CN115960088A - 一种新型冠状病毒主蛋白酶的抑制剂及其制备方法和用途 - Google Patents
一种新型冠状病毒主蛋白酶的抑制剂及其制备方法和用途 Download PDFInfo
- Publication number
- CN115960088A CN115960088A CN202210944223.8A CN202210944223A CN115960088A CN 115960088 A CN115960088 A CN 115960088A CN 202210944223 A CN202210944223 A CN 202210944223A CN 115960088 A CN115960088 A CN 115960088A
- Authority
- CN
- China
- Prior art keywords
- alkyl
- substituted
- group
- radical
- alkylene
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000002360 preparation method Methods 0.000 title claims abstract description 70
- 241000711573 Coronaviridae Species 0.000 title claims abstract description 30
- 229940125673 3C-like protease inhibitor Drugs 0.000 title description 2
- 150000001875 compounds Chemical class 0.000 claims abstract description 72
- 239000003814 drug Substances 0.000 claims abstract description 32
- 150000003839 salts Chemical class 0.000 claims abstract description 22
- 241001678559 COVID-19 virus Species 0.000 claims abstract description 14
- 230000003287 optical effect Effects 0.000 claims abstract description 14
- 238000006467 substitution reaction Methods 0.000 claims abstract description 9
- 101800000535 3C-like proteinase Proteins 0.000 claims abstract description 8
- 101800002396 3C-like proteinase nsp5 Proteins 0.000 claims abstract description 8
- 230000000155 isotopic effect Effects 0.000 claims abstract description 5
- -1 hydroxy, cyano, amino, carboxy Chemical group 0.000 claims description 73
- 125000000217 alkyl group Chemical group 0.000 claims description 64
- 201000003176 Severe Acute Respiratory Syndrome Diseases 0.000 claims description 36
- 229910052736 halogen Inorganic materials 0.000 claims description 31
- 150000002367 halogens Chemical class 0.000 claims description 31
- 229910052739 hydrogen Inorganic materials 0.000 claims description 28
- 239000001257 hydrogen Substances 0.000 claims description 28
- 125000003545 alkoxy group Chemical group 0.000 claims description 26
- 125000002947 alkylene group Chemical group 0.000 claims description 20
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 20
- 229940079593 drug Drugs 0.000 claims description 19
- 125000005843 halogen group Chemical group 0.000 claims description 18
- 125000003118 aryl group Chemical group 0.000 claims description 15
- 150000002431 hydrogen Chemical class 0.000 claims description 15
- 229910052760 oxygen Inorganic materials 0.000 claims description 15
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 13
- 125000000753 cycloalkyl group Chemical group 0.000 claims description 13
- 125000000623 heterocyclic group Chemical group 0.000 claims description 13
- 229910052717 sulfur Inorganic materials 0.000 claims description 12
- 125000006570 (C5-C6) heteroaryl group Chemical group 0.000 claims description 11
- 125000006297 carbonyl amino group Chemical group [H]N([*:2])C([*:1])=O 0.000 claims description 11
- 208000025721 COVID-19 Diseases 0.000 claims description 10
- 125000005842 heteroatom Chemical group 0.000 claims description 10
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 10
- 125000001424 substituent group Chemical group 0.000 claims description 10
- 108091005804 Peptidases Proteins 0.000 claims description 8
- 208000037847 SARS-CoV-2-infection Diseases 0.000 claims description 8
- 125000006367 bivalent amino carbonyl group Chemical group [H]N([*:1])C([*:2])=O 0.000 claims description 8
- 230000002401 inhibitory effect Effects 0.000 claims description 8
- 102000035195 Peptidases Human genes 0.000 claims description 5
- 201000010099 disease Diseases 0.000 claims description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 5
- YZCKVEUIGOORGS-OUBTZVSYSA-N Deuterium Chemical compound [2H] YZCKVEUIGOORGS-OUBTZVSYSA-N 0.000 claims description 3
- VVBXXVAFSPEIJQ-CVIPOMFBSA-N [(2r)-3-[[(2r)-1-[[(2s,5r,8r,11r,12s,15s,18s,21s)-15-[3-(diaminomethylideneamino)propyl]-21-hydroxy-5-[(4-hydroxyphenyl)methyl]-4,11-dimethyl-2-(2-methylpropyl)-3,6,9,13,16,22-hexaoxo-8-propan-2-yl-10-oxa-1,4,7,14,17-pentazabicyclo[16.3.1]docosan-12-yl]am Chemical compound C([C@@H]1C(=O)N[C@@H](C(=O)O[C@H](C)[C@@H](C(N[C@@H](CCCN=C(N)N)C(=O)N[C@H]2CC[C@H](O)N(C2=O)[C@@H](CC(C)C)C(=O)N1C)=O)NC(=O)[C@H](NC(=O)[C@H](O)COS(O)(=O)=O)CC(C)C)C(C)C)C1=CC=C(O)C=C1 VVBXXVAFSPEIJQ-CVIPOMFBSA-N 0.000 claims description 3
- 239000004480 active ingredient Substances 0.000 claims description 3
- 125000004450 alkenylene group Chemical group 0.000 claims description 3
- 125000004419 alkynylene group Chemical group 0.000 claims description 3
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 claims description 3
- 229910052805 deuterium Inorganic materials 0.000 claims description 3
- 239000008194 pharmaceutical composition Substances 0.000 claims description 3
- 238000011282 treatment Methods 0.000 claims description 3
- 206010035737 Pneumonia viral Diseases 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- 125000001624 naphthyl group Chemical group 0.000 claims description 2
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 claims description 2
- 208000009421 viral pneumonia Diseases 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims 1
- 239000003112 inhibitor Substances 0.000 abstract description 12
- 230000010076 replication Effects 0.000 abstract description 8
- 230000000903 blocking effect Effects 0.000 abstract description 3
- 238000013518 transcription Methods 0.000 abstract description 2
- 230000035897 transcription Effects 0.000 abstract description 2
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 105
- 238000006243 chemical reaction Methods 0.000 description 91
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 78
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 66
- 239000000243 solution Substances 0.000 description 61
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 36
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 34
- 210000004027 cell Anatomy 0.000 description 31
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 30
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 28
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 27
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 27
- 210000004072 lung Anatomy 0.000 description 27
- 239000012074 organic phase Substances 0.000 description 26
- 238000002474 experimental method Methods 0.000 description 24
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 22
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 16
- 239000007787 solid Substances 0.000 description 15
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 14
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 13
- 238000001914 filtration Methods 0.000 description 13
- 238000003756 stirring Methods 0.000 description 13
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 12
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical class [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 12
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 12
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 12
- 238000004440 column chromatography Methods 0.000 description 12
- 239000012043 crude product Substances 0.000 description 12
- 241000700605 Viruses Species 0.000 description 11
- 229910052786 argon Inorganic materials 0.000 description 11
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 10
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 description 10
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 10
- 230000000694 effects Effects 0.000 description 10
- 230000005764 inhibitory process Effects 0.000 description 10
- 210000002540 macrophage Anatomy 0.000 description 10
- GLGNXYJARSMNGJ-VKTIVEEGSA-N (1s,2s,3r,4r)-3-[[5-chloro-2-[(1-ethyl-6-methoxy-2-oxo-4,5-dihydro-3h-1-benzazepin-7-yl)amino]pyrimidin-4-yl]amino]bicyclo[2.2.1]hept-5-ene-2-carboxamide Chemical compound CCN1C(=O)CCCC2=C(OC)C(NC=3N=C(C(=CN=3)Cl)N[C@H]3[C@H]([C@@]4([H])C[C@@]3(C=C4)[H])C(N)=O)=CC=C21 GLGNXYJARSMNGJ-VKTIVEEGSA-N 0.000 description 9
- SZUVGFMDDVSKSI-WIFOCOSTSA-N (1s,2s,3s,5r)-1-(carboxymethyl)-3,5-bis[(4-phenoxyphenyl)methyl-propylcarbamoyl]cyclopentane-1,2-dicarboxylic acid Chemical compound O=C([C@@H]1[C@@H]([C@](CC(O)=O)([C@H](C(=O)N(CCC)CC=2C=CC(OC=3C=CC=CC=3)=CC=2)C1)C(O)=O)C(O)=O)N(CCC)CC(C=C1)=CC=C1OC1=CC=CC=C1 SZUVGFMDDVSKSI-WIFOCOSTSA-N 0.000 description 9
- KAIODGZZEANQLB-UHFFFAOYSA-N 1,2,3,3a,4,5,6,6a-octahydrocyclopenta[c]pyrrole-3-carboxamide Chemical compound C1CCC2C(C(=O)N)NCC21 KAIODGZZEANQLB-UHFFFAOYSA-N 0.000 description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- 241000700159 Rattus Species 0.000 description 9
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 9
- 125000004432 carbon atom Chemical group C* 0.000 description 9
- 229940126543 compound 14 Drugs 0.000 description 9
- 229940125758 compound 15 Drugs 0.000 description 9
- 238000001035 drying Methods 0.000 description 9
- 238000001727 in vivo Methods 0.000 description 9
- OKKJLVBELUTLKV-VMNATFBRSA-N methanol-d1 Chemical compound [2H]OC OKKJLVBELUTLKV-VMNATFBRSA-N 0.000 description 9
- 210000000440 neutrophil Anatomy 0.000 description 9
- 231100000822 oral exposure Toxicity 0.000 description 9
- 239000007800 oxidant agent Substances 0.000 description 9
- 230000002829 reductive effect Effects 0.000 description 9
- 238000011830 transgenic mouse model Methods 0.000 description 9
- 238000005406 washing Methods 0.000 description 9
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 8
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 8
- 239000012279 sodium borohydride Substances 0.000 description 8
- 229910000033 sodium borohydride Inorganic materials 0.000 description 8
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 7
- 239000012964 benzotriazole Substances 0.000 description 7
- 230000015572 biosynthetic process Effects 0.000 description 7
- 238000007912 intraperitoneal administration Methods 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- 229910052757 nitrogen Inorganic materials 0.000 description 7
- 238000003786 synthesis reaction Methods 0.000 description 7
- WORJRXHJTUTINR-UHFFFAOYSA-N 1,4-dioxane;hydron;chloride Chemical compound Cl.C1COCCO1 WORJRXHJTUTINR-UHFFFAOYSA-N 0.000 description 6
- 102000019034 Chemokines Human genes 0.000 description 6
- 108010012236 Chemokines Proteins 0.000 description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 6
- 241000699660 Mus musculus Species 0.000 description 6
- 239000008346 aqueous phase Substances 0.000 description 6
- 238000003556 assay Methods 0.000 description 6
- 125000004429 atom Chemical group 0.000 description 6
- 229940126214 compound 3 Drugs 0.000 description 6
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 6
- 238000011156 evaluation Methods 0.000 description 6
- 238000000605 extraction Methods 0.000 description 6
- 239000003208 petroleum Substances 0.000 description 6
- 125000003367 polycyclic group Chemical group 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 125000002914 sec-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 6
- 125000006633 tert-butoxycarbonylamino group Chemical group 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 5
- 102000004127 Cytokines Human genes 0.000 description 5
- 108090000695 Cytokines Proteins 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 239000007864 aqueous solution Substances 0.000 description 5
- 230000014509 gene expression Effects 0.000 description 5
- 208000015181 infectious disease Diseases 0.000 description 5
- 230000002757 inflammatory effect Effects 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 230000001590 oxidative effect Effects 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 230000029812 viral genome replication Effects 0.000 description 5
- 230000003612 virological effect Effects 0.000 description 5
