CN100363346C - Small molecular inhibiting agent for coronavirus main proteinase, its preparation method and uses - Google Patents
Small molecular inhibiting agent for coronavirus main proteinase, its preparation method and uses Download PDFInfo
- Publication number
- CN100363346C CN100363346C CNB2004100877217A CN200410087721A CN100363346C CN 100363346 C CN100363346 C CN 100363346C CN B2004100877217 A CNB2004100877217 A CN B2004100877217A CN 200410087721 A CN200410087721 A CN 200410087721A CN 100363346 C CN100363346 C CN 100363346C
- Authority
- CN
- China
- Prior art keywords
- compound
- group
- formula
- coronavirus
- benzyl
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 230000002401 inhibitory effect Effects 0.000 title description 23
- 108700003471 Coronavirus 3C Proteases Proteins 0.000 title 1
- 241000711573 Coronaviridae Species 0.000 claims abstract description 20
- 239000003814 drug Substances 0.000 claims abstract description 8
- 150000001875 compounds Chemical class 0.000 claims description 66
- -1 t-butoxycarbonyl Chemical group 0.000 claims description 40
- 241000315672 SARS coronavirus Species 0.000 claims description 27
- WYURNTSHIVDZCO-UHFFFAOYSA-N tetrahydrofuran Substances C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 20
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 19
- 239000002904 solvent Substances 0.000 claims description 16
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 13
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-dimethylformamide Substances CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 13
- 241000711484 Transmissible gastroenteritis virus Species 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 13
- HEDRZPFGACZZDS-UHFFFAOYSA-N CHCl3 Substances ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 12
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 12
- 241000711475 Feline infectious peritonitis virus Species 0.000 claims description 12
- 241000711450 Infectious bronchitis virus Species 0.000 claims description 11
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 claims description 11
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 10
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 10
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 9
- 239000007821 HATU Substances 0.000 claims description 9
- 125000001584 benzyloxycarbonyl group Chemical group C(=O)(OCC1=CC=CC=C1)* 0.000 claims description 9
- 125000004175 fluorobenzyl group Chemical group 0.000 claims description 9
- UYWQUFXKFGHYNT-UHFFFAOYSA-N phenylmethyl ester of formic acid Natural products O=COCC1=CC=CC=C1 UYWQUFXKFGHYNT-UHFFFAOYSA-N 0.000 claims description 9
- 239000003795 chemical substances by application Substances 0.000 claims description 8
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 7
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 7
- 125000000590 4-methylphenyl group Chemical group [H]C1=C([H])C(=C([H])C([H])=C1*)C([H])([H])[H] 0.000 claims description 6
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 claims description 6
- 208000001528 Coronaviridae Infections Diseases 0.000 claims description 5
- 239000002253 acid Substances 0.000 claims description 5
- 239000000010 aprotic solvent Substances 0.000 claims description 5
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 5
- 150000007530 organic bases Chemical class 0.000 claims description 5
- 239000003960 organic solvent Substances 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 5
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- 125000001207 fluorophenyl group Chemical group 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 4
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 4
- 125000006239 protecting group Chemical group 0.000 claims description 4
- 125000004044 trifluoroacetyl group Chemical group FC(C(=O)*)(F)F 0.000 claims description 4
- 150000001732 carboxylic acid derivatives Chemical class 0.000 claims description 3
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 claims description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 3
- 238000011282 treatment Methods 0.000 claims description 3
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 claims description 2
- 125000004176 4-fluorobenzyl group Chemical group [H]C1=C([H])C(=C([H])C([H])=C1F)C([H])([H])* 0.000 claims description 2
- 238000009833 condensation Methods 0.000 claims description 2
- 230000005494 condensation Effects 0.000 claims description 2
- 238000001212 derivatisation Methods 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims description 2
- 241000282898 Sus scrofa Species 0.000 claims 1
- 125000005931 tert-butyloxycarbonyl group Chemical group [H]C([H])([H])C(OC(*)=O)(C([H])([H])[H])C([H])([H])[H] 0.000 claims 1
- 241000007181 unidentified human coronavirus Species 0.000 claims 1
- 239000003112 inhibitor Substances 0.000 abstract description 16
- 239000013078 crystal Substances 0.000 abstract description 9
- 208000015181 infectious disease Diseases 0.000 abstract description 3
- 108030001409 SARS coronavirus main proteinases Proteins 0.000 abstract 1
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 31
- 101800000535 3C-like proteinase Proteins 0.000 description 19
- 101800002396 3C-like proteinase nsp5 Proteins 0.000 description 19
- 150000003384 small molecules Chemical class 0.000 description 15
- 108091005804 Peptidases Proteins 0.000 description 12
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 12
- 239000000758 substrate Substances 0.000 description 11
- 239000004365 Protease Substances 0.000 description 9
- 238000006243 chemical reaction Methods 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 8
- 239000001257 hydrogen Substances 0.000 description 8
- 229910052739 hydrogen Inorganic materials 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- 229920006395 saturated elastomer Polymers 0.000 description 8
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 8
- 238000003818 flash chromatography Methods 0.000 description 7
- 238000001228 spectrum Methods 0.000 description 7
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 6
- 201000003176 Severe Acute Respiratory Syndrome Diseases 0.000 description 6
- 238000013480 data collection Methods 0.000 description 6
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 6
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 5
- 244000309467 Human Coronavirus Species 0.000 description 5
- 108010055591 SARS coronavirus 3C-like protease Proteins 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 238000001704 evaporation Methods 0.000 description 5
- 238000001914 filtration Methods 0.000 description 5
- 230000002209 hydrophobic effect Effects 0.000 description 5
- 239000000178 monomer Substances 0.000 description 5
- 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 4
- 239000007832 Na2SO4 Substances 0.000 description 4
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 4
- 102000035195 Peptidases Human genes 0.000 description 4
- 125000000217 alkyl group Chemical group 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 238000001035 drying Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 4
- 230000005284 excitation Effects 0.000 description 4
- 150000003951 lactams Chemical group 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 229910052938 sodium sulfate Inorganic materials 0.000 description 4
