CN103626705A - 1-(3-benzoylaminobenzyl)-1H-indazole-3-carboxamide compounds, and preparation method and antiviral use thereof - Google Patents

1-(3-benzoylaminobenzyl)-1H-indazole-3-carboxamide compounds, and preparation method and antiviral use thereof Download PDF

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CN103626705A
CN103626705A CN201210308339.9A CN201210308339A CN103626705A CN 103626705 A CN103626705 A CN 103626705A CN 201210308339 A CN201210308339 A CN 201210308339A CN 103626705 A CN103626705 A CN 103626705A
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compound
indazole
carbonyl
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CN103626705B (en
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李援朝
左建平
施晶晶
何珮岚
唐炜
童贤崑
冯春兰
王桂凤
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Shanghai Institute of Materia Medica of CAS
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
    • C07D231/54—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings condensed with carbocyclic rings or ring systems
    • C07D231/56—Benzopyrazoles; Hydrogenated benzopyrazoles

Abstract

The invention relates to virus inhibitors, and concretely relates to 1-(3-benzyl-benzoylaminobenzyl)-1H-indazole-3-carboxamide organic micro-molecular compounds having a structure represented by general formula 1 and used as non-nucleoside virus inhibitors, pharmaceutically acceptable salts or acceptable hydrates thereof, a preparation method thereof, and an application of the compounds in the preparation of drugs for treating viral diseases comprising hepatitis C, hepatitis B, influenzas, herpes, AIDS and the like, especially an application in the preparation of drugs for treating the hepatitis C. The compounds have the advantages of simple synthesis, easily available raw materials and low toxicity.

Description

1-(3-benzamido benzyl)-1H-indazole-3-Carbox amide and preparation method thereof and anti-viral uses
Technical field
The invention belongs to field of medicaments, relate to one antiviral inhibitor, particularly, relate to 1-(3-benzamido benzyl)-1H-indazole-3-benzamide type organic micromolecule compound that a class can be used as non-nucleoside antivirus inhibitor, and pharmacy acceptable salt or hydrate, its preparation method with and application, the particularly application in the medicine for the preparation for the treatment of hepatitis C in the medicine for the preparation of virus diseases such as treatment hepatitis C, hepatitis B, influenza, bleb, acquired immune deficiency syndrome (AIDS).
Background technology
The antiviral therapy medicine of approved is less at present, is mainly nucleoside medicine: ribavirin, acyclovir and ganciclovir etc.And chronically infected treatment is only confined to adopt IFN-α and ribavirin antiviral pathway to hepatitis C virus (HCV).Therefore, explore with hepatitis B virus resisting (HBV), the HCV medicine of exploitation high-efficiency low-toxicity and become current problem demanding prompt solution.
Non-nucleoside antiviral is because its potential particular mechanism of action may be avoided the generation of resistance, for frontier, new approaches have been opened up in antiviral research.Thereby exploitation high specificity, the novel non-nucleoside antiviral with clinical efficacy that toxic side effect is little are important research directions of new drug research.
Summary of the invention
An object of the present invention is to provide the 1-shown in general formula 1 (3-benzamido benzyl)-1H-indazole-3-Carbox amide and pharmacy acceptable salt or hydrate.
Another object of the present invention is to provide above-mentioned 1-(3-benzamido benzyl)-1H-indazole-3-preparation method of Carbox amide.
Another object of the present invention is to provide a kind of pharmaceutical composition, its comprise one or more treatment significant quantities according to the 1-shown in general formula 1 of the present invention (3-benzamido benzyl)-1H-indazole-3-Carbox amide and pharmacy acceptable salt or hydrate, optionally, pharmaceutically acceptable conventional auxiliary material.
A further object of the present invention is to provide the application in the medicine for the preparation for the treatment of virus infection, particularly hepatitis C of above-mentioned 1-(3-benzamido benzyl)-1H-indazole-3-Carbox amide and pharmacy acceptable salt or hydrate thereof.
According to an aspect of the present invention, the compound and pharmacy acceptable salt or the hydrate that provide a class to there is structure shown in following general formula 1:
Figure BDA00002060500300021
R wherein 1, R 2identical or different, be H, C independently of one another 1-C 10the alkyl of straight or branched, C 1-C 10the alkoxyl group of straight or branched, C 2-C 10the alkenyl of straight or branched, C 3-C 10cycloalkyl or C 5-C 20aryl; Be preferably H, C 1-C 6the alkyl of straight or branched, C 1-C 6the alkoxyl group of straight or branched, C 2-C 6the alkenyl of straight or branched, C 3-C 8cycloalkyl or C 5-C 12aryl; And most preferably be H, methyl, ethyl, propyl group, sec.-propyl, butyl, the tertiary butyl, methoxyl group, oxyethyl group, propoxy-, isopropoxy, butoxy, tert.-butoxy, vinyl, propenyl, butenyl, pentenyl, hexenyl, cyclopropane base, tetramethylene base, pentamethylene base, cyclohexyl, suberane base or phenyl;
Wherein, C 5-C 20aromatic base is preferably phenyl, thienyl, furyl, pyridyl, pyrimidyl, pyrryl, imidazolyl, oxazolyl, indazolyl, indyl, quinolyl, naphthyl, isoquinolyl, benzofuryl, benzothienyl or benzothiazolyl;
R 3, R 4, R 5, R 6, R 7identical or different, and can be H, halogen, C independently of one another 1-C 10the alkyl of straight or branched, C 1-C 10the alkoxyl group of straight or branched, C 2-C 10the alkenyl of straight or branched, C 3-C 10cycloalkyl, CF 3,-NO 2,-CN, hydroxyl, C 1-C 10alkyl-carbonyl or C 1-C 10alkoxy carbonyl; More preferably H, F, Br, Cl, I, C 1-C 6the alkyl of straight or branched, C 1-C 6the alkoxyl group of straight or branched, C 2-C 6the alkenyl of straight or branched, C 3-C 8cycloalkyl, CF 3, x-NO 2,-CN, hydroxyl, C 1-C 6alkyl-carbonyl or C 1-C 6alkoxy carbonyl; More preferably H, F, Br, Cl, I, methyl, ethyl, propyl group, sec.-propyl, butyl, the tertiary butyl, methoxyl group, oxyethyl group, propoxy-, isopropoxy, butoxy, tert.-butoxy, vinyl, propenyl, butenyl, pentenyl, hexenyl, cyclopropane base, tetramethylene base, pentamethylene base, cyclohexyl, suberane base, CF 3,-NO 2,-CN, hydroxyl, methyl carbonyl, ethyl carbonyl, propyl group carbonyl, sec.-propyl carbonyl, butyl carbonyl, tertiary butyl carbonyl, methoxycarbonyl, ethoxy carbonyl, propoxycarbonyl, isopropoxy carbonyl, butoxy carbonyl or tert-butoxycarbonyl.
More specifically, the described compound being represented by general formula 1 can be any one in following compound:
Table 1
Figure BDA00002060500300022
Figure BDA00002060500300031
Figure BDA00002060500300041
Figure BDA00002060500300051
Figure BDA00002060500300061
Figure BDA00002060500300071
Figure BDA00002060500300081
Term in the present invention " pharmacy acceptable salt " refers to and the mineral acids such as phosphoric acid, sulfuric acid, hydrochloric acid, or the organic acid such as acetic acid, tartrate, citric acid, oxysuccinic acid, or the salt that forms of the acidic amino acid such as aspartic acid, L-glutamic acid, or after becoming ester or acid amides with above-mentioned acid again with the salt of mineral alkali formation, as sodium, potassium, calcium, aluminium salt and ammonium salt.
