(=O)C2=CC=CC=3N=C(NC32)C=3C(NC=CC3NCCC3=CC(=CC=C3)Cl)=O)C=C1 YZNFGRHAHBWLPH-UHFFFAOYSA-N 0.000 description 2
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 102000008186 Collagen Human genes 0.000 description 2
- 108010035532 Collagen Proteins 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 238000002965 ELISA Methods 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
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 2
- YVXSYNYWDDKDQO-HSZRJFAPSA-N N-benzyl-2-[4-[[(2S)-2-hydroxy-2-phenylethyl]amino]-2-oxo-1H-pyridin-3-yl]-1H-benzimidazole-4-carboxamide Chemical compound O[C@H](CNc1cc[nH]c(=O)c1-c1nc2cccc(C(=O)NCc3ccccc3)c2[nH]1)c1ccccc1 YVXSYNYWDDKDQO-HSZRJFAPSA-N 0.000 description 2
- BHHGXPLMPWCGHP-UHFFFAOYSA-N Phenethylamine Chemical compound NCCC1=CC=CC=C1 BHHGXPLMPWCGHP-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- LCTONWCANYUPML-UHFFFAOYSA-N Pyruvic acid Chemical compound CC(=O)C(O)=O LCTONWCANYUPML-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 150000001412 amines Chemical class 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 229920001436 collagen Polymers 0.000 description 2
- 239000012043 crude product Substances 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 238000010494 dissociation reaction Methods 0.000 description 2
- 230000005593 dissociations Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000001647 drug administration Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 210000003754 fetus Anatomy 0.000 description 2
- 238000003818 flash chromatography Methods 0.000 description 2
- 150000007529 inorganic bases Chemical class 0.000 description 2
- 239000011630 iodine Substances 0.000 description 2
- 229910052740 iodine Inorganic materials 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 201000007270 liver cancer Diseases 0.000 description 2
- 208000019423 liver disease Diseases 0.000 description 2
- 208000014018 liver neoplasm Diseases 0.000 description 2
- QECNBMJRDWINKS-UHFFFAOYSA-N n-fluoro-1-phenylmethanamine Chemical compound FNCC1=CC=CC=C1 QECNBMJRDWINKS-UHFFFAOYSA-N 0.000 description 2
- 150000007530 organic bases Chemical class 0.000 description 2
- 239000002798 polar solvent Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- 159000000000 sodium salts Chemical class 0.000 description 2
- 238000007614 solvation Methods 0.000 description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 2
- GETQZCLCWQTVFV-UHFFFAOYSA-N trimethylamine Chemical compound CN(C)C GETQZCLCWQTVFV-UHFFFAOYSA-N 0.000 description 2
- 230000009385 viral infection Effects 0.000 description 2
- ULSIYEODSMZIPX-MRVPVSSYSA-N (1s)-2-amino-1-phenylethanol Chemical compound NC[C@@H](O)C1=CC=CC=C1 ULSIYEODSMZIPX-MRVPVSSYSA-N 0.000 description 1
- QBYIENPQHBMVBV-HFEGYEGKSA-N (2R)-2-hydroxy-2-phenylacetic acid Chemical compound O[C@@H](C(O)=O)c1ccccc1.O[C@@H](C(O)=O)c1ccccc1 QBYIENPQHBMVBV-HFEGYEGKSA-N 0.000 description 1
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- WBYWAXJHAXSJNI-VOTSOKGWSA-M .beta-Phenylacrylic acid Natural products [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 description 1
- 101150084750 1 gene Proteins 0.000 description 1
- HYZJCKYKOHLVJF-UHFFFAOYSA-N 1H-benzimidazole Chemical compound C1=CC=C2NC=NC2=C1 HYZJCKYKOHLVJF-UHFFFAOYSA-N 0.000 description 1
- RHXSYTACTOMVLJ-UHFFFAOYSA-N 1H-benzimidazole-2-carboxylic acid Chemical compound C1=CC=C2NC(C(=O)O)=NC2=C1 RHXSYTACTOMVLJ-UHFFFAOYSA-N 0.000 description 1
- VVQNAFBGAWCMLU-UHFFFAOYSA-N 1h-benzimidazole-4-carboxylic acid Chemical compound OC(=O)C1=CC=CC2=C1N=CN2 VVQNAFBGAWCMLU-UHFFFAOYSA-N 0.000 description 1
- NRHVNPYOTNGECT-UHFFFAOYSA-N 2-(3-chlorophenyl)ethanamine Chemical class NCCC1=CC=CC(Cl)=C1 NRHVNPYOTNGECT-UHFFFAOYSA-N 0.000 description 1
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 1
- CSDSSGBPEUDDEE-UHFFFAOYSA-N 2-formylpyridine Chemical compound O=CC1=CC=CC=N1 CSDSSGBPEUDDEE-UHFFFAOYSA-N 0.000 description 1
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 description 1
- XTYNIPUFKBBALX-UHFFFAOYSA-N 3-chloro-1h-pyridin-2-one Chemical compound OC1=NC=CC=C1Cl XTYNIPUFKBBALX-UHFFFAOYSA-N 0.000 description 1
- 125000004179 3-chlorophenyl group Chemical group [H]C1=C([H])C(*)=C([H])C(Cl)=C1[H] 0.000 description 1
- HBAQYPYDRFILMT-UHFFFAOYSA-N 8-[3-(1-cyclopropylpyrazol-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl]-3-methyl-3,8-diazabicyclo[3.2.1]octan-2-one Chemical class C1(CC1)N1N=CC(=C1)C1=NNC2=C1N=C(N=C2)N1C2C(N(CC1CC2)C)=O HBAQYPYDRFILMT-UHFFFAOYSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- WBYWAXJHAXSJNI-SREVYHEPSA-N Cinnamic acid Chemical compound OC(=O)\C=C/C1=CC=CC=C1 WBYWAXJHAXSJNI-SREVYHEPSA-N 0.000 description 1
- 206010010356 Congenital anomaly Diseases 0.000 description 1
- 102100039297 Cyclic AMP-responsive element-binding protein 3-like protein 1 Human genes 0.000 description 1
- 241001269238 Data Species 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 1
- 108700039887 Essential Genes Proteins 0.000 description 1
- 101710142246 External core antigen Proteins 0.000 description 1
- 101000745631 Homo sapiens Cyclic AMP-responsive element-binding protein 3-like protein 1 Proteins 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
- PWKSKIMOESPYIA-BYPYZUCNSA-N L-N-acetyl-Cysteine Chemical compound CC(=O)N[C@@H](CS)C(O)=O PWKSKIMOESPYIA-BYPYZUCNSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 1
- DMOCVFXZGGMUPI-HSZRJFAPSA-N N-[(4-fluorophenyl)methyl]-2-[4-[[(2S)-2-hydroxy-2-phenylethyl]amino]-2-oxo-1H-pyridin-3-yl]-1H-benzimidazole-4-carboxamide Chemical compound O[C@H](CNc1cc[nH]c(=O)c1-c1nc2cccc(C(=O)NCc3ccc(F)cc3)c2[nH]1)c1ccccc1 DMOCVFXZGGMUPI-HSZRJFAPSA-N 0.000 description 1
- HTLZVHNRZJPSMI-UHFFFAOYSA-N N-ethylpiperidine Chemical compound CCN1CCCCC1 HTLZVHNRZJPSMI-UHFFFAOYSA-N 0.000 description 1
- UESJVVHIGSVEBJ-UHFFFAOYSA-N O=C(c1cccc2c1nc(C(C(NC=C1)=O)=C1NCCc1cc(Cl)ccc1)[nH]2)NCc1cc(F)ccc1 Chemical compound O=C(c1cccc2c1nc(C(C(NC=C1)=O)=C1NCCc1cc(Cl)ccc1)[nH]2)NCc1cc(F)ccc1 UESJVVHIGSVEBJ-UHFFFAOYSA-N 0.000 description 1
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 1
- IWYDHOAUDWTVEP-UHFFFAOYSA-N R-2-phenyl-2-hydroxyacetic acid Natural products OC(=O)C(O)C1=CC=CC=C1 IWYDHOAUDWTVEP-UHFFFAOYSA-N 0.000 description 1
- 108091006611 SLC10A1 Proteins 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 102100021988 Sodium/bile acid cotransporter Human genes 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 239000003070 absorption delaying agent Substances 0.000 description 1
- 239000000370 acceptor Substances 0.000 description 1
- 229960004308 acetylcysteine Drugs 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- ULSIYEODSMZIPX-UHFFFAOYSA-N alpha-hydroxyphenethylamine Natural products NCC(O)C1=CC=CC=C1 ULSIYEODSMZIPX-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 239000003429 antifungal agent Substances 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 150000001500 aryl chlorides Chemical class 0.000 description 1
- SRSXLGNVWSONIS-UHFFFAOYSA-N benzenesulfonic acid Chemical compound OS(=O)(=O)C1=CC=CC=C1 SRSXLGNVWSONIS-UHFFFAOYSA-N 0.000 description 1
- 229940092714 benzenesulfonic acid Drugs 0.000 description 1
- 229960004365 benzoic acid Drugs 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 1
- 229910052794 bromium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 235000019504 cigarettes Nutrition 0.000 description 1
- 229930016911 cinnamic acid Natural products 0.000 description 1
- 235000013985 cinnamic acid Nutrition 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 229960004106 citric acid Drugs 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 229940125904 compound 1 Drugs 0.000 description 1
- 229940125898 compound 5 Drugs 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- BAPQKUOLQDJQSN-UHFFFAOYSA-N diaminomethyl benzoate Chemical compound NC(N)OC(=O)C1=CC=CC=C1 BAPQKUOLQDJQSN-UHFFFAOYSA-N 0.000 description 1
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 238000012362 drug development process Methods 0.000 description 1
- 238000007877 drug screening Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 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
- 125000004494 ethyl ester group Chemical group 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 235000011087 fumaric acid Nutrition 0.000 description 1
- 229960002598 fumaric acid Drugs 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000005745 host immune response Effects 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 description 1
- JJWLVOIRVHMVIS-UHFFFAOYSA-N isopropylamine Chemical compound CC(C)N JJWLVOIRVHMVIS-UHFFFAOYSA-N 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 229910052744 lithium Inorganic materials 0.000 description 1
- 159000000003 magnesium salts Chemical class 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- 239000011976 maleic acid Substances 0.000 description 1
- 229960002510 mandelic acid Drugs 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 229940098779 methanesulfonic acid Drugs 0.000 description 1
- 238000003808 methanol extraction Methods 0.000 description 1
- 150000004702 methyl esters Chemical class 0.000 description 1
- WBYWAXJHAXSJNI-UHFFFAOYSA-N methyl p-hydroxycinnamate Natural products OC(=O)C=CC1=CC=CC=C1 WBYWAXJHAXSJNI-UHFFFAOYSA-N 0.000 description 1
- 238000004452 microanalysis Methods 0.000 description 1
- 238000002493 microarray Methods 0.000 description 1
- 238000010208 microarray analysis Methods 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000002894 multi-fate stem cell Anatomy 0.000 description 1
- 230000000926 neurological effect Effects 0.000 description 1
- 238000006396 nitration reaction Methods 0.000 description 1
- 238000007339 nucleophilic aromatic substitution reaction Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 239000012044 organic layer Substances 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 150000003053 piperidines Chemical class 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 230000010287 polarization Effects 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 1
- 150000003141 primary amines Chemical class 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 229940107700 pyruvic acid Drugs 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
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 238000011076 safety test Methods 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
- 150000003335 secondary amines Chemical class 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 229960005137 succinic acid Drugs 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 229960001367 tartaric acid Drugs 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 1
- YFTHZRPMJXBUME-UHFFFAOYSA-N tripropylamine Chemical compound CCCN(CCC)CCC YFTHZRPMJXBUME-UHFFFAOYSA-N 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 210000002845 virion Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/067—Hepatocytes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
- A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/20—Antivirals for DNA viruses
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/46—Amines, e.g. putrescine
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2503/00—Use of cells in diagnostics
- C12N2503/02—Drug screening
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2506/00—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
- C12N2506/45—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from artificially induced pluripotent stem cells
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Biochemistry (AREA)
- Cell Biology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Virology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Molecular Biology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Epidemiology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Plural Heterocyclic Compounds (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明涉及式I化合物及其可药用的盐和酯,在式I中,R1–R11如说明书与权利要求书中所定义。此外,本发明还涉及制备和使用式I化合物的方法以及含有此类化合物的药用组合物。式I化合物用于使干细胞分化为更成熟或成人化的肝细胞以用作药物筛选平台和用在疾病模型应用中。
Description
本发明涉及使干细胞分化为更成熟化(adult-like)的肝细胞的化合物、其生产方法以及含有它们的药用组合物。
在药物研发过程中,十分需要体外培养方法(robust in vitro methods)用于肝功能的建模。现有的采用人类原肝细胞培养物的方法具有众所周知的不足之处,即捐赠者与捐赠者间的变化性和功能上的不稳定性。同样地,肝癌细胞系显示了机能上的不足,而且也存在肿瘤细胞系中固有的混杂遗传异常。
尽管多功能干细胞衍生的组织有希望解决捐赠者与捐赠者间变化性的问题,然而迄今为止大多数检测人类诱导性多功能干细胞(hiPSC)衍生性肝细胞的报告都显示,这些细胞相对于成人组织来说与胎儿组织更为相似,这可能会使得将其外推到成人体内的情况更加困难。因此,需要更好的办法使干细胞分化为更成熟或成人化的肝细胞,以生成用于药物发现、药物功效和安全测试的更相关的模型。
使hIPSC成功分化为成人化的肝细胞将促进用于治疗慢性肝病的药物发现工作,此类肝病例如肝炎B病毒(HBV)感染。慢性的HBV(CHB)感染在全世界影响了约3.5亿人口,存在巨大的未被满足的医疗需求。现有的疗法核苷抑制剂和干扰素(IFN)对清除病毒不起作用,并且会产生病毒抗药性和/或不良副作用。基于此类病毒基因组序列的变化性,将HBV分为7种基因型(基因型A–H;A–D为主要基因型)。HBV感染疾病的结果取决于年龄和基因型。因此,大多数CHB感染起因于垂直(母到婴)传递和/或童年感染。与之不同的是,~90%接触病毒的成年人能够在6个月内清除HBV感染。此外,各种临床数据显示病毒基因型影响HBV疾病的进程和对IFN疗法的应答。也同样已知,HBV能够通过各种机制避开宿主免疫反应,此类机制包括下调干扰素刺激基因(ISGs)。对于HBV和宿主先天免疫间复杂的相互作用的更好的理解可能导致产生治疗CHB感染的新的宿主/病毒靶点。然而,探索新的、更加有效的HBV抗病毒药的工作被生理学健全的体外培养系统的缺乏所阻碍。用作生产物(producer)和靶细胞的现有的基于肝癌的系统既不是健全的,也不能包括HBV基因型的多样性。因此,极度需要新的体外系统,这种系统需要在生理学上更相关并且支持(support)所有HBV主要基因型的强烈感染,优选得自临床隔离群。此类系统不仅可以作为药物筛选平台,而且还可以作为HBV疾病模型,包括干扰素响应基因型依赖性的评价。
因此,仍然需要提高干细胞–衍生的肝细胞分化为更成熟的肝细胞,以支持(support)各种基因型导致的患者的HBV强烈感染(robust infection),供作为药物筛选平台和疾病模型使用。
本发明涉及式I化合物及其可药用的盐和酯:
其中R1–R11如下文所定义。此外,本发明涉及制造和使用式I化合物的方法以及含有此类化合物的药用组合物。式I化合物用于使干细胞分化为更成熟或成年化的肝细胞以用作药物筛选平台和在疾病模型平台中使用。
图1提供了热图,显示在采用多剂量实施例1化合物处理后整体肝细胞功能的基因表达增加。生物热图通常用在分子生物学中,以代表对比一些可比样本的许多基因的表达水平(例如不同状态的细胞、不同患者的样本),这些样本获自cDNA样本。图1中,“绿色”表示低表达,而“红色”表示高表达。图示的每行数据是相对的,生成从最低表达水平(绿色)到中等水平(黑色)再到最高表达水平(红色)的梯度。
图2显示,在采用实施例1–7化合物处理后,基于一组成熟相关基因的基因表达,在诱导性多功能干细胞衍生的肝细胞中,各种肝细胞功能的基因表达增加。
图3A和3B显示iCell肝细胞中强烈的HBV感染。图3A为柱状图,显示以实施例1化合物处理的诱导性多功能干细胞衍生的肝细胞引起细胞对HBV感染的敏感性,这种反应以剂量依赖的方式出现。图3B为柱状图,显示病毒感染被干扰素抑制(100IU/ml)。
图4A到4D显示iCell肝细胞中全–基因型的HBV感染,为一系列柱状图,显示以实施例1化合物处理的诱导性多功能干细胞衍生的肝细胞能够支持全部四种HBV主要的基因型(A–D)的强烈感染。维持强烈病毒感染需要实施例1化合物的持续使用。在HBV感染(6d)前采用实施例1化合物预处理细胞6d,或在感染期间预处理6天。干扰素(IFN)用于显示HBV感染的特异性。
图5为柱状图,显示采用实施例1化合物处理的诱导性多功能干细胞衍生的肝细胞支持从患者血清(临床隔离群)中分离出的HBV的感染,而不是细胞培养衍生的病毒(HepG2.2.15)的感染。采用实施例1化合物处理的iCell肝细胞支持患者衍生的HBV感染,而不是细胞培养衍生的HBV感染。
图6A–I到6A–IV涉及HBV的感染性:血清与纯化的病毒,为一系列柱状图,显示移除血清中存在的多余的HBsAg亚病毒颗粒(SVPs)是在采用实施例1化合物处理的诱导性多功能干细胞衍生的肝细胞中获得强烈HBV感染的先决条件。细胞在HBV感染前(6d)采用实施例1化合物预处理6d。
图6B-I和6B–II涉及由过量的HBsAg亚病毒颗粒(SVPs)中纯化HBV病毒颗粒,显示纯化的病毒(戴恩颗粒)由HBsAg SVPs中通过Optiprep梯度超离心分离。采用病毒标记(HBsAg和HBV DNA)和电子显微分析确认病毒的完全纯化。
图7a为采用实施例1化合物处理的诱导性多功能干细胞衍生的肝细胞的微阵列分析(热图)。显示上调和下调>2倍(2hr)、>3倍(24hr)或>6倍(7天)处理后的基因。早在2hr时实施例1化合物下调干扰素–刺激基因(ISGs)。显示在iCell肝细胞对HBV感染易感性中也同样可能起作用的两个基因:显示CREB3L1(早在处理后2hr时引起下调)通过其它病毒(HCV、WNV和DNA病毒)抑制感染细胞的增生,据报道SLC10A1(在处理7天后引起上调)为HBV受体。
图7b涉及实施例1化合物对干扰素–刺激基因(ISGs)的作用,并提供饼图显示实施例1化合物对诱导性多功能干细胞衍生的肝细胞中ISGs表达的动力学效应。975个干扰素–刺激基因(ISGs)的列表以公共数据库中已知的ISGs(参见表1)为基础。
图7c涉及实施例1化合物对ISG表达(975基因)的作用,并提供了饼图,显示采用实施例1化合物处理24hr和7天时所调节的ISGs的例子。975个干扰素–刺激基因(ISGs)的列表以公共数据库中已知的ISGs(参见表1)为基础。
表1显示实施例1化合物对于ISGs在处理2hr、24hr和7天时的动力学效应(p值<0.05)。
表1
除另外说明外,在说明书和权利要求书中的以下具体术语和短语如下定义:
术语“基团”指通过一或多个化学键与另一原子或分子连接从而形成分子中一部分的一个原子或一组化学键合的原子。例如,式I中的变量R1–R11指与式I核心结构通过共价键连接的基团。
当述及具有一或多个氢原子的基团时,术语“取代的”指该基团中至少一个氢原子被另一取代基或基团取代。
术语“任选取代的”指(具有一或多个氢原子的)基团中一或多个氢原子可以但不必须被另一个取代基取代。
术语“卤素”指氟、氯、溴或碘。
除另外说明外,术语“氢”指氢原子(–H)而非H2。
术语“在iCell肝细胞中”指由国际细胞动力学(Cellular DynamicsInternational(CDI))获得的诱导性多功能干细胞衍生肝细胞。
除另外说明外,术语“式化合物”指选自如该式所定义的化合物类的任一化合物(包括任何此类化合物的任一可药用的盐或酯)。
术语“可药用的盐”指保留游离碱或游离酸的生物有效性和性质的那些盐,这些盐不是生物学或其它方面不可取的。此类盐可以由无机酸形成,例如盐酸、氢溴酸、硫酸、硝酸、磷酸等,优选盐酸;由有机酸形成,例如乙酸、丙酸、乙醇酸、丙酮酸、草酸、马来酸、丙二酸、水杨酸、琥珀酸、富马酸、酒石酸、柠檬酸、苯甲酸、肉桂酸、扁桃酸、甲磺酸、乙磺酸、p–甲苯磺酸、N–乙酰基半胱氨酸等。此外,此类盐可以通过向游离酸中加入无机碱或有机碱制备。无机碱衍生的此类盐包括但不限于钠、钾、锂、铵、钙和镁盐等。有机碱衍生的此类盐包括但不限于:伯、仲和叔胺盐、(包括天然存在的取代的胺类)取代的胺类的盐、环胺类的盐和碱性离子交换树脂的盐,例如异丙基胺、三甲基胺、二乙胺、三乙胺、三丙基胺、乙醇胺、赖氨酸、精氨酸、N–乙基哌啶、哌啶、聚胺树脂等的盐。
本发明化合物可以以可药用盐的形式存在。本发明化合物还可以可药用酯(即式I酸的甲酯和乙酯)的形式存在。本发明化合物还可以溶剂化,即水化。溶剂化作用可在生产过程中进行,也可以如由于初始无水式I化合物吸湿性质的结果(水合作用)发生。
具有相同分子式,但其原子键合的性质或顺序不同或其原子的空间排列不同的化合物称为“异构体”。原子空间排列不同的异构体称为“立体异构体”。非对映异构体为在一或多个手性中心具有相反构型且不为对映异构体的立体异构体。具有一或多个互相不可重叠镜像的不对称中心的立体异构体称为“对映异构体”。当化合物具有不对称中心(例如,如果碳原子与四个不同的基团连接)时,可能为一对对映异构体。对映异构体可以以其不对称中心的绝对构型表征,并由Cahn、Ingold和Prelog的R–和S–排序规则或者由分子旋转偏振光平面的方式定义,指定为右旋或左旋(即分别为(+)或(–)–异构体)。手性化合物可以单独对映异构体或其混合物形式存在。含有同等量的对映异构体的混合物被称为“外消旋混合物”。
术语化合物的“治疗有效量”指化合物的量,该量能有效预防、缓解或改善疾病症状,或者延长接受治疗的对象的生存期。治疗有效量的测定在本领域内已知。本发明化合物的治疗有效量或剂量可以在较宽的范围内变化,并且可以以本领域已知的方式决定。此类剂量在每一具体情况中根据患者需求调整,所述具体情况包括服用的具体化合物、给药方式、需要治疗的疾病,以及需要治疗的患者。一般而言,在口服或肠胃外给予体重大约为70Kg的成年人药物时,适当的日剂量为约0.1mg–约5,000mg、1mg–约1,000mg或1mg–100mg,但是当需要时可以超过下限和上限。日剂量可以以单次量或分剂量给药,或者肠胃外给药,也可以连续输注的形式给药。
术语“可药用的载体”可包括任何与药物给药相容的材料,包括溶剂、分散介质、包衣材料、抗细菌和抗真菌剂、等张的和吸收延迟剂以及其它能与药物给药相容的材料和化合物。除了那些与活性化合物不相容的常规介质或成分外,其他任何物质都可以用于本发明组合物中。其他活性化合物也可以加入组合物中。
具体而言,本发明涉及式I化合物及其可药用的盐和酯:
其中R1、R2、R3、R4、R5、R6、R7、R8、R9和R10独立为氢或卤素;且R11为氢或羟基。除另外说明外,式I的化合物包括所有可能的立体异构体(即,(R)–对映异构体、(S)–对映异构体)以及其外消旋和两个对映体的(scalemic)混合物。
在一个实施方案中,R1、R2、R3、R4和R5均为氢。在另一实施方案中,R1、R2、R3、R4或R5中至少一个为卤素。在另一实施方案中,R1、R2、R3、R4或R5中至少一个为氟。在另一实施方案中,R1、R3和R5均为氢,且R2或R4中一个为氟,另一个为氢。
在另一具体实施方案中,R6、R7、R8、R9和R10均为氢。在另一实施方案中,R6、R7、R8、R9和R10中至少一个为卤素。在另一实施方案中,R6、R7、R8、R9和R10中至少一个为氯。在另一实施方案中,R6、R8和R10均为氢,且R7或R9中一个为氯,且另一个为氢。
在一个实施方案中,R11为氢。在一个更具体的实施方案中,R1、R2、R3、R4或R5中一个为卤素(优选氟),其它为氢;且R6、R7、R8、R9、R10和R11为氢。
在另一实施方案中,R11为羟基。在一个更具体的实施方案中,R1、R2、R3、R4或R5中一个为卤素(优选氟),其它为氢;R6、R7、R8、R9和R10为氢,且R11为羟基。
在一个实施方案中,本发明涉及式IA化合物及其可药用的盐和酯:
其中R1、R2、R3、R4、R5、R6、R7、R8、R9和R10独立为氢或卤素;且R11为羟基。
在另一实施方案中,本发明涉及式IB化合物及其可药用的盐和酯:
其中R1、R2、R3、R4、R5、R6、R7、R8、R9和R10独立为氢或卤素;且R11为羟基.
在一个实施方案中,本发明涉及下式化合物:
在另一实施方案中,本发明涉及下式化合物:
在另一实施方案中,本发明涉及下式化合物:
在另一实施方案中,本发明涉及下式化合物:
在另一实施方案中,本发明涉及下式化合物:
在另一实施方案中,本发明涉及下式化合物:
在另一实施方案中,本发明涉及下式化合物:
制备上述化合物的原料和试剂通常可以由例如Aldrich Chemical Co的供应商处获得,或者由本领域技术人员按已知的方法制备。下文中的合成反应流程仅用于说明合成本发明化合物的一些方法,可以对这些合成反应流程进行各种修改,给本领域技术人员提供参考。更多的范例可以参见具体实施例。
本发明化合物可以通过任何常规方法制备。在实施例中提供了合成这些化合物的适当方法。通常而言,式I化合物可以通过下述流程制备。
流程1
由二氨基苯甲酸甲酯2开始(其可由供应商获得或者通过用氢和钯炭还原硝基化合物1制备),使其与吡啶甲醛3缩合,接着就地采用碘进行氧化,得到苯并咪唑4。采用4M盐酸的二氧六环溶液并加热至100℃数小时将苯并咪唑的2–甲氧基–3–碘–吡啶部分转化为3–氯–吡啶酮(pyrmidone)5。可以采用碱如三乙胺或N–甲基吗啉在极性溶剂(如乙腈或N,N–二甲基甲酰胺)中通过亲核芳族取代将化合物5的芳基氯化物取代为2–苯基–乙基胺,并加热数小时。所得化合物可采用例如氢氧化锂在四氢呋喃和水中进行的标准方法去酯化,温和加热得到苯并咪唑羧基酸6。最终化合物如化合物7,可以通过酸6与苄基胺在标准酰胺偶合条件下缩合制备,偶合条件例如N,N–二异丙基–乙基胺和O–(7–氮杂苯并三唑–1–基)–N,N,N’,N–四甲基脲六氟磷酸盐在极性溶剂如二甲基甲酰胺(DMF)中。
实施例
尽管本文描述了某些示例性实施方案,但是本发明化合物也可以采用适当的原料通过本文所述的概括性方法和/或通过本领域普通技术人员已知的方法制备。
实施例1
2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸3–氟–苄基酰胺的合成。
2–(4–碘代–2–甲氧基–吡啶–3–基)–3H–苯并咪唑–4–甲酸甲酯
在250mL圆底烧瓶中,混合2,3–二氨基苯甲酸甲酯(1.5g,9.03mmol)和甲醇(25mL),得到黄色溶液,在氮气环境下搅拌并在水/干冰浴中冷却。向该溶液中逐滴加入4–碘代–2–甲氧基烟醛(2.37g,9.03mmol)的甲醇(15mL)和DMF(10mL)溶液。在滴加过程中,再向反应物中加入更多的甲醇(25.0mL)。在水/干冰浴中放置反应混合物2.5hr,用3hr温热至室温,接着在水/干冰浴中冷却。向该混合物中逐滴加入碘(1.49g,5.87mmol)的甲醇(15mL)溶液,接着将反应物温热至室温过夜。浓缩反应物,采用乙酸乙酯(200mL)和饱和的Na2S2O3(200mL)稀释,并混合。产生明显的不溶物质,过滤混合物。所得的固体采用乙酸乙酯和水洗涤。分离滤液,所得水层采用乙酸乙酯(100mL)和DCM(3×150mL)萃取。有机层经饱和的Na2S2O3和盐水洗涤,合并,并经MgSO4干燥、浓缩为红色油状物/固体。原萃取物中的不溶固体经DCM(5×100mL)洗涤,浓缩滤液,得到深红/黑色固体。将液体提取的粗品和固体提取的粗品溶于最少量的DCM中,混合,并经快速色谱纯化(硅胶,120g,0%–60%乙酸乙酯的己烷溶液洗脱),得到2–(4–碘代–2–甲氧基–吡啶–3–基)–3–H–苯并咪唑–4–甲酸甲酯,为紫色固体,0.73g C15H12IN3O3的LC/MS计算值:(m/e)409.0,实测值:410.0(M+H);1H NMR(DMSO–d6)δ:12.68(s,1H),8.05(d,J=5.5Hz,1H),8.01(d,J=8.0Hz,1H),7.88–7.95(m,1H),7.67(d,J=5.3Hz,1H),7.38(t,J=7.9Hz,1H),3.96(s,3H),3.82(s,3H)。经过DCM萃取后残留的原始不溶固体接着经沸腾的甲醇(5×20ml)萃取。浓缩并干燥甲醇滤液,又得到产物(LCMS测定纯度为83%),为钠盐(假定的)和深紫色固体,0.57g。经过DCM和甲醇萃取后剩余的原始不溶固体又得到产物(LCMS测定纯度为90%),为钠盐(假定的)和深紫色固体,0.88g。合并产率为59%。
2–(4–氯代–2–氧代–1,2–二氢–吡啶–3–基)–3H–苯并咪唑–4–甲酸甲酯
首先进行两个并行的反应,在加热前合并。(在200mL圆底烧瓶中,将2–(4–碘代–2–甲氧基–吡啶–3–基)–3H–苯并咪唑–4–甲酸甲酯(由液体萃取物中分离出的固体)(0.88g,2.15mmol)与1,4–二氧六环(3mL)混合,得到黑色悬浮液。分次加入4M HCl的1,4–二氧六环溶液(14.5mL,58.1mmol),在室温下搅拌混合物17hr。在200mL圆底烧瓶中,将2–(4–碘代–2–甲氧基–吡啶–3–基)–3H–苯并咪唑–4–甲酸甲酯(由快速色谱分离)(0.73g,1.78mmol)与1,4–二氧六环(2mL)混合,得到黑色悬浮液。加入4M HCl的1,4–二氧六环溶液(12mL,48.2mmol),并在室温下搅拌混合物17hr。)将单独的反应混合物与1,4–二氧六环(用于清洗)和4M HCl的1,4–二氧六环溶液(20mL)混合。在油浴中于100℃加热反应混合物3hr,然后冷却至室温。过滤反应物,固体经1,4–二氧六环、水、1,4–二氧六环和己烷洗涤,并经中央真空系统(house vacuum)干燥,得到2–(4–氯代–2–氧代–1,2–二氢–吡啶–3–基)–3H–苯并咪唑–4–甲酸甲酯(0.91g,76.2%产率),为黑色固体。C14H10ClN3O3的LC/MS计算值:(m/e)303.0,实测值:304.1(M+H);1H NMR(DMSO–d6)δ:8.05–8.16(m,2H),8.01(d,J=7.3Hz,1H),7.66–7.76(m,1H),7.50(t,J=7.9Hz,1H),3.92–4.04(m,3H)。
2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸甲酯
在40mL小瓶中,将2–(4–氯代–2–氧代–1,2–二氢–吡啶–3–基)–3H–苯并咪唑–4–甲酸甲酯(0.91g,3.00mmol)、(S)–2–氨基–1–苯基乙醇(822mg,5.99mmol)和N–甲基吗啉(909mg,988,μL,8.99mmol)与DMF(20mL)结合,得到黑色悬浮液。密封小瓶,并在干块(dryblock)中于85℃下加热6.5hr,冷却至室温过周末。反应混合物经水稀释,所得的沉淀物经水和己烷洗涤,得到2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸甲酯(0.87g,71.8%产率),为浅紫色固体。C22H20N4O4的LC/MS计算值:(m/e)404.0,实测值:405.2(M+H);1H NMR(DMSO–d6)δ:13.53(s,1H),11.26(d,J=5.8Hz,1H),10.85(t,J=5.1Hz,1H),7.85(d,J=8.0Hz,1H),7.76–7.82(m,1H),7.55(d,J=7.3Hz,2H),7.34–7.42(m,3H),7.26–7.34(m,2H),6.22(d,J=7.5Hz,1H),5.80(d,J=4.5Hz,1H),4.85–5.00(m,1H),3.98(s,3H),3.64–3.77(m,1H),3.53–3.63(m,1H)。
2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸
在200mL圆底烧瓶中,将2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸甲酯(0.87g,2.15mmol)和LiOH(258mg,10.8mmol)与THF(20mLl)和水(5mL)混合,得到紫色悬浮液。在室温下搅拌反应物过夜。第2天再在干块中于50℃下加热反应混合物3.5hr,并冷却至室温。反应物经水稀释,浓缩,并经更多水稀释,并采用1M HCl酸化,过滤。所得固体经水和己烷洗涤,并经中央真空系统干燥,得到2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸(0.86g,102%产率),为紫色固体。C21H18N4O4的LC/MS计算值:(m/e)390.0,实测值:391.2(M+H);1H NMR;(DMSO–d6)δ:13.35(s,1H),11.19(d,J=6.0Hz,1H),10.97(t,J=4.9Hz,1H),7.75(dd,J=7.7,3.9Hz,2H),7.56(d,J=7.3Hz,2H),7.22–7.44(m,5H),6.20(d,J=7.5Hz,1H),5.80(br.s.,1H),4.92(t,J=5.5Hz,1H),3.54–3.74(m,3H)。
2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸3–氟–苄基酰胺
在100mL圆底烧瓶中,将2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸(0.84g,2.15mmol)、3–氟–苄基胺(296mg,270μL,2.37mmol)和DIEA(612mg,827μL,4.73mmol)和DMF(10mL)混合,得到黑色溶液,并将其加入HATU(982mg,2.58mmol)中。反应混合物在室温下搅拌过夜。第二天,将反应混合物加入水中,并过滤所得沉淀物,经水、乙醚和己烷洗涤。紫色固体不完全溶于最低量的沸腾乙醇中,过滤冷却形成的固体,采用乙醇和己烷洗涤,得到2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸3–氟–苄基酰胺,为浅紫色固体。C28H24FN5O3的LC/MS计算值:(m/e)497.0,实测值:497.9(M+H);1H NMR(DMSO–d6–TFA)δ:11.25(br.s.,1H),10.77(br.s.,1H),9.32(t,J=5.8Hz,1H),7.71–7.97(m,2H),7.14–7.63(m,10H),7.03–7.13(m,1H),6.21(d,J=7.5Hz,1H),4.84(br.s.,1H),4.68(br.s.,2H),3.65(d,J=12.5Hz,1H),3.46(d,J=7.0Hz,1H)。
实施例2
2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸苄基酰胺的合成
采用与2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸3–氟–苄基酰胺相似的方法,由2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸、苄基胺、DIEA、HATU和DMF合成2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸苄基酰胺,得到2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸苄基酰胺。C28H25N5O3的LC/MS计算值:(m/e)479.0,实测值:480(M+H).1HNMR(互变异构体1:2;DMSO–d6)δ:13.38–13.52(m,1H),11.14–11.38(m,1H),10.33–11.02(m,1H),9.18–9.43(m,1H),7.69–7.99(m,2H),7.15–7.61(m,12H),6.12–6.30(m,1H),5.74–5.99(m,1H),4.52–4.96(m,3H),3.49–3.30(m,2H)。
实施例3
2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸4–氟–苄基酰胺的合成
采用与2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸3–氟–苄基酰胺相似的方法,由2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸、苄基胺、DIEA、HATU和DMF合成2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸4–氟–苄基酰胺,得到2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸4–氟–苄基酰胺。C28H24FN5O3的LC/MS计算值:(m/e)497.0,实测值:498(M+H).1H NMR(DMSO–d6)δ:13.35–13.53(m,1H),11.13–11.38(m,1H),10.35–11.03(m,1H),9.19–9.42(m,1H),7.68–7.97(m,2H),7.24–7.58(m,9H),7.08–7.22(m,2H),6.13–6.30(m,1H),5.74–6.02(m,1H),4.49–4.98(m,3H),3.49–3.29(m,2H)。
实施例4
2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸苄基酰胺的合成
采用与2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸甲酯相似的方法,由2–(4–氯代–2–氧代–1,2–二氢–吡啶–3–基)–3H–苯并咪唑–4–甲酸甲酯、2–(3–氯代–苯基)–乙基胺、三乙胺和ACN合成2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸甲酯,得到2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸甲酯。
采用与2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸相似的方法,由2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸甲酯、LiOH、THF和水合成2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸,得到2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸。
采用与2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸3–氟–苄基酰胺相似的方法,由2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸、苄基胺、DIEA、HATU和DMF合成2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸苄基酰胺,得到2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸苄基酰胺。C28H24ClN5O2的LC/MS计算值:(m/e)497.0,实测值:498(M+H)。
实施例5
2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸4–氟–苄基酰胺
采用与2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸3–氟–苄基酰胺相似的方法,由2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–羧羧酸、苄基胺、DIEA、HATU和DMF合成2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸4–氟–苄基酰胺,得到2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸4–氟–苄基酰胺。C28H23ClFN5O2的LC/MS计算值:(m/e)515.0,实测值:516(M+H).1H NMR(互变异构体,DMSO–d6)δ:13.30–13.51(m,1H),11.11–11.49(m,1H),9.98–10.95(m,1H),9.06–9.36(m,1H),7.68–8.00(m,2H),6.93–7.65(m,11H),6.22(d,J=7.3Hz,1H),4.47–4.74(m,2H),3.59–3.85(m,2H),3.05(t,J=6.9Hz,2H)。
实施例6
2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸3–氟–苄基酰胺的合成
采用与2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸3–氟–苄基酰胺相似的方法,由2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸、苄基胺、DIEA、HATU和DMF合成2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸3–氟–苄基酰胺,得到2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–3–羧酸4–氟–苄基酰胺。C28H23ClFN5O2的LC/MS计算值:(m/e)515.0,实测值:516(M+H).1H NMR(互变异构体,DMSO–d6)δ:13.42(s,1H),11.15–11.46(m,1H),10.00–10.91(m,1H),9.08–9.41(m,1H),7.69–8.00(m,2H),6.98–7.59(m,11H),6.22(d,J=7.5Hz,1H),4.49–4.78(m,2H),3.63–3.82(m,2H),3.05(t,J=6.8Hz,2H)。
实施例7
2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸2–氟–苄基酰胺的合成
采用与2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸3–氟–苄基酰胺相似的方法,由2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸、苄基胺、DIEA、HATU和DMF合成2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸2–氟–苄基酰胺,得到2–{4–[2–(3–氯代–苯基)–乙基氨基]–2–氧代–1,2–二氢–吡啶–3–基}–3H–苯并咪唑–4–甲酸2–氟–苄基酰胺。C28H23ClFN5O2的LC/MS计算值:(m/e)515.0,实测值:516(M+H).1H NMR(互变异构体,DMSO–d6)δ:13.33–13.49(m,1H),11.14–11.45(m,1H),10.05–10.92(m,1H),9.08–9.32(m,1H),7.70–7.98(m,2H),7.02–7.61(m,11H),6.16–6.32(m,1H),4.53–4.80(m,2H),3.43–3.84(m,2H),2.75–3.12(m,2H)。
式I化合物具备有价值的特性。已经发现,这些化合物可以用于使干细胞分化为更加成熟或成人化的肝细胞,用于更精确的药物实验和科研。本发明化合物使干细胞分化为更加成熟或成人化肝细胞的活性可以由以下试验证明。此外,还描述了本发明化合物对导致细胞对于HBV的敏感性的宿主基因的作用。
人类诱导多能干细胞的体外实验
使人类iPSC衍生的肝细胞(肝细胞)暴露于式I化合物的作用下,以鉴定能更好模拟成人器官的更大功能性的条件。采用高通量、微流体定量RT–PCR(qRT–PCR)测定贯穿肝细胞功能谱的32种基因的表达,其相较于成年的原代人类肝细胞,在hiPSC衍生的肝细胞中是低表达的或者显示不成熟的表型。在初级筛查中,鉴定出许多化合物能够引起许多成熟相关基因的显著增加。在二次筛查中,验证并确认了基因表达的变化,且查询了功能性结果。
细胞和培养条件
根据iCell肝细胞解离和接入用户指南,将新鲜的肝细胞(第20–23天)以每孔60k个细胞接种到96孔BIO胶原IV包被的培养板(BD Cat#354429)中,并进行培养。接种4小时后,移出培养基C,替换为在培养基D中的1:50Matrigel(Cat#354227)覆盖。接种24小时后,向在培养基D和1%DMSO中的5uM化合物施用至细胞。第三天,移出培养基并再加入5uM化合物。第四天,收获RNA。
基因表达谱
采用Gene Expression Cells–to–CTTM Kit(Life TechnologiesCat#4387299)分离样本RNA,在经化合物处理后的多个时间点于–80℃冷冻。所有样品都采用Biomark Fluidigm 96.96芯片(BMK–M–96.96)和ABI Taqman探针通过微流体定量PCR处理。使用Biogazelle qBASE和Genorm软件计算归一化和基于模型的表达测量值。所有样品数据均为一式三份的平均数并且相对于5个管家基因归一化以获得相对基因表达值。用相对于溶媒对照的倍数变化来计算表达值。参见图1和2。
基于化合物改变基因表达的能力选择高击中的化合物(top compound hits),预计化合物通过增加细胞成熟度改变表达,例如增加成年特异标志物,或者降低胎儿特异标志物。在二次确认筛查中,通过更广泛基因的剂量反应选择化合物。结果发现,实施例1化合物在多个剂量时引起跨肝细胞功能的基因的总体增加(图1)。基于一组成熟相关基因的基因表达,暴露于实施例1化合物和其它五个结构类似物(实施例2–7)引起iCell肝细胞的相似表型改变(图2)。采用实施例1化合物的结果显示,在5+独立批次的iCell肝细胞上可再现的基因表达改变,并对此进行了进一步研究,以确定其作用机制和功能性结果。经实施例1化合物处理,根据IHV和ELISA,iCell肝细胞能被多种HBV基因型感染,并形成大量的感染的肝细胞。
微阵列分析
经实施例1化合物处理的iCell肝细胞引起大量基因的上调和下调;包括干扰素–刺激基因(ISGs)表达的动力学效应。参见图7a、7b、7c和表1。
血清中HBV的纯化
将200微升含有HBV的血清放入10–50%Optiprep梯度的SW41管中。以100,000×g于4℃离心样品2hr。在顶部收集500微升级分,对每一级分分析HBsAg(ELISA)和HBV DNA(TaqMan PCR)。含有病毒的级分在–80℃储存。参见图6b。
用HBV对iCell肝细胞进行感染
根据iCell肝细胞解离和接入用户指南,将新鲜的肝细胞(第20–23天)以每孔60k个细胞接种到96孔BIO胶原IV包被的培养板(BD Cat#354429)中,并进行培养。接种4小时后,移出培养基C,替换为在培养基D中的1:50Matrigel(Cat#354227)覆盖。接种24小时后,用在含有1%DMSO的培养基D中的1uM实施例1化合物处理细胞。2天后补充含有新鲜化合物的培养基。接种4天后,采用MOI(感染复数)为10的HBV对细胞进行感染。简而言之,将纯化的病毒在含有实施例1化合物的培养基D中稀释,并与细胞一起温育4–6hr或过夜。移出病毒接种物后,加入含有1uM实施例1化合物的新鲜培养基,温育细胞14天,每两天进行一次培养基更换。分析培养基中分泌的病毒抗原(HBsAg,HBeAg)和HBV DNA。参见图3、4、5、6a。
总之,数据显示,采用式I化合物作为内源的信号提供了快速、有效、非遗传且成本划算的方法以调节iCell肝细胞功能。采用式I化合物产生感染HBV的iCell肝细胞为基础病毒学和药物发现提供了方法。用于干细胞衍生的细胞的功能改进的小分子库筛选可导致产生用于药物发现的新一代体外试验测试方法。
Claims (30)
1.式I化合物或其可药用的盐:
其中R1、R2、R3、R4、R5、R6、R7、R8、R9和R10独立为氢或卤素;且R11为氢或羟基。
2.权利要求1的化合物,其中R1、R2、R3、R4和R5均为氢。
3.权利要求1的化合物,其中R1、R2、R3、R4或R5中至少一个为卤素。
4.权利要求1的化合物,其中R1、R2、R3、R4或R5中至少一个为氟。
5.权利要求1的化合物,其中R1、R3和R5均为氢,且R2或R4中一个为氟,另一个为氢。
6.权利要求1的化合物,其中R6、R7、R8、R9和R10均为氢。
7.权利要求1的化合物,其中R6、R7、R8、R9和R10中至少一个为卤素。
8.权利要求1的化合物,其中R6、R7、R8、R9和R10中至少一个为氯。
9.权利要求1的化合物,其中R6、R8和R10均为氢,且R7或R9中一个为氯,另一个为氢。
10.权利要求1的化合物,其中R11为氢。
11.权利要求1的化合物,其中R1、R2、R3、R4或R5中的一个为氟,其它为氢;且R6、R7、R8、R9、R10和R11为氢。
12.权利要求1的化合物,其中R11为羟基。
13.权利要求1的化合物,其中R1、R2、R3、R4或R5为氟,其它为 氢;R6、R7、R8、R9和R10为氢,且R11为羟基。
14.权利要求1的化合物,其为下式的化合物:
15.权利要求1的化合物,其为下式的化合物:
16.权利要求1的化合物,其为下式的化合物:
17.权利要求1的化合物,其为下式的化合物:
18.权利要求1的化合物,其为下式的化合物:
19.权利要求1的化合物,其为下式的化合物:
20.权利要求1的化合物,其为下式的化合物:
21.权利要求1的化合物,其为式IA的化合物及其可药用的盐:
其中R1、R2、R3、R4、R5、R6、R7、R8、R9和R10独立为氢或卤素;且R11为氢或羟基。
22.权利要求1的化合物,其为式IB的化合物及其可药用的盐:
其中R1、R2、R3、R4、R5、R6、R7、R8、R9和R10独立为氢或卤素;且R11为氢或羟基。
23.药用组合物,该药用组合物包含权利要求1的化合物及可药用的载体。
24.权利要求1的化合物在制备使干细胞分化为肝细胞的药物中的用途。
25.权利要求24中的用途,其中所述肝细胞被乙型肝炎病毒感染。
26.权利要求25中的用途,其中所述感染的肝细胞用于筛选治疗乙型肝炎病毒的化合物。
27.权利要求24中的用途,其中干扰素–刺激基因在分化的肝细胞中 被下调。
28.权利要求27中的用途,其中所述肝细胞被乙型肝炎病毒感染。
29.权利要求25中的用途,其中所述乙型肝炎病毒为患者衍生的乙型肝炎病毒,而不是细胞培养衍生的乙型肝炎病毒。
30.权利要求24中的用途,其中所述化合物为2–[4–((S)–2–羟基–2–苯基–乙基氨基)–2–氧代–1,2–二氢–吡啶–3–基]–3H–苯并咪唑–4–甲酸3–氟–苄基酰胺。
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201361792019P | 2013-03-15 | 2013-03-15 | |
US61/792,019 | 2013-03-15 | ||
US201361811155P | 2013-05-22 | 2013-05-22 | |
US61/811,155 | 2013-05-22 | ||
PCT/EP2014/054763 WO2014140058A1 (en) | 2013-03-15 | 2014-03-12 | Compounds for improved stem cell differentiation into hepatocytes |
Publications (2)
Publication Number | Publication Date |
---|---|
CN105051023A CN105051023A (zh) | 2015-11-11 |
CN105051023B true CN105051023B (zh) | 2017-08-22 |
Family
ID=50272618
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201480012330.0A Expired - Fee Related CN105051023B (zh) | 2013-03-15 | 2014-03-12 | 促进干细胞分化为肝细胞的化合物 |
Country Status (11)
Country | Link |
---|---|
US (2) | US9334480B2 (zh) |
EP (1) | EP2970178B1 (zh) |
JP (1) | JP6129360B2 (zh) |
KR (1) | KR101767802B1 (zh) |
CN (1) | CN105051023B (zh) |
BR (1) | BR112015023185A2 (zh) |
CA (1) | CA2900690A1 (zh) |
HK (1) | HK1217199A1 (zh) |
MX (1) | MX346497B (zh) |
RU (1) | RU2662954C2 (zh) |
WO (1) | WO2014140058A1 (zh) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP3720441A1 (en) * | 2017-12-04 | 2020-10-14 | H. Hoffnabb-La Roche Ag | Pyrrolo[2,3-b]pyrazine compounds as cccdna inhibitors for the treatment of hepatitis b virus (hbv) infection |
JP2023516484A (ja) | 2020-03-11 | 2023-04-19 | ビット バイオ リミテッド | 肝細胞作製方法 |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7498171B2 (en) * | 2002-04-12 | 2009-03-03 | Anthrogenesis Corporation | Modulation of stem and progenitor cell differentiation, assays, and uses thereof |
WO2005024004A1 (ja) * | 2003-08-27 | 2005-03-17 | Renomedix Institute Inc. | 間葉系幹細胞の肝細胞への分化方法及び人工ヒト肝臓細胞 |
EP2036889A4 (en) * | 2006-05-23 | 2009-11-04 | Alla Chem Llc | SUBSTITUTED INDOLES AND PROCESS FOR PRODUCTION AND USE THEREOF |
JP6008297B2 (ja) * | 2011-04-15 | 2016-10-19 | 国立大学法人鳥取大学 | ヒト間葉系幹細胞を肝細胞へ分化誘導する新規化合物の合成と解析 |
-
2014
- 2014-03-12 EP EP14709646.5A patent/EP2970178B1/en not_active Not-in-force
- 2014-03-12 MX MX2015011505A patent/MX346497B/es active IP Right Grant
- 2014-03-12 KR KR1020157025369A patent/KR101767802B1/ko active IP Right Grant
- 2014-03-12 JP JP2015562101A patent/JP6129360B2/ja not_active Expired - Fee Related
- 2014-03-12 WO PCT/EP2014/054763 patent/WO2014140058A1/en active Application Filing
- 2014-03-12 BR BR112015023185A patent/BR112015023185A2/pt not_active IP Right Cessation
- 2014-03-12 RU RU2015140469A patent/RU2662954C2/ru not_active IP Right Cessation
- 2014-03-12 US US14/205,893 patent/US9334480B2/en not_active Expired - Fee Related
- 2014-03-12 CA CA2900690A patent/CA2900690A1/en not_active Abandoned
- 2014-03-12 CN CN201480012330.0A patent/CN105051023B/zh not_active Expired - Fee Related
-
2015
- 2015-02-03 US US14/612,893 patent/US9181218B2/en not_active Expired - Fee Related
-
2016
- 2016-05-09 HK HK16105247.2A patent/HK1217199A1/zh not_active IP Right Cessation
Non-Patent Citations (2)
Title |
---|
Identification of 1-isopropylsulfonyl-2-amine benzimidazoles as a new class of inhibitors of hepatitis B virus;Yun-Fei Li,et al.,;《European Journal of Medicinal Chemistry》;20070331;第42卷;第1358-1364页 * |
Synthesis and Anti-Hepatitis B Virus Activity of Novel Benzimidazole Derivatives;Yun-Fei Li,et al.,;《J. Med. Chem.》;20060627;第49卷;第4790-4794页 * |
Also Published As
Publication number | Publication date |
---|---|
KR20150119342A (ko) | 2015-10-23 |
RU2662954C2 (ru) | 2018-07-31 |
JP6129360B2 (ja) | 2017-05-17 |
US9181218B2 (en) | 2015-11-10 |
CA2900690A1 (en) | 2014-09-18 |
US9334480B2 (en) | 2016-05-10 |
BR112015023185A2 (pt) | 2017-07-18 |
MX346497B (es) | 2017-03-22 |
WO2014140058A1 (en) | 2014-09-18 |
KR101767802B1 (ko) | 2017-08-11 |
RU2015140469A (ru) | 2017-04-21 |
MX2015011505A (es) | 2016-01-22 |
HK1217199A1 (zh) | 2016-12-30 |
CN105051023A (zh) | 2015-11-11 |
JP2016511276A (ja) | 2016-04-14 |
US20150158840A1 (en) | 2015-06-11 |
EP2970178A1 (en) | 2016-01-20 |
EP2970178B1 (en) | 2017-06-14 |
US20150197726A1 (en) | 2015-07-16 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
WO2018024208A1 (zh) | Ido1抑制剂及其制备方法和应用 | |
CN105051023B (zh) | 促进干细胞分化为肝细胞的化合物 | |
CN114174271B (zh) | 吡嗪-2(1h)-酮类化合物的c晶型和e晶型及其制备方法 | |
CN104804079B (zh) | 伊马替尼免疫原、衍生物及合成方法、特异性抗体和检测试剂及制备方法 | |
WO2022148243A1 (zh) | 一种嘧啶类小分子化合物及其应用 | |
CN113200956B (zh) | 一种磺胺苯甲酰胺类衍生物及其制备方法和应用 | |
CN104910894B (zh) | 一种苯并咪唑类hERG钾离子通道的小分子荧光探针及其制备方法与应用 | |
CN101585799B (zh) | 一种制备不对称双吲哚甲烷类化合物的方法 | |
CN111518104B (zh) | 一种含硫脲嘧啶的1,2,4-三氮唑并[1,5-a]嘧啶类化合物及其制备方法和应用 | |
CN107841532B (zh) | 筛选抗乳腺癌转移化合物的方法及相关化合物的应用 | |
CN105837596A (zh) | 双重hdac/brd4抑制剂及其制备方法和应用 | |
CN103319403B (zh) | 一种肿瘤血管新生抑制剂己啉酮及其制备方法和用途 | |
CN107163028A (zh) | 一种苯甲酰胺类Hedgehog抑制剂及其制备方法和应用 | |
CN110204539A (zh) | 一种二氢嘧啶类前药及其制备方法和应用 | |
CN109305961A (zh) | 具有药物活性的伊马胺衍生物及其制备方法 | |
CN108727400A (zh) | 一种治疗肿瘤的化合物 | |
US11964969B2 (en) | Crystal forms of thiazole compound and application thereof | |
CN108659077A (zh) | 核苷磷酰胺类化合物的晶型及制法、用途、衍生组合物 | |
US20220402939A1 (en) | Crystal form of hepatitis b surface antigen inhibitor and application thereof | |
CN111875558B (zh) | 一种噻唑胺衍生物及其抗抑郁的用途 | |
CN111925398B (zh) | 一种fto小分子抑制剂钯配合物及其合成方法 | |
WO2022078307A1 (zh) | 多取代苯环化合物马来酸盐的晶型、其制备方法及其应用 | |
WO2023083357A1 (zh) | 氮杂稠环酰胺类化合物的盐、其结晶形式及其用途 | |
CN105585527B (zh) | 2-(3,4-二氢异喹啉-1(2h)-亚基)乙腈类化合物及其应用 | |
CN110483493A (zh) | 一种二唑类衍生物及其制备方法和应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
REG | Reference to a national code |
Ref country code: HK Ref legal event code: DE Ref document number: 1217199 Country of ref document: HK |
|
GR01 | Patent grant | ||
GR01 | Patent grant | ||
REG | Reference to a national code |
Ref country code: HK Ref legal event code: GR Ref document number: 1217199 Country of ref document: HK |
|
CF01 | Termination of patent right due to non-payment of annual fee | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170822 |