CN103183631B - Isatin-5-sulfonic acid amide derivatives and the application in the medicine of preparation treatment severe acute respiratory syndrome thereof - Google Patents
Isatin-5-sulfonic acid amide derivatives and the application in the medicine of preparation treatment severe acute respiratory syndrome thereof Download PDFInfo
- Publication number
- CN103183631B CN103183631B CN201110446695.2A CN201110446695A CN103183631B CN 103183631 B CN103183631 B CN 103183631B CN 201110446695 A CN201110446695 A CN 201110446695A CN 103183631 B CN103183631 B CN 103183631B
- Authority
- CN
- China
- Prior art keywords
- isatin
- compound
- sars
- medicine
- sulfonic acid
- 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
- 239000003814 drug Substances 0.000 title claims abstract description 13
- 238000011282 treatment Methods 0.000 title claims abstract description 5
- 201000003176 Severe Acute Respiratory Syndrome Diseases 0.000 title claims description 21
- KHHJNWWJDYSSRK-UHFFFAOYSA-N 2,3-dioxo-1h-indole-5-sulfonamide Chemical class NS(=O)(=O)C1=CC=C2NC(=O)C(=O)C2=C1 KHHJNWWJDYSSRK-UHFFFAOYSA-N 0.000 title abstract description 4
- 238000002360 preparation method Methods 0.000 title abstract description 3
- 150000001875 compounds Chemical class 0.000 claims abstract description 31
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims abstract description 3
- 241000315672 SARS coronavirus Species 0.000 claims description 8
- 238000004519 manufacturing process Methods 0.000 claims 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims 1
- 229910052739 hydrogen Inorganic materials 0.000 claims 1
- 239000001257 hydrogen Substances 0.000 claims 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Substances N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 5
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 4
- 206010003757 Atypical pneumonia Diseases 0.000 abstract description 3
- 208000015181 infectious disease Diseases 0.000 abstract description 2
- 230000002458 infectious effect Effects 0.000 abstract description 2
- 238000010189 synthetic method Methods 0.000 abstract description 2
- 239000003205 fragrance Substances 0.000 abstract 1
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 18
- JXDYKVIHCLTXOP-UHFFFAOYSA-N isatin Chemical compound C1=CC=C2C(=O)C(=O)NC2=C1 JXDYKVIHCLTXOP-UHFFFAOYSA-N 0.000 description 13
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- 108010055591 SARS coronavirus 3C-like protease Proteins 0.000 description 11
- 238000006243 chemical reaction Methods 0.000 description 9
- 239000012634 fragment Substances 0.000 description 9
- 229940079593 drug Drugs 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- 238000000034 method Methods 0.000 description 8
- 239000007787 solid Substances 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 7
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 6
- 229960000723 ampicillin Drugs 0.000 description 6
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 6
- 238000005160 1H NMR spectroscopy Methods 0.000 description 5
- -1 isatin compound Chemical class 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 239000013598 vector Substances 0.000 description 5
- 230000002401 inhibitory effect Effects 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 101800000535 3C-like proteinase Proteins 0.000 description 3
- 101800002396 3C-like proteinase nsp5 Proteins 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 101800000508 Non-structural protein 5 Proteins 0.000 description 3
- 108091005804 Peptidases Proteins 0.000 description 3
- 239000004365 Protease Substances 0.000 description 3
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- 238000001976 enzyme digestion Methods 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 235000019419 proteases Nutrition 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 230000001131 transforming effect Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 229940125673 3C-like protease inhibitor Drugs 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 241000620209 Escherichia coli DH5[alpha] Species 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 2