- UDQTXCHQKHIQMH-KYGLGHNPSA-N (3ar,5s,6s,7r,7ar)-5-(difluoromethyl)-2-(ethylamino)-5,6,7,7a-tetrahydro-3ah-pyrano[3,2-d][1,3]thiazole-6,7-diol Chemical compound S1C(NCC)=N[C@H]2[C@@H]1O[C@H](C(F)F)[C@@H](O)[C@@H]2O UDQTXCHQKHIQMH-KYGLGHNPSA-N 0.000 description 4
- MPDDTAJMJCESGV-CTUHWIOQSA-M (3r,5r)-7-[2-(4-fluorophenyl)-5-[methyl-[(1r)-1-phenylethyl]carbamoyl]-4-propan-2-ylpyrazol-3-yl]-3,5-dihydroxyheptanoate Chemical compound C1([C@@H](C)N(C)C(=O)C2=NN(C(CC[C@@H](O)C[C@@H](O)CC([O-])=O)=C2C(C)C)C=2C=CC(F)=CC=2)=CC=CC=C1 MPDDTAJMJCESGV-CTUHWIOQSA-M 0.000 description 4
- 241000699670 Mus sp. Species 0.000 description 4
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 4
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 4
- 210000002821 alveolar epithelial cell Anatomy 0.000 description 4
- 230000030833 cell death Effects 0.000 description 4
- 229940125904 compound 1 Drugs 0.000 description 4
- 229940125936 compound 42 Drugs 0.000 description 4
- 238000001704 evaporation Methods 0.000 description 4
- 125000001072 heteroaryl group Chemical group 0.000 description 4
- 210000004969 inflammatory cell Anatomy 0.000 description 4
- ZCSHNCUQKCANBX-UHFFFAOYSA-N lithium diisopropylamide Chemical compound [Li+].CC(C)[N-]C(C)C ZCSHNCUQKCANBX-UHFFFAOYSA-N 0.000 description 4
- RUZLIIJDZBWWSA-INIZCTEOSA-N methyl 2-[[(1s)-1-(7-methyl-2-morpholin-4-yl-4-oxopyrido[1,2-a]pyrimidin-9-yl)ethyl]amino]benzoate Chemical group COC(=O)C1=CC=CC=C1N[C@@H](C)C1=CC(C)=CN2C(=O)C=C(N3CCOCC3)N=C12 RUZLIIJDZBWWSA-INIZCTEOSA-N 0.000 description 4
- 238000012544 monitoring process Methods 0.000 description 4
- 239000012071 phase Substances 0.000 description 4
- 238000010791 quenching Methods 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 238000000967 suction filtration Methods 0.000 description 4
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 4
- WVDDGKGOMKODPV-UHFFFAOYSA-N Benzyl alcohol Chemical compound OCC1=CC=CC=C1 WVDDGKGOMKODPV-UHFFFAOYSA-N 0.000 description 3
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- 239000007821 HATU Substances 0.000 description 3
- 101000656751 Haloarcula marismortui (strain ATCC 43049 / DSM 3752 / JCM 8966 / VKM B-1809) 30S ribosomal protein S24e Proteins 0.000 description 3
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 239000004365 Protease Substances 0.000 description 3
- 238000011529 RT qPCR Methods 0.000 description 3
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 3
- 230000000840 anti-viral effect Effects 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 239000000460 chlorine Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 239000012065 filter cake Substances 0.000 description 3
- 238000007489 histopathology method Methods 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- UEXQBEVWFZKHNB-UHFFFAOYSA-N intermediate 29 Natural products C1=CC(N)=CC=C1NC1=NC=CC=N1 UEXQBEVWFZKHNB-UHFFFAOYSA-N 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Substances N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 3
- 239000012044 organic layer Substances 0.000 description 3
- 230000001575 pathological effect Effects 0.000 description 3
- 238000004237 preparative chromatography Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 230000000171 quenching effect Effects 0.000 description 3
- 229910052938 sodium sulfate Inorganic materials 0.000 description 3
- 235000011152 sodium sulphate Nutrition 0.000 description 3
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 3
- 235000019345 sodium thiosulphate Nutrition 0.000 description 3
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 125000006318 tert-butyl amino group Chemical group [H]N(*)C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H] 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- DPEMLPYPRWNMFD-QEORTHHSSA-N (1R,2S,5S)-1,6,6-trimethyl-3-[2-[4-(trifluoromethoxy)phenoxy]acetyl]-3-azabicyclo[3.1.0]hexane-2-carboxylic acid Chemical compound CC(C)([C@@H]1C2)[C@]1(C)[C@@H](C(O)=O)N2C(COC(C=C1)=CC=C1OC(F)(F)F)=O DPEMLPYPRWNMFD-QEORTHHSSA-N 0.000 description 2
- SUCZQTHNYBDZQG-GEMLJDPKSA-N (3s)-3-[(2s)-2-amino-4-chloro-3-oxobutyl]pyrrolidin-2-one;hydrochloride Chemical compound Cl.ClCC(=O)[C@@H](N)C[C@@H]1CCNC1=O SUCZQTHNYBDZQG-GEMLJDPKSA-N 0.000 description 2
- KWRNCAUXSJOYQO-UHFFFAOYSA-N 1,2,3,3a,4,5,6,6a-octahydrocyclopenta[c]pyrrol-2-ium-3-carboxylate Chemical compound C1CCC2C(C(=O)O)NCC21 KWRNCAUXSJOYQO-UHFFFAOYSA-N 0.000 description 2
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 2
- YSUIQYOGTINQIN-UZFYAQMZSA-N 2-amino-9-[(1S,6R,8R,9S,10R,15R,17R,18R)-8-(6-aminopurin-9-yl)-9,18-difluoro-3,12-dihydroxy-3,12-bis(sulfanylidene)-2,4,7,11,13,16-hexaoxa-3lambda5,12lambda5-diphosphatricyclo[13.2.1.06,10]octadecan-17-yl]-1H-purin-6-one Chemical compound NC1=NC2=C(N=CN2[C@@H]2O[C@@H]3COP(S)(=O)O[C@@H]4[C@@H](COP(S)(=O)O[C@@H]2[C@@H]3F)O[C@H]([C@H]4F)N2C=NC3=C2N=CN=C3N)C(=O)N1 YSUIQYOGTINQIN-UZFYAQMZSA-N 0.000 description 2
- PVAZVBVIWJGNJA-UHFFFAOYSA-N 2-fluoro-3-oxo-3-phenylmethoxypropanoic acid Chemical compound OC(=O)C(F)C(=O)OCC1=CC=CC=C1 PVAZVBVIWJGNJA-UHFFFAOYSA-N 0.000 description 2
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 2
- 210000003771 C cell Anatomy 0.000 description 2
- OABHWGXRPRKKOQ-QEJZJMRPSA-N CC(C)([C@H]1C2)[C@@H]1[C@@H](C(NC)=O)N2C(COC(C=C1)=CC=C1OC(F)(F)F)=O Chemical compound CC(C)([C@H]1C2)[C@@H]1[C@@H](C(NC)=O)N2C(COC(C=C1)=CC=C1OC(F)(F)F)=O OABHWGXRPRKKOQ-QEJZJMRPSA-N 0.000 description 2
- KXDHJXZQYSOELW-UHFFFAOYSA-M Carbamate Chemical compound NC([O-])=O KXDHJXZQYSOELW-UHFFFAOYSA-M 0.000 description 2
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 2
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- HCUARRIEZVDMPT-UHFFFAOYSA-N Indole-2-carboxylic acid Chemical compound C1=CC=C2NC(C(=O)O)=CC2=C1 HCUARRIEZVDMPT-UHFFFAOYSA-N 0.000 description 2
- 102100026720 Interferon beta Human genes 0.000 description 2
- 108090000467 Interferon-beta Proteins 0.000 description 2
- 108700026244 Open Reading Frames Proteins 0.000 description 2
- 101800004803 Papain-like protease Proteins 0.000 description 2
- 229930040373 Paraformaldehyde Natural products 0.000 description 2
- 108010076039 Polyproteins Proteins 0.000 description 2
- 101710153041 Replicase polyprotein 1a Proteins 0.000 description 2
- 101710151619 Replicase polyprotein 1ab Proteins 0.000 description 2
- 108010087230 Sincalide Proteins 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical class [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- 101710172711 Structural protein Proteins 0.000 description 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical group [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 2
- 108010065667 Viral Matrix Proteins Proteins 0.000 description 2
- 108020000999 Viral RNA Proteins 0.000 description 2
- OZQXEOSNFMMMRD-UHFFFAOYSA-M [Cl-].CC(C)[Mg+].C1CCOC1 Chemical compound [Cl-].CC(C)[Mg+].C1CCOC1 OZQXEOSNFMMMRD-UHFFFAOYSA-M 0.000 description 2
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 125000003342 alkenyl group Chemical group 0.000 description 2
- 125000000304 alkynyl group Chemical group 0.000 description 2
- 235000019270 ammonium chloride Nutrition 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical group [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- PFKFTWBEEFSNDU-UHFFFAOYSA-N carbonyldiimidazole Chemical compound C1=CN=CN1C(=O)N1C=CN=C1 PFKFTWBEEFSNDU-UHFFFAOYSA-N 0.000 description 2
- 239000001768 carboxy methyl cellulose Substances 0.000 description 2
- 238000010609 cell counting kit-8 assay Methods 0.000 description 2
- 229910052801 chlorine Inorganic materials 0.000 description 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical class OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 2
- GFHNAMRJFCEERV-UHFFFAOYSA-L cobalt chloride hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].[Cl-].[Co+2] GFHNAMRJFCEERV-UHFFFAOYSA-L 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- FWNITJYYNOTRCY-UHFFFAOYSA-N dibenzyl 2-fluoropropanedioate Chemical compound C=1C=CC=CC=1COC(=O)C(F)C(=O)OCC1=CC=CC=C1 FWNITJYYNOTRCY-UHFFFAOYSA-N 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- QNSPKWUAZQIIGZ-QMMMGPOBSA-N dimethyl (2s)-2-[(2-methylpropan-2-yl)oxycarbonylamino]pentanedioate Chemical compound COC(=O)CC[C@@H](C(=O)OC)NC(=O)OC(C)(C)C QNSPKWUAZQIIGZ-QMMMGPOBSA-N 0.000 description 2
- PQVSTLUFSYVLTO-UHFFFAOYSA-N ethyl n-ethoxycarbonylcarbamate Chemical compound CCOC(=O)NC(=O)OCC PQVSTLUFSYVLTO-UHFFFAOYSA-N 0.000 description 2
- 239000011737 fluorine Substances 0.000 description 2
- 229910052731 fluorine Inorganic materials 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- 230000008595 infiltration Effects 0.000 description 2
- 238000001764 infiltration Methods 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- GLXDVVHUTZTUQK-UHFFFAOYSA-M lithium hydroxide monohydrate Substances [Li+].O.[OH-] GLXDVVHUTZTUQK-UHFFFAOYSA-M 0.000 description 2
- 229940040692 lithium hydroxide monohydrate Drugs 0.000 description 2
- BYRPTKZOXNFFDB-UHFFFAOYSA-N lithium;bis(trimethylsilyl)azanide;oxolane Chemical compound [Li+].C1CCOC1.C[Si](C)(C)[N-][Si](C)(C)C BYRPTKZOXNFFDB-UHFFFAOYSA-N 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- UKVIEHSSVKSQBA-UHFFFAOYSA-N methane;palladium Chemical compound C.[Pd] UKVIEHSSVKSQBA-UHFFFAOYSA-N 0.000 description 2
- 238000000386 microscopy Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 125000002950 monocyclic group Chemical group 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 239000001301 oxygen Chemical group 0.000 description 2
- 239000012188 paraffin wax Substances 0.000 description 2
- 229920002866 paraformaldehyde Polymers 0.000 description 2
- 229960005190 phenylalanine Drugs 0.000 description 2
- 239000000902 placebo Substances 0.000 description 2
- 229940068196 placebo Drugs 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- REQCZEXYDRLIBE-UHFFFAOYSA-N procainamide Chemical compound CCN(CC)CCNC(=O)C1=CC=C(N)C=C1 REQCZEXYDRLIBE-UHFFFAOYSA-N 0.000 description 2
- 229960002429 proline Drugs 0.000 description 2
- DYWOPZYICSJYMT-UHFFFAOYSA-N propanoic acid;2,2,2-trifluoroacetic acid Chemical compound CCC(O)=O.OC(=O)C(F)(F)F DYWOPZYICSJYMT-UHFFFAOYSA-N 0.000 description 2
- 230000002685 pulmonary effect Effects 0.000 description 2
- FDDQRDMHICUGQC-UHFFFAOYSA-M pyrrole-1-carboxylate Chemical compound [O-]C(=O)N1C=CC=C1 FDDQRDMHICUGQC-UHFFFAOYSA-M 0.000 description 2
- FDDQRDMHICUGQC-UHFFFAOYSA-N pyrrole-1-carboxylic acid Chemical compound OC(=O)N1C=CC=C1 FDDQRDMHICUGQC-UHFFFAOYSA-N 0.000 description 2
- 238000003762 quantitative reverse transcription PCR Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 125000006413 ring segment Chemical group 0.000 description 2
- 229920006395 saturated elastomer Polymers 0.000 description 2
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 2
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000012453 sprague-dawley rat model Methods 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 2
- 239000011593 sulfur Chemical group 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 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
- 230000009385 viral infection Effects 0.000 description 2
- GUYTVDGFJRFUGN-KHYOSLBOSA-N (1R,2S,5S)-1,6,6-trimethyl-3-(quinoline-2-carbonyl)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid Chemical compound CC(C)([C@@H]1C2)[C@]1(C)[C@@H](C(O)=O)N2C(C1=NC2=CC=CC=C2C=C1)=O GUYTVDGFJRFUGN-KHYOSLBOSA-N 0.000 description 1
- OPWWOGRDBZRHNN-CQDKDKBSSA-N (1R,2S,5S)-6,6-dimethyl-3-(quinoline-2-carbonyl)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid Chemical compound CC(C)([C@H]1C2)[C@@H]1[C@@H](C(O)=O)N2C(C1=NC2=CC=CC=C2C=C1)=O OPWWOGRDBZRHNN-CQDKDKBSSA-N 0.000 description 1
- USAVFMVBIUUZTF-UBHSHLNASA-N (1R,2S,5S)-6,6-dimethyl-3-[2-[4-(trifluoromethoxy)phenoxy]acetyl]-3-azabicyclo[3.1.0]hexane-2-carboxylic acid Chemical compound CC(C)([C@H]1C2)[C@@H]1[C@@H](C(O)=O)N2C(COC(C=C1)=CC=C1OC(F)(F)F)=O USAVFMVBIUUZTF-UBHSHLNASA-N 0.000 description 1