- QFLWZFQWSBQYPS-AWRAUJHKSA-N (3S)-3-[[(2S)-2-[[(2S)-2-[5-[(3aS,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]-3-methylbutanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-4-[1-bis(4-chlorophenoxy)phosphorylbutylamino]-4-oxobutanoic acid Chemical compound CCCC(NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)[C@@H](NC(=O)CCCCC1SC[C@@H]2NC(=O)N[C@H]12)C(C)C)P(=O)(Oc1ccc(Cl)cc1)Oc1ccc(Cl)cc1 QFLWZFQWSBQYPS-AWRAUJHKSA-N 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 241000282414 Homo sapiens Species 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 101710153041 Replicase polyprotein 1a Proteins 0.000 description 3
- 101710151619 Replicase polyprotein 1ab Proteins 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 229940125773 compound 10 Drugs 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000007062 hydrolysis Effects 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- ZLVXBBHTMQJRSX-VMGNSXQWSA-N jdtic Chemical compound C1([C@]2(C)CCN(C[C@@H]2C)C[C@H](C(C)C)NC(=O)[C@@H]2NCC3=CC(O)=CC=C3C2)=CC=CC(O)=C1 ZLVXBBHTMQJRSX-VMGNSXQWSA-N 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 3
- 238000012916 structural analysis Methods 0.000 description 3
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000711466 Murine hepatitis virus Species 0.000 description 2
- 101710200092 Replicase polyprotein Proteins 0.000 description 2
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 125000004432 carbon atom Chemical group C* 0.000 description 2
- 125000004185 ester group Chemical group 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 2
- 125000004108 n-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 2
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000003756 stirring Methods 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
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- DFPYXQYWILNVAU-UHFFFAOYSA-N 1-hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1.C1=CC=C2N(O)N=NC2=C1 DFPYXQYWILNVAU-UHFFFAOYSA-N 0.000 description 1
- KTPDNYVWXJTUKG-UHFFFAOYSA-O 1-methyl-2-[2-(2-phenyl-1h-indol-3-yl)ethenyl]quinolin-1-ium Chemical compound C1=CC2=CC=CC=C2[N+](C)=C1\C=C\C(C1=CC=CC=C1N1)=C1C1=CC=CC=C1 KTPDNYVWXJTUKG-UHFFFAOYSA-O 0.000 description 1
- QWENRTYMTSOGBR-UHFFFAOYSA-N 1H-1,2,3-Triazole Chemical compound C=1C=NNN=1 QWENRTYMTSOGBR-UHFFFAOYSA-N 0.000 description 1
- 125000004847 2-fluorobenzyl group Chemical group [H]C1=C([H])C(F)=C(C([H])=C1[H])C([H])([H])* 0.000 description 1
- 125000004198 2-fluorophenyl group Chemical group [H]C1=C([H])C(F)=C(*)C([H])=C1[H] 0.000 description 1
- 125000006284 3-fluorobenzyl group Chemical group [H]C1=C([H])C(=C([H])C(F)=C1[H])C([H])([H])* 0.000 description 1
- 125000004180 3-fluorophenyl group Chemical group [H]C1=C([H])C(*)=C([H])C(F)=C1[H] 0.000 description 1
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 1
- 125000001255 4-fluorophenyl group Chemical group [H]C1=C([H])C(*)=C([H])C([H])=C1F 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 241000711443 Bovine coronavirus Species 0.000 description 1
- 101100338269 Caenorhabditis elegans his-41 gene Proteins 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000711467 Human coronavirus 229E Species 0.000 description 1
- 241001292005 Nidovirales Species 0.000 description 1
- 239000008118 PEG 6000 Substances 0.000 description 1
- 229920002565 Polyethylene Glycol 400 Polymers 0.000 description 1
- 229920002584 Polyethylene Glycol 6000 Polymers 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000002528 anti-freeze Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 238000002447 crystallographic data Methods 0.000 description 1
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 description 1
- 125000002147 dimethylamino group Chemical group [H]C([H])([H])N(*)C([H])([H])[H] 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 150000002148 esters Chemical group 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 125000000623 heterocyclic group Chemical group 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 125000001280 n-hexyl group Chemical group C(CCCCC)* 0.000 description 1
- 125000000740 n-pentyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- JLFNLZLINWHATN-UHFFFAOYSA-N pentaethylene glycol Chemical compound OCCOCCOCCOCCOCCO JLFNLZLINWHATN-UHFFFAOYSA-N 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 230000000405 serological effect Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 230000029812 viral genome replication Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
- C07K5/0205—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -NH-(X)3-C(=0)-, e.g. statine or derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- 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
-
- 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
- C07D207/18—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 one double bond between ring members or between a ring member and a non-ring member
- C07D207/22—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 one double bond between ring members or between a ring member and a non-ring member 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
- C07D207/24—Oxygen or sulfur atoms
- C07D207/26—2-Pyrrolidones
- C07D207/263—2-Pyrrolidones with only hydrogen atoms or radicals containing only hydrogen and carbon atoms directly attached to other ring carbon atoms
- C07D207/27—2-Pyrrolidones with only hydrogen atoms or radicals containing only hydrogen and carbon atoms directly attached to other ring carbon atoms with substituted hydrocarbon radicals directly attached to the ring nitrogen atom
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
- C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Virology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pulmonology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention provides a series of small molecular inhibitors designed on the basis of crystal texture of SARS coronavirus main proteinase, and a texture general formula is disclosed as a formula (I). The present invention also provides a preparation method for the small molecular inhibitors and an application for preparing medicaments for treating or preventing the infection of various kinds of coronavirus.
Description
Technical Field
The invention provides a series of small molecule inhibitors designed based on the crystal structure of SARS coronavirus main protease, a preparation method and application thereof in preparing medicaments for treating or preventing various coronavirus infections.
Background
A novel coronavirus has been identified as a murder of Severe Acute Respiratory Syndrome (SARS) and is named SARS coronavirus (SARS-CoV). SARS Coronavirus belongs to the order of Nidovirales, the family of Coronaviridae, the genus of Coronaviridae, in the genus of genus species. It is a new variant of the coronavirus family, with a length of 29,736bp (Urbani Strain). SARS is a virulent infectious disease with a great threat to human beings, and no specific medicine or vaccine is available on the market at present.
According to the serological classification, the coronavirus family (coronavirus genus) includes four types (Group) in total. Type I includes porcine transmissible gastroenteritis virus (TGEV), human coronavirus (HCoV) strain 229E, Feline Infectious Peritonitis Virus (FIPV), and the like; type II includes bovine coronavirus (BCoV for short), Murine hepatitis virus (MHV for short), and the like; type III currently includes only three viruses, one of which is known as Avian Infectious Bronchitis Virus (AIBV); SARS-CoV is type IV. Various coronaviruses have great threat to the life health of people and livestock.