- 239000011543 agarose gel Substances 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 230000005496 eutectics Effects 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000012046 mixed solvent Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 2
- 239000011541 reaction mixture Substances 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- GHYOCDFICYLMRF-UTIIJYGPSA-N (2S,3R)-N-[(2S)-3-(cyclopenten-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-3-hydroxy-3-(4-methoxyphenyl)-2-[[(2S)-2-[(2-morpholin-4-ylacetyl)amino]propanoyl]amino]propanamide Chemical compound C1(=CCCC1)C[C@@H](C(=O)[C@@]1(OC1)C)NC([C@H]([C@@H](C1=CC=C(C=C1)OC)O)NC([C@H](C)NC(CN1CCOCC1)=O)=O)=O GHYOCDFICYLMRF-UTIIJYGPSA-N 0.000 description 1
- KEQGZUUPPQEDPF-UHFFFAOYSA-N 1,3-dichloro-5,5-dimethylimidazolidine-2,4-dione Chemical compound CC1(C)N(Cl)C(=O)N(Cl)C1=O KEQGZUUPPQEDPF-UHFFFAOYSA-N 0.000 description 1
- NQAVEPSYAPOTOQ-UHFFFAOYSA-N 1-methyl-5-(4-methylpiperazin-1-yl)sulfonylindole-2,3-dione Chemical compound C=1C=C2N(C)C(=O)C(=O)C2=CC=1S(=O)(=O)N1CCN(C)CC1 NQAVEPSYAPOTOQ-UHFFFAOYSA-N 0.000 description 1
- KIHNTPDWZLWMNO-UHFFFAOYSA-N 2,3-dioxoindole-1-sulfonamide Chemical class C1=CC=C2N(S(=O)(=O)N)C(=O)C(=O)C2=C1 KIHNTPDWZLWMNO-UHFFFAOYSA-N 0.000 description 1
- RUHJZSZTSCSTCC-UHFFFAOYSA-N 2-(bromomethyl)naphthalene Chemical compound C1=CC=CC2=CC(CBr)=CC=C21 RUHJZSZTSCSTCC-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
- 108010091324 3C proteases Proteins 0.000 description 1
- ZVAIPJCURHADPA-UHFFFAOYSA-N 5-(4-Methylpiperazin-1-yl)sulfonyl-1H-indole-2,3-dione Chemical compound C1CN(C)CCN1S(=O)(=O)C1=CC=C(NC(=O)C2=O)C2=C1 ZVAIPJCURHADPA-UHFFFAOYSA-N 0.000 description 1
- RQCUTXHKWJPTCR-UHFFFAOYSA-N 5-(4-pyridin-2-ylpiperazin-1-yl)sulfonyl-1h-indole-2,3-dione Chemical compound C1=C2C(=O)C(=O)NC2=CC=C1S(=O)(=O)N(CC1)CCN1C1=CC=CC=N1 RQCUTXHKWJPTCR-UHFFFAOYSA-N 0.000 description 1
- MBVCESWADCIXJN-UHFFFAOYSA-N 5-Bromoisatin Chemical compound BrC1=CC=C2NC(=O)C(=O)C2=C1 MBVCESWADCIXJN-UHFFFAOYSA-N 0.000 description 1
- LVNMAAGSAUGNIC-BQBZGAKWSA-N Cys-His Chemical compound SC[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CNC=N1 LVNMAAGSAUGNIC-BQBZGAKWSA-N 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
- 241001198387 Escherichia coli BL21(DE3) Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 241000430519 Human rhinovirus sp. Species 0.000 description 1
- 241001454768 Mentzelia nuda Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-diisopropylethylamine Substances CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 1
- RFWOJOUGYLVELN-UHFFFAOYSA-N O=C(C(N(CC1)CCN1c1cc(C(F)(F)F)ccc1)=O)c(cc1C2=O)ccc1NC2=O Chemical compound O=C(C(N(CC1)CCN1c1cc(C(F)(F)F)ccc1)=O)c(cc1C2=O)ccc1NC2=O RFWOJOUGYLVELN-UHFFFAOYSA-N 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- 101710200092 Replicase polyprotein Proteins 0.000 description 1
- 108030001409 SARS coronavirus main proteinases Proteins 0.000 description 1
- 102000012479 Serine Proteases Human genes 0.000 description 1
- 108010022999 Serine Proteases Proteins 0.000 description 1
- KEAYESYHFKHZAL-UHFFFAOYSA-N Sodium Chemical compound [Na] KEAYESYHFKHZAL-UHFFFAOYSA-N 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 150000001413 amino acids Chemical group 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000005571 anion exchange chromatography Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 125000004196 benzothienyl group Chemical group S1C(=CC2=C1C=CC=C2)* 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 125000005392 carboxamide group Chemical group NC(=O)* 0.000 description 1
- NWSBNVVOFKKFNV-UHFFFAOYSA-N chloroform;oxolane Chemical compound ClC(Cl)Cl.C1CCOC1 NWSBNVVOFKKFNV-UHFFFAOYSA-N 0.000 description 1
- XTHPWXDJESJLNJ-UHFFFAOYSA-N chlorosulfonic acid Substances OS(Cl)(=O)=O XTHPWXDJESJLNJ-UHFFFAOYSA-N 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 229940125904 compound 1 Drugs 0.000 description 1
- 229940125782 compound 2 Drugs 0.000 description 1
- 229940126214 compound 3 Drugs 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000009510 drug design Methods 0.000 description 1
- 238000009112 empiric therapy Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 239000003862 glucocorticoid Substances 0.000 description 1