- ASGMFNBUXDJWJJ-JLCFBVMHSA-N (1R,3R)-3-[[3-bromo-1-[4-(5-methyl-1,3,4-thiadiazol-2-yl)phenyl]pyrazolo[3,4-d]pyrimidin-6-yl]amino]-N,1-dimethylcyclopentane-1-carboxamide Chemical compound BrC1=NN(C2=NC(=NC=C21)N[C@H]1C[C@@](CC1)(C(=O)NC)C)C1=CC=C(C=C1)C=1SC(=NN=1)C ASGMFNBUXDJWJJ-JLCFBVMHSA-N 0.000 description 1
- IWKFBGQBNHOVTD-AVMPFWSBSA-N (1R,5S)-6,6-dimethyl-N-[(2S)-1-oxo-3-phenylpropan-2-yl]-3-(quinoline-2-carbonyl)-3-azabicyclo[3.1.0]hexane-2-carboxamide Chemical compound CC(C)([C@H]1C2)[C@@H]1C(C(N[C@@H](CC1=CC=CC=C1)C=O)=O)N2C(C1=NC2=CC=CC=C2C=C1)=O IWKFBGQBNHOVTD-AVMPFWSBSA-N 0.000 description 1
- RWCCIYJJIFJDQO-YUMQZZPRSA-N (2S)-2-[(2-methylpropan-2-yl)oxycarbonylamino]-3-[(3S)-2-oxopyrrolidin-3-yl]propanoic acid Chemical compound CC(C)(C)OC(=O)N[C@H](C(O)=O)C[C@@H]1CCNC1=O RWCCIYJJIFJDQO-YUMQZZPRSA-N 0.000 description 1
- IUSARDYWEPUTPN-OZBXUNDUSA-N (2r)-n-[(2s,3r)-4-[[(4s)-6-(2,2-dimethylpropyl)spiro[3,4-dihydropyrano[2,3-b]pyridine-2,1'-cyclobutane]-4-yl]amino]-3-hydroxy-1-[3-(1,3-thiazol-2-yl)phenyl]butan-2-yl]-2-methoxypropanamide Chemical compound C([C@H](NC(=O)[C@@H](C)OC)[C@H](O)CN[C@@H]1C2=CC(CC(C)(C)C)=CN=C2OC2(CCC2)C1)C(C=1)=CC=CC=1C1=NC=CS1 IUSARDYWEPUTPN-OZBXUNDUSA-N 0.000 description 1
- WFPSMPYVXFVVFA-LURJTMIESA-N (2s)-4-methoxy-2-[(2-methylpropan-2-yl)oxycarbonylamino]-4-oxobutanoic acid Chemical compound COC(=O)C[C@@H](C(O)=O)NC(=O)OC(C)(C)C WFPSMPYVXFVVFA-LURJTMIESA-N 0.000 description 1
- HUWSZNZAROKDRZ-RRLWZMAJSA-N (3r,4r)-3-azaniumyl-5-[[(2s,3r)-1-[(2s)-2,3-dicarboxypyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl]amino]-5-oxo-4-sulfanylpentane-1-sulfonate Chemical compound OS(=O)(=O)CC[C@@H](N)[C@@H](S)C(=O)N[C@@H]([C@H](C)CC)C(=O)N1CCC(C(O)=O)[C@H]1C(O)=O HUWSZNZAROKDRZ-RRLWZMAJSA-N 0.000 description 1
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 1
- WZZBNLYBHUDSHF-DHLKQENFSA-N 1-[(3s,4s)-4-[8-(2-chloro-4-pyrimidin-2-yloxyphenyl)-7-fluoro-2-methylimidazo[4,5-c]quinolin-1-yl]-3-fluoropiperidin-1-yl]-2-hydroxyethanone Chemical compound CC1=NC2=CN=C3C=C(F)C(C=4C(=CC(OC=5N=CC=CN=5)=CC=4)Cl)=CC3=C2N1[C@H]1CCN(C(=O)CO)C[C@@H]1F WZZBNLYBHUDSHF-DHLKQENFSA-N 0.000 description 1
- MOSJNBGPCBBNCG-UHFFFAOYSA-N 2,2,3,3-tetramethylbutanenitrile Chemical compound CC(C)(C)C(C)(C)C#N MOSJNBGPCBBNCG-UHFFFAOYSA-N 0.000 description 1
- 239000005631 2,4-Dichlorophenoxyacetic acid Substances 0.000 description 1
- HXKWSTRRCHTUEC-UHFFFAOYSA-N 2,4-Dichlorophenoxyaceticacid Chemical compound OC(=O)C(Cl)OC1=CC=C(Cl)C=C1 HXKWSTRRCHTUEC-UHFFFAOYSA-N 0.000 description 1
- FQMZXMVHHKXGTM-UHFFFAOYSA-N 2-(1-adamantyl)-n-[2-[2-(2-hydroxyethylamino)ethylamino]quinolin-5-yl]acetamide Chemical compound C1C(C2)CC(C3)CC2CC13CC(=O)NC1=CC=CC2=NC(NCCNCCO)=CC=C21 FQMZXMVHHKXGTM-UHFFFAOYSA-N 0.000 description 1
- QHSBEEUEIRDHCD-UHFFFAOYSA-N 2-[4-(trifluoromethoxy)phenoxy]acetic acid Chemical compound OC(=O)COC1=CC=C(OC(F)(F)F)C=C1 QHSBEEUEIRDHCD-UHFFFAOYSA-N 0.000 description 1
- PYRKKGOKRMZEIT-UHFFFAOYSA-N 2-[6-(2-cyclopropylethoxy)-9-(2-hydroxy-2-methylpropyl)-1h-phenanthro[9,10-d]imidazol-2-yl]-5-fluorobenzene-1,3-dicarbonitrile Chemical compound C1=C2C3=CC(CC(C)(O)C)=CC=C3C=3NC(C=4C(=CC(F)=CC=4C#N)C#N)=NC=3C2=CC=C1OCCC1CC1 PYRKKGOKRMZEIT-UHFFFAOYSA-N 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- REXUYBKPWIPONM-UHFFFAOYSA-N 2-bromoacetonitrile Chemical compound BrCC#N REXUYBKPWIPONM-UHFFFAOYSA-N 0.000 description 1
- WLAMNBDJUVNPJU-UHFFFAOYSA-N 2-methylbutyric acid Chemical compound CCC(C)C(O)=O WLAMNBDJUVNPJU-UHFFFAOYSA-N 0.000 description 1
- CVKMFSAVYPAZTQ-UHFFFAOYSA-N 2-methylhexanoic acid Chemical compound CCCCC(C)C(O)=O CVKMFSAVYPAZTQ-UHFFFAOYSA-N 0.000 description 1
- 101800000504 3C-like protease Proteins 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
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 102100030988 Angiotensin-converting enzyme Human genes 0.000 description 1
- 102100035765 Angiotensin-converting enzyme 2 Human genes 0.000 description 1
- 108090000975 Angiotensin-converting enzyme 2 Proteins 0.000 description 1
- 101000666833 Autographa californica nuclear polyhedrosis virus Uncharacterized 20.8 kDa protein in FGF-VUBI intergenic region Proteins 0.000 description 1
- 101000977027 Azospirillum brasilense Uncharacterized protein in nodG 5'region Proteins 0.000 description 1
- 101000962005 Bacillus thuringiensis Uncharacterized 23.6 kDa protein Proteins 0.000 description 1
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 1
- ISMDILRWKSYCOD-GNKBHMEESA-N C(C1=CC=CC=C1)[C@@H]1NC(OCCCCCCCCCCCNC([C@@H](NC(C[C@@H]1O)=O)C(C)C)=O)=O Chemical compound C(C1=CC=CC=C1)[C@@H]1NC(OCCCCCCCCCCCNC([C@@H](NC(C[C@@H]1O)=O)C(C)C)=O)=O ISMDILRWKSYCOD-GNKBHMEESA-N 0.000 description 1
- WKOVWKJWJUMKCO-IUCAKERBSA-N CC(C)(C)OC(N[C@@H](C[C@H](CCN1)C1=C=O)C(O)=O)=O Chemical compound CC(C)(C)OC(N[C@@H](C[C@H](CCN1)C1=C=O)C(O)=O)=O WKOVWKJWJUMKCO-IUCAKERBSA-N 0.000 description 1
- JBBLNCKOVRJHOL-BYPYZUCNSA-N COC(C[C@@H](C(CF)=O)N)=O Chemical compound COC(C[C@@H](C(CF)=O)N)=O JBBLNCKOVRJHOL-BYPYZUCNSA-N 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 229940126639 Compound 33 Drugs 0.000 description 1
- 229940127007 Compound 39 Drugs 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
- ZAFNJMIOTHYJRJ-UHFFFAOYSA-N Diisopropyl ether Chemical compound CC(C)OC(C)C ZAFNJMIOTHYJRJ-UHFFFAOYSA-N 0.000 description 1
- 101000785191 Drosophila melanogaster Uncharacterized 50 kDa protein in type I retrotransposable element R1DM Proteins 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 101000747704 Enterobacteria phage N4 Uncharacterized protein Gp1 Proteins 0.000 description 1
- 101000861206 Enterococcus faecalis (strain ATCC 700802 / V583) Uncharacterized protein EF_A0048 Proteins 0.000 description 1
- 101000769180 Escherichia coli Uncharacterized 11.1 kDa protein Proteins 0.000 description 1
- KRHYYFGTRYWZRS-UHFFFAOYSA-N Fluorane Chemical compound F KRHYYFGTRYWZRS-UHFFFAOYSA-N 0.000 description 1
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 1
- 101800003471 Helicase Proteins 0.000 description 1
- 101800002870 Helicase nsp13 Proteins 0.000 description 1
- 101000833492 Homo sapiens Jouberin Proteins 0.000 description 1
- 101000651236 Homo sapiens NCK-interacting protein with SH3 domain Proteins 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-N Hydrogen bromide Chemical compound Br CPELXLSAUQHCOX-UHFFFAOYSA-N 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 102100024407 Jouberin Human genes 0.000 description 1
- 102100034866 Kallikrein-6 Human genes 0.000 description 1
- 101000976301 Leptospira interrogans Uncharacterized 35 kDa protein in sph 3'region Proteins 0.000 description 1
- JYOAXOMPIXKMKK-YUMQZZPRSA-N Leu-Gln Chemical compound CC(C)C[C@H]([NH3+])C(=O)N[C@H](C([O-])=O)CCC(N)=O JYOAXOMPIXKMKK-YUMQZZPRSA-N 0.000 description 1
- 208000004852 Lung Injury Diseases 0.000 description 1
- UBOFRNNKOVPWBW-WDSKDSINSA-N N[C@@H](C[C@H](CCN1)C1=O)C(CF)=O Chemical compound N[C@@H](C[C@H](CCN1)C1=O)C(CF)=O UBOFRNNKOVPWBW-WDSKDSINSA-N 0.000 description 1
- 101000658690 Neisseria meningitidis serogroup B Transposase for insertion sequence element IS1106 Proteins 0.000 description 1
- 101800000934 Non-structural protein 13 Proteins 0.000 description 1
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 1
- 101150001779 ORF1a gene Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 101800001016 Picornain 3C-like protease Proteins 0.000 description 1
- 101800000596 Probable picornain 3C-like protease Proteins 0.000 description 1
- YZCKVEUIGOORGS-IGMARMGPSA-N Protium Chemical compound [1H] YZCKVEUIGOORGS-IGMARMGPSA-N 0.000 description 1
- 241000720974 Protium Species 0.000 description 1
- 101000933967 Pseudomonas phage KPP25 Major capsid protein Proteins 0.000 description 1
- 101000748660 Pseudomonas savastanoi Uncharacterized 21 kDa protein in iaaL 5'region Proteins 0.000 description 1
- 101710118046 RNA-directed RNA polymerase Proteins 0.000 description 1
- 101000584469 Rice tungro bacilliform virus (isolate Philippines) Protein P1 Proteins 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 101000818096 Spirochaeta aurantia Uncharacterized 15.5 kDa protein in trpE 3'region Proteins 0.000 description 1
- 101000766081 Streptomyces ambofaciens Uncharacterized HTH-type transcriptional regulator in unstable DNA locus Proteins 0.000 description 1
- 108091027544 Subgenomic mRNA Proteins 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 101000804403 Synechococcus elongatus (strain PCC 7942 / FACHB-805) Uncharacterized HIT-like protein Synpcc7942_1390 Proteins 0.000 description 1
- 101000750910 Synechococcus elongatus (strain PCC 7942 / FACHB-805) Uncharacterized HTH-type transcriptional regulator Synpcc7942_2319 Proteins 0.000 description 1
- 101000644897 Synechococcus sp. (strain ATCC 27264 / PCC 7002 / PR-6) Uncharacterized protein SYNPCC7002_B0001 Proteins 0.000 description 1
- 206010069363 Traumatic lung injury Diseases 0.000 description 1
- YZCKVEUIGOORGS-NJFSPNSNSA-N Tritium Chemical compound [3H] YZCKVEUIGOORGS-NJFSPNSNSA-N 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 101000916336 Xenopus laevis Transposon TX1 uncharacterized 82 kDa protein Proteins 0.000 description 1
- 101001000760 Zea mays Putative Pol polyprotein from transposon element Bs1 Proteins 0.000 description 1
- 101000678262 Zymomonas mobilis subsp. mobilis (strain ATCC 10988 / DSM 424 / LMG 404 / NCIMB 8938 / NRRL B-806 / ZM1) 65 kDa protein Proteins 0.000 description 1
- PSLUFJFHTBIXMW-WYEYVKMPSA-N [(3r,4ar,5s,6s,6as,10s,10ar,10bs)-3-ethenyl-10,10b-dihydroxy-3,4a,7,7,10a-pentamethyl-1-oxo-6-(2-pyridin-2-ylethylcarbamoyloxy)-5,6,6a,8,9,10-hexahydro-2h-benzo[f]chromen-5-yl] acetate Chemical compound O([C@@H]1[C@@H]([C@]2(O[C@](C)(CC(=O)[C@]2(O)[C@@]2(C)[C@@H](O)CCC(C)(C)[C@@H]21)C=C)C)OC(=O)C)C(=O)NCCC1=CC=CC=N1 PSLUFJFHTBIXMW-WYEYVKMPSA-N 0.000 description 1
- 125000005210 alkyl ammonium group Chemical group 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 239000012736 aqueous medium Substances 0.000 description 1
- 239000012131 assay buffer Substances 0.000 description 1
- 229940077388 benzenesulfonate Drugs 0.000 description 1
- SRSXLGNVWSONIS-UHFFFAOYSA-M benzenesulfonate Chemical compound [O-]S(=O)(=O)C1=CC=CC=C1 SRSXLGNVWSONIS-UHFFFAOYSA-M 0.000 description 1
- 235000019445 benzyl alcohol Nutrition 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 238000010241 blood sampling Methods 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 1
- 229910052794 bromium Inorganic materials 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- NEHMKBQYUWJMIP-NJFSPNSNSA-N chloro(114C)methane Chemical compound [14CH3]Cl NEHMKBQYUWJMIP-NJFSPNSNSA-N 0.000 description 1
- PJGJQVRXEUVAFT-UHFFFAOYSA-N chloroiodomethane Chemical compound ClCI PJGJQVRXEUVAFT-UHFFFAOYSA-N 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 239000012230 colorless oil Substances 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 229940125807 compound 37 Drugs 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- QVTOOWHELURIDU-UWVGGRQHSA-N dimethyl (2r,4s)-2-(cyanomethyl)-4-[(2-methylpropan-2-yl)oxycarbonylamino]pentanedioate Chemical compound COC(=O)[C@@H](CC#N)C[C@@H](C(=O)OC)NC(=O)OC(C)(C)C QVTOOWHELURIDU-UWVGGRQHSA-N 0.000 description 1
- ZVXHZSXYHFBIEW-UHFFFAOYSA-N dimethyl 2-fluoropropanedioate Chemical compound COC(=O)C(F)C(=O)OC ZVXHZSXYHFBIEW-UHFFFAOYSA-N 0.000 description 1
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 239000013583 drug formulation Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- VFRSADQPWYCXDG-LEUCUCNGSA-N ethyl (2s,5s)-5-methylpyrrolidine-2-carboxylate;2,2,2-trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F.CCOC(=O)[C@@H]1CC[C@H](C)N1 VFRSADQPWYCXDG-LEUCUCNGSA-N 0.000 description 1
