CN117337286A - Methylsulfonyl urea compound and application thereof - Google Patents
Methylsulfonyl urea compound and application thereof Download PDFInfo
- Publication number
- CN117337286A CN117337286A CN202280033410.9A CN202280033410A CN117337286A CN 117337286 A CN117337286 A CN 117337286A CN 202280033410 A CN202280033410 A CN 202280033410A CN 117337286 A CN117337286 A CN 117337286A
- Authority
- CN
- China
- Prior art keywords
- compound
- mol
- added
- water
- stirred
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- -1 Methylsulfonyl urea compound Chemical class 0.000 title abstract description 14
- 150000001875 compounds Chemical class 0.000 claims abstract description 377
- 150000003839 salts Chemical class 0.000 claims abstract description 36
- 239000003814 drug Substances 0.000 claims description 20
- 125000000217 alkyl group Chemical group 0.000 claims description 14
- 125000003545 alkoxy group Chemical group 0.000 claims description 12
- 208000002672 hepatitis B Diseases 0.000 claims description 12
- 229910052736 halogen Inorganic materials 0.000 claims description 9
- 150000002367 halogens Chemical class 0.000 claims description 9
- 238000006467 substitution reaction Methods 0.000 claims description 9
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 8
- 229910052731 fluorine Inorganic materials 0.000 claims description 7
- 125000000113 cyclohexyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C1([H])[H] 0.000 claims description 6
- 125000006546 (C4-C10) cycloalkyl group Chemical group 0.000 claims description 5
- 125000001995 cyclobutyl group Chemical group [H]C1([H])C([H])([H])C([H])(*)C1([H])[H] 0.000 claims description 5
- 201000010099 disease Diseases 0.000 claims description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 5
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 4
- 125000001570 methylene group Chemical group [H]C([H])([*:1])[*:2] 0.000 claims description 3
- 208000000419 Chronic Hepatitis B Diseases 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 315
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 151
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 137
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 128
- 238000006243 chemical reaction Methods 0.000 description 126
- 239000000243 solution Substances 0.000 description 111
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 102
- 230000015572 biosynthetic process Effects 0.000 description 101
- 238000003786 synthesis reaction Methods 0.000 description 101
- 230000002829 reductive effect Effects 0.000 description 97
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 86
- 239000011541 reaction mixture Substances 0.000 description 79
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 78
- 238000005481 NMR spectroscopy Methods 0.000 description 74
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 70
- 239000012043 crude product Substances 0.000 description 70
- 239000012074 organic phase Substances 0.000 description 64
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 62
- 239000000203 mixture Substances 0.000 description 55
- 239000012071 phase Substances 0.000 description 54
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 52
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 52
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 51
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 49
- 239000003208 petroleum Substances 0.000 description 43
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 42
- 238000000605 extraction Methods 0.000 description 40
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 39
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 38
- 229910052757 nitrogen Inorganic materials 0.000 description 38
- 210000004027 cell Anatomy 0.000 description 37
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 32
- 239000000706 filtrate Substances 0.000 description 32
- 238000002953 preparative HPLC Methods 0.000 description 32
- DLYUQMMRRRQYAE-UHFFFAOYSA-N tetraphosphorus decaoxide Chemical compound O1P(O2)(=O)OP3(=O)OP1(=O)OP2(=O)O3 DLYUQMMRRRQYAE-UHFFFAOYSA-N 0.000 description 32
- 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 31
- 235000019253 formic acid Nutrition 0.000 description 31
- 238000003818 flash chromatography Methods 0.000 description 28
- 230000014759 maintenance of location Effects 0.000 description 22
- 210000003813 thumb Anatomy 0.000 description 17
- 238000004440 column chromatography Methods 0.000 description 16
- 230000003833 cell viability Effects 0.000 description 15
- 239000000523 sample Substances 0.000 description 15
- MZRVEZGGRBJDDB-UHFFFAOYSA-N N-Butyllithium Chemical compound [Li]CCCC MZRVEZGGRBJDDB-UHFFFAOYSA-N 0.000 description 14
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 13
- 238000004458 analytical method Methods 0.000 description 13
- 229940079593 drug Drugs 0.000 description 13
- 238000012360 testing method Methods 0.000 description 13
- 241000699670 Mus sp. Species 0.000 description 12
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 description 12
- 239000003480 eluent Substances 0.000 description 12
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical group CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 10
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 10
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
- 238000002474 experimental method Methods 0.000 description 10
- 125000001424 substituent group Chemical group 0.000 description 10
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical class [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 9
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 9
- 239000008280 blood Substances 0.000 description 9
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 8
- 239000002253 acid Substances 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- 229910052739 hydrogen Inorganic materials 0.000 description 8
- 239000001257 hydrogen Substances 0.000 description 8
- KDLHZDBZIXYQEI-UHFFFAOYSA-N palladium Substances [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 8
- XHXFXVLFKHQFAL-UHFFFAOYSA-N phosphoryl trichloride Chemical compound ClP(Cl)(Cl)=O XHXFXVLFKHQFAL-UHFFFAOYSA-N 0.000 description 8
- 238000003756 stirring Methods 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 7
- SHBUUTHKGIVMJT-UHFFFAOYSA-N Hydroxystearate Chemical compound CCCCCCCCCCCCCCCCCC(=O)OO SHBUUTHKGIVMJT-UHFFFAOYSA-N 0.000 description 7
- 239000004698 Polyethylene Substances 0.000 description 7
- 229910052799 carbon Inorganic materials 0.000 description 7
- 239000012065 filter cake Substances 0.000 description 7
- 229940072106 hydroxystearate Drugs 0.000 description 7
- 229920000573 polyethylene Polymers 0.000 description 7
- 229920006395 saturated elastomer Polymers 0.000 description 7
- QFLWZFQWSBQYPS-AWRAUJHKSA-N (3S)-3-[[(2S)-2-[[(2S)-2-[5-[(3aS,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]-3-methylbutanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-4-[1-bis(4-chlorophenoxy)phosphorylbutylamino]-4-oxobutanoic acid Chemical compound CCCC(NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)[C@@H](NC(=O)CCCCC1SC[C@@H]2NC(=O)N[C@H]12)C(C)C)P(=O)(Oc1ccc(Cl)cc1)Oc1ccc(Cl)cc1 QFLWZFQWSBQYPS-AWRAUJHKSA-N 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 6
- 229910002091 carbon monoxide Inorganic materials 0.000 description 6
- 230000005764 inhibitory process Effects 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- SCVFZCLFOSHCOH-UHFFFAOYSA-M potassium acetate Chemical compound [K+].CC([O-])=O SCVFZCLFOSHCOH-UHFFFAOYSA-M 0.000 description 6
- 238000005160 1H NMR spectroscopy Methods 0.000 description 5
- RKMGAJGJIURJSJ-UHFFFAOYSA-N 2,2,6,6-Tetramethylpiperidine Substances CC1(C)CCCC(C)(C)N1 RKMGAJGJIURJSJ-UHFFFAOYSA-N 0.000 description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 5
- 238000011529 RT qPCR Methods 0.000 description 5
- 125000004122 cyclic group Chemical group 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 5
- 125000005647 linker group Chemical group 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- PIBWKRNGBLPSSY-UHFFFAOYSA-L palladium(II) chloride Chemical compound Cl[Pd]Cl PIBWKRNGBLPSSY-UHFFFAOYSA-L 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 5
- 238000000926 separation method Methods 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- KZPYGQFFRCFCPP-UHFFFAOYSA-N 1,1'-bis(diphenylphosphino)ferrocene Chemical compound [Fe+2].C1=CC=C[C-]1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=C[C-]1P(C=1C=CC=CC=1)C1=CC=CC=C1 KZPYGQFFRCFCPP-UHFFFAOYSA-N 0.000 description 4
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 125000004432 carbon atom Chemical group C* 0.000 description 4
- 231100000135 cytotoxicity Toxicity 0.000 description 4
- 230000003013 cytotoxicity Effects 0.000 description 4
- YNHIGQDRGKUECZ-UHFFFAOYSA-N dichloropalladium;triphenylphosphanium Chemical compound Cl[Pd]Cl.C1=CC=CC=C1[PH+](C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1[PH+](C=1C=CC=CC=1)C1=CC=CC=C1 YNHIGQDRGKUECZ-UHFFFAOYSA-N 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- GPAYUJZHTULNBE-UHFFFAOYSA-N diphenylphosphine Chemical compound C=1C=CC=CC=1PC1=CC=CC=C1 GPAYUJZHTULNBE-UHFFFAOYSA-N 0.000 description 4
- 238000010828 elution Methods 0.000 description 4
- KTWOOEGAPBSYNW-UHFFFAOYSA-N ferrocene Chemical compound [Fe+2].C=1C=C[CH-]C=1.C=1C=C[CH-]C=1 KTWOOEGAPBSYNW-UHFFFAOYSA-N 0.000 description 4
- 125000000524 functional group Chemical group 0.000 description 4
- 210000004185 liver Anatomy 0.000 description 4
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 4
- 231100000252 nontoxic Toxicity 0.000 description 4
- 230000003000 nontoxic effect Effects 0.000 description 4
- NFHFRUOZVGFOOS-UHFFFAOYSA-N palladium;triphenylphosphane Chemical compound [Pd].C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 NFHFRUOZVGFOOS-UHFFFAOYSA-N 0.000 description 4
- 239000008363 phosphate buffer Substances 0.000 description 4
- 229910000029 sodium carbonate Inorganic materials 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 230000001988 toxicity Effects 0.000 description 4
- 231100000419 toxicity Toxicity 0.000 description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 4
- ASGMFNBUXDJWJJ-JLCFBVMHSA-N (1R,3R)-3-[[3-bromo-1-[4-(5-methyl-1,3,4-thiadiazol-2-yl)phenyl]pyrazolo[3,4-d]pyrimidin-6-yl]amino]-N,1-dimethylcyclopentane-1-carboxamide Chemical compound BrC1=NN(C2=NC(=NC=C21)N[C@H]1C[C@@](CC1)(C(=O)NC)C)C1=CC=C(C=C1)C=1SC(=NN=1)C ASGMFNBUXDJWJJ-JLCFBVMHSA-N 0.000 description 3
- STBLNCCBQMHSRC-BATDWUPUSA-N (2s)-n-[(3s,4s)-5-acetyl-7-cyano-4-methyl-1-[(2-methylnaphthalen-1-yl)methyl]-2-oxo-3,4-dihydro-1,5-benzodiazepin-3-yl]-2-(methylamino)propanamide Chemical compound O=C1[C@@H](NC(=O)[C@H](C)NC)[C@H](C)N(C(C)=O)C2=CC(C#N)=CC=C2N1CC1=C(C)C=CC2=CC=CC=C12 STBLNCCBQMHSRC-BATDWUPUSA-N 0.000 description 3
- UDQTXCHQKHIQMH-KYGLGHNPSA-N (3ar,5s,6s,7r,7ar)-5-(difluoromethyl)-2-(ethylamino)-5,6,7,7a-tetrahydro-3ah-pyrano[3,2-d][1,3]thiazole-6,7-diol Chemical compound S1C(NCC)=N[C@H]2[C@@H]1O[C@H](C(F)F)[C@@H](O)[C@@H]2O UDQTXCHQKHIQMH-KYGLGHNPSA-N 0.000 description 3
- UIADWSXPNFQQCZ-UHFFFAOYSA-N 4-[4-(3,4-dichlorophenyl)-5-phenyl-1,3-oxazol-2-yl]butanoic acid Chemical compound ClC=1C=C(C=CC=1Cl)C=1N=C(OC=1C1=CC=CC=C1)CCCC(=O)O UIADWSXPNFQQCZ-UHFFFAOYSA-N 0.000 description 3
- 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 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- BQXUPNKLZNSUMC-YUQWMIPFSA-N CCN(CCCCCOCC(=O)N[C@H](C(=O)N1C[C@H](O)C[C@H]1C(=O)N[C@@H](C)c1ccc(cc1)-c1scnc1C)C(C)(C)C)CCOc1ccc(cc1)C(=O)c1c(sc2cc(O)ccc12)-c1ccc(O)cc1 Chemical compound CCN(CCCCCOCC(=O)N[C@H](C(=O)N1C[C@H](O)C[C@H]1C(=O)N[C@@H](C)c1ccc(cc1)-c1scnc1C)C(C)(C)C)CCOc1ccc(cc1)C(=O)c1c(sc2cc(O)ccc12)-c1ccc(O)cc1 BQXUPNKLZNSUMC-YUQWMIPFSA-N 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 3
- 229940127007 Compound 39 Drugs 0.000 description 3
- YZCKVEUIGOORGS-OUBTZVSYSA-N Deuterium Chemical compound [2H] YZCKVEUIGOORGS-OUBTZVSYSA-N 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- 102000006833 Multifunctional Enzymes Human genes 0.000 description 3
- 108010047290 Multifunctional Enzymes Proteins 0.000 description 3
- 108010019160 Pancreatin Proteins 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- LJOOWESTVASNOG-UFJKPHDISA-N [(1s,3r,4ar,7s,8s,8as)-3-hydroxy-8-[2-[(4r)-4-hydroxy-6-oxooxan-2-yl]ethyl]-7-methyl-1,2,3,4,4a,7,8,8a-octahydronaphthalen-1-yl] (2s)-2-methylbutanoate Chemical compound C([C@H]1[C@@H](C)C=C[C@H]2C[C@@H](O)C[C@@H]([C@H]12)OC(=O)[C@@H](C)CC)CC1C[C@@H](O)CC(=O)O1 LJOOWESTVASNOG-UFJKPHDISA-N 0.000 description 3
- 230000002378 acidificating effect Effects 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 125000004429 atom Chemical group 0.000 description 3
- 239000006143 cell culture medium Substances 0.000 description 3
- 238000012054 celltiter-glo Methods 0.000 description 3
- 239000000460 chlorine Substances 0.000 description 3
- 229940127204 compound 29 Drugs 0.000 description 3
- 229940125878 compound 36 Drugs 0.000 description 3
- 229940125936 compound 42 Drugs 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 125000000753 cycloalkyl group Chemical group 0.000 description 3
- 231100000433 cytotoxic Toxicity 0.000 description 3
- 230000001472 cytotoxic effect Effects 0.000 description 3
- 231100000263 cytotoxicity test Toxicity 0.000 description 3
- 229910052805 deuterium Inorganic materials 0.000 description 3
- 231100000673 dose–response relationship Toxicity 0.000 description 3
- 238000013100 final test Methods 0.000 description 3
- 210000003494 hepatocyte Anatomy 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000010253 intravenous injection Methods 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 239000013642 negative control Substances 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 229940055695 pancreatin Drugs 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 235000011056 potassium acetate Nutrition 0.000 description 3
- 238000003753 real-time PCR Methods 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 210000003462 vein Anatomy 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- UAOUIVVJBYDFKD-XKCDOFEDSA-N (1R,9R,10S,11R,12R,15S,18S,21R)-10,11,21-trihydroxy-8,8-dimethyl-14-methylidene-4-(prop-2-enylamino)-20-oxa-5-thia-3-azahexacyclo[9.7.2.112,15.01,9.02,6.012,18]henicosa-2(6),3-dien-13-one Chemical compound C([C@@H]1[C@@H](O)[C@@]23C(C1=C)=O)C[C@H]2[C@]12C(N=C(NCC=C)S4)=C4CC(C)(C)[C@H]1[C@H](O)[C@]3(O)OC2 UAOUIVVJBYDFKD-XKCDOFEDSA-N 0.000 description 2
- AOSZTAHDEDLTLQ-AZKQZHLXSA-N (1S,2S,4R,8S,9S,11S,12R,13S,19S)-6-[(3-chlorophenyl)methyl]-12,19-difluoro-11-hydroxy-8-(2-hydroxyacetyl)-9,13-dimethyl-6-azapentacyclo[10.8.0.02,9.04,8.013,18]icosa-14,17-dien-16-one Chemical compound C([C@@H]1C[C@H]2[C@H]3[C@]([C@]4(C=CC(=O)C=C4[C@@H](F)C3)C)(F)[C@@H](O)C[C@@]2([C@@]1(C1)C(=O)CO)C)N1CC1=CC=CC(Cl)=C1 AOSZTAHDEDLTLQ-AZKQZHLXSA-N 0.000 description 2
- ABJSOROVZZKJGI-OCYUSGCXSA-N (1r,2r,4r)-2-(4-bromophenyl)-n-[(4-chlorophenyl)-(2-fluoropyridin-4-yl)methyl]-4-morpholin-4-ylcyclohexane-1-carboxamide Chemical compound C1=NC(F)=CC(C(NC(=O)[C@H]2[C@@H](C[C@@H](CC2)N2CCOCC2)C=2C=CC(Br)=CC=2)C=2C=CC(Cl)=CC=2)=C1 ABJSOROVZZKJGI-OCYUSGCXSA-N 0.000 description 2
- SZUVGFMDDVSKSI-WIFOCOSTSA-N (1s,2s,3s,5r)-1-(carboxymethyl)-3,5-bis[(4-phenoxyphenyl)methyl-propylcarbamoyl]cyclopentane-1,2-dicarboxylic acid Chemical compound O=C([C@@H]1[C@@H]([C@](CC(O)=O)([C@H](C(=O)N(CCC)CC=2C=CC(OC=3C=CC=CC=3)=CC=2)C1)C(O)=O)C(O)=O)N(CCC)CC(C=C1)=CC=C1OC1=CC=CC=C1 SZUVGFMDDVSKSI-WIFOCOSTSA-N 0.000 description 2
- GHYOCDFICYLMRF-UTIIJYGPSA-N (2S,3R)-N-[(2S)-3-(cyclopenten-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-3-hydroxy-3-(4-methoxyphenyl)-2-[[(2S)-2-[(2-morpholin-4-ylacetyl)amino]propanoyl]amino]propanamide Chemical compound C1(=CCCC1)C[C@@H](C(=O)[C@@]1(OC1)C)NC([C@H]([C@@H](C1=CC=C(C=C1)OC)O)NC([C@H](C)NC(CN1CCOCC1)=O)=O)=O GHYOCDFICYLMRF-UTIIJYGPSA-N 0.000 description 2
- IUSARDYWEPUTPN-OZBXUNDUSA-N (2r)-n-[(2s,3r)-4-[[(4s)-6-(2,2-dimethylpropyl)spiro[3,4-dihydropyrano[2,3-b]pyridine-2,1'-cyclobutane]-4-yl]amino]-3-hydroxy-1-[3-(1,3-thiazol-2-yl)phenyl]butan-2-yl]-2-methoxypropanamide Chemical compound C([C@H](NC(=O)[C@@H](C)OC)[C@H](O)CN[C@@H]1C2=CC(CC(C)(C)C)=CN=C2OC2(CCC2)C1)C(C=1)=CC=CC=1C1=NC=CS1 IUSARDYWEPUTPN-OZBXUNDUSA-N 0.000 description 2
- WWTBZEKOSBFBEM-SPWPXUSOSA-N (2s)-2-[[2-benzyl-3-[hydroxy-[(1r)-2-phenyl-1-(phenylmethoxycarbonylamino)ethyl]phosphoryl]propanoyl]amino]-3-(1h-indol-3-yl)propanoic acid Chemical compound N([C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)O)C(=O)C(CP(O)(=O)[C@H](CC=1C=CC=CC=1)NC(=O)OCC=1C=CC=CC=1)CC1=CC=CC=C1 WWTBZEKOSBFBEM-SPWPXUSOSA-N 0.000 description 2
- BMQDAIUNAGXSKR-UHFFFAOYSA-N (3-hydroxy-2,3-dimethylbutan-2-yl)oxyboronic acid Chemical compound CC(C)(O)C(C)(C)OB(O)O BMQDAIUNAGXSKR-UHFFFAOYSA-N 0.000 description 2
- IWZSHWBGHQBIML-ZGGLMWTQSA-N (3S,8S,10R,13S,14S,17S)-17-isoquinolin-7-yl-N,N,10,13-tetramethyl-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-amine Chemical compound CN(C)[C@H]1CC[C@]2(C)C3CC[C@@]4(C)[C@@H](CC[C@@H]4c4ccc5ccncc5c4)[C@@H]3CC=C2C1 IWZSHWBGHQBIML-ZGGLMWTQSA-N 0.000 description 2
- WZZBNLYBHUDSHF-DHLKQENFSA-N 1-[(3s,4s)-4-[8-(2-chloro-4-pyrimidin-2-yloxyphenyl)-7-fluoro-2-methylimidazo[4,5-c]quinolin-1-yl]-3-fluoropiperidin-1-yl]-2-hydroxyethanone Chemical compound CC1=NC2=CN=C3C=C(F)C(C=4C(=CC(OC=5N=CC=CN=5)=CC=4)Cl)=CC3=C2N1[C@H]1CCN(C(=O)CO)C[C@@H]1F WZZBNLYBHUDSHF-DHLKQENFSA-N 0.000 description 2
- ONBQEOIKXPHGMB-VBSBHUPXSA-N 1-[2-[(2s,3r,4s,5r)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]oxy-4,6-dihydroxyphenyl]-3-(4-hydroxyphenyl)propan-1-one Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=CC(O)=C1C(=O)CCC1=CC=C(O)C=C1 ONBQEOIKXPHGMB-VBSBHUPXSA-N 0.000 description 2
- UNILWMWFPHPYOR-KXEYIPSPSA-M 1-[6-[2-[3-[3-[3-[2-[2-[3-[[2-[2-[[(2r)-1-[[2-[[(2r)-1-[3-[2-[2-[3-[[2-(2-amino-2-oxoethoxy)acetyl]amino]propoxy]ethoxy]ethoxy]propylamino]-3-hydroxy-1-oxopropan-2-yl]amino]-2-oxoethyl]amino]-3-[(2r)-2,3-di(hexadecanoyloxy)propyl]sulfanyl-1-oxopropan-2-yl Chemical compound O=C1C(SCCC(=O)NCCCOCCOCCOCCCNC(=O)COCC(=O)N[C@@H](CSC[C@@H](COC(=O)CCCCCCCCCCCCCCC)OC(=O)CCCCCCCCCCCCCCC)C(=O)NCC(=O)N[C@H](CO)C(=O)NCCCOCCOCCOCCCNC(=O)COCC(N)=O)CC(=O)N1CCNC(=O)CCCCCN\1C2=CC=C(S([O-])(=O)=O)C=C2CC/1=C/C=C/C=C/C1=[N+](CC)C2=CC=C(S([O-])(=O)=O)C=C2C1 UNILWMWFPHPYOR-KXEYIPSPSA-M 0.000 description 2
- YSUIQYOGTINQIN-UZFYAQMZSA-N 2-amino-9-[(1S,6R,8R,9S,10R,15R,17R,18R)-8-(6-aminopurin-9-yl)-9,18-difluoro-3,12-dihydroxy-3,12-bis(sulfanylidene)-2,4,7,11,13,16-hexaoxa-3lambda5,12lambda5-diphosphatricyclo[13.2.1.06,10]octadecan-17-yl]-1H-purin-6-one Chemical compound NC1=NC2=C(N=CN2[C@@H]2O[C@@H]3COP(S)(=O)O[C@@H]4[C@@H](COP(S)(=O)O[C@@H]2[C@@H]3F)O[C@H]([C@H]4F)N2C=NC3=C2N=CN=C3N)C(=O)N1 YSUIQYOGTINQIN-UZFYAQMZSA-N 0.000 description 2
- TVTJUIAKQFIXCE-HUKYDQBMSA-N 2-amino-9-[(2R,3S,4S,5R)-4-fluoro-3-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-7-prop-2-ynyl-1H-purine-6,8-dione Chemical compound NC=1NC(C=2N(C(N(C=2N=1)[C@@H]1O[C@@H]([C@H]([C@H]1O)F)CO)=O)CC#C)=O TVTJUIAKQFIXCE-HUKYDQBMSA-N 0.000 description 2
- QBWKPGNFQQJGFY-QLFBSQMISA-N 3-[(1r)-1-[(2r,6s)-2,6-dimethylmorpholin-4-yl]ethyl]-n-[6-methyl-3-(1h-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-yl]-1,2-thiazol-5-amine Chemical compound N1([C@H](C)C2=NSC(NC=3C4=NC=C(N4C=C(C)N=3)C3=CNN=C3)=C2)C[C@H](C)O[C@H](C)C1 QBWKPGNFQQJGFY-QLFBSQMISA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 229940126657 Compound 17 Drugs 0.000 description 2
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 2
- 241000700721 Hepatitis B virus Species 0.000 description 2
- 241000282414 Homo sapiens Species 0.000 description 2