- 125000005843 halogen group Chemical group 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 125000000623 heterocyclic group Chemical group 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 125000001165 hydrophobic group Chemical group 0.000 description 1
- 239000005457 ice water Substances 0.000 description 1
- 239000002955 immunomodulating agent Substances 0.000 description 1
- 229940121354 immunomodulator Drugs 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 108010026228 mRNA guanylyltransferase Proteins 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 125000002757 morpholinyl group Chemical group 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
- 125000001624 naphthyl group Chemical group 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
- 239000012074 organic phase Substances 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 125000004193 piperazinyl group Chemical group 0.000 description 1
- 125000003386 piperidinyl group Chemical group 0.000 description 1
- GZRKXKUVVPSREJ-UHFFFAOYSA-N pyridinylpiperazine Chemical compound C1CNCCN1C1=CC=CC=N1 GZRKXKUVVPSREJ-UHFFFAOYSA-N 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 208000023504 respiratory system disease Diseases 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 239000012312 sodium hydride Substances 0.000 description 1
- 229910000104 sodium hydride Inorganic materials 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 229940124530 sulfonamide Drugs 0.000 description 1
- 238000009120 supportive therapy Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 229940037128 systemic glucocorticoids Drugs 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000003041 virtual screening Methods 0.000 description 1
- 239000009828 yingliu Substances 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to isatin 5 sulfonic acid amide derivatives shown in Formulas I:Wherein, R1Selected from C1‑C6Alkyl, fragrance methyl, heterocycle arylmethyl；R2For nitrogen heterocycle.The invention still further relates to synthetic method and its application in the medicine of preparation treatment infectious atypical pneumonia of compound of formula I.
Description
Technical Field
The invention relates to novel isatin-5-sulfonamide derivatives, a synthetic method of the compounds and application of the compounds in preparing a medicament for treating infectious atypical pneumonia.
Background
SARS is a highly contagious Respiratory disease called Severe Acute Respiratory Syndrome (SARS) by the world health organization. SARS is mainly spread by close-range air entrainment and close contact, and clinically, it is mainly manifested as pneumonia, and there is a significant accumulation phenomenon in homes and hospitals. In 2003, SARS matting rolled up 32 countries worldwide, causing a tremendous panic, and the warriors of this storm started to be SARS coronavirus.
For a long time, there have been no clinically proven therapeutically specific drugs available to both humans and animals. During the SARS outbreak in 2003, only empirical therapy, supportive therapy and treatment methods to control complications have been available clinically. Among them, antiviral drugs, glucocorticoids, antibiotics and immunomodulators are used in large quantities. Although playing an important role in some stages, the medicines have certain defects, and bring certain interference to the research of various physical signs of SARS patients and the observation of the normal pathogenesis of the SARS patients. In 2003, the roe and academy laboratories successfully analyzed the eutectic structure of SARS major protease at different pH values and a single inhibitor (hexapeptidyl CMK), the enzyme is generalized serine protease with Cys-His as the active site, and the replication and transcription of the virus are regulated by hydrolyzing replicase polyprotein to release replicase PP1a and PP1b, which provides a greater possibility for developing specific and effective anti-SARS virus drugs.
In general, no specific drug for SARS has been tested clinically. Although SARS is effectively controlled worldwide, the human cannot ensure that SARS storm does not cause heavy soil scaring, and the development of high-efficiency anti-SARS drugs aiming at the existing research results is of great significance in order to reduce the loss of life and property.