- 125000004494 ethyl ester group Chemical group 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- ZHNUHDYFZUAESO-UHFFFAOYSA-N formamide Substances NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000037433 frameshift Effects 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 125000002541 furyl group Chemical group 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- ZFGMDIBRIDKWMY-PASTXAENSA-N heparin Chemical compound CC(O)=N[C@@H]1[C@@H](O)[C@H](O)[C@@H](COS(O)(=O)=O)O[C@@H]1O[C@@H]1[C@@H](C(O)=O)O[C@@H](O[C@H]2[C@@H]([C@@H](OS(O)(=O)=O)[C@@H](O[C@@H]3[C@@H](OC(O)[C@H](OS(O)(=O)=O)[C@H]3O)C(O)=O)O[C@@H]2O)CS(O)(=O)=O)[C@H](O)[C@H]1O ZFGMDIBRIDKWMY-PASTXAENSA-N 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 229960001008 heparin sodium Drugs 0.000 description 1
- 150000002430 hydrocarbons Chemical group 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 229960001388 interferon-beta Drugs 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 210000004731 jugular vein Anatomy 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 231100000515 lung injury Toxicity 0.000 description 1
- GHFGJTVYMNRGBY-UHFFFAOYSA-N m-tyramine Chemical compound NCCC1=CC=CC(O)=C1 GHFGJTVYMNRGBY-UHFFFAOYSA-N 0.000 description 1
- 229940049920 malate Drugs 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
- BJEPYKJPYRNKOW-UHFFFAOYSA-N malic acid Chemical compound OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- FKVUDBWXNAFSPB-MKXDVQRUSA-N methyl (1r,2s,5s)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylate;hydrochloride Chemical compound Cl.COC(=O)[C@H]1NC[C@@H]2C(C)(C)[C@H]12 FKVUDBWXNAFSPB-MKXDVQRUSA-N 0.000 description 1
- HQEIPVHJHZTMDP-JEDNCBNOSA-N methyl (2s)-pyrrolidine-2-carboxylate;hydrochloride Chemical compound Cl.COC(=O)[C@@H]1CCCN1 HQEIPVHJHZTMDP-JEDNCBNOSA-N 0.000 description 1
- BAAQIHRVIIQRMV-UHFFFAOYSA-N methyl 5-fluoro-4-oxopentanoate Chemical compound COC(=O)CCC(=O)CF BAAQIHRVIIQRMV-UHFFFAOYSA-N 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- DUWWHGPELOTTOE-UHFFFAOYSA-N n-(5-chloro-2,4-dimethoxyphenyl)-3-oxobutanamide Chemical compound COC1=CC(OC)=C(NC(=O)CC(C)=O)C=C1Cl DUWWHGPELOTTOE-UHFFFAOYSA-N 0.000 description 1
- UPSFMJHZUCSEHU-JYGUBCOQSA-N n-[(2s,3r,4r,5s,6r)-2-[(2r,3s,4r,5r,6s)-5-acetamido-4-hydroxy-2-(hydroxymethyl)-6-(4-methyl-2-oxochromen-7-yl)oxyoxan-3-yl]oxy-4,5-dihydroxy-6-(hydroxymethyl)oxan-3-yl]acetamide Chemical compound CC(=O)N[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@H]1[C@H](O)[C@@H](NC(C)=O)[C@H](OC=2C=C3OC(=O)C=C(C)C3=CC=2)O[C@@H]1CO UPSFMJHZUCSEHU-JYGUBCOQSA-N 0.000 description 1
- 230000018791 negative regulation of catalytic activity Effects 0.000 description 1
- 230000003448 neutrophilic effect Effects 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- WVDDGKGOMKODPV-ZQBYOMGUSA-N phenyl(114C)methanol Chemical compound O[14CH2]C1=CC=CC=C1 WVDDGKGOMKODPV-ZQBYOMGUSA-N 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- 230000006337 proteolytic cleavage Effects 0.000 description 1
- 125000003226 pyrazolyl group Chemical group 0.000 description 1
- 125000004076 pyridyl group Chemical group 0.000 description 1
- LDNSHTVNGGHGQJ-UHFFFAOYSA-N pyrrole-1-carboxamide Chemical compound NC(=O)N1C=CC=C1 LDNSHTVNGGHGQJ-UHFFFAOYSA-N 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 1
- LOAUVZALPPNFOQ-UHFFFAOYSA-N quinaldic acid Chemical compound C1=CC=CC2=NC(C(=O)O)=CC=C21 LOAUVZALPPNFOQ-UHFFFAOYSA-N 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- RWWYLEGWBNMMLJ-MEUHYHILSA-N remdesivir Drugs C([C@@H]1[C@H]([C@@H](O)[C@@](C#N)(O1)C=1N2N=CN=C(N)C2=CC=1)O)OP(=O)(N[C@@H](C)C(=O)OCC(CC)CC)OC1=CC=CC=C1 RWWYLEGWBNMMLJ-MEUHYHILSA-N 0.000 description 1
- RWWYLEGWBNMMLJ-YSOARWBDSA-N remdesivir Chemical compound NC1=NC=NN2C1=CC=C2[C@]1([C@@H]([C@@H]([C@H](O1)CO[P@](=O)(OC1=CC=CC=C1)N[C@H](C(=O)OCC(CC)CC)C)O)O)C#N RWWYLEGWBNMMLJ-YSOARWBDSA-N 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 239000008281 solid sol Substances 0.000 description 1
- 238000009987 spinning Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 229940095064 tartrate Drugs 0.000 description 1
- FAGLEPBREOXSAC-UHFFFAOYSA-N tert-butyl isocyanide Chemical compound CC(C)(C)[N+]#[C-] FAGLEPBREOXSAC-UHFFFAOYSA-N 0.000 description 1
- 230000008719 thickening Effects 0.000 description 1
- 125000001544 thienyl group Chemical group 0.000 description 1
- 231100000820 toxicity test Toxicity 0.000 description 1
- 229910052722 tritium Inorganic materials 0.000 description 1
- 229960003732 tyramine Drugs 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- NQPDZGIKBAWPEJ-UHFFFAOYSA-N valeric acid Chemical compound CCCCC(O)=O NQPDZGIKBAWPEJ-UHFFFAOYSA-N 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 239000008096 xylene Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K31/4025—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil not condensed and containing further heterocyclic rings, e.g. cromakalim
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/42—Oxazoles
- A61K31/422—Oxazoles not condensed and containing further heterocyclic rings
-
- 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
- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
- C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D207/16—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
Abstract
Description
本申请是对申请号为2020115682827,申请日为2020年12月25日的发明专利提出的分案申请。
技术领域
本发明属于有机合成药物技术领域,具体涉及一种新型冠状病毒主蛋白酶的抑制剂及其制备方法和制药用途。
背景技术
冠状病毒的基因组RNA长约30knt,具有5′帽结构和3′-poly-a尾,至少含有6个开放阅读框(ORF)。第一个ORF(ORF1a/b)约占基因组长度的三分之二,直接翻译两种多蛋白:pp1a和pp1ab,ORF1a和ORF1b之间存在a-1移码。这些多蛋白由一种主蛋白酶(简称Mpro;也被称为3C样蛋白酶(3CLpro))和一个或两个木瓜蛋白酶样蛋白酶(PLPs)加工而成,转化为16种非结构蛋白。这些非结构蛋白参与亚基因组RNA的生产,编码四种主要结构蛋白(包膜(E)、膜(M)、棘突(S)和核衣壳(N)蛋白质)和其他辅助蛋白质,以完成病毒的复制和侵入过程。
Mpro将重叠的pp1a和pp1ab多聚蛋白水解裂解为功能蛋白,这是病毒复制过程中的关键步骤。对于RdRp或nsp13等病毒复制必需的酶,如果没有事先的蛋白水解释放,就不能完全发挥作用完成复制。因此,抑制病毒的Mpro可以阻止传染性病毒颗粒的产生,从而减轻疾病症状。
Mpro在冠状病毒中是保守的,并且不同冠状病毒中Mpro的底物具有一些共同的特征:从N端到C端的氨基酸以配对的形式进行编号(-P4-P3-P2-P1↓P1′-P2′-P3′),裂解位点在P1和P1′之间。特别地,Mpro在P1位点(Leu-Gln↓(Ser,Ala,Gly))对谷氨酰胺有独特的底物偏好,这一点在宿主蛋白酶中是不存在的,这表明通过靶向病毒Mpro实现高选择性是可行的。因此,病毒对这种蛋白酶的正确功能的绝对依赖性,加上缺乏同源的人类蛋白酶,使得Mpro成为理想的抗病毒靶点。
因此,亟需研究出一种能够有效抑制SARS-CoV-2病毒的Mpro活性的药物。
发明内容
本发明的目的是提供新型冠状病毒主蛋白酶的抑制剂及其制备方法和制药用途。
本发明提供了式I所示化合物、或其药学上可接受的盐、或其立体异构体、或其旋光异构体、或其同位素替代形式:
其中,X为O或S;
A环选自未取代或被一个或多个R6取代的以下基团:5~6元饱和杂环基、5~6元不饱和杂环基、饱和杂稠环基、不饱和杂稠环基;R6各自独立的选自C1~6烷基、C1~6烷氧基、卤素、羟基、氰基、氨基、羧基;
R3为L3M0L4R3a;其中L3选自无、C1~4亚烷基、卤代C1~4亚烷基、C2~4亚烯基、卤代C2~4亚烯基,L4选自无、C1~4亚烷基、卤代C1~4亚烷基,M0选自无、O、S、NH、CO、CONH、NHCO,R3a为未取代或被一个或多个R3b取代的以下基团:5~6元芳基、5~6元杂芳基、不饱和杂稠环基、不饱和稠环烷基;R3b各自独立的选自被R3c取代或未取代的C1~5烷基、被R3c取代或未取代的C1~5烷氧基、卤素、被R3c取代或未取代的苯基、NR14R15、被R3c取代或未取代的萘基、羟基;R14、R15各自独立的选自氢或C1~5烷基,R3c各自独立的选自卤素、氘、氰基、羟基、氨基、羧基;
R4选自未取代或被一个或多个取代基取代的以下基团:5~6元芳基、5~6元杂芳基、C1~5烷基、COOR10;所述取代基各自独立的选自=O、羟基、硝基、氨基、羧基、卤素、C1~5烷基;R10为C1~5烷基;
R5选自COR8或WCOOR7;其中,R8选自氢或W选自无、C1~4亚烷基、C2~4亚烯基、C2~4亚炔基,R7选自C1~6烷基;M选自无、CO、NH、CONH、NHCO、COO或OCO,L0选自无、C1~4亚烷基、C2~4亚烯基,L1选自无、C1~4亚烷基、C2~4亚烯基,R8a选自C1~5烷基、卤代的C1~5烷基、3~6元饱和环烷基、3~6元饱和杂环基、5~6元芳基或5~6元杂芳基。
进一步地,所述化合物的结构如式II、式III或式IV所示:
其中,X为O或S;
n选自0~3的整数,优选为0~2的整数;
R1、R2各自独立的选自氢、C1~5烷基、C1~5烷氧基、卤素、羟基、氰基、氨基、羧基;
R3为L3M0L4R3a;其中L3选自无、C1~4亚烷基、卤代C1~4亚烷基、C2~3亚烯基,L4选自无、C1~4亚烷基、卤代C1~4亚烷基,M0选自无、O、S、NH、CO、CONH、NHCO,R3a为未取代或被一个或多个R3b取代的以下基团:苯基、 R3b各自独立的选自C1~4烷基、卤素取代的C1~4烷基、氘代的C1~4烷基、氰基取代的C1~4烷基、C1~4烷氧基、卤素取代的C1~4烷氧基、氘代的C1~4烷氧基、氰基取代的C1~4烷氧基、卤素、苯基、卤代的苯基、NR14R15、羟基,R14、R15各自独立的选自氢或C1~4烷基;
R4选自未取代或被一个或多个取代基取代的以下基团:5~6元芳基、5~6元杂芳基、C1~5烷基、COOR10;所述取代基各自独立的选自=O、羟基、硝基、氨基、羧基、卤素、C1~5烷基;R10为C1~5烷基;
R8选自氢或M选自无、CO、NH、CONH、NHCO、COO或OCO,L0选自无、C1~3亚烷基、C2~4亚烯基,L1选自无、C1~3亚烷基、C2~4亚烯基,R8a选自C1~4烷基、卤代的C1~4烷基、3~6元饱和环烷基、3~6元饱和杂环基、5~6元芳基或5~6元杂芳基。
进一步地,R1、R2各自独立的选自氢、C1~4烷基、C1~4烷氧基、卤素、羟基;
R3选自 L3M0L4R3a;L3选自无、C1~3亚烷基、卤代C1~3亚烷基、C2~3亚烯基,L4选自无、C1~3亚烷基、卤代C1~3亚烷基、,M0选自无、O、NH、CO、CONH,R3a为苯基、被一个或多个R3b取代的苯基,R3b各自独立的选自C1~4烷基、卤素取代的C1~4烷基、氘代的C1~4烷基、氰基取代的C1~4烷基、C1~4烷氧基、卤素取代的C1~4烷氧基、氘代的C1~4烷氧基、氰基取代的C1~4烷氧基、卤素、苯基、卤代的苯基、NR14R15、羟基,R14、R15各自独立的选自氢或C1~3烷基;
R4选自C1~2烷基、COOR10、取代或未取代的苯基;所述取代基选自羟基、硝基;Ra1、Ra2各自独立的选自氢、C1~3烷基、卤素;R10为C1~3烷基;
R8选自氢、CONHR11、L2COOR12、C1~4烷基、卤代的C1~4烷基;R11选自3~6元饱和环烷基、C1~4烷基、苄基、L2为C1~2亚烷基、C2~3亚烯基,R12为C1~3烷基。
进一步地,所述式II如式II-1或式II-2所示:
其中,X为O或S,优选为O;
R1、R2各自独立的选自氢、C1~3烷基,优选为甲基;
m选自0~3的整数,R3b各自独立的选自苯基、卤代的苯基、卤素、C1~3烷基、卤代或氘代的C1~3烷基、C1~3烷氧基、卤代或氘代的C1~3烷氧基、羟基;
Ra1、Ra2各自独立的选自氢、C1~3烷基、卤素;
Rb选自氢、C1~3烷基、卤代的C1~3烷基;
L3选自无、C1~2亚烷基、卤代C1~2亚烷基、C2亚烯基,L4选自无、C1~3亚烷基、卤代C1~3亚烷基,M0选自无、O、NH、CO、CONH;
所述卤素优选为氯、氟。
进一步地,所述化合物的结构为以下结构之一:
本发明还提供了一种药物组合物,所述药物组合物是以上述化合物、或其药学上可接受的盐、或其立体异构体、或其旋光异构体、或其同位素替代形式为活性成分,加上药学上可接受的辅料制成的制剂。
本发明还提供了上述化合物、或其药学上可接受的盐、或其立体异构体、或其旋光异构体、或其同位素替代形式在制备冠状病毒蛋白水解酶抑制剂中的用途;优选的,所述冠状病毒蛋白水解酶为冠状病毒主蛋白酶;更优选的,所述冠状病毒蛋白水解酶为SARS-COV-2Mpro。
本发明还提供了上述化合物、或其药学上可接受的盐、或其立体异构体、或其旋光异构体、或其同位素替代形式在制备抗冠状病毒的药物中的用途,优选的,所述冠状病毒为新型冠状病毒SARS-CoV-2。
本发明还提供了上述化合物、或其药学上可接受的盐、或其立体异构体、或其旋光异构体、或其同位素替代形式在制备预防和/或治疗与SARS-COV-2Mpro相关的疾病的药物中的用途,优选的,所述与SARS-COV-2Mpro相关的疾病为新型冠状病毒COVID-19。
进一步地,所述冠状病毒蛋白水解酶抑制剂、抗冠状病毒的药物或预防和/或治疗病毒性肺炎的药物能够抑制SARS-COV-2Mpro的活性和/或能够抑制SARS-COV-2感染细胞。
关于本发明的使用术语的定义:除非另有说明,本文中基团或者术语提供的初始定义适用于整篇说明书的该基团或者术语;对于本文没有具体定义的术语,应该根据公开内容和上下文,给出本领域技术人员能够给予它们的含义。
碳氢基团中碳原子含量的最小值和最大值通过前缀表示,例如,前缀Ca~b烷基表示任何含“a”至“b”个碳原子的烷基。例如,C1~6烷基是指包含1~6个碳原子的直链或支链的烷基。
本文“取代”是指分子中的1个、2个或多个氢原子被其它不同的原子或分子所替换,包括该分子中同位原子或异位原子上的1个、2个或多个取代。
“同位素替代形式”指化合物中的一个或两个以上的原子被其对应的同位素替换后得到的化合物,比如化合物中的氢被替换为氕、氘或氚。
“药学上可接受的”是指某载体、运载物、稀释剂、辅料,和/或所形成的盐通常在化学上或物理上与构成某药物剂型的其它成分相兼容,并在生理上与受体相兼容。
“盐”是将化合物或其立体异构体,与无机和/或有机酸和/或碱形成的酸式和/或碱式盐,也包括两性离子盐(内盐),还包括季铵盐,例如烷基铵盐。这些盐可以是在化合物的最后分离和纯化中直接得到。也可以是通过将化合物,或其立体异构体,与一定数量的酸或碱适当(例如等当量)进行混合而得到。这些盐可能在溶液中形成沉淀而以过滤方法收集,或在溶剂蒸发后回收而得到,或在水介质中反应后冷冻干燥制得。
“药学上可接受的盐”可以是化合物的盐酸盐、硫酸盐、枸橼酸盐、苯磺酸盐、氢溴酸盐、氢氟酸盐、磷酸盐、乙酸盐、丙酸盐、丁二酸盐、草酸盐、苹果酸盐、琥珀酸盐、富马酸盐、马来酸盐、酒石酸盐或三氟乙酸盐。
卤素为氟、氯、溴或碘。
“芳基”指具有共轭的π电子体系的全碳单环或稠合多环(也就是共享毗邻碳原子对的环)基团,例如苯基。所述芳基不含有杂原子,如氮,氧,或硫,同时连接母体的点必须在具有共轭的π电子体系的环上的碳原子上。芳基可以是取代的或未取代的。“5~6元芳基”指环碳原子数为5或6的芳基。
“杂芳基”指包含一个到多个杂原子的杂芳族基团。这里所指的杂原子包括氧、硫和氮。例如呋喃基、噻吩基、吡啶基、吡唑基等。所述杂芳基可以是任选取代的或未取代的。“5~6元杂芳基”指环原子数为5或6的杂芳基。
“环烷基”指饱和或不饱和的环状烃取代基;环状烃可以是单环也可以是多环。例如,“3~6元饱和环烷基”指环碳原子数为3~6的饱和的环烷基。
“杂环基”指饱和或不饱和的环状烃取代基;环状烃可以是单环也可以是多环,且携带至少一个环杂原子(包括但不限于O、S或N)。例如,“3~6元饱和杂环基”指环原子数为3~6的饱和的杂环基。
“稠环烷基”指多环的环烷基,且该多环的环烷基中有两个环共用两个相邻的碳原子。
“杂稠环基”指多环的杂环基,其中至少含有1个杂原子,且该多环的杂环基中有两个环共用两个相邻的碳原子或杂原子。
“亚烷基”指烷基失去一个原子后的基团。例如C1亚烷基:C2亚烷基:
“亚烯基”指烯基失去一个原子后的基团。例如C2烯基:
“亚炔基”指炔基失去一个原子后的基团。例如C2炔基:
实验结果表明,本发明提供了一种能够有效抑制新型冠状病毒主蛋白酶Mpro活性的化合物,该化合物能够有效抑制SARS-COV-2病毒在细胞内的复制,抑制细胞中的SARS-COV-2感染,抵抗转基因小鼠的体内SARS-COV-2感染;降低SARS-COV-2感染的转基因小鼠肺部的病毒载量,降低小鼠肺部趋化因子配体10(CXCL10)和β型干扰素(IFN-β)的基因表达水平,降低小鼠肺部的中性粒细胞(NEU)和巨噬细胞(MAC)的数量,改善小鼠肺部的病理损伤。同时,本发明提供的化合物还具有良好的体内安全性和药代动力学性质。本发明的化合物在制备SARS-CoV-2Mpro抑制剂,抗SARS-CoV-2的药物,以及预防和/或治疗新型冠状病毒的药物中具有非常好的应用前景。
显然,根据本发明的上述内容,按照本领域的普通技术知识和惯用手段,在不脱离本发明上述基本技术思想前提下,还可以做出其它多种形式的修改、替换或变更。
以下通过实施例形式的具体实施方式,对本发明的上述内容再作进一步的详细说明。但不应将此理解为本发明上述主题的范围仅限于以下的实例。凡基于本发明上述内容所实现的技术均属于本发明的范围。
附图说明
图1为化合物26对SARS-COV-2Mpro的抑制活性曲线。
图2为化合物33对SARS-COV-2Mpro的抑制活性曲线。
图3为化合物37对SARS-COV-2Mpro的抑制活性曲线。
图4化合物对SARS-COV-2在人肺泡上皮细胞中复制的抑制实验。
图5SARS-CoV-2感染小鼠的肺部病毒载量。
图6SARS-CoV-2感染小鼠的肺部病理组织切片(3dpi)。
图7SARS-CoV-2感染小鼠的肺部代表性细胞因子表达水平(3dpi)。
图8SARS-CoV-2感染小鼠的肺部中性粒细胞和巨噬细胞计数(3dpi)。
具体实施方式
本发明所用原料与设备均为已知产品,通过购买市售产品所得。
实施例1:化合物1的制备
按照上述制备路线制得本发明的化合物1,路线中各步骤的反应条件如下:
i、a,2-氟丙二酸二甲酯,苄醇,甲苯,对甲苯磺酸,110℃;b,异丙醇,正己烷,-10℃;
ii、异丙醇,氢氧化钠,水,45℃;
iii、无水四氢呋喃,异丙基氯化镁四氢呋喃溶液,Ar,0℃;
iv、无水四氢呋喃,N,N'-羰基二咪唑,Ar,0℃;
v、乙酸乙酯,10%钯碳,氢气,室温;
vi、二氯甲烷,盐酸二氧六环溶液;
vii、二氯甲烷,N,N,N′,N′-四甲基-O-(7-氮杂苯并三唑-1-基)六氟磷酸脲,N,N-二异丙基乙胺,-20℃;
viii、二氯甲烷,三氟乙酸;
ix、二氯甲烷,N,N,N′,N′-四甲基-O-(7-氮杂苯并三唑-1-基)六氟磷酸脲,N,N-二异丙基乙胺,-20℃;
以下为具体合成步骤:
中间体2:2-氟丙二酸二苄酯的制备
2-氟丙二酸二甲酯(10g,66.6mmol,1.0eq)和苄醇(35mL,338.2mmol,5.0eq)用100mL甲苯溶解,加入1.15g对甲苯磺酸(6.7mmol,0.1eq),回流反应,TLC监控反应,约8小时后反应完毕。冷却至室温,减压蒸去甲苯,加入15mL异丙醇,搅拌均匀,搅拌下慢慢加入30mL正己烷,置于-10℃冷阱中继续搅拌2小时,析出大量白色固体。抽滤,滤饼用10mL×2冷冻正己烷洗涤两次,滤饼30℃真空减压干燥得18.4g产品,收率91.4%。1H NMR(400MHz,DMSO-d6)δ7.64–6.91(m,10H),6.00(d,J=46.3Hz,1H),5.30–5.20(m,4H).
中间体3:2-氟丙二酸单苄酯的制备
2-氟丙二酸二苄酯(18.4g,60.9mmol,1.0eq)用100mL异丙醇溶解,升温至45℃,氢氧化钠(2.55g,63.9mmol,1.05eq)溶于60mL水后慢慢滴入,滴加时间>1小时。滴加完毕后继续反应30分钟,减压蒸去异丙醇,加入50mL水,用饱和碳酸氢钠溶液调节pH值至9左右。水相用二氯甲烷20mL×2二氯甲烷萃取两次,用6mol/L盐酸调节水相pH值至1-2,用40mL×3异丙醚萃取三次,合并有机相,用30mL饱和盐水洗涤一次。有机相加入无水硫酸镁干燥,过滤,浓缩得粘稠状残留物,加入60mL正己烷搅拌过夜,析出白色固体,过滤,滤饼40℃真空减压干燥得6.5g产品,收率50.3%.1H NMR(400MHz,Chloroform-d)δ7.41–7.32(m,5H),5.87(s,2H),5.39(d,J=47.9Hz,1H),5.31(s,1H).