The genome of the SARS coronavirus encodes two large replicase polyproteins pp1a (486kDa) and pp1ab (790kDa), which are encoded by the genome comprising coronaviruses 2/3 to 3/4. These two proteins are hydrolyzed to produce many functional subunits of the viral replication complex. In this hydrolysis process, SARThe major proteolytic enzyme of S-coronavirus (main protease, abbreviated as M)pro33.8kDa, sometimes also referred to as 3C-Like Protein) plays a very critical role. The main protease, also a stretch of pp1a and pp1ab, is released by autocatalytic hydrolysis, which occurs at the Gln (Ser, Ala) site flanking the protease, by trans-splicing (bimolecular reaction). Under the action of the main protease, the replicase polyproteins pp1a and pp1ab are hydrolyzed into more than ten functional peptides, thereby further playing a role. If the hydrolysis of SARS coronavirus main protease can be inhibited, then the infection of SARS coronavirus to human body can be effectively resisted. Therefore, SARS coronavirus main protease is an ideal target for designing anti-SARS drugs.
Disclosure of Invention
It is an object of the present invention to provide a small molecule inhibitor that is effective in inhibiting the activity of the coronavirus main protease.
The invention also aims to provide a preparation method of the small molecule inhibitor.
It is a further object of the invention to provide the use of said small molecule inhibitor for the manufacture of a medicament for the treatment or prevention of a coronavirus infection.
Accordingly, the present invention provides a compound of the general formula (I):
wherein,
u is
OrWherein X is NH or CH2;R1Selected from the group consisting of: c3~C6Alkylcarbonyl, t-butoxycarbonyl, benzyloxycarbonyl, isoxazolylcarbonyl, furylcarbonyl, trifluoromethylcarbonyl,R2Selected from the group consisting of: c1~C4Alkyl, phenyl, benzyl, fluorobenzyl; r3Selected from the group consisting of: c1~C4Alkyl, phenyl, benzyl, p-methylphenyl, fluorobenzyl; r4Selected from the group consisting of: c1~C4Alkyl, phenyl, benzyl, p-methylphenyl, fluorobenzyl; r5Selected from the group consisting of: c1~C4Alkyl, phenyl, benzyl, p-methylphenyl, fluorobenzyl, fluorophenyl.
The invention also provides a preparation method of the compound of the general formula (I), which comprises the following steps:
the protecting group R of amino in the compound of formula (II)6Removing R therein6Selected from the group consisting of: t-butoxycarbonyl, trifluoroacetyl, benzyloxycarbonyl;
condensing the product of the last step with a compound of formula (III) in the presence of a condensing agent to obtain a compound of formula (I),
wherein R in the formula (II) and the formula (III)1、R2、R4And U is as defined for formula (I).
The invention also provides the use of a compound of formula (I) in the manufacture of a medicament for the treatment or prophylaxis of coronavirus infection.
Experiments prove that the compound can obviously inhibit the activity of main protease of coronavirus such as TGEV, HcoV, FIPV, AIBV, SARS-CoV and the like, and has good application prospect in the aspect of preparing medicaments for treating or preventing coronavirus infection.
Drawings
The drawings are not necessarily to scale, emphasis instead being placed upon better illustrating the present invention for the convenience of the reader. The invention may be better understood by considering the drawings in conjunction with the detailed description.
FIG. 1 shows small molecule N1 and SARS-CoV MproSurface map of monomer a binding;
FIG. 2 is a graph of electron density of N1 binding to the active pocket of monomer A;
FIG. 3 shows the small molecules N1, N2, N3 and N4 against SARS-CoV MproWherein SARS _3CL represents SARS coronavirus main protease, namely SARS-CoVMpro;
FIG. 4 shows the inhibitory activity curve of small molecule N1 against the transmissible gastroenteritis virus (TGEV) main protease (abbreviated as TGEV-3 CL in the figure);
FIG. 5 shows the inhibitory activity curves of small molecule N1 against human coronavirus 229E (HCoV) main protease (abbreviated as HCoV-3 CL in the figure);
FIG. 6 shows the inhibitory activity curve of small molecule N1 against Feline Infectious Peritonitis Virus (FIPV) major protease (abbreviated as FIPV _3CL in the figure);
FIG. 7 shows the inhibitory activity profile of small molecule N1 against Avian Infectious Bronchitis Virus (AIBV) main protease (abbreviated as AIBV _3CL in the figure).
Detailed Description
Definition of terms
For descriptive convenience, specific terminology is used herein and is explained below on a case-by-case basis.
"N1", "N2", "N3" and "N4" are particularly preferred small molecule inhibitors of the present invention having the structural formulas:
structural formula of N1
Structural formula of N2
Structural formula of N3
Structural formula of N4
As used herein, the terms "major proteolytic enzyme of SARS coronavirus", "SARS-CoV Mpro”、“SARS-CoV 3CLpro"," SARS coronavirus main protease "and the like, refer to the major proteolytic enzyme of SARS coronavirus.
“C3~C6Alkyl of (a) "means a straight or branched chain alkyl group having between 3 and 6 carbon atoms, including but not limited to: n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl and the like.
“C1~C4The "alkyl group" of (a) means a linear or branched alkyl group having between 1 and 4 carbon atoms, including: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, and the like.
"Fluorobenzyl" includes p-fluorobenzyl, m-fluorobenzyl, o-fluorobenzyl and the like.
"fluorophenyl" includes p-fluorophenyl, m-fluorophenyl, o-fluorophenyl, and the like.
In a partial structural formula, the following compounds,ipr represents isopropyl, Et represents ethyl, "Bn" represents benzyl, and "Boc" represents t-butyloxycarbonyl.
"TFA" represents trifluoroacetic acid, "THF" represents tetrahydrofuran, "DMF" represents N, N-dimethylformamide, "DMSO" represents dimethyl sulfoxide, "PhH" represents benzene, "iPr represents2NEt "stands for diisopropylethylamine," NEt3"represents triethylamine.
"DCC" represents dicyclohexylcarbodiimide (dicyclohexylcarbodiimide);
"DIEA" represents diisopropylethylamine (diisopropylethylamine);
"DMAP" represents 4-N, N-dimethylaminopyridine (4-N, N-dimethylammonylidine);
"EDCI" represents 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (1-ethyl-3- (3-dimethylamino propyl) carbodiimide hydrochloride);
"HATU" represents N- [ (dimethylamino) (3H-1, 2, 3-triazole (4, 5-b) pyridin-3-yloxy) methylene ] -N-methylbetaine hexafluorophosphate (N- [ (dimethyllamino) (3H-1, 2, 3-triazo (4, 5-b) pyridine-3-yloxy) methyl ] -N-methylethaneaminonium hexafluorophosphate;
"HBTU" represents O- (benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium hexafluorophosphate (O- (benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium hexafluorophosphate);
"HOBt" represents 1-hydroxybenzotriazole (1-hydroxybenzotriazole).
Structure of the Compound of the present invention
As mentioned above, the structure of the compound of the invention is shown in the general formula (I).
Certain preferred compounds of the invention correspond to the following formula,
further preferred compounds of the invention correspond to the following general formula,
further preferred compounds of the invention correspond to the following general formula,
some particularly preferred specific compounds of the invention are listed below,
process for the preparation of the compounds of the invention
The preparation method of the compound of formula (I) provided by the invention comprises the following steps:
the protecting group R of amino in the compound of formula (II)6Removing R therein6Selected from the group consisting of: t-butoxycarbonyl, trifluoroacetyl, benzyloxycarbonyl;
condensing the product of the last step with a compound of formula (III) in the presence of a condensing agent to obtain a compound of formula (I),
wherein R in the formula (II) and the formula (III)1、R2、R4And U is as defined for formula (I).
The synthesis of compounds of formula (II) can be referred to: qingping Tian, Naresh K.Nayyar, Srinivasan Babu, Lijian Chen, Junhua Tao, Steven Lee, Anthony Tibbetts, Terence Moran, Jason Liou, Ming Guo and Timothy P.Kennedy tetrahedron Lett.2001, 42, 6808-. The synthesis of compounds of formula (III) can be referred to: dawei Ma, Weiqing Xie, Bin Zou, Qiong Lei and Duanqing Pei Tetrahedron Lett.2004, 45, 8103-.
In some preferred embodiments of the present invention, the compound of formula (II) is reacted with an acid (e.g., a mixture of dichloromethane and trifluoroacetic acid at a volume ratio of 1-4: 1) in an organic solvent at room temperature for 1-4 hours; extracting the solvent, dissolving the compound of formula (III) in an aprotic solvent (e.g. CH)2Cl2,THF,CHCl3) Adding a condensing agent such as HATU, HBTU, EDCI, and adding an organic base such asiPr2NEt,NEt3And reacting at room temperature for 8-24 hours to obtain the compound shown in the formula (I).
R in the compounds of formula (I) may also be derivatized1Group exchange to the desired group R1', to give a series of derivatives of the compound of formula (I) wherein R1' is selected from the group consisting of: c3~C6Alkylcarbonyl, t-butoxycarbonyl, benzyloxycarbonyl, isoxazolylcarbonyl, furylcarbonyl, trifluoromethylcarbonyl,The derivatization comprises the following steps: r in the obtained compound of formula (I)1Removing; reacting the product of the previous step with a carboxylic acid R in the presence of a condensing agent1' -OH to give a derivatized compound of formula (I) wherein R1Radical is replaced by R1' group; wherein R is1' is selected from the group consisting of: c3~C6Alkylcarbonyl, t-butoxycarbonyl, benzyloxycarbonyl, isoxazolylcarbonyl, furylcarbonyl, trifluoromethylcarbonyl,
In some preferred embodiments of the present invention, the compound of formula (I) to be derivatized is reacted with an acid (e.g., a mixture of dichloromethane and trifluoroacetic acid at a volume ratio of 1-4: 1) in an organic solvent at room temperature for 1-4 hours; extracting the solvent with a carboxylic acid R1' -OH is dissolved in an aprotic solvent (e.g., CH)2Cl2,THF,CHCl3) Adding a condensing agent such as HATU, HBTU, EDCI, and adding an organic base such asiPr2NEt,NEt3And reacting at room temperature for 8-24 hours to obtain a series of derivatives of the compound shown in the formula (I).
Wherein the acid is preferably trifluoroacetic acid or hydrochloric acid; preferred organic solvents are selected from the group consisting of: CH (CH)2Cl2Tetrahydrofuran, tetrahydrofuran、CHCl3N, N-dimethylformamide, dioxane; preferred aprotic solvents are selected from the group consisting of: CH (CH)2Cl2Tetrahydrofuran, CHCl3N, N-dimethylformamide, dimethyl sulfoxide, benzene; preferred condensation reagents are selected from the group consisting of: HATU, HBTU, EDCI; preferred organic bases are selected from the group consisting of: diisopropylethylamine and triethylamine.
In order to explain the present invention in more detail, specific embodiments thereof will be given below with reference to the accompanying drawings. In describing these embodiments, well-known experimental methods, instruments, reagents, materials and the like have not been described in detail so as not to obscure the present invention.
Example 1
42mg ofDissolved in 2ml of CH2Cl20.5ml of TFA was added thereto, and the mixture was reacted at room temperature for 1 hour, and the solvent was then drained. The resulting de-Boc substrate was dissolved in 2ml of CH2Cl2In (1), 40mg ofThen 71 mul ofiPr2NEt, then 63mg of HATU. The reaction was carried out at room temperature for 12 hours, followed by 1M HCl and saturated NaHCO3Aqueous solution, saturated brine and Na2SO4And (5) drying. Filtering, evaporating solvent under reduced pressure, and performing flash column chromatography to obtain 52mg of productThe yield was 80%.
The spectrum data are as follows:
H NMR:δ(500MHz,CDCl3)0.76(d,3H,J=6.9Hz),0.98(d,3H,J=5.7Hz),1.30(t,3H,J=7.3Hz),1.42(s,9H),1.74-1.94(m,4H),2.10-2.40(m,2H),2.49-2.52(m,1H),2.67-2.71(m,1H),2.85-3.00(m,1H),3.05-3.17(m,1H),3.23-3.39(m,3H),4.18(q,2H,J=7.1Hz),4.41-4.51(m,1H),4.58-4.65(m,1H),5.03-5.10(m,1H),5.49(t,1H,J=14.7Hz),5.98(dd,1H,J=15.8Hz,4.4Hz),6.70-7.00(m,3H),7.12-7.16(m,3H);
ESI-MS:[M+H+]590.2,HRMS found m/z 612.3062,C31H44N3O7FNarequires 612.3065;
[α]D 24.0(c 0.69,CHCl3)。
example 2
Mixing 45mgDissolved in 2ml of CH2Cl20.5ml of TFA was added thereto, and the mixture was reacted at room temperature for 1 hour, and the solvent was then drained. The resulting de-Boc substrate was dissolved in 2ml of CH2Cl2In (1), 12mg ofThen 48. mu.l ofiPr2NEt, then 44mg of HATU. The reaction was carried out at room temperature for 12 hours, followed by 1M HCl and saturated NaHCO3Aqueous solution, saturated brine and Na2SO4And (5) drying. Filtering, evaporating the solvent under reduced pressure, and performing flash column chromatography to obtain 34mg of productThe yield was 73%.
The spectrum data are as follows:
H NMR:δ(300MHz,CDCl3)0.85(d,3H,J=6.6Hz),1.03(d,3H,J=6.9Hz),1.30(t,3H,J=7.2Hz),1.52-1.61(m,1H),1.71-1.90(m,2H),2.26-2.42(m,2H),2.48(s,3H),2.53-2.73(m,3H),2.84-2.98(m,2H),3.13-3.23(m,1H),3.30-3.42(m,2H),4.18(q,2H,J=7.2Hz),4.41-4.56(m,1H),4.66-4.76(m,1H),5.50(d,1H,J=15.6Hz),5.91(s,1H),6.39(s,1H),6.63(dd,1H,J=15.3Hz,4.8Hz),6.94-7.03(m,2H),7.11-7.35(m,4H);
ESI-MS:[M+H+]599.3,[M+Na+]621.3,HRMS found m/z 621.2717,C31H44N3O7FNa requires 621.2695;
[α]D 32.8(c 0.51,CHCl3)。
examples 1 and 2 can be briefly summarized by the following reaction schemes:
example 3
In describing this example, reference is made to the following reaction scheme (where the one or two digit arabic number below certain compounds is their numbering):
preparation of Compound 9
321mg (1mmol) of Compound 8 was dissolved in 2ml of dichloromethane, and 1ml of trifluoroacetic acid was added thereto, followed by stirring at room temperature for 1 hour. The solvent was evaporated under reduced pressure and pumped to dryness. The resulting product was dissolved in 4ml of dichloromethane and 260mg of the compound were added in order0.4ml ofDiisopropylethylamine, 162mg of HOBt, and finally 247mg of DCC were added, and the mixture was stirred at room temperature overnight. The reaction system was filtered and 10ml of dichloromethane was added, washed with 1M HCl, saturated aqueous sodium bicarbonate solution, and saturated aqueous sodium chloride solution in this order, dried over anhydrous sodium sulfate, filtered, the solvent was evaporated, and flash column chromatography was performed to obtain 387mg of compound 9 with a yield of 92%.
The spectrum data are as follows:
H NMR:δ(300MHz,CDCl3)0.90-0.95(m,12H),1.44(s,9H),1.45-1.68(m,3H),2.03-2.1 5(m,1H),3.88(dd,1H,J=6.6Hz,9.3Hz),4.64-4.72(m,1H),5.06(d,1H,J=9.9Hz),5.16(d,2H,J=3.3Hz),6.24(d,1H,J=10.5Hz),7.29-7.30(m,5H)。
preparation of Compound 10
331mg of Compound 9 was dissolved in 4ml of dichloromethane, and 1ml of trifluoroacetic acid was added thereto, followed by stirring at room temperature for 1 hour. The solvent was evaporated under reduced pressure and pumped to dryness. The resulting product was dissolved in 4ml of dichloromethane and 19mg of the compound were added in succession0.3ml of diisopropylethylamine, 128mg of HOBt, and finally 195mg of DCC were added and stirred at room temperature overnight. The reaction system was filtered, 10ml of dichloromethane was added, washed with 1M HCl, saturated aqueous sodium bicarbonate solution, and saturated aqueous sodium chloride solution in this order, dried over anhydrous sodium sulfate, filtered, the solvent was evaporated, and flash column chromatography was performed to obtain 357mg of compound 10 with a yield of 92%.
The spectrum data are as follows:
H NMR:δ(300MHz,CDCl3)0.89-0.94(m,12H),1.35(d,3H,J=7.2Hz),1.44(s,9H),1.56-1.69(m,3H),2.1 3-2.24(m,1H),4.09-4.19(m,1H),4.24(dd,1H,J=6.6Hz,8.7Hz),4.92-5.03(m,1H),4.95(d,2H,J=5.4Hz),6.35-6.44(m,1H),6.69-6.78(m,2H),7.31-7.42(m,5H)。
preparation of Compound 11
Dissolving 310mg of compound 10 in 5ml of methanol, adding 62mg of 20% palladium on carbon, hydrogenating at normal pressure for 3 hours, filtering off the palladium on carbon, evaporating the solvent, and carrying out flash column chromatography to obtain 258mg of compound 11 with a yield of 100%.
The spectrum data are as follows:
H NMR:δ(300MHz,CDCl3)0.9 1-0.96(m,12H),1.34(d,3H,J=6.9Hz),1.44(s,9H),1.60-1.75(m,3H),2.08-2.20(m,1H),4.1 8-4.28(m,1H),4.33(t,1H,J=9.0Hz),4.51-4.61(m,1H),5.27-5.35(m,1H),7.08-7.17(m,1H),7.27-7.33(m,1H)。
preparation of Compound N2
50mg ofDissolved in 2ml of CH2Cl20.5ml of TFA was added thereto, and the mixture was reacted at room temperature for 1 hour, and the solvent was then drained. The resulting de-Boc substrate was dissolved in 2ml of CH2Cl2To this, 62mg of Compound 11 was added, and 97. mu.l of the mixture was addediPr2NEt, then 75mg of HATU. The reaction was carried out at room temperature for 12 hours, followed by 1M HCl and saturated NaHCO3Aqueous solution, saturated brine and Na2SO4And (5) drying. Filtration, evaporation of the solvent under reduced pressure and flash column chromatography gave 76mg of product N2 in 81% yield.
The spectrum data are as follows:
ESI-MS:[M+H+]610.4。
preparation of Compound N1
41mg of Compound N2 were dissolved in 2ml of CH2Cl20.5ml of TFA was added thereto, and the mixture was reacted at room temperature for 1 hour, and the solvent was then drained. The resulting Boc-removed substrate was solubilizedTo 2ml of THF were added 10mg ofThen 42. mu.l ofiPr2NEt, then 33mg of HATU. The reaction was carried out at room temperature for 12 hours, followed by 1M HCl and saturated NaHCO3Washing the solution with saturated saline water, Na2SO4And (5) drying. Filtration, evaporation of the solvent under reduced pressure and flash column chromatography gave 34mg of product N1 in 82% yield.
The spectrum data are as follows:
H NMR:δ(300MHz,CDCl3)0.89(s,12H),1.25-1.29(m,3H),1.44(d,3H,J=7.4Hz),1.50-1.92(m,5H),2.04-2.17(m,2H),2.20-2.41(m,3H),2.47(s,3H),3.20-3.41(m,2H),4.18(q,2H,J=7.2Hz),4.32-4.43(m,1H),4.57-4.79(m,2H),4.85-4.97(m,1H),5.93-6.00(m,1H),6.46(s,1H),6.82-6.92(m,1H),7.47-7.93(m,3H);
ESI-MS:[M+H+]619.3。
EXAMPLE 4 SARS-CoV MproCrystal preparation, data collection and structure resolution of complexes with small molecule inhibitors
Materials and methods
1.SARS-CoV MproCrystal preparation of complexes with Small molecule Compound N1
SARS-CoV MproFurther isolation, purification and crystallization were carried out after expression in the E.coli strain BL21(DE3) (see: Yang H et al 2003.the Crystal Structures of SARSVirus Main Protease Mpro and Its Complex with an inhibitor. PNAS, 100 (23): 13190-. After N1 was dissolved in 10mM solution prepared from 7.5% PEG6000, 6% DMSO and 0.1M MES (pH6.0), the solution was added in an equal volume to the suspension of the grown crystals, and left to stand at 291K for 2 days.
2.SARS-CoV MproData collection and Structure of Complex with Small molecule Compound N1Parse
(1) Data collection and data processing
The data collection is completed by a Rigaku CuK alpha rotating target X-ray diffractometer, and the system parameters during data collection are voltage 40KV, current 20mA, wavelength 1.5418 Å and temperature 100K. The crystals were immersed in 30% PEG400, 0.1M MES (pH6.0) antifreeze solution and rapidly cooled to 100K with a stream of liquid nitrogen. The diffraction resolution of the crystal was 1.88 Å. All final diffraction data were treated with HKL2000 to 2.0 Å.
(2) Structural analysis
The structure of the complex is analyzed by using SARS-CoV MproThe parent structure (PDBcode: 1UJ1) of (2) was used as an initial search model, and the structure was analyzed by a molecular replacement method. A clear solution of symmetric disomy can be obtained by performing a rotation function and translation function search using CNS procedures. Then, a difference graph of 2Fo-Fc and Fo-Fc is used in the program O to build a small molecule model. After correction, Rwork20.0,Rfree23.8。
SARS-CoV MproThe methods for crystal preparation, data collection and structure analysis of the compound of N2, N3 and N4 are basically the same as those of N1, and are not described in detail herein.
Second, structural analysis
1、SARS-CoV MproStructural analysis of Complex with Small molecule Compound N1
SARS-CoV MproIs a homodimeric structure containing two monomers a and B in an asymmetric unit, since the binding of small molecule compound N1 to A, B two monomers is substantially similar, only the binding pattern of N1 and a is now analyzed.
In the 2Fo-Fc difference plot (counted at 1. sigma.) for monomer A, it was clearly observed that N1 was bound into the substrate binding pocket of the enzyme. Wherein C beta in N1 and S gamma of A145-Cys form a covalent bond with the bond length of 1.8 Å. The carbonyl oxygen in the N1 ester group and the amino group on the main carbon chain of A145-Cys form a hydrogen bond of 3.1 Å; the ethyl group in the ester forms a hydrophobic interaction with the side chains of A27-Leu, His-A41 and Thr-A25. At the P1 site, the oxygen in the five-membered ring of the lactam forms a 2.6 Å hydrogen bond with NE2 of His-163. One water molecule near the lactam ring can form hydrogen bonds with the carbonyl oxygen of N, His-172, NE2 and 140-Phe and Ser-B1 on the lactam ring of N1, respectively, to form 2.6 Å, 3.2 Å, 2.6 Å and 2.7 Å, so that the lactam ring is firmly bound in the S1 pocket. Since the P1 site of the main protease of the entire coronavirus family is well conserved and always appears as Q, occupying the corresponding substrate binding pocket S1 is a key to inhibiting protease activity. For the P2 site of N1, the Leu side chain of the small molecule was easily inserted into a hydrophobic pocket consisting of the His-41, Met-49 and Phe-181 side chains, and the alkyl portions of the Gln-189 and Asp-187 side chains. The carbonyl oxygen in His-164 forms a hydrogen bond of 2.9 Å with N in a peptide bond on the side close to the ester group in N1, the carbonyl oxygen of Val in N1 forms a hydrogen bond of 2.9 Å with N in Glu-A166, the N in a Val peptide bond forms a hydrogen bond of 3.0 Å with the Glu-A166 carbonyl oxygen, and the N in an Ala peptide bond in N1 forms a hydrogen bond of 3.2 Å with the carbonyl oxygen of Thr-190. The side chain of Ala is inserted into a hydrophobic pocket consisting of the side chains of Phe-185, Glu-192, Leu-167, Met-165. The terminal heterocycle in N1 has a hydrophobic interaction with the five-membered ring of Pro-168. All of these covalent, hydrogen bonding and hydrophobic interactions result in the inhibitor compound N1 being firmly bound within the active pocket of the enzyme substrate, thereby causing inactivation of the enzyme.
2. The combination of N2 and N3 is substantially similar to N1 and will not be described in detail.
Example 5 pairs of small molecule compounds N1, N2, N3 and N4 were derived from various sources
Inhibitory Activity of the Main protease of different coronaviruses
Materials and methods
(1) N1, N2, N3 and N4 for SARS-CoV MproMeasurement of inhibitory Activity of
To the buffer (20mM Tris-HCl pH7.0, 1mM DTT) was added 1. mu.M (final concentration) of SARS-CoV M pro100 μ M (final concentration)After compound N1, 298K was allowed to stand for 10 minutes, 5. mu.M of fluorescently labeled substrate (MCA-AVLQ ↓ SGFRL (DNP) L-NH2) was added rapidly. The excitation and emission wavelengths were 330nm and 395nm, respectively, and the temperature was maintained at 298K, and fluorescence readings were recorded every 2 seconds.
The inhibitory activities of N2, N3 and N4 were determined in substantially the same manner as N1.
Comparison: no inhibitor was added and the remaining conditions were the same. The results are shown in FIG. 3.
(2) Measurement of inhibitory Activity of N1 against Transmissible Gastroenteritis Virus (TGEV) Main protease.
mu.M of the protease, 100. mu.M of Compound N1, 298K, were added to a buffer (20mM Tris-HCl pH7.0, 1mM DTT) and after 10 minutes of standing, 5. mu.M of a fluorescently labeled substrate (MCA-AVLQSGFRL(DNP) L-NH) was rapidly added2). The excitation and emission wavelengths were 330nm and 395nm, respectively, and the temperature was maintained at 298K, and fluorescence readings were recorded every 2 seconds. The inhibitor concentration was then varied and the inhibitory activity was determined at 10nM, 1. mu.M, respectively. Comparison: no inhibitor was added and the remaining conditions were the same. The results are shown in FIG. 4.
(3) Determination of inhibitory Activity of N1 against Human Coronavir (HCoV) 229E Primary protease
mu.M of the protease, 100. mu.M of Compound N1, 298K, were added to a buffer (20mM Tris-HCl pH7.0, 1mM DTT) and after 10 minutes of standing, 5. mu.M of a fluorescently labeled substrate (MCA-AVLQSGFRL(DNP) L-NH2) was added rapidly. The excitation and emission wavelengths were 330nm and 395nm, respectively, and the temperature was maintained at 298K, and fluorescence readings were recorded every 2 seconds. Comparison: no inhibitor was added and the remaining conditions were the same. The results are shown in FIG. 5.