The synthetic method of the 1-shown in general formula 1 of the present invention (3-benzamido benzyl)-1H-indazole-3-Carbox amide comprises the steps:
Figure BDA00002060500300082
Reaction stream formula 1
A) take 3-formic acid-1H-indazole is initiator, and under the effect of thionyl chloride or oxalyl chloride, reaction generates after acyl chlorides and amine HNR again 1r 2there is amidate action and obtain the compound shown in general formula 2;
B) there is the compound shown in N-1 position substitution reaction generation general formula 3 with 1-(bromomethyl)-3-oil of mirbane in the compound shown in general formula 2 under the effect of alkali;
C) then the compound shown in general formula 3 is reduced to the compound shown in general formula 4 under the effect of reductive agent;
D) compound shown in general formula 4 under amide condensed dose of effect with
Figure BDA00002060500300091
or with
Figure BDA00002060500300092
there is amidate action and generate the compound shown in general formula 1.
Wherein, R 1, R 2, R 3, R 4, R 5, R 6and R 7definition identical with definition above.
Step b wherein) alkali in can be sodium carbonate, salt of wormwood, sodium hydroxide, potassium hydroxide, sodium methylate, sodium ethylate, sodium ethylate, potassium ethylate, sodium hydride or potassium hydride KH etc.
Step c wherein) reductive agent in can be tindichloride, iron powder or zinc powder etc.
Steps d wherein) amide condensed dose in can be carbonyl dimidazoles (CDI), dicyclohexylcarbodiimide (DCC), DIC (DIC), 1-(3-dimethylamino-propyl)-3-ethyl carbodiimide (EDCI), 2-(7-azo benzotriazole)-N, N, N', N'-tetramethyl-urea phosphofluoric acid ester (HATU), benzotriazole-N, N, N', N'-tetramethyl-urea hexafluorophosphate (HBTU), 6-Chloro-Benzotriazole-1, 1, 3, 3-tetramethyl-urea hexafluorophosphate (HCTU), O-benzotriazole-N, N, N', N'-tetramethyl-urea Tetrafluoroboric acid ester (TBTU), 6-Chloro-Benzotriazole-1, 1, 3, 3-tetramethyl-urea Tetrafluoroboric acid ester (TCTU), 2-succinimido-1, 1, 3, 3-tetramethyl-urea Tetrafluoroboric acid ester (TSTU) or 2-(5-norbornylene-2, 3-dicarboximide base)-1, 1, 3, 3-tetramethyl-urea tetrafluoroborate (TNTU) etc.
Another aspect of the present invention provides a kind of pharmaceutical composition for the treatment of virus infection, and it comprises being selected from according to one or more in above-mentioned 1-of the present invention (3-benzamido benzyl)-1H-indazole-3-Carbox amide, its pharmacy acceptable salt and hydrate for the treatment of significant quantity.
One side more of the present invention provides the application in the medicine for the preparation for the treatment of virus infection, particularly hepatitis C of above-mentioned 1-of the present invention (3-benzamido benzyl)-1H-indazole-3-Carbox amide, its pharmacy acceptable salt or hydrate.
Another aspect of the present invention provides a kind of virus infection for the treatment of, the method of hepatitis C particularly, it comprises to patient's administration of this treatment of needs and being selected from according to one or more in above-mentioned 1-of the present invention (3-benzamido benzyl)-1H-indazole-3-Carbox amide, its pharmacy acceptable salt and hydrate.
Embodiment
Below in conjunction with embodiment, the present invention is further elaborated, but these embodiment are never any limitation of the invention, scope of the present invention is determined by claim.
Preparation Example
Instrument and main experiment material are as follows:
BrukerAM-400 type and Varian Mercury plus-400 type nuclear magnetic resonance analyser, Agilent 6110 type mass spectrographs, and 200-300 order column chromatography silica gel (Haiyang Chemical Plant, Qingdao), HSGF254TLC plate (Yantai City chemical research institute).
Preparation Example 1:
The preparation of compound 33:1H-indazole-3-methane amide
Figure BDA00002060500300101
Under argon shield; mixing solutions reflux 3h by raw material 3-formic acid-1H-indazole (5g, 30.9mmol) with 20ml thionyl chloride, steams excessive thionyl chloride and obtains yellow solid; the yellow solid of gained is dissolved in anhydrous 20ml tetrahydrofuran (THF); slowly drip strong aqua 5ml, in process, have solid to separate out, after stirring at room 30min, filter; washing; be dried to obtain compound 33, be white solid (3.8g, productive rate 76%). 1H NMR(300MHz,DMSO)δ:13.49(s,1H),8.15(d,J=8.2Hz,1H),7.69(s,1H),7.58(d,J=8.5Hz,1H),7.38(t,J=7.7Hz,1H),7.30(s,1H),7.21(t,J=7.4Hz,1H).MS(ESI)m/z:184(M+Na)。
Preparation Example 2:
The preparation of compound 34:1-(3-nitrobenzyl)-1H-indazole-3-methane amide
Figure BDA00002060500300102
Compound 33 (2.5g, 15.5mmol) is dissolved in 20ml DMF, in 0 ℃, adds nitro benzyl bromide (3.4g, 15.5mmol) and a K 2cO 3(4.3mg, 30.1mmol), stirring at room adds 50ml water after 4 hours, separate out white solid, filters, and water, ether washing, be dried to obtain compound 34 successively, is white solid (4.4g, productive rate: 95%). 1H NMR(300MHz,DMSO)δ:8.16(m,3H),7.83(d,J=8.7Hz,1H),7.74(s,1H),7.63(m,2H),7.44(m,2H),7.27(t,J=7.5Hz,1H),5.89(s,2H).MS(ESI)m/z:319(M+Na)。
Preparation Example 3:
The preparation of compound 35:1-(3-aminobenzyl)-1H-indazole-3-methane amide
Figure BDA00002060500300111
By compound 34 (4.0g, 13.5mmol) be dissolved in 20ml tetrahydrofuran (THF), under room temperature, add two hydrated stannous chloride 15.2g, 67.6mmol) room temperature reaction 12 hours, add 10% aqueous sodium hydroxide solution to adjust pH to alkalescence, ethyl acetate extraction, saturated sodium hydroxide washing, dry, concentrated, column chromatographic isolation and purification obtains compound 35, is white solid (2.8g, productive rate 78%). 1H NMR(300MHz,DMSO)δ:8.17(d,J=8.1Hz,1H),7.67(d,J=8.6Hz,2H),7.40(m,2H),7.23(t,J=7.5Hz,1H),6.93(t,J=7.5Hz,1H),6.40(m,2H),6.32(s,1H),5.55(s,2H),5.10(s,2H). 13C NMR(101MHz,DMSO)δ:164.38,149.41,141.14,137.87,137.84,129.57,127.05,123.01,122.79,122.47,115.07,113.76,112.68,111.01,53.19.MS(ESI)m/z:289(M+Na)。
Preparation Example 4:
Compound 1:1-(3-(benzamido) benzyl)-1H-indazole-3-methane amide
Figure BDA00002060500300112
Method one: by compound 35 (80mg, 0.3mmol) with triethylamine (86 μ l, 0.6mmol) be dissolved in 2ml methylene dichloride, slowly add Benzoyl chloride (86 μ l, 0.33mmol), stirring at room 30min, pressure reducing and steaming solvent, column chromatographic isolation and purification obtains compound 1, is white solid (102mg, productive rate 92%).
Method two: by compound 35 (80mg, 0.3mmol), phenylformic acid (40mg, 0.33mmol), diisopropylethylamine (0.157ml, 0.9mmol) is dissolved in 2ml N, in dinethylformamide, add HBTU (151mg, 0.39mmol), stirring at room 2h, ethyl acetate extracts, water, saturated common salt water washing, dry, the concentrated crude product that to obtain, and column chromatographic isolation and purification obtains compound 1, for white solid (93mg, productive rate 86%). 1H NMR(300MHz,DMSO)δ:10.22(s,1H),8.19(d,J=8.1Hz,1H),7.88(d,J=7.0Hz,2H),7.70m,3H),7.48(m,5H),7.27(m,2H),6.98(d,J=7.7Hz,1H),5.73(s,2H). 13C NMR(100MHz,DMSO)δ:165.66,163.87,140.68,139.45,137.59,137.34,134.78,131.59,128.93,128.35×2,127.63×2,126.75,122.62,122.51,122.44,122.03,119.74,119.05,110.46,52.41.MS(ESI)m/z:393(M+Na).M.P.:206-208℃。
Preparation Example 5:
Compound 2:1-(3-(2-fluorobenzoyl is amino) benzyl)-1H-indazole-3-methane amide
Except using 2-fluorobenzoyl chloride to replace, Benzoyl chloride, preparing compound 2 in the mode identical with Preparation Example 4, be white solid, yield is 80%. 1H NMR(300MHz,DMSO)δ:10.39(s,1H),8.18(d,J=8.1Hz,1H),7.62(m,6H),7.42(m,2H),7.27(m,4H),6.99(d,J=7.5Hz,1H),5.73(s,2H). 13CNMR(100MHz,DMSO)δ:164.32,163.31,159.31(d,J=248.8Hz),141.21,139.66,138.17,138.03,132.95(d,J=8.3Hz),130.30(d,J=2.7Hz),129.56,127.23,125.40(d,J=15.4Hz),124.99(d,J=3.4Hz),123.22,123.01,122.90,122.54,119.56,118.87,116.57(d,J=21.9Hz),110.91,52.83.MS(ESI)m/z:411(M+Na)。
Preparation Example 6:
Compound 3:1-(3-(3-fluorobenzoyl is amino) benzyl)-1H-indazole-3-methane amide
Except using 3-fluorobenzoyl chloride to replace, Benzoyl chloride, preparing compound 3 in the mode identical with Preparation Example 4, be white solid, yield is 82%. 1H NMR(300MHz,DMSO)δ:10.28(s,1H),8.19(d,J=8.1Hz,1H),7.70(m,6H),7.54(m,1H),7.42(m,3H),7.28(m,2H),7.01(d,J=7.5Hz,1H),5.74(s,2H). 13C NMR(100MHz,DMSO)δ:164.70(d,J=2.4Hz),164.32,162.37(d,J=244.5Hz),141.21,139.70,138.18,137.87,137.61(d,J=6.7Hz),131.01(d,J=8.0Hz),129.44,127.22,124.35(d,J=2.6Hz),123.37,123.04,122.89,122.55,120.36,119.66,118.94(d,J=21.3Hz),114.96(d,J=22.9Hz),110.92,52.91.MS(ESI)m/z:411(M+Na)。
Preparation Example 7:
Compound 4:1-(3-(4-fluorobenzoyl is amino) benzyl)-1H-indazole-3-methane amide
Except using 4-fluorobenzoyl chloride to replace, Benzoyl chloride, preparing compound 4 in the mode identical with Preparation Example 4, be white solid, yield is 77%. 1H NMR(300MHz,DMSO)δ:10.24(s,1H),8.19(d,J=8.1Hz,1H),7.96(m,2H),7.69(m,4H),7.33(m,6H),6.99(d,J=7.8Hz,1H),5.73(s,2H). 13CNMR(100MHz,DMSO)δ:165.00,164.55(d,J=249.0Hz),164.32,141.21,139.88,138.16,137.83,131.75(d,J=2.9Hz),130.88(d,J=9.1Hz)×2,129.40,127.21,123.19,123.04,122.89,122.55,120.30,119.61,115.75(d,J=21.8Hz)×2,110.93,52.92.MS(ESI)m/z:411(M+Na).M.P.:208-210℃。
Preparation Example 8:
Compound 5:1-(3-(2-methoxybenzoyl is amino) benzyl)-1H-indazole-3-methane amide
Except using 2-methoxy benzoyl chloride to replace, Benzoyl chloride, preparing compound 5 in the mode identical with Preparation Example 4, be white solid, yield is 82%. 1H NMR(300MHz,DMSO)δ:10.10(s,1H),8.18(d,J=8.1Hz,1H),7.67(m,4H),7.53(d,J=7.2Hz,1H),7.44(m,3H),7.25(m,2H),7.13(d,J=8.4Hz,1H),7.01(t,J=7.4Hz,1H),6.93(dd,J=7.6,0.5Hz,1H),5.72(s,2H),3.83(s,3H). 13C NMR(100MHz,DMSO)δ:165.10,164.30,156.90,141.21,139.87,138.16,137.93,132.42,130.00,129.48,127.19,125.54,123.03,122.87,122.81,122.53,120.92,119.51,118.95,112.47,110.94,56.34,52.88.MS(ESI)m/z:423(M+Na)。
Preparation Example 9:
Compound 6:1-(3-(3-methoxybenzoyl is amino) benzyl)-1H-indazole-3-methane amide
Except using 3-methoxy benzoyl chloride to replace, Benzoyl chloride, preparing compound 6 in the mode identical with Preparation Example 4, be white solid, yield is 72%. 1H NMR(300MHz,DMSO)δ:10.19(s,1H),8.19(d,J=8.1Hz,1H),7.70(m,4H),7.35(m,7H),7.12(m,1H),6.99(d,J=7.5Hz,1H),5.73(s,2H),3.80(s,3H). 13C NMR(100MHz,DMSO)δ:165.81,164.31,159.65,141.21,139.91,138.17,137.80,136.73,129.98,129.38,127.20,123.16,123.04,122.88,122.55,120.34×2,119.70,117.80,113.43,110.94,55.83,52.94.MS(ESI)m/z:423(M+Na)。
Preparation Example 10:
Compound 7:1-(3-(4-methoxybenzoyl is amino) benzyl)-1H-indazole-3-methane amide
Except using 4-methoxy benzoyl chloride to replace, Benzoyl chloride, preparing compound 7 in the mode identical with Preparation Example 4, be white solid, yield is 77%. 1H NMR(300MHz,DMSO)δ:10.07(s,1H),8.19(d,J=8.1Hz,1H),7.89(d,J=8.1Hz,2H),7.70(m,4H),7.42(m,2H),7.26(m,2H),7.01(d,J=8.7Hz,2H),6.96(d,J=7.5Hz,1H),5.72(s,2H),3.80(s,3H). 13C NMR(100MHz,DMSO)δ:165.52,164.39,162.41,141.17,140.10,138.06,137.72,130.08×2,129.36,127.29,127.24,123.01×2,122.93,122.51,120.28,119.61,114.06×2,110.92,55.90,52.97.MS(ESI)m/z:423(M+Na).M.P.:208-210℃。
Preparation Example 11:
Compound 8:1-(3-(2-nitrobenzoyl amido) benzyl)-1H-indazole-3-methane amide
Except using 2-nitrobenzoyl chloride to replace, Benzoyl chloride, preparing compound 8 in the mode identical with Preparation Example 4, be white solid, yield is 70%. 1H NMR(300MHz,DMSO)δ:10.63(s,1H),8.18(d,J=8.2Hz,1H),8.11(d,J=8.2Hz,1H),7.82(dd,J=6.9,8.1Hz,1H),7.68(m,5H),7.35(m,5H),6.99(d,J=7.8Hz,1H),5.74(s,2H). 13C NMR(100MHz,DMSO)δ:164.63,164.29,146.85,141.24,139.66,138.21,138.17,134.56,133.07,131.43,129.74,129.65,127.24,124.69,123.24,123.00,122.90,122.55,119.41,118.65,110.91,52.76.MS(ESI)m/z:438(M+Na)。
Preparation Example 12:
Compound 9:1-(3-(3-nitrobenzoyl amido) benzyl)-1H-indazole-3-methane amide
Except using 3-nitrobenzoyl chloride to replace, Benzoyl chloride, preparing compound 9 in the mode identical with Preparation Example 4, be white solid, yield is 65%. 1H NMR(300MHz,DMSO)δ:10.56(s,1H),8.71(t,J=1.8Hz,1H),8.40(ddd,J=8.2,1.8,0.9Hz,1H),8.33(d,J=7.8Hz,1H),8.19(d,J=8.1Hz,1H),7.73(m,5H),7.34(m,4H),7.04(d,J=7.5Hz,1H),5.75(s,2H). 13C NMR(100MHz,DMSO)δ:164.32,163.93,148.21,141.21,139.49,138.19,137.95,136.68,134.65,130.63,129.51,127.24,126.62,123.63,123.03,122.89×2,122.55,120.50,119.75,110.92,52.88.MS(ESI)m/z:438(M+Na)。
Preparation Example 13:
Compound 10:1-(3-(4-nitrobenzoyl amido) benzyl)-1H-indazole-3-methane amide
Except using 4-nitrobenzoyl chloride to replace, Benzoyl chloride, preparing compound 10 in the mode identical with Preparation Example 4, be white solid, yield is 68%. 1H NMR(300MHz,DMSO)δ:10.54(s,1H),8.32(d,J=8.4Hz,2H),8.19(d,J=8.1Hz,1H),8.11(d,J=8.8Hz,2H),7.72(m,3H),7.61(s,1H),7.42(m,2H),7.33(t,J=7.8Hz,1H),7.25(t,J=7.5Hz,1H),7.04(d,J=7.5Hz,1H),5.75(s,2H). 13C NMR(100MHz,DMSO)δ:164.47,164.31,149.64,141.22,141.00,139.53,138.20,137.98,129.70×2,129.52,127.23,123.96×2,123.63,123.04,122.90,122.56,120.36,119.61,110.92,52.86.MS(ESI)m/z:438(M+Na).M.P.:209-210℃。
Preparation Example 14:
Compound 11:1-(3-(2-toluyl is amino) benzyl)-1H-indazole-3-methane amide
Except using 2-methyl benzoyl chloride to replace, Benzoyl chloride, preparing compound 11 in the mode identical with Preparation Example 4, be white solid, yield is 86%. 1H NMR(300MHz,DMSO)δ:10.27(s,1H),8.18(d,J=8.1Hz,1H),7.68(m,4H),7.33(m,8H),6.95(d,J=7.5Hz,1H),5.72(s,2H),2.32(s,3H). 13C NMR(100MHz,DMSO)δ:167.90,163.84,140.71,139.60,137.63,137.47,137.08,135.14,130.47,129.61,128.99,127.16,126.72,125.59,122.51,122.41,122.35,122.04,118.91,118.27,110.47,52.35,19.28.MS(ESI)m/z:407(M+Na)。
Preparation Example 15:
Compound 12:1-(3-(3-toluyl is amino) benzyl)-1H-indazole-3-methane amide
Except using 3-methyl benzoyl chloride to replace, Benzoyl chloride, preparing compound 12 in the mode identical with Preparation Example 4, be white solid, yield is 85%. 1H NMR(300MHz,DMSO)δ:10.18(s,1H),8.19(d,J=8.1Hz,1H),7.69(m,6H),7.34(m,6H),6.98(d,J=7.2Hz,1H),5.73(s,2H),2.36(s,3H). 13C NMR(100MHz,DMSO)δ:165.73,163.85,140.71,139.53,137.65×2,137.34,134.83,132.15,128.90,128.25,128.12,126.73,124.82,122.54×2,122.41,122.06,119.70,119.00,110.48,52.43,20.95.MS(ESI)m/z:407(M+Na)。
Preparation Example 16:
Compound 13:1-(3-(4-toluyl is amino) benzyl)-1H-indazole-3-methane amide
Except using 4-methyl benzoyl chloride to replace, Benzoyl chloride, preparing compound 13 in the mode identical with Preparation Example 4, be white solid, yield is 83%. 1H NMR(300MHz,DMSO)δ:10.14(s,1H),8.19(d,J=8.1Hz,1H),7.81(d,J=8.1Hz,2H),7.70(m,4H),7.42(m,2H),7.26(m,4H),6.97(d,J=7.5Hz,1H),5.72(s,2H),2.35(s,3H). 13C NMR(100MHz,DMSO)δ:165.43,163.86,141.61,140.70,139.56,137.63,137.31,131.91,128.87×3,127.69×2,126.73,122.53,122.49,122.41,122.06,119.73,119.05,110.47,52.45,21.01.MS(ESI)m/z:407(M+Na).M.P.:207-208℃。
Preparation Example 17:
Compound 14:1-(3-(3,4-difluorobenzene formamido group) benzyl)-1H-indazole-3-methane amide
Except using 3,4-difluoro benzoyl chloride to replace, outside Benzoyl chloride, preparing compound 14 in the mode identical with Preparation Example 4, be white solid, yield is 87%. 1H NMR(300MHz,DMSO)δ：10.28(s,1H),8.19(d,J=8.0Hz,1H),7.96(m,1H),7.79(m,1H),7.63(m,5H),7.42(m,2H),7.31(t,J=7.8Hz,1H),7.25(t,J=7.5Hz,1H),7.02(d,J=7.6Hz,1H),5.73(s,2H). 13C NMR(100MHz,dmso)δ:163.86,163.31,150.31(dd,J=494.7,13.0Hz),150.29(dd,J=12.8,2.5Hz),140.71,139.13,137.68,137.44,132.12(t,J=40.2Hz),128.99,126.75,125.29(dd,J=7.3,3.3Hz),122.95,122.54,122.43,122.08,119.86,119.10,117.39(dd,J=40.7,18.0Hz)×2,110.45,52.39.MS(ESI)m/z:429(M+Na)。
Preparation Example 18:
Compound 15:1-(3-(2,4 difluorobenzene formamido group) benzyl)-1H-indazole-3-methane amide
Except using 2,4 difluorobenzene formyl chloride to replace, Benzoyl chloride, preparing compound 15 in the mode identical with Preparation Example 4, be white solid, yield is 84%.1H NMR(300MHz,DMSO)δ：10.39(s,1H),8.20(d,J=8.1Hz,1H),7.68(m,4H),7.54(s,1H),7.30(m,6H),7.01(d,J=7.6Hz,1H),5.74(s,2H).13C NMR(100MHz,dmso)δ:163.84,163.37(dd,J=248.8,12.2Hz),161.96,159.47(dd,J=250.4,12.8Hz),140.71,139.10,137.69,137.62,131.62(dd,J=10.3,4.2Hz),131.55,129.11,126.75,122.80,122.53,122.42,122.07,121.59(dd,J=15.2,3.4Hz),119.06,118.30,111.80(dd,J=21.6,3.3Hz),110.44,104.60(t,J=26.2Hz),52.30.MS(ESI)m/z:429(M+Na)。
Preparation Example 19:
Compound 16:1-(3-(2,4,6-trifluoromethyl benzonitrile amido) benzyl)-1H-indazole-3-methane amide
Except using 2,4,6-trifluorobenzoyl chloride to replace, outside Benzoyl chloride, preparing compound 16 in the mode identical with Preparation Example 4, be white solid, yield is 88%. 1H NMR(300MHz,DMSO)δ:10.76(s,1H),8.20(d,J=7.8Hz,1H),7.68(m,3H),7.35(m,7H),7.02(d,J=7.5Hz,1H),5.75(s,2H). 13C NMR(100MHz,dmso)δ:163.83,161.41(t,J=15.8Hz),160.50(dd,J=15.8,10.8Hz),158.02(dd,J=15.8,10.8Hz),157.38,140.74,138.66,137.89,137.74,129.34,126.79,123.12,122.51,122.45,122.08,118.70,117.91,112.34(td,J=23.0,5.0Hz),110.42,101.21(t,J=28.2Hz)×2,52.17.MS(ESI)m/z:447(M+Na)。
Preparation Example 20:
Compound 17:1-(3-(4-trifluoromethyl benzamido) benzyl)-1H-indazole-3-methane amide
Except using 4-trifluoromethyl benzoyl chloride to replace, Benzoyl chloride, preparing compound 17 in the mode identical with Preparation Example 4, be white solid, yield is 82%. 1H NMR(300MHz,DMSO)δ:10.45(s,1H),8.20(d,J=8.1Hz,1H),8.08(d,J=8.1Hz,2H),7.86(d,J=8.1Hz,2H),7.70(m,4H),7.42(t,J=7.5Hz,2H),7.32(t,J=7.8Hz,1H),7.25(t,J=7.5Hz,1H),7.04(d,J=7.5Hz,1H),5.75(s,2H). 13C NMR(101MHz,dmso)δ:164.48,163.86,140.72,139.17,138.64,137.69,137.49,131.36(q,J=31.9Hz),129.01,128.61×2,126.74,126.62(q,J=269.9Hz),125.33(d,J=3.6Hz)×2,125.26,122.97,122.55,122.42,122.08,119.80,119.04,110.45,52.37.MS(ESI)m/z:461(M+Na)。