After an outbreak of SARS, the development of therapeutic drugs for this has been widely spread worldwide. According to the research of virus protease inhibitor with similar structure with SARS protein main protease and computer aided virtual screening, various chemical structures have been found to produce SARS inhibiting effect. Of these, isatin is the most attractive one. In 2005, Chen topic group [ Chen, l. -r.; wang, Y. -C., etc.; the study of bioorg.med.chem.lett.2005,15,3058.] gave that the most active isatin compound was 1- (2-benzothiophenemethylene) -5-iodo-2, 3-dioxoindoline (compound 1), with an IC50 of 0.95 μmol L-1; in 2007, a preliminary study of the Liu subject group [ Lu Zhou, YingLiu, etc.; j.med.chem.2006,49,3440-3443 ] is 1- (6-naphthylmethylene) -2, 3-dioxoindoline-5-carboxamide (compound 2), IC50 is 0.37 μmol L-1; after further study, the subject group obtained a novel SARS coronavirus 3CL protease inhibitor 1- (2-benzothiophenemethylene) -2, 3-dioxoindoline-5-carboxamide (Compound 3) whose IC50 was (0.76. + -. 0.02). mu. mol L-1. These results indicate that the 5-position of isatin is a carboxamide group superior to a halogen atom, and that the activity is increased by a large hydrophobic group such as a naphthyl group, benzothiophenyl group, etc. at the 1-position. Meanwhile, the isatin structure is also proved to be a good mother nucleus structure for researching SARS virus resisting medicines.
However, the above compounds still have various problems, and neither of the above documents describes the compounds of the present invention, and there is no suggestion as to whether or not the 5-position sulfonamide compounds have SARS 3CL inhibitory activity. Therefore, there is a need for further research on derivatives of isatin, and the structure of isatin is improved, so as to obtain more effective drugs.
Disclosure of Invention
The invention takes the eutectic structure of SARS coronavirus main protein Nsp5 and 5-bromoisatin as the initial basis of research, synthesizes a series of isatin sulfonamide compounds which are not reported in literature through computer-aided drug design, and screens the compounds for inhibiting SARS protein Nsp5, and data shows that the compounds have certain inhibition effect and can be used as potential SARS coronavirus main protein Nsp5 inhibitors.
The compounds of the present invention have the structure shown in formula I below:
wherein,
R1optional C1-C6Aromatic methyl, heterocyclic aromatic methyl, for example:
n is an integer of 0 to 6.
R2Are nitrogen-containing heterocyclic groups including piperazine groups, morpholinyl, piperidinyl, and the like, for example:
the compound of the formula I can be synthesized by the method shown in the following reaction formula:
the compounds of the present invention will be explained and verified in detail by specific examples below.
Detailed Description
The following examples are presented to enable those skilled in the art to more fully understand the present invention and are not intended to limit the invention in any way.
The first embodiment is as follows: 1- (2-naphthylmethyl) -5- [4- (2-pyridyl) piperazine-1-sulfonyl]Isatin (d)1)
The method comprises the following specific steps:
13.0ml of chlorosulfonic acid was added dropwise to the isatin in an ice bath under a nitrogen blanket in a 2.94g ice bath. The above reactants were heated to 70 ℃ and reacted for 3 hours. The reaction solution was cooled to room temperature, slowly poured into 100g of ice, and the resulting yellow solid was filtered and washed with ice water, and then the resulting solid was dissolved with ethyl acetate, dried over anhydrous sodium sulfate, and the solvent was removed to give a yellow oily liquid a. The next reaction was carried out without purification.
Dissolving the product a 601mg obtained in the first step in 6ml of chloroform-tetrahydrofuran mixed solvent with the volume ratio of 1:1, cooling to 0 ℃ under the protection of nitrogen, dripping 1ml of chloroform dissolved with 1.117g of 1- (2-pyridyl) piperazine, 532 mu l N and N-diisopropylethylamine into the mixture, reacting and stirring overnight at room temperature, removing the solvent to obtain brown oily liquid b, and carrying out the next reaction without purification.
Dissolving the product b in the previous step in 15ml of mixed solvent of acetic acid and water mixture with the volume ratio of 1:1, and heating to 96 ℃ for reaction for 20 hours. After the reaction was cooled, 11g of sodium bicarbonate was slowly added, and the product c was extracted three times with ethyl acetate, washed with water, dried over magnesium sulfate, and purified by column chromatography (eluent: dichloromethane: methanol: 100: 1).
Dissolving 1mmol of the product c in anhydrous 5ml of anhydrous dimethylformamide, adding 1.5mmol of sodium hydride, reacting at 0 ℃ for 15min, dissolving 1.5mmol of 2-bromomethylnaphthalene in 1ml of dimethylformamide, dripping into the reaction mixture, reacting at room temperature for 3 hours, extracting with ethyl acetate and water, collecting an organic phase, removing the solvent, and purifying by a column (eluent: ethyl acetate: petroleum ether ═ 2:1) to obtain a yellow solid d, namely the compound of the embodiment.
Yellow solid, yield 75%, 1H NMR (400MHz, DMSO, in ppm) 3.00(t, J ═ 8.2Hz,4H),3.62(t, J ═ 8.8Hz,4H),5.12(s,2H),6.70(t, J ═ 8Hz,1H),6.91(d, J ═ 8Hz,1H),
7.207(d,J＝8.4Hz,1H)，7.50(m,2H),7.62(m,2H)，7.82(m,2H),7.91(m,3H),8.01(s,1H),8.04(d,J＝6.4Hz,1H)；ESI-MS m/z513.20([M+H+])
example two: 5- [4- (2-pyridinyl) piperazine-1-sulfonyl ] isatin
The following compound (c) was synthesized by selecting the corresponding starting materials in the same manner as in the synthesis of Compound 12)：
Yellow solid, yield 80%, 1H NMR (400MHz, CDCl)3,in ppm):2.98(t,J＝9.6Hz,4H),3.60(t,J＝8.8Hz,4H),6.63(m,1H),6.80(d,J＝8.8Hz,1H),7.13(dJ＝8Hz,1H),
7.5(m,1H),7.70(d,J＝2Hz,1H),7.92(d,J＝10Hz,1H),8.06(d,J＝6.4Hz,1H),
11.56(s,1H)；ESI-MS m/z 373.13([M+H+]),371.19([M-H+])
Example three: 5- (4-methylpiperazine-1-sulfonyl) isatin (c)3)
Yellow solid, yield 83%, 1H NMR (400MHz, CDCl3, in ppm): 2.15(s,3H),2.37(t, J ═ 8.8Hz,4H),2.90(s,4H),7.12(d, J ═ 8.4Hz,1H),7.68(d, J ═ 1.6Hz,1H),7.91(b, J ═ 10Hz,1H),11.47(s, 1H); ESI-MSm/z310.15([ M + H ]+]),308.20([M-H+])
Example four: 1-methyl-5- (4-methylpiperazine-1-sulfonyl) isatin (d)4)
Yellow solid, yield 73%, 1H NMR (400MHz, DMSO, in ppm): 2.13(s,3H),2.74(t, J ═ 8.8Hz,4H),2.93(s,4H),3.21(s,3H),7.38(d.J ═ 8.4Hz,1H),7.74(s,1H),8.02(d, J ═ 10Hz, 1H); ESI-MSm/z324.20([ M + H ]+])
Example five: 5- [4- (3-trifluoromethyl) phenylpiperazine-1-sulfonyl]Isatin (c)5)
Yellow solid, yield 80%, 1H NMR (400MHz, DMSO, in ppm): 3.05(t, J ═ 9.2Hz,4H),3.33(t, J ═ 12.8Hz,4H),7.15(m,4H),7.41(t, J ═ 1.6Hz,1H),7.73(d, J ═ 2Hz,1H),7.963(t, J ═ 8.4Hz,1H),11.48(s, 1H); ESI-MSm/z440.19([ M + H ]+]),438.22([M-H+])
The pharmacological activity test method is as follows:
expression and purification of SARS coronavirus main protease
Expression and purification of the SARS coronavirus main protease were performed according to the literature (Yang H et al Proc Natl Acad Sci.2003 Nov; 100(23): 13190-5). The specific method comprises the following steps:
1.1 the construction of the expression vector of SARS coronavirus main protease, which comprises the following steps:
a. the cDNA library of SARS virus strain numbered BJ01 provided by Beijing Hua large gene center is used for in vitro amplification by PCR technology;
a forward primer: 5'-CGGGATCCAGTGGTTTTAGGAAAATG-3'
Reverse primer: 5'-CCGCTCGAGTCATTGGAAGGTAACACCAGA-3'
b. After the gene fragment amplified by PCR is cut by BamHI and XhoI double enzymes, the fragment with the size of about 1kb is recovered by agarose gel electrophoresis;
c. connecting the recovered fragment with a T vector, then transforming Escherichia coli DH5 alpha competent cells with the connection product, coating the competent cells on an LB plate (containing 100mg/L ampicillin), and culturing overnight;
d. a plurality of single colonies were picked up from the plate, inoculated into tubes containing about 5mL of LB (ampicillin was added to the LB solution so that the final concentration was 100mg/L), and cultured overnight. Then extracting plasmids by using a plasmid extraction kit (B type plasmid small-quantity rapid extraction kit of Boda Taike company), carrying out enzyme digestion by using BamHI and XhoI, and then recovering a target gene fragment with the size of about 1kb by using agarose gel;
e. the target vector pGEX-4T-1 (purchased from Pharmacia) was digested with BamHI and XhoI, and then the digested fragment was recovered by agarose gel;
f. and (3) connecting the fragments obtained in the step (d) and the fragment obtained in the step (e) (mixing the target gene fragment and the target vector fragment which are recovered by enzyme digestion according to the molar ratio of 3: 1-6: 1, reacting for 30 minutes-18 hours at 16 ℃ according to the requirement of Takara DNA Ligation), transforming the competent cells of the Escherichia coli DH5 alpha, and coating the competent cells on an LB plate (containing 100mg/L ampicillin) for overnight culture. The positive clones screened were used for identification and sequencing. The sequencing result shows that the coding gene of the main protease of SARS coronavirus has been correctly cloned into pGEX-4T-1 vector.
1.2 expression and purification of SARS coronavirus main protease, which comprises the following steps:
a. transforming Escherichia coli BL21(DE3) strain with pGEX-4T-1 vector containing SARS coronavirus main protease gene obtained in the step 1.1, and screening positive clone with LB plate (containing 100mg/L ampicillin);
b. positive clones (single colonies grown on LB plates containing ampicillin) were picked up on LB plates as described in a, cultured overnight, and then transferred to 1L of LB medium (containing 100mg/L ampicillin) when OD was reached600Adding about 1mM IPTG when the concentration reaches 0.6-0.8, and culturing at 16 deg.C for about 12 hr;
centrifuging at 5000-8000 rpm for 10-15 min to collect cells, and then ultrasonically breaking the bacteria in ice bath for 20-30 min; centrifuging the bacterium breaking solution at 13000-15000 rpm for 20-40 min, and collecting the supernatant;
d. the supernatant was added to a GST affinity column (GE) pre-equilibrated with PBS, and 20-30 bed volumes were eluted with PBS to remove contaminating proteins. Finally, adding about 2mL of human rhinovirus 3C protease with the concentration of about 0.1mg/mL, carrying out enzyme digestion at 4 ℃ for 12-20 hours, and then collecting protein;
e. and (3) purifying the protein obtained in the last step d by using Mono Q (GE company) anion exchange chromatography to obtain the SARS coronavirus main protease with higher purity.
Screening method of SARS coronavirus main protease inhibitor
The method for screening SARS coronavirus main protease inhibitor adopted by the invention is a screening method disclosed in CN101418334A, and the specific method is as follows:
the activity of SARS coronavirus main protease was measured by using the fluorogenic substrate MCA-AVLQSGFR-Lys (Dnp) -Lys-NH2 (purity: 95% or more, Gill Biochemical Co., Ltd., Shanghai). The amino acid sequence of the fluorescent substrate is derived from the N-terminal self-cleavage sequence of SARS coronavirus main protease.
The instrument used for the fluorescence intensity measurement was a Fluoraskan Ascent fluorometer (ThermoLabsystems, Helsinki, Finland), and the wavelengths of the excitation light and the emission light were 320nm and 405nm, respectively.
To a buffer solution (50mM Tris-HCl (pH 7.3), 1mM EDTA (with or without DTT)) was added SARS coronavirus main protease (final concentration 0.5. mu.M), DMSO lysate of compound (to make the final concentration: 500. mu.g/mL, substrate concentration 20. mu.M, after 10 minutes at 298K, fluorescently labeled substrate (MCA-AVLQSGFRL(DNP) L-NH2, 20 μ M final) was added rapidly, negative controls were set: the compound is not added to the reaction mixture, the other conditions are the same, the excitation wavelength and the emission wavelength are respectively 320nm and 405nm, the temperature is kept at 298K, the fluorescence reading is recorded every 10 seconds, 10 points are totally determined, the time is taken as an X axis, the fluorescence value is taken as a Y axis to plot to obtain an enzyme activity mechanical curve, the initial speed V of the reaction can be obtained by calculating the slope through the numerical values of the first two points on the graph, and the initial speed of the reaction of the negative control is defined as V.0The initial rate of reaction of the added compound is defined as ViTo calculate the residual activity of the host protease after addition of the corresponding compound (V)i/V0) The inhibition ratio of the corresponding compound is (1-V)i/V0). When the remaining activity of the host protease is less than 20% (or the inhibition rate is greater than 80%), the IC of the compound is further determined50The value is obtained.
3. IC of compound for inhibiting SARS coronavirus main proteinase activity50Determination of value
The compounds were dissolved in 95% DMSO to 50mmol/L solutions, respectively. 10. mu.L of the solution was diluted with 95% DMSO in a gradient of 2 times to obtain about 12 sample solutions. The residual activity of the main protease was tested at various chemical concentrations using the assay described above. The diluted compound concentrations were divided by the molecular weights of the corresponding compounds and the corresponding residual activity values were plotted on the X-axis against the base 10 logarithmic values and on the Y-axis against GraphPad Prism 5(GraphPad software Inc.) and the corresponding IC's were calculated50The value is obtained.
IC of Compounds 1 to 550The values were 37uM, 78uM, 55uM, 49uM and 29uM, respectively. Wherein the inhibition rate is 500. mu. mol.L－1Measured at the concentration of (c).
It should be clear that throughout the description, the purpose of describing a preferred embodiment of the invention is not to limit the invention to any one embodiment or specific set of features. Thus, it will be understood by those skilled in the art from this disclosure that various modifications and changes can be made to the specific embodiments described without departing from the scope of the invention. All such modifications and variations are intended to be included herein within the scope of the appended claims.
Claims (8)
1. A compound of formula I:
wherein,
R1is methyl, hydrogen or
R2Is selected fromGroup (d) of (a).
2. The compound of claim 1, which is:
3. the compound of claim 1, which is:
4. the compound of claim 1, which is:
5. the compound of claim 1, which is:
6. the compound of claim 1, which is:
7. use of a compound according to any one of claims 1 to 6 in the manufacture of a medicament for the treatment of severe acute respiratory syndrome.
8. Use of a compound according to any one of claims 1 to 6 in the manufacture of a medicament for the treatment of SARS virus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110446695.2A CN103183631B (en) | 2011-12-28 | 2011-12-28 | Isatin-5-sulfonic acid amide derivatives and the application in the medicine of preparation treatment severe acute respiratory syndrome thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110446695.2A CN103183631B (en) | 2011-12-28 | 2011-12-28 | Isatin-5-sulfonic acid amide derivatives and the application in the medicine of preparation treatment severe acute respiratory syndrome thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103183631A CN103183631A (en) | 2013-07-03 |
| CN103183631B true CN103183631B (en) | 2016-08-24 |
Family
ID=48675109
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110446695.2A Expired - Fee Related CN103183631B (en) | 2011-12-28 | 2011-12-28 | Isatin-5-sulfonic acid amide derivatives and the application in the medicine of preparation treatment severe acute respiratory syndrome thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103183631B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108299280A (en) * | 2018-01-19 | 2018-07-20 | 天津国际生物医药联合研究院 | Isatin -5- the sulfonamide inhibitors inhibited to mixed lineage leukemia key protein |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001022966A1 (en) * | 1999-09-30 | 2001-04-05 | Smithkline Beecham Corporation | Caspases and apoptosis |
| US6403792B1 (en) * | 1997-07-30 | 2002-06-11 | Smithkline Beecham Corporation | Sulfonyl isatin compounds and methods of blocking apoptosis therewith |
| CN102006865A (en) * | 2008-02-29 | 2011-04-06 | 遗传工程与生物技术中心 | Chemical compounds having antiviral activity against dengue virus and other flaviviruses |
| CN102171208A (en) * | 2008-09-05 | 2011-08-31 | 帝国创新有限公司 | Isatin derivatives for use as in vivo imaging agents |
-
2011
- 2011-12-28 CN CN201110446695.2A patent/CN103183631B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6403792B1 (en) * | 1997-07-30 | 2002-06-11 | Smithkline Beecham Corporation | Sulfonyl isatin compounds and methods of blocking apoptosis therewith |
| WO2001022966A1 (en) * | 1999-09-30 | 2001-04-05 | Smithkline Beecham Corporation | Caspases and apoptosis |
| CN102006865A (en) * | 2008-02-29 | 2011-04-06 | 遗传工程与生物技术中心 | Chemical compounds having antiviral activity against dengue virus and other flaviviruses |