中间体4的制备
2-氟丙二酸单苄酯溶于无水四氢呋喃(2mL/mmol),氩气置换保护,冷却至0℃,慢慢滴加异丙基氯化镁四氢呋喃溶液(2M四氢呋喃溶液,2.0eq),得白色悬浊液。0℃下继续搅拌1小时,产品悬浊液直接用于下步反应。
中间体6:1-苄基6-甲基(4S)-4-(((叔丁氧羰基)氨基)-2-氟-3-氧代己二酸酯的制备
Boc-L-天冬氨酸4-甲酯(2.2g,8.8mmol,1.0eq)溶于50mL无水四氢呋喃,氩气置换保护,冷却至0℃,加入CDI(1.5g,9.3mmol,1.05eq),保温反应1小时。反应液冷却至-20℃,慢慢加入1.5eq中间体4,保温反应1小时后升温至室温反应6小时。冰水浴下将反应液慢慢倒入300mL 2M稀盐酸中,用100mL×3乙酸乙酯萃取三次,合并有机相,用饱和碳酸氢钠溶液洗涤至弱碱性,再用50mL饱和盐水洗涤一次后加入无水硫酸镁干燥,过滤,浓缩,所得粗品直接用于下步反应。
中间体7:(S)-3-(((叔丁氧羰基)氨基)甲基-5-氟-4-氧戊酸甲酯的制备
上步所得中间体6粗品加入50mL乙酸乙酯,加入200mg 10%钯碳,氢气置换,氢气下室温反应过夜,过滤,浓缩,所得粗品用石油醚:乙酸乙酯=10:1流动相柱层析得1.5g无色油状物,收率65%。1H NMR(400MHz,Chloroform-d)δ5.51(d,J=8.0Hz,1H),5.28–5.06(m,2H),4.73–4.52(m,1H),3.70(s,3H),3.08(dd,J=17.2,4.6Hz,1H),2.84(dd,J=17.2,5.0Hz,1H),1.46(s,9H).
中间体8:(S)-3-氨基-5-氟-4-氧戊酸甲酯的制备
500mg中间体7加入5mL二氯甲烷溶解,然后加入5mL盐酸二氧六环,反应完全后旋干,得中间体8,收率为91.2%。1H NMR(400MHz,Chloroform-d)δ5.25-5.10(m,2H),4.53(dd,J=8.7,1.0Hz,2H),4.44(d,J=7.9Hz,1H),3.69(s,3H),2.83–2.71(m,2H).
中间体11:甲基(1H-吲哚-2-羰基)-L-脯氨酸的制备
1H-吲哚-2-羧酸(1g,6.21mmol,1.0eq)用二氯甲烷溶解后,于-20℃加入HATU(2.81g,7.40mmol,1.2eq)后加入L-脯氨酸甲酯盐酸盐(1.03g,6.21mmol,1.0eq),最后加入DIEA(3mL,18.51mmol,3.0eq),TLC监控反应。反应完毕后,用水溶液和DCM进行萃取,有机层浓缩后经柱层析分离得到中间体11(1.53g),收率为75.2%。1H NMR(400MHz,DMSO-d6)δ7.68(dt,J=7.4,1.5Hz,1H),7.43(dd,J=7.4,1.6Hz,1H),7.26(td,J=7.5,1.7Hz,1H),7.19–7.14(m,2H),4.31(t,J=7.0Hz,1H),3.72(td,J=7.1,2.3Hz,2H),3.68(s,3H),2.11–2.00(m,2H),1.93–1.81(m,2H)。
中间体12:(1H-吲哚-2-羰基)-L-脯氨酸的制备
500mg中间体11加入10mL二氯甲烷溶解,然后加入5mL三氟乙酸,反应完全后旋干,得378mg中间体12,直接作为先一步反应。收率为91.2%。
化合物1:甲基(S)-3-((S)-1-(1H-吲哚-2-羰基)吡咯烷-2-甲酰胺)-5-氟-4-氧代戊酸的制备
中间体12(168mg,0.61mmol,1.0eq)用二氯甲烷溶解后,于-20℃加入HATU(280mg,0.73mmol,1.2eq)后加入中间体8(100mg,0.61mmol,1.0eq),最后加入DIEA(301μL,1.83mmol,3.0eq),TLC监控反应。反应完毕后,用水溶液和DCM进行萃取,浓缩有机层,柱层析分离得到化合物1,收率为34%。1H NMR(400MHz,DMSO)δ11.55(s,1H),8.69(s,1H),7.65(d,J=7.6Hz,1H),7.46(d,J=8.3Hz,1H),7.20(m,1H),7.06(d,J=7.8Hz,2H),5.26(m,2H),4.60(m,1H),4.49(m,1H),3.96(dd,J=15.0,7.4Hz,2H),3.61(s,3H),2.86(m,1H),2.60(dd,J=15.9,7.7Hz,1H),2.02(m,2H),1.82(m,2H).HRMS m/z(ESI)calcd forC20H25FN4O5[M+H]+403.1543found:404.1476。
实施例2:化合物3的制备
按照上述制备路线制得本发明的化合物3,路线中各步骤的反应条件如下:
i、Boc-L-谷氨酸二甲酯,LiHMDS四氢呋喃溶液,氩气,无水四氢呋喃,-78℃,;
ii、(2S,4R)-二甲基2-(叔-丁氧基羰基氨基)-4-(氰基甲基)戊二酸酯,无水甲醇,六水氯化钴,硼氢化钠;
iii、(S)-甲基2-(叔-丁氧基羰基氨基)-3-((S)-2-羰基吡咯烷-3-基)丙酸酯,一水合氢氧化锂,四氢呋喃,0℃;
iv、无水四氢呋喃,Ar,N,N'-羰基二咪唑,0℃;
v、乙酸乙酯,10%钯碳,氢气,室温;
vi、二氯甲烷,盐酸二氧六环溶液;
vii、二氯甲烷,N,N,N′,N′-四甲基-O-(7-氮杂苯并三唑-1-基)六氟磷酸脲,N,N-二异丙基乙胺,-20℃;
viii、二氯甲烷,三氟乙酸;
ix、二氯甲烷,N,N,N′,N′-四甲基-O-(7-氮杂苯并三唑-1-基)六氟磷酸脲,N,N-二异丙基乙胺,-20℃;
以下为具体合成步骤:
中间体14:(2S,4R)-2-((叔丁氧羰基)氨基)-4-(氰基甲基)戊二酸二甲酯的制备
Boc-L-谷氨酸二甲酯(12g,43.6mmol,1.0eq)溶于100mL无水四氢呋喃,氩气置换保护,冷却至-78℃,慢慢滴加94mL LiHMDS四氢呋喃溶液(1M四氢呋喃溶液,94mmol,2.2eq),滴加完毕后保温反应1小时。3.24mL溴乙腈(46.6mmol,1.1eq)慢慢滴加入反应液,保温反应6小时后用50mL饱和氯化铵溶液淬灭反应。将淬灭后的反应液升温至室温,用60mL×3乙酸乙酯萃取三次,合并有机相,用50mL饱和盐水洗涤后加入无水硫酸镁干燥,过滤,浓缩,所得粗品用石油醚:乙酸乙酯=4:1流动相柱层析得9.36g浅黄色油状物,收率68.3%。1H NMR(400MHz,Chloroform-d)δ5.11(d,J=8.6Hz,1H),4.39(s,1H),3.77(s,3H),3.76(s,3H),2.90–2.82(m,1H),2.82–2.74(m,2H),2.28–2.06(m,2H),1.45(s,9H).
中间体15:(S)-2-(((叔丁氧羰基)氨基)甲基-3-((S)-2-氧吡咯烷-3-基)丙酸甲酯的制备
(2S,4R)-二甲基2-(叔-丁氧基羰基氨基)-4-(氰基甲基)戊二酸酯(9.36g,29.8mmol,1.0eq)溶于150mL无水甲醇,冷却至0℃。加入六水氯化钴(4.25g,18mmol,0.6eq),再分批加入硼氢化钠(6.76g,180mmol,6.0eq),加完后升温至室温,反应过夜,TLC监控反应完毕。加入50mL饱和氯化铵溶液淬灭反应,减压蒸去甲醇,用100mL×3乙酸乙酯萃取三次,合并有机相,依次用200mL×3饱和氯化铵溶液洗涤三次、200mL×3饱和盐水洗涤三次,有机相加入无水硫酸镁干燥,过滤,浓缩,所得粗品用石油醚:乙酸乙酯=1:1流动相柱层析得3.94g白色固体,收率46.2%。1H NMR(400MHz,Chloroform-d)δ5.92(s,1H),5.49(d,J=8.4Hz,1H),4.41–4.26(m,1H),3.74(s,3H),3.45–3.26(m,2H),2.58–2.39(m,2H),2.27–2.07(m,1H),1.98–1.78(m,2H),1.44(s,9H).
中间体16:(S)-2-((叔丁氧羰基)氨基)-3-((S)-2-氧吡咯烷-3-基)丙酸的制备
(S)-甲基2-(叔-丁氧基羰基氨基)-3-((S)-2-羰基吡咯烷-3-基)丙酸酯(0.88g,3.1mmol,1.0eq)溶于10mL四氢呋喃,冷却至0℃。一水合氢氧化锂(0.64g,15.4mmol,5.0eq)溶于10mL水后慢慢滴入,滴完后保温反应4小时,TLC监控反应完毕。饱和柠檬酸水溶液调节pH值至中性,减压蒸去四氢呋喃,10mL乙酸乙酯萃取一次,水相用饱和柠檬酸水溶液调节pH值至3-4,20mL×3乙酸乙酯萃取三次,合并有机相,用20mL饱和盐水洗涤后加入无水硫酸镁干燥,过滤,浓缩得0.78g类白色固体,收率93.2%。1H NMR(400MHz,Chloroform-d)δ7.19(s,1H),5.69(d,J=7.9Hz,1H),4.35(q,J=7.6Hz,1H),3.48–3.31(m,2H),2.70–2.55(m,1H),2.51–2.36(m,1H),2.27–2.12(m,1H),1.99–1.80(m,2H),1.44(s,9H).
中间体17:(4S)-4-((叔丁氧羰基)氨基)-2-氟-3-氧代-5-((S)-2-氧吡咯烷-3-基)戊酸苄酯的制备
(S)-2-((叔-丁氧基羰基)氨基)-3-((S)-2-羰基吡咯烷-3-基)丙酸(2.4g,8.8mmol,1.0eq)溶于50mL无水四氢呋喃,氩气置换保护,冷却至0℃,加入CDI(1.5g,9.3mmol,1.05eq),保温反应1小时。反应液冷却至-20℃,慢慢加入1.5eq中间体4,保温反应1小时后升温至室温反应6小时。冰水浴下将反应液慢慢倒入300mL 2M稀盐酸中,用100mL×3乙酸乙酯萃取三次,合并有机相,用饱和碳酸氢钠溶液洗涤至弱碱性,再用50mL饱和盐水洗涤一次后加入无水硫酸镁干燥,过滤,浓缩,所得粗品直接用于下步反应。
中间体18:((S)-4-氟-3-氧代-1-((S)-2-氧吡咯烷-3-基)丁烷-2-基)氨基甲酸叔丁酯的制备
上步所得中间体17粗品加入50mL乙酸乙酯,加入200mg 10%钯碳,氢气置换,氢气下室温反应过夜,过滤,浓缩,所得粗品用石油醚:乙酸乙酯=1:1流动相柱层析得1.3g白色固体,收率50%。1H NMR(400MHz,Chloroform-d)δ5.99(d,J=7.5Hz,1H),5.91(s,1H),5.31–4.95(m,2H),4.56(s,1H),3.42–3.32(m,2H),2.56–2.42(m,2H),2.10–1.97(m,1H),1.96–1.81(m,2H),1.45(s,9H).
中间体19:(S)-3-((S)-2-氨基-4-氟-3-氧丁基)吡咯烷-2-酮的制备
500mg中间体18加入5mL二氯甲烷溶解,然后加入5mL盐酸二氧六环,反应完全后旋干,得中间体19,收率为85%。
中间体22:乙基(1S,3aR,6aS)-2-(2-(2,4-二氯苯氧)乙酰基)八氢环戊烯并[c]吡咯-1-羧酸酯的制备.
将2,4-二氯苯氧乙酸(中间体21,0.58g,2.62mmol),2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(1.2g,3.14mmol),N,N-二异丙基乙胺(1.3mL,7.86mmol),and(1S,3aR,6aS)-八氢环戊烯并[c]吡咯-1-羧酸乙酯盐酸盐(中间体20,0.58g,2.62mmol)溶于15mL超干N,N-二甲基甲酰胺中,反应在25℃氩气保护条件下反应12小时,d反应加入4倍体积水,用二氯甲烷萃取三次,合并有机相用饱和氯化铵溶液、饱和碳酸钠溶液洗涤,无水硫酸钠干燥后过滤,硅胶拌样柱层析(石油醚/乙酸乙酯=1:1)得到中间体22(0.60g,59%)。1H NMR(400MHz,MeOD)δ7.42(d,J=5.4Hz,1H),7.26–7.19(m,1H),6.97(d,J=8.9Hz,1H),4.80-7.72(m,2H),4.28(d,J=3.6Hz,1H),4.22–4.10(m,2H),3.87(d,J=10.6Hz,1H),3.63–3.48(m,1H),3.57(d,J=10.5Hz,2H),2.71–2.61(m,1H),2.08–1.84(m,1H),1.83–1.46(m,4H),1.31–1.17(m,3H).ESI-MS(m/z):386.02(M+H)+.
中间体23:(1S,3aR,6aS)-2-(2-(2,4-二氯苯氧)乙酰基)八氢环戊烯并[c]吡咯-1-羧酸的制备
将乙基(1S,3aR,6aS)-2-(2-(2,4-二氯苯氧)乙酰基)八氢环戊烯并[c]吡咯-1-羧酸酯(中间体22,200mg,0.52mmol)溶于20mL甲醇中,然后加入2M氢氧化钠溶液(10mL),反应在25℃下搅拌4小时.TLC监控反应结束后,旋干甲醇,用盐酸调pH至弱酸性,二氯甲烷萃取三次,合并有机相无水硫酸钠干燥后选干得粗产品直接进行下一步反应。
化合物3:(1S,3aR,6aS)-2-(2-(2,4-二氯)乙酰基)-N-((S)-4-氟-3-氧-1-((S)-2-羰基-3-基)丁烷-2-基)八氢环戊烯并[c]吡咯-1-甲酰胺的制备
中间体23(168mg,0.61mmol,1.0eq)用二氯甲烷溶解后,于-20℃加入HATU(280mg,0.73mmol,1.2eq)后加入中间体19(100mg,0.61mmol,1.0eq),最后加入DIEA(301μL,1.83mmol,3.0eq),TLC监控反应。反应完毕后,用水溶液和DCM进行萃取,有机层浓缩后经柱层析分离得到化合物3,收率为34%。1H NMR(400MHz,DMSO)δ8.61(d,J=7.4Hz,2H),8.29(d,J=7.7Hz,1H),8.15(d,J=8.5Hz,1H),7.65(s,1H),7.56(d,J=7.0Hz,2H),7.41(td,J=11.1,6.0Hz,4H),6.75(dd,J=15.9,6.1Hz,1H),5.15(m,2H),4.39(s,1H),3.62(d,J=4.1Hz,2H),3.16(m,1H),3.11(m,2H),2.28(d,J=36.4Hz,1H),2.12(s,1H),1.96(m,1H),1.62(m,2H),1.50(dd,J=15.5,8.6Hz,2H),0.88(m,6H).HRMS m/z(ESI)calcd forC23H30FN3O4[M+H]+432.2293found:432.2291。
实施例3:制备化合物9
按照上述制备路线制得本发明的化合物9,路线中各步骤的反应条件如下:
i、1-羟基苯并三唑,1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐,N,N-二异丙基乙胺,N,N-二甲基甲酰胺,室温。
ii、氢氧化钠,甲醇,水,55度
iii、2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯,N,N-二异丙基乙胺,N,N-二甲基甲酰胺,0℃
iv、硼氢化钠,甲醇,室温
v、戴斯马丁氧化剂,超干二氯甲烷,室温
以下为具体合成步骤:
中间体25:甲基(1R,2S,5S)-6,6-二甲基3-(喹啉-2-羰基)-3-氮杂双环[3.1.0]己烷-2-羧酸的制备
将原料23(喹啉2-羧酸1.0g,11.6mmol)、1-羟基苯并三唑(2.03g,15.08mmoL)、1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(4.43g,23.3mmol),N,N-二甲基甲酰胺30mL置于圆底烧瓶常温搅拌0.5小时,加入N,N-二异丙基乙胺2.5mL,再加入0.59g中间体24,反应8h后,减压蒸馏除去溶剂,用二氯甲烷和氯化铵溶液、碳酸氢钠溶液进行萃取,水、饱和氯化钠溶液进行洗涤,最后用硫酸钠干燥,抽滤,将有机相柱层析得白色固体。收率为85%。1HNMR(400MHz,DMSO)δ8.12–8.03(m,1H),8.00–7.90(m,2H),7.66–7.48(m,3H),4.54(s,1H),4.03(q,J=7.1Hz,1H),3.75(d,J=6.7Hz,3H),3.47(d,J=5.4Hz 1H),3.39(d,J=5.7Hz1H),1.99(t,J=6.2Hz,1H),1.54(t,J=6.8Hz,1H),1.03(s,3H),0.97(s,3H).MS(ESI,正离子)m/z:325.04.87[M+H]+。
中间体26:(1R,2S,5S)-6,6-二甲基3-(喹啉-2-羰基)-3-氮杂双环[3.1.0]己烷-2-羧酸的制备
将中间体25溶于20mL四氢呋喃,缓慢加入10mL 2mol/L的氢氧化钠溶液,将反应液在逐渐升温至55度搅拌3小时后终止并冷却至常温,浓缩反应液,加水调pH为弱酸性,析出白色固体,抽滤的得中间体5,未经进一步纯化被用作下一步反应
中间体28:甲基((1R,5S)-6,6-二甲基-3-(喹啉-2-羰基)-3-氮杂双环[3.1.0]己烷-2-羰基)-L-苯基丙氨酸的制备
将中间体26 1.0g、2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯1.93g加入20mL N,N-二甲基甲酰胺中,0℃搅拌0.5小时,加入N,N-二异丙基乙胺2.0mL,再加入中间体27 0.89g,氩气保护0℃条件下反应12h后,加入4倍体积的水,用乙酸乙酯进行萃取三次。合并有机相,用氯化铵溶液、碳酸氢钠溶液进行萃取,水、饱和氯化钠溶液进行洗涤,最后用硫酸钠干燥,抽滤,将有机相柱层析得白色固体。收率为65%。1H NMR(400MHz,DMSO)δ8.53(d,J=7.5Hz,1H),8.04(d,J=6.4Hz,1H),7.99(d,J=6.4Hz,1H),7.94(d,J=3.7Hz,1H),7.83(d,J=8.5Hz,1H),7.61–7.57(m,1H),7.33(dd,J=8.4,1.5Hz,1H),7.28(s,2H),7.23–7.18(m,1H),7.13(d,J=1.6Hz,1H),6.95(d,J=2.5Hz,1H),4.55–4.44(m,1H),3.95(d,J=5.2Hz,1H),3.81(t,J=11.3Hz,1H),3.61(s,3H),3.05(d,J=13.8,7.5Hz,1H),2.84(dd,J=13.8,5.6Hz,1H),2.82(d,J=5.6Hz,1H),1.42–1.37(m,1H),1.36–1.32(m,1H),0.96(s,3H),0.91(s,3H).