Preparation Example 21:
Compound 18:1-(3-(the fluoro-4-trifluoromethyl of 3-benzamido) benzyl)-1H-indazole-3-methane amide
Except using 3-fluorine 4-trifluoromethyl benzoyl chloride to replace, Benzoyl chloride, preparing compound 18 in the mode identical with Preparation Example 4, be white solid, yield is 79%. 1H NMR(300MHz,DMSO)δ:10.47(s,1H),8.20(d,J=7.8Hz,1H),7.93(m,3H),7.68(m,4H),7.42(m,2H),7.33(t,J=7.8Hz,1H),7.25(t,J=7.5Hz,1H),7.06(d,J=7.5Hz,1H),5.75(s,2H). 13C NMR(100MHz,dmso)δ:163.88,163.10,158.63(d,J=255.5Hz),141.37(d,J=7.4Hz),140.73,138.91,137.71,137.56,129.09,127.69(d,J=4.5Hz),126.79,124.35(d,J=3.3Hz),,123.25,122.56,122.47,122.37(q,J=272.2Hz),122.10,119.89,119.09,118.74(m),116.41(d,J=22.1Hz),110.47,52.36.MS(ESI)m/z:479(M+Na)。
Preparation Example 22:
Compound 19:1-(3-(the fluoro-4-trifluoromethyl of 2-benzamido) benzyl)-1H-indazole-3-methane amide
Except using 2-fluorine 4-trifluoromethyl benzoyl chloride to replace, Benzoyl chloride, preparing compound 19 in the mode identical with Preparation Example 4, be white solid, yield is 81%. 1H NMR(300MHz,DMSO)δ:10.59(s,1H),8.20(d,J=8.2Hz,1H),7.82(d,J=8.3Hz,2H),7.70(dd,J=14.9,8.7Hz,4H),7.52(s,1H),7.44(d,J=8.2Hz,2H),7.33(t,J=7.9Hz,1H),7.25(t,J=7.5Hz,1H),7.04(d,J=7.6Hz,1H),5.75(s,2H). 13C NMR(100MHz,dmso)δ:163.87,161.65,158.56(d,J=251.1Hz),140.74,138.89,137.76,137.73,132.28(m),131.14(d,J=3.0Hz),129.23,128.90(d,J=16.0Hz),126.79,123.08,123.05(q,J=271.2Hz),122.55,122.46,122.10,121.53(t,J=3.5Hz),119.04,118.24,113.86(m),110.45,52.27.MS(ESI)m/z:479(M+Na)。
Preparation Example 23:
Compound 20:1-(3-(2,4-dimethyl benzene formamido group) benzyl)-1H-indazole-3-methane amide
Except using 2,3-dimethyl benzoyl chloride to replace, outside Benzoyl chloride, preparing compound 20 in the mode identical with Preparation Example 4, be white solid, yield is 82%. 1H NMR(300MHz,DMSO)δ：10.26(s,1H),8.19(d,J=8.1Hz,1H),7.68(m,4H),7.42(m,2H),7.20(m,5H),6.94(d,J=7.9Hz,1H),5.72(s,2H),2.25(s,3H),2.19(s,3H). 13C NMR(100MHz,dmso)δ:168.53,163.85,140.72,139.61,137.93,137.63,137.49,137.14,133.18,130.61,129.00,126.73,125.33,124.59,122.51,122.41,122.31,122.04,118.83,118.20,110.47,52.34,19.81,16.05.MS(ESI)m/z:421(M+Na)。
Preparation Example 24:
Compound 21:1-(3-(3,4-dimethyl benzene formamido group) benzyl)-1H-indazole-3-methane amide
Except using 3,4-dimethyl benzoyl chloride to replace, outside Benzoyl chloride, preparing compound 21 in the mode identical with Preparation Example 4, be white solid, yield is 89%. 1H NMR(300MHz,DMSO)δ:10.10(s,1H),8.20(d,J=8.0Hz,1H),7.68(m,6H),7.42(m,2H),7.27(m,3H),6.97(d,J=7.3Hz,1H),5.72(s,2H),2.26(s,6H). 13C NMR(100MHz,dmso)δ:165.58,163.87,140.71,140.34,139.63,137.64,137.30,136.28,132.27,129.35,128.88,128.64,126.73,125.19,122.55,122.42×2,122.07,119.71,119.02,110.48,52.47,19.41×2.MS(ESI)m/z:421(M+Na)。
Preparation Example 25:
Compound 22:1-(3-(2-methoxyl group-4-toluyl is amino) benzyl)-1H-indazole-3-methane amide
Except using 2-methoxyl group 4-methyl benzoyl chloride to replace, Benzoyl chloride, preparing compound 22 in the mode identical with Preparation Example 4, be white solid, yield is 85%. 1H NMR(300MHz,DMSO)δ:10.00(s,1H),8.19(d,J=8.1Hz,1H),7.72(m,3H),7.61(d,J=8.1Hz,1H),7.52(d,J=7.5Hz,1H),7.42(m,2H),7.25(td,J=7.5,2.5Hz,2H),6.97(s,1H),6.92(d,J=7.8Hz,1H),6.84(d,J=7.8Hz,1H),5.72(s,2H),3.85(s,3H),2.33(s,3H). 13C NMR(100MHz,dmso)δ:164.33,163.85,156.52,142.56,140.69,139.34,137.63,137.46,129.85,129.01,126.72,122.53,122.41,122.27,122.05,121.55,121.15,119.08,118.52,112.53,110.48,55.84,52.38,21.30.MS(ESI)m/z:437(M+Na)。
Preparation Example 26:
Compound 23:1-(3-(3-methoxyl group-4-nitrobenzoyl amido) benzyl)-1H-indazole-3-methane amide
Except using 3-methoxyl group 4-nitrobenzoyl chloride to replace, Benzoyl chloride, preparing compound 23 in the mode identical with Preparation Example 4, be white solid, yield is 86%. 1H NMR(300MHz,DMSO)δ:10.44(s,1H),8.20(d,J=7.8Hz,1H),7.97(d,J=8.4Hz,1H),7.66(m,5H),7.42(m,2H),7.34(t,J=7.8Hz,1H),7.25(t,J=7.5Hz,1H),7.07(d,J=7.5Hz,1H),5.75(s,2H),3.98(s,3H). 13C NMR(100MHz,dmso)δ:163.95,163.87,151.55,140.87,140.71,139.95,138.99,137.69,137.52,129.06,126.77,124.89,123.22,122.55,122.45,122.09,120.03,119.72,119.21,113.54,110.45,56.92,52.38.MS(ESI)m/z:468(M+Na)。
Preparation Example 27:
Compound 24:1-(3-(3,4,5-Three methyl Benzene formamido group) benzyl)-1H-indazole-3-methane amide
Except using 3,4,5-trimethoxy-benzoyl chloride to replace, outside Benzoyl chloride, preparing compound 24 in the mode identical with Preparation Example 4, be white solid, yield is 85%. 1H NMR(300MHz,DMSO)δ:10.11(s,1H),8.19(d,J=8.1Hz,1H),7.67(m,4H),7.42(m,2H),7.28(m,2H),7.21(s,2H),7.01(d,J=7.5Hz,1H),5.73(s,2H),3.83(s,6H),3.70(s,3H). 13C NMR(100MHz,dmso)δ:164.99,163.85,152.57×2,140.69,140.27,139.35,137.65,137.34,129.91,128.92,126.74,122.77,122.54,122.43,122.06,120.12,119.36,110.47,105.28×2,60.12,56.09×2,52.44.MS(ESI)m/z:483(M+Na)。
Preparation Example 28:
Compound 25:1-(3-(4-acetylbenzene formamido group) benzyl)-1H-indazole-3-methane amide
Except using 4-acetylbenzene formyl chloride to replace, Benzoyl chloride, preparing compound 25 in the mode identical with Preparation Example 4, yield is 89%. 1H NMR(300MHz,DMSO)δ:10.39(s,1H),8.20(d,J=8.0Hz,1H),8.03(m,4H),7.70(m,4H),7.42(m,2H),7.32(t,J=7.8Hz,1H),7.25(t,J=7.5Hz,1H),7.03(d,J=7.5Hz,1H),5.74(s,2H),2.61(s,3H). 13C NMR(100MHz,dmso)δ:197.68,164.84,163.85,140.70,139.25,138.81,138.60,137.67,137.43,128.98,128.12×2,128.02×2,126.73,122.89,122.54,122.41,122.07,119.81,119.07,110.44,52.39,26.98.MS(ESI)m/z:435(M+Na)。
Preparation Example 29:
Compound 26:1-(3-(4-methyl-formiate benzamido) benzyl)-1H-indazole-3-methane amide
Except using 4-methoxycarbonyl Benzoyl chloride to replace, Benzoyl chloride, preparing compound 26 in the mode identical with Preparation Example 4, be white solid, yield is 91%. 1H NMR(300MHz,DMSO)δ:10.41(s,1H),8.19(d,J=8.1Hz,1H),8.02(m,3H),7.69(m,4H),7.42(m,2H),7.32(t,J=7.8Hz,1H),7.25(t,J=7.5Hz,1H),7.02(d,J=7.5Hz,1H),5.74(s,2H),3.87(s,3H). 13C NMR(100MHz,dmso)δ:165.66,164.78,163.84,140.71,139.22,138.91,137.67,137.45,132.01,129.13×2,128.98,128.08×2,126.74,122.89,122.53,122.42,122.07,119.79,119.04,110.45,52.43,52.37.MS(ESI)m/z:451(M+Na)。
Preparation Example 30:
Compound 27:1-(3-(3,4-dimethoxy benzamido) benzyl)-1H-indazole-3-methane amide
Except using 3,4-dimethoxy-benzoyl chloride to replace, outside Benzoyl chloride, preparing compound 27 in the mode identical with Preparation Example 4, be white solid, yield is 82%. 1H NMR(300MHz,DMSO)δ:10.06(s,1H),8.19(d,J=8.2Hz,1H),7.70(m,4H),7.56(d,J=8.4Hz,1H),7.43(m,3H),7.27(m,2H),7.04(d,J=8.4Hz,1H),6.98(d,J=7.8Hz,1H),5.73(s,2H),3.80(s,6H)。 13C NMR(100MHz,dmso)δ:165.00,163.87,151.64,148.26,140.70,139.61,137.64,137.28,128.88,126.83,126.74,122.55,122.48,122.43,122.07,121.06,119.92,119.21,111.03,110.83,110.48,55.66,55.61,52.48.MS(ESI)m/z:457(M+Na)。
Preparation Example 31:
Compound 36:1H-indazole-3-formylaniline
Figure BDA00002060500300201
Except using aniline to replace, ammoniacal liquor, preparing compound 36 in the mode identical with Preparation Example 1, be white solid, yield is 70%. 1H NMR(300MHz,CDCl3)δ：10.47(s,1H),8.91(s,1H),8.47(d,J=8.1Hz,1H),7.75(d,J=8.4Hz,2H),7.43(m,5H),7.15(t,J=7.4Hz,1H).LC-MS m/z(%):260.0(100)[M+Na] +。
Preparation Example 32:
The preparation of compound 37:1-(3-nitrobenzyl)-1H-indazole-3-formylaniline
Except using 1H-indazole-3-formylaniline (compound 36) to replace, 1H-indazole-3-methane amide (compound 33), preparing compound 37 in the mode identical with Preparation Example 2, be white solid, yield is 87%. 1H NMR(300MHz,CDCl 3)δ8.82(s,1H),8.49(d,J=8.1Hz,1H),8.18(s,1H),8.16(s,1H),7.75(d,J=8.1Hz,2H),7.43(m,7H),7.14(t,J=7.5Hz,1H),5.73(s,2H).LC-MS m/z(%):395.0(100)[M+Na] +.
Preparation Example 33:
The preparation of compound 38:1-(3-aminobenzyl)-1H-indazole-3-formylaniline
Figure BDA00002060500300203
By compound 34 (0.372g, 1mmol) be dissolved in 5ml tetrahydrofuran (THF), under room temperature, add two hydrated stannous chloride 1.13g, 67.6mmol) room temperature reaction 12 hours, add 10% aqueous sodium hydroxide solution to adjust pH to alkalescence, ethyl acetate extraction, saturated sodium hydroxide washing, dry, concentrated, column chromatographic isolation and purification obtains compound 38, is white solid (0.256g, productive rate 75%). 1H NMR(300MHz,DMSO)δ10.31(s,1H),8.24(d,J=7.5Hz,1H),7.89(d,J=8.1Hz,2H),7.71(d,J=8.4Hz,1H),7.45(t,J=7.6Hz,1H),7.32(m,3H),7.08(t,J=7.2Hz,1H),6.95(t,J=7.8Hz,1H),6.43(d,J=7.8Hz,2H),6.35(s,1H),5.65(s,2H),5.07(s,2H).LC-MS m/z(%):364.8(100)[M+Na] +。
Preparation Example 34:
Compound 28:1-(3-(3-benzamido) benzyl)-1H-indazole-3-formylaniline
Except using 1-(3-aminobenzyl)-1H-indazole-3-formylaniline (compound 38) to replace 1-(3-aminobenzyl)-1H-indazole-3-methane amide (compound 35), in the mode identical with Preparation Example 4, prepare compound 28, for white solid, yield is 88%. 1H NMR(300MHz,DMSO)δ:10.32(s,1H),10.24(s,1H),8.25(d,J=8.1Hz,1H),7.88(m,4H),7.77(d,J=8.4Hz,1H),7.70(m,2H),7.50(m,4H),7.34(m,4H),7.06(m,2H),5.83(s,2H). 13C NMR(101MHz,DMSO)δ166.23,161.03,141.30,139.95,139.12,138.04,137.70,135.26,132.09,129.46,129.09×2,128.84×2,128.10×2,127.59,124.11,123.35,123.24,123.14,122.37,120.87×2,120.34,119.61,111.17,53.10.MS(ESI)m/z:469(M+Na)。
Pharmacological evaluation embodiment:
1. the inhibition active function of sample to HCV Infection in Vitro human liver tumor cell Huh7.5.1
28 test-compounds are provided by chemosynthesis of the present invention below.
Test method:
Huh7.5.1 cell is inoculated in 96 orifice plates, 37 ℃, 5%CO 2cultivate 24h.With J399EM virus supernatant (moi ≈ 0.1), infect Huh7.5.1 cell, establish FFI cell control well simultaneously, infect after 8 hours, with PBS, clean.The sample that adds different concns in the Huh7.5.1 of J399EM virus infection cell, each concentration is established two multiple holes, and establishes no sample control wells.Given the test agent is since 10 μ M, four times of dilutions, and six kinds of concentration, 10 μ M, 2.5 μ M, 0.625 μ M, 0.0156 μ M, 0.0039 μ M and 0.00098 μ M, add respectively, continues to cultivate 72 hours.After sample preparation 72 hours, on fluorescence microplate reader, excitation wavelength is 488nm, and emission wavelength is 516nm, reads relative intensity of fluorescence (RFU), by formula, calculates HCV virus inhibiting rate.Mix MTT, add MTT lysate after 4 hours, after 6 hours, in microplate reader, 570nm place records OD value.
According to each control of the concentration rate, adopt Logit method calculation of half inhibitory concentration IC 50and half cytotoxicity concentration C C 50.
Table 1: the external HCV virus of compound-J399EM virus suppresses activity and cytotoxicity
Compound CC 50(μM) IC 50(μM)
Compound-1 >10 0.125
Compound-2 >10 0.115
Compound-3 >10 0.238
Compound-4 >10 0.013
Compound-5 >10 0.603
Compound-6 >10 0.346
Compound-7 >10 0.035
Compound-8 >10 0.739
Compound-9 >10 0.213
Compound-10 >10 0.007
Compound-11 >10 0.035
Compound-12 >10 0.230
Compound-13 >10 0.115
Compound-14 >10 0.038
Compound-15 >10 0.012
Compound-16 >10 0.031
Compound-17 6.35 0.044
Compound-18 7.18 0.05
Compound-19 >10 0.053
Compound-20 >10 0.07
Compound-21 >10 0.2
Compound-22 >10 0.53
Compound-23 >10 0.05
Compound-24 >10 1.28
Compound-25 >10 0.11
Compound-26 >10 0.04
Compound-27 >10 1.44
Compound-28 >10 0.382
Note: IC 50concentration while J399EM virus being suppressed to reach half 50% for testing compound;
CC 50for testing compound causes half cytotoxicity desired concn to Huh7.5.1 cell.
2. the inhibition active function of sample to DV2 Infection in Vitro human liver tumor cell Huh7
At DV2, infect after Huh7 cell equally, with PCR in real time method detection compound 4, compound 7, compound 10,11 4 submitted samples of compound are to DV viral RNA level affects effect in culture supernatant, active in 2.
Test method:
Huh7 cell is inoculated in 96 orifice plates, after 24 hours, add DV2 virus infection two hours (MOI=0.05), wash away subsequently virus liquid, change to the compound that adds respectively different concns after fresh culture, establish simultaneously and do not add the infection contrast of compound and normal cell without infecting contrast.Continue to cultivate after 4 days, draw cell culture fluid supernatant through 1000g, 5 minutes centrifugal removing after cell precipitation are used test kit to extract viral RNA from supernatant, with qRT-PCR method, measure the viral genome copy number in supernatant.
Table 2: the external DV virus of compound suppresses activity and cytotoxicity
Note: IC 50concentration while DV virus being suppressed to reach half 50% for testing compound;
CC 50for testing compound causes half cytotoxicity desired concn to Huh7 cell;
NT is undetermined.
3. the inhibition active function of sample to HBV Infection in Vitro
Test method:
The collection of medicine to the inhibition of DNA replication effect culture supernatant of HBV-DNA: HepG2.2.15 cell is cultivated with the MEM nutrient solution containing 10%FBS, 5 * 103 cells/well are inoculated in 96 orifice plates, the medicine that adds different concns next day, establish the control wells that does not add medicine simultaneously, within the 4th day, change nutrient solution and medicine, to be measured in the 8th day collection culture supernatant.Real-time fluorescence quantitative PCR method detects the content of HBV-DNA in culture supernatant: get HepG2.2.15 cells and supernatant, extract HBV-DNA, for pcr amplification, 4 of HBV-DNA standard models are set simultaneously, do typical curve.
Test with PCR primer sequence is:
P1:5’-ATCCTGCTGCTATGCCTCATCTT-3’
P2:5’-ACAGTGGGGAAAGCCCTACGAA-3’
Test with PCR probe sequence is:
5’-TGGCTAGTTTACTAGTGCCATTTTG-3’
According to corresponding HBV-DNA copy number in the culture supernatant of detection gained, calculate the inhibiting rate that example pharmaceuticals copies HB V-DNA, and then carry out example pharmaceuticals half inhibiting rate and calculate, obtain its IC50.
HBV-DNA inhibiting rate %=[1-DNA copy drug treating cell/DNA copy medicine untreated cell] * 100%
Activity the results are shown in Table 3, table 4.
Table 3: the external HBV virus of compound suppresses activity and cytotoxicity
Compound (concentration 100nM) Inhibiting rate % Cell survival rate %
Compound-2 79.3 93.2
Compound-3 66.8 108.7
Compound-4 96.7 18.5
Compound-5 40.9 110.7
Compound-6 42.3 103.7
Compound-7 79.9 82.3
Compound-9 89.2 13.8
Compound-10 40.7 99.6
Compound-11 93.8 71.7
Compound-16 71.8 112.3
Compound-18 36.0 110.1
Compound-19 59.6 60.5
Note: the datumization compound concentration of surveying is 100nM; Inhibiting rate is that external HBV virus suppresses active, the cytotoxicity that cell survival rate is compound; NT is undetermined.
Table 4: compound 4, compound 7, compound 10, the external HBV virus of compound 11 suppress activity and cytotoxicity
Note: IC 50concentration while HBV virus being suppressed to reach half 50% for testing compound;
CC 50for testing compound causes half cytotoxicity desired concn to 2.2.15 cell.
4. the pre-just drug metabolism study of compound 4 and compound 10
Compound 4 dynamics researchs that carry out on rat show, after oral (p.o.) administration, the absorption of compound 4 is very fast, at 1.61h, reaches maximum plasma concentration.Compound 4 Plasma Concentration (C while reaching peak max) be 57ng/mL.Area under the drug-time curve (the AUC of compound 4 0-24h) be respectively 571ngh/mL.The bioavailability of compound 4 (F) is respectively 7.9%.After intravenous injection (i.v.) administration, the transformation period (t of compound 4 1/2) being respectively 0.8h, clearance rate (CL) is respectively 1.31L/ (hkg), Vdss (V ss) be respectively as 1.31L/kg.
Compound 10 dynamics researchs that carry out on rat show, after p.o. administration, compound 10 reaches maximum plasma concentration at 1.7h.Compound 10 Plasma Concentration (C while reaching peak max) be 75.7ng/mL.Area under the drug-time curve (the AUC of compound 10 0-24h) be respectively 748ngh/mL.The bioavailability of compound 10 (F) is respectively 5.25%.After i.v administration, the transformation period (t of compound 10 1/2) being respectively 2.5h, clearance rate (CL) is respectively 0.74L/ (hkg), Vdss (V ss) be respectively as 1.38L/kg.
According to the above results, 1-of the present invention (3-benzamido benzyl)-1H-indazole-3-benzamide type organic micromolecule compound or its pharmacy acceptable salt or hydrate can obviously suppress HBV-DNA in HepG2.2.15 cells and supernatant to be copied, suppress HCV (J399EM virus) and infect Huh7.5.1 cell, suppress DV2 and infect Huh7 cell, and compou nd synthesis is simple, raw material is easy to get, and toxicity is low, can be for the preparation of the medicine of the virus infectiones such as treatment HCV, HBV and DV.
Figure IDA00002060500600011

Claims (10)

1. the 1-shown in general formula 1 (3-benzamido benzyl)-1H-indazole-3-Carbox amide or its pharmacy acceptable salt or a hydrate,
Figure FDA00002060500200011
Wherein, R 1and R 2identical or different, and be H, C independently of one another 1-C 10the alkyl of straight or branched, C 1-C 10the alkoxyl group of straight or branched, C 2-C 10the alkenyl of straight or branched, C 3-C 10cycloalkyl or C 5-C 20aryl;
R 3, R 4, R 5, R 6and R 7identical or different, and be H, halogen, C independently of one another 1-C 10the alkyl of straight or branched, C 1-C 10the alkoxyl group of straight or branched, C 2-C 10the alkenyl of straight or branched, C 3-C 10cycloalkyl, CF 3,-NO 2,-CN, hydroxyl, C 1-C 10alkyl-carbonyl or C 1-C 10alkoxy carbonyl.
2. 1-according to claim 1 (3-benzamido benzyl)-1H-indazole-3-Carbox amide, wherein, R 1and R 2identical or different, and be H, C independently of one another 1-C 6the alkyl of straight or branched, C 1-C 6the alkoxyl group of straight or branched, C 2-C 6the alkenyl of straight or branched, C 3-C 8cycloalkyl or C 5-C 12aryl;
R 3, R 4, R 5, R 6and R 7identical or different, and be H, F, Br, Cl, I, C independently of one another 1-C 6the alkyl of straight or branched, C 1-C 6the alkoxyl group of straight or branched, C 2-C 6the alkenyl of straight or branched, C 3-C 8cycloalkyl, CF 3,-NO 2,-CN, hydroxyl, C 1-C 6alkyl-carbonyl or C 1-C 6alkoxy carbonyl.
3. 1-according to claim 1 (3-benzamido benzyl)-1H-indazole-3-Carbox amide, wherein,
R 1and R 2identical or different, and be H, methyl, ethyl, propyl group, sec.-propyl, butyl, the tertiary butyl, methoxyl group, oxyethyl group, propoxy-, isopropoxy, butoxy, tert.-butoxy, vinyl, propenyl, butenyl, pentenyl, hexenyl, cyclopropane base, tetramethylene base, pentamethylene base, cyclohexyl, suberane base or phenyl independently of one another;
R 3, R 4, R 5, R 6and R 7identical or different, and be H, F, Br, Cl, I, methyl, ethyl, propyl group, sec.-propyl, butyl, the tertiary butyl, methoxyl group, oxyethyl group, propoxy-, isopropoxy, butoxy, tert.-butoxy, vinyl, propenyl, butenyl, pentenyl, hexenyl, cyclopropane base, tetramethylene base, pentamethylene base, cyclohexyl, suberane base, CF independently of one another 3,-NO 2,-CN, hydroxyl, methyl carbonyl, ethyl carbonyl, propyl group carbonyl, sec.-propyl carbonyl, butyl carbonyl, tertiary butyl carbonyl, methoxycarbonyl, ethoxy carbonyl, propoxycarbonyl, isopropoxy carbonyl, butoxy carbonyl or tert-butoxycarbonyl.
4. compound according to claim 1, wherein, described C 5-C 20aromatic base is phenyl, thienyl, furyl, pyridyl, pyrimidyl, pyrryl, imidazolyl, oxazolyl, indazolyl, indyl, quinolyl, naphthyl, isoquinolyl, benzofuryl, benzothienyl or benzothiazolyl.
5. 1-according to claim 1 (3-benzamido benzyl)-1H-indazole-3-Carbox amide, it is selected from following compound:
Figure FDA00002060500200021
Figure FDA00002060500200041
6. a pharmaceutical composition, it comprises and is selected from according to one or more in the 1-described in any one in claim 1 ~ 5 (3-benzamido benzyl)-1H-indazole-3-Carbox amide, its pharmacy acceptable salt and hydrate.
7. prepare the method for 1-according to claim 1 (3-benzamido benzyl)-1H-indazole-3-Carbox amide, it comprises the steps:
Reaction stream formula 1
A) take 3-formic acid-1H-indazole is initiator, and under the effect of thionyl chloride or oxalyl chloride, reaction generates after acyl chlorides and amine HNR again 1r 2there is amidate action and obtain the compound shown in general formula 2;
B) there is the compound shown in N-1 position substitution reaction generation general formula 3 with 1-(bromomethyl)-3-oil of mirbane in the compound shown in general formula 2 under the effect of alkali;
C) then the compound shown in general formula 3 is reduced to the compound shown in general formula 4 under the effect of reductive agent;
D) compound shown in general formula 4 under amide condensed dose of effect with or with
Figure FDA00002060500200052
there is amidate action and generate the compound shown in general formula 1,
Wherein, R 1, R 2, R 3, R 4, R 5, R 6and R 7definition identical with the definition in claim 1.
8. method according to claim 7, wherein,
Step b) alkali in is selected from sodium carbonate, salt of wormwood, sodium hydroxide, potassium hydroxide, sodium methylate, sodium ethylate, sodium ethylate, potassium ethylate, sodium hydride and potassium hydride KH;
Step c) reductive agent in is selected from tindichloride, iron powder and zinc powder;
Steps d) amide condensed dose in is selected from carbonyl dimidazoles, dicyclohexylcarbodiimide, DIC, 1-(3-dimethylamino-propyl)-3-ethyl carbodiimide, 2-(7-azo benzotriazole)-N, N, N', N'-tetramethyl-urea phosphofluoric acid ester, benzotriazole-N, N, N', N'-tetramethyl-urea hexafluorophosphate, 6-Chloro-Benzotriazole-1, 1, 3, 3-tetramethyl-urea hexafluorophosphate, O-benzotriazole-N, N, N', N'-tetramethyl-urea Tetrafluoroboric acid ester, 6-Chloro-Benzotriazole-1, 1, 3, 3-tetramethyl-urea Tetrafluoroboric acid ester, 2-succinimido-1, 1, 3, 3-tetramethyl-urea Tetrafluoroboric acid ester and 2-(5-norbornylene-2, 3-dicarboximide base)-1, 1, 3, 3-tetramethyl-urea tetrafluoroborate.
9. according to the compound described in any one in claim 1 ~ 5, or pharmacy acceptable salt or the purposes of hydrate in the medicine for the preparation for the treatment of virus infection.
10. purposes according to claim 9, wherein, described virus is hepatitis C virus.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103922966A (en) * 2014-04-14 2014-07-16 武汉大学 Formamide and isonitrile compounds serving as influenza A virus inhibitors and preparation and application thereof
WO2018133845A1 (en) * 2017-01-23 2018-07-26 上海长森药业有限公司 Thiourea and urea compound and use thereof
EP4076444A4 (en) * 2019-12-18 2024-01-10 Merck Sharp & Dohme Llc Indazole derivatives and methods of use thereof for the treatment of herpes viruses

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003174898A (en) * 2001-08-20 2003-06-24 Japan Science & Technology Corp Method for retrieving intracellular antiviral factor
WO2007147336A1 (en) * 2006-06-13 2007-12-27 Shanghai Institue Of Materia Medica, Chinese Academy Of Sciences Heterocyclic non-nucleoside compounds, their peparation, pharmaceutical composition and their use as antiviral agents
CN102151256A (en) * 2011-02-23 2011-08-17 中国农业大学 Application of protocatechuic acid in preparation of drugs for preventing and controlling livestock and poultry virus infectious diseases
CN102219725A (en) * 2010-04-16 2011-10-19 中国科学院上海药物研究所 Benzohetercyclic compound as well as preparation method and applications thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003174898A (en) * 2001-08-20 2003-06-24 Japan Science & Technology Corp Method for retrieving intracellular antiviral factor
WO2007147336A1 (en) * 2006-06-13 2007-12-27 Shanghai Institue Of Materia Medica, Chinese Academy Of Sciences Heterocyclic non-nucleoside compounds, their peparation, pharmaceutical composition and their use as antiviral agents
CN102219725A (en) * 2010-04-16 2011-10-19 中国科学院上海药物研究所 Benzohetercyclic compound as well as preparation method and applications thereof
CN102151256A (en) * 2011-02-23 2011-08-17 中国农业大学 Application of protocatechuic acid in preparation of drugs for preventing and controlling livestock and poultry virus infectious diseases

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103922966A (en) * 2014-04-14 2014-07-16 武汉大学 Formamide and isonitrile compounds serving as influenza A virus inhibitors and preparation and application thereof
CN103922966B (en) * 2014-04-14 2015-10-21 武汉大学 As the methane amide of influenza A virus inhibitor and isonitrile compounds and preparation and application thereof
WO2018133845A1 (en) * 2017-01-23 2018-07-26 上海长森药业有限公司 Thiourea and urea compound and use thereof
CN108341817A (en) * 2017-01-23 2018-07-31 上海长森药业有限公司 Thiourea, carbamide compounds and application thereof
CN108341817B (en) * 2017-01-23 2021-11-26 上海长森药业有限公司 Thiourea, urea compound and use thereof
EP4076444A4 (en) * 2019-12-18 2024-01-10 Merck Sharp & Dohme Llc Indazole derivatives and methods of use thereof for the treatment of herpes viruses

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