| CN102171208A (en) * | 2008-09-05 | 2011-08-31 | 帝国创新有限公司 | Isatin derivatives for use as in vivo imaging agents |
Non-Patent Citations (7)
| Title |
|---|
| Docking and 3D-QSAR Studies on Isatin Sulfonamide Analogues as Caspase-3 Inhibitors;Qi Wang, 等;《J.Chem.Inf.Model.》;20090717;第49卷(第8期);第1965-1966页表格1 * |
| Isatin 1,2,3-triazoles as potent inhibitors against caspase-3;Yang Jiang, 等;《Bioorg. Med. Chem.》;20110131;第21卷(第6期);第1627页Scheme 1. * |
| Isatin Compounds as Noncovalent SARS Coronavirus 3C-like Protease Inhibitors;Lu Zhou, 等;《J. Med. Chem.》;20060516;第49卷(第12期);第3440-3443页 * |
| N-Benzylisatin Sulfonamide Analogues as Potent Caspase-3 Inhibitors: Synthesis, in Vitro Activity, and Molecular Modeling Studies;Wenhua Chu, 等;《J. Med. Chem.》;20051105;第48卷(第24期);第7638-7639页Scheme 1-Scheme 3 * |
| Structure–Activity Correlation for a Series of Isatin Derivatives, Inhibitors of Caspase 3;I.G.Tsygankova,等;《Russian Journal of General Chemistry》;20091231;第79卷(第3期);第489页化合物22-36、38-51 * |
| Synthesis and in Vitro Evaluation of Sulfonamide Isatin Michael Acceptors as Small Molecule Inhibitors of Caspase-6;Wenhua Chu, 等;《J. Med. Chem.》;20090330;第52卷(第8期);第2190页Scheme 1 * |
| 非肽类SARS冠状病毒3CL蛋白酶抑制剂的设计与活性表征;刘莹,等;《化学学报》;20070828;第65卷(第16期);第1707-1712页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103183631A (en) | 2013-07-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Steuer et al. | Synthesis and biological evaluation of α-ketoamides as inhibitors of the Dengue virus protease with antiviral activity in cell-culture | |
| TWI406856B (en) | Inhibitors of human immunodeficiency virus replication | |
| EP1481965B1 (en) | Aromatic amino acid derivatives and medicinal compositions | |
| US11820763B2 (en) | Bromophenol-pyrazoline compound and synthesis method and use thereof | |
| Li et al. | Discovering novel chemical inhibitors of human cyclophilin A: virtual screening, synthesis, and bioassay | |
| Pisani et al. | Fine molecular tuning at position 4 of 2H-chromen-2-one derivatives in the search of potent and selective monoamine oxidase B inhibitors | |
| CN111803501B (en) | Use of chiral chloroquine hydroxychloroquine for reducing cardiotoxicity | |
| CN112920136B (en) | Compound and medical application thereof in novel coronavirus pneumonia | |
| CN103159665B (en) | Isatin-5-amide inhibiting agent with inhibition effect against SARS coronavirus main protease | |
| Cui et al. | Design, synthesis, bioactivity, and DFT calculation of 2-thiazolyl-hydrazone derivatives as influenza neuraminidase inhibitors | |
| Zhang et al. | Design, synthesis and biological evaluation of N-(4-alkoxy-3-(1H-tetrazol-1-yl) phenyl) heterocyclic aromatic amide derivatives as xanthine oxidase inhibitors | |
| EP1636200A2 (en) | Inhibitors of papilloma virus | |
| JP2003509337A (en) | Substituted benzamide inhibitors of rhinovirus 3C protease | |
| CN103183631B (en) | Isatin-5-sulfonic acid amide derivatives and the application in the medicine of preparation treatment severe acute respiratory syndrome thereof | |
| Yan et al. | Structural modifications of CH (OH)-DAPYs as new HIV-1 non-nucleoside reverse transcriptase inhibitors | |
| CN100502868C (en) | 3CL protease inhibitor of non-peptide SARS coronavirus and use thereof | |
| CN115594655B (en) | Chromone oxime derivative and preparation method and application thereof | |
| Zhou et al. | Structure-based optimization of Toddacoumalone as highly potent and selective PDE4 inhibitors with anti-inflammatory effects | |
| EP2810942B1 (en) | Paroxetine derivative | |
| CN110759891B (en) | SET8 lysine methyltransferase inhibitor and intermediate, preparation method and application thereof | |
| CN109651119A (en) | A kind of piperazine Lun Tawei intermediate synthetic method | |
| CN103159666A (en) | Isatin derivative and purpose thereof | |
| Ahn et al. | Synthesis and evaluation of benzoquinolinone derivatives as sars-cov 3cl protease inhibitors | |
| CN103230393A (en) | Application of isatin derivatives in preparing anti-SARS medicines | |
| Ou et al. | Design, synthesis, and nematicidal activity of novel 1, 2, 4-oxadiazole derivatives containing amide fragments |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160824 Termination date: 20211228 |