(4) Measurement of inhibitory Activity of N1 against major protease of Feline Infectious Peritonitis Virus (FIPV)
The procedure is as in (3), and the results are shown in FIG. 6.
(5) Determination of inhibitory Activity of N1 on Main protease of Avian Infectious Bronchitis Virus (AIBV)
mu.M of the protease, 100. mu.M of Compound N1, 298K, were added to a buffer (20mM Tris-HCl pH7.0, 1mM DTT) and after 10 minutes of standing, 5. mu.M of a fluorescently labeled substrate (MCA-AVLQSGFRL(DNP) L-NH2) was added rapidly. The excitation and emission wavelengths were 330nm and 395nm, respectively, and the temperature was maintained at 298K, and fluorescence readings were recorded every 2 seconds. Comparison: no inhibitor was added and the remaining conditions were the same. The results are shown in FIG. 7.
Expression purification references for FIPV, HCoV, TGEV: constellation of substripes specificities, ammonium coronavirus main proteins, J Gen Virol, 2002; 83(Pt 3): 595-9.
Second, results and analysis
1. N1, N2, N3 and N4 for SARS-CoV MproInhibiting activity of
As can be seen in FIG. 3, N1, N2, N3 and N4 are directed against SARS-CoV MproAll had inhibitory activity, and N1 had the strongest inhibitory activity.
2. Inhibitory Activity of N1 against other coronavirus Main proteases
From the measurement of the inhibitory activity of N1 on other coronavirus main proteases (i.e., FIG. 4-FIG. 7), it can be seen that N1 has inhibitory activity on TGEV, HcoV, FIPV and AIBV main proteases, wherein the inhibitory activity on TGEV main protease is strongest, and N1 still has inhibitory activity on TGEV main protease under the condition of 10 nM.
Since TGEV, HcoV and FIPV belong to the type I (serotype) of the family Coronaviridae, AIBV belongs to the type III and SARS-CoV belongs to the type IV, it is concluded that N1 has inhibitory activity against the major proteases of the entire family Coronaviridae.
References referred to herein, including patent documents, academic papers, publications, and the like, are hereby incorporated by reference in their entirety.
It should be noted that the various experimental procedures involved in the present invention are all conventional in the art, and if not specifically mentioned herein, those skilled in the art can refer to various common tool books, scientific literatures or related specifications, manuals, etc. before the filing date of the present invention.
Manufacturers, references, or detailed manufacturing methods for those particular or otherwise unavailable from the various experimental articles (including, but not limited to, chemical reagents, biological products, cells, organisms, instruments, etc.) referred to herein are noted; unless otherwise specified, are all conventional laboratory supplies and may be readily obtained by various means (e.g., purchased, self-prepared, etc.) prior to the filing date of the present application.
It will be understood by those skilled in the art that various changes and modifications in form and detail may be made therein without departing from the spirit and scope of the invention, and these are to be considered as falling within the scope of the invention.
Claims (12)
1. A compound of the following general formula (I):
wherein,
R1Selected from the group consisting of: c3~C6Alkylcarbonyl, t-butoxycarbonyl, benzyloxycarbonyl, isoxazolylcarbonyl, furylcarbonyl, trifluoromethylcarbonyl,
R2Selected from the group consisting of: c1~C4Alkyl, phenyl, benzyl, fluorobenzyl;
R3selected from the group consisting of: c1~C4Alkyl, phenyl, benzyl, p-methylphenyl, fluorobenzyl;
R4selected from the group consisting of: c1~C4Alkyl, phenyl, benzyl, p-methylphenyl, fluorobenzyl;
R5selected from the group consisting of: c1~C4Alkyl, phenyl, benzyl, p-methylphenyl, fluorobenzyl, fluorophenyl.
2. The compound of claim 1, wherein:
R1selected from the group consisting of: t-butyloxycarbonyl group, isoxazolylcarbonyl group, trifluoromethylcarbonyl group,
R2Is methyl or isopropyl;
R3is methyl or isopropyl;
R4is ethyl or benzyl;
R5selected from the group consisting of: isobutyl, p-fluorobenzyl, phenyl, fluorophenyl.
7. A process for the preparation of a compound according to claim 1, comprising the steps of:
the protecting group R of amino in the compound of formula (II)6Removing R therein6Selected from the group consisting of: t-butoxycarbonyl, trifluoroacetyl, benzyloxycarbonyl;
condensing the product of the last step with a compound of formula (III) in the presence of a condensing agent to obtain a compound of formula (I),
wherein R in the formula (II) and the formula (III)1、R2、R4And U is as defined for formula (I).
8. The method of claim 7, comprising the steps of:
in an organic solvent, reacting the compound of the formula (II) with an acid at room temperature for 1-4 hours, and removing the protecting group R of amino6Extracting the solvent, said R6Selected from the group consisting of: t-butoxycarbonyl, trifluoroacetyl, benzyloxycarbonyl;
and (3) dissolving the product and the compound of the formula (III) in an aprotic solvent, adding a condensation reagent and organic base, and reacting at room temperature for 8-24 hours to obtain the compound of the formula (I).
9. The process of claim 7, further comprising further derivatizing the resulting compound of formula (I) to convert R therein1Group exchange to the desired group R1', wherein R1' is selected from the group consisting of: c3~C6Alkylcarbonyl, t-butoxycarbonyl, benzyloxycarbonyl, isoxazolylcarbonyl, furylcarbonyl, trifluoromethylcarbonyl,
The derivatization comprises the following steps:
a process according to claim 7R in the compound of formula (I) obtained in (1)1Removing;
reacting the product of the previous step with a carboxylic acid R in the presence of a condensing agent1' -OH to give a derivatized compound of formula (I) wherein R1Radical is replaced by R1' group (a).
10. The method of any one of claims 7-9, wherein,
the acid is trifluoroacetic acid or hydrochloric acid;
the organic solvent is selected from the group consisting of: CH (CH)2Cl2Tetrahydrofuran, CHCl3N, N-dimethylformamide, dioxane;
the aprotic solvent is selected from the group consisting of: CH (CH)2Cl2Tetrahydrofuran, CHCl3N, N-dimethylformamide, dimethyl sulfoxide, benzene;
the condensing agent is selected from the group consisting of: HATU, HBTU, EDCI;
the organic base is selected from the group consisting of: diisopropylethylamine and triethylamine.
11. Use of a compound according to claims 1-6 in the manufacture of a medicament for the treatment or prevention of a coronavirus infection.
12. Use according to claim 11, wherein the coronavirus is a SARS coronavirus, a transmissible gastroenteritis virus of swine, a human coronavirus, a feline infectious peritonitis virus or an avian infectious bronchitis virus.
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CNB2004100877217A CN100363346C (en) | 2004-10-22 | 2004-10-22 | Small molecular inhibiting agent for coronavirus main proteinase, its preparation method and uses |
PCT/CN2005/001748 WO2006042478A1 (en) | 2004-10-22 | 2005-10-24 | Small-molecule inhibitors of coronaviral main protease, their preparation and use |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CNB2004100877217A CN100363346C (en) | 2004-10-22 | 2004-10-22 | Small molecular inhibiting agent for coronavirus main proteinase, its preparation method and uses |
Publications (2)
Publication Number | Publication Date |
---|---|
CN1763002A CN1763002A (en) | 2006-04-26 |
CN100363346C true CN100363346C (en) | 2008-01-23 |
Family
ID=36202685
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CNB2004100877217A Expired - Fee Related CN100363346C (en) | 2004-10-22 | 2004-10-22 | Small molecular inhibiting agent for coronavirus main proteinase, its preparation method and uses |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN100363346C (en) |
WO (1) | WO2006042478A1 (en) |
Families Citing this family (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103044338B (en) * | 2012-12-12 | 2016-08-03 | 天津医科大学总医院 | MiR-21 micromolecular inhibitor and application |
CN104592349A (en) * | 2015-02-15 | 2015-05-06 | 天津国际生物医药联合研究院 | Small molecule inhibitor of coronavirus main proteases as well as preparation method and application of small molecule inhibitor |
CN105412151A (en) * | 2015-12-11 | 2016-03-23 | 天津国际生物医药联合研究院 | Inhibitor aiming at coronavirus main protease, and application of Zn<2+> |
CN105837487A (en) * | 2016-03-17 | 2016-08-10 | 天津国际生物医药联合研究院 | Small-molecule inhibitor against MERS-CoV main protease, and preparation method and application thereof |
CA3024120A1 (en) * | 2016-05-13 | 2017-11-16 | Emory University | Peptidomimetics for the treatment of norovirus infection |
CN115645396A (en) * | 2020-02-26 | 2023-01-31 | 上海科技大学 | Application of propyl-2-imidazole disulfide in resisting coronavirus |
EP3912628A1 (en) * | 2020-05-20 | 2021-11-24 | Institut de Recherche en Semiochimie et Ethologie Appliquée | Nucleoside analogues to inhibit the main protease of a coronavirus |
US11351149B2 (en) * | 2020-09-03 | 2022-06-07 | Pfizer Inc. | Nitrile-containing antiviral compounds |
CN114149415A (en) * | 2021-07-26 | 2022-03-08 | 中国药科大学 | Peptide-like compound and derivative, preparation method, pharmaceutical composition and application thereof |
CN117209558B (en) * | 2023-08-14 | 2024-09-17 | 北京大学深圳研究生院 | Use of Mpro protein inhibitors in medicaments for the treatment of novel coronavirus infections |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998043950A1 (en) * | 1997-03-28 | 1998-10-08 | Agouron Pharmaceuticals, Inc. | Antipicornaviral compouds, compositions containing them, and methods for their use |
EE200000621A (en) * | 1998-04-30 | 2002-06-17 | Agouron Pharmaceuticals, Inc. | Anti-picornaviral compounds, their preparation and use |
US6355807B1 (en) * | 1999-08-24 | 2002-03-12 | Agouron Pharmaceuticals, Inc. | Efficient synthetic routes for the preparation of rhinovirus protease inhibitors and key intermediates |
CA2550515A1 (en) * | 2003-12-31 | 2005-07-21 | Taigen Biotechnology | Protease inhibitors |
-
2004
- 2004-10-22 CN CNB2004100877217A patent/CN100363346C/en not_active Expired - Fee Related
-
2005
- 2005-10-24 WO PCT/CN2005/001748 patent/WO2006042478A1/en active Application Filing
Non-Patent Citations (1)
Title |
---|
an efficinet and runabler oute to ag 70888 a rhinovirusproteaseinhi bitor. dawet ma et al.Tetrahedron letters,Vol.45 . 2004 * |
Also Published As
Publication number | Publication date |
---|---|
CN1763002A (en) | 2006-04-26 |
WO2006042478A1 (en) | 2006-04-27 |
WO2006042478A8 (en) | 2006-07-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP1012180B1 (en) | Hepatitis c inhibitor peptide analogues | |
US4644055A (en) | Method for preparing specific inhibitors of virus-specified proteases | |
Ahn et al. | Peptidomimetics and peptide backbone modifications | |
WO2006042478A1 (en) | Small-molecule inhibitors of coronaviral main protease, their preparation and use | |
CN1312817A (en) | Dolastatin 15 derivatives | |
WO2005118620A2 (en) | Methods for preparing internally constraied peptides and peptidomimetics | |
CA2418204A1 (en) | Novel peptides as ns3-serine protease inhibitors of hepatitis c virus | |
EP1003775A2 (en) | Hepatitis c inhibitor peptides | |
MacDonald et al. | Approaches to cyclic peptide beeta turn mimics | |
CN104703997A (en) | Solution phase processes for the manufacture of macrocyclic depsipeptides and new intermediates | |
US12084476B2 (en) | Tailored cyclodepsipeptides as potent non-covalent serine protease inhibitors | |
Majer et al. | Structure‐based subsite specificity mapping of human cathepsin D using statine‐based inhibitors | |
JP3325017B2 (en) | Novel metalloprotease inhibitors, their therapeutic use and methods for preparing starting compounds in their synthesis | |
Vaz et al. | Stable helical peptoids via covalent side chain to side chain cyclization | |
WO2022145444A1 (en) | Method for supporting amino acid on resin for solid-phase synthesis | |
WO2001096540A2 (en) | Hepatitis c protease exosite for inhibitor design | |
JPH03215497A (en) | Anti-herpes pentapeptide derivative having side chain of substituted aspartic acid | |
Naka et al. | Evaluations of substrate specificity and inhibition at PR/p3 cleavage site of HTLV-1 protease | |
Ekkati et al. | Preparation of N-acetyl, tert-butyl amide derivatives of the 20 natural amino acids | |
Rajesh et al. | α-Dehydro β-amino acid derivatives as turn inducer: Synthesis of potential HIV protease inhibitors based on structural mimicry | |
JP4485048B2 (en) | Prolyl endopeptidase inhibitory peptide | |
JPH0794418B2 (en) | New manufacturing method | |
KR20220102699A (en) | Peptide comprising stabilized polyproline Ⅱ helix | |
CN118063550A (en) | Novel brucella-resistant polypeptide and preparation method and application thereof | |
JPS63145299A (en) | Tetrapeptide methyl ketone inhibitor of virus protease |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
C17 | Cessation of patent right | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20080123 Termination date: 20101022 |