- BZLVMXJERCGZMT-UHFFFAOYSA-N Methyl tert-butyl ether Chemical compound COC(C)(C)C BZLVMXJERCGZMT-UHFFFAOYSA-N 0.000 description 2
- OPFJDXRVMFKJJO-ZHHKINOHSA-N N-{[3-(2-benzamido-4-methyl-1,3-thiazol-5-yl)-pyrazol-5-yl]carbonyl}-G-dR-G-dD-dD-dD-NH2 Chemical compound S1C(C=2NN=C(C=2)C(=O)NCC(=O)N[C@H](CCCN=C(N)N)C(=O)NCC(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CC(O)=O)C(N)=O)=C(C)N=C1NC(=O)C1=CC=CC=C1 OPFJDXRVMFKJJO-ZHHKINOHSA-N 0.000 description 2
- 241000233805 Phoenix Species 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- WQDUMFSSJAZKTM-UHFFFAOYSA-N Sodium methoxide Chemical compound [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 description 2
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 description 2
- LNUFLCYMSVYYNW-ZPJMAFJPSA-N [(2r,3r,4s,5r,6r)-2-[(2r,3r,4s,5r,6r)-6-[(2r,3r,4s,5r,6r)-6-[(2r,3r,4s,5r,6r)-6-[[(3s,5s,8r,9s,10s,13r,14s,17r)-10,13-dimethyl-17-[(2r)-6-methylheptan-2-yl]-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-3-yl]oxy]-4,5-disulfo Chemical compound O([C@@H]1[C@@H](COS(O)(=O)=O)O[C@@H]([C@@H]([C@H]1OS(O)(=O)=O)OS(O)(=O)=O)O[C@@H]1[C@@H](COS(O)(=O)=O)O[C@@H]([C@@H]([C@H]1OS(O)(=O)=O)OS(O)(=O)=O)O[C@@H]1[C@@H](COS(O)(=O)=O)O[C@H]([C@@H]([C@H]1OS(O)(=O)=O)OS(O)(=O)=O)O[C@@H]1C[C@@H]2CC[C@H]3[C@@H]4CC[C@@H]([C@]4(CC[C@@H]3[C@@]2(C)CC1)C)[C@H](C)CCCC(C)C)[C@H]1O[C@H](COS(O)(=O)=O)[C@@H](OS(O)(=O)=O)[C@H](OS(O)(=O)=O)[C@H]1OS(O)(=O)=O LNUFLCYMSVYYNW-ZPJMAFJPSA-N 0.000 description 2
- WREOTYWODABZMH-DTZQCDIJSA-N [[(2r,3s,4r,5r)-3,4-dihydroxy-5-[2-oxo-4-(2-phenylethoxyamino)pyrimidin-1-yl]oxolan-2-yl]methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate Chemical compound O[C@@H]1[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O[C@H]1N(C=C\1)C(=O)NC/1=N\OCCC1=CC=CC=C1 WREOTYWODABZMH-DTZQCDIJSA-N 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- YRKCREAYFQTBPV-UHFFFAOYSA-N acetylacetone Chemical compound CC(=O)CC(C)=O YRKCREAYFQTBPV-UHFFFAOYSA-N 0.000 description 2
- 150000001412 amines Chemical class 0.000 description 2
- XRWSZZJLZRKHHD-WVWIJVSJSA-N asunaprevir Chemical compound O=C([C@@H]1C[C@H](CN1C(=O)[C@@H](NC(=O)OC(C)(C)C)C(C)(C)C)OC1=NC=C(C2=CC=C(Cl)C=C21)OC)N[C@]1(C(=O)NS(=O)(=O)C2CC2)C[C@H]1C=C XRWSZZJLZRKHHD-WVWIJVSJSA-N 0.000 description 2
- 239000012298 atmosphere Substances 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical group [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- KGNDCEVUMONOKF-UGPLYTSKSA-N benzyl n-[(2r)-1-[(2s,4r)-2-[[(2s)-6-amino-1-(1,3-benzoxazol-2-yl)-1,1-dihydroxyhexan-2-yl]carbamoyl]-4-[(4-methylphenyl)methoxy]pyrrolidin-1-yl]-1-oxo-4-phenylbutan-2-yl]carbamate Chemical compound C1=CC(C)=CC=C1CO[C@H]1CN(C(=O)[C@@H](CCC=2C=CC=CC=2)NC(=O)OCC=2C=CC=CC=2)[C@H](C(=O)N[C@@H](CCCCN)C(O)(O)C=2OC3=CC=CC=C3N=2)C1 KGNDCEVUMONOKF-UGPLYTSKSA-N 0.000 description 2
- 125000002619 bicyclic group Chemical group 0.000 description 2
- 238000007664 blowing Methods 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Chemical compound BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 229940125904 compound 1 Drugs 0.000 description 2
- 229940125797 compound 12 Drugs 0.000 description 2
- 229940126543 compound 14 Drugs 0.000 description 2
- 229940125758 compound 15 Drugs 0.000 description 2
- 229940126142 compound 16 Drugs 0.000 description 2
- 229940125782 compound 2 Drugs 0.000 description 2
- 229940125810 compound 20 Drugs 0.000 description 2
- 229940126086 compound 21 Drugs 0.000 description 2
- 229940126208 compound 22 Drugs 0.000 description 2
- 229940125833 compound 23 Drugs 0.000 description 2
- 229940125961 compound 24 Drugs 0.000 description 2
- 229940125846 compound 25 Drugs 0.000 description 2
- 229940125851 compound 27 Drugs 0.000 description 2
- 229940126214 compound 3 Drugs 0.000 description 2
- 229940125807 compound 37 Drugs 0.000 description 2
- 229940127573 compound 38 Drugs 0.000 description 2
- 229940125898 compound 5 Drugs 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000001212 derivatisation Methods 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- BRZYSWJRSDMWLG-CAXSIQPQSA-N geneticin Chemical compound O1C[C@@](O)(C)[C@H](NC)[C@@H](O)[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](C(C)O)O2)N)[C@@H](N)C[C@H]1N BRZYSWJRSDMWLG-CAXSIQPQSA-N 0.000 description 2
- JAXFJECJQZDFJS-XHEPKHHKSA-N gtpl8555 Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@@H]1C(=O)N[C@H](B1O[C@@]2(C)[C@H]3C[C@H](C3(C)C)C[C@H]2O1)CCC1=CC=C(F)C=C1 JAXFJECJQZDFJS-XHEPKHHKSA-N 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 239000012442 inert solvent Substances 0.000 description 2
- INQOMBQAUSQDDS-UHFFFAOYSA-N iodomethane Chemical compound IC INQOMBQAUSQDDS-UHFFFAOYSA-N 0.000 description 2
- 238000006317 isomerization reaction Methods 0.000 description 2
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 2
- YNESATAKKCNGOF-UHFFFAOYSA-N lithium bis(trimethylsilyl)amide Chemical compound [Li+].C[Si](C)(C)[N-][Si](C)(C)C YNESATAKKCNGOF-UHFFFAOYSA-N 0.000 description 2
- 201000007270 liver cancer Diseases 0.000 description 2
- 208000014018 liver neoplasm Diseases 0.000 description 2
- 210000005228 liver tissue Anatomy 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 239000012299 nitrogen atmosphere Substances 0.000 description 2
- PIDFDZJZLOTZTM-KHVQSSSXSA-N ombitasvir Chemical compound COC(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@H]1C(=O)NC1=CC=C([C@H]2N([C@@H](CC2)C=2C=CC(NC(=O)[C@H]3N(CCC3)C(=O)[C@@H](NC(=O)OC)C(C)C)=CC=2)C=2C=CC(=CC=2)C(C)(C)C)C=C1 PIDFDZJZLOTZTM-KHVQSSSXSA-N 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 125000004430 oxygen atom Chemical group O* 0.000 description 2
- 229910000160 potassium phosphate Inorganic materials 0.000 description 2
- 235000011009 potassium phosphates Nutrition 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000004467 single crystal X-ray diffraction Methods 0.000 description 2
- 229910000033 sodium borohydride Inorganic materials 0.000 description 2
- 239000012279 sodium borohydride Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 229910052717 sulfur Inorganic materials 0.000 description 2
- 238000010189 synthetic method Methods 0.000 description 2
- HJUGFYREWKUQJT-UHFFFAOYSA-N tetrabromomethane Chemical compound BrC(Br)(Br)Br HJUGFYREWKUQJT-UHFFFAOYSA-N 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 2
- RIOQSEWOXXDEQQ-UHFFFAOYSA-N triphenylphosphine Chemical compound C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 RIOQSEWOXXDEQQ-UHFFFAOYSA-N 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- OZFAFGSSMRRTDW-UHFFFAOYSA-N (2,4-dichlorophenyl) benzenesulfonate Chemical compound ClC1=CC(Cl)=CC=C1OS(=O)(=O)C1=CC=CC=C1 OZFAFGSSMRRTDW-UHFFFAOYSA-N 0.000 description 1
- HUWSZNZAROKDRZ-RRLWZMAJSA-N (3r,4r)-3-azaniumyl-5-[[(2s,3r)-1-[(2s)-2,3-dicarboxypyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl]amino]-5-oxo-4-sulfanylpentane-1-sulfonate Chemical compound OS(=O)(=O)CC[C@@H](N)[C@@H](S)C(=O)N[C@@H]([C@H](C)CC)C(=O)N1CCC(C(O)=O)[C@H]1C(O)=O HUWSZNZAROKDRZ-RRLWZMAJSA-N 0.000 description 1
- UVNPEUJXKZFWSJ-LMTQTHQJSA-N (R)-N-[(4S)-8-[6-amino-5-[(3,3-difluoro-2-oxo-1H-pyrrolo[2,3-b]pyridin-4-yl)sulfanyl]pyrazin-2-yl]-2-oxa-8-azaspiro[4.5]decan-4-yl]-2-methylpropane-2-sulfinamide Chemical compound CC(C)(C)[S@@](=O)N[C@@H]1COCC11CCN(CC1)c1cnc(Sc2ccnc3NC(=O)C(F)(F)c23)c(N)n1 UVNPEUJXKZFWSJ-LMTQTHQJSA-N 0.000 description 1
- KQZLRWGGWXJPOS-NLFPWZOASA-N 1-[(1R)-1-(2,4-dichlorophenyl)ethyl]-6-[(4S,5R)-4-[(2S)-2-(hydroxymethyl)pyrrolidin-1-yl]-5-methylcyclohexen-1-yl]pyrazolo[3,4-b]pyrazine-3-carbonitrile Chemical compound ClC1=C(C=CC(=C1)Cl)[C@@H](C)N1N=C(C=2C1=NC(=CN=2)C1=CC[C@@H]([C@@H](C1)C)N1[C@@H](CCC1)CO)C#N KQZLRWGGWXJPOS-NLFPWZOASA-N 0.000 description 1
- PYRKKGOKRMZEIT-UHFFFAOYSA-N 2-[6-(2-cyclopropylethoxy)-9-(2-hydroxy-2-methylpropyl)-1h-phenanthro[9,10-d]imidazol-2-yl]-5-fluorobenzene-1,3-dicarbonitrile Chemical compound C1=C2C3=CC(CC(C)(O)C)=CC=C3C=3NC(C=4C(=CC(F)=CC=4C#N)C#N)=NC=3C2=CC=C1OCCC1CC1 PYRKKGOKRMZEIT-UHFFFAOYSA-N 0.000 description 1
- BGAJNPLDJJBRHK-UHFFFAOYSA-N 3-[2-[5-(3-chloro-4-propan-2-yloxyphenyl)-1,3,4-thiadiazol-2-yl]-3-methyl-6,7-dihydro-4h-pyrazolo[4,3-c]pyridin-5-yl]propanoic acid Chemical compound C1=C(Cl)C(OC(C)C)=CC=C1C1=NN=C(N2C(=C3CN(CCC(O)=O)CCC3=N2)C)S1 BGAJNPLDJJBRHK-UHFFFAOYSA-N 0.000 description 1
- XWBTZHDDWRNOQH-UHFFFAOYSA-N 3-chloro-2-fluoroaniline Chemical compound NC1=CC=CC(Cl)=C1F XWBTZHDDWRNOQH-UHFFFAOYSA-N 0.000 description 1
- YSEMCVGMNUUNRK-UHFFFAOYSA-N 3-chloro-4-fluoroaniline Chemical compound NC1=CC=C(F)C(Cl)=C1 YSEMCVGMNUUNRK-UHFFFAOYSA-N 0.000 description 1
- POILWHVDKZOXJZ-UHFFFAOYSA-N 4-hydroxypent-3-en-2-one Chemical compound CC(O)=CC(C)=O POILWHVDKZOXJZ-UHFFFAOYSA-N 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical group [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 description 1
- KZBUYRJDOAKODT-UHFFFAOYSA-N Chlorine Chemical compound ClCl KZBUYRJDOAKODT-UHFFFAOYSA-N 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- 208000003322 Coinfection Diseases 0.000 description 1
- 229940126639 Compound 33 Drugs 0.000 description 1
- 229910021595 Copper(I) iodide Inorganic materials 0.000 description 1
- 239000012591 Dulbecco’s Phosphate Buffered Saline Substances 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 206010015548 Euthanasia Diseases 0.000 description 1
- 229920002430 Fibre-reinforced plastic Polymers 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 208000005331 Hepatitis D Diseases 0.000 description 1
- 101000600434 Homo sapiens Putative uncharacterized protein encoded by MIR7-3HG Proteins 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- 229930182816 L-glutamine Natural products 0.000 description 1
- 241000701076 Macacine alphaherpesvirus 1 Species 0.000 description 1
- LOMVENUNSWAXEN-UHFFFAOYSA-N Methyl oxalate Chemical compound COC(=O)C(=O)OC LOMVENUNSWAXEN-UHFFFAOYSA-N 0.000 description 1
- NUGPIZCTELGDOS-QHCPKHFHSA-N N-[(1S)-3-[4-(3-methyl-5-propan-2-yl-1,2,4-triazol-4-yl)piperidin-1-yl]-1-pyridin-3-ylpropyl]cyclopentanecarboxamide Chemical compound C(C)(C)C1=NN=C(N1C1CCN(CC1)CC[C@@H](C=1C=NC=CC=1)NC(=O)C1CCCC1)C NUGPIZCTELGDOS-QHCPKHFHSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 102100037401 Putative uncharacterized protein encoded by MIR7-3HG Human genes 0.000 description 1
- 208000001647 Renal Insufficiency Diseases 0.000 description 1
- PNUZDKCDAWUEGK-CYZMBNFOSA-N Sitafloxacin Chemical compound C([C@H]1N)N(C=2C(=C3C(C(C(C(O)=O)=CN3[C@H]3[C@H](C3)F)=O)=CC=2F)Cl)CC11CC1 PNUZDKCDAWUEGK-CYZMBNFOSA-N 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- SMNRFWMNPDABKZ-WVALLCKVSA-N [[(2R,3S,4R,5S)-5-(2,6-dioxo-3H-pyridin-3-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [[[(2R,3S,4S,5R,6R)-4-fluoro-3,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-hydroxyphosphoryl]oxy-hydroxyphosphoryl] hydrogen phosphate Chemical compound OC[C@H]1O[C@H](OP(O)(=O)OP(O)(=O)OP(O)(=O)OP(O)(=O)OC[C@H]2O[C@H]([C@H](O)[C@@H]2O)C2C=CC(=O)NC2=O)[C@H](O)[C@@H](F)[C@@H]1O SMNRFWMNPDABKZ-WVALLCKVSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 125000006242 amine protecting group Chemical group 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 150000008064 anhydrides Chemical class 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 238000011914 asymmetric synthesis Methods 0.000 description 1
- 125000001584 benzyloxycarbonyl group Chemical group C(=O)(OCC1=CC=CC=C1)* 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000004305 biphenyl Substances 0.000 description 1
- 235000010290 biphenyl Nutrition 0.000 description 1
- YNHIGQDRGKUECZ-UHFFFAOYSA-L bis(triphenylphosphine)palladium(ii) dichloride Chemical compound [Cl-].[Cl-].[Pd+2].C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 YNHIGQDRGKUECZ-UHFFFAOYSA-L 0.000 description 1
- 238000010241 blood sampling Methods 0.000 description 1
- 229910052794 bromium Inorganic materials 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 150000004657 carbamic acid derivatives Chemical class 0.000 description 1
- 150000001721 carbon Chemical group 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 235000011089 carbon dioxide Nutrition 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 150000001793 charged compounds Chemical class 0.000 description 1
- 239000012069 chiral reagent Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 230000006020 chronic inflammation Effects 0.000 description 1
- 230000007882 cirrhosis Effects 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 229940125773 compound 10 Drugs 0.000 description 1
- 229940125877 compound 31 Drugs 0.000 description 1
- 229940126540 compound 41 Drugs 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- TVZPLCNGKSPOJA-UHFFFAOYSA-N copper zinc Chemical compound [Cu].[Zn] TVZPLCNGKSPOJA-UHFFFAOYSA-N 0.000 description 1
- LSXDOTMGLUJQCM-UHFFFAOYSA-M copper(i) iodide Chemical compound I[Cu] LSXDOTMGLUJQCM-UHFFFAOYSA-M 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- XSYZCZPCBXYQTE-UHFFFAOYSA-N cyclodecylcyclodecane Chemical compound C1CCCCCCCCC1C1CCCCCCCCC1 XSYZCZPCBXYQTE-UHFFFAOYSA-N 0.000 description 1
- 125000000582 cycloheptyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C1([H])[H] 0.000 description 1
- NLUNLVTVUDIHFE-UHFFFAOYSA-N cyclooctylcyclooctane Chemical compound C1CCCCCCC1C1CCCCCCC1 NLUNLVTVUDIHFE-UHFFFAOYSA-N 0.000 description 1
- 125000001511 cyclopentyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C1([H])[H] 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 238000009795 derivation Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- RCJVRSBWZCNNQT-UHFFFAOYSA-N dichloridooxygen Chemical compound ClOCl RCJVRSBWZCNNQT-UHFFFAOYSA-N 0.000 description 1
- HQWPLXHWEZZGKY-UHFFFAOYSA-N diethylzinc Chemical compound CC[Zn]CC HQWPLXHWEZZGKY-UHFFFAOYSA-N 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- QUPDWYMUPZLYJZ-UHFFFAOYSA-N ethyl Chemical compound C[CH2] QUPDWYMUPZLYJZ-UHFFFAOYSA-N 0.000 description 1
- SQGRDKSRFFUBBU-UHFFFAOYSA-N ethyl 4-(2-bromo-4-fluorophenyl)-6-(morpholin-4-ylmethyl)-2-(1,3-thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate Chemical compound N1C(C=2SC=CN=2)=NC(C=2C(=CC(F)=CC=2)Br)C(C(=O)OCC)=C1CN1CCOCC1 SQGRDKSRFFUBBU-UHFFFAOYSA-N 0.000 description 1
- 239000011151 fibre-reinforced plastic Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000011737 fluorine Substances 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 125000005843 halogen group Chemical group 0.000 description 1
- 238000002748 hepatotoxicity test Methods 0.000 description 1
- 125000001183 hydrocarbyl group Chemical group 0.000 description 1
- 150000003949 imides Chemical class 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- ZCYVEMRRCGMTRW-YPZZEJLDSA-N iodine-125 Chemical compound [125I] ZCYVEMRRCGMTRW-YPZZEJLDSA-N 0.000 description 1
- 229940044173 iodine-125 Drugs 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 125000003253 isopropoxy group Chemical group [H]C([H])([H])C([H])(O*)C([H])([H])[H] 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 230000000155 isotopic effect Effects 0.000 description 1
- ZLVXBBHTMQJRSX-VMGNSXQWSA-N jdtic Chemical compound C1([C@]2(C)CCN(C[C@@H]2C)C[C@H](C(C)C)NC(=O)[C@@H]2NCC3=CC(O)=CC=C3C2)=CC=CC(O)=C1 ZLVXBBHTMQJRSX-VMGNSXQWSA-N 0.000 description 1
- 201000006370 kidney failure Diseases 0.000 description 1
- 208000006443 lactic acidosis Diseases 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- RENRQMCACQEWFC-UGKGYDQZSA-N lnp023 Chemical compound C1([C@H]2N(CC=3C=4C=CNC=4C(C)=CC=3OC)CC[C@@H](C2)OCC)=CC=C(C(O)=O)C=C1 RENRQMCACQEWFC-UGKGYDQZSA-N 0.000 description 1
- 125000001434 methanylylidene group Chemical group [H]C#[*] 0.000 description 1
- LSEFCHWGJNHZNT-UHFFFAOYSA-M methyl(triphenyl)phosphanium;bromide Chemical compound [Br-].C=1C=CC=CC=1[P+](C=1C=CC=CC=1)(C)C1=CC=CC=C1 LSEFCHWGJNHZNT-UHFFFAOYSA-M 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- XONPDZSGENTBNJ-UHFFFAOYSA-N molecular hydrogen;sodium Chemical compound [Na].[H][H] XONPDZSGENTBNJ-UHFFFAOYSA-N 0.000 description 1
- 125000002950 monocyclic group Chemical group 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 125000003506 n-propoxy group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])O* 0.000 description 1
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000004433 nitrogen atom Chemical group N* 0.000 description 1
- 125000002868 norbornyl group Chemical group C12(CCC(CC1)C2)* 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 150000003833 nucleoside derivatives Chemical class 0.000 description 1
- AICOOMRHRUFYCM-ZRRPKQBOSA-N oxazine, 1 Chemical compound C([C@@H]1[C@H](C(C[C@]2(C)[C@@H]([C@H](C)N(C)C)[C@H](O)C[C@]21C)=O)CC1=CC2)C[C@H]1[C@@]1(C)[C@H]2N=C(C(C)C)OC1 AICOOMRHRUFYCM-ZRRPKQBOSA-N 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 230000002085 persistent effect Effects 0.000 description 1
- ZUOUZKKEUPVFJK-UHFFFAOYSA-N phenylbenzene Natural products C1=CC=CC=C1C1=CC=CC=C1 ZUOUZKKEUPVFJK-UHFFFAOYSA-N 0.000 description 1
- UYWQUFXKFGHYNT-UHFFFAOYSA-N phenylmethyl ester of formic acid Natural products O=COCC1=CC=CC=C1 UYWQUFXKFGHYNT-UHFFFAOYSA-N 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229940068918 polyethylene glycol 400 Drugs 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 230000006340 racemization Effects 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 229930195734 saturated hydrocarbon Natural products 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 125000003003 spiro group Chemical group 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 229940124530 sulfonamide Drugs 0.000 description 1
- 150000003456 sulfonamides Chemical class 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 238000003419 tautomerization reaction Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- IHZAEIHJPNTART-UHFFFAOYSA-N tribromofluoromethane Chemical compound FC(Br)(Br)Br IHZAEIHJPNTART-UHFFFAOYSA-N 0.000 description 1
- PVFOMCVHYWHZJE-UHFFFAOYSA-N trichloroacetyl chloride Chemical compound ClC(=O)C(Cl)(Cl)Cl PVFOMCVHYWHZJE-UHFFFAOYSA-N 0.000 description 1
- 125000001889 triflyl group Chemical group FC(F)(F)S(*)(=O)=O 0.000 description 1
- 229910052722 tritium Inorganic materials 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
- A61K31/549—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame having two or more nitrogen atoms in the same ring, e.g. hydrochlorothiazide
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Virology (AREA)
- Epidemiology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention discloses a methylsulfonyl urea compound. Specifically disclosed are compounds represented by formula (I) and pharmaceutically acceptable salts thereof.
Description
The present application claims priority as follows:
CN2021104946968, filing date: 2021, 5 and 7.
The invention relates to a methylsulfonyl urea compound. In particular to a compound shown in a formula (I) and pharmaceutically acceptable salts thereof.
Hepatitis b virus belongs to the family of hepatiridae. It may cause acute and/or persistent progressive chronic diseases. Hepatitis b virus can also cause many other clinical manifestations in pathological forms, especially chronic inflammation of the liver, cirrhosis and cancer of hepatocytes. In addition, co-infection with hepatitis D can have adverse effects during the course of the disease.
Hepatitis B is a worldwide medical problem, no specific medicine for treating hepatitis B exists worldwide at present, the current first-line medicine for treating hepatitis B is mainly nucleoside medicines and interferon medicines, but the medicines cannot be thoroughly cured, all the medicines need to be taken for a long time, and the problems of renal insufficiency, lactic acidosis and the like exist, so that the development of a novel anti-hepatitis B medicine which is more effective and safer for patients is imperative.
Disclosure of Invention
The invention provides a compound shown as a formula (I) or pharmaceutically acceptable salt thereof,
wherein,
R 1 are each independently selected from halogen, OH, CN, NH 2 、C 1-3 Alkyl and C 1-3 Alkoxy group, the C 1-3 Alkyl groupC 1-3 Alkoxy groups are each independently optionally substituted with 1, 2 or 3 halogens;
m is selected from 0, 1, 2, 3 and 4;
T 1 and T 2 Each independently selected from CH and N;
is a single bond or a double bond;
R 2 selected from H, methyl and F;
ring A is selected from phenyl and C 4-10 Cycloalkyl, the phenyl and C 4-10 Cycloalkyl groups are each independently optionally substituted with 1, 2 or 3R a Substitution;
R a are each independently selected from halogen, OH, CN, NH 2 、C 1-3 Alkyl and C 1-3 Alkoxy group, the C 1-3 Alkyl and C 1-3 Alkoxy groups are each independently optionally substituted with 1, 2 or 3 halogens;
L 1 selected from single bond, -C.ident.C-and-CR 2 ＝；
L 2 Selected from single bonds and methylene groups.
In some aspects of the invention, R is as defined above 1 Are respectively and independently selected from F, cl, br, CN and-CH 3 and-OCH 3 the-CH 3 and-OCH 3 Each independently optionally substituted with one, 2 or 3F, and the other variables are as defined herein.
In some aspects of the invention, R is as defined above 1 Each independently selected from F, cl, br and CN, with the other variables being as defined herein.
In some aspects of the invention, L as described above 1 Selected from-c≡c-, -ch=, -C (F) =, and-C (CH) 3 ) =, other variables are defined as in the present invention.
In some aspects of the inventionR is as described above a Are respectively and independently selected from F, cl, br, CH 3 And OCH 3 The CH is 3 And OCH 3 Each independently optionally substituted with 1, 2 or 3F, and the other variables are as defined herein.
In some aspects of the invention, the structural units described aboveSelected from the group consisting ofThe other variables are as defined herein.
In some aspects of the invention, the structural units described aboveSelected from the group consisting of The other variables are as defined herein.
In some embodiments of the invention, the ring A is selected from phenyl, cyclobutyl, cyclohexyl,The phenyl, cyclobutyl, cyclohexyl,Are each independently optionally substituted with 1, 2 or 3R a Instead, the other variables are as defined herein.
In some embodiments of the invention, the ring A is selected fromThe other variables are as defined herein.
In some aspects of the invention, the structural units described aboveSelected from the group consisting of The other variables are as defined herein.
In some aspects of the invention, the structural units described aboveSelected from the group consisting of The other variables are as defined herein.
In some embodiments of the invention, the above compound, or a pharmaceutically acceptable salt thereof, is selected from the group consisting of:
wherein R is 1 、R 2 、L 2 And m is as defined in any one of the present inventions.
In some embodiments of the invention, the above compound, or a pharmaceutically acceptable salt thereof, is selected from the group consisting of:
wherein R is 1 、R 2 、L 2 And m is as defined in any one of the present inventions.
In some embodiments of the invention, the above compound, or a pharmaceutically acceptable salt thereof, is selected from the group consisting of:
wherein R is 1 、R 2 、L 2 And m is as defined in any one of the present inventions.
Still other embodiments of the present invention are derived from any combination of the variables described above.
The present invention also provides the following compounds, or pharmaceutically acceptable salts thereof, selected from:
the present invention also provides the following compounds, or pharmaceutically acceptable salts thereof, selected from:
the present invention also provides the following compounds, or pharmaceutically acceptable salts thereof, selected from:
the present invention also provides the following compounds, or pharmaceutically acceptable salts thereof, selected from:
the invention also provides application of the compound or the pharmaceutically acceptable salt thereof in preparing medicaments for treating diseases related to hepatitis B virus infection.
In some aspects of the invention, the hepatitis B virus infection-associated disease is chronic hepatitis B.
Definition and description
The following terms and phrases used herein are intended to have the following meanings unless otherwise indicated. A particular term or phrase, unless otherwise specifically defined, should not be construed as being ambiguous or otherwise clear, but rather should be construed in a generic sense. When trade names are presented herein, it is intended to refer to their corresponding commercial products or active ingredients thereof.
The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
The term "pharmaceutically acceptable salts" refers to salts of the compounds of the present application prepared from the compounds found herein to have a particular substituent with a relatively non-toxic acid or base. When relatively acidic functionalities are included in the compounds of the present application, base addition salts may be obtained by contacting such compounds with a sufficient amount of base in pure solution or in a suitable inert solvent. When the compounds of the present invention contain relatively basic functional groups, the acid addition salts may be obtained by contacting such compounds with a sufficient amount of acid in pure solution or in a suitable inert solvent. Certain specific compounds of the invention contain basic and acidic functionalities that can be converted to either base or acid addition salts.
Pharmaceutically acceptable salts of the invention can be synthesized from the parent compound containing an acid or base by conventional chemical methods. In general, the preparation of such salts is as follows: prepared via reaction of these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both.
Unless otherwise indicated, the term "isomer" is intended to include geometric isomers, cis-trans isomers, stereoisomers, enantiomers, optical isomers, diastereomers and tautomers.
The compounds of the invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-) -and (+) -enantiomers, (R) -and (S) -enantiomers, diastereomers, (D) -isomers, (L) -isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of the present invention.
Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of each other.
Unless otherwise indicated, the term "cis-trans isomer" or "geometric isomer" is caused by the inability of a double bond or a single bond of a ring-forming carbon atom to rotate freely.
Unless otherwise indicated, the term "diastereoisomer" refers to stereoisomers of a molecule having two or more chiral centers and having a non-mirror relationship between the molecules.
Unless otherwise indicated, "(+)" means dextrorotation, "(-)" means levorotatory, "(±)" means racemization.
Unless otherwise indicated, with solid wedge bondsAnd a wedge-shaped dotted bondRepresenting the absolute configuration of a solid centre by straight solid keysAnd straight dotted line keyRepresenting the relative configuration of the stereogenic centers, e.g.Indicating that Rx and Ry are co-directional and comprisingTwo configurations.
Unless otherwise specified, when a group has one or more bondable sites, any one or more of the sites of the group may be bonded to other groups by chemical bonds. When the connection mode of the chemical bond is not positioned and the H atoms exist in the connectable site, the number of the H atoms of the site can be correspondingly reduced to be changed into the corresponding valence group along with the number of the connected chemical bond when the chemical bond is connected. The chemical bond of the site and other groups can be a straight solid line bondStraight dotted line keyOr wave linesAnd (3) representing. For example, a straight solid bond in-OCH 3 indicates that it is attached to other groups through an oxygen atom in that group;the straight dashed bonds in (a) represent the attachment to other groups through both ends of the nitrogen atom in the group; by straight broken double bonds Representing the linkage between groups by double bonds, e.g.The double bond of the straight-line indicates that the cyclohexyl group is doubly bonded to other groups.
The compounds of the invention may be present in particular. Unless otherwise indicated, the term "tautomer" or "tautomeric form" refers to the fact that at room temperature, different functional group isomers are in dynamic equilibrium and are capable of rapid interconversion. If tautomers are possible (e.g., in solution), chemical equilibrium of the tautomers can be reached. For example, proton tautomers (also known as proton tautomers) (prototropic tautomer) include interconversions by proton transfer, such as keto-enol isomerisation and imine-enamine isomerisation. Valence isomer (valance tautomer) includes the interconversion by recombination of some of the bond-forming electrons. A specific example of where keto-enol tautomerization is the interconversion between two tautomers of pentane-2, 4-dione and 4-hydroxypent-3-en-2-one.
Unless otherwise indicated, the terms "enriched in one isomer", "enriched in one enantiomer" or "enantiomerically enriched" mean that the content of one isomer or enantiomer is less than 100% and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.
Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or enantiomers. For example, where one isomer or enantiomer is present in an amount of 90% and the other isomer or enantiomer is present in an amount of 10%, the isomer or enantiomer excess (ee value) is 80%.
Optically active (R) -and (S) -isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the invention is desired, it may be prepared by asymmetric synthesis or derivatization with chiral auxiliary wherein the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (e.g., amino) or an acidic functional group (e.g., carboxyl), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereomeric resolution is carried out by conventional methods well known in the art, and then the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomers is usually accomplished by using chromatography employing a chiral stationary phase, optionally in combination with chemical derivatization), for example, to form carbamates from amines.
The compounds of the present invention may contain non-natural proportions of atomic isotopes on one or more of the atoms comprising the compounds. For example, compounds can be labeled with radioisotopes, such as tritium @, for example 3 H) Iodine-125% 125 I) Or C-14% 14 C) A. The invention relates to a method for producing a fibre-reinforced plastic composite For example, deuterium can be substituted for hydrogen to form a deuterated drug, and the bond between deuterium and carbon is stronger than the bond between normal hydrogen and carbon, so that the deuterated drug has the advantages of reducing toxic and side effects, increasing the stability of the drug, enhancing the curative effect, prolonging the biological half-life of the drug and the like compared with the non-deuterated drug. All isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention.
Unless otherwise specified, the term "C 1-3 Alkyl "is used to denote a straight or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C is 1-3 Alkyl includes C 1-2 And C 2-3 Alkyl groups, etc.; it may be monovalent (e.g., methyl), divalent (e.g., methylene), or multivalent (e.g., methine). C (C) 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethylRadical (Et), propyl (including n-propyl and isopropyl), and the like.
Unless otherwise specified, the term "C 1-3 Alkoxy "means those alkyl groups containing 1 to 3 carbon atoms that are attached to the remainder of the molecule through one oxygen atom. The C is 1-3 Alkoxy includes C 1-2 、C 2-3 、C 3 And C 2 Alkoxy groups, and the like. C (C) 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.
Unless otherwise specified, "C 4-10 Cycloalkyl "means a saturated cyclic hydrocarbon group consisting of 4 to 10 carbon atoms, which includes monocyclic, bicyclic, and tricyclic ring systems, wherein bicyclic and tricyclic ring systems include spiro, fused, and bridged rings. The C is 4-10 Cycloalkyl includes C 4-5 、C 5-6 、C 6-7 、C 6-8 、C 6-9 、C 6 、C 7 、C 8 、C 9 Or C 10 Etc.; it may be monovalent, divalent or multivalent. C (C) 4-10 Examples of cycloalkyl groups include, but are not limited to, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, [ 2.2.2.2]Bicyclo octane, [4.4.0 ]]Bicyclodecane, etc.
Unless otherwise specified, the term "halo" or "halogen" by itself or as part of another substituent means a fluorine, chlorine, bromine or iodine atom.
The term "optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.
The term "substituted" means that any one or more hydrogen atoms on a particular atom is substituted with a substituent, which may include deuterium and variants of hydrogen, provided that the valence of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =o), it means that two hydrogen atoms are substituted. Oxygen substitution does not occur on the aromatic group. The term "optionally substituted" means that the substituents may or may not be substituted, and the types and numbers of substituents may be arbitrary on the basis that they can be chemically achieved unless otherwise specified.
When any variable (e.g., R) occurs more than once in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0 to 2R, the group may optionally be substituted with up to two R's, and R's in each case have independent options. Furthermore, combinations of substituents and/or variants thereof are only permissible if such combinations result in stable compounds.
When the number of one linking group is 0, such as- (CRR) 0 -it is meant that the linking group is a single bond.
When one of the variables is selected from a single bond, the two groups to which it is attached are indicated as being directly linked, e.g., when L in A-L-Z represents a single bond, it is indicated that the structure is actually A-Z.
When a substituent is absent, it is meant that the substituent is absent, e.g., X in A-X is absent, meaning that the structure is actually A. When the exemplified linking group does not indicate its linking direction, its linking direction is arbitrary, for example,the linking group L is-M-W-, in which case-M-W-may be a group formed by linking the rings A and B in the same direction as the reading order from left to rightThe ring A and the ring B may be connected in a direction opposite to the reading order from left to right The linking group, the substitutionCombinations of bases and/or variants thereof are permissible only if such combinations result in stable compounds.
The compounds of the present invention may be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, embodiments formed by combining with other chemical synthetic methods, and equivalent alternatives well known to those skilled in the art, preferred embodiments including but not limited to the examples of the present invention.
The compounds of the present invention may be structured by conventional methods well known to those skilled in the art, and if the present invention relates to the absolute configuration of a compound, the absolute configuration may be confirmed by conventional means in the art. For example, single crystal X-ray diffraction (SXRD), the grown single crystal is collected from diffraction intensity data using a Bruker D8 vent diffractometer, and the light source is cukα radiation, scanning:after scanning and collecting the relevant data, the absolute configuration can be confirmed by further analyzing the crystal structure by a direct method (Shellxs 97).
The solvent used in the present invention is commercially available. The invention adopts the following abbreviations: eq represents equivalent, equivalent; m represents mol/L; DMF represents N, N-dimethylformamide; DMSO represents dimethylsulfoxide; etOH stands for ethanol; meOH represents methanol; CBz represents benzyloxycarbonyl, which is a protecting group for an amine; boc represents that t-butoxycarbonyl is an amine protecting group; IPA represents isopropanol and dppf represents 1,1' -bisdiphenylphosphino ferrocene.
Compounds are either prepared according to the general nomenclature of the art or are usedSoftware naming, commercial compounds are referred to by vendor catalog names. Technical effects
As a new type of anti-hepatitis B medicine, the compound has obvious inhibition effect on HBV, has no obvious cytotoxicity in various liver cells, has high bioavailability of oral administration in mice, has high liver-blood ratio of the medicine, and has obvious antiviral effect in an HDI drug effect model.
The present invention is described in detail below by way of examples, but is not meant to be limiting in any way. The present invention has been described in detail herein, and specific embodiments thereof are also disclosed, it will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the invention without departing from the spirit and scope of the invention.
Example 1
The synthetic route is as follows:
step 1: synthesis of Compound 1-b
To a round bottom flask was added compound 1-a (300 g,2.36mol,243.90mL,1 eq), ethylene glycol (292.85 g,4.72mol,263.83mL,2 eq) and toluene (1.5L) under nitrogen followed by p-toluene sulfonic acid (40.63 g,235.92mmol,0.1 eq) and refluxed at 120℃for 12 hours while water was separated by a water separator. The reaction mixture was washed with water (300 mL. Times.2). The aqueous phase was extracted with ethyl acetate (100 mL). The combined organic phases were washed with saturated brine (300 mL. Times.2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was separated by column chromatography on silica gel (petroleum ether: ethyl acetate=100:0 to 50:50) to give compound 1-b.
1 H NMR(400MHz，CDCl 3 )δppm7.72(d，J＝3.1Hz，1H)，7.30-7.08(m，1H)，4.11-4.02(m，2H)，3.99-3.92(m，2H)，1.77(s，3H).
Step 2: synthesis of Compound 1-c
Compound 1-b (20 g,116.81mmol,1 eq) was dissolved in tetrahydrofuran (360 mL), cooled to-78deg.C, and butyllithium (2.5M, 51.40mL,1.1 eq) was slowly added dropwise. The reaction solution was stirred at-78℃for 1 hour, then a solution of carbon tetrabromide (42.61 g,128.49mmol,1.1 eq) in tetrahydrofuran (40 mL) was slowly added dropwise, the reaction solution was stirred at-78℃for 0.5 hour, and the temperature was raised to 0℃and stirred for 0.5 hour. To the reaction mixture was added dropwise a saturated ammonium chloride solution (300 mL), followed by extraction with ethyl acetate (300 mL. Times.3). The combined organic phases were washed with saturated brine (300 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 10/1) to give compound 1-c.
MS m/z(ESI)：249.8[M+1] + .
1 H NMR(400MHz，CDCl 3 )δppm 7.65(s，1H)，4.12-4.07(m，2H)，4.03-3.98(m，2H)，1.79(s，3H).
Step 3: synthesis of Compound 1-d
In a three-necked flask, compound 1-c (49.1 g,196.31mmol,1 eq) and acetone (280 mL) were added, and the mixture was reacted at 25℃with stirring for 2 hours while adding an aqueous hydrochloric acid solution (6M, 196.31mL,6 eq) at 0 ℃. Water (200 mL) was added to the reaction mixture, the mixture was extracted with ethyl acetate (200 mL. Times.3), and the combined organic phases were washed with saturated brine (200 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 1-d.
1 H NMR(400MHz，CDCl 3 )δppm 7.87(s，1H)，2.65(d，J＝0.8Hz，3H).
Step 4: synthesis of Compound 1-e
In a three-necked flask, compound 1-d (22 g,106.76mmol,1 eq) and methanol (300 mL) were added followed by slow addition of sodium methoxide in methanol at 0deg.CSolution (5M, 42.71mL,2 eq) and dimethyl oxalate (18.91 g,160.15mmol,1.5 eq). The mixture was stirred at 70℃for 1 hour. The reaction mixture was cooled to 0℃and water (300 mL) was added thereto, followed by pH=2 to 3 with 2mol/L dilute hydrochloric acid, and the mixture was stirred at 0℃for 30 minutes and then filtered. The cake was washed with methanol (30 mL. Times.3) and dried under reduced pressure. Compound 1-e is obtained. MS m/z (ESI): 291.9[ M+1 ]] + .
1 H NMR(400MHz，DMSO-d 6 )δppm 7.92(s，1H)，6.55(s，1H)，3.69(s，3H).
Step 5: synthesis of Compound 1-f
Into a round bottom flask was charged compound 1-e (65 g,222.52mmol,1 eq), sulfonamide (32.08 g,333.78mmol,19.93mL,1.5 eq) and hydrochloric acid/methanol solution (4M, 500mL,8.99 eq). The mixture was stirred at 70℃for 2 hours. The reaction solution was cooled to 0 ℃, stirred for 1 hour and then filtered. Water (300 mL) was added to the filter cake, stirred for 30 minutes and filtered. To the filter cake was added tert-butyl methyl ether (200 mL), stirred for 30 minutes and filtered. And (5) drying the filter cake under reduced pressure to obtain the compound 1-f. MS m/z (ESI): 351.9[ M+1 ]] + .
1 H NMR(400MHz，DMSO-d 6 )δppm 8.05(s，1H)，6.84(s，1H)，3.81(s，3H).
Step 6: synthesis of Compound 1-g
Compounds 1-f (20 g,56.79mmol,1 eq) and DMF (200 mL) were added under nitrogen to a three-necked flask, dissolved by stirring, and sodium hydrogen (2.95 g,73.82mmol,60% purity, 1.3 eq) was added in portions at 0deg.C. The reaction mixture was stirred at 0deg.C for 30 min, then methyl iodide (24.18 g,170.37mmo1, 10.61mL,3 eq) was added. The mixture was stirred at 50℃for 12 hours. The reaction solution was slowly poured into ice water (200 mL), ph=2 to 3 was adjusted with 1M diluted hydrochloric acid, and the mixture was stirred for 15 minutes and then filtered. The cake was washed with water (50 mL) and methanol (20 mL. Times.2), and dried under reduced pressure. Compound 1-g was obtained.
MS m/z(ESI)：365.7[M+1] + .
1 H NMR(400MHz，DMSO-d 6 )δppm 8.35(s，1H)，7.37(s，1H)，3.95(s，3H)，3.60(s，3H).
Step 7: synthesis of Compounds 1-h
Compound 1-g (50 g,136.53mmol,1 eq) and triethylamine (69.08 g,682.67mmol,95.02mL,5 eq) were added to acetonitrile (250 mL) and water (250 mL), and the mixture was stirred at 50℃for 1 hour. Water (200 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (200 mL. Times.2). The combined organic phases were washed successively with water (100 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound 1-h was obtained.
MS m/z(ESI)：351.8[M+1] + .
Step 8: synthesis of Compound 1-j
Compound 1-i (15 g,77.51mmol,1 eq) was dissolved in methanol (450 mL) and acetic acid (18 mL), reduced iron powder (15.00 g,268.60mmol,3.47 eq) was added in portions, and the reaction mixture was warmed to 65℃and stirred for 2 hours. The reaction solution was cooled to 25 ℃ and filtered, and the filtrate was concentrated under reduced pressure to give a crude product. To the crude product, ethyl acetate (300 mL) was added, followed by washing with 1N aqueous sodium hydroxide (300 mL. Times.2) and saturated brine (300 mL. Times.2), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound 1-j.
1 H NMR(400MHz，CDCl 3 )δ＝6.47-6.41(m，1H)，6.41-6.33(m，1H)，3.71(br s，2H).
Step 9: synthesis of Compound 1-k
Compound 1-h (15 g,42.59mmol,1 eq), compound 1-j (6.97 g,42.59mmol,1 eq), N, N-diisopropylethylamine (16.51 g,127.77mmol,22.26mL,3 eq) and tri-N-propylcyclophosphatemic anhydride in 50% ethyl acetate (40.66 g,63.89mmol,38.00mL,1.5 eq) were dissolved in dichloromethane (450 mL) at 0deg.C, the reaction was replaced three times with nitrogen, and then warmed to 25deg.C and stirred for reaction for 12 hours. Water (500 mL) was added to the reaction mixture, the mixture was extracted with methylene chloride (500 mL. Times.2), and the organic phases were combined, washed with saturated brine (500 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 10/3) to give compound 1-k.
MS m/z(ESI)：498.7[M+1] + .
Step 10: synthesis of Compound 1-l
Compound 1-k (6.87 g,13.80mmol,1 eq) was dissolved in ethanol (100 mL), the reaction solution was cooled to 0deg.C, then sodium borohydride (1.04 g,27.61mmol,2 eq) was added slowly in portions, and the reaction was stirred for 2 hours. To the reaction solution, 1M aqueous hydrochloric acid solution was slowly added to adjust ph=1 to 2, water (100 mL) was further added, and the mixture was stirred for 5 minutes and then filtered. Methanol (20 mL) was added to the filter cake and the mixture was slurried for 30 minutes, filtered, and the filter cake was dried in vacuo to give Compound 1-l.
MS m/z(ESI)：502.8[M+1] + .
Step 11: synthesis of Compound 1-o
2, 6-tetramethylpiperidine (498.33 mg,3.53mmol, 598.95. Mu.L, 1.2 eq) was dissolved in tetrahydrofuran (6 mL), the system was cooled to-30℃and n-butyllithium (2.5M, 1.4mL,1.19 eq) was added dropwise and the reaction stirred at-30℃for 0.5 hours. Then the reaction was cooled to-78℃and a solution of compound 1-n (787.81 mg,2.94mmol,1 eq) in tetrahydrofuran (6 mL) was added dropwise, and stirring was carried out at-78℃for 30min, followed by addition of compound 1-m (0.5 g,2.94mmol, 467.29. Mu.L, 1 eq) in tetrahydrofuran (6 mL). The reaction was slowly warmed to 25℃and stirred at 25℃for 12 hours. The reaction system was cooled to 0℃and saturated ammonium chloride solution (5 mL) was added dropwise, the reaction solution was stirred at 0℃for 1 hour, the filtrate was concentrated under reduced pressure after filtration, water (20 mL) and ethyl acetate (20 mL) were added for extraction, and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product. The crude product was purified by flash column chromatography (eluent: ethyl acetate/petroleum ether, gradient: 0-20%). Compound 1-o is obtained.
1 H NMR(400MHz，CDCl 3 )δ＝5.08(s，1H)，4.13(q，J＝7.1Hz，2H)，3.18-3.09(m，1H)，2.49(tt，J＝3.7，10.9Hz，1H)，2.40-2.33(m，1H)，2.20(dt，J＝3.9，12.6Hz，1H)，2.12-1.99(m，3H)，1.72-1.58(m，2H)，1.30-1.23(m， 15H).
Step 12: synthesis of Compound 1-p
Under nitrogen, 1-l (100 mg, 199.30. Mu. Mol,1 eq), 1-o (87.95 mg, 298.95. Mu. Mol,1.5 eq), sodium carbonate (63.37 mg, 597.90. Mu. Mol,3 eq), dioxane (3 mL) and water (0.5 mL) were added to a thumb flask, followed by tetrakis (triphenylphosphine) palladium (23.03 mg, 19.93. Mu. Mol,0.1 eq). The reaction was stirred at 90℃for 3 hours. Water (10 mL) and ethyl acetate (10 mL) were added to the reaction mixture to extract, and the organic phase was concentrated under reduced pressure to give a crude product. The crude product is purified by column chromatography (eluent: ethyl acetate/petroleum ether: 0-50%). Compound 1-p is obtained.
MS m/z(ESI)：589.0[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.71-7.61(m，2H)，7.57(s，1H)，6.37(s，1H)，5.04-4.98(m，1H)，4.48(dd，J＝3.3，11.8Hz，1H)，4.18-4.12(m，2H)，2.99-2.92(m，1H)，2.75(s，3H)，2.60-2.20(m，6H)，2.07(td，J＝3.6，12.5Hz，2H)，1.67-1.56(m，2H)，1.27-1.24(m，3H).
Step 13: synthesis of Compound 1
Compound 1-p (50 mg, 84.88. Mu. Mol,1 eq) was dissolved in methanol (1 mL), and a solution of sodium hydroxide (10.18 mg, 254.64. Mu. Mol,3 eq) in water (1 mL) was added. The reaction was stirred at 25℃for 1 hour. The reaction solution was added with 2M diluted hydrochloric acid to adjust ph=1 to 3, extracted with ethyl acetate (20 mL), and the organic phase was concentrated under reduced pressure to obtain a crude product. The crude product is purified by a preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile ]; acetonitrile%: 40% -70%,7 min) to obtain compound 1.
MS m/z(ESI)：561.2[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.72-7.61(m，2H)，7.59-7.56(m，1H)，7.58(s，1H)，6.38(s，1H)，5.02(br dd，J＝3.2，11.7Hz，1H)，4.48(dd，J＝3.1，11.8Hz，1H)，2.97(br d，J＝13.9Hz，1H)，2.76(s，3H)，2.64-2.55(m，1H)，2.52-2.45(m，1H)，2.43-2.29(m，3H)，2.28-2.18(m，1H)，2.14-2.06(m，2H)，1.72-1.54(m，2H).
Step 14: synthesis of Compounds 2,3,4 and 5
Compound 1 (90 mg, 160.42. Mu. Mol,1 eq) was subjected to SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 μm); mobile phase: [0.1% NH) 3 H 2 O IPA]:50% -50%) to give a mixture of compounds 3 and 4, compound 2 and compound 5. The mixture of compounds 3 and 4 was subjected to a second SFC (column: DAICEL CHIRALPAK IG (250 mm. Times.30 mm,10 μm): mobile phase: [0.1% NH 3 H 2 O IPA]:60% -60%) to give compound 3 and compound 4.
Compound 2: MS m/z (ESI): 560.9[ M+1 ]] + SFC analysis method (column: chiralpak IG-3.50 x 4.6mmI.D.,3 μm; mobile phase: isopropanol (0.05% diethylamine; flow rate: 4 mL/min), SFC retention time: 0.513 min, ee% = 100%.
1 H NMR(400MHz，CD 3 OD)δ＝7.59-7.50(m，2H)，7.46(s，1H)，6.26(s，1H)，4.90(br dd，J＝3.3，11.7Hz，1H)，4.35(dd，J＝3.3，11.8Hz，1H)，2.85(br d，J＝14.0Hz，1H)，2.64(s，3H)，2.46(br t，J＝10.6Hz，1H)，2.36(br d，J＝13.4Hz，1H)，2.31-2.17(m，3H)，2.14-2.05(m，1H)，2.01-1.93(m，2H)，1.60-1.47(m，2H).
Compound 3: MS m/z (ESI): 560.9[ M+1 ]] + SFC retention time: 0.861 min, ee% = 100%.
1 H NMR(400MHz，CD 3 OD)δ＝7.59-7.50(m，2H)，7.46(s，1H)，6.26(s，1H)，4.90(dd，J＝3.3，11.7Hz，1H)，4.36(dd，J＝3.3，11.8Hz，1H)，2.85(br d，J＝13.9Hz，1H)，2.64(s，3H)，2.50-2.42(m，1H)，2.40-2.32(m，1H)，2.29-2.16(m，3H)，2.14-2.05(m，1H)，2.02-1.94(m，2H)，1.59-1.46(m，2H).
Compound 4: MS m/z (ESI): 561.0[ M+1 ]] + SFC retention time: 2.802 minutes, ee% = 100%.
1 H NMR(400MHz，CD 3 OD)δ＝7.59-7.51(m，2H)，7.46(s，1H)，6.25(s，1H)，4.90(dd，J＝3.3，11.8Hz，1H)，4.36(dd，J＝3.1，11.7Hz，1H)，2.86(br d，J＝14.1Hz，1H)，2.64(s，3H)，2.47-2.41(m，1H)，2.40-2.33(m，1H)，2.29-2.17(m，3H)，2.13-2.03(m，1H)，2.00-1.93(m，2H)，1.61-1.43(m，2H).
Compound 5: MS m/z (ESI): 560.9[ M+1 ]] + SFC retention time: 3.495 minutes, ee% = 100%.
1 H NMR(400MHz，CD 3 OD)δ＝7.71-7.62(m，2H)，7.58(s，1H)，6.38(s，1H)，5.02(dd，J＝3.4，11.8Hz，1H)，4.47(dd，J＝3.2，11.7Hz，1H)，2.97(br d，J＝13.8Hz，1H)，2.76(s，3H)，2.62-2.54(m，1H)，2.48(br d，J＝13.5Hz，1H)，2.43-2.29(m，3H)，2.25-2.15(m，1H)，2.13-2.05(m，2H)，1.71-1.58(m，2H).
Example 2
The synthetic route is as follows:
step 1: synthesis of Compound 6-b
Compound 1-l (50 mg, 99.65. Mu. Mol,1 eq) was dissolved in N, N-dimethylformamide (1.4 mL) and triethylamine (1.4 mL) under nitrogen, and Compound 6-a (86.79 mg, 498.25. Mu. Mol,5 eq) and bis (triphenylphosphine) palladium dichloride (3.50 mg, 4.98. Mu. Mol,0.05 eq) and cuprous iodide (3.80 mg, 19.93. Mu. Mol,0.2 eq) were further added thereto, and the reaction mixture was heated to 80℃and stirred for 12 hours. To the reaction solution was added water (10 mL) and ethyl acetate (10 mL. Times.3) for extraction, and the organic phase was washed with 1M diluted hydrochloric acid (10 mL. Times.2) and saturated brine (10 mL. Times.2) in this order, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 2/1) to give compound 6-b.
MS m/z(ESI)：594.9[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝8.05(d，J＝8.5Hz，2H)，7.98(s，1H)，7.70-7.62(m，4H)，5.05(dd，J＝3.1，11.9Hz，1H)，4.48(dd，J＝3.0，12.1Hz，1H)，4.39(d，J＝7.1Hz，2H)，2.75(s，3H)，2.45(td，J＝3.2，14.3Hz，1H)，2.36-2.27(m，1H)，1.42-1.38(m，3H).
Step 2: synthesis of Compound 6
Compound 6-b (40 mg, 67.22. Mu. Mol,1 eq) was dissolved in methanol (1 mL) and water (1 mL), and sodium hydroxide (13.44 mg, 336.11. Mu. Mol,5 eq) was added thereto and the mixture was stirred at 25℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative high performance liquid chromatography (column type: phenomenex Gemini-NXC18X30mmX3μm; mobile phase: water (0.225% formic acid) -acetonitrile;: acetonitrile: 45% -75%,7 min) to give compound 6.
MS m/z(ESI)：567.1[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝8.04(d，J＝8.5Hz，2H)，7.97(s，1H)，7.70-7.60(m，4H)，5.04(dd，J＝3.0，12.0Hz，1H)，4.48(dd，J＝3.0，12.0Hz，1H)，2.74(s，3H)，2.48-2.24(m，2H).
Example 3
The synthetic route is as follows:
step 1: synthesis of Compound 7-c:
compound 1-l (9.3 g,18.54mmol,1 eq) by SFC (column type: REGIS (s, s) WHELK-O1 (250 mm. Times.50 mm,10 μm); mobile phase: [0.1% NH 3 H 2 O，EtOH]Percent: 25% -25%) and collecting the second peak to give compound 7-c.
MS m/z(ESI)：500.8[M+1] + SFC analysis method (column type (S, S) -Whelk-O1 100X 4.6mm I.D.,3 μm; mobile phase: ethanol (0.05% diethylamine; flow rate: 2.8 mL/min); retention time: 2.990 minutes; ee% = 100%.
Step 2: synthesis of Compound 7-a
To the reaction flask were added compound 1-m (2 g,11.75mmol,1.87mL,1 eq), tribromofluoromethane (3.82 g,14.10mmol,1.2 eq), triphenylphosphine (3.70 g,14.10mmol,1.2 eq) and tetrahydrofuran (60 mL). Diethyl zinc (1M, 14.10mL,1.2 eq) was added dropwise at 20deg.C. The reaction solution was stirred at 20℃for 1 hour. Methanol (50 mL) was added to the reaction mixture, and the mixture was stirred for 20 minutes and then concentrated under reduced pressure. The residue was separated by automatic column chromatography COMBI-FLASH (eluent: petroleum ether: ethyl acetate=100:0 to 10:1) to give compound 7-a.
1 H NMR(400MHz，CDCl 3 )δ＝4.08-4.02(m，2H)，2.72-2.68(m，1H)，2.39-2.34(m，2H)，1.95-1.81(m，4H)，1.52-1.49(m，2H)，1.21-1.18(m，3H).
Step 3: synthesis of Compound 7-b
Compound 7-a (2.3 g,8.68mmol,1 eq) was dissolved in 1, 4-dioxane (30 mL) and then pinacol borate (4.41 g,17.35mmol,2 eq), potassium acetate (2.55 g,26.03mmol,3 eq) and Pd (dppf) Cl were added 2 ·CH 2 Cl 2 (708.46 mg, 867.54. Mu. Mol,0.1 eq). After three nitrogen substitutions, the mixture was heated to 90℃and stirred for 16 hours. To the reaction solution was added water (50 mL), extracted with ethyl acetate (30 mL. Times.2), and the combined organic phases were washed with saturated aqueous sodium chloride solution (50 mL. Times.1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated by automatic column chromatography COMBI-FLASH (eluent: petroleum ether: ethyl acetate=100:0 to 10:1) to give compound 7-b.
1 H NMR(400MHz，CDCl 3 )δ＝4.08-4.02(m，2H)，2.92-2.88(m，2H)，2.43-2.37(m，1H)，1.97-1.92(m，2H)，1.97-1.92(m，2H)，1.52-1.49(m，2H)，1.21-1.18(m，15H).
Step 4: synthesis of Compound 7-d
In a thumb flask under nitrogen, compound 7-c (150.00 mg, 298.95. Mu. Mol,1 eq), 7-b (466.64 mg,1.49mmol,5 eq), sodium carbonate (158.43 mg,1.49mmol,5 eq), 1, 4-dioxane (8 mL) and water (1.5 mL) were added followed by Pd (PPh) 3 ) 4 (34.55 mg, 29.90. Mu. Mol,0.1 eq). The mixture was stirred at 100℃for 16 hours. After the reaction was cooled to about 25 ℃, water (10 mL) was added to the reaction mixture, the pH was adjusted to 1 to 2 with 1M diluted hydrochloric acid, and extraction was performed with ethyl acetate (15 ml×3). The combined organic phases were washed with saturated brine (30 mL) and concentrated under reduced pressure. The residue was isolated by automatic column chromatography COMBI-FLASH (gradient elution: petroleum ether: ethyl acetate=100:0 to 70:30) to give compound 7-d.
MS m/z(ESI)：607.0[M+1] + .
1H NMR(400MHz，DMSO-d 6 )δ＝7.97-7.90(m，2H)，7.73(q，J＝5.5Hz，2H)，4.97(br t，J＝9.8Hz，1H)，4.43(dd，J＝2.5，12.0Hz，1H)，4.10-4.06(m，2H)，2.78(br d，J＝14.1Hz，1H)，2.63(s，3H)，2.57(td，J＝3.8，10.4Hz，1H)，2.42-2.35(m，1H)，2.24-2.14(m，2H)，2.13-2.05(m，1H)，1.87-1.77(m，1H)，1.76-1.66(m，1H)，1.64-1.57(m，1H)，1.56-1.43(m，3H)，1.39-1.27(m，1H)，1.21-1.18(m，3H).
Step 5: synthesis of Compound 7
Compounds 7-d (100 mg, 164.73. Mu. Mol,1 eq) and tetrahydrofuran (0.5 mL) were added to the thumb flask followed by aqueous sodium hydroxide (2M, 0.5mL,6.07 eq). The mixture was stirred at 25℃for 1 hour. Water (5 mL) was added to the reaction mixture, pH=1 to 2 was adjusted with 1M diluted hydrochloric acid, and extraction was performed with ethyl acetate (15 mL. Times.3). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated by preparative high performance liquid chromatography (column form Phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase [ water (0.225% formic acid) -acetonitrile ]; acetonitrile%: 40% -70%,7 min) to give compound 7.
MS m/z(ESI)：579.0[M+1] + .
1H NMR(400MHz，DMSO-d 6 )δ＝7.99-7.88(m，2H)，7.78-7.68(m，2H)，4.98(br t，J＝9.6Hz，1H)，4.43(dd，J＝2.5，12.0Hz，1H)，2.77(br d，J＝13.8Hz，1H)，2.63(s，3H)，2.55(br s，1H)，2.49-2.45(m，1H)，2.43-2.35(m，1H)，2.24-2.11(m，2H)，2.08-2.04(m，1H)，1.98(br dd，J＝4.1，8.1Hz，2H)，1.49(q，J＝11.6Hz，2H).
Example 4
The synthetic route is as follows:
step 1: synthesis of Compound 8-c
Compound 8-a (500 mg,2.97mmol,1 eq) was dissolved in tetrahydrofuran (15 mL), the reaction mixture was cooled to-78℃and then LiHMDS (1M, 2.97mL,1 eq) was slowly added dropwise thereto, the reaction was stirred at-78℃for 0.5 hours, then a solution of compound 8-b (1.06 g,2.97mmol,1 eq) in tetrahydrofuran (15 mL) was slowly added dropwise thereto, the reaction was stirred at-78℃for 0.5 hours and then the temperature was raised to 25℃for 1 hour. To the reaction mixture was slowly added water (20 mL), followed by extraction with ethyl acetate (20 mL. Times.3), and the combined organic phases were washed with saturated brine (10 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 10/1) to give compound 8-c.
1 H NMR(400MHz，CDCl 3 )δ＝5.62-5.39(m，1H)，3.71(d，J＝3.5Hz，3H)，3.16-3.02(m，1H)，2.92(d，J＝16.6Hz，2H)，2.62-2.50(m，2H)，2.48-2.38(m，2H).
Step 2: synthesis of Compound 8-d
Compound 8-c (400 mg,1.33mmol,1 eq), bis-pinacolato borate (405.96 mg,1.60mmol,1.2 eq), potassium acetate (392.24 mg,4.00mmol,3 eq) and 1, 1-bis (diphenylphosphine) ferrocene palladium chloride (97.48 mg, 133.22. Mu. Mol,0.1 eq) were dissolved in 1, 4-dioxane (7 mL) under nitrogen, and the reaction system was replaced three times with nitrogen and then warmed to 90℃and stirred for 3 hours. The reaction solution was concentrated directly under reduced pressure, and the crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 10/1) to give compound 8-d.
1 H NMR(400MHz，CDCl 3 )δ＝7.12-6.70(m，1H)，3.68(s，3H)，3.10-2.99(m，1H)，2.68-2.53(m，2H)，2.53-2.32(m，4H)，1.27(s，12H).
Step 3: synthesis of Compound 8-e
Under nitrogen atmosphere, compound 7-c (100 mg, 199.30. Mu. Mol,1 eq), compound 8-d (166.31 mg, 597.90. Mu. Mol,3 eq), potassium phosphate (126.91 mg, 597.90. Mu. Mol,3 eq) and 1, 1-bis (diphenylphosphine) ferrocene palladium chloride (14.58 mg, 19.93. Mu. Mol,0.1 eq) were dissolved in 1, 4-dioxane (1.4 mL) and water (0.4 mL), and the reaction system was replaced three times with nitrogen and then warmed to 80℃and stirred for 3 hours. The reaction solution was concentrated directly under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 2/1) to give compound 8-e.
MS m/z(ESI)：573.2[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.70-7.60(m，5H)，6.43-6.22(m，1H)，5.01-4.97(m，1H)，4.47-4.44(m，1H)，3.68(s，3H)，2.79(s，1H)，2.73(s，3H)，2.49-2.44(m，3H)，2.42-2.36(m，2H)，2.28(br d，J＝14.3Hz，2H).
Step 4: synthesis of Compound 8-f
Compound 8-e (50.00 mg, 87.26. Mu. Mol,1 eq) was dissolved in methanol (3 mL), the reaction system was replaced with nitrogen three times, wet palladium on carbon (92.86 mg, 87.26. Mu. Mol,10% purity, 1 eq) was added, and after replacing with hydrogen three times, the reaction mixture was stirred at 25℃under a hydrogen atmosphere (15 Psi) for 3 hours. The reaction solution was filtered through celite, and concentrated under reduced pressure to give compound 8-f.
MS m/z(ESI)：575.2[M+1] + .
Step 5: synthesis of Compound 8
Compound 8-f (15.00 mg, 26.08. Mu. Mol,1 eq) was dissolved in methanol (1 mL) and water (1 mL), and sodium hydroxide (5.22 mg, 130.42. Mu. Mol,5 eq) was added thereto and the mixture was stirred at 25℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product is purified by a preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile ]; acetonitrile%: 40% -70%,7 min) to obtain compound 8.
MS m/z(ESI)：561.2[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.70-7.58(m，2H)，7.44(s，1H)，4.94(br d，J＝3.3Hz，1H)，4.51-4.39(m，1H)，3.02(quin，J＝8.3Hz，1H)，2.73(s，3H)，2.66-2.58(m，1H)，2.52-2.09(m，10H).
Example 5
The synthetic route is as follows:
step 1: synthesis of Compound 9-b
2, 6-tetramethylpiperidine (2.38 g,16.88mmol,2.87mL,1.2 eq) was dissolved in tetrahydrofuran (24 mL), the system was cooled to-30deg.C and n-butyllithium (2.5M, 6.70mL,1.19 eq) was added. The reaction was stirred at-30℃for 0.5 hour, the reaction was cooled to-78℃and a solution of compound 1-n (3.77 g,14.07mmol,1 eq) in tetrahydrofuran (24 mL) was added, and stirring was carried out at-78℃for 30 minutes and compound 9-a (2 g,14.07mmol,1.87mL,1 eq) in tetrahydrofuran (24 mL) was added. The reaction was slowly warmed to 25℃and stirred at 25℃for 12 hours. The system was cooled to 0deg.C, saturated ammonium chloride solution (20 mL) was added dropwise, the reaction solution was stirred at 0deg.C for 1 hour, the filtrate was concentrated under reduced pressure after the reaction solution was filtered, water (20 mL), ethyl acetate (20 mL. Times.2) was added for extraction, and the organic phase was washed with saturated brine (20 mL) and concentrated under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=100:0-90:10) to give compound 9-b.
1 H NMR(400MHz，CDCl 3 )δ＝5.25-5.15(m，1H)，4.27-4.08(m，2H)，3.23-3.08(m，4H)，3.00-2.93(m，1H)，1.30-1.28(m，3H)，1.25(d，J＝1.5Hz，12H).
Step 2: synthesis of Compound 9-c
Compound 7-c (100.00 mg, 199.30. Mu. Mol,1 eq) and compound 9-b (159.13 mg, 597.90. Mu. Mol,3 eq), sodium carbonate (63.37 mg, 597.90. Mu. Mol,3 eq), 1, 4-dioxane (2 mL) and water (0.3 mL) were added to a thumb flask under nitrogen, followed by tetrakis (triphenylphosphine) palladium (23.03 mg, 19.93. Mu. Mol,0.1 eq). The reaction was stirred at 90℃for 12 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=100:0-50:50) to give compound 9-c.
MS m/z(ESI)：561.0[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.71-7.62(m，2H)，7.55(s，1H)，6.47(br s，1H)，5.01(br d，J＝9.3Hz，1H)，4.47(dd，J＝3.0，11.8Hz，1H)，4.19(q，J＝7.1Hz，2H)，3.33(s，1H)，3.22-3.10(m，4H)，2.75(s，3H)，2.43-2.30(m，2H)，1.29(t，J＝7.0Hz，3H).
Step 3: synthesis of Compound 9
Compound 9-c (25.00 mg, 44.56. Mu. Mol,1 eq) was dissolved in methanol (2 mL) and a solution of sodium hydroxide (5.35 mg, 133.68. Mu. Mol,3 eq) in water (2 mL) was added. The reaction was stirred at 25℃for 30 minutes. The reaction solution was added with 2M diluted hydrochloric acid to adjust ph=2 to 3, extracted with ethyl acetate (10 ml×3), and the organic phase was concentrated under reduced pressure to obtain a crude product. The crude product was purified by preparative high performance liquid chromatography (column: welch Xtimate C18X 40mm X3 μm; mobile phase [ water (0.075% trifluoroacetic acid) -acetonitrile ]; acetonitrile%: 45% -75%,8 min) to give compound 9.
MS m/z(ESI)：533.1[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.72-7.62(m，2H)，7.56(s，1H)，6.49(t，J＝2.3Hz，1H)，5.01(br d，J＝9.3Hz，1H)，4.47(dd，J＝3.4，11.9Hz，1H)，3.30(s，1H)，3.20(br s，2H)，3.14(br d，J＝8.5Hz，2H)，2.75(s，3H)，2.44-2.25(m，2H).
Example 6
The synthesis method comprises the following steps:
step 1: synthesis of Compound 10-b
2, 6-tetramethylpiperidine (9.20 g,65.14mmol,11.06mL,1.2 eq) was dissolved in tetrahydrofuran (120 mL) under nitrogen protection, after the reaction system was cooled to-30 ℃, n-butyllithium (2.5M, 26.05mL,1.2 eq) was slowly added dropwise, the reaction solution was stirred at-30℃for 0.5 hours and then cooled to-78℃and then a tetrahydrofuran (120 mL) solution of compound 1-n (14.54 g,54.28mmol,1 eq) was slowly added dropwise, the reaction solution was stirred at-78℃for 0.5 hours and then a tetrahydrofuran (120 mL) solution of compound 10-a (10 g,54.28mmol,1 eq) was added, and the reaction solution was heated to 25℃and stirred for 12 hours. After the completion of the reaction, the reaction mixture was cooled to 0℃and then a saturated ammonium chloride solution (100 mL) was slowly added dropwise thereto, and the reaction mixture was stirred at 0℃for 10 minutes. The reaction mixture was filtered and dried by spin-drying, water (60 mL) was added, the mixture was extracted with ethyl acetate (60 mL. Times.3), and the organic phase was washed with saturated brine (60 mL. Times.3), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate under reduced pressure to obtain crude product. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 10/1) to give compound 10-b.
1 H NMR(400MHz，CDCl 3 )δ＝5.05(s，1H)，4.18-4.08(m，2H)，3.14(br dd，J＝1.4，13.4Hz，1H)，2.34-2.18(m，4H)，2.05-1.95(m，2H)，1.91-1.82(m，2H)，1.28-1.24(m，15H)，1.20-1.09(m，2H).
Step 2: synthesis of Compound 10-d and Compound 10-e
Compound 7-c (200 mg, 398.60. Mu. Mol,1 eq), compound 10-b (614.29 mg,1.99mmol,5 eq) and N, N-diisopropylethylamine (154.55 mg,1.20mmol, 208.29. Mu.L, 3 eq) were dissolved in 1, 4-dioxane (1.5 mL) and water (1.5 mL) under nitrogen, and bis (triphenylphosphine) palladium dichloride (27.98 mg, 39.86. Mu. Mol,0.1 eq) was added thereto, and the reaction mixture was heated to 100℃and stirred for 12 hours. The reaction solution was concentrated directly under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 2/1) to give compound 10-c. Then SFC (column type: DAICEL CHIRALPAK IG (250 mm. Times.30 mm,10 μm), mobile phase: [0.1% NH ] 3 H 2 O，EtOH]Percent: 60% -60%) to give compound 10-d and compound 10-e.
Compound 10-c: MS m/z (ESI): 603.1[ M+1 ]] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.71-7.59(m，2H)，7.54(s，1H)，6.33(s，1H)，4.99(dd，J＝3.5，11.5Hz，1H)，4.46(dd，J＝3.4，11.7Hz，1H)，4.14-4.11(m，2H)，2.99(br d，J＝13.8Hz，1H)，2.74(s，3H)，2.44-2.24(m，6H)，2.15-2.02(m，2H)，1.93(br d，J＝12.8Hz，2H)，1.25(d，J＝1.0Hz，3H)，1.17-1.08(m，2H).
SFC analysis method: column type: chiralpak IG-3.6mm I.D.,3 μm; mobile phase: ethanol (0.05% diethylamine); flow rate: 4mL/min.
Compound 10-d: MS m/z (ESI): 603.1[ M+1 ]] + SFC retention time: 1.982 minutes; ee% = 100%.
Compound 10-e: MS m/z (ESI): 603.1[ M+1 ]] + SFC retention time: 4.655 minutes; ee% = 99.5%.
Step 3: synthesis of Compound 10
Compound 10-c (70 mg, 116.07. Mu. Mol,1 eq), sodium hydroxide (2M, 290.17. Mu.L, 5 eq), tetrahydrofuran (3 mL) and water (1 mL) were added to a eggplant-shaped bottle, and the reaction solution was stirred at 20℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product is purified by preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile;: 25% -95% acetonitrile, 7 min) to obtain compound 10.
MS m/z(ESI)：575.1[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.78-7.48(m，3H)，6.34(br s，1H)，5.01(br d，J＝9.5Hz，1H)，4.47(br d， J＝9.0Hz，1H)，3.01(br d，J＝14.1Hz，1H)，2.75(s，3H)，2.51-1.84(m，10H)，1.42-1.05(m，2H).
Step 4: synthesis of Compound 11
Compound 10-d (260 mg, 431.11. Mu. Mol,1 eq) was dissolved in methanol (2 mL) and water (2 mL), and sodium hydroxide (86.21 mg,2.16mmol,5 eq) was added thereto and the mixture was stirred at 25℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product is purified by preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile;: acetonitrile%: 40% -70%,7 min) to obtain compound 11.
MS m/z(ESI)：575.1[M+1] + .
SFC analysis method (column: chiralPak AS-3X 4.6mm I.D.,3 μm; mobile phase: isopropanol (0.05% diethylamine; flow rate: 2.5 mL/min); SFC retention time: 5.559 minutes; ee% = 100%.
1 H NMR(400MHz，CD 3 OD)δ＝7.70-7.60(m，2H)，7.54(s，1H)，6.33(s，1H)，4.99(br dd，J＝3.3，11.8Hz，1H)，4.46(dd，J＝3.3，11.8Hz，1H)，3.00(br d，J＝13.8Hz，1H)，2.74(s，3H)，2.46-2.28(m，4H)，2.22(d，J＝7.0Hz，2H)，2.15-2.00(m，2H)，1.96(br d，J＝12.8Hz，2H)，1.24-1.08(m，2H).
Step 5: synthesis of Compound 12
Compound 10-e (210.00 mg, 348.20. Mu. Mol,1 eq) was dissolved in methanol (2 mL) and water (2 mL), and sodium hydroxide (69.64 mg,1.74mmol,5 eq) was added thereto and the mixture was stirred at 25℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product is purified by a preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile ]; acetonitrile%: 40% -70%,7 min) to obtain compound 12.
MS m/z(ESI)：575.1[M+1] + SFC retention time: 5.847 minutes; ee% = 98.5%.
1 H NMR(400MHz，CD 3 OD)δ＝7.70-7.60(m，2H)，7.54(s，1H)，6.33(s，1H)，4.99(br dd，J＝3.3，11.8Hz，1H)，4.46(dd，J＝3.3，11.8Hz，1H)，3.00(br d，J＝13.8Hz，1H)，2.74(s，3H)，2.46-2.28(m，4H)，2.22(d，J＝7.0Hz，2H)，2.15-2.00(m，2H)，1.96(br d，J＝12.8Hz，2H)，1.24-1.08(m，2H).
Example 7
The synthetic route is as follows:
step 1: synthesis of Compound 13-a
2, 6-tetramethylpiperidine (2.90 g,20.53mmol,3.48mL,1.1 eq) was added to tetrahydrofuran (50 mL), cooled to-78 ℃, n-butyllithium (2.5M, 9.70mL,1.3 eq) was added dropwise, the reaction was stirred at-78℃for 30 minutes, the temperature was raised to 0 ℃, a solution of compound 1-n in tetrahydrofuran (15 mL) was added dropwise, the reaction was stirred at 0℃for 30 minutes, and a solution of methyl iodide (3.97 g,27.99mmol,1.74mL,1.5 eq) in tetrahydrofuran (15 mL) was added dropwise. The reaction was stirred at 0deg.C for 10 min, warmed to 25deg.C and stirred for 12 hr. The temperature of the reaction mixture was lowered to 0℃and a saturated ammonium chloride solution (30 mL) was added thereto, followed by stirring for 20 minutes and extraction with ethyl acetate (20 mL. Times.2). The combined organic phases were concentrated under reduced pressure to give the crude product. Purifying the crude product with column chromatography (eluent: petroleum ether/ethyl acetate, gradient: 0-10%) to obtain compound 13-a.
1 H NMR(400MHz，CDCl 3 )δ＝1.23(s，24H)，1.07(s，3H)，0.90-0.84(m，1H).
Step 2: synthesis of Compound 13-b
2, 6-tetramethylpiperidine (2.49 g,17.63mmol,2.99mL,1.2 eq) was dissolved in tetrahydrofuran (30 mL), the system was cooled to-30deg.C and n-butyllithium (2.5M, 6.99mL,1.19 eq) was added dropwise. The reaction was stirred at-30℃for 0.5 hour, the reaction was cooled to-78℃and a solution of compound 13-a (4.14 g,14.69mmol,1 eq) in tetrahydrofuran (30 mL) was added dropwise, and stirring was carried out at-78℃for 30 minutes and compound 1-m (2.5 g,14.69mmol,2.34mL,1 eq) in tetrahydrofuran (30 mL) was added dropwise. The reaction was slowly warmed to 25℃and stirred at 25℃for 12 hours. The system was cooled to 0deg.C, saturated ammonium chloride solution (30 mL) was added dropwise, the reaction solution was stirred at 0deg.C for 20 minutes, and the filtrate was concentrated under reduced pressure after the reaction solution was filtered. Water (20 mL) and ethyl acetate (20 mL) were added for extraction. The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purifying the crude product with column chromatography (eluent: petroleum ether/ethyl acetate, gradient: 0-10%) to obtain compound 13-b.
MS m/z(ESI)：309.0[M+1] + .
1 H NMR(400MHz，CDCl 3 )δ＝4.13(q，J＝7.1Hz，2H)，3.04(br d，J＝13.6Hz，1H)，2.74(br d，J＝13.8Hz，1H)，2.56-2.45(m，1H)，2.10-1.96(m，3H)，1.90-1.81(m，1H)，1.70(s，3H)，1.62-1.55(m，2H)，1.28(s，12H)，1.26(br s，3H).
Step 3: synthesis of Compound 13-c
Compound 13-b (100 mg, 199.30. Mu. Mol,1 eq) and compound 7-c (307.14 mg, 996.51. Mu. Mol,5 eq) were dissolved in 1, 4-dioxane (3 mL), water (0.6 mL) and sodium carbonate (105.62 mg, 996.51. Mu. Mol,5 eq) were added, and 1, 1-bis (diphenylphosphine) ferrocene palladium chloride (29.17 mg, 39.86. Mu. Mol,0.2 eq) was added after nitrogen substitution. The reaction was stirred at 100℃for 12 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product. Purifying the crude product with column chromatography (eluent: petroleum ether/ethyl acetate, gradient: 0-40%) to obtain compound 13-c.
MS m/z(ESI)：603.0[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.73-7.62(m，2H)，7.43(s，1H)，5.02(dd，J＝3.5，11.8Hz，1H)，4.48(dd，J＝3.3，11.8Hz，1H)，4.18-4.11(m，2H)，2.86-2.79(m，1H)，2.76(s，3H)，2.64-2.55(m，2H)，2.44-2.28(m，2H)，2.16-2.07(m，2H)，2.04(s，3H)，2.02-1.90(m，2H)，1.69-1.60(m，1H)，1.56-1.46(m，1H)，1.28-1.25(m，3H).
Step 4: synthesis of Compound 13
Compound 13-c (30 mg, 49.74. Mu. Mol,1 eq) was dissolved in methanol (2 mL), and a solution of sodium hydroxide (1.99 mg, 49.74. Mu. Mol,1 eq) in water (2 mL) was added. The reaction was stirred at 25℃for 0.5 h. To the reaction mixture was added 2M diluted hydrochloric acid to adjust ph=2 to 3, and ethyl acetate (10 ml×3) was added for extraction. The combined organic phases were concentrated under reduced pressure. The crude product is purified by preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile;: acetonitrile%: 45% -75%,7 min) to obtain compound 13.
MS m/z(ESI)：575.0[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.73-7.60(m，2H)，7.43(s，1H)，5.02(dd，J＝3.4，11.8Hz，1H)，4.48(dd，J＝3.2，11.8Hz，1H)，2.83(br d，J＝14.5Hz，1H)，2.76(s，3H)，2.67-2.51(m，2H)，2.44-2.27(m，2H)，2.16-2.07(m，2H)，2.05(s，3H)，1.98(br t，J＝11.4Hz，2H)，1.68-1.46(m，2H).
Example 8
The synthetic route is as follows:
step 1: synthesis of Compound 14-a
Methyl triphenylphosphine bromide (23.09 g,64.63mmol,1.1 eq) was dissolved in tetrahydrofuran (160 mL), after cooling the reaction solution to-5 ℃, butyllithium (2.5M, 25.85mL,1.1 eq) was slowly added dropwise under nitrogen atmosphere, then the reaction solution was stirred at 0℃for 1 hour, then compound 1-m (10 g,58.75mmol,9.35mL,1 eq) was slowly added dropwise, and the reaction solution was stirred at 25℃for 12 hours. After the reaction, the reaction mixture was cooled to 0℃and then a saturated ammonium chloride solution (100 mL) was slowly added to the reaction mixture, followed by stirring for 10 minutes, extraction with ethyl acetate (200 mL. Times.3) was further carried out, and the organic phase was washed with saturated brine (100 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 20/1) to give compound 14-a.
1 H NMR(400MHz，CDCl 3 )δ＝4.65(s，2H)，4.13(q，J＝7.1Hz，2H)，2.51-2.25(m，3H)，2.14-1.94(m，4H)，1.65-1.54(m，2H)，1.32-1.19(m，3H).
Step 2: synthesis of Compound 14-b
Compound 14-a (3 g,17.83mmol,1 eq) was dissolved in tetrahydrofuran (150 mL), zinc-copper (20.69 g,160.49mmol,9 eq) was added, trichloroacetyl chloride (16.21 g,89.16mmol,9.95mL,5 eq) was slowly added dropwise, and the reaction was stirred at 25℃for 12 hours. The reaction solution was slowly poured into ice water, ethyl acetate (200 mL. Times.3) was added thereto to extract, and the combined organic phases were washed with saturated brine (100 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 10/1) to give compound 14-b.
1 H NMR(400MHz，CDCl 3 ) δ=4.16 (dd, j=7.2, 13.4hz, 2H), 3.04 (d, j=14.3 hz, 2H), 2.57-2.46 (m, 1H), 2.39-2.26 (m, 1H), 2.19-2.07 (m, 3H), 1.92 (br d, j=13.3 hz, 1H), 1.81-1.61 (m, 3H), 1.28 (d, j=7.0 hz, 3H),. Step 3: synthesis of Compound 14-c
Compound 14-b (2.3 g,8.24mmol,1 eq) was dissolved in glacial acetic acid (20 mL) and zinc powder (2.16 g,32.96mmol,4 eq) was added. The reaction was stirred at 50℃for 2 hours. The reaction solution was filtered, and the filter cake was quenched with 4M diluted hydrochloric acid and pH was adjusted to neutral with saturated aqueous sodium hydroxide solution. The filtrate was poured into water (50 mL), extracted with ethyl acetate (30 mL. Times.2), washed with a saturated aqueous sodium chloride solution (50 mL. Times.1) and the combined organic phases dried over anhydrous sodium sulfate, filtered and evaporated. The residue was purified by automatic column chromatography COMBI-FLASH separation (eluent: petroleum ether: ethyl acetate=100:0 to 10:1) to give compound 14-c.
1 H NMR(400MHz，CDCl 3 )δ＝4.16-1.33(m，2H)，2.77-2.76(d，J＝8.0Hz，，4H)，2.37-2.31(m，1H)，1.97-1.93(m，2H)，1.81-1.77(m，2H)，1.65-1.55(m，4H)，1.26(t，J＝7.2Hz，3H).
Step 4: synthesis of Compound 14-d
Compound 14-c (300 mg,1.43mmol,1 eq) was dissolved in tetrahydrofuran (10 mL) and lithium bis (trimethylsilyl) amide (1.0M, 1.57mL,1.1 eq) was added dropwise at-78 ℃. After completion of the dropwise addition, the mixture was stirred at-78℃for 0.5 hour, and then a mixture of N-phenylbis (trifluoromethanesulfonyl) imide (560.67 mg,1.57mmol,1.1 eq) and tetrahydrofuran (10 mL) was added dropwise. The reaction was stirred at-78℃for 0.5 hours, then warmed to-78℃and stirred for 1 hour. Water (10 mL) was added to the reaction solution, the mixture was extracted with ethyl acetate (10 mL. Times.2), and the organic phases were combined, washed with saturated aqueous sodium chloride (20 mL. Times.1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by automatic column chromatography COMBI-FLASH separation (eluent: petroleum ether: ethyl acetate=100:0 to 10:1) to give compound 14-d.
1 H NMR(400MHz，CDCl 3 )δ＝5.69(s，0.5H)，5.42(s，0.5H)，4.09-4.03(m，2H)，2.51-2.50(m，，2H)，2.26-2.19(m，1H)，1.89-1.86(m，2H)，1.59-1.51(m，6H)，1.18(t，J＝6.0Hz，，3H)，0.81-0.79((m，1H).
Step 5: synthesis of Compound 14-e
Compound 14-d (400 mg,1.17mmol,1 eq), pinacol borate (356.06 mg,1.40mmol,1.2 eq), potassium acetate (344.03 mg,3.51mmol,3 eq) and [1, 1-bis (diphenylphosphine) ferrocene ] palladium dichloride dichloromethane (95.42 mg, 116.85. Mu. Mol,0.1 eq) were added to 1, 4-dioxane (7 mL). The mixture was purged three times with nitrogen and then warmed to-78 ℃ and stirred for 3 hours. Water (10 mL) was added to the reaction solution, the mixture was extracted with ethyl acetate (10 mL), and the organic phases were combined, washed with saturated aqueous sodium chloride (10 mL. Times.1), dried over anhydrous sodium sulfate, and filtered and distilled off. The residue was purified by automatic column chromatography COMBI-FLASH separation (eluent: petroleum ether: ethyl acetate=100:0 to 10:1) to give compound 14-e.
1 H NMR(400MHz，CDCl 3 )δ＝7.15(s，0.5H)，6.86(s，0.5H)，4.08-4.02(m，2H)，2.21(d，J＝6.0Hz，，2H)，1.82-1.79(m，3H)，1.69-1.56(m，2H)，1.49-1.39(m，4H)，1.23-1.18(m，15H).
Step 6: synthesis of Compound 14-f
Compound 1-l (100 mg, 199.30. Mu. Mol,1 eq), compound 14-e (191.47 mg, 597.90. Mu. Mol,3 eq), potassium phosphate (126.91 mg, 597.90. Mu. Mol,3 eq) and [1, 1-bis (diphenylphosphino) ferrocene ] palladium dichloride dichloromethane (16.28 mg, 19.93. Mu. Mol,0.1 eq) were added to a mixture of 1, 4-dioxane (1.4 mL) and water (0.4 mL). The reaction was warmed to 80 ℃ and stirred for 3 hours. The reaction solution was poured into water (10 mL), extracted with ethyl acetate (10 ml×3), and the combined organic phases were washed with saturated aqueous sodium chloride (10 mL), dried over anhydrous sodium sulfate and filtered and distilled. The residue was purified by automatic column chromatography COMBI-FLASH separation (eluent: petroleum ether: ethyl acetate=100:0 to 2:1) to give compound 14-f.
MS m/z(ESI)：615.2[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.64-7.61(m，3H)，6.63(s，0.5H)，6.33(s，0.5H)，5.03(dd，J＝3.2Hz，1H)，4.49 (dd，J＝3.2Hz，1H)，4.16-4.11(m，2H)，2.75(s，3H)，2.53(d，J＝3.2Hz，1H)，2.39-2.32(m，3H)，1.70-1.69(m，2H)，1.69-1.49(m，7H)，1.29-1.25(m，3H).
Step 7: synthesis of Compound 14-g
Compound 14-f (90 mg, 146.32. Mu. Mol,1 eq) was dissolved in methanol (3 mL) and three nitrogen substitutions were made to add wet palladium on carbon (90 mg, 146.32. Mu. Mol,10% purity, 1.00 eq). The reaction was then replaced three times with hydrogen and stirred at 20℃under hydrogen (15 Psi) for 12 hours. The reaction solution was directly filtered, and the filtrate was concentrated under reduced pressure and dried to obtain 14-g of the compound.
MS m/z(ESI)：617.2[M+1] + .
Step 8: synthesis of Compound 14
Compound 14-g (30 mg, 48.61. Mu. Mol,1 eq) was dissolved in tetrahydrofuran (2 mL), and a mixture of sodium hydroxide (9.72 mg, 243.06. Mu. Mol,5 eq) and water (1 mL) was added. The reaction was stirred at 20℃for 1 hour. The organic solvent was directly distilled off, the pH was adjusted to 3-4 with 4N aqueous hydrochloric acid, and dichloromethane (10 ml×2) was used for extraction, and the combined organic phases were concentrated under reduced pressure and dried. The crude product is separated by preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile;: acetonitrile%: 45% -75%,7 min) to obtain compound 14.
MS m/z(ESI)：589.2[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.68-7.63(m，3H)，7.62-7.47(m，1H)，4.97-4.96(m，1H)，4.47(dd，J＝3.2Hz，1H)，3.79-3.70(m，1H)，2.75(s，3H)，2.47-2.37(m，1H)，2.32-2.29(m，1H)，2.25-2.06(m，3H)，1.93-1.89(m，4H)，1.69-1.51(m，2H)，1.49-1.44(m，4H).
Example 9
The synthetic route is as follows:
step 1: synthesis of Compound 15-a
Compound 1-h (152 g,431.59mmol,1 eq) was added to ethanol (1520 mL), the reaction cooled to 0deg.C, sodium borohydride (24.49 g,647.39mmol,1.5 eq) was slowly added, and the reaction stirred at 25deg.C for 0.5 hours. The reaction mixture was quenched slowly with saturated aqueous ammonium chloride (500 mL) and the pH was adjusted to neutral with 0.5M dilute hydrochloric acid. Most of the ethanol was removed by concentration under reduced pressure, 500mL of water was added thereto, ph=4 to 6 was adjusted with 0.5M diluted hydrochloric acid, and extraction was performed with ethyl acetate (1000 ml×2). The combined organic phases were washed successively with water (1000 mL) and saturated brine (1000 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product is pulped for 1 hour at room temperature by 500mL methyl tertiary butyl ether, filtered, and the filter cake is collected and dried under reduced pressure to obtain the compound 15-a.
MS m/z(ESI)：355.8[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.98-7.54(m，1H)，4.99(dd，J＝3.0，12.0Hz，1H)，4.41(dd，J＝2.9，12.4Hz，1H)，2.81-2.71(m，3H)，2.57-2.44(m，1H)，2.27-2.12(m，1H)，2.27-2.12(m，1H)。
Step 2: synthesis of Compound 15-b
Compound 15-a (2 g,5.61mmol,1 eq), compound 1-o (8.26 g,28.07mmol,5 eq), N, N-diisopropylethylamine (3.63 g,28.07mmol,4.89mL,5 eq) and bis (triphenylphosphine) palladium dichloride (394.08 mg, 561.46. Mu. Mol,0.1 eq) were dissolved in 1, 4-dioxane (20 mL) and water (20 mL) under nitrogen and stirred for 12 hours at 80 ℃. To the reaction solution, 1M diluted hydrochloric acid was added to adjust ph=1 to 2, water (50 mL) was further added, extraction was performed with ethyl acetate (50 ml×3), and the combined organic phases were washed with saturated brine (50 ml×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by flash column chromatography (dichloromethane/methanol=1/0 to 5/1) to give compound 15-b.
MS m/z(ESI)：444.0[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.57(dd，J＝3.3，7.5Hz，1H)，6.36(s，1H)，4.98-4.93(m，1H)，4.27(dd，J＝2.9，12.4Hz，1H)，4.14-4.07(m，1H)，2.94(br d，J＝14.3Hz，1H)，2.75(s，3H)，2.60(tt，J＝4.0，10.7Hz，1H)，2.50-2.41(m，2H)，2.31(dt，J＝3.9，12.7Hz，2H)，2.23-2.14(m，2H)，2.10-2.03(m，2H)，1.68-1.54(m，2H)，1.27-1.23(m，3H).
Step 3: synthesis of Compound 15-d
Compound 15-b (0.1 g, 225.46. Mu. Mol,1 eq) and compound 15-c (70.34 mg, 338.19. Mu. Mol,1.5 eq) were dissolved in dichloromethane (5 mL), the system was cooled to 0deg.C, and tri-N-propylcyclic phosphoric anhydride (215.21 mg, 338.19. Mu. Mol, 201.13. Mu.L, concentration: 50%,1.5 eq) and N, N-diisopropylethylamine (87.42 mg, 676.38. Mu. Mol, 117.81. Mu.L, 3 eq) were added. The reaction was stirred at 25℃for 16 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by column chromatography (eluent: petroleum ether/ethyl acetate, gradient: 0-50%). Compound 15-d is obtained.
MS m/z(ESI)：632.7[M+1] + .
Step 4: synthesis of Compound 15
Compound 15-d (50 mg, 78.92. Mu. Mol,1 eq) was dissolved in methanol (2 mL), a solution of sodium hydroxide (9.47 mg, 236.77. Mu. Mol,3 eq) in water (2 mL) was added, and the reaction was stirred at 25℃for 30 minutes. To the reaction mixture was added 2M diluted hydrochloric acid to adjust ph=3 to 4, and ethyl acetate (10 ml×3) was added for extraction. The combined organic phases were concentrated under reduced pressure. The crude product is purified by preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile;: acetonitrile%: 40% -70%,7 min) to obtain compound 15.
MS m/z(ESI)：605.1[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.78-7.68(m，2H)，7.58(s，1H)，6.38(s，1H)，5.04-4.98(m，1H)，4.47(dd，J＝3.0，11.8Hz，1H)，2.97(br d，J＝14.1Hz，1H)，2.76(s，3H)，2.59(br s，1H)，2.51-2.29(m，4H)，2.28-2.19(m，1H)，2.09(br d，J＝12.0Hz，2H)，1.71-1.55(m，2H).
Example 10
The synthetic route is as follows:
Step 1: synthesis of Compound 16-b
Compound 15-b (100 mg, 225.46. Mu. Mol,1 eq) and compound 16-a (49.75 mg, 338.19. Mu. Mol,1.5 eq) were dissolved in methylene chloride (5 mL), the reaction mixture was cooled to 0℃and then tri-N-propylcyclic phosphoric anhydride (215.21 mg, 338.19. Mu. Mol, 201.13. Mu.L, concentration: 50%,1.5 eq) and N, N-diisopropylethylamine (87.42 mg, 676.38. Mu. Mol, 117.81. Mu.L, 3 eq) were added thereto, and the reaction mixture was heated to 25℃and stirred for reaction for 12 hours. The reaction solution was concentrated directly under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 2/1) to give compound 16-b.
MS m/z(ESI)：573.2[M+1] + .
Step 2: synthesis of Compound 16
Compound 16-b (50 mg, 87.32. Mu. Mol,1 eq) was dissolved in methanol (1 mL) and water (1 mL), and sodium hydroxide (17.46 mg, 436.59. Mu. Mol,5 eq) was added thereto, and the reaction was stirred at 25℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18X 30mm X3 μm; mobile phase: [ water (0.225% formic acid) -acetonitrile ]; acetonitrile%: 35% -65%,7 min) to give compound 16.
MS m/z(ESI)：544.5[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.60-7.42(m，3H)，6.37(s，1H)，5.00(br dd，J＝3.3，11.8Hz，1H)，4.45(br dd，J＝3.3，11.8Hz，1H)，2.95(br d，J＝13.1Hz，1H)，2.74(s，3H)，2.57(brt，J＝10.8Hz，1H)，2.46(br d，J＝13.3Hz，1H)，2.41-2.26(m，3H)，2.21(br t，J＝11.4Hz，1H)，2.07(br d，J＝12.3Hz，2H)，1.69-1.55(m，2H).
Example 11
The synthetic route is as follows:
step 1: synthesis of Compound 17-b
In a round bottom flask, compound 17-a (44.25 mg, 270.55. Mu. Mol,1.2 eq), 15-b (100 mg, 225.46. Mu. Mol,1 eq), tri-n-propyl cyclic phosphoric anhydride (215.21 mg, 338.19. Mu. Mol, 201.13. Mu.L, concentration: 50%,1.5 eq) and diisopropylethylamine (87.42 mg, 676.38. Mu. Mol, 117.81. Mu.L, 3 eq) were added sequentially to dichloromethane (5 mL) and the reaction stirred under nitrogen at 25℃for 2 hours. To the reaction solution was added water (50 mL), extracted with methylene chloride (30 mL. Times.2), and the combined organic phases were washed successively with water (30 mL) and saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 10/3) to give compound 17-b.
MS m/z(ESI)：589.1[M+1] + .
Step 2: synthesis of Compound 17
Compound 17-b (60 mg, 101.85. Mu. Mol,1 eq) and sodium hydroxide (20.37 mg, 509.27. Mu. Mol,5 eq) were added to methanol (0.5 mL) and water (0.5 mL), and the reaction was stirred at 25℃for 30 minutes. The reaction solution was adjusted to ph=4 to 5 with 4M diluted hydrochloric acid, water (30 mL) was added, ethyl acetate (30 ml×2) was extracted, and the combined organic phases were washed successively with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product is purified by preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile;: 35% -65% acetonitrile, 7 min) to obtain compound 17.
MS m/z(ESI)：561.1[M+1] + .
1 H NMR(400MHz，DMSO-d 6 )δ＝12.16(br s，1H)，10.38(s，1H)，7.91(br d，J＝10.0Hz，1H)，7.68-7.59(m，1H)，7.68-7.59(m，1H)，7.41-7.27(m，1H)，6.41(s，1H)，4.92(br t，J＝10.0Hz，1H)，4.46(br d，J＝10.3Hz，1H)，2.83(br d，J＝14.1Hz，1H)，2.67(s，3H)，2.43-2.33(m，3H)，2.31-2.21(m，1H)，2.17(br d，J＝12.8Hz，2H)，1.98(br d，J＝9.0Hz，2H)，1.58-1.36(m，2H).
Example 12
The synthetic route is as follows:
step 1: synthesis of Compound 18-b
To a thumb flask was added compound 15-b (200 mg, 450.92. Mu. Mol,1 eq), compound 18-a (140.69 mg, 676.38. Mu. Mol,1.5 eq), diisopropylethylamine (174.83 mg,1.35mmol, 235.62. Mu.L, 3 eq) and tetrahydrofuran (5 mL) under nitrogen, followed by tri-n-propylcyclic phosphoric anhydride (573.90 mg, 901.84. Mu. Mol, 536.35. Mu.L, 50% purity, 2 eq). The mixture was stirred at 50℃for 12 hours. The reaction mixture was cooled to room temperature of 25℃and water (15 mL) was added thereto, pH=1 to 2 was adjusted with dilute hydrochloric acid (2M), and extraction was performed with ethyl acetate (20 mL. Times.3). The combined organic phases were washed with saturated brine (20 mL. Times.2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was isolated by automatic column chromatography COMBI-FLASH (gradient elution: petroleum ether: ethyl acetate=100:0 to 80:20) to give compound 18-b.
MS m/z(ESI)：632.8[M+1] + .
Step 2: synthesis of Compound 18
A thumb flask was charged with compound 18-b (250 mg, 394.62. Mu. Mol,1 eq), methanol (5 mL), and then a solution of sodium hydroxide (78.92 mg,1.97mmol,5 eq) in water (1 mL). The mixture was stirred at 25℃for 1 hour. Water (5 mL) was added to the reaction mixture, pH=1 to 2 was adjusted with 1M diluted hydrochloric acid, and extraction was performed with ethyl acetate (15 mL. Times.3). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated by preparative high performance liquid chromatography (column form Phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase [ water (0.225% formic acid) -acetonitrile ]; acetonitrile%: 35% -65%,7 min) to give compound 18.
MS m/z(ESI)：605.1[M+1] + .
1 H NMR(400MHz，DMSO-d 6 )δ＝12.16(br s，1H)，10.35(s，1H)，7.91(br d，J＝9.8Hz，1H)，7.72-7.61(m，2H)，7.34-7.26(m，1H)，6.41(s，1H)，4.92(brt，J＝9.9Hz，1H)，4.45(br d，J＝10.5Hz，1H)，2.83(br d，J＝14.1Hz，1H)，2.67(s，3H)，2.44-2.33(m，3H)，2.27(br d，J＝11.8Hz，1H)，2.22-2.12(m，2H)，2.04-1.92(m，2H)，1.57-1.37(m，2H).
Example 13
The synthetic route is as follows:
step 1: synthesis of Compound 19-b
Compound 15-b (100 mg, 225.46. Mu. Mol,1 eq) and compound 19-a (60.88 mg, 338.19. Mu. Mol,1.5 eq) were dissolved in methylene chloride (5 mL), the reaction mixture was cooled to 0℃and then tri-N-propylcyclic phosphoric anhydride (215.21 mg, 338.19. Mu. Mol, 201.13. Mu.L, concentration: 50%,1.5 eq) and N, N-diisopropylethylamine (87.42 mg, 676.38. Mu. Mol, 117.81. Mu.L, 3 eq) were added thereto, and the reaction mixture was heated to 25℃and stirred for reaction for 12 hours. The reaction solution was concentrated directly under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 2/1) to give compound 19-b.
MS m/z(ESI)：605.1[M+1] + .
Step 2: synthesis of Compound 19
Compound 19-b (50 mg, 82.57. Mu. Mol,1 eq) was dissolved in methanol (1 mL) and water (1 mL), and sodium hydroxide (16.51 mg, 412.86. Mu. Mol,5 eq) was added thereto, and the reaction mixture was stirred at 25℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative high performance liquid chromatography (column: phenomenexGemini-NX C18 75 x 30mm x 3 μm; mobile phase: [ water (0.225% formic acid) -acetonitrile:% 40% -70% acetonitrile, 7 min) to give compound 19.
MS m/z(ESI)：577.1[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.79(d，J＝6.0Hz，2H)，7.56(s，1H)，6.36(s，1H)，5.00(br dd，J＝3.4，11.7Hz，1H)，4.46(br dd，J＝3.1，11.9Hz，1H)，2.95(br d，J＝14.1Hz，1H)，2.74(s，3H)，2.57(br t，J＝10.8Hz，1H)，2.51-2.43(m，1H)，2.41-2.27(m，3H)，2.20(br t，J＝11.4Hz，1H)，2.12-2.04(m，2H)，1.69-1.55(m，2H).
Example 14
The synthetic route is as follows:
step 1: synthesis of Compound 20-a
Compound 15-a (10 g,28.07mmol,1 eq) was purified by SFC (column: DAICEL CHIRALPAKAD (250 mm. Times.50 mm,10 μm); mobile: [0.1% NH) 3 H 2 O，MeOH]Percent: 45% -45%) to give compound 20-a.
MS m/z(ESI)：357.7[M+1] + SFC analysis method (column: chiralpak AD-3.150 x 4.6mm I.D.,3 μm; mobile phase: carbon dioxide 40% methanol (0.05% diethylamine; flow rate: 2.5 mL/min); SFC retention time: 3.030 minutes; ee% = 99.96%.
1 H NMR(400MHz，DMSO-d 6 )δ＝7.84(s，1H)，4.76(dd，J＝2.6，11.9Hz，1H)，3.73(br dd，J＝2.3，12.3Hz，1H)，3.17(s，1H)，2.61(s，3H)，2.30(br d，J＝14.1Hz，1H)，1.95-1.86(m，1H).
Step 2: synthesis of Compound 20-b
Compound 20-a (800.00 mg,2.25mmol,1 eq), compound 10-b (2.08 g,6.74mmol,3 eq), N, N-diisopropylethylamine (1.45 g,11.23mmol,1.96mL,5 eq) and bis (triphenylphosphine) palladium dichloride (157.63 mg, 224.58. Mu. Mol,0.1 eq) were dissolved in 1, 4-dioxane (10 mL) and water (10 mL) under nitrogen, and the reaction mixture was warmed to 80℃and stirred for 4 hours. To the reaction mixture was added 1M diluted hydrochloric acid to adjust the ph=1 to 2, followed by addition of water (50 mL) and extraction with ethyl acetate (50 ml×3). The combined organic phases were washed with saturated brine (50 mL ×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by C18 reverse phase column (water/acetonitrile=1/0 to 2/1) to give compound 20-b.
MS m/z(ESI)：458.1[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.54(s，1H)，6.33(s，1H)，4.97(br dd，J＝2.9，11.9Hz，1H)，4.39(dd，J＝2.8，12.4Hz，1H)，4.16-4.10(m，2H)，2.99(br d，J＝12.0Hz，1H)，2.76(s，3H)，2.48-2.39(m，2H)，2.35-2.20(m，4H)，2.15-2.03(m，2H)，1.94(br d，J＝12.5Hz，2H)，1.28-1.24(m，3H)，1.21-1.09(m，2H).
Step 3: synthesis of Compound 20-d
Into a thumb flask, under nitrogen, 20-b (75 mg, 163.91. Mu. Mol,1 eq), 20-c (40.21 mg, 245.87. Mu. Mol,1.5 eq), diisopropylethylamine (63.55 mg, 491.74. Mu. Mol, 85.65. Mu. L,3 eq) and tetrahydrofuran (3 mL) were added, followed by tri-n-propylcyclic phosphoric anhydride (208.61 mg, 327.82. Mu. Mol, concentration: 50%,2 eq). The mixture was stirred at 50℃for 12 hours. The reaction solution was concentrated under reduced pressure. The residue was isolated by automatic column chromatography COMBI-FLASH (gradient elution: petroleum ether: ethyl acetate=100:0 to 70:30) to give compound 20-d.
MS m/z(ESI)：603.1[M+1] + .
Step 4: synthesis of Compound 20
Into a thumb flask were added compound 20-d (80 mg, 132.65. Mu. Mol,1 eq) and methanol (1 mL), followed by aqueous sodium hydroxide (2M, 331.62. Mu.L, 5 eq). The mixture was stirred at 25℃for 1 hour. Water (5 mL) was added to the reaction mixture, pH=1 to 2 was adjusted with 1M diluted hydrochloric acid, and extraction was performed with ethyl acetate (15 mL. Times.2). The combined organic phases were washed with saturated brine (30 mL) and concentrated under reduced pressure. The residue was purified by preparative high performance liquid chromatography (column form Phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase [ water (0.225% formic acid) -acetonitrile ]; acetonitrile%: 40% -70%,7 min) to give compound 20.
MS m/z(ESI)：575.0[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝8.00(t，J＝7.8Hz，1H)，7.56(s，1H)，7.37-7.24(m，1H)，6.35(s，1H)，5.03(dd，J＝3.1，11.9Hz，1H)，4.52(dd，J＝3.0，12.0Hz，1H)，3.08-2.94(m，1H)，2.80(s，3H)，2.52-2.40(m，2H)，2.38-2.28(m，2H)，2.24(d，J＝7.0Hz，2H)，2.18-2.02(m，2H)，1.98(br d，J＝12.3Hz，2H)，1.27-1.08(m，2H).
Example 15
The synthetic route is as follows:
step 1: synthesis of Compound 21-b
Compound 20-b (75.00 mg, 163.91. Mu. Mol,1 eq) and compound 21-a (40.21 mg, 245.87. Mu. Mol,1.5 eq) were dissolved in tetrahydrofuran (3 mL), the reaction solution was cooled to 0℃and then tri-N-propylcyclic phosphoric anhydride (208.61 mg, 327.82. Mu. Mol, concentration: 50%,2 eq) and N, N-diisopropylethylamine (63.55 mg, 491.74. Mu. Mol, 85.65. Mu.L, 3 eq) were added thereto, and the reaction solution was heated to 50℃and stirred for reaction for 12 hours. The reaction solution was concentrated directly under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 2/1) to give compound 21-b.
MS m/z(ESI)：603.1[M+1] + .
Step 2: synthesis of Compound 21
Compound 21-b (50 mg, 82.90. Mu. Mol,1 eq) was dissolved in methanol (1 mL) and water (1 mL), and sodium hydroxide (16.58 mg, 414.52. Mu. Mol,5 eq) was added thereto, and the reaction mixture was stirred at 25℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative high performance liquid chromatography (column: phenomenexGemini-NX C18 75 x 30mm x 3 μm; mobile phase: [ water (0.225% formic acid) -acetonitrile:% 40% -70% acetonitrile, 7 min) to give compound 21.
MS m/z(ESI)：575.1[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.68(dt，J＝5.6，8.7Hz，1H)，7.55(s，1H)，7.15(dt，J＝2.0，8.9Hz，1H)，6.33(s，1H)，5.02(dd，J＝3.1，12.2Hz，1H)，4.51(dd，J＝3.1，11.9Hz，1H)，3.00(br d，J＝13.8Hz，1H)，2.80(s，3H)，2.49-2.39(m，2H)，2.37-2.26(m，2H)，2.22(d，J＝7.0Hz，2H)，2.16-2.00(m，2H)，1.97(br d，J＝11.5Hz，2H)，1.25-1.09(m，2H).
Example 16
The synthetic route is as follows:
step 1: synthesis of Compound 22-b
Compound 20-b (75 mg, 163.91. Mu. Mol,1 eq) and compound 22-a (40.21 mg, 245.87. Mu. Mol,1.5 eq) were dissolved in tetrahydrofuran (3 mL) under nitrogen, the reaction mixture was cooled to 0℃and then tri-N-propylcyclic phosphoric anhydride (208.61 mg, 327.82. Mu. Mol, 194.97. Mu.L, concentration: 50%,2 eq) and N, N-diisopropylethylamine (63.55 mg, 491.74. Mu. Mol, 85.65. Mu.L, 3 eq) were added thereto, and the reaction mixture was heated to 50℃and stirred for 12 hours. The reaction solution was concentrated directly under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 2/1) to give compound 22-b.
MS m/z(ESI)：603.1[M+1] + .
Step 2: synthesis of Compound 22
Compound 22-b (10 mg, 16.58. Mu. Mol,1 eq) was dissolved in methanol (1 mL) and water (1 mL), and sodium hydroxide (3.32 mg, 82.90. Mu. Mol,5 eq) was added thereto and the mixture was stirred at 25℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product is purified by a preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile: acetonitrile%: 40% -70%,7 min) to obtain compound 22.
MS m/z(ESI)：575.1[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.84(dd，J＝8.0，11.8Hz，1H)，7.58-7.50(m，2H)，6.33(s，1H)，5.02(dd，J＝3.0，12.0Hz，1H)，4.52(dd，J＝3.3，12.0Hz，1H)，3.00(br d，J＝14.1Hz，1H)，2.82(s，3H)，2.54-2.40(m，2H)，2.37-2.27(m，2H)，2.23(d，J＝7.0Hz，2H)，2.16-2.00(m，2H)，1.97(br d，J＝11.8Hz，2H)，1.23-1.09(m，2H).
Example 17
The synthetic route is as follows:
step 1: synthesis of Compound 23-b
23-a (500 mg,3.12mmol,1 eq) and wet palladium on carbon (67.47 mg, 31.23. Mu. Mol,5% purity, 0.01 eq) were added to methanol (50 mL) in a round bottom flask under nitrogen, then replaced three times with hydrogen and the reaction stirred at 40℃for 1 hour under H2 (25 psi) atmosphere. The reaction solution was filtered and concentrated under reduced pressure to give compound 23-b.
MS m/z(ESI)：130.8[M+1] + .
1 H NMR(400MHz，DMSO-d 6 ) δ=7.28 (t, j=2.6 hz, 1H), 7.06 (ddd, j= 2.4,8.6, 11.9hz, 1H), 5.54 (br s, 2H) ·step 2: synthesis of Compound 23-c
20-b (50 mg, 109.27. Mu. Mol,1 eq), 23-b (17.06 mg, 131.13. Mu. Mol,1.2 eq), tri-n-propyl cyclic phosphoric anhydride (139.08 mg, 218.55. Mu. Mol, concentration: 50%,2 eq) and diisopropylethylamine (42.37 mg, 327.82. Mu. Mol, 57.10. Mu.L, 3 eq) were added to tetrahydrofuran (5 mL), and the reaction solution was stirred at 50℃for 0.5 hours under nitrogen. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL. Times.2). The combined organic phases were washed successively with water (30 mL) and saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 10/3) to give compound 23-c.
MS m/z(ESI)：570.0[M+1] + .
Step 3: synthesis of Compound 23
Compound 23-c (40 mg, 70.22. Mu. Mol,1 eq) and sodium hydroxide (28.09 mg, 702.19. Mu. Mol,10 eq) were added to methanol (1 mL) and water (1 mL), and the reaction solution was stirred at 25℃for 1 hour. The reaction mixture was adjusted to ph=4 to 5 with 4M diluted hydrochloric acid, water (30 mL) was added thereto, and extraction was performed with ethyl acetate (30 ml×2). The combined organic phases were washed successively with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product is purified by a preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile: acetonitrile%: 35% -55%,7 min) to obtain compound 23.
MS m/z(ESI)：541.9[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝8.29(ddd，J＝2.3，8.7，11.2Hz，1H)，8.21(t，J＝2.0Hz，1H)，7.57(s，1H)，6.35(s，1H)，5.02(br dd，J＝3.0，11.8Hz，1H)，4.54(dd，J＝3.1，11.7Hz，1H)，3.09-2.94(m，1H)，2.77(s，3H)，2.49-2.20(m，6H)，2.19-1.92(m，4H)，1.29-1.04(m，2H).
Example 18
The synthetic route is as follows:
step 1: synthesis of Compound 24-b
To a thumb flask, compound 20-b (75 mg, 163.91. Mu. Mol,1 eq), compound 24-a (31.61 mg, 245.87. Mu. Mol,1.5 eq), diisopropylethylamine (63.55 mg, 491.73. Mu. Mol, 85.65. Mu. L,3 eq) and tetrahydrofuran (3 mL) were added under nitrogen, followed by tri-n-propylcyclic phosphoric anhydride (208.61 mg, 327.82. Mu. Mol, 194.97. Mu.L, concentration: 50%,2 eq). The mixture was stirred at 50℃for 12 hours. The reaction solution was concentrated under reduced pressure. The residue was isolated by automatic column chromatography COMBI-FLASH (gradient elution: petroleum ether: ethyl acetate=100:0 to 50:50) to give compound 24-b.
MS m/z(ESI)：568.0[M+1] + .
Step 2: synthesis of Compound 24
In a thumb flask was added compound 24-b (40 mg, 70.41. Mu. Mol,1 eq) and methanol (1 mL), followed by sodium hydroxide (2M, 0.5mL,14.20 eq). The mixture was stirred at 25℃for 1 hour. Water (5 mL) was added to the reaction mixture, pH=5 to 6 was adjusted with 1M diluted hydrochloric acid, and extraction was performed with ethyl acetate (15 mL. Times.3). The combined organic phases were washed with saturated brine (30 mL) and concentrated under reduced pressure. The residue was purified by preparative high performance liquid chromatography (column form Phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: [ water (0.225% formic acid) -acetonitrile:% 40% -70%,7 min) to give compound 24.
MS m/z(ESI)：540.1[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝8.72(br s，1H)，8.41-8.23(m，2H)，7.57(s，1H)，6.35(s，1H)，5.02(br d，J＝8.8Hz，1H)，4.56(dd，J＝3.0，11.8Hz，1H)，3.11-2.94(m，1H)，2.78(s，3H)，2.50-2.38(m，2H)，2.37-2.28(m，2H)，2.25(d，J＝7.0Hz，2H)，2.17-1.95(m，4H)，1.27-1.06(m，2H).
Example 19
The synthetic route is as follows:
step 1: synthesis of Compound 25-b
To a thumb flask, under nitrogen, was added compound 20-b (50 mg, 109.27. Mu. Mol,1 eq), compound 25-a (21.07 mg, 163.91. Mu. Mol,1.5 eq) and dichloromethane (1.5 mL), followed by pyridine (43.22 mg, 546.37. Mu. Mol, 44.10. Mu.L, 5 eq) and phosphorus oxychloride (20.11 mg, 131.13. Mu. Mol, 12.19. Mu.L, 1.2 eq) and the mixture was stirred at 20℃for 1 hour. The reaction solution was concentrated under reduced pressure to give crude compound 25-b.
MS m/z(ESI)：568.1[M+1] + .
Step 2: synthesis of Compound 25
In a thumb flask were added compound 25-b (100 mg, 176.02. Mu. Mol,1 eq) and methanol (2 mL), followed by aqueous sodium hydroxide (2M, 1mL,11.36 eq). The mixture was stirred at 25℃for 1 hour. To the reaction mixture was added water (5 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (15 ml×3). The combined organic phases were washed with saturated brine (30 mL) and concentrated under reduced pressure. The residue was separated by preparative high performance liquid chromatography (column form: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: [ water (0.225% formic acid) -acetonitrile:% 30% -60%,7 min) to give compound 25.
MS m/z(ESI)：540.2[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝8.26(d，J＝5.5Hz，1H)，7.89(d，J＝1.8Hz，1H)，7.63(dd，J＝1.8，5.8Hz，1H)，7.57(s，1H)，6.35(s，1H)，5.01(dd，J＝3.8，11.5Hz，1H)，4.58(dd，J＝3.5，11.5Hz，1H)，3.02(br d，J＝13.6Hz，1H)，2.75(s，3H)，2.47-2.29(m，4H)，2.25(d，J＝6.8Hz，2H)，2.17-2.02(m，2H)，1.98(br d，J＝12.0Hz，2H)，1.31-1.02(m，2H).
Example 20
The synthetic route is as follows:
step 1: synthesis of Compound 26-b
Compound 20-b (50 mg, 109.27. Mu. Mol,1 eq) and compound 26-a (22.31 mg, 163.91. Mu. Mol,1.5 eq) were dissolved in tetrahydrofuran (2 mL) under nitrogen, the reaction mixture was cooled to 0℃and then tri-N-propylcyclic phosphoric anhydride (139.08 mg, 218.55. Mu. Mol, 129.98. Mu.L, 50% purity, 2 eq) and N, N-diisopropylethylamine (42.37 mg, 327.82. Mu. Mol, 57.10. Mu.L, 3 eq) were slowly added thereto, and the reaction mixture was heated to 50℃and stirred for 12 hours. The reaction solution was concentrated directly under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 2/1) to give compound 26-b.
MS m/z(ESI)：576.1[M+1] + .
Step 2: synthesis of Compound 26
Compound 26-b (20 mg, 34.74. Mu. Mol,1 eq) was dissolved in methanol (1 mL) and water (1 mL), and sodium hydroxide (6.95 mg, 173.71. Mu. Mol,5 eq) was added thereto, and the reaction mixture was stirred at 25℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product is separated by a preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile: acetonitrile%: 35% -65%,7 min) to obtain compound 26.
MS m/z(ESI)：548.2[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.89-7.83(m，2H)，7.55(s，1H)，7.34-7.28(m，1H)，6.33(s，1H)，5.00(dd，J＝3.6，11.7Hz，1H)，4.52(dd，J＝3.4，11.7Hz，1H)，3.00(br d，J＝14.6Hz，1H)，2.75(s，3H)，2.44-2.36(m，2H)，2.34-2.25(m，2H)，2.22(d，J＝7.0Hz，2H)，2.15-2.03(m，2H)，1.97(br d，J＝12.5Hz，2H)，1.21-1.10(m，2H).
Example 21
The synthetic route is as follows:
step 1: synthesis of Compound 27-b
In a round bottom flask, compound 20-b (60 mg, 131.13. Mu. Mol,1 eq), 27-a (24.01 mg, 157.36. Mu. Mol,1.2 eq), tri-n-propyl cyclic phosphoric anhydride (166.89 mg, 262.26. Mu. Mol, purity: 50%,2 eq) and diisopropylethylamine (50.84 mg, 393.39. Mu. Mol, 68.52. Mu.L, 3 eq) were added to tetrahydrofuran (6 mL) and the reaction stirred at 50℃for 0.5 h. To the reaction solution was added water (50 mL), extracted with ethyl acetate (30 mL. Times.2), and the organic phases were combined, washed with water (30 mL) and saturated brine (30 mL) in this order, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 10/3) to give compound 27-b.
MS m/z(ESI)：591.9[M+1] + .
Step 2: synthesis of Compound 27
Compound 27-b (40 mg, 67.55. Mu. Mol,1 eq) and sodium hydroxide (27.02 mg, 675.53. Mu. Mol,10 eq) were added to methanol (1 mL) and water (1 mL), and the reaction was stirred at 25℃for 1 hour. The reaction mixture was adjusted to ph=4 to 5 with 4M hydrochloric acid, water (30 mL) was added, and extraction was performed with ethyl acetate (30 ml×2). The combined organic phases were washed successively with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product is purified by preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile: acetonitrile%: 45% -65%,7 min) to obtain compound 27.
MS m/z(ESI)：564.0[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝8.18-7.87(m，2H)，7.69-7.40(m，2H)，6.35(s，1H)，5.02(dd，J＝3.4，11.7Hz，1H)，4.54(dd，J＝3.4，11.7Hz，1H)，3.15-2.90(m，1H)，2.77(s，3H)，2.55-2.20(m，6H)，2.15-1.88(m，4H)，1.26-1.06(m，2H).
Example 22
The synthetic route is as follows:
step 1: synthesis of Compound 28-b
Compound 28-a (500.00 mg,2.37mmol,1 eq) was dissolved in t-butanol (10 mL), diphenyl azide phosphate (784.28 mg,2.85mmol, 617.54. Mu.L, 1.2 eq) and triethylamine (288.37 mg,2.85mmol, 396.66. Mu.L, 1.2 eq) were added, and the reaction mixture was heated to 90℃and stirred for 6 hours. Water (15 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (15 mL. Times.3). The combined organic phases were washed with saturated brine (15 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 2/1) to give compound 28-b.
1 H NMR(400MHz，CDCl 3 )δ＝8.04(br d，J＝11.5Hz，1H)，6.73-6.56(m，1H)，1.54(s，9H)
Step 2: synthesis of Compound 28-c
Compound 28-b (400.00 mg,1.42mmol,1 eq) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (2 mL) was added, and the reaction was stirred at 25℃for 0.5 h. The reaction solution was concentrated under reduced pressure to give crude compound 28-c.
1 H NMR(400MHz，CDCl 3 )δ＝6.54(td，J＝7.6，11.2Hz，1H)，3.75(br s，2H).
Step 3: synthesis of Compound 28-d
Compound 20-b (50 mg, 109.27. Mu. Mol,1 eq) and compound 28-c (29.76 mg, 163.9L. Mu. Mol,1.5 eq) were dissolved in tetrahydrofuran (2 mL) under nitrogen, the reaction solution was cooled to 0℃and then tri-N-propylcyclic phosphoric anhydride (139.08 mg, 218.54. Mu. Mol,50% purity, 2 eq) and N, N-diisopropylethylamine (42.37 mg, 327.81. Mu. Mol, 57.10. Mu.L, 3 eq) were slowly added thereto, and the reaction solution was heated to 50℃and stirred for 12 hours. The reaction solution was concentrated directly under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 2/1) to give compound 28-d.
MS m/z(ESI)：621.1[M+1] + .
Step 4: synthesis of Compound 28
Compound 28-d (20.00 mg, 32.20. Mu. Mol,1 eq) was dissolved in methanol (1 mL) and water (1 mL), and sodium hydroxide (6.44 mg, 161.01. Mu. Mol,5 eq) was added thereto, and the reaction mixture was stirred at 25℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product is separated by a preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile: acetonitrile%: 40% -70%,7 min) to obtain compound 28.
MS m/z(ESI)：593.1[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.92-7.85(m，1H)，7.55(s，1H)，6.33(s，1H)，5.01(br d，J＝8.5Hz，1H)，4.55-4.50(m，1H)，3.00(br d，J＝13.3Hz，1H)，2.78(s，3H)，2.45-2.39(m，2H)，2.35-2.26(m，2H)，2.22(d，J＝7.0Hz，2H)，2.15-2.03(m，2H)，1.96(br d，J＝12.0Hz，2H)，1.15(br d，J＝12.5Hz，2H).
Example 23
The synthetic route is as follows:
step 1: synthesis of Compound 29-a
In a round bottom flask, 20-b (70 mg, 152.98. Mu. Mol,1 eq), 3-chloro-4-fluoroaniline (26.72 mg, 183.58. Mu. Mol,1.2 eq), tri-n-propylcyclic phosphoric anhydride (194.71 mg, 305.97. Mu. Mol, 181.97. Mu.L, concentration: 50%,2 eq) and diisopropylethylamine (79.09 mg, 611.94. Mu. Mol, 106.59. Mu.L, 4 eq) were added to tetrahydrofuran (5 mL), and the reaction mixture was stirred under nitrogen at 50℃for 1 hour. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL. Times.2). The combined organic phases were washed successively with water (30 mL) and saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 10/3) to give compound 29-a.
MS m/z(ESI)：585.1[M+1] + .
Step 2: synthesis of Compound 29
Compound 29-a (50 mg, 85.45. Mu. Mol,1 eq) and sodium hydroxide (17.09 mg, 427.27. Mu. Mol,5 eq) were added to methanol (0.5 mL) and water (0.5 mL), and the reaction was stirred at 25℃for 60 min. The reaction mixture was adjusted to ph=4 to 5 with 4M diluted hydrochloric acid, water (30 mL) was added thereto, and extraction was performed with ethyl acetate (30 ml×2). The combined organic phases were washed successively with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by preparative high performance liquid chromatography (column form: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: [ water (0.225% formic acid) -acetonitrile: [ acetonitrile%: 40% -70%,7 min.) to give compound 29.
MS m/z(ESI)：556.8[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.91(dd，J＝2.6，6.7Hz，1H)，7.58-7.49(m，2H)，7.23(t，J＝8.9Hz，1H)，6.35(s，1H)，5.02(dd，J＝3.3，11.8Hz，1H)，4.44(dd，J＝3.3，11.8Hz，1H)，3.11-2.91(m，1H)，2.77(s，3H)，2.51-2.19(m，6H)，2.19-1.92(m，4H)，1.29-1.01(m，2H).
Step 3: synthesis of Compounds 30 and 31
Compound 29 (20 mg, 35.90. Mu. Mol) was purified by SFC (column: DAICEL CH)IRALCEL OJ (250 mm x 30mm,10 μm); mobile phase: [0.1% NH 3 H 2 O，MeOH]Percent: 40% -40%) to give compounds 30 and 31.
SFC analysis method: column type: chiralCel OJ-H150X 4.6mm I.D.,5 μm; mobile phase: methanol (0.05% diethylamine); flow rate: 2.5mL/min.
Compound 30: MS m/z (ESI): 556.9[ M+1 ]] + SFC retention time: 7.427 minutes.
1H NMR(400MHz，CD 3 OD)δ＝7.91(dd，J＝2.6，6.7Hz，1H)，7.68-7.44(m，2H)，7.24(t，J＝8.9Hz，1H)，6.35(s，1H)，5.02(dd，J＝3.4，11.7Hz，1H)，4.44(dd，J＝3.3，11.8Hz，1H)，3.07-2.93(m，1H)，2.77(s，3H)，2.51-2.26(m，4H)，2.22(d，J＝7.0Hz，2H)，2.16-1.93(m，4H)，1.28-1.08(m，2H).
Compound 31: MS m/z (ESI): 556.9[ M+1 ] ] + SFC retention time: 7.808 minutes.
1H NMR(400MHz，CD 3 OD)δ＝7.91(dd，J＝2.6，6.7Hz，1H)，7.69-7.47(m，2H)，7.24(t，J＝9.0Hz，1H)，6.35(s，1H)，5.10-4.95(m，1H)，4.44(dd，J＝3.0，11.5Hz，1H)，3.02(br d，J＝14.1Hz，1H)，2.77(s，3H)，2.52-2.26(m，4H)，2.22(br d，J＝6.8Hz，2H)，2.17-1.93(m，4H)，1.27-1.08(m，2H).
Example 24
The synthetic route is as follows:
step 1: synthesis of Compound 32-a
Compound 20-b (50 mg, 109.27. Mu. Mol,1 eq) and compound 15-c (22.73 mg, 109.27. Mu. Mol,1 eq) were dissolved in tetrahydrofuran (2 mL) under nitrogen, the reaction mixture was cooled to 0℃and then tri-N-propylcyclic phosphoric anhydride (139.08 mg, 218.55. Mu. Mol, 129.98. Mu.L, concentration: 50%,2 eq) and N, N-diisopropylethylamine (42.37 mg, 327.82. Mu. Mol, 57.10. Mu.L, 3 eq) were slowly added thereto, and the mixture was stirred for reaction for 12 hours while heating to 50 ℃. The reaction solution was concentrated directly under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 2/1) to give compound 32-a.
MS m/z(ESI)：647.0[M+1] + .
Step 2: synthesis of Compound 32
Compound 32-a (40 mg, 61.77. Mu. Mol,1 eq) was dissolved in methanol (1 mL) and water (1 mL), and sodium hydroxide (12.35 mg, 308.86. Mu. Mol,5 eq) was added thereto, and the reaction mixture was stirred at 25℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product is separated by a preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile ]; acetonitrile%: 40% -70%,7 min) to obtain compound 32.
MS m/z(ESI)：618.9[M+1] + .
Step 3: synthesis of Compounds 33 and 34
Compound 32 (25 mg, 40.36. Mu. Mol,1 eq) was purified by SFC (column: DAICEL CHIRALPAK AS (250 mm. Times.30 mm,10 μm); mobile phase: [0.1% NH) 3 H 2 O，EtOH]Percent: 35% -35%) to give compounds 33 and 34.
SFC analysis method: column type: chiralPak AS-3X 4.6mm I.D.,3 μm; mobile phase: isopropanol (0.05% diethylamine); flow rate: 2.5mL/min.
Compound 33: MS m/z (ESI): 619.0[ M+1 ]] + SFC retention time: 5.872 minutes, ee% = 92.5%.
1 H NMR(400MHz，CD 3 OD)δ＝7.80-7.66(m，2H)，7.55(s，1H)，6.33(s，1H)，4.99(dd，J＝3.4，11.7Hz，1H)，4.46(dd，J＝3.5，11.8Hz，1H)，3.00(br d，J＝13.1Hz，1H)，2.74(s，3H)，2.45-2.27(m，4H)，2.26-2.18(m，2H)，2.15-2.00(m，2H)，1.97(br d，J＝11.5Hz，2H)，1.22-1.10(m，2H).
Compound 34: MS m/z (ESI): 619.0[ M+1 ]] + SFC analysis method (column: chiralpak AD-3.150 x 4.6mm I.D.,3 μm; mobile phase: isopropanol (0.05% diethylamine; flow rate: 2.5 mL/min), retention time: 6.150 min, ee% = 84.8%.
1 H NMR(400MHz，CD 3 OD)δ＝7.79-7.65(m，2H)，7.55(s，1H)，6.33(s，1H)，4.99(dd，J＝3.4，11.7Hz，1H)，4.46(dd，J＝3.5，11.8Hz，1H)，3.00(br d，J＝13.8Hz，1H)，2.74(s，3H)，2.45-2.27(m，4H)，2.22(d，J＝6.8Hz，2H)，2.15-2.00(m，2H)，1.99-1.93(m，2H)，1.21-1.09(m，2H).
Example 25
The synthetic route is as follows:
step 1: synthesis of Compound 35-a
Compound 20-b (325 mg, 710.28. Mu. Mol,1 eq) and compound 17-a (174.25 mg,1.07mmol,1.5 eq) were dissolved in methylene chloride (15 mL) under nitrogen, and after cooling the reaction solution to 0℃phosphorus oxychloride (163.36 mg,1.07mmol, 99.01. Mu.L, 1.5 eq) and pyridine (280.92 mg,3.55mmol, 286.65. Mu.L, 5 eq) were slowly added thereto, and the reaction solution was stirred at 0℃for 1 hour. The reaction solution was concentrated directly under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 2/1) to give compound 35-a.
MS m/z(ESI)：603.1[M+1] + .
Step 2: synthesis of Compounds 35-b and 35-c
Compound 35-a (220 mg, 364.78. Mu. Mol,1 eq) was purified by SFC (column: DAICEL CHIRALCEL OJ (250 mm. Times.30 mm,10 μm); mobile phase: [0.1% NH) 3 H 2 O，EtOH]Isolation of%40% -40%) gives compounds 35-b and 35-c.
Compound 35-b: SFC analysis method (column: chiralCel OJ-H150X 4.6mm I.D.,5um; mobile phase: ethanol (0.05% diethylamine; flow rate: 2.5 mL/min); MS m/z (ESI): 603.1[ M+1 ]] + SFC retention time: 5.635 minutes, ee% = 100%.
Compound 35-b: MS m/z (ESI): 603.1[ M+1 ]] + SFC retention time: for 6.192 min, ee% = 98.8%.
Step 3: synthesis of Compound 35
Compound 35-b (90 mg, 149.23. Mu. Mol,1 eq) was dissolved in methanol (3 mL) and water (3 mL), and sodium hydroxide (29.84 mg, 746.14. Mu. Mol,5 eq) was added thereto, and the reaction mixture was stirred at 25℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative high performance liquid chromatography (column form Phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase [ water (0.225% formic acid) -acetonitrile ]; acetonitrile%: 40% -70%,7 min) to give compound 35.
SFC analysis method: column type: chiralCel OJ-H150X 4.6mm I.D.,5 μm; mobile phase: ethanol (0.05% diethylamine); flow rate: MS m/z (ESI) at 2.5 mL/min: 575.1[ M+1 ]] + SFC retention time: 5.950 min, ee% = 98.3%.
1 H NMR(400MHz，CD 3 OD)δ＝7.68(dt，J＝5.8，8.7Hz，1H)，7.55(s，1H)，7.15(dt，J＝2.3，8.9Hz，1H)，6.34(s，1H)，5.02(dd，J＝3.1，12.2Hz，1H)，4.51(dd，J＝3.0，12.0Hz，1H)，3.00(br d，J＝13.6Hz，1H)，2.80(s，3H)，2.47-2.40(m，2H)，2.37-2.27(m，2H)，2.22(d，J＝7.0Hz，2H)，2.16-2.01(m，2H)，1.97(br d，J＝12.3Hz，2H)，1.23-1.09(m，2H).
Step 4: synthesis of Compound 36
Into a thumb flask were added compound 35-c (100 mg, 165.81. Mu. Mol,1 eq) and methanol (2 mL), followed by aqueous sodium hydroxide (2M, 1mL,12.06 eq) and stirring at 25℃for 30 min. The reaction mixture was adjusted to ph=1 to 2 with 1M diluted hydrochloric acid and extracted with ethyl acetate (15 ml×3). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated by preparative high performance liquid chromatography (column form: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: [ water (0.225% formic acid) -acetonitrile ]; acetonitrile%: 35% -65%,7 min). Compound 36 was obtained.
MS m/z(ESI)：575.1[M+1] + SFC retention time: 6.251 minutes, ee% = 100%.
1 H NMR(400MHz，CD 3 OD)δ＝7.74-7.65(m，1H)，7.57(s，1H)，7.17(dt，J＝2.0，8.9Hz，1H)，6.35(s，1H)，5.04(dd，J＝3.3，12.0Hz，1H)，4.53(dd，J＝3.0，12.0Hz，1H)，3.02(br d，J＝14.8Hz，1H)，2.82(s，3H)，2.51-2.40(m，2H)，2.39-2.29(m，2H)，2.24(d，J＝6.8Hz，2H)，2.18-1.94(m，4H)，1.28-1.04(m，2H).
Example 26
The synthetic route is as follows:
step 1: synthesis of Compound 37-a
Compound 20-b (45 mg, 98.35. Mu. Mol,1 eq) and compound 18-a (30.68 mg, 147.52. Mu. Mol,1.5 eq) were dissolved in tetrahydrofuran (2 mL) under nitrogen, the reaction mixture was cooled to 0℃and then tri-N-propylcyclic phosphoric anhydride (125.17 mg, 196.69. Mu. Mol, 116.98. Mu.L, concentration: 50%,2 eq) and N, N-diisopropylethylamine (38.13 mg, 295.04. Mu. Mol, 51.39. Mu.L, 3 eq) were added thereto, and the reaction mixture was stirred at 50℃for 12 hours. The reaction solution was concentrated directly under reduced pressure. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1/0 to 2/1) to give compound 37-a.
MS m/z(ESI)：647.1[M+1] + .
Step 2: synthesis of Compound 37
Compound 37-a (20 mg, 30.89. Mu. Mol,1 eq) was dissolved in methanol (1 mL) and water (1 mL), and sodium hydroxide (6.18 mg, 154.43. Mu. Mol,5 eq) was added thereto, and the reaction mixture was stirred at 25℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative high performance liquid chromatography (column form Phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase [ water (0.225% formic acid) -acetonitrile ]; acetonitrile%: 40% -70%,7 min) to give compound 37.
MS m/z(ESI)：618.8[M+1] + .
1 H NMR(400MHz，DMSO-d6)δ＝7.71-7.60(m，2H)，7.37-7.23(m，1H)，6.37(s，1H)，4.91(br d，J＝10.0Hz，1H)，4.44(dd，J＝2.5，12.0Hz，1H)，2.87(br d，J＝14.1Hz，1H)，2.66(s，3H)，2.42-2.33(m，2H)，2.26-2.12(m，4H)，2.09-1.84(m，4H)，1.17-1.05(m，1H).
Example 27
The synthetic route is as follows:
step 1: synthesis of Compound 38-a
Compound 20-a (0.3 g, 842.18. Mu. Mol,1 eq), compound 1-o (371.65 mg,1.26mmol,1.5 eq), N, N-diisopropylethylamine (544.22 mg,4.21mmol, 733.45. Mu.L, 5 eq) and dichlorobis (triphenylphosphine) palladium (59.11 mg, 84.22. Mu. Mol,0.1 eq) were dissolved in 1, 4-dioxane (4 mL) and water (4 mL) under nitrogen and the mixture was stirred at 80℃for 12 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound 38-a, the crude product was used directly in the next reaction.
MS m/z(ESI)：444.1[M+1] + .
Step 2: synthesis of Compound 38-b
Compound 38-a (100 mg, 225.46. Mu. Mol,1 eq) and compound 17-a (55.31 mg, 338.19. Mu. Mol,1.5 eq) were dissolved in methylene chloride (3 mL), the reaction solution was cooled to 0℃and phosphorus oxychloride (69.14 mg, 450.92. Mu. Mol, 41.90. Mu.L, 2 eq) and pyridine (89.17 mg,1.13mmol, 90.99. Mu.L, 5 eq) were slowly added and stirred at 0℃for 1 hour. The reaction solution was concentrated under reduced pressure to give crude compound 38-b.
MS m/z(ESI)：589.1[M+1] + .
Step 3: synthesis of Compound 38
Compound 38-b (30 mg, 50.93. Mu. Mol,1 eq) was dissolved in methanol (1 mL) and water (1 mL), and sodium hydroxide (10.19 mg, 254.65. Mu. Mol,5 eq) was added thereto and the mixture was stirred at 25℃for 2 hours. To the reaction mixture was added water (10 mL), which was then adjusted to ph=1-2 with 1M diluted hydrochloric acid, followed by extraction with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product is separated by a preparative high performance liquid chromatography (column type: phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase: water (0.225% formic acid) -acetonitrile ]; acetonitrile%: 40% -70%,7 min) to obtain compound 38.
MS m/z(ESI)：561.1[M+1] + .
1H NMR(400MHz，CD 3 OD)δ＝7.68(ddd，J＝5.6，8.2，9.2Hz，1H)，7.57(s，1H)，7.15(dt，J＝2.0，8.9Hz，1H)，6.37(s，1H)，5.02(dd，J＝3.0，12.0Hz，1H)，4.51(dd，J＝3.0，12.0Hz，1H)，2.96(br d，J＝13.8Hz，1H)，2.83-2.76(m，3H)，2.63-2.52(m，1H)，2.45(td，J＝3.4，13.9Hz，2H)，2.37-2.17(m，3H)，2.12-2.04(m，2H)，1.70-1.54(m，2H).
Example 28
The synthetic route is as follows:
step 1: synthesis of Compound 39-a
The compound 20-b (50 mg, 109.27. Mu. Mol,1 eq), 16-a (24.11 mg, 163.91. Mu. Mol, 17.35. Mu. L,1.5 eq) and methylene chloride (1.5 mL) were added to a thumb flask under nitrogen, and then pyridine (43.22 mg, 546.37. Mu. Mol, 44.10. Mu.L, 5 eq) and phosphorus oxychloride (20.11 mg, 131.13. Mu. Mol, 12.19. Mu.L, 1.2 eq) were added, and the mixture was stirred at 0℃for 1 hour. The reaction solution was concentrated under reduced pressure to give crude compound 39-a.
MS m/z(ESI)：587.1[M+1] + .
Step 2: synthesis of Compound 39
In a thumb flask were added compound 39-a (60 mg, 102.28. Mu. Mol,1 eq) and methanol (1 mL), followed by sodium hydroxide (2M, 0.5mL,9.78 eq) and then stirred at 25℃for 1 hour. The reaction mixture was adjusted to ph=1 to 2 with 1M diluted hydrochloric acid and extracted with ethyl acetate (15 ml×3). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated by preparative high performance liquid chromatography (column form Phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase [ water (0.225% formic acid) -acetonitrile ]; acetonitrile%: 35% -65%,7 min) to give compound 39.
MS m/z(ESI)：559.1[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.56(s，1H)，7.54-7.47(m，1H)，7.22-7.09(m，1H)，6.35(s，1H)，5.03(dd，J＝3.1，11.9Hz，1H)，4.53(dd，J＝3.0，12.0Hz，1H)，3.02(br d，J＝14.1Hz，1H)，2.81(s，3H)，2.51-2.40(m，2H)，2.38-2.28(m，2H)，2.24(d，J＝7.0Hz，2H)，2.17-1.95(m，4H)，1.29-1.07(m，2H).
Step 3: synthesis of Compounds 40 and 41
Compound 39 (25 mg, 44.76. Mu. Mol,1 eq) was purified by chiral SFC (column: DAICEL CHIRALCEL OJ-H (250 mm. Times.30 mm,5 μm); mobile phase: [0.1% NH) 3 H 2 O，MeOH]Percent: 40% -40%) to give compounds 40 and 41.
SFC analysis method: column type: chiralcel OJ-3.150 x 4.6mm I.D.,3 μm; mobile phase: methanol (0.05% diethylamine); flow rate: 2.5mL/min.
Compound 40: MS m/z (ESI): 558.9[ M+1 ]] + SFC retention time: 1.864 minutes, ee% = 96.2%.
1 H NMR(400MHz，CD 3 OD)δ＝7.57(s，1H)，7.54-7.47(m，1H)，7.21-7.12(m，1H)，6.35(s，1H)，5.04(dd，J＝3.1，11.9Hz，1H)，4.53(dd，J＝3.0，12.0Hz，1H)，3.02(br d，J＝13.8Hz，1H)，2.81(s，3H)，2.49-2.41(m，2H)，2.39-2.27(m，2H)，2.24(d，J＝7.0Hz，2H)，2.18-2.03(m，2H)，1.99(br d，J＝12.8Hz，2H)，1.27-1.07(m，2H).
Compound 41: MS m/z (ESI): 558.9[ M+1 ] ] + SFC retention time: 2.307 minutes, ee% = 80.5%.
1 H NMR(400MHz，CD 3 OD)δ＝7.56(s，1H)，7.55-7.47(m，1H)，7.22-7.12(m，1H)，6.35(s，1H)，5.04(dd，J＝3.1，11.9Hz，1H)，4.53(dd，J＝3.0，12.0Hz，1H)，3.02(br d，J＝13.3Hz，1H)，2.81(s，3H)，2.50-2.40(m，2H)，2.39-2.27(m，2H)，2.22(br d，J＝7.0Hz，2H)，2.17-2.02(m，2H)，1.99(br d，J＝11.8Hz，2H)，1.24-1.10(m，2H)
Example 29
The synthetic route is as follows:
step 1: synthesis of Compound 42-a
To a thumb flask were added 20-b (50 mg, 109.27. Mu. Mol,1 eq), 2-fluoro-3-chloroaniline (23.86 mg, 163.91. Mu. Mol, 26.02. Mu. L,1.5 eq) and dichloromethane (1.5 mL) under nitrogen, and then pyridine (43.22 mg, 546.37. Mu. Mol, 44.10. Mu. L,5 eq) and phosphorus oxychloride (20.11 mg, 131.13. Mu. Mol, 12.19. Mu. L,1.2 eq) and the mixture was stirred at 0℃for 1 hour. The reaction solution was concentrated under reduced pressure to obtain compound 42-a.
MS m/z(ESI)：585.1[M+1] + .
Step 2: synthesis of Compound 42
In a thumb flask were added compound 42-a (60 mg, 102.54. Mu. Mol,1 eq) and methanol (1 mL), followed by sodium hydroxide (2M, 0.5mL,9.75 eq) and then stirred at 25℃for 1 hour. The reaction mixture was adjusted to ph=1 to 2 with 1M diluted hydrochloric acid and extracted with ethyl acetate (15 ml×3). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated by preparative high performance liquid chromatography (column form Phenomenex Gemini-NX C18 75 x 30mm x 3 μm; mobile phase [ water (0.225% formic acid) -acetonitrile ]; acetonitrile%: 35% -65%,7 min) to give compound 42.
MS m/z(ESI)：557.0[M+1] + .
1 H NMR(400MHz，CD 3 OD)δ＝7.75(ddd，J＝1.6，6.8，8.2Hz，1H)，7.57(s，1H)，7.35(ddd，J＝1.5，6.8，8.3Hz，1H)，7.19(dt，J＝1.6，8.2Hz，1H)，6.35(s，1H)，5.04(dd，J＝3.0，12.0Hz，1H)，4.53(dd，J＝3.0，12.0Hz，1H)，3.02(br d，J＝13.6Hz，1H)，2.82(s，3H)，2.53-2.29(m，4H)，2.25(d，J＝6.8Hz，2H)，2.18-1.96(m，4H)，1.27-1.07(m，2H).
Step 3: synthesis of Compounds 43 and 44
Compound 42 (25 mg, 44.88. Mu. Mol,1 eq) was purified by SFC (column: DAICEL CHIRALCEL OJ (250 mm. Times.30 mm,10 μm); mobile phase: [0.1% NH) 3 H 2 O，MeOH]Percent: 40% -40%) to give compounds 43 and 44.
SFC analysis method: column type: chiralcel OJ-3.150 x 4.6mm I.D.,3 μm; mobile phase: methanol (0.05% diethylamine); flow rate: 2.5mL/min.
Compound 43: MS m/z (ESI): 556.9[ M+1 ]] + SFC retention time: 3.996 min, ee% = 98.3%.
1 H NMR(400MHz，CD 3 OD)δ＝7.69-7.60(m，1H)，7.45(s，1H)，7.23(dt，J＝1.5，7.4Hz，1H)，7.07(dt，J＝1.6，8.2Hz，1H)，6.23(s，1H)，4.92(dd，J＝3.3，12.0Hz，1H)，4.41(dd，J＝3.0，12.0Hz，1H)，2.90(br d，J＝13.8Hz，1H)，2.82(s，3H)，2.38-2.16(m，4H)，2.12(d，J＝7.0Hz，2H)，2.05-1.83(m，4H)，1.14-0.92(m，2H).
Compound 44: MS m/z (ESI): 557.0[ M+1 ]] + SFC retention time: 5.221 min, ee% = 98.4%.
1 H NMR(400MHz，CD 3 OD)δ＝7.75(t，J＝7.4Hz，1H)，7.56(s，1H)，7.35(dt，J＝1.4，7.5Hz，1H)，7.19(dt，J＝1.5，8.2Hz，1H)，6.35(s，1H)，5.04(br d，J＝9.3Hz，1H)，4.53(dd，J＝2.8，12.0Hz，1H)，3.02(br d，J＝14.3Hz，1H)，2.82(s，3H)，2.51-2.39(m，2H)，2.39-2.28(m，2H)，2.24(d，J＝7.0Hz，2H)，2.18-2.08(m，1H)，2.05(dt，J＝3.5，7.3Hz，1H)，1.98(br d，J＝12.3Hz，2H)，1.26-1.06(m，2H).
Biological Activity test
Experimental example 1: quantitative qPCR assay for HBV in vitro test
1 purpose of experiment:
detection of HBV DNA content in HepG2.2.15 cells by real time quantitative qPCR assay (real time-qPCR) to compounds EC 50 The inhibition of HBV by the compounds was evaluated using the values as indicators.
2 experimental materials:
2.1 cell lines: hepG2.2.15 cells
HepG2.2.15 cell culture medium (DMEM/F12, invitrogen-11330057;10% serum, invitrogen-10099141;100units/ml penicillin and 10. Mu.g/ml streptomycin, invitrogen-15140122;1% nonessential amino acids, invitrogen-11140076;2mM L-glutamine, invitrogen-25030081; 300. Mu.g/ml geneticin, invitrogen-10131027).
2.2 reagents:
pancreatin (Invitrogen-25300062)
DPBS(Hyclone-SH30028.01B)
DMSO(Sigma-D2650-100ML)
High flux DNA purification kit (QIAamp 96 DNA Blood Kit,Qiagen-51162)
Quantitative quick-start universal probe reagent (FastStart Universal Probe Master, roche-04914058001)
2.3 consumables and instrumentation:
96-well cell culture plate (Coming-3599)
CO 2 Incubator (HERA-CELL-240)
Optical close plate film (ABI-4311971)
Quantitative PCR 96-well plate (Applied Biosystems-4306737)
Fluorescent quantitative PCR instrument (Applied Biosystems-7500 real time PCR system)
3. Experimental procedure and method:
3.1 HepG2.2.15 cells (4X 104 cells/well) to 96 well plates at 37℃with 5% CO 2 Culturing overnight.
3.2 the next day, the compound was diluted, total 8 concentrations, 3-fold gradient dilution. Adding different concentrations of compounds into the culture wells, and doubling the wells. The final concentration of DMSO in the culture was 1%.1 μM GLS4 served as 100% inhibition control; 1% DMSO was used as a 0% inhibition control.
3.3 day five, fresh medium containing the compound was changed.
3.4 culture broth from culture wells was harvested on day eight and DNA was extracted using a high throughput DNA purification kit (Qiagen-51162) for specific procedures with reference to the product instructions.
3.5 preparation of PCR reaction solution is shown in Table 1:
TABLE 1 preparation of PCR reaction solution
Upstream primer sequence: GTGTCTGCGGCGTTTTATCA
Downstream primer sequence: GACAAACGGGCAACATACCTT
Probe sequence: 5'+FAM+ CCTCTKCATCCTGCTGCTATGCCTCATC +TAMRA-3'
3.6 in 96 well PCR plate each well to 15. Mu.L of reaction mixture, then each well to 10. Mu.L of sample DNA or HBV DNA standard.
3.7 The reaction conditions for PCR were: heating at 95 ℃ for 10 minutes; then denatured at 95℃for 15 seconds and extended at 60℃for 1 minute for 40 cycles.
3.8 data analysis:
3.8.1 percent inhibition was calculated: % inh= [ 1- (copy number of DNA in sample-1 μm copy number of DNA in GLS 4)/(copy number of DNA in DMSO control-1 μm copy number of DNA in GLS 4) ]x100.
3.8.2 calculation of EC 50 : the 50% inhibitory concentration (EC) of compounds against HBV was calculated using GraphPad Prism software 50 ) Values.
4 experimental results are shown in table 2:
TABLE 2 qPCR assay to detect EC 50 Test results
Conclusion: the compound has remarkable inhibition effect on HBV.
Experimental example 2: hepatotoxicity test experiment
2.1: PHH cytotoxicity test experiment
1. Compounds were diluted in DMSO (dimethyl sulfoxide) at 3-fold gradient at 8 spots, double-multiplexed wells, and added to 96-well plates. The compound concentration was 200 times the final test concentration.
2. The cryopreserved human primary hepatocytes (PHH) were recovered, the cell density was counted with a cytometer, and the cell density was adjusted to the desired density with the medium.
3. Cells were added to 96-well plates to which the compounds had been added, with a final concentration of DMSO in each well of 0.5%. Cell wells containing 0.5% dmso were used as non-toxic negative controls, and cell broth wells were used as 100% cytotoxicity controls. The cell plates were then placed at 37℃in 5% CO 2 The cells were cultured in the incubator for 3 days.
4. The chemiluminescent signal (RLU, relative chemiluminescent unit) of each well in the cell plate was detected with a cell viability detection kit CellTiter-Glo, using a multifunctional enzyme-labeled instrument Synegy2-BioTek according to kit instructions.
5. The cell viability (cell viability%) of each well tested was calculated by substituting the raw data (RLU) into the following formula:
cell viability% = (RLU Sample -AverageRLU Mediumcontrol /(AverageRLU Cellcontrol -AverageRLU Mediumcontrol )×100％
RLU Sample Signal values for sample wells; averageRLU Cellcontrol Mean value of cell control well signal; averageRLU Mediumcontrol Mean value of medium control well signals.
6. Using GraphPad Prism software, non-linear fitting of cell viability data to a dose-response curve, and deriving the half-cytotoxicity concentration (CC 50 ) Values, results are shown in Table 3.
TABLE 3 half-cell toxicity concentration (CC 50 ) Value test results
| Compounds of formula (I) | CC 50 (μM) |
| Compound 11 | ＞100 |
Conclusion: cytotoxic CC of the inventive Compounds in Primary hepatocytes 50 The value is relatively high.
2.2: huh7 cytotoxicity test experiments
1. Compounds were diluted in DMSO (dimethyl sulfoxide) at 3-fold gradient at 8 spots, double-multiplexed wells, and added to 96-well plates. The compound concentration was 200 times the final test concentration.
2. Washing the cells with PBS (phosphate buffer), adding 0.25% pancreatin at 37deg.C, 5% CO 2 After digestion for about 2-5 minutes in the incubator, the digestion was stopped with cell culture medium and the cells were dispersed into single cells by blowing with a gun.
3. Cell densities were counted with a cytometer and adjusted to the desired densities with medium.
4. Cells were added to 96-well plates to which the compounds had been added, with a final concentration of DMSO in each well of 0.5%. Cell holes containing 0.5% DMSO are used as nontoxic negative control, and cell culture solution holes are used as 100%Cytotoxicity control. The cell plates were then placed at 37℃in 5% CO 2 The cells were cultured in the incubator for 3 days.
5. The chemiluminescent signal (RLU, relative chemiluminescent unit) of each well in the cell plate was detected with a cell viability detection kit CellTiter-Glo, using a multifunctional enzyme-labeled instrument Synegy2-BioTek according to kit instructions.
6. The cell viability (cell viability%) of each well tested was calculated by substituting the raw data (RLU) into the following formula:
Cell viability% = (RLU Sample -AverageRLU Mediumcontrol /(AverageRLU Cellcontrol -AverageRLU Mediumcontrol )×100％
RLU Sample Signal values for sample wells; averageRLU Cellcontrol Mean value of cell control well signal; averageRLU Mediumcontrol Mean value of medium control well signals.
7. Non-linear fitting of cell viability data to dose-response curves using GraphPad Prism software and derivation of half-cytotoxicity concentration of compounds (CC 50 ) Values, results are shown in table 4.
TABLE 4 half-cell toxicity concentration (CC 50 ) Value test results
| Compounds of formula (I) | CC 50 (μM) |
| Compound 11 | 86.71 |
Conclusion: cytotoxic CC of the compound in liver cancer cell Huh7 50 The value is relatively high.
2.3: hepG2 cytotoxicity test experiment
1. Compounds were diluted in DMSO (dimethyl sulfoxide) at 3-fold gradient at 8 spots, double-multiplexed wells, and added to 96-well plates. The compound concentration was 200 times the final test concentration.
2. Washing the cells with PBS (phosphate buffer), adding 0.25% pancreatin at 37deg.C, 5% CO 2 After digestion for about 2-5 minutes in the incubator, the digestion was stopped with cell culture medium and the cells were dispersed into single cells by blowing with a gun.
3. Cell densities were counted with a cytometer and adjusted to the desired densities with medium.
4. Cells were added to 96-well plates to which the compounds had been added, with a final concentration of DMSO in each well of 0.5%. Cell wells containing 0.5% dmso were used as non-toxic negative controls, and cell broth wells were used as 100% cytotoxicity controls. The cell plates were then placed at 37℃in 5% CO 2 The cells were cultured in the incubator for 3 days.
5. The chemiluminescent signal (RLU, relative chemiluminescent unit) of each well in the cell plate was detected with a cell viability detection kit CellTiter-Glo, using a multifunctional enzyme-labeled instrument Synegy2-BioTek according to kit instructions.
6. The cell viability (cell viability%) of each well tested was calculated by substituting the raw data (RLU) into the following formula:
cell viability% = (RLU Sample -AverageRLU Mediumcontrol /(AverageRLU Cellcontrol -AverageRLU Mediumcontrol )×100％
RLU Sample Signal values for sample wells; averageRLU Cellcontrol Mean value of cell control well signal; averageRLU Mediumcontrol Mean value of medium control well signals.
7. Nonlinear fitting of cell viability data to dose-response curves using GraphPad Prism softwareLine, and resulting in half-cytotoxicity concentration (CC 50 ) Values, results are shown in Table 5.
TABLE 5 half-cell toxicity concentration (CC 50 ) Value test results
| Compounds of formula (I) | CC 50 (μM) |
| Compound 11 | 79.35 |
Conclusion: cytotoxic CC of the compound in liver cancer cell HepG2 50 The value is relatively high.
Experimental example 3: pharmacokinetic studies
Pharmacokinetic study of Balb/c mice orally and intravenously injected test compounds:
the tested compound 11 is mixed with 10 percent polyethylene glycol-15 hydroxystearate/20 percent polyethylene glycol 400/70 percent aqueous solution, vortexed and sonicated to prepare 0.2mg/mL clear solution, and the clear solution is filtered by a microporous filter membrane for later use. Balb/c female mice 7 to 10 weeks old were selected and given a solution of the candidate compound by intravenous injection at a dose of 1mg/kg.
The tested compound 30 is mixed with 10% dimethyl sulfoxide/10% polyethylene glycol-15 hydroxystearate/80% aqueous solution, vortexed and sonicated to prepare a clear solution of 0.2mg/mL, and the clear solution is filtered through a microporous filter membrane for later use. Balb/c female mice 7 to 10 weeks old were selected and given a solution of the candidate compound by intravenous injection at a dose of 1mg/kg.
Test compound 36 was mixed with 10% dimethyl sulfoxide/10% polyethylene glycol-15 hydroxystearate/80% aqueous solution, vortexed and sonicated to prepare a clear solution of 0.2mg/mL, and filtered through a microfiltration membrane for use. Balb/c female mice 7 to 10 weeks old were selected and given a solution of the candidate compound by intravenous injection at a dose of 1mg/kg.
Test compounds 11, 30 and 36 were each mixed with 10% aqueous polyethylene glycol-15 hydroxystearate, vortexed and sonicated to prepare 1mg/mL homogeneous suspensions for use. Balb/c female mice 7 to 10 weeks old were selected and the candidate compound solution was orally administered at a dose of 10mg/kg.
Whole blood was collected for a certain period of time, plasma was prepared, drug concentration was analyzed by LC-MS/MS method, and drug substitution parameters were calculated by Phoenix WinNonlin software (Pharsight Co., USA) and the results are shown in Table 6.
TABLE 6 pharmacokinetic results of test compounds
Conclusion: the compound of the invention has high oral bioavailability.
Experimental example 4: in vivo liver-blood ratio study of mice
Liver-to-blood ratio study of oral test compounds in Balb/c mice
The compound is mixed with 10% polyethylene glycol-15 hydroxystearate aqueous solution, vortexed and sonicated to prepare 1mg/mL uniform suspension for use. Balb/c female mice 7 to 10 weeks old were selected and the candidate compound solution was orally administered at a dose of 10mg/kg.
Whole blood was collected for a certain period of time, plasma was prepared, liver tissue was collected for a corresponding period of time, tissue homogenate was prepared, drug concentration was analyzed by LC-MS/MS method, and drug substitution parameters were calculated by Phoenix WinNonlin software (Pharsight, usa). The results are shown in Table 7.
TABLE 7 liver to blood ratio results for test compounds
Conclusion: the ratio of the compounds of the present invention in liver and plasma is relatively high.
Experimental example 5: in vivo efficacy study
HDI/HBV model
The purpose of the experiment is as follows: the anti-hepatitis B virus effect of the compounds in mice was tested by the HDI/HBV mouse model.
Compound preparation: the solvent is 10% polyethylene glycol-15 hydroxystearate; a certain amount of test compounds 11, 30 and 36 were dissolved in 10% aqueous polyethylene glycol-15 hydroxystearate, respectively, vortexed and sonicated to prepare a uniform suspension, which was stored at 4 ℃ for use.
Mice were injected with HBV plasmid DNA solution at high pressure at tail vein: the day of plasmid injection was designated as day 0, day 1 after injection was designated as day 1, and so on. All animals were injected by tail vein with a physiological saline solution containing 10. Mu.g of plasmid DNA at 8% of the body weight on day 0, and the injection was completed within 5 seconds.
Administration: all animals were dosed twice daily by gavage (8/16 hours apart) on days 1-6, once on day 7, and all animals were euthanized at afternoon on day 7. The mice were monitored daily for body weight, which remained stable throughout the experiment.
Sample collection: all animals were collected on day 5 in the morning four hours following the first dose by submaxillary vein blood collection and plasma, all blood samples were collected on K 2 In EDTA anticoagulant tube, at 4℃7000g is centrifuged for 10 min to prepare about 40. Mu.L of plasma. Day 7 all animals were CO-dosed four hours after morning dosing 2 Euthanasia, heart blood sampling and plasma preparation method are the same. Two liver tissues, 70-100mg each, were collected and snap frozen with liquid nitrogen. After all samples were collected, they were stored in a-80℃refrigerator until dry ice conditions were reached and sent to the Ming Kangde department of biology in vitro laboratory for HBV DNA content detection.
Sample analysis: all plasma and liver samples were assayed for HBV DNA by qPCR.
Experimental results: the experimental results are shown in table 8.
TABLE 8 results of HDI efficacy experiments
Note that: deltaLog 10 copies represents the difference in viral load of the dosed group and the viral load of the vehicle group
Conclusion: the compound has remarkable effect of reducing HBV DNA in an HDI model.
Claims (19)
- A compound represented by the formula (I) or a pharmaceutically acceptable salt thereof,wherein,R 1 are each independently selected from halogen, OH, CN, NH 2 、C 1-3 Alkyl and C 1-3 Alkoxy group, the C 1-3 Alkyl and C 1-3 Alkoxy groups are each independently optionally substituted with 1, 2 or 3 halogens;m is selected from 0, 1, 2, 3 and 4;T 1 and T 2 Each independently selected from CH and N;is a single bond or a double bond;ring A is selected from phenyl, C 4-10 Cycloalkyl, the phenyl and C 4-10 Cycloalkyl groups are each independently optionally substituted with 1, 2Or 3R a Substitution;R a are each independently selected from halogen, OH, CN, NH 2 、C 1-3 Alkyl and C 1-3 Alkoxy group, the C 1-3 Alkyl and C 1-3 Alkoxy groups are each independently optionally substituted with 1, 2 or 3 halogens;L 1 selected from single bond, -C.ident.C-and-CR 2 ＝；R 2 Selected from H, methyl and F;L 2 selected from single bonds and methylene groups.
- The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R 1 Are respectively and independently selected from F, cl, br, CN and-CH 3 and-OCH 3 the-CH 3 and-OCH 3 Each independently optionally substituted with 1, 2 or 3F.
- The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein R 1 Each independently selected from F, cl, br and CN.
- The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein L 1 Selected from-c≡c-, -ch=, -C (F) =, and-C (CH) 3 )＝。
- The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R a Are respectively and independently selected from F, cl, br, CH 3 And OCH 3 The CH is 3 And OCH 3 Each independently optionally substituted with 1, 2 or 3F.
- The compound according to claim 1, or a pharmaceutically acceptable salt thereof, whereinStructural unitSelected from the group consisting of
- The compound of claim 6, or a pharmaceutically acceptable salt thereof, wherein the structural unitSelected from the group consisting of
- The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein ring a is selected from phenyl, cyclobutyl, cyclohexyl, The phenyl, cyclobutyl, cyclohexyl,Are each independently optionally substituted with 1, 2 or 3R a And (3) substitution.
- The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein ring a is selected from
- The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the structural unitSelected from the group consisting of
- A compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:Wherein R is 1 、R 2 、L 2 And m is as defined in any one of claims 1 to 5.
- The compound of claim 11, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:wherein R is 1 、R 2 、L 2 And m is as defined in claim 11.
- The compound of claim 12, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:wherein R is 1 、R 2 、L 2 And m is as defined in claim 12.
- A compound of the formula:
- the compound of claim 14, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:
- the compound of claim 15, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:
- the compound of claim 16, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:
- the use of a compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of a disease associated with hepatitis b virus infection.
- The use according to claim 18, wherein the disease associated with hepatitis b virus infection is chronic hepatitis b.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110494696.8 | 2021-05-07 | ||
| CN202110494696 | 2021-05-07 | ||
| PCT/CN2022/091012 WO2022233305A1 (en) | 2021-05-07 | 2022-05-05 | Methylsulfonylurea compound and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117337286A true CN117337286A (en) | 2024-01-02 |
Family
ID=83932008
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202280033410.9A Pending CN117337286A (en) | 2021-05-07 | 2022-05-05 | Methylsulfonyl urea compound and application thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN117337286A (en) |
| WO (1) | WO2022233305A1 (en) |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA3055194A1 (en) * | 2017-03-02 | 2018-09-07 | Assembly Biosciences, Inc. | Cyclic sulfamide compounds and methods of using same |
| WO2019206072A1 (en) * | 2018-04-24 | 2019-10-31 | 浙江海正药业股份有限公司 | Sulfamide aryl formamide derivative and preparation method therefor and uses thereof |
| EP3846820A4 (en) * | 2018-09-05 | 2022-01-19 | Assembly Biosciences, Inc. | Cyclic sulfamide compounds for treatment of hbv |
| US20220081433A1 (en) * | 2018-09-05 | 2022-03-17 | Assembly Biosciences, Inc. | Cyclic sulfamide compounds for treatment of hbv |
-
2022
- 2022-05-05 CN CN202280033410.9A patent/CN117337286A/en active Pending
- 2022-05-05 WO PCT/CN2022/091012 patent/WO2022233305A1/en active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| WO2022233305A1 (en) | 2022-11-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN115697338B (en) | Methyl substituted benzobisoxazole compound and application thereof | |
| JP7287978B2 (en) | two 4-{[(2S)-2-{4-[5-chloro-2-(1H-1,2,3-triazol-1-yl)phenyl]-5-methoxy-2-oxopyridine-1 Method for preparing (2H)-yl}butanoyl]amino}-2-fluorobenzamide derivatives | |
| JP7333342B2 (en) | 2,3-dihydro-1H-pyrrolidine-7-formamide derivative and use thereof | |
| US7910745B2 (en) | Organic compound and method for producing radioactive halogen-labeled organic compound using the same | |
| WO1992012991A1 (en) | Triterpene derivative | |
| RO111847B1 (en) | Optical izomer for the //4-(1,4,5,6-tetrahydro-4-methyl-6-oxo-3-pyridazinyl)-phenyl/hydr azono/propane dinitrile, preparation process thereof and intermediaries for the process making thereof | |
| CN116348466A (en) | Pyrazine sulfur biphenyl compound and application thereof | |
| CN114846008B (en) | Pyrimidine compound with fructokinase (KHK) inhibiting effect | |
| KR102345208B1 (en) | Quinolino-pyrrolidin-2-one derivatives and their applications | |
| TWI807697B (en) | Furan fused ring substituted glutarimide compounds | |
| WO2020052647A1 (en) | Spiro-heterocyclic compound acting as lsd1 inhibitor and use thereof | |
| WO1995022521A1 (en) | Aminoalkylcyclopropane derivative | |
| TW202317097A (en) | Glutarimide compounds and applications thereof | |
| CN117337278A (en) | Alkyl carboxylic acid compounds and uses thereof | |
| EP4105214A1 (en) | Use of jak inhibitors in preparation of drugs for treating jak kinase-related diseases | |
| CN117337286A (en) | Methylsulfonyl urea compound and application thereof | |
| TWI823274B (en) | Substituted pyridine-2,4-dione derivatives | |
| CN117279923A (en) | Derivative of six-membered heteroaromatic urea ring and application thereof | |
| CN115003672A (en) | Quinoline imidazole compound and application thereof | |
| PT92874A (en) | METHOD FOR PREPARING {(5-OXO-2-PYRROLIDINYL) -METHYL} -CYCLOHEXANOACETAMIDES AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM | |
| CN111777555B (en) | Aliphatic cycloalkane derivative, preparation method thereof, pharmaceutical composition and application thereof | |
| CN111057069B (en) | Cyclic compound, application and composition thereof | |
| TW202246279A (en) | Pyrazinethiobiphenyl compounds and application thereof | |
| WO2022143773A1 (en) | Benzimidazole compound and application thereof | |
| US8546424B2 (en) | Acetylene derivatives of 5-phenyl-pyrazolopyridine, preparation thereof, and therapeutic use thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |