WO2024027457A1 - Compound having ev71 and/or cva16 virus inhibitory activity and application thereof - Google Patents
Compound having ev71 and/or cva16 virus inhibitory activity and application thereof Download PDFInfo
- Publication number
- WO2024027457A1 WO2024027457A1 PCT/CN2023/106356 CN2023106356W WO2024027457A1 WO 2024027457 A1 WO2024027457 A1 WO 2024027457A1 CN 2023106356 W CN2023106356 W CN 2023106356W WO 2024027457 A1 WO2024027457 A1 WO 2024027457A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- compound
- pharmaceutically acceptable
- cva16
- prodrug
- solvate
- Prior art date
Links
- 150000001875 compounds Chemical class 0.000 title claims abstract description 66
- 230000002401 inhibitory effect Effects 0.000 title abstract description 19
- 241000700605 Viruses Species 0.000 title description 36
- 241001529459 Enterovirus A71 Species 0.000 claims abstract description 88
- 239000003814 drug Substances 0.000 claims abstract description 23
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 19
- 201000010099 disease Diseases 0.000 claims abstract description 18
- 229940079593 drug Drugs 0.000 claims abstract description 18
- 241001429382 Coxsackievirus A16 Species 0.000 claims abstract 7
- 150000003839 salts Chemical class 0.000 claims description 28
- 229940125904 compound 1 Drugs 0.000 claims description 19
- 239000012453 solvate Substances 0.000 claims description 18
- 239000000651 prodrug Substances 0.000 claims description 17
- 229940002612 prodrug Drugs 0.000 claims description 17
- 239000008194 pharmaceutical composition Substances 0.000 claims description 13
- 239000000243 solution Substances 0.000 claims description 13
- 229940126214 compound 3 Drugs 0.000 claims description 11
- -1 inhalants Substances 0.000 claims description 11
- 125000004169 (C1-C6) alkyl group Chemical group 0.000 claims description 10
- 150000002431 hydrogen Chemical class 0.000 claims description 10
- 229910052739 hydrogen Inorganic materials 0.000 claims description 10
- 239000001257 hydrogen Substances 0.000 claims description 10
- 239000000825 pharmaceutical preparation Substances 0.000 claims description 10
- 208000007212 Foot-and-Mouth Disease Diseases 0.000 claims description 8
- 241000710198 Foot-and-mouth disease virus Species 0.000 claims description 8
- 229910052736 halogen Inorganic materials 0.000 claims description 8
- 150000002367 halogens Chemical class 0.000 claims description 8
- 229940125782 compound 2 Drugs 0.000 claims description 7
- 230000003612 virological effect Effects 0.000 claims description 7
- 125000000171 (C1-C6) haloalkyl group Chemical group 0.000 claims description 6
- YZCKVEUIGOORGS-OUBTZVSYSA-N Deuterium Chemical compound [2H] YZCKVEUIGOORGS-OUBTZVSYSA-N 0.000 claims description 6
- 229910052805 deuterium Inorganic materials 0.000 claims description 6
- 125000004178 (C1-C4) alkyl group Chemical group 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 239000000725 suspension Substances 0.000 claims description 4
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 claims description 2
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 claims description 2
- 229910052794 bromium Inorganic materials 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 125000000753 cycloalkyl group Chemical group 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 239000000499 gel Substances 0.000 claims description 2
- 239000008187 granular material Substances 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 claims description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 239000006187 pill Substances 0.000 claims description 2
- 239000000829 suppository Substances 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims description 2
- 230000000155 isotopic effect Effects 0.000 claims 3
- 230000010076 replication Effects 0.000 abstract description 20
- 102000004169 proteins and genes Human genes 0.000 abstract description 12
- 108090000623 proteins and genes Proteins 0.000 abstract description 12
- 230000002265 prevention Effects 0.000 abstract description 7
- 238000011282 treatment Methods 0.000 abstract description 5
- 239000003443 antiviral agent Substances 0.000 abstract description 2
- 230000029812 viral genome replication Effects 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 57
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 24
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- 238000006243 chemical reaction Methods 0.000 description 13
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 12
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 11
- 238000000034 method Methods 0.000 description 11
- 239000007787 solid Substances 0.000 description 11
- 101710132601 Capsid protein Proteins 0.000 description 10
- 101710197658 Capsid protein VP1 Proteins 0.000 description 10
- 101710118046 RNA-directed RNA polymerase Proteins 0.000 description 10
- 101710108545 Viral protein 1 Proteins 0.000 description 10
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 9
- 108020000999 Viral RNA Proteins 0.000 description 9
- 208000015181 infectious disease Diseases 0.000 description 9
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 8
- 230000005764 inhibitory process Effects 0.000 description 8
- 239000006228 supernatant Substances 0.000 description 8
- 238000005481 NMR spectroscopy Methods 0.000 description 7
- 230000030833 cell death Effects 0.000 description 7
- 239000002904 solvent Substances 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 238000001262 western blot Methods 0.000 description 7
- 108020004684 Internal Ribosome Entry Sites Proteins 0.000 description 6
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 6
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 6
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 6
- 230000002829 reductive effect Effects 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 5
- 238000004440 column chromatography Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 239000012074 organic phase Substances 0.000 description 5
- 230000007170 pathology Effects 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 208000024891 symptom Diseases 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 108091005804 Peptidases Proteins 0.000 description 4
- 239000004365 Protease Substances 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 239000003208 petroleum Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 3
- WMFOQBRAJBCJND-UHFFFAOYSA-M Lithium hydroxide Chemical compound [Li+].[OH-] WMFOQBRAJBCJND-UHFFFAOYSA-M 0.000 description 3
- SECXISVLQFMRJM-UHFFFAOYSA-N N-Methylpyrrolidone Chemical compound CN1CCCC1=O SECXISVLQFMRJM-UHFFFAOYSA-N 0.000 description 3
- 108700026244 Open Reading Frames Proteins 0.000 description 3
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- 108010076039 Polyproteins Proteins 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 210000003169 central nervous system Anatomy 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- UAOMVDZJSHZZME-UHFFFAOYSA-N diisopropylamine Chemical compound CC(C)NC(C)C UAOMVDZJSHZZME-UHFFFAOYSA-N 0.000 description 3
- 238000010166 immunofluorescence Methods 0.000 description 3
- 230000002441 reversible effect Effects 0.000 description 3
- 239000003981 vehicle Substances 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 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 2
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 2
- 241000709661 Enterovirus Species 0.000 description 2
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 2
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 2
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 241000709664 Picornaviridae Species 0.000 description 2
- 108010001267 Protein Subunits Proteins 0.000 description 2
- 102000002067 Protein Subunits Human genes 0.000 description 2
- 101800001554 RNA-directed RNA polymerase Proteins 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 101710172711 Structural protein Proteins 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 108091023045 Untranslated Region Proteins 0.000 description 2
- 108010067390 Viral Proteins Proteins 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- 210000000133 brain stem Anatomy 0.000 description 2
- 210000000234 capsid Anatomy 0.000 description 2
- 239000008004 cell lysis buffer Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 231100000673 dose–response relationship Toxicity 0.000 description 2
- 241001493065 dsRNA viruses Species 0.000 description 2
- 230000001747 exhibiting effect Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- JJWLVOIRVHMVIS-UHFFFAOYSA-N isopropylamine Chemical compound CC(C)N JJWLVOIRVHMVIS-UHFFFAOYSA-N 0.000 description 2
- IYRMWMYZSQPJKC-UHFFFAOYSA-N kaempferol Chemical compound C1=CC(O)=CC=C1C1=C(O)C(=O)C2=C(O)C=C(O)C=C2O1 IYRMWMYZSQPJKC-UHFFFAOYSA-N 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000012299 nitrogen atmosphere Substances 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 150000007530 organic bases Chemical class 0.000 description 2
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 2
- VDZOOKBUILJEDG-UHFFFAOYSA-M tetrabutylammonium hydroxide Chemical compound [OH-].CCCC[N+](CCCC)(CCCC)CCCC VDZOOKBUILJEDG-UHFFFAOYSA-M 0.000 description 2
- CZDYPVPMEAXLPK-UHFFFAOYSA-N tetramethylsilane Chemical compound C[Si](C)(C)C CZDYPVPMEAXLPK-UHFFFAOYSA-N 0.000 description 2
- 238000004809 thin layer chromatography Methods 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
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 2
- 230000014616 translation Effects 0.000 description 2
- 238000013519 translation Methods 0.000 description 2
- 230000006656 viral protein synthesis Effects 0.000 description 2
- NKRAIOQPSBRMOV-NRMKKVEVSA-N (2r,3r,4r,5r)-2-(4-aminopyrrolo[2,3-d]pyrimidin-7-yl)-3-ethynyl-5-(hydroxymethyl)oxolane-3,4-diol Chemical compound C1=CC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@]1(O)C#C NKRAIOQPSBRMOV-NRMKKVEVSA-N 0.000 description 1
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- FNMASAFFZKWPGI-UHFFFAOYSA-N 1-[5-(4-bromophenoxy)pentyl]-3-pyridin-4-ylimidazolidin-2-one Chemical compound C1=CC(Br)=CC=C1OCCCCCN1C(=O)N(C=2C=CN=CC=2)CC1 FNMASAFFZKWPGI-UHFFFAOYSA-N 0.000 description 1
- QNTITXYHUMSSKB-UHFFFAOYSA-N 1-pyridin-2-ylimidazolidin-2-one Chemical class O=C1NCCN1C1=CC=CC=N1 QNTITXYHUMSSKB-UHFFFAOYSA-N 0.000 description 1
- BTLKROSJMNFSQZ-UHFFFAOYSA-N 2,4-dichloro-6-methylpyrimidine Chemical compound CC1=CC(Cl)=NC(Cl)=N1 BTLKROSJMNFSQZ-UHFFFAOYSA-N 0.000 description 1
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 description 1
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- BWGRDBSNKQABCB-UHFFFAOYSA-N 4,4-difluoro-N-[3-[3-(3-methyl-5-propan-2-yl-1,2,4-triazol-4-yl)-8-azabicyclo[3.2.1]octan-8-yl]-1-thiophen-2-ylpropyl]cyclohexane-1-carboxamide Chemical compound CC(C)C1=NN=C(C)N1C1CC2CCC(C1)N2CCC(NC(=O)C1CCC(F)(F)CC1)C1=CC=CS1 BWGRDBSNKQABCB-UHFFFAOYSA-N 0.000 description 1
- ALMBOXQFPLQVLF-UHFFFAOYSA-N 4-chloro-6-methyl-2-methylsulfanylpyrimidine Chemical compound CSC1=NC(C)=CC(Cl)=N1 ALMBOXQFPLQVLF-UHFFFAOYSA-N 0.000 description 1
- 108020003589 5' Untranslated Regions Proteins 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 238000003734 CellTiter-Glo Luminescent Cell Viability Assay Methods 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 241000709687 Coxsackievirus Species 0.000 description 1
- UBSCDKPKWHYZNX-UHFFFAOYSA-N Demethoxycapillarisin Natural products C1=CC(O)=CC=C1OC1=CC(=O)C2=C(O)C=C(O)C=C2O1 UBSCDKPKWHYZNX-UHFFFAOYSA-N 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 1
- 241000991587 Enterovirus C Species 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 108091026898 Leader sequence (mRNA) Proteins 0.000 description 1
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 208000009525 Myocarditis Diseases 0.000 description 1
- LFZAGIJXANFPFN-UHFFFAOYSA-N N-[3-[4-(3-methyl-5-propan-2-yl-1,2,4-triazol-4-yl)piperidin-1-yl]-1-thiophen-2-ylpropyl]acetamide Chemical compound C(C)(C)C1=NN=C(N1C1CCN(CC1)CCC(C=1SC=CC=1)NC(C)=O)C LFZAGIJXANFPFN-UHFFFAOYSA-N 0.000 description 1
- 239000007832 Na2SO4 Substances 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 239000012425 OXONE® Substances 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 1
- 206010063837 Reperfusion injury Diseases 0.000 description 1
- IWUCXVSUMQZMFG-AFCXAGJDSA-N Ribavirin Chemical compound N1=C(C(=O)N)N=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 IWUCXVSUMQZMFG-AFCXAGJDSA-N 0.000 description 1
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- 108091036066 Three prime untranslated region Proteins 0.000 description 1
- ABRVLXLNVJHDRQ-UHFFFAOYSA-N [2-pyridin-3-yl-6-(trifluoromethyl)pyridin-4-yl]methanamine Chemical compound FC(C1=CC(=CC(=N1)C=1C=NC=CC=1)CN)(F)F ABRVLXLNVJHDRQ-UHFFFAOYSA-N 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 229910052783 alkali metal Inorganic materials 0.000 description 1
- 229910052784 alkaline earth metal Inorganic materials 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 239000004599 antimicrobial Substances 0.000 description 1
- 230000008335 axon cargo transport Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- SRSXLGNVWSONIS-UHFFFAOYSA-N benzenesulfonic acid Chemical compound OS(=O)(=O)C1=CC=CC=C1 SRSXLGNVWSONIS-UHFFFAOYSA-N 0.000 description 1
- 229940092714 benzenesulfonic acid Drugs 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000008499 blood brain barrier function Effects 0.000 description 1
- 210000001218 blood-brain barrier Anatomy 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 235000010216 calcium carbonate Nutrition 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 210000003591 cerebellar nuclei Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 229940121657 clinical drug Drugs 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 229940043279 diisopropylamine Drugs 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 206010014599 encephalitis Diseases 0.000 description 1
- CAYJBRBGZBCZKO-BHGBQCOSSA-N ethyl (e,4s)-4-[[(2r,5s)-2-[(4-fluorophenyl)methyl]-6-methyl-5-[(5-methyl-1,2-oxazole-3-carbonyl)amino]-4-oxoheptanoyl]amino]-5-[(3s)-2-oxopyrrolidin-3-yl]pent-2-enoate Chemical compound C([C@@H](/C=C/C(=O)OCC)NC(=O)[C@@H](CC(=O)[C@@H](NC(=O)C1=NOC(C)=C1)C(C)C)CC=1C=CC(F)=CC=1)[C@@H]1CCNC1=O CAYJBRBGZBCZKO-BHGBQCOSSA-N 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 210000004884 grey matter Anatomy 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- IXCSERBJSXMMFS-UHFFFAOYSA-N hydrogen chloride Substances Cl.Cl IXCSERBJSXMMFS-UHFFFAOYSA-N 0.000 description 1
- 229910000041 hydrogen chloride Inorganic materials 0.000 description 1
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 description 1
- 229940071870 hydroiodic acid Drugs 0.000 description 1
- 210000003016 hypothalamus Anatomy 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 150000007529 inorganic bases Chemical class 0.000 description 1
- 208000028867 ischemia Diseases 0.000 description 1
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 description 1
- 235000008777 kaempferol Nutrition 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 229910052744 lithium Inorganic materials 0.000 description 1
- ZLNQQNXFFQJAID-UHFFFAOYSA-L magnesium carbonate Chemical compound [Mg+2].[O-]C([O-])=O ZLNQQNXFFQJAID-UHFFFAOYSA-L 0.000 description 1
- 239000001095 magnesium carbonate Substances 0.000 description 1
- 229910000021 magnesium carbonate Inorganic materials 0.000 description 1
- 235000014380 magnesium carbonate Nutrition 0.000 description 1
- 159000000003 magnesium salts Chemical class 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- 239000011976 maleic acid Substances 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 229940098779 methanesulfonic acid Drugs 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- UXOUKMQIEVGVLY-UHFFFAOYSA-N morin Natural products OC1=CC(O)=CC(C2=C(C(=O)C3=C(O)C=C(O)C=C3O2)O)=C1 UXOUKMQIEVGVLY-UHFFFAOYSA-N 0.000 description 1
- 230000007171 neuropathology Effects 0.000 description 1
- 230000002276 neurotropic effect Effects 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 210000000633 nuclear envelope Anatomy 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- PIBWKRNGBLPSSY-UHFFFAOYSA-L palladium(II) chloride Chemical compound Cl[Pd]Cl PIBWKRNGBLPSSY-UHFFFAOYSA-L 0.000 description 1
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- HJKYXKSLRZKNSI-UHFFFAOYSA-I pentapotassium;hydrogen sulfate;oxido sulfate;sulfuric acid Chemical compound [K+].[K+].[K+].[K+].[K+].OS([O-])(=O)=O.[O-]S([O-])(=O)=O.OS(=O)(=O)O[O-].OS(=O)(=O)O[O-] HJKYXKSLRZKNSI-UHFFFAOYSA-I 0.000 description 1
- 210000000578 peripheral nerve Anatomy 0.000 description 1
- 239000011736 potassium bicarbonate Substances 0.000 description 1
- 229910000028 potassium bicarbonate Inorganic materials 0.000 description 1
- 235000015497 potassium bicarbonate Nutrition 0.000 description 1
- 229910000027 potassium carbonate Inorganic materials 0.000 description 1
- 235000011181 potassium carbonates Nutrition 0.000 description 1
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 description 1
- 159000000001 potassium salts Chemical class 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 229960000329 ribavirin Drugs 0.000 description 1
- HZCAHMRRMINHDJ-DBRKOABJSA-N ribavirin Natural products O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1N=CN=C1 HZCAHMRRMINHDJ-DBRKOABJSA-N 0.000 description 1
- 229950007656 rupintrivir Drugs 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- WRIKHQLVHPKCJU-UHFFFAOYSA-N sodium bis(trimethylsilyl)amide Chemical compound C[Si](C)(C)N([Na])[Si](C)(C)C WRIKHQLVHPKCJU-UHFFFAOYSA-N 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 235000017550 sodium carbonate Nutrition 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 210000000278 spinal cord Anatomy 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- 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 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 210000002845 virion Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
- C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
- C07D495/04—Ortho-condensed systems
Definitions
- the invention belongs to the field of antiviral drugs and relates to a compound with EV71 and/or CVA16 virus inhibitory activity and its application in the preparation of anti-EV71 and/or CVA16 virus drugs and its use in medicine, especially for prevention and/or for the treatment of conditions associated with EV71 and/or CVA16 viruses, including hand, foot and mouth disease.
- EV71 Human enterovirus 71
- CVA16 coxsackievirus group A 16
- EV71 can cause severe infection of the central nervous system, and in severe cases, death of individuals.
- CVA16 causes milder clinical symptoms, but can also cause serious complications, such as pneumonia, encephalitis, and myocarditis (Pediatr Infect Dis J. 2004 Mar; 23(3):276-8.).
- EV71 is a single-stranded positive-strand RNA virus, belonging to the Picornaviridae family and enterovirus class A (Nat Struct Mol Biol. 19 (2012) 424-429).
- Both EV71 and CVA16 are single-stranded positive-sense RNA viruses without a nuclear membrane. They are an icosahedron with a diameter of 20-30nm. They belong to the Picornaviridae family and are subordinate to enteroviruses. Its full genome size is approximately 7400bp, consisting of a 5' untranslated region (UTR), a 3' untranslated region (UTR), and an open reading frame (ORF), encoding a polyprotein.
- the open reading frame can be divided into three sub-structures, called P1, P2 and P3 regions.
- the P1 region mainly encodes four structural proteins, divided into VP1, VP2, VP3 and VP4.
- VP1, VP2 and VP3 are on the outside of the virus capsid and can serve as viral antigenic determinants, and VP4 is located on the inside of the virus capsid (Nature. 317 (1985) 145-153).
- the P2 region encodes three nonstructural proteins, divided into 2A, 2B and 2C.
- the P3 region encodes four nonstructural proteins, divided into 3A, 3B, 3C and 3D.
- EV71 virus can cleave the mature polyprotein of the virus.
- 3D protein is RNA polymerase and plays a vital role in the process of viral translation (The Lancet Infectious Diseases. 10 (2010) 778-790; Journal of Biomedical Science. 21 (2014) 1-14).
- the EV71 5'UTR This region contains an internal ribosome entry site (IRES), which replaces the eukaryotic 5' cap structure and initiates the viral translation process.
- IRES internal ribosome entry site
- EV71 is a highly neurotropic virus, and the most common target of EV71 infection is the brainstem (The New England Journal of Medicine. 341 (1999) 936-942).
- the infection route of EV71 and CVA16 is very similar to that of poliovirus.
- EV71 can cause infection in the spinal cord gray matter, brainstem, hypothalamus and dentate nucleus, and inflammation is the most obvious infection symptom (Journal of Neuropathology and Experimental Neurology. 67 (2008) 162-169).
- EV71 proteases 2A and 3C can cleave the maturation of EV71 polyprotein and are crucial for the function of EV71.
- Rupintrivir can inhibit EV71 replication by inhibiting the activity of EV71 3C protease (Journal of Virology. 85 (2011) 10319-10331). 3. Key proteins that inhibit EV71 replication.
- EV71 RNA genome replication depends on EV71 RNA polymerase 3D, so targeting the 3D protein can effectively inhibit EV71 replication. Nucleotide analogs, ribavirin and NITD008, and non-nucleotide analogs, DTriP-22, have been shown to inhibit 3D polymerase (J Am Coll Cardiol.
- EV71 infection has become a very serious public safety issue, especially in the Asia-Pacific region. Since the clinical symptoms caused by EV71 are more severe than those caused by CVA16, most scientific research teams prefer to develop anti-EV71 drugs, while there are fewer studies on anti-CVA16 drugs. Currently, there are no clinical treatments for disease caused by EV71 and/or CVA16 infection Therefore, it is imperative to develop drugs that are effective against EV71 and/or CVA16 viruses.
- the object of the present invention is to provide a compound with EV71 and/or CVA16 virus inhibitory activity and its application in the preparation of anti-EV71 and/or CVA16 virus drugs.
- the present invention provides a compound with EV71 and/or CVA16 virus inhibitory activity or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof.
- the compound has the structure of formula (I):
- n is any integer from 1 to 6; preferably 1;
- n 0, 1 or 2; preferably 1;
- R 1 is each independently hydrogen, deuterium, halogen, C1-C6 alkyl, C1-C6 haloalkyl or
- R 2 is each independently hydrogen, deuterium, halogen, C1-C6 alkyl or C1-C6 haloalkyl;
- R 3 is C1-C6 alkyl, C1-C6 haloalkyl or C4-C6 cycloalkyl.
- the compound has the structure of formula (II):
- R 1 , R 2 and R 3 are as defined in formula (I).
- R1 is hydrogen, deuterium, halogen or More preferably hydrogen, halogen or Further preference is given to hydrogen, bromine or
- R 2 is C1-C6 alkyl, more preferably C1-C4 alkyl, further preferably methyl.
- R 3 is C1-C6 alkyl, more preferably C1-C4 alkyl, further preferably isopropyl.
- compound 1 is compound 1, compound 2 or compound 3:
- the present invention provides a pharmaceutical composition
- a pharmaceutical composition comprising the compound according to the ⁇ first aspect> or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof.
- the pharmaceutical composition further contains pharmaceutically acceptable excipients or carriers.
- the present invention provides a pharmaceutical preparation, which contains a compound according to the ⁇ first aspect> or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof, or a compound according to the ⁇ second aspect> >The pharmaceutical composition.
- the pharmaceutical preparation is any one of tablets, capsules, injections, granules, powders, suppositories, pills, gels, powders, oral solutions, inhalants, suspensions or dry suspensions.
- the present invention provides a pharmaceutical combination form, which contains a compound according to the ⁇ first aspect> or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof, or a compound according to the ⁇ second aspect> Aspect>
- the present invention provides a compound according to the ⁇ first aspect> or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof, or a pharmaceutical composition according to the ⁇ second aspect> , or according to the pharmaceutical preparation described in the ⁇ third aspect>, or according to the application of the pharmaceutical combination form described in the ⁇ fourth aspect> in the preparation of anti-EV71 and/or CVA16 viral drugs.
- the medicament is a medicament for the prevention and/or treatment of diseases and/or conditions that are at least partially responsive to EV71 and/or CVA16 viruses.
- the drug is a drug for preventing and/or treating hand, foot and mouth disease.
- the present invention provides a compound according to the ⁇ first aspect> or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof, or a pharmaceutical composition according to the ⁇ second aspect> , or the pharmaceutical preparation according to the ⁇ third aspect>, or the pharmaceutical combination form according to the ⁇ fourth aspect>, which is used to prevent and/or treat diseases and/or at least partially responsive to EV71 and/or CVA16 viruses or illness.
- the disease and/or condition that is at least partially responsive to EV71 and/or CVA16 viruses is hand, foot and mouth disease.
- the present invention provides a method for preventing and/or treating diseases and/or conditions that are at least partially responsive to EV71 and/or CVA16 viruses, which includes the following steps: adding a preventive and/or therapeutically effective amount of a drug according to ⁇ first aspect >The compound or its pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer, or the pharmaceutical composition according to the ⁇ second aspect>, or the pharmaceutical composition according to the ⁇ third aspect>.
- the pharmaceutical preparation described above, or the pharmaceutical combination described in the ⁇ Fourth Aspect> is administered to individuals in need thereof.
- the disease and/or condition that is at least partially responsive to EV71 and/or CVA16 viruses is hand, foot and mouth disease.
- the compound provided by the invention has good inhibitory activity against EV71 and/or CVA16 viruses, can inhibit the replication and protein expression of EV71 and/or CVA16 viruses in host cells, and has a low half inhibitory concentration, and has the potential to be used as an anti-EV71 and/or CVA16 virus.
- CVA16 virus drugs used for clinical prevention and/or treatment caused by EV71 and/or CVA16 viruses diseases and/or conditions.
- Figure 1 is a scatter plot showing the inhibition of EV71 virus titer by compounds 1 and 2 in Example 4.
- Figure 2 is a bar graph of the half-inhibition rate of compounds 1 and 2 in Example 4 in inhibiting cell death caused by EV71 virus.
- Figure 3 is a bar graph showing that compound 3 inhibits cell death caused by EV71 virus in Example 4.
- Figure 4 is a bar graph showing the inhibition of EV71 viral RNA replication by compounds 1 and 2 in Example 5.
- Figure 5 is a bar graph showing the inhibition of EV71 viral RNA replication by Compound 3 in Example 5.
- Figure 6 is an agarose gel image showing the inhibition of EV71 viral RNA replication by compounds 1 and 2 in Example 5.
- Figure 7 is an immunofluorescence graph showing the inhibition of EV71 virus-specific 3D expression by Compounds 1 and 2 in Example 6.
- Figure 8 is a Western blot diagram showing the inhibition of EV71 virus 3D and VP1 expression by Compounds 1 and 2 at different concentrations in Example 6.
- Figure 9 is a Western blot diagram showing the inhibition of EV71 virus 3D and VP1 expression by Compound 3 at different concentrations in Example 6.
- Figure 10 is a Western blot diagram showing the inhibition of EV71 virus 3D and VP1 expression by Compounds 1 and 2 at different time periods in Example 6.
- Figure 11 is a bar graph showing that Compound 1 inhibits cell death caused by CVA16 in Example 7.
- Figure 12 is a bar graph of compound 1 inhibiting CVA16 viral RNA replication in Example 8.
- Figure 13 is a Western blot diagram showing that Compound 1 inhibits the expression of CVA16 virus 3D protein in Example 9.
- pharmaceutically acceptable salts means that the compounds of the present invention exist in the form of their pharmaceutically acceptable salts, including acid addition salts and base addition salts.
- Pharmaceutically acceptable salts are described in Pharmaceutical Salts by SM Berge in J. Pharmaceutical Sciences (Vol. 66: pp. 1-19, 1977).
- pharmaceutically acceptable non-toxic acid addition salts refer to salts formed by compounds in the present invention and organic or inorganic acids, including (but not limited to) hydrochloric acid, sulfuric acid, hydrobromic acid, Hydroiodic acid, phosphoric acid, nitric acid, perchloric acid, acetic acid, oxalic acid, Maleic acid, fumaric acid, tartaric acid, benzenesulfonic acid, methanesulfonic acid, salicylic acid, succinic acid, citric acid, lactic acid, propionic acid, benzoic acid, p-toluenesulfonic acid and malic acid, etc.
- organic or inorganic acids including (but not limited to) hydrochloric acid, sulfuric acid, hydrobromic acid, Hydroiodic acid, phosphoric acid, nitric acid, perchloric acid, acetic acid, oxalic acid, Maleic acid, fumaric acid, tartaric acid, benzenesulfonic
- Non-toxic base addition salts represent salts formed by compounds in the present invention and organic or inorganic bases, including (but not limited to) alkali metal salts, such as lithium, sodium or potassium salts; alkaline earth metal salts, For example, calcium or magnesium salts; organic base salts, such as ammonium salts or N + (C 1-6 alkyl) 4 salts formed with organic bases containing N groups, preferably lithium hydroxide, sodium hydroxide, hydroxide Potassium, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, magnesium carbonate, calcium carbonate, ammonia, triethylamine or tetrabutylammonium hydroxide, etc.
- “Pharmaceutically acceptable salts” can be synthesized by general chemical methods.
- solvate refers to an association of one or more solvent molecules with a compound of the present invention.
- Solvents that form solvates include, but are not limited to, water, methanol, ethanol, isopropyl alcohol, ethyl acetate, tetrahydrofuran, N,N-dimethylformamide, dimethyl sulfoxide, and the like.
- prodrug refers to a chemical derivative of a compound of the present invention that is converted into a compound of the present invention by a chemical reaction in the body.
- isotope label means isotopes including (but not limited to) 2 H, 3 H, 11 C, 13 C, 14 C, 15 N, etc.
- isomers includes all isomeric forms, including enantiomers, diastereomers, tautomers and geometric isomers (including cis and trans isomers). Therefore, the individual stereochemical isomers or enantiomers, diastereomers, tautomers or geometric isomers (or cis-trans isomers) of the compounds designed in the present invention Mixtures are within the scope of the invention.
- prevention means, for example, the complete or almost complete prevention of the occurrence of a disease or condition (e.g., infection, ischemia or reperfusion injury) when a patient or subject is susceptible to the condition or at risk of the disease or condition; Prevention can also include suppression, that is, halting the progression of the condition.
- a disease or condition e.g., infection, ischemia or reperfusion injury
- treating means: 1) inhibiting the disease; e.g., inhibiting the disease, condition, or condition in an individual who is experiencing or exhibiting pathology or symptomatology of the disease, condition, or condition (i.e., preventing the pathology and/or symptoms or 2) ameliorating the disease; e.g., ameliorating the disease, condition, or condition in an individual who is experiencing or exhibiting the pathology or symptomatology of the disease, condition, or disorder (i.e., reversing the pathology and/or symptoms study).
- DMSO dimethyl sulfoxide
- Compound 1 was synthesized by the following method:
- Compound 2 was synthesized by the following method:
- Example 5 Test of the inhibitory ability of compounds 1, 2 and 3 on EV71 virus replication
- Example 6 Test of the inhibitory ability of compounds 1, 2 and 3 on EV71 viral protein expression
- RD cells were pretreated with different concentrations of compound 1, 2 or 3 for 1 hour;
- Example 7 Test of the ability of compound 1 to inhibit cell death caused by CVA16 virus
- Example 8 Test of the ability of compound 1 to inhibit CVA16 viral RNA replication
- Compound 1 can effectively reduce the replication of CVA16 viral RNA in a dose-dependent manner within a certain range.
- Example 9 Test of the ability of compound 1 to inhibit the expression of CVA16 viral protein
- the compounds of the present invention can effectively inhibit the replication of EV71 and/or CVA16 in host cells, and have the potential to be used as clinical drugs for preventing and/or treating diseases and/or conditions caused by EV71 and/or CVA16.
Abstract
A compound which has EV71 and/or CVA16 virus inhibitory activity and an application thereof, belonging to the field of antiviral drugs. The compound has the structure of formula (I), has good EV71 and/or CVA16 virus replication inhibitory activity, can effectively inhibit the replication and protein expression of EV71 and/or CVA16 viruses in host cells, and has the potential to be used as an anti-EV71 and/or CVA16 virus drug for the prevention and/or treatment of diseases and/or conditions caused by EV71 and/or CVA16 viruses.
Description
相关申请的引用References to related applications
本发明要求2022年08月04日在中国提交的，名称为“具有EV71和/或CVA16病毒抑制活性的化合物及其应用”、申请号为202210933895.9的发明专利申请的优先权，通过引用的方式将上述专利申请的全部内容并入本文。The present invention claims the priority of the invention patent application titled "Compounds with EV71 and/or CVA16 virus inhibitory activity and applications thereof" and application number 202210933895.9, submitted in China on August 4, 2022, and shall be incorporated by reference. The entire contents of the above patent application are incorporated herein.
本发明属于抗病毒药物领域，涉及一种具有EV71和/或CVA16病毒抑制活性的化合物及其在制备抗EV71和/或CVA16病毒药物中的应用及其在医学中的用途，特别是用于预防和/或治疗与EV71和/或CVA16病毒相关的病症(包括手足口病)的用途。The invention belongs to the field of antiviral drugs and relates to a compound with EV71 and/or CVA16 virus inhibitory activity and its application in the preparation of anti-EV71 and/or CVA16 virus drugs and its use in medicine, especially for prevention and/or for the treatment of conditions associated with EV71 and/or CVA16 viruses, including hand, foot and mouth disease.
人肠道病毒71型(EV71)和柯萨奇病毒A组16型(CVA16)是人类手足口病的主要病原体。其中，EV71可以引起严重的中枢神经系统的感染，严重者可引起个体死亡。与EV71相比，CVA16引起的临床症状较轻，但也可引起严重的并发症，如肺炎、脑炎和心肌炎等(Pediatr Infect Dis J.2004 Mar；23(3):276-8.)。EV71为单股正链RNA病毒，属于小核糖核酸病毒科，从属于肠道病毒A类(Nat Struct Mol Biol.19(2012)424-429)。Human enterovirus 71 (EV71) and coxsackievirus group A 16 (CVA16) are the main pathogens of human hand, foot and mouth disease. Among them, EV71 can cause severe infection of the central nervous system, and in severe cases, death of individuals. Compared with EV71, CVA16 causes milder clinical symptoms, but can also cause serious complications, such as pneumonia, encephalitis, and myocarditis (Pediatr Infect Dis J. 2004 Mar; 23(3):276-8.). EV71 is a single-stranded positive-strand RNA virus, belonging to the Picornaviridae family and enterovirus class A (Nat Struct Mol Biol. 19 (2012) 424-429).
EV71和CVA16均是无核膜包被的单股正链RNA病毒，为一个二十面体，直径大小为20-30nm，属于小核糖核酸病毒科，从属于肠道病毒。其全基因组大小约为7400bp，由5’非翻译区(UTR)和3’非翻译区(UTR)，以及一个开放阅读框(ORF)组成，编码一个多聚蛋白。开放阅读框可以分为三个亚级结构，称为P1、P2和P3区域。P1区域主要编码四个结构蛋白，分为VP1、VP2、VP3和VP4。四个结构蛋白组装形成一个原体，五个原体组成一个五聚体，12个五聚体一起形成病毒基因组的病毒体(Clinical and Experimental Vaccine Research.6(2017)4-14)。VP1、VP2和VP3在病毒衣壳的外面，可以作为病毒的抗原决定簇，VP4位于病毒衣壳的内部位置(Nature.317(1985)145-153)。P2区域编码三个非结构蛋白，分为2A、2B和2C。P3区域编码四个非结构蛋白，分为3A、3B、3C和3D。2A和3C作为EV71病毒的蛋白酶，可以切割病毒的多聚蛋白的成熟。3D蛋白是RNA聚合酶，在病毒翻译的过程中起到至关重要的作用(The Lancet Infectious Diseases.10(2010)778-790；Journal of Biomedical Science.21(2014)1-14)。在EV71 5’UTR
区域含有一个内部核糖体入核位点(IRES)，代替了真核生物的5’帽子结构，起始病毒的翻译过程。当EV71病毒颗粒进入宿主细胞后，释放病毒基因组，病毒RNA可以借助IRES-依赖的方式进行翻译(Journal of Virology.85(2011)9658-9666)。Both EV71 and CVA16 are single-stranded positive-sense RNA viruses without a nuclear membrane. They are an icosahedron with a diameter of 20-30nm. They belong to the Picornaviridae family and are subordinate to enteroviruses. Its full genome size is approximately 7400bp, consisting of a 5' untranslated region (UTR), a 3' untranslated region (UTR), and an open reading frame (ORF), encoding a polyprotein. The open reading frame can be divided into three sub-structures, called P1, P2 and P3 regions. The P1 region mainly encodes four structural proteins, divided into VP1, VP2, VP3 and VP4. Four structural proteins assemble to form a protomer, five protomers form a pentamer, and 12 pentamers together form the virion of the viral genome (Clinical and Experimental Vaccine Research. 6 (2017) 4-14). VP1, VP2 and VP3 are on the outside of the virus capsid and can serve as viral antigenic determinants, and VP4 is located on the inside of the virus capsid (Nature. 317 (1985) 145-153). The P2 region encodes three nonstructural proteins, divided into 2A, 2B and 2C. The P3 region encodes four nonstructural proteins, divided into 3A, 3B, 3C and 3D. 2A and 3C, as proteases of EV71 virus, can cleave the mature polyprotein of the virus. 3D protein is RNA polymerase and plays a vital role in the process of viral translation (The Lancet Infectious Diseases. 10 (2010) 778-790; Journal of Biomedical Science. 21 (2014) 1-14). The EV71 5'UTR This region contains an internal ribosome entry site (IRES), which replaces the eukaryotic 5' cap structure and initiates the viral translation process. When EV71 virus particles enter the host cell, the viral genome is released, and the viral RNA can be translated in an IRES-dependent manner (Journal of Virology. 85 (2011) 9658-9666).
EV71是一种高度的亲神经性的病毒，EV71感染最常见的靶点是脑干(The New England Journal of Medicine.341(1999)936-942)。EV71和CVA16感染途径与脊髓灰质炎病毒十分相似，主要有以下两个途径：第一种方式是病毒从血液经过血脑屏障，进而进入中枢神经系统，第二种方式是病毒通过末梢神经介导的逆向轴突运输进入中枢神经系统(Jama.207(1969)1481-1492；The Journal of General Virology.83(2002)1707-1720；Journal of Virology.78(2004)7186-7198；American Journal of Clinical Pathology.146(2016)95-106)。EV71能够导致脊髓灰质、脑干、下丘脑以及齿状核出现感染，并且炎症是其中最明显的感染症状(Journal of Neuropathology and Experimental Neurology.67(2008)162-169)。EV71 is a highly neurotropic virus, and the most common target of EV71 infection is the brainstem (The New England Journal of Medicine. 341 (1999) 936-942). The infection route of EV71 and CVA16 is very similar to that of poliovirus. There are two main ways: the first way is that the virus passes from the blood through the blood-brain barrier and then enters the central nervous system; the second way is that the virus is mediated through peripheral nerves. Reverse axonal transport into the central nervous system (Jama.207(1969)1481-1492; The Journal of General Virology.83(2002)1707-1720; Journal of Virology.78(2004)7186-7198; American Journal of Clinical Pathology .146(2016)95-106). EV71 can cause infection in the spinal cord gray matter, brainstem, hypothalamus and dentate nucleus, and inflammation is the most obvious infection symptom (Journal of Neuropathology and Experimental Neurology. 67 (2008) 162-169).
目前小分子化合物抑制EV71的方式主要有以下四种：1、抑制病毒进入宿主细胞。VP1的构象改变在病毒进入宿主的过程中至关重要，EV71的表面蛋白VP1是EV71抗原决定簇的中和性抗体主要识别位点。吡啶基咪唑烷酮化合物，Pleconari和BPR0Z-194，可以结合VP1抑制EV71病毒构象的改变，从而抑制EV71的复制(Arch Virol.157(2012)669-679；Antimicrob Agents Chemother.48(2004)3523-3529)。2、抑制EV71病毒的蛋白酶。EV71蛋白酶2A和3C可以切割EV71多聚蛋白的成熟，对于EV71的功能发挥至关重要。芦平曲韦(Rupintrivir)可以通过抑制EV71 3C蛋白酶的活性从而达到抑制EV71复制的效果(Journal of Virology.85(2011)10319-10331)。3、抑制EV71复制的关键蛋白。EV71RNA基因组复制依赖于EV71RNA聚合酶3D，所以靶向3D蛋白可以很好地抑制EV71复制。核苷酸类似物——病毒唑(ribavirin)和NITD008，以及非核苷酸类似物——DTriP-22，已经被证实可以抑制3D聚合酶(J Am Coll Cardiol.12(1988)1334-1341；Antimicrobial Agents and Chemotherapy.53(2009)2740-2747；Journal of Virology.88(2014)11915-11923)。4、抑制病毒的翻译过程。EV71RNA的翻译依赖于IRES，所以调节IRES的功能对于EV71的复制至关重要。山柰酚(一种黄酮类药物)已经被证实可以通过抑制EV71IRES的功能来抑制EV71的复制(Food Chem.128(2011)312-322)。At present, there are four main ways for small molecule compounds to inhibit EV71: 1. Inhibit the virus from entering host cells. The conformational change of VP1 is crucial in the process of virus entry into the host. The surface protein VP1 of EV71 is the main recognition site for neutralizing antibodies of EV71 epitopes. Pyridyl imidazolidinone compounds, Pleconari and BPR0Z-194, can bind to VP1 to inhibit the conformational changes of the EV71 virus, thereby inhibiting the replication of EV71 (Arch Virol. 157 (2012) 669-679; Antimicrob Agents Chemother. 48 (2004) 3523- 3529). 2. Inhibit the protease of EV71 virus. EV71 proteases 2A and 3C can cleave the maturation of EV71 polyprotein and are crucial for the function of EV71. Rupintrivir can inhibit EV71 replication by inhibiting the activity of EV71 3C protease (Journal of Virology. 85 (2011) 10319-10331). 3. Key proteins that inhibit EV71 replication. EV71 RNA genome replication depends on EV71 RNA polymerase 3D, so targeting the 3D protein can effectively inhibit EV71 replication. Nucleotide analogs, ribavirin and NITD008, and non-nucleotide analogs, DTriP-22, have been shown to inhibit 3D polymerase (J Am Coll Cardiol. 12 (1988) 1334-1341; Antimicrobial Agents and Chemotherapy.53(2009)2740-2747; Journal of Virology.88(2014)11915-11923). 4. Inhibit the translation process of the virus. The translation of EV71 RNA depends on IRES, so regulating the function of IRES is crucial for EV71 replication. Kaempferol (a flavonoid) has been shown to inhibit EV71 replication by inhibiting the function of the EV71 IRES (Food Chem. 128 (2011) 312-322).
EV71感染已经成为一个非常严重的公共安全问题，特别是在亚太地区。由于EV71引发的临床症状较CVA16严重，所以大多数科研团队更倾向于研发抗EV71的药物，而抗CVA16药物的研究较少。目前，临床上还没有治疗由EV71和/或CVA16感染引起的疾病
的特效药物，因此，开发有效地抗EV71和/或CVA16病毒的药物势在必行。EV71 infection has become a very serious public safety issue, especially in the Asia-Pacific region. Since the clinical symptoms caused by EV71 are more severe than those caused by CVA16, most scientific research teams prefer to develop anti-EV71 drugs, while there are fewer studies on anti-CVA16 drugs. Currently, there are no clinical treatments for disease caused by EV71 and/or CVA16 infection Therefore, it is imperative to develop drugs that are effective against EV71 and/or CVA16 viruses.
发明内容Contents of the invention
发明要解决的问题Invent the problem to be solved
本发明的目的是提供一种具有EV71和/或CVA16病毒抑制活性的化合物，及其在制备抗EV71和/或CVA16病毒药物中的应用。The object of the present invention is to provide a compound with EV71 and/or CVA16 virus inhibitory activity and its application in the preparation of anti-EV71 and/or CVA16 virus drugs.
用于解决问题的方案solutions to problems
本发明的目的通过以下技术方案得以实现：The purpose of the present invention is achieved through the following technical solutions:
<第一方面><First aspect>
本发明提供了一种具有EV71和/或CVA16病毒抑制活性的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体，所述化合物具有式(I)结构：
The present invention provides a compound with EV71 and/or CVA16 virus inhibitory activity or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof. The compound has the structure of formula (I):
The present invention provides a compound with EV71 and/or CVA16 virus inhibitory activity or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof. The compound has the structure of formula (I):
其中，in,
m为1-6中的任一整数；优选为1；m is any integer from 1 to 6; preferably 1;
n为0、1或2；优选为1；n is 0, 1 or 2; preferably 1;
R1各自独立地为氢、氘、卤素、C1-C6烷基、C1-C6卤代烷基或
R 1 is each independently hydrogen, deuterium, halogen, C1-C6 alkyl, C1-C6 haloalkyl or
若存在，R2各自独立地为氢、氘、卤素、C1-C6烷基或C1-C6卤代烷基；If present, R 2 is each independently hydrogen, deuterium, halogen, C1-C6 alkyl or C1-C6 haloalkyl;
R3为C1-C6烷基、C1-C6卤代烷基或C4-C6环烷基。R 3 is C1-C6 alkyl, C1-C6 haloalkyl or C4-C6 cycloalkyl.
优选地，所述化合物具有式(II)结构：
Preferably, the compound has the structure of formula (II):
Preferably, the compound has the structure of formula (II):
其中，R1、R2、R3如式(I)中所定义。
Among them, R 1 , R 2 and R 3 are as defined in formula (I).
优选地，R1为氢、氘、卤素或更优选氢、卤素或进一步优选氢、溴或
Preferably, R1 is hydrogen, deuterium, halogen or More preferably hydrogen, halogen or Further preference is given to hydrogen, bromine or
优选地，R2为C1-C6烷基，更优选C1-C4烷基，进一步优选甲基。Preferably, R 2 is C1-C6 alkyl, more preferably C1-C4 alkyl, further preferably methyl.
优选地，R3为C1-C6烷基，更优选C1-C4烷基，进一步优选异丙基。Preferably, R 3 is C1-C6 alkyl, more preferably C1-C4 alkyl, further preferably isopropyl.
进一步地，所述化合物为化合物1、化合物2或化合物3：
Further, the compound is compound 1, compound 2 or compound 3:
Further, the compound is compound 1, compound 2 or compound 3:
<第二方面><Second aspect>
本发明提供了一种药物组合物，其包含根据<第一方面>所述的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体。The present invention provides a pharmaceutical composition comprising the compound according to the <first aspect> or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof.
优选地，所述药物组合物还包含药学上可接受的辅料或载体。Preferably, the pharmaceutical composition further contains pharmaceutically acceptable excipients or carriers.
<第三方面><Third aspect>
本发明提供了一种药物制剂，其包含根据<第一方面>所述的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体，或者根据<第二方面>所述的药物组合物。The present invention provides a pharmaceutical preparation, which contains a compound according to the <first aspect> or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof, or a compound according to the <second aspect> >The pharmaceutical composition.
优选地，所述药物制剂为片剂、胶囊剂、注射剂、颗粒剂、粉剂、栓剂、丸剂、凝胶剂、散剂、口服溶液、吸入剂、混悬剂或干悬剂中的任意一种。
Preferably, the pharmaceutical preparation is any one of tablets, capsules, injections, granules, powders, suppositories, pills, gels, powders, oral solutions, inhalants, suspensions or dry suspensions.
<第四方面><The fourth aspect>
本发明提供了一种药物联合形式，其包含根据<第一方面>所述的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体，或者根据<第二方面>所述的药物组合物，或者根据<第三方面>所述的药物制剂，以及预防和/或治疗手足口病的药物中的一种或几种。The present invention provides a pharmaceutical combination form, which contains a compound according to the <first aspect> or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof, or a compound according to the <second aspect> Aspect> The pharmaceutical composition, or the pharmaceutical preparation according to <The third aspect>, and one or more of the drugs for preventing and/or treating hand, foot and mouth disease.
<第五方面><Fifth aspect>
本发明提供了根据<第一方面>所述的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体，或者根据<第二方面>所述的药物组合物，或者根据<第三方面>所述的药物制剂，或者根据<第四方面>所述的药物联合形式在制备抗EV71和/或CVA16病毒药物中的应用。The present invention provides a compound according to the <first aspect> or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof, or a pharmaceutical composition according to the <second aspect> , or according to the pharmaceutical preparation described in the <third aspect>, or according to the application of the pharmaceutical combination form described in the <fourth aspect> in the preparation of anti-EV71 and/or CVA16 viral drugs.
优选地，所述药物为预防和/或治疗至少部分应答于EV71和/或CVA16病毒的疾病和/或病症的药物。Preferably, the medicament is a medicament for the prevention and/or treatment of diseases and/or conditions that are at least partially responsive to EV71 and/or CVA16 viruses.
更优选地，所述药物为预防和/或治疗手足口病的药物。More preferably, the drug is a drug for preventing and/or treating hand, foot and mouth disease.
<第六方面><Sixth aspect>
本发明提供了根据<第一方面>所述的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体，或者根据<第二方面>所述的药物组合物，或者根据<第三方面>所述的药物制剂，或者根据<第四方面>所述的药物联合形式，其用于预防和/或治疗至少部分应答于EV71和/或CVA16病毒的疾病和/或病症。The present invention provides a compound according to the <first aspect> or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof, or a pharmaceutical composition according to the <second aspect> , or the pharmaceutical preparation according to the <third aspect>, or the pharmaceutical combination form according to the <fourth aspect>, which is used to prevent and/or treat diseases and/or at least partially responsive to EV71 and/or CVA16 viruses or illness.
优选地，所述至少部分应答于EV71和/或CVA16病毒的疾病和/或病症为手足口病。Preferably, the disease and/or condition that is at least partially responsive to EV71 and/or CVA16 viruses is hand, foot and mouth disease.
<第七方面><Seventh aspect>
本发明提供一种用于预防和/或治疗至少部分应答于EV71和/或CVA16病毒的疾病和/或病症的方法，其包括下列步骤：将预防和/或治疗有效量的根据<第一方面>所述的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体，或者根据<第二方面>所述的药物组合物，或者根据<第三方面>所述的药物制剂，或者根据<第四方面>所述的药物联合形式，施用于对其有需求的个体。The present invention provides a method for preventing and/or treating diseases and/or conditions that are at least partially responsive to EV71 and/or CVA16 viruses, which includes the following steps: adding a preventive and/or therapeutically effective amount of a drug according to <first aspect >The compound or its pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer, or the pharmaceutical composition according to the <second aspect>, or the pharmaceutical composition according to the <third aspect> The pharmaceutical preparation described above, or the pharmaceutical combination described in the <Fourth Aspect>, is administered to individuals in need thereof.
优选地，所述至少部分应答于EV71和/或CVA16病毒的疾病和/或病症为手足口病。Preferably, the disease and/or condition that is at least partially responsive to EV71 and/or CVA16 viruses is hand, foot and mouth disease.
发明的效果Effect of invention
本发明提供的化合物具有良好的EV71和/或CVA16病毒抑制活性，可以抑制EV71和/或CVA16病毒在宿主细胞内的复制及蛋白表达，且半数抑制浓度较低，有潜力作为抗EV71和/或CVA16病毒药物，用于临床上预防和/或治疗由EV71和/或CVA16病毒引起
的疾病和/或病症。The compound provided by the invention has good inhibitory activity against EV71 and/or CVA16 viruses, can inhibit the replication and protein expression of EV71 and/or CVA16 viruses in host cells, and has a low half inhibitory concentration, and has the potential to be used as an anti-EV71 and/or CVA16 virus. CVA16 virus drugs, used for clinical prevention and/or treatment caused by EV71 and/or CVA16 viruses diseases and/or conditions.
图1是实施例4中化合物1和2对EV71病毒滴度抑制的散点图。Figure 1 is a scatter plot showing the inhibition of EV71 virus titer by compounds 1 and 2 in Example 4.
图2是实施例4中化合物1和2抑制EV71病毒引起的细胞死亡的半数抑制率的柱形图。Figure 2 is a bar graph of the half-inhibition rate of compounds 1 and 2 in Example 4 in inhibiting cell death caused by EV71 virus.
图3是实施例4中化合物3抑制EV71病毒引起的细胞死亡的柱形图。Figure 3 is a bar graph showing that compound 3 inhibits cell death caused by EV71 virus in Example 4.
图4是实施例5中化合物1和2对EV71病毒RNA复制抑制的柱形图。Figure 4 is a bar graph showing the inhibition of EV71 viral RNA replication by compounds 1 and 2 in Example 5.
图5是实施例5中化合物3对EV71病毒RNA复制抑制的柱形图。Figure 5 is a bar graph showing the inhibition of EV71 viral RNA replication by Compound 3 in Example 5.
图6是实施例5中化合物1和2对EV71病毒RNA复制抑制的琼脂糖凝胶图。Figure 6 is an agarose gel image showing the inhibition of EV71 viral RNA replication by compounds 1 and 2 in Example 5.
图7是实施例6中化合物1和2对EV71病毒特异性3D表达抑制的免疫荧光图。Figure 7 is an immunofluorescence graph showing the inhibition of EV71 virus-specific 3D expression by Compounds 1 and 2 in Example 6.
图8是实施例6中化合物1和2不同浓度下对EV71病毒3D和VP1表达抑制的蛋白免疫印迹图。Figure 8 is a Western blot diagram showing the inhibition of EV71 virus 3D and VP1 expression by Compounds 1 and 2 at different concentrations in Example 6.
图9是实施例6中化合物3不同浓度下对EV71病毒3D和VP1表达抑制的蛋白免疫印迹图。Figure 9 is a Western blot diagram showing the inhibition of EV71 virus 3D and VP1 expression by Compound 3 at different concentrations in Example 6.
图10是实施例6中化合物1和2不同时间段对EV71病毒3D和VP1表达抑制的蛋白免疫印迹图。Figure 10 is a Western blot diagram showing the inhibition of EV71 virus 3D and VP1 expression by Compounds 1 and 2 at different time periods in Example 6.
图11是实施例7中化合物1抑制CVA16引起的细胞死亡的柱形图。Figure 11 is a bar graph showing that Compound 1 inhibits cell death caused by CVA16 in Example 7.
图12是实施例8中化合物1抑制CVA16病毒RNA复制的柱形图。Figure 12 is a bar graph of compound 1 inhibiting CVA16 viral RNA replication in Example 8.
图13是实施例9中化合物1抑制CVA16病毒3D蛋白表达的蛋白免疫印迹图。Figure 13 is a Western blot diagram showing that Compound 1 inhibits the expression of CVA16 virus 3D protein in Example 9.
为了对本发明的技术特征、目的和有益效果有更加清楚的理解，现对本发明的技术方案进行以下详细说明，但不能理解为对本发明的可实施范围的限定。In order to have a clearer understanding of the technical features, purposes and beneficial effects of the present invention, the technical solutions of the present invention are described in detail below, but this should not be understood as limiting the implementable scope of the present invention.
为了更为清晰地描述本发明的内容，现将所涉及的术语定义如下：In order to describe the content of the present invention more clearly, the terms involved are now defined as follows:
术语“药学上可接受的盐”表示本发明的化合物以它们的药用盐的形式存在，包括酸加成盐和碱加成盐。药学上可接受的盐在S.M.Berge在J.Pharmaceutical Sciences(66卷:1-19页,1977年)中描述的pharmaceutically salts中有所描述。在本发明中，药学上可接受的无毒的酸加成盐表示本发明中的化合物与有机或无机酸形成的盐，有机或无机酸包括(但不限于)盐酸、硫酸、氢溴酸、氢碘酸、磷酸、硝酸、高氯酸、乙酸、草酸、
马来酸、富马酸、酒石酸、苯磺酸、甲磺酸、水杨酸、琥珀酸、柠檬酸、乳酸、丙酸、苯甲酸、对甲苯磺酸和苹果酸等。药学上可接受的无毒的碱加成盐表示本发明中的化合物与有机或无机碱所形成的盐，包括(但不限于)碱金属盐，例如锂、钠或钾盐；碱土金属盐，例如钙或镁盐；有机碱盐，例如通过与含N基团的有机碱形成的铵盐或N+(C1-6烷基)4盐，优选为氢氧化锂、氢氧化钠、氢氧化钾、碳酸钠、碳酸氢钠、碳酸钾、碳酸氢钾、碳酸镁、碳酸钙、氨水、三乙胺或四丁基氢氧化铵等。“药学上可接受的盐”可通过一般的化学方法合成。The term "pharmaceutically acceptable salts" means that the compounds of the present invention exist in the form of their pharmaceutically acceptable salts, including acid addition salts and base addition salts. Pharmaceutically acceptable salts are described in Pharmaceutical Salts by SM Berge in J. Pharmaceutical Sciences (Vol. 66: pp. 1-19, 1977). In the present invention, pharmaceutically acceptable non-toxic acid addition salts refer to salts formed by compounds in the present invention and organic or inorganic acids, including (but not limited to) hydrochloric acid, sulfuric acid, hydrobromic acid, Hydroiodic acid, phosphoric acid, nitric acid, perchloric acid, acetic acid, oxalic acid, Maleic acid, fumaric acid, tartaric acid, benzenesulfonic acid, methanesulfonic acid, salicylic acid, succinic acid, citric acid, lactic acid, propionic acid, benzoic acid, p-toluenesulfonic acid and malic acid, etc. Pharmaceutically acceptable non-toxic base addition salts represent salts formed by compounds in the present invention and organic or inorganic bases, including (but not limited to) alkali metal salts, such as lithium, sodium or potassium salts; alkaline earth metal salts, For example, calcium or magnesium salts; organic base salts, such as ammonium salts or N + (C 1-6 alkyl) 4 salts formed with organic bases containing N groups, preferably lithium hydroxide, sodium hydroxide, hydroxide Potassium, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, magnesium carbonate, calcium carbonate, ammonia, triethylamine or tetrabutylammonium hydroxide, etc. "Pharmaceutically acceptable salts" can be synthesized by general chemical methods.
术语“溶剂化物”表示一个或多个溶剂分子与本发明中的化合物所形成的缔合物。形成溶剂化物的溶剂包括但不限于水、甲醇、乙醇、异丙醇、乙酸乙酯、四氢呋喃、N,N-二甲基甲酰胺、二甲亚砜等。The term "solvate" refers to an association of one or more solvent molecules with a compound of the present invention. Solvents that form solvates include, but are not limited to, water, methanol, ethanol, isopropyl alcohol, ethyl acetate, tetrahydrofuran, N,N-dimethylformamide, dimethyl sulfoxide, and the like.
术语“前药”表示作为本发明的化合物的化学衍生物，该衍生物在体内通过发生化学反应转换成本发明的化合物。The term "prodrug" refers to a chemical derivative of a compound of the present invention that is converted into a compound of the present invention by a chemical reaction in the body.
术语“同位素标记物”表示同位素包括(但不只限于)2H、3H、11C、13C、14C、15N等。The term "isotope label" means isotopes including (but not limited to) 2 H, 3 H, 11 C, 13 C, 14 C, 15 N, etc.
术语“异构体”包含所有的同分异构形式，包括对映异构体、非对映异构体、互变异构体和几何异构体(包括顺反异构体)。因此，本发明中所设计的化合物的单个立体化学异构体或其对映异构体、非对映异构体、互变异构体或几何异构体(或顺反异构体)的混合物都属于本发明的范围。The term "isomers" includes all isomeric forms, including enantiomers, diastereomers, tautomers and geometric isomers (including cis and trans isomers). Therefore, the individual stereochemical isomers or enantiomers, diastereomers, tautomers or geometric isomers (or cis-trans isomers) of the compounds designed in the present invention Mixtures are within the scope of the invention.
术语“预防”是指例如当患者或受试者处于易患病状或者处于疾病或病状的风险中时，可以完全或几乎完全阻止疾病或病状(例如感染、缺血或再灌注损伤)发生；预防还可以包括抑制，即阻止病状的发展。The term "prevention" means, for example, the complete or almost complete prevention of the occurrence of a disease or condition (e.g., infection, ischemia or reperfusion injury) when a patient or subject is susceptible to the condition or at risk of the disease or condition; Prevention can also include suppression, that is, halting the progression of the condition.
术语“治疗”是指：1)抑制该疾病；例如，抑制正在经历或显示疾病、病状或病症的病理学或症状学的个体的该疾病、病状或病症(即，阻止病理学和/或症状学的进一步发展)；或2)改善该疾病；例如，改善正在经历或显示疾病、病状或病症的病理学或症状学的个体的该疾病、病状或病症(即，逆转病理学和/或症状学)。The term "treating" means: 1) inhibiting the disease; e.g., inhibiting the disease, condition, or condition in an individual who is experiencing or exhibiting pathology or symptomatology of the disease, condition, or condition (i.e., preventing the pathology and/or symptoms or 2) ameliorating the disease; e.g., ameliorating the disease, condition, or condition in an individual who is experiencing or exhibiting the pathology or symptomatology of the disease, condition, or disorder (i.e., reversing the pathology and/or symptoms study).
下述实施例中所述实验方法，如无特殊说明，均为常规方法；所述试剂和材料，如无特殊说明，均可从商业途径获得。The experimental methods described in the following examples are conventional methods unless otherwise specified; the reagents and materials described can be obtained from commercial sources unless otherwise specified.
在下述的实施例中，所用溶剂和药品均为分析纯或化学纯；溶剂在使用前均经过重新蒸馏；无水溶剂均按照标准方法或文献方法进行处理。In the following examples, all solvents and drugs used are of analytical or chemical purity; all solvents are redistilled before use; all anhydrous solvents are processed according to standard methods or literature methods.
柱层析硅胶(100-200目)和薄层层析硅胶(GF254)为青岛海洋化工厂和烟台化工
厂产品；如未特别说明，均采用石油醚(60-90℃)/乙酸乙酯(v/v)作为洗脱剂。Column chromatography silica gel (100-200 mesh) and thin layer chromatography silica gel (GF254) are from Qingdao Ocean Chemical Plant and Yantai Chemical Industry Factory products; unless otherwise specified, petroleum ether (60-90°C)/ethyl acetate (v/v) is used as the eluent.
所有萃取溶剂未经说明，均用无水Na2SO4干燥。All extraction solvents were dried over anhydrous Na2SO4 without being stated.
1H NMR用varian-400型核磁共振仪记录，TMS(四甲基硅烷)为内标。 1 H NMR was recorded with a varian-400 nuclear magnetic resonance instrument, and TMS (tetramethylsilane) was used as the internal standard.
在活性效果测试中，若无特别说明，溶解化合物的溶剂均采用DMSO(二甲亚砜)。In the activity effect test, unless otherwise specified, DMSO (dimethyl sulfoxide) was used as the solvent to dissolve the compounds.
实施例1：Example 1:
化合物1是由如下方法合成的：
Compound 1 was synthesized by the following method:
Compound 1 was synthesized by the following method:
将市售化合物1-1(160mg,1.1mmol)和2,4-二氯-6-甲基嘧啶(200mg,1.2mmol)溶于N-甲基吡咯烷酮(15mL)中。再向反应体系加入N,N-二异丙基乙胺(194mg,1.5mmol)后，于130℃下搅拌过夜。TLC板检测反应完全后，反应液冷至室温。向反应液中依次加入异丙胺(729mg,12mmol)、N,N-二异丙基乙胺(194mg,1.5mmol)后，于120℃下搅拌过夜。反应液冷至室温后，加入乙酸乙酯稀释(50mL)，有机相用饱和食盐水洗涤(20mL*5)，有机相分离后用无水硫酸钠干燥，减压浓缩。残留物用硅胶柱层析纯化(二氯甲烷/甲醇＝100/1-20/1)得黄色固体化合物1(20mg,6.2％)。Commercial compound 1-1 (160 mg, 1.1 mmol) and 2,4-dichloro-6-methylpyrimidine (200 mg, 1.2 mmol) were dissolved in N-methylpyrrolidone (15 mL). After adding N,N-diisopropylethylamine (194 mg, 1.5 mmol) to the reaction system, the mixture was stirred at 130°C overnight. After the TLC plate detects that the reaction is complete, the reaction solution is cooled to room temperature. Isopropylamine (729 mg, 12 mmol) and N,N-diisopropylethylamine (194 mg, 1.5 mmol) were added in sequence to the reaction solution, and the mixture was stirred at 120° C. overnight. After the reaction solution was cooled to room temperature, ethyl acetate was added to dilute (50 mL). The organic phase was washed with saturated brine (20 mL*5). After separation, the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane/methanol=100/1-20/1) to obtain compound 1 as a yellow solid (20 mg, 6.2%).
1H NMR(400MHz,CDCl3)δ8.84(s,1H),8.14(s,1H),8.11-8.02(m,2H),7.68-7.59(m,1H),7.40(s,1H),7.26(s,1H),7.09(s,1H),5.91(s,1H),4.24-4.06(m,1H),2.25(s,3H),1.30(s,6H). 1 H NMR (400MHz, CDCl 3 ) δ8.84(s,1H),8.14(s,1H),8.11-8.02(m,2H),7.68-7.59(m,1H),7.40(s,1H), 7.26(s,1H),7.09(s,1H),5.91(s,1H),4.24-4.06(m,1H),2.25(s,3H),1.30(s,6H).
实施例2：Example 2:
化合物2是由如下方法合成的：
Compound 2 was synthesized by the following method:
Compound 2 was synthesized by the following method:
1)中间体2-2的合成：
1) Synthesis of intermediate 2-2:
将4-氯-6-甲基-2-甲硫基嘧啶(524mg,3.0mmol)与化合物2-1(669mg,3.0mmol)溶于无水四氢呋喃(5mL)中。氮气环境下，在冰水浴下逐滴加入二(三甲基硅基)氨基钠(3.0mL,2M,6.0mmol)的四氢呋喃溶液。常温反应两小时后，饱和碳酸氢钠水溶液(5mL)加入淬灭反应，用乙酸乙酯(10mL)萃取三次。有机相合并，干燥浓缩。残余物经柱层析纯化(乙酸乙酯/石油醚＝1/1-1/0)得中间体2-2(758mg,70％)，为淡黄色固体。4-Chloro-6-methyl-2-methylthiopyrimidine (524 mg, 3.0 mmol) and compound 2-1 (669 mg, 3.0 mmol) were dissolved in anhydrous tetrahydrofuran (5 mL). Under a nitrogen atmosphere, a tetrahydrofuran solution of sodium bis(trimethylsilyl)amide (3.0 mL, 2 M, 6.0 mmol) was added dropwise in an ice-water bath. After reacting at room temperature for two hours, saturated aqueous sodium bicarbonate solution (5 mL) was added to quench the reaction, and the mixture was extracted three times with ethyl acetate (10 mL). The organic phases were combined, dried and concentrated. The residue was purified by column chromatography (ethyl acetate/petroleum ether=1/1-1/0) to obtain intermediate 2-2 (758 mg, 70%) as a light yellow solid.
1H NMR(400MHz,CDCl3)δ8.81(d,J＝2.4Hz,1H),8.23(d,J＝2.0Hz,1H),8.04(d,J＝9.2Hz,1H),8.01(d,J＝2.4Hz,1H),7.64-7.62(m,1H),6.89(s,1H),6.31(s,1H),2.60(s,3H),2.36(s,3H). 1 H NMR (400MHz, CDCl 3 ) δ8.81(d,J＝2.4Hz,1H),8.23(d,J＝2.0Hz,1H),8.04(d,J＝9.2Hz,1H),8.01(d ,J＝2.4Hz,1H),7.64-7.62(m,1H),6.89(s,1H),6.31(s,1H),2.60(s,3H),2.36(s,3H).
2)中间体2-3的合成：2) Synthesis of intermediate 2-3:
将中间体2-2(361mg,1.0mmol)溶于(3mL)四氢呋喃中，使用氮气保护后，加入水(0.15mL)与单过硫酸氢钾(636mg,2.0mmol)，室温搅拌过夜。减压浓缩，得到粗品中间体2-3(1.0g)，为黄色固体，直接用于下一步反应。Intermediate 2-2 (361 mg, 1.0 mmol) was dissolved in (3 mL) tetrahydrofuran. After using nitrogen protection, water (0.15 mL) and potassium monopersulfate (636 mg, 2.0 mmol) were added, and the mixture was stirred at room temperature overnight. Concentrate under reduced pressure to obtain crude intermediate 2-3 (1.0g) as a yellow solid, which was directly used in the next reaction.
3)化合物2的合成：3) Synthesis of compound 2:
将粗品中间体2-3(1.0g,1.0mmol)溶于N-甲基吡咯烷酮(5mL)中，加入N,N-二异丙基乙基胺(645mg,5.0mmol)和异丙胺(590mg,10.0mmol)，100℃下搅拌过夜。待冷却至常温，将溶液倒入冰水(20mL)中，有固体析出。过滤，固体使用乙醚淋洗，经柱层析纯化(乙酸乙酯/石油醚＝1/1-1/0)得到黄色固体化合物2(223mg,60％)。Dissolve crude intermediate 2-3 (1.0g, 1.0mmol) in N-methylpyrrolidone (5mL), add N,N-diisopropylethylamine (645mg, 5.0mmol) and isopropylamine (590mg, 10.0 mmol), stir at 100°C overnight. After cooling to room temperature, the solution was poured into ice water (20 mL), and a solid precipitated. The solid was filtered, rinsed with diethyl ether, and purified by column chromatography (ethyl acetate/petroleum ether = 1/1-1/0) to obtain compound 2 as a yellow solid (223 mg, 60%).
1H NMR(400MHz,CDCl3)δ8.78(d,J＝1.6Hz,1H),8.20(s,1H),8.11(s,1H),8.00(d,J＝9.2Hz,1H),7.62(dd,J＝9.2,1.6Hz,1H),6.66(s,1H),5.95(s,1H),4.86(d,J＝6.4Hz,1H),4.25-4.12(m,1H),2.25(s,3H),1.29(d,J＝5.6Hz,6H). 1 H NMR (400MHz, CDCl 3 ) δ8.78 (d, J = 1.6 Hz, 1H), 8.20 (s, 1H), 8.11 (s, 1H), 8.00 (d, J = 9.2 Hz, 1H), 7.62 (dd,J＝9.2,1.6Hz,1H),6.66(s,1H),5.95(s,1H),4.86(d,J＝6.4Hz,1H),4.25-4.12(m,1H),2.25( s,3H),1.29(d,J＝5.6Hz,6H).
实施例3：Example 3:
化合物3是由如下方法合成的：
Compound 3 was synthesized by the following method:
Compound 3 was synthesized by the following method:
1)中间体3-1的合成：1) Synthesis of intermediate 3-1:
将化合物2(100mg,0.27mmol)溶于N-甲基吡咯烷酮(2mL)，加入炔丙基氨基甲酸叔丁酯(93mg,0.6mmol)、二异丙胺(1mL)、二(三苯基磷)二氯化钯(7mg,0.01mmol)和CuI(1mg,0.005mmol)，在氮气气氛下加热到100℃，搅拌20h。待冷却至常温，滤除不溶固体。滤液倒入10mL水中，乙酸乙酯萃取3次。有机相合并，用饱和食盐水洗涤3次。有机相经无水硫酸钠干燥，减压浓缩。残余物经柱层析纯化(乙酸乙酯/石油醚＝1/1-1/0)得黄色固体中间体3-1(80mg,66％)。Compound 2 (100 mg, 0.27 mmol) was dissolved in N-methylpyrrolidone (2 mL), and tert-butyl propargyl carbamate (93 mg, 0.6 mmol), diisopropylamine (1 mL), and bis(triphenylphosphorus) were added. Palladium dichloride (7 mg, 0.01 mmol) and CuI (1 mg, 0.005 mmol) were heated to 100°C under a nitrogen atmosphere and stirred for 20 h. After cooling to room temperature, filter out the insoluble solid. The filtrate was poured into 10 mL of water and extracted with ethyl acetate three times. The organic phases were combined and washed three times with saturated brine. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (ethyl acetate/petroleum ether=1/1-1/0) to obtain yellow solid intermediate 3-1 (80 mg, 66%).
2)中间体3-2的合成：2) Synthesis of intermediate 3-2:
将中间体3-1(80mg,0.18mmol)溶于甲醇(2mL)，加入10％钯碳(8mg)。置换氢气，常温反应过夜。反应液减压浓缩，得到黄色固体中间体3-2(80mg,100％)，直接用于下一步反应。Intermediate 3-1 (80 mg, 0.18 mmol) was dissolved in methanol (2 mL), and 10% palladium on carbon (8 mg) was added. Replace hydrogen gas and react overnight at room temperature. The reaction solution was concentrated under reduced pressure to obtain yellow solid intermediate 3-2 (80 mg, 100%), which was directly used in the next reaction.
1H NMR(400MHz,CDCl3)δ8.70(s,1H),8.04(d,J＝9.2Hz,1H),7.98(s,1H),7.83(s,1H),7.59(d,J＝8.8Hz,1H),7.05-6.98(m,1H),5.89(s,1H),4.65-4.57(m,1H),4.20-4.10(m,1H),3.38(t,J＝6.8Hz,2H),3.27-3.17(m,2H),2.09(s,3H),2.05-2.00(m,1H),1.95-1.90(m,1H),1.45(s,9H),1.30(d,J＝6.8Hz,6H). 1 H NMR (400MHz, CDCl 3 ) δ8.70 (s, 1H), 8.04 (d, J = 9.2 Hz, 1H), 7.98 (s, 1H), 7.83 (s, 1H), 7.59 (d, J = 8.8Hz,1H),7.05-6.98(m,1H),5.89(s,1H),4.65-4.57(m,1H),4.20-4.10(m,1H),3.38(t,J＝6.8Hz,2H ),3.27-3.17(m,2H),2.09(s,3H),2.05-2.00(m,1H),1.95-1.90(m,1H),1.45(s,9H),1.30(d,J＝6.8 Hz,6H).
3)中间体3-3的合成：3) Synthesis of intermediate 3-3:
中间体3-2(80mg,0.18mmol)溶于甲醇(1mL)，加入3M氯化氢的甲醇溶液(1mL,3mmol)，常温搅拌5h。待反应完毕，减压浓缩得黄色固体中间体3-3(70mg,100％)，为盐酸盐，直接用于下一步反应。Intermediate 3-2 (80 mg, 0.18 mmol) was dissolved in methanol (1 mL), 3M hydrogen chloride in methanol solution (1 mL, 3 mmol) was added, and stirred at room temperature for 5 h. After the reaction is completed, concentrate under reduced pressure to obtain yellow solid intermediate 3-3 (70 mg, 100%) as hydrochloride, which can be directly used in the next step of the reaction.
4)化合物3的合成：4) Synthesis of compound 3:
中间体3-3(70mg,0.18mmol)溶于N,N-二甲基甲酰胺(1mL)，加入化合物
Biotin-NC-3(123mg,0.27mmol)和三乙胺(50mg,0.5mmol)，常温搅拌过夜。待反应完毕，减压浓缩，经柱层析纯化(CH2Cl2/MeOH/NH4OH＝100/10/1)得黄色固体化合物3(28mg,22％)。Intermediate 3-3 (70 mg, 0.18 mmol) was dissolved in N,N-dimethylformamide (1 mL), and the compound was added Biotin-NC-3 (123 mg, 0.27 mmol) and triethylamine (50 mg, 0.5 mmol) were stirred at room temperature overnight. After the reaction was completed, the reaction mixture was concentrated under reduced pressure and purified by column chromatography (CH 2 Cl 2 /MeOH/NH 4 OH = 100/10/1) to obtain compound 3 (28 mg, 22%) as a yellow solid.
1H NMR(400MHz,DMSO-d6)δ9.44(br s,1H),8.59(s,1H),8.56(br s,1H),7.94(br s,1H),7.86(d,J＝8.8Hz,1H),7.85(s,1H),7.78(d,J＝8.8Hz,1H),7.75-7.70(m,1H),8.68(br s,1H),6.42(s,1H),6.36(s,1H),5.96(s,1H),4.32-4.26(m,1H),4.16-4.06(m,2H),3.15-3.05(m,3H),3.04-2.97(m,2H),2.84-2.73(m,3H),2.57(d,J＝12.4Hz,1H),2.15(s,3H),2.10-1.98(m,4H),1.85-1.75(m,2H),1.65-1.55(m,1H),1.53-1.43(m,5H),1.42-1.33(m,2H),1.30-1.18(m,10H). 1 H NMR (400MHz, DMSO-d 6 ) δ9.44 (br s, 1H), 8.59 (s, 1H), 8.56 (br s, 1H), 7.94 (br s, 1H), 7.86 (d, J＝ 8.8Hz,1H),7.85(s,1H),7.78(d,J＝8.8Hz,1H),7.75-7.70(m,1H),8.68(br s,1H),6.42(s,1H),6.36 (s,1H),5.96(s,1H),4.32-4.26(m,1H),4.16-4.06(m,2H),3.15-3.05(m,3H),3.04-2.97(m,2H),2.84 -2.73(m,3H),2.57(d,J＝12.4Hz,1H),2.15(s,3H),2.10-1.98(m,4H),1.85-1.75(m,2H),1.65-1.55(m ,1H),1.53-1.43(m,5H),1.42-1.33(m,2H),1.30-1.18(m,10H).
实施例4：化合物1、2和3的抗EV71病毒活性测试Example 4: Anti-EV71 virus activity test of compounds 1, 2 and 3
实验步骤：Experimental steps:
1)将RD细胞种在12板孔中，隔天待细胞长至90％，加入MOI＝0.25的EV71感染RD细胞；1) Seed RD cells in 12 plate wells, wait until the cells grow to 90% the next day, then add EV71 with MOI=0.25 to infect the RD cells;
2)24小时后，用细胞刮刀将细胞刮下，收取细胞碎片和病毒上清，-80℃反复冻融三次，2000转速/5分钟，收取上清，将收取的病毒做10倍梯度稀释，每个浓度做3个复孔；2) After 24 hours, use a cell scraper to scrape off the cells, collect the cell fragments and virus supernatant, freeze and thaw repeatedly at -80°C three times, 2000 rpm/5 minutes, collect the supernatant, and make a 10-fold gradient dilution of the collected virus. Make 3 duplicate wells for each concentration;
3)在96孔板中种RD细胞，待细胞生长90％，弃去培养基，加入梯度稀释的病毒的同时加入不同梯度的化合物1、2或3(5μM、10μM和20μM)处理RD细胞；3) Seed RD cells in a 96-well plate. When the cells grow 90%, discard the medium, add gradient dilutions of virus and add different gradients of compound 1, 2 or 3 (5 μM, 10 μM and 20 μM) to treat the RD cells;
4)24小时后，弃去培养基，PBS洗一遍，每孔加入30μL结晶紫，孵育2分钟后，用双蒸水反复清洗，干燥后，在显微镜下数噬菌斑，计算滴度。4) After 24 hours, discard the medium, wash once with PBS, add 30 μL of crystal violet to each well, incubate for 2 minutes, wash repeatedly with double-distilled water, dry, count plaques under a microscope, and calculate the titer.
通过图1和图2得知，随着浓度梯度的升高，化合物1和2可以有效地抑制EV71在RD细胞中的病毒滴度(图1)。并且，进一步发现，化合物1和2抑制EV71引起RD细胞死亡的半数抑制浓度(IC50)分别为3.075μM和5.716μM(图2)。化合物3在10μM和20μM都能显著抑制EV71引起的RD细胞死亡，一定范围内呈现出剂量依赖性(图3)。图3中的“D”表示只预处理溶媒DMSO，未用化合物3预处理RD细胞。It can be seen from Figures 1 and 2 that as the concentration gradient increases, compounds 1 and 2 can effectively inhibit the viral titer of EV71 in RD cells (Figure 1). Moreover, it was further found that the half inhibitory concentrations (IC 50 ) of compounds 1 and 2 that inhibited EV71-induced RD cell death were 3.075 μM and 5.716 μM, respectively (Figure 2). Compound 3 can significantly inhibit EV71-induced RD cell death at both 10 μM and 20 μM, showing a dose-dependent manner within a certain range (Figure 3). “D” in Figure 3 indicates that only vehicle DMSO was pretreated, and RD cells were not pretreated with compound 3.
实施例5：化合物1、2和3对EV71病毒复制的抑制能力测试Example 5: Test of the inhibitory ability of compounds 1, 2 and 3 on EV71 virus replication
实验步骤：Experimental steps:
1)分别用化合物1、2和3预处理RD细胞1小时；1) Pretreat RD cells with compounds 1, 2 and 3 respectively for 1 hour;
2)用EV71感染上述RD细胞8小时；2) Infect the above RD cells with EV71 for 8 hours;
3)弃去上清，收细胞，提取RNA，反转成cDNA；
3) Discard the supernatant, collect the cells, extract RNA, and reverse it into cDNA;
4)利用EV71的特异性QPCR引物，检测EV71病毒RNA在细胞中的表达量，利用半定量PCR的方法定量EV71的RNA水平。4) Use EV71-specific QPCR primers to detect the expression of EV71 viral RNA in cells, and use semi-quantitative PCR to quantify EV71 RNA levels.
通过图4、图5和图6得知，化合物1、2和3可以有效地减少EV71RNA在细胞内的复制(图4和图5)。同时，采用半定量PCR的方法定量EV71的RNA水平，检测EV71的特异性RNA片段：VP1、2A、2AB、3ABC和3D，发现化合物1和2可以有效地抑制EV71RNA的复制(图6)。图4、图5以及图6中的“D”表示只处理溶媒DMSO，未用化合物1、2和3预处理RD细胞。It can be seen from Figure 4, Figure 5 and Figure 6 that compounds 1, 2 and 3 can effectively reduce the replication of EV71 RNA in cells (Figure 4 and Figure 5). At the same time, semi-quantitative PCR was used to quantify the RNA level of EV71 and detect the specific RNA fragments of EV71: VP1, 2A, 2AB, 3ABC and 3D. It was found that compounds 1 and 2 can effectively inhibit the replication of EV71 RNA (Figure 6). “D” in Figure 4, Figure 5 and Figure 6 indicates that only vehicle DMSO was treated, and RD cells were not pretreated with compounds 1, 2 and 3.
实施例6：化合物1、2和3对EV71病毒蛋白表达的抑制能力测试Example 6: Test of the inhibitory ability of compounds 1, 2 and 3 on EV71 viral protein expression
免疫荧光实验步骤：Immunofluorescence experimental steps:
1)将RD细胞种在置于6孔板的玻璃片种，待细胞生长至80％，分别用化合物1和2预处理RD细胞1小时；1) Seed RD cells on a glass plate placed in a 6-well plate. When the cells grow to 80%, pretreat the RD cells with compounds 1 and 2 respectively for 1 hour;
2)加入MOI＝0.25的EV71感染RD细胞；2) Add EV71 with MOI=0.25 to infect RD cells;
3)8小时后，弃去培养基，再加入PBS清洗一遍，用4％多聚甲醛固定细胞10分钟，再加入PBS清洗三次；3) After 8 hours, discard the culture medium, add PBS and wash once, fix the cells with 4% paraformaldehyde for 10 minutes, then add PBS and wash three times;
4)用0.3％Triton X-100渗透细胞，再用3％BSA封闭细胞1.5小时；加入1:1000稀释的EV71-3D抗体过夜，PBS清洗三次，加入1:1000稀释的荧光二抗1.5小时，并且避光操作；4) Permeabilize the cells with 0.3% Triton And operate away from light;
5)最后，在避光条件下用DAPI染细胞核10分钟，利用荧光显微镜拍片，检查病毒3D蛋白的表达。5) Finally, stain the cell nuclei with DAPI for 10 minutes in the dark, and use a fluorescence microscope to take pictures to check the expression of the viral 3D protein.
Western blot实验步骤：Western blot experimental steps:
1)将RD细胞种于6孔板中，密度为5*105个/每孔；1) Seed RD cells in a 6-well plate at a density of 5*10 5 cells/well;
2)24小时后，用不同浓度的化合物1、2或3预处理RD细胞1小时；2) After 24 hours, RD cells were pretreated with different concentrations of compound 1, 2 or 3 for 1 hour;
3)用EV71感染RD细胞8小时；3) Infect RD cells with EV71 for 8 hours;
4)弃去上清，收集细胞，应用细胞裂解液重悬细胞，并置于冰上20min；4) Discard the supernatant, collect the cells, resuspend the cells in cell lysis buffer, and place on ice for 20 minutes;
5)4℃，13000×g离心20min后收集上清，western blot分析目的条带。5) Collect the supernatant after centrifugation at 13000×g for 20 minutes at 4°C, and analyze the target band by western blot.
实验结果如图7至图10所示。The experimental results are shown in Figures 7 to 10.
通过免疫荧光实验，我们发现化合物1和2可以有效地抑制EV71特异性蛋白3D在宿主细胞内的表达(图7)。在相同的时间条件下，我们用不同浓度的化合物1、2和3处理RD细胞1个小时，再加入EV71感染RD细胞8小时，发现随着浓度梯度的升高，化合物1、2和3可以有效地抑制EV71特异性蛋白VP1和3D的表达(图8和图9)。用
相同浓度的化合物1和2处理RD细胞，在EV71感染RD细胞的不同时间条件下，8小时、10小时以及12小时，EV71病毒特异性蛋白的表达随着时间的延长越来越多，而化合物1和2在不同的时间条件下都可以有效地抑制病毒特异性蛋白的表达(见图10，图10中的“D”表示只预处理溶媒DMSO，未用化合物1和2预处理RD细胞)。Through immunofluorescence experiments, we found that compounds 1 and 2 can effectively inhibit the expression of EV71-specific protein 3D in host cells (Figure 7). Under the same time conditions, we treated RD cells with different concentrations of compounds 1, 2 and 3 for 1 hour, and then added EV71 to infect RD cells for 8 hours. We found that as the concentration gradient increased, compounds 1, 2 and 3 could Effectively inhibited the expression of EV71-specific proteins VP1 and 3D (Figure 8 and Figure 9). use The same concentration of compounds 1 and 2 treated RD cells. Under different time conditions of EV71 infection of RD cells, 8 hours, 10 hours and 12 hours, the expression of EV71 virus-specific proteins increased with time, while the expression of compounds 1 and 2 can effectively inhibit the expression of virus-specific proteins under different time conditions (see Figure 10, "D" in Figure 10 indicates that only the vehicle DMSO was pretreated, and RD cells were not pretreated with compounds 1 and 2) .
综合实施例4-6，我们可以得出结论：小分子化合物1、2和3可以有效地抑制EV71在宿主细胞内的复制。Based on Examples 4-6, we can conclude that small molecule compounds 1, 2 and 3 can effectively inhibit the replication of EV71 in host cells.
实施例7：化合物1对CVA16病毒引起的细胞死亡的抑制能力测试Example 7: Test of the ability of compound 1 to inhibit cell death caused by CVA16 virus
实验步骤：Experimental steps:
1)将RD细胞种于96孔板中，密度为5000个/每孔；1) Seed RD cells in a 96-well plate at a density of 5,000 cells/well;
2)24小时后，用化合物1预处理RD细胞2h，化合物的浓度为20μM；2) After 24 hours, pretreat RD cells with compound 1 for 2 hours, the concentration of the compound is 20 μM;
3)用CVA16感染RD细胞18小时；3) Infect RD cells with CVA16 for 18 hours;
4)应用Cell Titer-Glo Luminescent Cell Viability Assay kit检测细胞活性。4) Use Cell Titer-Glo Luminescent Cell Viability Assay kit to detect cell viability.
实验结果如图11所示，化合物1可以有效地减少CVA16引起的RD细胞死亡。The experimental results are shown in Figure 11. Compound 1 can effectively reduce RD cell death caused by CVA16.
实施例8：化合物1对CVA16病毒RNA复制的抑制能力测试Example 8: Test of the ability of compound 1 to inhibit CVA16 viral RNA replication
实验步骤：Experimental steps:
1)将RD细胞种于6孔板中，密度为3*105个/每孔；1) Seed RD cells in a 6-well plate at a density of 3*10 5 cells/well;
2)24小时后，用不同浓度的化合物1预处理RD细胞2h；2) After 24 hours, RD cells were pretreated with different concentrations of compound 1 for 2 hours;
3)用CVA16(MOI＝5)感染RD细胞8h；3) Infect RD cells with CVA16 (MOI=5) for 8 hours;
4)弃去上清，收集细胞，提取总RNA并反转成cDNA；4) Discard the supernatant, collect cells, extract total RNA and reverse it into cDNA;
5)利用CVA16的特异性QPCR引物，检测CVA16病毒RNA在细胞中的表达量。5) Use CVA16-specific QPCR primers to detect the expression of CVA16 viral RNA in cells.
实验结果如图12所示，化合物1可以有效地减少CVA16病毒RNA的复制，并且在一定范围内呈现出剂量依赖性。The experimental results are shown in Figure 12. Compound 1 can effectively reduce the replication of CVA16 viral RNA in a dose-dependent manner within a certain range.
实施例9：化合物1对CVA16病毒蛋白表达的抑制能力测试Example 9: Test of the ability of compound 1 to inhibit the expression of CVA16 viral protein
实验步骤：Experimental steps:
1)将RD细胞种于6孔板中，密度为5*105个/每孔；1) Seed RD cells in a 6-well plate at a density of 5*10 5 cells/well;
2)24小时后，用不同浓度的化合物1预处理RD细胞2h；2) After 24 hours, RD cells were pretreated with different concentrations of compound 1 for 2 hours;
3)用CVA16(MOI＝5)感染RD细胞8h；3) Infect RD cells with CVA16 (MOI=5) for 8 hours;
4)弃去上清，收集细胞，应用细胞裂解液重悬细胞，并置于冰上20min；4) Discard the supernatant, collect the cells, resuspend the cells in cell lysis buffer, and place on ice for 20 minutes;
5)4℃，13000×g离心20min后收集上清，western blot分析目的条带。5) Collect the supernatant after centrifugation at 13000 × g for 20 minutes at 4°C and analyze the target band by western blot.
实验结果如图13所示，化合物1可以有效地抑制CVA16病毒的3D蛋白的表达(图
13)。综合图11-13，我们可以得出小分子化合物1可以有效地抑制CVA16在宿主细胞内的复制。The experimental results are shown in Figure 13. Compound 1 can effectively inhibit the expression of the 3D protein of CVA16 virus (Figure 13). Based on Figures 11-13, we can conclude that small molecule compound 1 can effectively inhibit the replication of CVA16 in host cells.
综上，本发明的化合物可以有效地抑制EV71和/或CVA16在宿主细胞内的复制，有潜力作为临床上预防和/或治疗由EV71和/或CVA16引起的疾病和/或病症的药物。
In summary, the compounds of the present invention can effectively inhibit the replication of EV71 and/or CVA16 in host cells, and have the potential to be used as clinical drugs for preventing and/or treating diseases and/or conditions caused by EV71 and/or CVA16.
Claims (9)
- 具有式(I)结构的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体，
A compound having the structure of formula (I) or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof,
其中，in,m为1-6中的任一整数；m is any integer from 1 to 6;n为0、1或2；n is 0, 1 or 2;R1各自独立地为氢、氘、卤素、C1-C6烷基、C1-C6卤代烷基或 R 1 is each independently hydrogen, deuterium, halogen, C1-C6 alkyl, C1-C6 haloalkyl or若存在，R2各自独立地为氢、氘、卤素、C1-C6烷基或C1-C6卤代烷基；If present, R 2 is each independently hydrogen, deuterium, halogen, C1-C6 alkyl or C1-C6 haloalkyl;R3为C1-C6烷基、C1-C6卤代烷基或C4-C6环烷基。R 3 is C1-C6 alkyl, C1-C6 haloalkyl or C4-C6 cycloalkyl. - 根据权利要求1所述的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体，其特征在于，The compound according to claim 1 or its pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer, characterized in that,m为1；n为1。m is 1; n is 1.
- 根据权利要求1或2所述的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体，其特征在于，所述化合物具有式(II)结构，
The compound according to claim 1 or 2 or its pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer, characterized in that the compound has the structure of formula (II),
其中，R1、R2、R3如权利要求1中所定义。Wherein, R 1 , R 2 and R 3 are as defined in claim 1. - 根据权利要求1-3任一项所述的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体，其特征在于，The compound according to any one of claims 1-3 or its pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer, characterized in that,R1为氢、氘、卤素或优选氢、 卤素或更优选氢、溴或 R 1 is hydrogen, deuterium, halogen or Preferably hydrogen, Halogen or More preferably hydrogen, bromine orR2为C1-C6烷基，优选C1-C4烷基，更优选甲基；R 2 is C1-C6 alkyl, preferably C1-C4 alkyl, more preferably methyl;R3为C1-C6烷基，优选C1-C4烷基，更优选异丙基。R 3 is C1-C6 alkyl, preferably C1-C4 alkyl, more preferably isopropyl.
- 根据权利要求1-4任一项所述的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体，其特征在于，所述化合物为化合物1、化合物2或化合物3：
The compound according to any one of claims 1 to 4 or its pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer, characterized in that the compound is Compound 1, Compound 2 or Compound 3:
- 一种药物组合物，其包含根据权利要求1-5任一项所述的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体；A pharmaceutical composition comprising a compound according to any one of claims 1-5 or a pharmaceutically acceptable salt, solvate, prodrug, isotopic label or isomer thereof;优选地，所述药物组合物还包含药学上可接受的辅料或载体。Preferably, the pharmaceutical composition further contains pharmaceutically acceptable excipients or carriers.
- 一种药物制剂，其包含根据权利要求1-5任一项所述的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体，或者根据权利要求6所述的药物组合物；A pharmaceutical preparation comprising a compound according to any one of claims 1-5 or a pharmaceutically acceptable salt, solvate, prodrug, isotopic label or isomer thereof, or according to claim 6 pharmaceutical composition;优选地，所述药物制剂为片剂、胶囊剂、注射剂、颗粒剂、粉剂、栓剂、丸剂、凝胶剂、散剂、口服溶液、吸入剂、混悬剂或干悬剂中的任意一种。Preferably, the pharmaceutical preparation is any one of tablets, capsules, injections, granules, powders, suppositories, pills, gels, powders, oral solutions, inhalants, suspensions or dry suspensions.
- 一种药物联合形式，其包含根据权利要求1-5任一项所述的化合物或其药学上可接受的盐、溶剂化物、前药、同位素标记物或异构体，或者根据权利要求6所述的药物组合物，或者根据权利要求7所述的药物制剂，以及预防和/或治疗手足口病的药物中的一种或几种。A pharmaceutical combination form comprising a compound according to any one of claims 1-5 or a pharmaceutically acceptable salt, solvate, prodrug, isotope label or isomer thereof, or a compound according to claim 6 The pharmaceutical composition according to claim 7, or the pharmaceutical preparation according to claim 7, and one or more of the drugs for preventing and/or treating hand, foot and mouth disease.
- 根据权利要求1-5任一项所述的化合物或其药学上可接受的盐、溶剂化物、前药、 同位素标记物或异构体，或者根据权利要求6所述的药物组合物，或者根据权利要求7所述的药物制剂，或者根据权利要求8所述的药物联合形式在制备抗EV71和/或CVA16病毒药物中的应用；The compound according to any one of claims 1-5 or its pharmaceutically acceptable salt, solvate, prodrug, Isotopic markers or isomers, or pharmaceutical compositions according to claim 6, or pharmaceutical preparations according to claim 7, or pharmaceutical combinations according to claim 8 in the preparation of anti-EV71 and/or CVA16 Application in viral drugs;优选地，所述药物为预防和/或治疗至少部分由EV71和/或CVA16病毒引起的疾病和/或病症的药物；Preferably, the medicament is a medicament for preventing and/or treating diseases and/or conditions caused at least in part by EV71 and/or CVA16 viruses;更优选地，所述药物为预防和/或治疗手足口病的药物。 More preferably, the drug is a drug for preventing and/or treating hand, foot and mouth disease.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210933895.9A CN116120282B (en) | 2022-08-04 | 2022-08-04 | Compounds with EV71 and/or CVA16 virus inhibiting activity and application thereof |
| CN202210933895.9 | 2022-08-04 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2024027457A1 true WO2024027457A1 (en) | 2024-02-08 |
Family
ID=86299581
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/CN2023/106356 WO2024027457A1 (en) | 2022-08-04 | 2023-07-07 | Compound having ev71 and/or cva16 virus inhibitory activity and application thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN116120282B (en) |
| WO (1) | WO2024027457A1 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116120282B (en) * | 2022-08-04 | 2024-02-20 | 苏州系统医学研究所 | Compounds with EV71 and/or CVA16 virus inhibiting activity and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090012107A1 (en) * | 2007-01-15 | 2009-01-08 | Mohammad Javad Aman | Antiviral Compounds and Methods of Using Thereof |
| WO2010119050A1 (en) * | 2009-04-14 | 2010-10-21 | Universita' Degli Studi Di Milano | New rac1 inhibitors as potential pharmacological agents for heart failure treatment |
| CN102066339A (en) * | 2008-04-16 | 2011-05-18 | 波托拉医药品公司 | 2, 6-diamino- pyrimidin- 5-yl-carboxamides as SYK or JAK kinases inhibitors |
| US20140323418A1 (en) * | 2011-11-23 | 2014-10-30 | Portola Pharmaceuticals, Inc. | Selective kinase inhibitors |
| CN104744433A (en) * | 2013-12-30 | 2015-07-01 | 中国科学院生物物理研究所 | ALD and application thereof as EV71 virus and CAV16 virus inhibitor |
| CN116120282A (en) * | 2022-08-04 | 2023-05-16 | 苏州系统医学研究所 | Compounds with EV71 and/or CVA16 virus inhibiting activity and application thereof |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2798330A1 (en) * | 2010-05-05 | 2011-11-10 | Infinity Pharmaceuticals, Inc. | Tetrazolones as inhibitors of fatty acid synthase |
| SI2694484T1 (en) * | 2011-04-08 | 2018-10-30 | Janssen Sciences Ireland Uc | Pyrimidine derivatives for the treatment of viral infections |
| US9879003B2 (en) * | 2012-04-11 | 2018-01-30 | Dana-Farber Cancer Institute, Inc. | Host targeted inhibitors of dengue virus and other viruses |
| CA2916623C (en) * | 2013-07-05 | 2021-09-14 | Abivax | Bicyclic compounds useful for treating diseases caused by retroviruses |
-
2022
- 2022-08-04 CN CN202210933895.9A patent/CN116120282B/en active Active
-
2023
- 2023-07-07 WO PCT/CN2023/106356 patent/WO2024027457A1/en unknown
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090012107A1 (en) * | 2007-01-15 | 2009-01-08 | Mohammad Javad Aman | Antiviral Compounds and Methods of Using Thereof |
| CN102066339A (en) * | 2008-04-16 | 2011-05-18 | 波托拉医药品公司 | 2, 6-diamino- pyrimidin- 5-yl-carboxamides as SYK or JAK kinases inhibitors |
| WO2010119050A1 (en) * | 2009-04-14 | 2010-10-21 | Universita' Degli Studi Di Milano | New rac1 inhibitors as potential pharmacological agents for heart failure treatment |
| US20140323418A1 (en) * | 2011-11-23 | 2014-10-30 | Portola Pharmaceuticals, Inc. | Selective kinase inhibitors |
| CN104744433A (en) * | 2013-12-30 | 2015-07-01 | 中国科学院生物物理研究所 | ALD and application thereof as EV71 virus and CAV16 virus inhibitor |
| CN116120282A (en) * | 2022-08-04 | 2023-05-16 | 苏州系统医学研究所 | Compounds with EV71 and/or CVA16 virus inhibiting activity and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| DATABASE REGISTRY 4 February 2014 (2014-02-04), ANONYMOUS: "2,4-Pyrimidinediamine, 5-chloro-N2-methyl-N4-6-quinolinyl- (CA INDEX NAME)", XP093136304, retrieved from STNext Database accession no. 1536927-21-5 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116120282A (en) | 2023-05-16 |
| CN116120282B (en) | 2024-02-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2022143473A1 (en) | Nucleoside compound and use thereof | |
| WO2023173708A1 (en) | Triazine compound or pharmaceutically acceptable salt or isomer thereof, pharmaceutical composition, and use thereof | |
| EP2940031B1 (en) | Nucleoside phosphoramidate compounds for use in the treatment of hcv | |
| US20230105838A1 (en) | Method of Treating COVID-19 | |
| CN109641042B (en) | Heterocyclic derivatives for the treatment of RSV | |
| WO2024027457A1 (en) | Compound having ev71 and/or cva16 virus inhibitory activity and application thereof | |
| CN116751164A (en) | Protease inhibitors, their preparation and use | |
| CN114181258B (en) | Nucleoside compounds for antiviral treatment and application thereof | |
| CN101775003B (en) | Benzothiophene derivative, preparation method and application thereof | |
| CN107459511B (en) | Anti-enterovirus 71(EV71) 4-iminooxazolidine-2-ketone compound and preparation method and application thereof | |
| CN112125920B (en) | Polycyclic benzodifuran compound and application thereof as anti-RSV (respiratory syncytial virus) medicament | |
| CN116693447A (en) | Ketone amide derivative and pharmaceutical application thereof | |
| ES2368089T3 (en) | DERIVATIVES OF 4- (3-BENZOILAMINOFENIL) -6,7-DIMETOXI-2-METHYLAMINOQUINAZOLINE. | |
| CN115160301A (en) | Diphyllin derivative, preparation method and application thereof | |
| WO2021262426A2 (en) | Adamantane amides and thioamides for the treatment of ebolavirus infection | |
| CN114524847B (en) | Anti-hepatitis B virus compound, and preparation method and application thereof | |
| WO2020151296A1 (en) | Dinucleotide precursor for drug and preparation method therefor | |
| CN113149977A (en) | Synthesis and application of respiratory syncytial virus inhibitor | |
| CN116284133B (en) | Novel six-membered heterocyclic derivative, and pharmaceutical composition and application thereof | |
| US11008361B2 (en) | Liver-specific delivery-based anti-hepatitis C prodrug nucleoside cyclo-phosphate compound and uses thereof | |
| CN109134600B (en) | Alkyl and heterocyclic compounds as hepatitis C inhibitors and application thereof in medicines | |
| WO2024078618A1 (en) | Crystal form of cyano-substituted polypeptide compound and preparation method therefor | |
| WO2023185763A1 (en) | Peptidomimetic compound, and preparation method, pharmaceutical composition and use therefor | |
| JP2019530723A (en) | Hepatitis C virus inhibitor and use thereof | |
| JP2010501570A (en) | 4-thio-substituted quinoline and naphthyridine compounds |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 23849166 Country of ref document: EP Kind code of ref document: A1 |