中间体29:(1R,5S)-N-((S)-1-羟基-3-苯丙-2-基)-6,6-二甲基-3-(喹啉-2-羰基)-3-氮杂双环[3.1.0]己烷-2-甲酰胺的制备
将中间体28(((1R,5S)-6,6-二甲基-3-(喹啉-2-羰基)-3-氮杂双环[3.1.0]己烷-2-羰基)-L-苯基丙氨酸)500mg,溶于30mL干燥甲醇中,常温条件下加入硼氢化钠,搅拌3小时后,加水淬灭,旋干甲醇,用乙酸乙酯萃取水相(50mL×3),收集有机相用硫酸钠进行干燥,抽滤后旋干有机相是为中间体29,产率80%。
化合物9:(1R,5S)-6,6-二甲基-N-((S)-1-氧-3-苯丙-2-基)-3-(喹啉-2-羰基)-3-氮杂双环[3.1.0]己烷-2-甲酰胺的制备
将中间体29(1R,5S)-N-((S)-1-羟基-3-苯丙-2-基)-6,6-二甲基-3-(喹啉-2-羰基)-3-氮杂双环[3.1.0]己烷-2-甲酰胺200mg,溶于10mL干燥二氯甲烷中,常温搅拌条件下加入戴斯马丁氧化剂,TLC监控反应进行完毕,抽滤除去氧化剂,滤液柱层析得化合物9,产率65%。1H NMR(400MHz,DMSO)δ9.55(s,1H),9.06(s,1H),8.56(d,J=7.3Hz,1H),8.44(dd,J=8.5,2.4Hz,1H),7.86(d,J=8.5Hz,1H),7.76(m,1H),7.32–7.21(m,5H),7.14(d,J=2.9Hz,,1H),7.08(d,J=5.7Hz,1H),5.33(s,1H),4.44(d,J=5.6Hz,1H),4.33(m,2H),4.13(m,2H),1.82(m,1H),1.50(m,1H),1.02(s,3H),0.87(s,3H)。
实施例4:制备化合物14
按照上述制备路线制得本发明的化合物14,路线中各步骤的反应条件如下:
i、三氟乙酸,二氯甲烷,25℃
ii、2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯,N,N-二异丙基乙胺,N,N-二甲基甲酰胺,室温。
iii、氢氧化钠,甲醇,水,55度
iv、2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯,N,N-二异丙基乙胺,N,N-二甲基甲酰胺,0℃
v、硼氢化钠,甲醇,室温
vi、戴斯马丁氧化剂,超干二氯甲烷,室温。
以下为具体合成步骤:
中间体30:甲基(S)-2-氨基-3-((S)-2-羰基-3-基)丙酸酯三氟乙酸盐的制备
将(S)-甲基2-(叔-丁氧基羰基氨基)-3-((S)-2-羰基吡咯烷-3-基)丙酸酯(中间体14,2.5g)溶于30mL二氯甲烷中,再加入20mL三氟乙酸,常温搅拌14小时,然后直接旋干得粗产品直接用于下一步反应。
中间体32:甲基(1R,2S,5S)-6,6-二甲基-3-(2-(4-(三氟甲氧基)苯氧)乙酰)-3-氮杂双环并[3.1.0]己烷-2-甲酸的制备
将2-(4-(三氟甲氧基)苯氧)乙酸(商业购得中间体31,0.24g,1.0mmol),2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(0.49g,1.2mmol)和N,N-二异丙基乙胺(494μL,3mmol)溶于一定N,N-二甲基甲酰胺中,然后加入(1R,2S,5S)-6,6-二甲基-3-氮杂双环[3.1.0]己烷-2-羧酸甲酯盐酸盐(商业购得中间体24,0.21g,1.0mmol).氩气保护常温反应12小时.反应加入4倍体积水,用二氯甲烷萃取三次,合并有机相用饱和氯化铵溶液、饱和碳酸钠溶液洗涤,无水硫酸钠干燥后过滤,硅胶拌样柱层析(石油醚/乙酸乙酯=1:1)得到白色固体中间体32(0.34g,88%)。1H NMR(400MHz,MeOD)δ7.19(d,J=8.9Hz,2H),7.00(d,J=8.7Hz,2H),4.80-4.71(m,2H),4.78–4.72(m,1H)3.89-3.72(m,1H),3.73(s,3H),3.67-3.60(m,1H),1.61-1.55(m,1H),1.49(d,J=7.4Hz,1H),1.08(s,3H),0.97(s,3H).ESI-MS(m/z):389.08(M+H)+.
中间体33:(1R,2S,5S)-6,6-二甲基-3-(2-(4-(三氟甲氧基)苯氧)乙酰)-3-氮杂双环并[3.1.0]己烷-2-甲酸的制备
用30mL甲醇溶解甲基(1R,2S,5S)-6,6-二甲基-3-(2-(4-(三氟甲氧基)苯氧)乙酰)-3-氮杂双环[3.1.0]己烷-2-甲酸(中间体32,200mg),然后加入2M NaOH溶液20mL.常温搅拌2.5小时.TLC监控反应结束后,旋干甲醇,用盐酸调pH至弱酸性,二氯甲烷萃取三次,合并有机相无水硫酸钠干燥后选干得粗产品直接进行下一步反应。
中间体34:甲基(1R,2S,5S)-6,6-二甲基-3-(2-(4-(三氟甲氧基)苯氧)乙酰)-3-氮杂双环并[3.1.0]己烷--2-酰胺)-3-((S)-2-羰基-3-基)丙酸酯的制备
在0℃反应条件下,往(1R,2S,5S)-6,6-二甲基-3-(2-(4-(三氟甲氧基)苯氧)乙酰)-3-氮杂双环[3.1.0]己烷-2-甲酸(中间体33,0.45g,1.2mmol)的超干DMF溶液中,加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(0.61g,1.6mmol),搅拌30分钟后,加入N.N-二异丙基乙胺(0.59mL,3.6mmol).然后,中间体14粗产品(0.27g1.45mmol)加入反应体系。反应在0℃氩气保护条件下搅拌12小时。TLC监控反应完后,加入4倍体积的水,用乙酸乙酯萃取三次,合并有机相用饱和氯化铵溶液、饱和碳酸氢钠溶液洗涤,无水硫酸钠干燥后过滤,拌样柱层析(乙酸乙酯/甲醇=10:1)得到白色固体是为中间体34.1H NMR(400MHz,MeOD)δ7.17(d,J=8.0Hz,2H),6.97(d,J=8.9Hz,2H),4.80–4.67(m,2H),4.55(d,J=11.8Hz,1H),3.94-3.83(m,1H),3.72(s,3H),3.68–3.57(m,1H),3.23–3.12(m,1H),3.11–3.00(m,2H),2.58(d,J=8.8Hz,1H),2.26–2.04(m,2H),1.73–1.40(m,4H),1.10(s,3H),0.92(s,3H).ESI-MS(m/z):542.13(M+H)+.
中间体35:(1R,2S,5S)-N-((S)-1-羟基-3-((S)-2-羰基-3-基)丙酸酯-2-基)-3-(2-(4-(三氟甲氧基)苯氧)乙酰基)-3-氮杂双环并[3.1.0]己烷-2-甲酰胺的制备
将甲基(1R,2S,5S)-6,6-二甲基-3-(2-(4-(三氟甲氧基)苯氧)乙酰)-3-氮杂双环[3.1.0]己烷--2-酰胺)-3-((S)-2-羰基-3-基)丙酸酯(中间体34 0.56mg,1.1mmol)加入50毫升中,低温条件下分批加入硼氢化钠(0.14g,8.8mmol).反应常温条件下搅拌2小时后,加水淬灭,旋干甲醇,用乙酸乙酯(50mL×3)对剩余水相进行萃取.有机相合并后用无水硫酸钠干燥,过滤旋干得白色固体为粗产品,被直接用于下一步反应。
化合物14:(1R,2S,5S)-6,6-二甲基-N-((S)-1-醛基-3-((S)-2-羰基-3-基)丙酸酯-2-基)-3-(2-(4-(三氟甲氧基)苯氧)乙酰基)-3-氮杂双环并[3.1.0]己烷-2-甲酰胺的制备
将(1R,2S,5S)-N-((S)-1-羟基-3-((S)-2-羰基-3-基)丙酸酯-2-基)-3-(2-(4-(三氟甲氧基)苯氧)乙酰基)-3-氮杂双环[3.1.0]己烷-2-甲酰胺(中间体36,(0.38g,0.75mmol)溶于超干二氯甲烷中,然后分批加入戴斯马丁氧化剂(0.95mg,0.79mmol),室温条件下反应3.5小时,TLC监控反应结束,反应体系过滤,用硫代硫酸钠溶液和饱和碳酸氢钠溶液洗涤有机相,浓缩后用制备色谱分离体系(乙腈/水=30:70)分离得化合物14(0.28g,45%)as a white solid.1H NMR(400MHz,MeOD)δ7.24–7.13(m,2H),7.04–6.92(m,2H),4.49(d,J=9.1Hz,1H),4.37-4.30(m,1H),4.06–3.89(m,1H),3.65-3.49(m,1H),3.11–2.98(m,1H),2.55(d,J=9.5Hz,1H),2.29–2.10(m,1H),2.06–1.99(m,1H),1.72–1.44(m,4H),1.13(s,3H),1.00(s,3H).13C NMR(101MHz,MeOD)δ181.60,172.58,167.18,156.90,142.99,121.99,115.47(d,J=7.7Hz),66.06,61.05,60.14,51.26,46.00,39.94,37.75,30.87(d,J=7.8Hz),29.88(d,J=18.5Hz),27.66(d,J=17.9Hz),25.03,19.04,11.63.HRMS(m/z):calculated for C24H28F3N3O6 +[M+H]+512.1964;found,512.2137.
实施例5:化合物15的制备
按照上述制备路线制得本发明的化合物15,路线中各步骤的反应条件如下:
i、2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯,N,N-二异丙基乙胺,N,N-二甲基甲酰胺,0℃
ii、硼氢化钠,甲醇,室温
iii、戴斯马丁氧化剂,超干二氯甲烷,室温。
具体合成步骤如下:
中间体36:甲基(S)-2-((1S,3aR,6aS)-2-(2-(2,4-二氯苯氧)乙酰基)八氢环戊烯并[c]吡咯-1-甲酰胺)-3-((S)-2-羰基-3-基)丙酸酯的制备
首先,2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(0.099g,0.26mmol)被加入(1S,3aR,6aS)-2-(2-(2,4-二氯苯氧)乙酰基)八氢环戊烯并[c]吡咯-1-羧酸(中间体23,0.071g,0.20mmol)的超干N,N-二甲基甲酰胺溶液中,反应体系先搅拌30分钟,N,N-二异丙基乙胺(100μL,0.60mmol),甲基(S)-2-氨基-3-((S)-2-羰基-3-基)丙酸酯三氟乙酸盐(中间体30,0.060g,0.32mmol)被依次加入反应体系.反应在0℃氩气保护条件下反应12小时,TLC监控反应结束,加入4倍体积的水,用乙酸乙酯萃取三次,合并有机相用饱和氯化铵溶液、饱和碳酸氢钠溶液洗涤,无水硫酸钠干燥后过滤,拌样柱层析(乙酸乙酯/甲醇=10:1)获得中间体36(0.063g,59%).1H NMR(400MHz,MeOD)δ7.40(d,J=2.5Hz,1H),7.26–7.19(m,1H),6.95(d,J=8.9Hz,1H),4.79-4.71(m,2H),4.54(d,J=3.7Hz,1H),4.25(t,J=4.3Hz,1H),3.72(s,3H),3.54–3.46(m,2H),3.21–3.12(m,1H),3.10–2.99(m,1H),2.86-2.70(m,2H),2.04–1.97(m,2H),1.96-1-85(m,3H),1.83–1.46(m,6H).ESI-MS(m/z):526.03(M+H)+.
中间体37:(1S,3aR,6aS)-2-(2-(2,4-二氯苯氧)乙酰基)-N-((S)-1-羟基-3-((S)-2-羰基-3-基)丙酸酯-2-基)八氢环戊烯并[c]吡咯-1-甲酰胺的制备
将甲基(S)-2-((1S,3aR,6aS)-2-(2-(2,4-二氯苯氧)乙酰基)八氢环戊烯并[c]吡咯-1-甲酰胺)-3-((S)-2-羰基-3-基)丙酸酯(中间体36,1,00g,2.0mmol)溶于无水甲醇中,然后再0℃条件下分批加入硼氢化钠(0.6g,16mmol),然后将温度升至室温,继续搅拌2小时,TLC检测反应结束,加水淬灭,旋干甲醇,用乙酸乙酯(50mL×3)对剩余水相进行萃取.有机相合并后用无水硫酸钠干燥,过滤旋干得白色固体为粗产品,被直接用于下一步反应。
化合物15:(1S,3aR,6aS)-2-(2-(2,4-二氯苯氧)乙酰基)-N-((S)-1-醛基-3-((S)-2-羰基-3-基)丙酸酯-2-基)八氢环戊烯并[c]吡咯-1-甲酰胺的制备
在(1S,3aR,6aS)-2-(2-(2,4-二氯苯氧)乙酰基)-N-((S)-1-羟基-3-((S)-2-羰基-3-基)丙酸酯-2-基)八氢环戊烯并[c]吡咯-1-甲酰胺(中间体37,0.50g,1.0mmol)的超干二氯甲烷溶液中,分批缓慢加入戴斯马丁氧化剂(0.55g,1.3mmol),室温条件下反应3.5小时,TLC监控反应结束,反应体系过滤,用硫代硫酸钠溶液和饱和碳酸氢钠溶液洗涤有机相,浓缩后用制备色谱分离体系(乙腈/水=45:55)得到白色固体化合物30(0.21g,42%)。1H NMR(400MHz,MeOD)δ7.42(t,J=4.3Hz,1H),7.28–7.17(m,1H),7.03–6.90(m,1H),4.47(dd,J=9.6,6.1Hz,1H),4.26(t,J=5.8Hz,1H),4.03–3.86(m,2H),3.51(dd,J=10.4,4.0Hz,1H),3.15(t,J=8.4Hz,1H),3.04–2.82(m,2H),2.75–2.50(m,2H),2.18(dd,J=13.2,7.0Hz,1H),2.18(dd,J=13.2,7.0Hz,1H),2.08–1.77(m,5H),1.76–1.43(m,5H).13CNMR(101MHz,MeOD)δ181.64,173.39,167.10(d,J=2.9Hz),152.81,129.30,127.35,125.73,123.12,114.81,66.81,60.14,53.90(d,J=35.6Hz),52.20,51.20(d,J=18.6Hz),43.34,40.00,37.73,31.69(d,J=4.2Hz),31.14(d,J=2.6Hz),29.62(d,J=24.1Hz),27.67,24.69,19.48,13.09.HRMS(m/z):calculated for C23H27Cl2N3O5 +[M+H]+496.1361;found,496.0842.
实施例6:化合物42的制备
按照上述制备路线制得本发明的化合物42,路线中各步骤的反应条件如下:
i、LDA,ClCH2I,THF
ii、HCl二氧六环溶液
iii、2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯,N,N-二异丙基乙胺,N,N-二甲基甲酰胺,0℃。
具体合成步骤如下:
中间体38:叔丁基((S)-4-氯-3-氧-1-((S)-2-羰基-3-基)丁烷-2-基)氨基甲酸酯的制备
选取干燥三口瓶,分别准备氩气保护和温度计,加入(S)-甲基2-(叔-丁氧基羰基氨基)-3-((S)-2-羰基吡咯烷-3-基)丙酸酯(中间体16,5g,17.5mmol),四氢呋喃(50mL),氯碘甲烷(5mL,68mmol)在-77℃下进行搅拌.二异丙基氨基锂(70mL,105mmol)被进一步滴入。加完以后进一步反应2小时,然后再低温条件下加入乙酸和四氢呋喃进行淬灭,产生的黑色悬浮物被进一步搅拌10分钟同时升温至室温。反应进一步用乙酸乙酯稀释,用水、饱和碳酸氢钠溶液和饱和食盐水洗涤,无水硫酸钠干燥后,过滤浓缩拌样柱层析得浅黄色固体为中间体38。1H NMR(400MHz,DMSO-d6)δ7.89(s,1H),7.72(d,J=7.5Hz,1H),4.72-4.94(m,2H),4.35(m,1H),3.26-3.40(m,2H),2.45(m,1H),2.32-2.42(m,1H),2.00-2.14(m,1H),1.79-1.99(m,2H),1.61(s,9H)。
中间体39:(S)-3-((S)-2-氨基-4-氯-3-氧丁基)吡咯烷-2-酮盐酸盐的制备
将叔丁基((S)-4-氯-3-氧-1-((S)-2-羰基-3-基)丁烷-2-基)氨基甲酸酯(中间体38)250mg加入20mL二氧六环中,随后加入20mL氯化氢二氧六环溶液20mL,常温搅拌4小时后,TLC检测反应完,旋干反应液得粗产品用于下一步反应。
化合物42:(1S,3aR,6aS)-N-((S)-4-氯-3-氧-1-((S)-2-羰基-3-基)丁烷-2-基)-2-(2-(2,4-二氯苯氧)乙酰基)八氢环戊烯并[c]吡咯-1-甲酰胺的制备
将2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(1g,2.6mmol)加入(1S,3aR,6aS)-2-(2-(2,4-二氯苯氧)乙酰基)八氢环戊烯并[c]吡咯-1-羧酸(中间体23,0.71g,2.0mmol)的超干N,N-二甲基甲酰胺溶液中,反应体系先搅拌30分钟,N,N-二异丙基乙胺(1mL,6.0mmol),(S)-3-((S)-2-氨基-4-氯-3-氧丁基)吡咯烷-2-酮盐酸盐(中间体39,0.652g,3.2mmol)被依次加入反应体系.反应在0℃氩气保护条件下反应16小时,TLC监控反应结束,加入3倍体积的水,用乙酸乙酯萃取三次,合并有机相用饱和氯化铵溶液、饱和碳酸氢钠溶液洗涤,无水硫酸钠干燥后过滤,制备纯化系统(乙腈/水=30:70)分离获得化合物42.1H NMR(400MHz,MeOD)δ7.40(s,1H),7.20(dd,J=8.7,2.4Hz,1H),7.05–6.91(m,1H),5.00–4.89(m,1H),4.79(d,J=16.1Hz,1H),4.69–4.35(m,2H),4.25(m,1H),3.99–3.85(m,1H),3.58–3.45(m,1H),3.25–3.03(m,1H),2.93–2.77(m,1H),2.76–2.50(m,2H),2.45–2.24(m,1H),2.18(d,J=10.9Hz,1H),2.11–1.47(m,9H),1.39–1.27(m,1H).ESI-MS(m/z):544.07(M+H)+.
实施例7:化合物50的制备
按照上述制备路线制得本发明的化合物50,路线中各步骤的反应条件如下:
i、叔丁基乙异腈,乙酸,超干二氯甲烷;
ii、1M氢氧化钠溶液,甲醇;
iii、戴斯马丁氧化剂、超干二氯甲烷。
具体合成步骤如下:
中间体40:(3S)-1-(叔丁氨基)-3-((1S,3aR,6aS)-2-(2-(2,4-二氯)乙酰基)八氢环戊烯并[c]吡咯-1-甲酰胺)-1-氧代-4-((S)-2-羰基-3-基)丁烷-2-基羧酸的制备
首先,化合物15(0.40mmol)被溶于超干二氯甲烷中,然后依次加入乙酸(0.028g,0.47mmol),叔丁基异腈(0.43mmol)。反应在常温条件下搅拌24小时,减压蒸馏柱层析(二氯甲烷/甲醇=15:1)得到中间体40。1H NMR(400MHz,DMSO-d6)δ7.87(d,J=12.1Hz,1H),7.46(s,1H),7.44(d,J=1.4Hz,1H),7.37(t,J=4.6Hz,1H),7.27(dd,J=7.5,1.5Hz,1H),7.11(d,J=7.5Hz,1H),5.09(d,J=7.1Hz,1H),4.82(s,2H),4.36–4.28(m,2H),3.64(ddd,J=59.5,12.4,7.0Hz,2H),3.22(td,J=7.1,4.6Hz,2H),2.67–2.44(m,3H),2.09(s,3H),1.99–1.51(m,10H),1.27(s,9H)。
中间体41:(1S,3aR,6aS)-N-((2S)-4-(叔丁氨基)-3-羟基-4-氧代-1-((S)-2-羰基-3-基)丁烷-2-基)-2-(2-(2,4-二氯)乙酰基)八氢环戊烯并[c]吡咯-1-甲酰胺的制备
1M的氢氧化钠溶液被加入(0.5mL)中间体40(0.164mmol)的甲醇溶液中,反应在常温条件下搅拌2小时,用1M盐酸调pH到中性.旋干反应液,残渣用二氯甲烷溶解,用水萃取后旋干反应液得粗产品41直接用于下一步反应。
化合物50:(1S,3aR,6aS)-N-((S)-4-(叔丁氨基)-3,4-氧代-1-((S)-2-羰基-3-基)丁烷-2-基)-2-(2-2,4-二氯)乙酰基)八氢环戊烯并[c]吡咯-1-甲酰胺的制备
在中间体41的超干二氯甲烷溶液中,分批缓慢加入戴斯马丁氧化剂,室温条件下反应4小时,TLC监控反应结束,反应体系过滤,用硫代硫酸钠溶液和饱和碳酸氢钠溶液洗涤有机相,浓缩后用制备色谱分离体系(乙腈/水=50:50)得到白色固体化合物50。1H NMR(400MHz,MEOD)δ7.42(d,J=1.4Hz,1H),7.26(dd,J=7.5,1.5Hz,1H),7.12(d,J=7.5Hz,1H),4.84(s,2H),4.67(dt,J=11.9,7.0Hz,1H),4.36(dd,J=7.0,0.7Hz,1H),3.74–3.57(m,2H),3.23–3.13(m,2H),2.70–2.46(m,3H),2.15–1.54(m,10H),1.43(s,9H).ESI-MS(m/z):595.07(M+H)+。
根据实施例1~7中的制备路线,改变原料,制得本发明表1所示的剩余化合物。
表1本发明化合物的结构和表征数据
以下通过实验例证明本发明化合物的药理效果。
实验例1:本发明化合物对Mpro酶活力抑制水平的测试
(1)实验方法
将重组SARS-CoV-2Mpro(最终浓度为750nM)与每种化合物的系列稀释液混合在25μL分析缓冲液(20mM Tris–HCl,pH 7.5,150mM NaCl,1mM EDTA,2mM DTT)中,并孵育10分钟。通过添加最终浓度为20μM的25μL荧光底物(MCA-AVLQ↓SGFR-Lys(Dnp)-Lys-NH2)来启动反应,用酶标仪测定320nm(激发)/405nm(发射)处的荧光信号。计算加入不同浓度化合物的反应的Vmax与加入DMSO的反应的Vmax,并用其生成IC50曲线。对于每种化合物,在9种浓度和3个独立重复下测量抗SARS-CoV-2Mpro的半抑制浓度(IC50)值。所有实验数据均采用GraphPadPrism软件进行分析。
(2)实验结果
表2化合物对SARS-COV-2Mpro的酶活抑制效果
从表2和图1、图2、图3可以看出,本发明的化合物能够有效抑制SARS-CoV-2Mpro的活性,可以用来制备SARS-CoV-2Mpro抑制剂,制备抗新型冠状病毒的药物,以及制备预防和/或治疗新型冠状病毒的药物。
实验例2:本发明的化合物对SARS-COV-2感染Vero E6细胞导致细胞死亡的抑制实验
(1)实验方法
通过检测化合物对SARS-COV-2感染Vero E6细胞导致的细胞死亡的抑制效果,初步评价化合物的抗病毒活性。具体实验方案是:将Vero E6细胞以细胞密度为2×104细胞/孔,100μL/孔接种在96孔板内,于37℃,5%CO2培养箱中培育过夜。次日,每孔同时加入100μL药物和100μL病毒稀释液(MOI=1),设置不含药物的阳性对照和不含病毒的阴性对照,37℃,5%CO2培养,72h后,通过CCK-8试剂盒检测细胞存活率,计算药物对病毒复制的抑制率和半数起效浓度(EC50)值,所有实验设置3个独立重复,所有实验数据均采用GraphPadPrism软件进行分析。
(2)实验结果
表3本发明化合物对SARS-COV-2感染Vero E6细胞导致细胞死亡的抑制活性
注:NT代表未测试细胞活性
从表3可以看出,本发明的化合物能够有效抑制SARS-COV-2感染Vero E6细胞导致的细胞死亡,说明本发明的化合物可以有效抑制SARS-COV-2病毒在细胞内的复制。
实验例3:化合物对Vero E6细胞的毒性实验
(1)实验方法
使用Vero E6细胞进行化合物的细胞毒性评估。具体实验方案是:将Vero E6细胞以细胞密度为2×104细胞/孔,100μL/孔接种在96孔板内,于37℃,5%CO2培养箱中培育过夜。次日,每孔加入200μL含药培养基,化合物以200μM为初始浓度,5倍梯度稀释,共6个梯度,每个浓度设置3个重复孔,每组实验设置不含药物的阴性对照和空白对照。药物处理72h后,使用CCK-8试剂盒检测细胞活力,计算化合物对Vero E6细胞的毒性和细胞半数毒性浓度(CC50)值。所有实验数据均采用GraphPad Prism软件进行分析。
(2)实验结果
表4本发明化合物对Vero E6细胞的毒性
从表4可以看出,本发明的化合物对Vero E6细胞的毒性很低。
实验例4:化合物对SARS-COV-2在人肺泡上皮细胞中复制的抑制实验
(1)实验方法
对于RT-qPCR方法,将人肺泡上皮细胞以8×105个细胞/孔的密度接种到48孔板(200μL/孔)中,并生长过夜。然后用病毒感染(MOI=0.01)和不同浓度的化合物处理细胞。在37℃下孵育1小时后,将含有病毒-药物混合物的培养基除去,并用含有化合物的新鲜培养基替换。继续孵育48小时后,收集细胞上清液提取病毒RNA,将其进行RT-qPCR定量分析并计算药物对病毒复制的抑制率和EC50值。EC50值是使用GraphPad Prism 8.0软件中的剂量反应模型计算的,实验设置2个独立重复。
(2)实验结果
测试结果如图4所示,化合物14、15、26、43、44和45均在人肺泡上皮细胞中表现出纳摩尔级的抑制SARS-COV-2复制的活性,优于已报道的活性最高的SARS-COV-2Mpro抑制剂11b(Dai et al.,2020,Science.368(6497):1331-1335)和GC376(Ma et al.,2020,CellRes.30(8):678-692)在同样测试条件下的抗病毒活性(11b,EC50=23.6nM;GC376,EC50=151.3nM)。
实验例5:噬斑法评价化合物抗SARS-COV-2活性(Vero E6细胞)
(1)实验方法
使用噬斑法评价化合物3和39在Vero E6细胞中的抗SARS-COV-2活性。Vero E6以每孔1.0×105个接种于24孔细胞培养板中,37℃培养过夜后备用。加入梯度稀释的药物后,加入SARS-CoV-2感染细胞,MOI约为0.002。置于37℃细胞培养箱培养1小时后,去掉含药感染上清,PBS洗一遍,加入含有不同浓度药物的羧甲基纤维素钠0.5mL,羧甲基纤维素钠终浓度为0.9%,置于37℃细胞培养箱培养72小时。用20%的甲醛固定2小时,加入0.5%的结晶紫染色20分钟后,晾干拍照,观察空斑大小并记录空斑数目。实验设空白对照孔(正常细胞),病毒对照孔,阳性药物对照孔。
计算公式:抑制率(%)=(病毒对照孔空斑数-样品孔空斑数)/病毒对照孔空斑数*100
计算所得的细胞活性和抑制率,用Graphpad Prism8计算EC50(半数起效浓度)值。
(2)实验结果
表5小分子化合物对SARS-CoV-2的抑制作用
化合物名称 | <![CDATA[EC<sub>50</sub>(μM)]]> |
3 | 0.24 |
39 | 1.20 |
Remdesivir(瑞德西韦) | 0.69 |
实验结果如表5所示,本发明化合物可以有效抑制Vero E6细胞中SARS-COV-2感染;特别是化合物3,EC50为0.2373μM,活性优于阳性对照瑞德西韦(EC50为0.692μM)。
实验例6:化合物对大鼠的体内药代动力学特性评估
(1)给药方案
雄性Sprague-Dawley(SD)大鼠60只,体重200-230g,随机分成3组,每组3只。按照如下表6方案分别灌胃(p.o.)、静脉(i.v.)和腹腔(i.p.)给予系列受试化合物。实验前禁食12h,自由饮水。给药后2h统一进食。
灌胃、静脉和腹腔给药溶液以DMSO/HS15/NaCl(5/3/92,v/v/v)配制。按表6所示给药剂量给予药物,记录给药时间,并在以上设定的时间点经颈静脉采血或其他合适方式,每个样品采集约0.20mL,肝素钠抗凝,采集后放置冰上。并于1小时之内离心分离血浆(离心条件:6800g,6分钟,2-8℃)。血浆样本在分析前存放时则放于-80℃冰箱内。分组及采血时间点见表6,每个时间点3只动物。
表6化合物对大鼠的体内药代动力学特性评估实验方案
(2)实验结果
表7化合物的主要药动学参数
结果如表7所示。本发明对化合物3,14,15,26,39,40,43,44和45进行了药代动力学研究。其中,化合物3的口服暴露量为2293h*ng/mL,生物利用度为55.1%。化合物14的腹腔暴露量为11581h*ng/mL,生物利用度为78.0%;口服暴露量为1665h*ng/mL,生物利用度为11.2%。化合物15的腹腔注射给药暴露量为12166h*ng/mL,生物利用度为62.3%;口服暴露量为2843h*ng/mL,生物利用度为14.6%。化合物26的口服暴露量为842h*ng/mL,生物利用度为7.2%。化合物39的口服暴露量为14586h*ng/mL,生物利用度为14.7%。化合物40的口服暴露量为2888h*ng/mL,生物利用度为22.1%。化合物43的口服暴露量为258h*ng/mL,生物利用度为4.8%。化合物44的口服暴露量为381h*ng/mL,生物利用度为4.1%。化合物45的口服暴露量为968h*ng/mL,生物利用度为5.1%。
实验结果表明,本发明化合物在大鼠的体内具有良好的药代动力学性质。
实验例7:化合物对大鼠的体内安全性的初步评价
(1)实验方法
将化合物溶解在5%(v/v)DMSO(Sigma-Aldrich),3%(v/v)HS15(GLPBIO)和92%生理盐水中。SPF SD大鼠(年龄:7-11周)雌190-220克)雄(体重200-230克)各半。按表8的给药方案进行试验,并对所有动物进行临床观察。并在实验结束时,收集心脏,肝,脾,肺,肾和给药部位的样品。试验结果如表8所示。
(2)实验结果
表8对大鼠的体内安全性的初步评价
实验结果显示,本发明化合物对大鼠的体内安全性良好。
实验例8:化合物对转基因小鼠的体内抗SARS-COV-2感染的活性研究
(1)实验方案
人源化血管紧张素转化酶2(ACE2)转基因小鼠(年龄:8-10周)购自江苏集萃药康生物科技有限公司(#T037659。将化合物溶于5%(v/v)DMSO(Sigma-Aldrich),3%(v/v)HS15(GLPBIO)和92%生理盐水中。按表9的试验方案进行SARS-CoV-2(stain107)滴鼻感染和给药。观察所有小鼠并每天监测其体重直至处死。病毒感染后第1(1dpi)、3(3dpi)和5(5dpi)天,收集肺组织(n=3,每个dpi组)用于病毒载量检测、H&E组织病理学分析、代表性炎性细胞因子和趋化因子测定及炎性细胞(中性粒细胞和巨噬细胞)计数。
表9化合物对转基因小鼠体内抗SARS-COV-2感染的活性研究方案
肺部病毒载量检测的具体实验方案是:使用TRIzolTM试剂(Invitrogen)从肺组织中提取RNA,并使用一步法qRT-PCR试剂盒(Toyobo)对病毒RNA进行定量,结果表示为每微克组织的病毒RNA的拷贝数。
肺部组织病理学分析的具体实验方案是:将肺部组织用4%多聚甲醛固定至少7天,包埋在石蜡中并切成3μm的切片。将切片用苏木精和曙红(H&E)染色,并通过光学显微镜进行分析。根据组织学特征(肺泡间隔增厚,出血,炎性细胞浸润等)评估肺损伤。
肺部代表性炎性细胞因子和趋化因子测定的具体实施方案是:使用PrimeScriptTMRT试剂盒(Takara),将从肺部提取的RNA反转录为cDNA,然后通过Ex TaqTMII(TliRNaseH Plus)(Takara)和ViiATM定量基因表达。表10中显示了用于定量炎性基因表达的引物序列。
表10测定代表性炎性细胞因子和趋化因子的引物序列
肺部测定炎性细胞(中性粒细胞和巨噬细胞)数量的具体实施方案是:将小鼠肺部组织在4%多聚甲醛中固定至少7天,然后按照标准程序将石蜡包埋并切成4μm切片。在二甲苯中进行去石蜡,抗原回收和封闭后,将肺切片与大鼠单克隆抗体F4/80(Huabio,1:100)或兔多克隆抗体Ly6G(Servicebio,1:300)在4℃孵育过夜,然后与辣根过氧化物酶(HRP)偶联的山羊抗大鼠二抗或HRP偶联的山羊抗兔二抗在室温下反应1小时,以根据酪酰胺信号放大(TSA)催化Cy3-酪胺和Cy5-酪胺并放大染色信号。用DAPI对细胞核染色后,所有切片均使用LEICA DMI 4000B显微镜(德国)拍照,并通过ImageJ软件(美国NIH)和FlowJo软件(美国BD)进行分析。为了半定量地测量巨噬细胞和嗜中性白细胞的浸润,通过光学显微镜检查每个肺切片中5个任意选择的肺实质区域,以观察嗜中性白细胞或巨噬细胞的存在。该评估以盲法进行。每只动物的累积得分表示为每100个视野的阳性视野数(%)。
上述实验以安慰剂作为对照。该安慰剂为与受试药物剂型相同、但是不含药物活性成分的制剂。
(2)实验结果
肺部病毒载量检测实验结果如图5所示,口服和腹腔给予化合物14及腹腔给予化合物15均可以有效降低SARS-COV-2感染的转基因小鼠肺部的病毒载量。
肺部组织病理学分析实验结果如图6所示,口服和腹腔给予化合物14及腹腔给予化合物15均可以有效改善SARS-COV-2感染的转基因小鼠肺部的病理损伤。
肺部代表性炎性细胞因子和趋化因子测定实验结果如图7所示,口服和腹腔给予化合物14及腹腔给予化合物15可以有效降低肺部趋化因子配体10(CXCL10)和β型干扰素(IFN-β)的基因表达水平。
肺部测定炎性细胞(中性粒细胞和巨噬细胞)数量的实验结果如图8所示,口服和腹腔给予化合物14及腹腔给予化合物15均可以有效降低SARS-COV-2感染的转基因小鼠肺部的中性粒细胞(NEU)和巨噬细胞(MAC)的数量。
实验结果表明,本发明化合物能够有效抵抗转基因小鼠的体内SARS-COV-2感染。
综上,本发明提供了一种式I所示的新型冠状病毒主蛋白酶抑制剂及其制备方法和用途。式I所示化合物能够有效抑制SARS-CoV-2Mpro活性,可以用来制备SARS-CoV-2Mpro抑制剂,阻断SARS-CoV-2病毒在患者体内的复制和转录。本发明的化合物在制备SARS-CoV-2Mpro抑制剂,抗SARS-CoV-2的药物,以及预防和/或治疗新型冠状病毒的药物中具有非常好的应用前景。
Claims (9)
1.式I所示化合物、或其药学上可接受的盐、或其立体异构体、或其旋光异构体、或其同位素替代形式:
其中,X为O或S;
A环选自未取代或被一个或多个R6取代的以下基团:5~6元饱和杂环基、5~6元不饱和杂环基、饱和杂稠环基、不饱和杂稠环基;R6各自独立的选自C1~6烷基、C1~6烷氧基、卤素、羟基、氰基、氨基、羧基;
R3为L3M0L4R3a;其中L3选自无、C1~4亚烷基、卤代C1~4亚烷基、C2~4亚烯基、卤代C2~4亚烯基,L4选自无、C1~4亚烷基、卤代C1~4亚烷基,M0选自无、O、S、NH、CO、CONH、NHCO,R3a为未取代或被一个或多个R3b取代的以下基团:5~6元芳基、5~6元杂芳基、不饱和杂稠环基、不饱和稠环烷基;R3b各自独立的选自被R3c取代或未取代的C1~5烷基、被R3c取代或未取代的C1~5烷氧基、卤素、被R3c取代或未取代的苯基、NR14R15、被R3c取代或未取代的萘基、羟基;R14、R15各自独立的选自氢或C1~5烷基,R3c各自独立的选自卤素、氘、氰基、羟基、氨基、羧基;
R4选自未取代或被一个或多个取代基取代的以下基团:5~6元芳基、5~6元杂芳基、C1~5烷基、COOR10;所述取代基各自独立的选自=O、羟基、硝基、氨基、羧基、卤素、C1~5烷基;R10为C1~5烷基;
R5选自COR8或WCOOR7;其中,R8选自氢或W选自无、C1~4亚烷基、C2~4亚烯基、C2~4亚炔基,R7选自C1~6烷基;M选自无、CO、NH、CONH、NHCO、COO或OCO,L0选自无、C1~4亚烷基、C2~4亚烯基,L1选自无、C1~4亚烷基、C2~4亚烯基,R8a选自C1~5烷基、卤代的C1~5烷基、3~6元饱和环烷基、3~6元饱和杂环基、5~6元芳基或5~6元杂芳基。
2.根据权利要求1所述的化合物、或其药学上可接受的盐、或其立体异构体、或其旋光异构体、或其同位素替代形式,其特征在于:所述化合物的结构如式II、式III或式IV所示:
其中,X为O或S;
n选自0~3的整数,优选为0~2的整数;
R1、R2各自独立的选自氢、C1~5烷基、C1~5烷氧基、卤素、羟基、氰基、氨基、羧基;
R3为L3M0L4R3a;其中L3选自无、C1~4亚烷基、卤代C1~4亚烷基、C2~3亚烯基,L4选自无、C1~4亚烷基、卤代C1~4亚烷基,M0选自无、O、S、NH、CO、CONH、NHCO,R3a为未取代或被一个或多个R3b取代的以下基团:苯基、
R3b各自独立的选自C1~4烷基、卤素取代的C1~4烷基、氘代的C1~4烷基、氰基取代的C1~4烷基、C1~4烷氧基、卤素取代的C1~4烷氧基、氘代的C1~4烷氧基、氰基取代的C1~4烷氧基、卤素、苯基、卤代的苯基、NR14R15、羟基,R14、R15各自独立的选自氢或C1~4烷基;
R4选自未取代或被一个或多个取代基取代的以下基团:5~6元芳基、5~6元杂芳基、C1~5烷基、COOR10;所述取代基各自独立的选自=O、羟基、硝基、氨基、羧基、卤素、C1~5烷基;R10为C1~5烷基;
R8选自氢或M选自无、CO、NH、CONH、NHCO、COO或OCO,L0选自无、C1~3亚烷基、C2~4亚烯基,L1选自无、C1~3亚烷基、C2~4亚烯基,R8a选自C1~4烷基、卤代的C1~4烷基、3~6元饱和环烷基、3~6元饱和杂环基、5~6元芳基或5~6元杂芳基。
3.根据权利要求2所述的化合物、或其药学上可接受的盐、或其立体异构体、或其旋光异构体、或其同位素替代形式,其特征在于:
R1、R2各自独立的选自氢、C1~4烷基、C1~4烷氧基、卤素、羟基;
R3选自 L3M0L4R3a;L3选自无、C1~3亚烷基、卤代C1~3亚烷基、C2~3亚烯基,L4选自无、C1~3亚烷基、卤代C1~3亚烷基、,M0选自无、O、NH、CO、CONH,R3a为苯基、被一个或多个R3b取代的苯基,R3b各自独立的选自C1~4烷基、卤素取代的C1~4烷基、氘代的C1~4烷基、氰基取代的C1~4烷基、C1~4烷氧基、卤素取代的C1~4烷氧基、氘代的C1~4烷氧基、氰基取代的C1~4烷氧基、卤素、苯基、卤代的苯基、NR14R15、羟基,R14、R15各自独立的选自氢或C1~3烷基;
R4选自C1~2烷基、COOR10、取代或未取代的苯基;所述取代基选自羟基、硝基;Ra1、Ra2各自独立的选自氢、C1~3烷基、卤素;R10为C1~3烷基;
R8选自氢、CONHR11、L2COOR12、C1~4烷基、卤代的C1~4烷基;R11选自3~6元饱和环烷基、C1~4烷基、苄基、L2为C1~2亚烷基、C2~3亚烯基,R12为C1~3烷基。
4.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、或其立体异构体、或其旋光异构体、或其同位素替代形式,其特征在于:所述化合物的结构为以下结构之一:
5.一种药物组合物,其特征在于:所述药物组合物是以权利要求1~4任一项所述化合物、或其药学上可接受的盐、或其立体异构体、或其旋光异构体、或其同位素替代形式为活性成分,加上药学上可接受的辅料制成的制剂。
6.权利要求1~4任一项所述化合物、或其药学上可接受的盐、或其立体异构体、或其旋光异构体、或其同位素替代形式在制备冠状病毒蛋白水解酶抑制剂中的用途;优选的,所述冠状病毒蛋白水解酶为冠状病毒主蛋白酶;更优选的,所述冠状病毒蛋白水解酶为SARS-COV-2Mpro。
7.权利要求1~4任一项所述化合物、或其药学上可接受的盐、或其立体异构体、或其旋光异构体、或其同位素替代形式在制备抗冠状病毒的药物中的用途,优选的,所述冠状病毒为新型冠状病毒SARS-CoV-2。
8.权利要求1~4任一项所述化合物、或其药学上可接受的盐、或其立体异构体、或其旋光异构体、或其同位素替代形式在制备预防和/或治疗与SARS-COV-2Mpro相关的疾病的药物中的用途,优选的,所述与SARS-COV-2Mpro相关的疾病为新型冠状病毒COVID-19。
9.根据权利要求6~8任一项所述的用途,其特征在于:所述冠状病毒蛋白水解酶抑制剂、抗冠状病毒的药物或预防和/或治疗病毒性肺炎的药物能够抑制SARS-COV-2Mpro的活性和/或能够抑制SARS-COV-2感染细胞。
Applications Claiming Priority (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010761302 | 2020-07-31 | ||
CN202010761283 | 2020-07-31 | ||
CN2020107612837 | 2020-07-31 | ||
CN2020107613026 | 2020-07-31 | ||
CN2020109063980 | 2020-09-01 | ||
CN202010906398 | 2020-09-01 | ||
CN202011568282.7A CN114057702B (zh) | 2020-07-31 | 2020-12-25 | 一种新型冠状病毒主蛋白酶的抑制剂及其制备方法和用途 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202011568282.7A Division CN114057702B (zh) | 2020-07-31 | 2020-12-25 | 一种新型冠状病毒主蛋白酶的抑制剂及其制备方法和用途 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN115960088A true CN115960088A (zh) | 2023-04-14 |
Family
ID=80037080
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210944223.8A Pending CN115960088A (zh) | 2020-07-31 | 2020-12-25 | 一种新型冠状病毒主蛋白酶的抑制剂及其制备方法和用途 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN115960088A (zh) |
WO (1) | WO2022021841A1 (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115894504A (zh) * | 2022-10-21 | 2023-04-04 | 深圳信立泰药业股份有限公司 | 一种冠状病毒3cl蛋白酶抑制剂及其用途 |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU2021313130A1 (en) * | 2020-07-20 | 2023-02-09 | Enanta Pharmaceuticals, Inc. | Functionalized peptides as antiviral agents |
KR20230107825A (ko) * | 2021-04-16 | 2023-07-18 | 푸지엔 에이키링크 바이오테크놀로지 컴퍼니 리미티드 | 고리 변형된 프롤린 쇼트 펩티드 화합물 및 이의 용도 |
WO2022266363A1 (en) * | 2021-06-16 | 2022-12-22 | The Scripps Research Institute | Protease inhibitors for the treatment of coronavirus infections |
WO2023165334A1 (zh) * | 2022-03-01 | 2023-09-07 | 成都威斯克生物医药有限公司 | 酮酰胺类衍生物及其制药用途 |
CN116768967A (zh) * | 2022-03-07 | 2023-09-19 | 广州谷森制药有限公司 | 氘代内酰胺类化合物及其制备方法、组合物和用途 |
CN114957383A (zh) * | 2022-04-01 | 2022-08-30 | 中国科学院上海药物研究所 | 一种拟肽类化合物及其制备方法、药物组合物和用途 |
CN115490681B (zh) * | 2022-07-08 | 2023-04-18 | 歌礼生物科技(杭州)有限公司 | 三嗪衍生物 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105837487A (zh) * | 2016-03-17 | 2016-08-10 | 天津国际生物医药联合研究院 | MERS-CoV主蛋白酶小分子抑制剂及其制备方法和用途 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
SV2003000617A (es) * | 2000-08-31 | 2003-01-13 | Lilly Co Eli | Inhibidores de la proteasa peptidomimetica ref. x-14912m |
-
2020
- 2020-12-25 CN CN202210944223.8A patent/CN115960088A/zh active Pending
-
2021
- 2021-02-09 WO PCT/CN2021/076400 patent/WO2022021841A1/zh active Application Filing
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105837487A (zh) * | 2016-03-17 | 2016-08-10 | 天津国际生物医药联合研究院 | MERS-CoV主蛋白酶小分子抑制剂及其制备方法和用途 |
Non-Patent Citations (2)
Title |
---|
HASAN M.: "Main protease inhibitors and drug surface hotspot for the treatment of COVID-19: drug repurposing and molecular docking approach", CHEMRXIV, 27 April 2020 (2020-04-27), pages 1 - 42 * |
TA GUDASHEVA: "Synthesis and antiamnesic activity of a series of N-acylprolyl-containing dipeptides", EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY, vol. 31, 31 December 1996 (1996-12-31), pages 151 - 157, XP004040102, DOI: 10.1016/0223-5234(96)80448-X * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115894504A (zh) * | 2022-10-21 | 2023-04-04 | 深圳信立泰药业股份有限公司 | 一种冠状病毒3cl蛋白酶抑制剂及其用途 |
Also Published As
Publication number | Publication date |
---|---|
WO2022021841A1 (zh) | 2022-02-03 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN114057702B (zh) | 一种新型冠状病毒主蛋白酶的抑制剂及其制备方法和用途 | |
CN115960088A (zh) | 一种新型冠状病毒主蛋白酶的抑制剂及其制备方法和用途 | |
KR102481876B1 (ko) | 니트릴-함유 항바이러스 화합물 | |
WO2022013684A1 (en) | Antiviral heteroaryl ketone derivatives | |
WO2016210247A1 (en) | New methods of use for an anti-diarrhea agent | |
CN108676009B (zh) | 作为her2酪氨酸激酶抑制剂的嘧啶衍生物及其应用 | |
WO2023165334A1 (zh) | 酮酰胺类衍生物及其制药用途 | |
CN115108970B (zh) | 二酰胺类衍生物及其制药用途 | |
RU2786722C1 (ru) | Нитрилсодержащие противовирусные соединения | |
WO2017076187A1 (zh) | 1,4(1,4)-二苯杂环六蕃-12,43-二基衍生物及其制备方法与应用 | |
TW202304888A (zh) | 醚連接之抗病毒化合物 | |
NZ791608A (en) | Nitrile-containing antiviral compounds | |
CN117836272A (zh) | 用于治疗或预防冠状病毒感染的3cl蛋白酶小分子抑制剂及其用途 | |
EA046321B1 (ru) | Нитрилсодержащие противовирусные соединения | |
JPWO2007136085A1 (ja) | スピロ四環系化合物 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |