CN101830854A - Substituted phenylhydrazones compounds, preparation method and application thereof - Google Patents

Substituted phenylhydrazones compounds, preparation method and application thereof Download PDF

Info

Publication number
CN101830854A
CN101830854A CN200910047268A CN200910047268A CN101830854A CN 101830854 A CN101830854 A CN 101830854A CN 200910047268 A CN200910047268 A CN 200910047268A CN 200910047268 A CN200910047268 A CN 200910047268A CN 101830854 A CN101830854 A CN 101830854A
Authority
CN
China
Prior art keywords
compound
polysubstituted
preparation
formula
diazanyl
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.)
Granted
Application number
CN200910047268A
Other languages
Chinese (zh)
Other versions
CN101830854B (en
Inventor
朱维良
左之利
李剑
刘爱华
邓菁
朱进
张碧贞
李秀慧
陈刚
潘春木
陈凯先
蒋华良
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
East China University of Science and Technology
Shanghai Institute of Materia Medica of CAS
Singapore Polytechnic
Original Assignee
East China University of Science and Technology
Shanghai Institute of Materia Medica of CAS
Singapore Polytechnic
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by East China University of Science and Technology, Shanghai Institute of Materia Medica of CAS, Singapore Polytechnic filed Critical East China University of Science and Technology
Priority to CN200910047268XA priority Critical patent/CN101830854B/en
Publication of CN101830854A publication Critical patent/CN101830854A/en
Application granted granted Critical
Publication of CN101830854B publication Critical patent/CN101830854B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Landscapes

  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention relates to substituted phenylhydrazones compounds, a preparation method and application thereof, in particular to novel multi-substituted single or dual phenylhydrazones compounds, a preparation method and application thereof in preparing drugs for treating singapore hemorrhagic fever and the like caused by dengue virus. A structural formula of the compounds is shown in the specification. The compounds can be used as a dengue virus NS2B-NS3 protease inhibitor, thereby preventing further proliferation of the dengue virus in a host.

Description

The substituted phenylhydrazones compounds thing, and its production and use
Technical field
The present invention relates to pharmaceutical chemistry and pharmacotherapeutics field, more specifically relate to polysubstituted single or two phenyl hydrazones compound and preparation method thereof.This compounds can be used for treating the disease that dengue virus (DV, Dengue virus) causes.
Background technology
Dengue virus is the important member that flaviviridae (flaviviridae) Flavivirus (flavivirus) is stepped on the leather subgroup.It is a single-stranded RNA virus, and the tool infectivity constitutes viral nucleocapsid with alkaline capsid protein c.The dengue virus that occurring in nature exists can be divided into 4 serum, is respectively DEN-1, DEN-2, DEN-3 and DEN-4, has antigenic cross-reaction between different serotypes.Can cause that slighter singapore hemorrhagic fever (DF, Dengue fever) and extremely serious stepping on expel blood dengue/dengue shock syndrome (DHF/DSS, Dengue hemorrhagic fever/Dengue shock syndrome).Its communication media is Aedes aegypti and Aedes albopictus, and Susceptible population is children.
It is reported that singapore hemorrhagic fever is widely current in the torrid zone and semi-tropical more than 100 countries and regions, there are 0.5～100,000,000 DF and 25～500,000 DHF patients in the whole world every year, and 5000 people death owing to DHF/DSS is wherein arranged approximately.All once there was the generation of DF in provinces such as the Guangdong of China, Hainan, Taiwan or was very popular, it is one of the highest arthropod borne viral disease of China's sickness rate, and DHF and DSS have very high mortality ratio, become a serious public health problem so DV propagates the harm that is caused.
Popular research direction to dengue virus comprises dengue virus infection Mechanism Study (cell levels research and virus levels research), dengue virus infection diagnostic method, the research of stepping on the leather vaccine, the structure of animal model, and anti-research of stepping on the leather medicine at present.At present, only depend on control mosquito matchmaker's propagation to prevent the popular of singapore hemorrhagic fever, it is still at the experimental stage to step on the leather vaccine safely and effectively, and there also do not have the special efficacy antiviral to be applied to be clinical, mainly takes symptomatic treatment and supportive treatment.Though anti-DV is effective in existing antiviral in vitro tests, the in vivo test effect is neither obviously.In this case, demand researching and developing anti-dengue virus medicine urgently with remarkable therapeutic action.
Duplicating of dengue fever virus is polyprotein of rna gene group coding by its 11kb, under the effect of virus and host's proteolytic enzyme, this polyprotein enzyme is resolved into the albumen with difference in functionality, comprise three structural protein (C, prM and E) and seven Nonstructural Protein (NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5).NS3 albumen is second largest viral protein in these ten kinds of albumen, has four kinds of different enzymic activitys: serine protease, rna helicase enzyme, ntp protein enzyme and RNA triphosphatase.The protease activity of NS3 and helicase activity are very important to virus replication, and this makes NS3 albumen become a potential anti-dengue virus target.Studies show that further NS2B albumen plays crucial effects as the cofactor of NS3 proteolytic enzyme to NS3 proteolytic enzyme performance enzymic activity, only after both form the dimer mixture, could activate NS3 proteolytic enzyme.
In recent years, " area of computer aided medicinal design " (Computer-Aided Drug Design CADD) has become the important method and the instrument of modern medicines research and development.The medicinal design of utilization area of computer aided can be shortened 0.9 year new drug research cycle on average, saves 1.3 hundred million dollars of research and development costs.Therefore, the CADD method has been widely used in the discovery and the optimization of lead compound.The area of computer aided medicinal design has become a kind of area of computer aided drug discovery method commonly used at present.Its principle be the utilization supercomputer by the method for molecular docking in reaching millions of micromolecular compound storehouses, seeks can with a certain specified disease target proteins bonded chemical structure, for further molecule, cell, animal and clinical study provide high-efficiency reliable small molecules lead compound.
In sum, be target with dengue virus NS2B-NS3 proteolytic enzyme, in conjunction with the biological activity test test of CADD and various levels, the micromolecular compound that designs and synthesizes has important practical significance to developing the anti-leather medicine of stepping on.
Integrated use CADD of the present invention and enzyme inhibition activity experiment have found that the two phenyl hydrazones compounds of replacement have the dengue virus of inhibition NS2B-NS3 protease activities.The present invention further uses the rational drug method of design to carry out structural modification at this compound and has obtained the brand-new substituted benzene hydrazone compounds of a class formation, and test this compounds and in the horizontal experiment test of vitro enzyme, suppress dengue virus NS2B-NS3 protease activities, the result proves that the application's compound has stronger dengue virus NS2B-NS3 protease inhibiting activity.
Summary of the invention
The present invention has designed the polysubstituted single or two phenyl hydrazones compound of a class with dengue virus NS2B-NS3 proteolytic enzyme for target, has invented its preparation method, and has measured the ability of this compounds inhibition dengue virus NS2B-NS3 protease activity.
An object of the present invention is to provide the novel polysubstituted single or two phenyl hydrazones compound with the effect of anti-dengue virus NS2B-NS3 proteolytic enzyme, its general structure as the formula (1).
Another object of the present invention provides the preparation method of polysubstituted single or two phenyl hydrazones compound.
A further object of the present invention provides as the application in the pharmaceutical composition of polysubstituted single or two phenyl hydrazones compound aspect the relative disease that preparation prevents or the treatment dengue virus causes of dengue virus NS2B-NS3 proteinase inhibitor.
The present invention also provides and comprises one or more the above-mentioned polysubstituted single or two phenyl hydrazones compounds for the treatment of significant quantity and the pharmaceutical composition of pharmaceutically acceptable carrier.
Polysubstituted single or two phenyl hydrazones compound involved in the present invention can be used as the micromolecular inhibitor of dengue virus NS2B-NS3 proteolytic enzyme, suppresses duplicating of dengue virus by suppressing the NS2B-NS3 protease activity, thus the life cycle of blocking-up dengue virus.Therefore can be developed into and be new anti-dengue virus medicine, can be used for preventing or treating all kinds of diseases that dengue virus causes, as singapore hemorrhagic fever (dengue fever, DF), dengue hemorrhagic fever (dengue hemorrhagic fever, DHF) and dengue shock syndrome (dengue shock syndrome, DSS).
The invention provides polysubstituted single or two phenyl hydrazones compound with following general formula (1) structure:
Figure B200910047268XD0000041
Wherein:
R 1Be selected from H ,-NO 2With the C1-C4 alkoxy carbonyl;
R 2Be selected from H and-OH;
R 3Be selected from the saturated or unsaturated C1-C10 alkyl, ArC (O) of straight or branched-, Ar-,
Figure B200910047268XD0000042
Figure B200910047268XD0000043
With
Figure B200910047268XD0000044
Perhaps R 3Form 1,3-dihydro-1,3-dioxo-2H-pseudoindoyl with the N that is connected;
R 4Be selected from the C1-C4 alkyl ,-COOH and
Figure B200910047268XD0000045
R 5Be selected from the saturated or unsaturated C1-C4 alkyl and the Ar-of H, straight or branched;
Ar is selected from replacement or unsubstituted phenyl and contains 1～3 and is selected from N; the heteroatomic 5-7 membered aromatic heterocycle base of O and S; and described 5-7 membered aromatic heterocycle base is not necessarily by phenyl and close, and substituting group can be 1～4 and be independently selected from halogen; C1-C6 straight or branched alkyl; hydroxyl; the C1-C4 alkoxyl group; the unsaturated-oxyl of C1-C4; carboxyl; ester group; C1-C6 carboxyl alkoxyl group; C1-C6 ester group alkoxyl group; the C1-C6 carboxyalkyl; C1-C6 ester group alkyl; cyano group; nitro; amino; methylol; trifluoromethyl; trifluoromethoxy; sulfydryl; sulfamyl; the group of dioxolane and C1-C4 acyl group.
Preferably, in the above-mentioned general formula (1):
R 1Be selected from H ,-NO 2With-COOC 2H 5
R 2Be selected from H and-OH;
R 3Be selected from the saturated or unsaturated C1-C4 alkyl, ArC (O) of straight or branched-, Ar-,
Figure B200910047268XD0000051
With
Figure B200910047268XD0000053
Perhaps R 3Form 1,3-dihydro-1,3-dioxo-2H-isoindole-2-base with the N that is connected;
R 4Be selected from-CH 3,-COOH and
Figure B200910047268XD0000054
R 5Be selected from H ,-CH 3,-C 2H 5,-CH 2-CH=CH 2And Ar-;
Ar is selected from and replaces or unsubstituted phenyl, benzimidazolyl-and pyridyl, and substituting group can be 1～4 group that is independently selected from C1-C6 straight or branched alkyl, C1-C4 alkoxyl group, nitro, dioxolane and sulfamyl.
Further, first preferred embodiment of formula of the present invention (1) compound is a Compound I A,
Wherein:
R 1Be selected from-COOC2 XH 5
R 2Be selected from-OH;
R 3, R 4And R 5Definition same as described above;
Second preferred embodiment of formula of the present invention (1) compound is Compound I B,
Wherein:
R 1Be selected from-NO 2
R 2Be selected from-OH;
R 4Be selected from-CH 3
R 3And R 5Definition same as described above;
The 3rd preferred embodiment of formula of the present invention (1) compound is Compound I C,
Wherein:
R 1Be selected from H;
R 2Be selected from-OH;
R 4Be selected from-CH 3With
R 3And R 5Definition same as described above.
Further preferably, above-mentioned general formula (1) compound is specially:
Figure B200910047268XD0000062
Figure B200910047268XD0000071
Figure B200910047268XD0000081
Figure B200910047268XD0000091
The present invention also provides polysubstituted single or two phenyl hydrazones compound (I of general formula (1) structure A～I C) and the preparation method of intermediate II～XII, concrete synthesis strategy is as follows respectively.
I ASynthetic:
Figure B200910047268XD0000092
1) with 4-hydroxyl-3, the 5-mesitylenic acid is put in the dehydrated alcohol, adds an amount of acid catalyst (as the vitriol oil, concentrated hydrochloric acid and Glacial acetic acid), stirred 10-20 hour in 60-80 ℃, steaming desolventizes, and obtains 4-hydroxyl-3,5-mesitylenic acid ethyl ester (intermediate II).
2) intermediate II and acetic anhydride are mixed in 120-140 ℃ stirred 10-20 hour, steaming desolventizes, and obtains the 4-acetoxy-3,5-mesitylenic acid ethyl ester (intermediate III).
3) intermediate III is joined in an amount of isopyknic Glacial acetic acid and acetic anhydride mixed solution, under temperature control 0-5 ℃, drip an amount of vitriol oil, add excessive chromium trioxide (5-10 equivalent) afterwards again, about 2-4 of time spent hour in batches.Keep temperature of reaction to continue to stir 24-48 hour, in reaction solution impouring frozen water, add Sodium Metabisulfite cancellation reaction, stirring at room 15-30 minute.With organic solvent (as ether) extraction, drying, steaming desolventizes.Add an amount of ethanol and the catalytic amount vitriol oil to resistates, backflow 3-5 hour.Steaming desolventizes; residual solid is separated through silica gel column chromatography; by the ascending order of polarity; obtain 3-formyl radical-4-hydroxy-5-methyl yl benzoic acid ethyl ester (intermediate compound IV), 3 successively, 5-diformyl-4-hydroxy-benzoic acid ethyl ester (intermediate V) and 2-hydroxyl-3-formyl radical-5-ethoxycarbonyl-phenylformic acid (intermediate VI).
4) with three intermediate compound IV, V, VI respectively with R 3The hydrazine that replaces is put in the dehydrated alcohol, adds the Glacial acetic acid of catalytic amount, in 60-80 ℃ of stirring 2-5 hour, and suction filtration, ethanol is washed, and obtains three Compound I respectively A, promptly 3-replaces diazanyl methene base-4-hydroxy-5-methyl yl benzoic acid ethyl ester, 3, and two (replacing diazanyl methene the base)-4-hydroxy-benzoic acid ethyl esters of 5-and 2-hydroxyl-3-replace diazanyl methene base-5-ethoxycarbonyl-phenylformic acid.
Wherein, R 3And R 4Definition the same.
I BSynthetic:
Figure B200910047268XD0000111
1) with 2, the 6-xylenol is dissolved in an amount of Glacial acetic acid, temperature control 0-5 ℃, splash into the mixing solutions (V/V=1/3) of concentrated nitric acid and Glacial acetic acid, keep temperature of reaction to continue to stir 2-5 hour, in reaction solution impouring frozen water, suction filtration, washing solid, drying, obtain 4-nitro-2,6-xylenol (intermediate VII).
2) with intermediate VII with above-mentioned I ASynthetic in the 2nd) similar method of step prepares intermediate VIII O-(4-nitro-2,6-3,5-dimethylphenyl) acetic ester;
3) with above-mentioned I ASynthetic in the 3rd), 4) go on foot similar method, prepare Compound I B, promptly 2-replaces diazanyl methene base-4-nitro-6-methyl-phenol.
Wherein, R 3Definition the same.
I CSynthetic:
Figure B200910047268XD0000121
1) with 2, the 6-xylenol is with above-mentioned I ASynthetic in the 2nd) similar method of step prepares O-(2, the 6-3,5-dimethylphenyl) acetic ester (intermediate X).
2) with above-mentioned I ASynthetic in the 3rd), 4) go on foot similar method, prepare Compound I C, promptly 2-replaces diazanyl methene base-6-methyl-phenol and 2, two (replacing diazanyl methene the base)-phenol of 6-.
R wherein 4Be selected from-CH 3With
Figure B200910047268XD0000122
R 3Definition as mentioned above.
Description of drawings
Fig. 1 is a Compound I A-2 couples of amount effect curve figure that the NS2B-NS3 enzyme is lived and suppressed.
Fig. 2 is a Compound I A-5 couples of amount effect curve figure that the NS2B-NS3 enzyme is lived and suppressed.
Fig. 3 is a Compound I A-7 couples of amount effect curve figure that the NS2B-NS3 enzyme is lived and suppressed.
Fig. 4 is a Compound I A-9 couples of amount effect curve figure that the NS2B-NS3 enzyme is lived and suppressed.
Fig. 5 is a Compound I A-14 couples of amount effect curve figure that the NS2B-NS3 enzyme is lived and suppressed.
Fig. 6 is a Compound I A-16 couples of amount effect curve figure that the NS2B-NS3 enzyme is lived and suppressed.
Fig. 7 is a Compound I C-2 couples of amount effect curve figure that the NS2B-NS3 enzyme is lived and suppressed.
Embodiment
In following embodiment, will further illustrate the present invention.These embodiment only are used to illustrate the present invention, but do not limit the present invention in any way.All parameters among the embodiment and remaining explanation unless otherwise indicated, all are to be unit with quality (gram).
Embodiment 14-hydroxyl-3, the preparation of 5-mesitylenic acid ethyl ester (intermediate II)
Figure B200910047268XD0000131
With 0.50 gram 4-hydroxyl-3, the 5-mesitylenic acid is put in 20 milliliters of dehydrated alcohols, adds 1 milliliter of vitriol oil, refluxes and stirs 15 hours, and steaming desolventizes, and obtains 0.54 gram 4-hydroxyl-3,5-mesitylenic acid ethyl ester (intermediate II).Be white solid, yield 92%.Mp?115-116℃； 1H?NMR(CDCl 3，500MHz)δ1.36(t，3H)，2.26(s，6H)，4.34(q，2H)，7.72(s，2H)。
Embodiment 24-acetoxy-3, the preparation of 5-mesitylenic acid ethyl ester (intermediate III)
Figure B200910047268XD0000132
With 0.16 gram 4-hydroxyl-3,5-mesitylenic acid ethyl ester (intermediate II) and 5 milliliters of acetic anhydride mix stirring 18 hours in 140 ℃, and steaming desolventizes, and obtains 0.16 and restrains 4-acetoxy-3,5-mesitylenic acid ethyl ester (intermediate III).Be white solid, yield 82%. 1H?NMR(CDCl 3，500MHz)δ1.36(t，3H)，2.18(s，6H)，2.34(s，3H)，4.34(q，2H)，7.75(s，2H)。
Embodiment 33-formyl radical-4-hydroxy-5-methyl yl benzoic acid ethyl ester (intermediate compound IV), 3, the preparation of 5-diformyl-4-hydroxy-benzoic acid ethyl ester (intermediate V) and 2-hydroxyl-3-formyl radical-5-ethoxycarbonyl-phenylformic acid (intermediate VI)
Figure B200910047268XD0000141
With 0.35 gram 4-acetoxy-3,5-mesitylenic acid ethyl ester (intermediate III) joins in 15 milliliters the isopyknic Glacial acetic acid and acetic anhydride mixed solution, under 0 ℃ of the temperature control, drips 1 milliliter the vitriol oil, the chromium trioxide (5 equivalent) that in batches adds 0.69 gram afterwards again, about 2 hours of time spent.Keep temperature of reaction to continue to stir 20 hours, in reaction solution impouring frozen water, add 10 gram Sodium Metabisulfite cancellation reactions, stirring at room 15 minutes.Use extracted with diethyl ether, drying, steaming desolventizes.Add 10 milliliters of ethanol to resistates, 3 vitriol oils refluxed 4 hours.Steaming desolventizes, and residual solid is separated through silica gel column chromatography, and eluent is ethyl acetate/petroleum ether=1/6, by the ascending order of polarity, obtains successively:
25 milligrams of 3-formyl radical-4-hydroxy-5-methyl yl benzoic acid ethyl esters (intermediate compound IV) are yellow solid, yield 8%. 1H?NMR(CDCl 3，500MHz)δ1.40(t，3H)，2.25(s，3H)，4.40(q，2H)，8.04(s，1H))，8.15(s，1H)，9.95(s，1H)。
38 milligram 3,5-diformyl-4-hydroxy-benzoic acid ethyl ester (intermediate V) is yellow solid, yield 11%. 1H?NMR(CDCl 3，500MHz)δ1.36(t，3H)，4.40(q，2H)，8.52(s，2H)，10.30(s，2H)；MS(EI)m/z?222.1[M] +。
35 milligrams of 2-hydroxyl-3-formyl radical-5-ethoxycarbonyl-phenylformic acid (intermediate VI) are red solid, yield 10%. 1H?NMR(CDCl 3，500MHz)δ1.40(t，3H)，4.40(q，2H)，8.68(s，1H)，8.87(s，1H)，10.42(s，1H)；MS(EI)m/z?238.1[M] +。
Embodiment 43-(2-(1H-benzimidazolyl-2 radicals-yl) diazanyl methene base)-4-hydroxy-5-methyl yl benzoic acid ethyl ester (Compound I A-1) preparation
Figure B200910047268XD0000151
(the 1H-benzimidazolyl-2 radicals-yl) hydrazine is put into respectively in 5 milliliters of dehydrated alcohols with 69 milligrams of 1-with 75 milligrams of 3-formyl radicals-4-hydroxy-5-methyl yl benzoic acid ethyl ester (intermediate compound IV); add 3 Glacial acetic acid; stirred 2 hours in 80 ℃; suction filtration; ethanol is washed, and obtains 80 milligrams of 3-(2-(1H-benzimidazolyl-2 radicals-yl) diazanyl methene base)-4-hydroxy-5-methyl yl benzoic acid ethyl ester (Compound I A-1), is yellow solid, yield 65%.Mp?279-282℃； 1H?NMR(DMSO，500MHz)δ1.31(t，3H)，2.28(s，3H)，4.27(q，2H)，6.96(d，2H)，7.13(d，2H)，7.71(s，1H)，7.87(s，1H)，8.37(s，1H)；MS(EI)m/z?338.1[M] +。
Embodiment 53, the two (2-(1H-benzimidazolyl-2 radicals-yl) diazanyl methene base)-4-hydroxy-benzoic acid ethyl ester (Compound I of 5- A-2) preparation
Figure B200910047268XD0000152
With 75 milligram 3; (the 1H-benzimidazolyl-2 radicals-yl) put into respectively in 5 milliliters of dehydrated alcohols by hydrazine with 110 milligrams of 1-for 5-diformyl-4-hydroxy-benzoic acid ethyl ester (intermediate V); add 2 Glacial acetic acid; stirred 2 hours in 80 ℃; suction filtration; ethanol is washed, and obtains 36 milligram 3, the two (2-(1H-benzimidazolyl-2 radicals-yl) diazanyl methene base)-4-hydroxy-benzoic acid ethyl ester (I of 5- A-2), yield 22%.Be yellow solid, Mp＞300 ℃; 1HNMR (DMSO, 500MHz) δ 1.37 (t, 3H), 4.36 (q, 2H), 7.00 (s, 4H), 7.22 (s, 4H), 8.25 (s, 2H), 8.42 (s, 2H); MS (ESI) m/z 505.2[M+Na] +
Embodiment 62-hydroxyl-3-(2-(1H-benzimidazolyl-2 radicals-yl) diazanyl methene base)-5-ethoxycarbonyl phenylformic acid (Compound I A-3) preparation
(the 1H-benzimidazolyl-2 radicals-yl) hydrazine is put into respectively in 5 milliliters of dehydrated alcohols with 60 milligrams of 1-with 74 milligrams of 2-hydroxyls-3-formyl radical-5-ethoxycarbonyl-phenylformic acid (intermediate VI); add 2 Glacial acetic acid; stirred 2 hours in 80 ℃; suction filtration; ethanol is washed, and obtains 8 milligrams of 2-hydroxyl-3-(2-(1H-benzimidazolyl-2 radicals-yl) diazanyl methene base)-5-ethoxycarbonyl phenylformic acid (I A-3), yield 7%.Be yellow solid, Mp＞300 ℃; 1H NMR (DMSO, 500MHz) δ 1.37 (t, 3H), 4.32 (q, 2H), 7.80 (s, 1H), 7.85 (m, 2H), 7.87 (m, 2H), 8.53 (s, 1H), 9.72 (s, 1H); MS (EI) m/z 368.3[M] +
Embodiment 73, two (4-oil of mirbane diazanyl methene the base)-4-hydroxy-benzoic acid ethyl ester (Compound I of 5- A-4) preparation
Figure B200910047268XD0000162
Except (the 1H-benzimidazolyl-2 radicals-yl) hydrazine replaces to the 4-nitrophenyl hydrazine, and all the other desired raw materials, reagent and preparation method get Compound I with embodiment 5 with 1- A-4,26 milligrams of yellow solids, yield 24%.Mp＞300℃； 1H?NMR(DMSO，500MHz)δ1.37(t，3H)，4.37(q，2H)，7.14(d，4H)，8.20(d，4H)，8.24(s，2H)，8.43(s，2H)；MS(EI)m/z?492.1[M] +。
Embodiment 83, the two (preparations of 2-(1H-benzimidazolyl-2 radicals-yl) diazanyl methene base)-ethyl benzoate (Compound I A-5) of 5-
Figure B200910047268XD0000171
Except with 4-hydroxyl-3, the 5-mesitylenic acid replaces to 3, and outside the 5-mesitylenic acid, all the other desired raw materials, reagent and preparation method make 3 with embodiment 1,5-mesitylenic acid ethyl ester; According to embodiment 3 described operations, make 3,5-diformyl-ethyl benzoate subsequently; According to embodiment 5 described operations, make Compound I again A-5,42 milligrams of yellow solids, yield 37%.Mp＞300℃； 1H?NMR(DMSO，500MHz)δ1.37(t，3H)，4.32(q，2H)，7.00(m，4H)，7.27(m，4H)，8.15(s，1H)，8.17(s，1H)，8.26(s，1H)；MS(EI)m/z?466.2[M] +。
Embodiment 93, two ((1,3-dihydro-1,3-dioxo-2H-isoindole-2-yl) diazanyl methene the base)-4-hydroxy-benzoic acid ethyl ester (Compound I of 5- A-6) preparation
Except (the 1H-benzimidazolyl-2 radicals-yl) hydrazine replaces to the N-aminophthalimide, and all the other desired raw materials, reagent and preparation method get title compound, 47 milligrams of yellow solids, yield 60% with embodiment 5 with 1-.Mp?250-255℃； 1H?NMR(DMSO，500MHz)δ1.37(t，3H)，4.32(q，2H)，7.82(m，2H)，7.91(m，4H)，7.97(m，4H)，8.57(s，2H)；MS(EI)m/z?510.2[M] +。
Embodiment 103, two (2-nicotinoyl diazanyl methene the base)-4-hydroxy-benzoic acid ethyl ester (Compound I of 5- A-7) preparation
Figure B200910047268XD0000181
Except (the 1H-benzimidazolyl-2 radicals-yl) hydrazine replaces to the nicotinoyl hydrazine, and all the other desired raw materials, reagent and preparation method get title compound, 43 milligrams of yellow solids, yield 49% with embodiment 5 with 1-.Mp?169-170℃； 1H?NMR(DMSO，500MHz)δ1.37(t，3H)，4.32(q，2H)，7.60(q，3H)，8.27(d，2H)，8.35(s，2H)，8.79(d，3H)，9.12(s，2H)；MS(EI)m/z?460.2[M] +。
Embodiment 113, two (the different nicotinoyl diazanyl of 2-methene the base)-4-hydroxy-benzoic acid ethyl ester (Compound I of 5- A-8) preparation
Figure B200910047268XD0000182
Except (the 1H-benzimidazolyl-2 radicals-yl) hydrazine replaces to the different nicotinoyl hydrazine, and all the other desired raw materials, reagent and preparation method get title compound, 19 milligrams of yellow solids, yield 17% with embodiment 5 with 1-.Mp?169-170℃； 1H?NMR(DMSO，500MHz)δ1.37(t，3H)，4.32(q，2H)，7.70(q，1H)，7.82(d，4H)，8.37(s，2H)，8.68(q，1H)，8.81(d，4H)；MS(EI)m/z?460.2[M] +。
Embodiment 123, two (4-sulfamyl diazanyl methene the base)-4-hydroxy-benzoic acid ethyl ester (Compound I of 5- A-9) preparation
Figure B200910047268XD0000191
Except (the 1H-benzimidazolyl-2 radicals-yl) hydrazine replaces to the 4-diazanyl benzsulfamide, and all the other desired raw materials, reagent and preparation method get title compound, 70 milligrams of yellow solids, yield 82% with embodiment 5 with 1-.Mp?242-248℃； 1H?NMR(DMSO，500MHz)δ1.37(t，3H)，4.32(q，2H)，7.11(d，4H)，7.73(d，4H)，8.22(s，2H)，8.35(s，2H)；MS(ESI)m/z?583.1[M+Na] +。
Embodiment 133, two (2-(carbaniloyl,phenylcarbamoyl) diazanyl methene the base)-4-hydroxy-benzoic acid ethyl ester (Compound I of 5- A-10) preparation
Figure B200910047268XD0000192
Except (the 1H-benzimidazolyl-2 radicals-yl) hydrazine replaces to the 4-Carbaphen, and all the other desired raw materials, reagent and preparation method get title compound, 71 milligrams of yellow solids, yield 75% with embodiment 5 with 1-.Mp＞300℃； 1H?NMR(DMSO，500MHz)δ1.37(t，3H)，4.32(q，2H)，7.03(t，2H)，7.31(t，4H)，7.58(d，4H)，8.33(s，2H)，8.38(s，2H)；MS(ESI)m/z?511.2[M+Na] +。
Embodiment 143, the preparation of two (2-amidino groups diazanyl methene the base)-4-hydroxy-benzoic acid ethyl esters (Compound I A-11) of 5-
Figure B200910047268XD0000201
Except with 1-(hydrazine of 1H-benzimidazolyl-2 radicals-yl) replaces to outside the Urea,amino-, and all the other desired raw materials, reagent and preparation method be with embodiment 5, title compound, 39 milligrams of yellow solids, yield 54%.Mp194-198℃； 1H?NMR(DMSO，500MHz)δ1.37(t，3H)，4.32(q，2H)，7.24(s，2H)，7.72(s，2H)，8.31(s，6H)，8.42(s，2H)；MS(ESI)m/z?357.2[M+Na] +。
Embodiment 153, two (2-(methylamino-thiocarbonyl group) diazanyl methene the base)-4-hydroxy-benzoic acid ethyl ester (Compound I of 5- A-12) preparation
Figure B200910047268XD0000202
Except (the 1H-benzimidazolyl-2 radicals-yl) hydrazine replaces to the 4-methylamino thiocarbamide, and all the other desired raw materials, reagent and preparation method get title compound, 29 milligrams of yellow solids, yield 56% with embodiment 5 with 1-.Mp?254-259℃； 1H?NMR(DMSO，500MHz)δ1.37(t，3H)，4.32(q，2H)，3.02(d，6H)，8.27(s，2H)，8.41(s，2H)，8.60(s，2H)；MS(EI)m/z?396.4[M] +。
Embodiment 163, two (2-(ethylamino thiocarbonyl group) diazanyl methene the base)-4-hydroxy-benzoic acid ethyl ester (Compound I of 5- A-13) preparation
Figure B200910047268XD0000211
Except (the 1H-benzimidazolyl-2 radicals-yl) hydrazine replaces to the 4-ethylamino thiocarbamide, and all the other desired raw materials, reagent and preparation method get title compound, 35 milligrams of yellow solids, yield 43% with embodiment 5 with 1-.Mp?263-265℃； 1H?NMR(DMSO，500MHz)δ1.37(t，3H)，3.44(q，6H)，3.60(t，4H)，4.34(q，2H)，8.27(s，2H)，8.42(s，2H)，8.64(s，2H)；MS(EI)m/z424.2[M] +。
Embodiment 173, two (2-(thiocarbamoyl) diazanyl methene the base)-4-hydroxy-benzoic acid ethyl ester (Compound I of 5- A-14) preparation
Figure B200910047268XD0000212
Except (the 1H-benzimidazolyl-2 radicals-yl) hydrazine replaces to the thiosemicarbazide, and all the other desired raw materials, reagent and preparation method get title compound, 44 milligrams of yellow solids, yield 54% with embodiment 5 with 1-.Mp?174-179℃； 1H?NMR(DMSO，500MHz)δ1.37(t，3H)，4.34(q，2H)，8.47(s，2H)，8.49(s，2H)；MS(ESI)m/z?391.3[M+Na] +。
Embodiment 183, two (2-((2-propylene-1-yl) thiocarbamoyl)) the diazanyl methene bases of 5-)-4-hydroxy-benzoic acid ethyl ester (Compound I A-15) preparation
Figure B200910047268XD0000221
Except (the 1H-benzimidazolyl-2 radicals-yl) hydrazine replaces to 4-(2-propylene-1-yl)-thiosemicarbazide, and all the other desired raw materials, reagent and preparation method get title compound, 27 milligrams of yellow solids, yield 39% with embodiment 5 with 1-.Mp?250-253℃； 1H?NMR(DMSO，500MHz)δ1.37(t，3H)，4.34(q，2H)，4.32(d，4H)，5.14(m，4H)，5.42(m，2H))，8.26(s，2H)，8.43(s，2H)；MS(ESI)m/z?449.2[M] +。
Embodiment 193, two (2-(benzo [1,3] dioxolane-5-formyl radical) diazanyl methene the base)-4-hydroxy-benzoic acid ethyl ester (Compound I of 5- A-16) preparation
Except (the 1H-benzimidazolyl-2 radicals-yl) hydrazine replaces to outside benzo [1,3] dioxolane-5-formyl radical hydrazine, and all the other desired raw materials, reagent and preparation method get title compound, 50 milligrams of yellow solids, yield 81% with embodiment 5 with 1-.Mp?253-258℃； 1H?NMR(DMSO，500MHz)δ1.36(t，3H)，4.34(q，2H)，6.15(s，4H)，7.10(d，2H)，7.50(s，2H)，7.58(d，2H)，8.33(s，2H)，8.77(s，2H)；MS(ESI)m/z?569.1[M+Na] +。
Embodiment 203, two (2-(4-isopropyl phenyl) diazanyl methene the base)-4-hydroxy-benzoic acid ethyl ester (Compound I of 5- A-17) preparation
Figure B200910047268XD0000231
Except (the 1H-benzimidazolyl-2 radicals-yl) hydrazine replaces to the 4-sec.-propyl phenylhydrazine, and all the other desired raw materials, reagent and preparation method get title compound, 28 milligrams of yellow solids, yield 51% with embodiment 5 with 1-.Mp?262-264℃； 1H?NMR(DMSO，500MHz)δ1.19(d，12H)，1.36(t，3H)，2.82(m，2H)，4.34(q，2H)，6.95(d，4H)，7.16(d，4H)，8.22(s，2H)，8.36(s，2H)；MS(EI)m/z?486.3[M] +。
Embodiment 212-hydroxyl-3-(2-(benzo [1,3] dioxolane-5-formyl radical)-5-ethoxycarbonyl-phenylformic acid (Compound I A-18) preparation
Except (the 1H-benzimidazolyl-2 radicals-yl) hydrazine replaces to outside benzo [1,3] dioxolane-5-formyl radical hydrazine, and all the other desired raw materials, reagent and preparation method get title compound, 30 milligrams of yellow solids, yield 94% with embodiment 6 with 1-.Mp?190-195℃； 1H?NMR(DMSO，500MHz)δ1.36(t，3H)，4.34(q，2H)，6.14(s，2H)，7.06(d，1H)，7.49(s，1H)，7.57(d，1H)，8.40(s，1H)，8.54(s，1H)，8.81(s，1H)；MS(EI)m/z?400.1[M] +。
Embodiment 224-nitro-2, the preparation of 6-xylenol (intermediate VII)
Figure B200910047268XD0000241
With 10.0 grams 2, the 6-xylenol is dissolved in 100 milliliters of Glacial acetic acid, under 0 ℃ of the temperature control, splash into the mixing solutions (V/V=1/3) of 16 milliliters of concentrated nitric acids and Glacial acetic acid, keep temperature of reaction to continue to stir 4 hours, in reaction solution impouring frozen water, suction filtration, washing solid, drying, obtain 3.0 gram 4-nitros-2,6-xylenol (intermediate VII).Be red solid, yield 22%.Mp?172-173℃； 1H?NMR(CDCl 3，500MHz)δ2.32(s，6H)，7.96(s，2H)。
Embodiment 232-(the diazanyl methene base-4-nitro-6-methylphenol (Compound I of 1H-benzimidazolyl-2 radicals-yl) B-1) preparation
Figure B200910047268XD0000242
Except with 4-hydroxyl-3,5-mesitylenic acid ethyl ester (intermediate II) replaces to 4-nitro-2, outside the 6-xylenol (intermediate VII), all the other desired raw materials, reagent and preparation method get title compound with embodiment 2,3 and 4,10 milligrams of yellow solids, yield 18%.Mp?253-256℃； 1H?NMR(DMSO，500MHz)δ2.35(s，3H)，7.00(q，2H)，7.11(q，2H)，8.05(s，1H)，8.32(s，1H)，8.46(s，1H)；MS(EI)m/z?311.1[M] +。
Embodiment 242-(the diazanyl methene base-6-methylphenol (Compound I of 1H-benzimidazolyl-2 radicals-yl) C-1) preparation
Figure B200910047268XD0000251
Except with 4-hydroxyl-3,5-mesitylenic acid ethyl ester (intermediate II) replaces to 2, and outside the 6-xylenol, all the other desired raw materials, reagent and preparation method be with embodiment 2,3 and 4, title compound, 76 milligrams of yellow solids, yield 40%.Mp＞300℃； 1H?NMR(DMSO，500MHz)δ1.89(s，3H)，6.81(t，1H)，6.90(d，2H)，6.95(d，2H)，7.12(d，1H)，7.23(d，1H)，8.28(s，1H)；MS(EI)m/z?266.1[M] +。
Embodiment 252, two (2-(1H-benzimidazolyl-2 radicals-yl) the diazanyl methene base) phenol (Compound I of 6- C-2) preparation
Figure B200910047268XD0000252
Except with 4-hydroxyl-3,5-mesitylenic acid ethyl ester (intermediate II) replaces to 2, and outside the 6-xylenol, all the other desired raw materials, reagent and preparation method be with embodiment 2,3 and 5, title compound, 70 milligrams of yellow solids, yield 66%.Mp＞300℃； 1H?NMR(DMSO，500MHz)δ6.96(m，4H)，7.12(t，1H)，7.20(m，4H)，7.71(d，2H)，8.40(s，2H)；MS(EI)m/z?410.2[M] +。
Embodiment 26 The compounds of this invention suppress active mensuration to NS2B-NS3 proteolytic enzyme percentage
(1) the proteic expression and purification of NS2B-NS3
The pET19b carrier that will contain NS2B-Gly-NS3pro181 changes in the e. coli bl21 (DE3), microbial culture and protein induced method are except that inducing temperature being changed into 25 ℃, all the other all referring to the document of Liew etc. (Protein.Expr.Purif.2005,41,332-340.).After inducing 5 hours, 4 ℃, 5000rpm received bacterium in centrifugal 10 minutes.Use the broken bacterium of BugBusterTM high-throughput albumen extraction agent of Novagen company.4 ℃, centrifugal 20 minutes of 8000g collects supernatant liquor, passes through Ni with reference to the document of Liew etc. 2+Affinity chromatography single step purification target protein.Use elution buffer (20mM Tris-HCl, pH 7.9 for 1M imidazoles, 50mM NaCl) with the last wash-out of target protein, and target protein is dialysed to 5mMTris-C1pH 8.0 solution, purity of protein is by the SDS-PAGE electrophoresis detection.Use the endogenous polyclonal antibody of rabbit further to determine target protein NS3pro by immunoblotting.Use immunodetection can detect target protein by densitometric scan (Bio-Rad GS-800 optical density(OD) imager, the U.S.).
(2) The compounds of this invention is to the mensuration of FP-2 enzyme inhibition activity
Damping fluid (damping fluid to 50 μ L, 50mM Tris-C1,0.1%Triton X-100,30% glycerine, pH 9.0) add NS2B-NS3 albumen (final concentration 0.01 μ M) in the system and be dissolved in the testing compound solution of DMSO, hatch the substrate (ultimate density 10 μ M) of adding Dabcyl-KQRRGRIE-Edans behind the 10min under the room temperature.Under different concns compound existence condition, the speed of response that NS2B-NS3 proteolytic enzyme enzyme is cut fluorogenic substrate compares with the speed of response of the 1%DMSO that does not add compound inhibitor, calculates the inhibiting rate under each concentration.The enzymic activity that promptly contains under the 1%DMSO condition is 100%, and suppressing percentage is that 100% relative reactivity that deducts enzyme under the different concns inhibitor multiply by 100%.Enzymatic reaction kinetics is surveyed 60min, excitation wavelength 340nm continuously in Infinite M200 (Tecan) spectrophotofluorometer; Emission wavelength 480nm, average power speed is expressed with RFU/min, calculates testing compound percent inhibition under 100 μ M with following formula,
Calculation formula is: (contrast class value one experiment class value)/contrast class value * 100%
(3) compound activity test result
The polysubstituted single or two phenyl hydrazones compound of table 1 is to NS2B-NS3 inhibiting rate data
Figure B200910047268XD0000271
As can be seen from Table 1, the polysubstituted single or two phenyl hydrazones compound major part with general structure (1) of the present invention has stronger NS2B-NS3 protease inhibiting activity, illustrates that compound of the present invention is the NS2B-NS3 proteinase inhibitor.
Embodiment 27 part of compounds of the present invention are to NS2B-NS3 proteolytic enzyme half effective inhibition concentration (IC 50) mensuration
Choose 100 μ M inhibiting rates and survey IC at the compound more than 70% 50, select the suitable compound concentration gradient, experimental technique and system such as embodiment 26.Enzymatic reaction kinetics is tested in the TECAN spectrophotofluorometer, and average power speed is expressed with RFU/min.Under different concns compound existence condition, the speed of response that NS2B-NS3 proteolytic enzyme enzyme is cut fluorogenic substrate compares with the speed of response that does not add compound inhibitor 1%DMSO, calculates the inhibiting rate under each concentration.The enzymic activity that promptly contains under the 1%DMSO condition is 100%, and suppressing percentage is that 100% relative reactivity that deducts enzyme under the different concns inhibitor multiply by 100%.According to Logistic formula IC 50Value calculates the IC that testing compound suppresses the NS2B-NS3 protease activity in GraphPad Prism5 50Value.Formula is as follows:
A ( I ) / A 0 = 1 - 1 1 + ( I / IC 50 ) p
In the above-mentioned formula, A 0Enzymic activity when referring to add 1%DMSO, A (I) refers to the enzymic activity of compound under I concentration, and I refers to the concentration of testing compound, and p refers to the μ factor.
The polysubstituted single or two phenyl hydrazones compound of table 2 is lived to the NS2B-NS3 enzyme and is suppressed IC 50Value
As can be seen from Table 2, the polysubstituted single or two phenyl hydrazones compound of part with general structure (1) of the present invention has the NS2B-NS3 protease inhibiting activity of little level of rubbing, and illustrates that compound of the present invention is the NS2B-NS3 proteinase inhibitor.
Compound I A-2, I A-5, I A-7, I A-9, I A-14, I A-16 and I C-2 couples of NS2B-NS3 enzyme amount effect curve figure that suppress alive see Fig. 1～Fig. 7.By amount effect curve figure as can be seen, these 7 inhibitor have concentration dependent preferably to the inhibition activity of NS2B-NS3 enzyme.
The possibility of utilizing on the industry
The advantage such as the preparation method of polysubstituted single or two phenyl hydrazones compound of the present invention has that reaction condition gentleness, abundant raw material are easy to get, operation and post processing are simple, and toxicity of compound of the present invention is very low.
Polysubstituted single or two phenyl hydrazones compound of the present invention suppresses all to have shown positive result in the test in the computer virtual screening and at NS2B-NS3 protease. Should demonstrate,prove its pharmacological mechanism. Therefore, compound of the present invention can be used for preparing the NS2B-NS3 enzyme inhibitor and develops into the anti-dengue virus medicine.

Claims (10)

1. a class polysubstituted single or two phenyl hydrazones compound as the formula (1),
Figure F200910047268XC0000011
Wherein:
R 1Be selected from H ,-NO 2With the C1-C4 alkoxy carbonyl;
R 2Be selected from H and-OH;
R 3Be selected from the saturated or unsaturated C1-C10 alkyl, ArC (O) of straight or branched-, Ar-,
Figure F200910047268XC0000012
Figure F200910047268XC0000013
With
Figure F200910047268XC0000014
Perhaps R 3Form 1,3-dihydro-1,3-dioxo-2H-pseudoindoyl with the N that is connected;
R 4Be selected from the C1-C4 alkyl ,-COOH and
Figure F200910047268XC0000015
R 5Be selected from the saturated or unsaturated C1-C4 alkyl and the Ar-of H, straight or branched;
Ar is selected from replacement or unsubstituted phenyl and contains 1～3 and is selected from N; the heteroatomic 5-7 membered aromatic heterocycle base of O and S; and described 5-7 membered aromatic heterocycle base is not necessarily by phenyl and close, and substituting group can be 1～4 and be independently selected from halogen; C1-C6 straight or branched alkyl; hydroxyl; the C1-C4 alkoxyl group; the unsaturated-oxyl of C1-C4; carboxyl; ester group; C1-C6 carboxyl alkoxyl group; C1-C6 ester group alkoxyl group; the C1-C6 carboxyalkyl; C1-C6 ester group alkyl; cyano group; nitro; amino; methylol; trifluoromethyl; trifluoromethoxy; sulfydryl; sulfamyl; the group of dioxolane and C1-C4 acyl group.
2. polysubstituted single or two phenyl hydrazones compound as the formula (1) according to claim 1 is characterized in that, wherein:
R 1Be selected from H ,-NO 2With-COOC 2H 5
R 2Be selected from H and-OH;
R 3Be selected from the saturated or unsaturated C1-C4 alkyl, ArC (O) of straight or branched-, Ar-,
Figure F200910047268XC0000021
Figure F200910047268XC0000022
With
Figure F200910047268XC0000023
Perhaps R 3Form 1,3-dihydro-1,3-dioxo-2H-isoindole-2-base with the N that is connected;
R 4Be selected from-CH 3,-COOH and
R 5Be selected from H ,-CH 3,-C 2H 5,-CH 2-CH=CH 2And Ar-;
Ar is selected from and replaces or unsubstituted phenyl, benzimidazolyl-and pyridyl, and substituting group can be 1～4 group that is independently selected from C1-C6 straight or branched alkyl, C1-C4 alkoxyl group, nitro, dioxolane and sulfamyl.
3. polysubstituted single or two phenyl hydrazones compound as the formula (1) according to claim 2 is characterized in that, is Compound I A, wherein:
R 1Be selected from-COOC 2H 5
R 2Be selected from-OH;
R 3, R 4And R 5Definition identical with claim 2.
4. polysubstituted single or two phenyl hydrazones compound as the formula (1) according to claim 2 is characterized in that, is Compound I B, wherein:
R 1Be selected from-NO 2
R 2Be selected from-OH;
R 4Be selected from-CH 3
R 3And R 5Definition identical with claim 2.
5. polysubstituted single or two phenyl hydrazones compound as the formula (1) according to claim 2 is characterized in that, is Compound I C, wherein:
R 1Be selected from H;
R 2Be selected from-OH;
R 4Be selected from-CH 3With
Figure F200910047268XC0000031
R 3And R 5Definition identical with claim 2.
6. polysubstituted single or two phenyl hydrazones compound as the formula (1) according to claim 2 is characterized in that, is specially:
Figure F200910047268XC0000032
Figure F200910047268XC0000041
Figure F200910047268XC0000051
Figure F200910047268XC0000061
7. the preparation method of the described polysubstituted single or two phenyl hydrazones compound as the formula (1) of claim 1 is characterized in that, described preparation method is selected from any one in following three kinds of methods:
Method one:
1) with 4-hydroxyl-3, the 5-mesitylenic acid is put in the dehydrated alcohol, adds an amount of acid catalyst, and in 60-80 ℃ of stirring 10-20 hour, steaming desolventized, and obtains intermediate II 4-hydroxyl-3,5-mesitylenic acid ethyl ester;
2) intermediate II and acetic anhydride are mixed in 120-140 ℃ stirred 10-20 hour, steaming desolventizes, and obtains intermediate III 4-acetoxy-3,5-mesitylenic acid ethyl ester;
3) intermediate III is joined in an amount of isopyknic Glacial acetic acid and acetic anhydride mixed solution, under temperature control 0-5 ℃, drip an amount of vitriol oil, add excessive chromium trioxide afterwards again, about 2-4 of time spent hour in batches; Keep temperature of reaction to continue to stir 24-48 hour, in reaction solution impouring frozen water, add Sodium Metabisulfite cancellation reaction, stirring at room 15-30 minute; Use organic solvent extraction, drying, steaming desolventizes; Add an amount of ethanol and the catalytic amount vitriol oil to resistates, backflow 3-5 hour; Steaming desolventizes, residual solid is separated through silica gel column chromatography, by the ascending order of polarity, obtain intermediate compound IV 3-formyl radical-4-hydroxy-5-methyl yl benzoic acid ethyl ester, intermediate V 3 successively, 5-diformyl-4-hydroxy-benzoic acid ethyl ester and intermediate VI 2-hydroxyl-3-formyl radical-5-ethoxycarbonyl-phenylformic acid;
4) with three intermediate compound IV, V, VI respectively with R 3The hydrazine that replaces is put in the dehydrated alcohol, adds the Glacial acetic acid of catalytic amount, in 60-80 ℃ of stirring 2-5 hour, and suction filtration, ethanol is washed, and obtains three Compound I respectively A, promptly 3-replaces diazanyl methene base-4-hydroxy-5-methyl yl benzoic acid ethyl ester, 3, and two (replacing diazanyl methene the base)-4-hydroxy-benzoic acid ethyl esters of 5-and 2-hydroxyl-3-replace diazanyl methene base-5-ethoxycarbonyl-phenylformic acid;
Wherein:
R 1Be selected from-COOC 2H 5
R 2Be selected from-OH;
R 3, R 4And R 5Definition identical with claim 1;
Method two:
1) with 2, the 6-xylenol is dissolved in an amount of Glacial acetic acid, temperature control 0-5 ℃, splashes into the mixing solutions of concentrated nitric acid and Glacial acetic acid, keep temperature of reaction to continue to stir 2-5 hour, in reaction solution impouring frozen water, suction filtration, washing solid, drying obtains intermediate VII 4-nitro-2, the 6-xylenol;
2) with intermediate VII with the 2nd in aforesaid method one synthetic) similar method of step prepares intermediate VIII O-(4-nitro-2,6-3,5-dimethylphenyl) acetic ester;
3) with the 3rd in aforesaid method one synthetic), 4) go on foot similar method, prepare Compound I B, promptly 2-replaces diazanyl methene base-4-nitro-6-methyl-phenol;
Wherein:
R 1Be selected from-NO 2
R 2Be selected from-OH;
R 4Be selected from-CH 3
R 3And R 5Definition identical with claim 1;
With,
Method three:
1) with 2, the 6-xylenol is with the 2nd in aforesaid method one synthetic) similar method of step prepares intermediate X O-(2, the 6-3,5-dimethylphenyl) acetic ester;
2) with the 3rd in aforesaid method one synthetic), 4) go on foot similar method, prepare Compound I C, promptly 2-replaces diazanyl methene base-6-methyl-phenol and 2, two (replacing diazanyl methene the base)-phenol of 6-;
Wherein:
R 1Be selected from H;
R 2Be selected from-OH;
R 4Be selected from-CH 3With
R 3And R 5Definition identical with claim 1.
8. a pharmaceutical composition is characterized in that, this pharmaceutical composition comprises described polysubstituted single or two phenyl hydrazones compound and the pharmaceutically acceptable carrier as the formula (1) of one or more claims 1 for the treatment of significant quantity.
9. the purposes of the described polysubstituted single or two phenyl hydrazones compound as the formula (1) of claim 1 in the medicine of preparation dengue virus NS2B-NS3 proteinase inhibitor.
10. the purposes of the described polysubstituted single or two phenyl hydrazones compound as the formula (1) of claim 1 in the medicine that comprises singapore hemorrhagic fever, dengue hemorrhagic fever and dengue shock syndrome that preparation prevention or treatment dengue virus cause.
CN200910047268XA 2009-03-09 2009-03-09 Substituted phenylhydrazones compounds, preparation method and application thereof Expired - Fee Related CN101830854B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN200910047268XA CN101830854B (en) 2009-03-09 2009-03-09 Substituted phenylhydrazones compounds, preparation method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN200910047268XA CN101830854B (en) 2009-03-09 2009-03-09 Substituted phenylhydrazones compounds, preparation method and application thereof

Publications (2)

Publication Number Publication Date
CN101830854A true CN101830854A (en) 2010-09-15
CN101830854B CN101830854B (en) 2012-01-11

Family

ID=42715093

Family Applications (1)

Application Number Title Priority Date Filing Date
CN200910047268XA Expired - Fee Related CN101830854B (en) 2009-03-09 2009-03-09 Substituted phenylhydrazones compounds, preparation method and application thereof

Country Status (1)

Country Link
CN (1) CN101830854B (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102731394A (en) * 2011-04-12 2012-10-17 中国科学院上海药物研究所 Substituted quinoline-2-formaldehyde-phenylhydrazone derivative, and preparation method and application thereof
CN110229081A (en) * 2019-07-01 2019-09-13 长沙理工大学 2,4- dinitrobenzene hydazone derivative and the preparation method and application thereof

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102731394A (en) * 2011-04-12 2012-10-17 中国科学院上海药物研究所 Substituted quinoline-2-formaldehyde-phenylhydrazone derivative, and preparation method and application thereof
CN110229081A (en) * 2019-07-01 2019-09-13 长沙理工大学 2,4- dinitrobenzene hydazone derivative and the preparation method and application thereof
CN110229081B (en) * 2019-07-01 2022-02-08 长沙理工大学 2, 4-dinitrophenylhydrazone derivative and preparation method and application thereof

Also Published As

Publication number Publication date
CN101830854B (en) 2012-01-11

Similar Documents

Publication Publication Date Title
Brecher et al. The flavivirus protease as a target for drug discovery
Ottanà et al. Identification of 5-arylidene-4-thiazolidinone derivatives endowed with dual activity as aldose reductase inhibitors and antioxidant agents for the treatment of diabetic complications
Wang et al. A small-molecule dengue virus entry inhibitor
Hermann et al. Shear stress inhibits H2O2-induced apoptosis of human endothelial cells by modulation of the glutathione redox cycle and nitric oxide synthase
Kiat et al. Inhibitory activity of cyclohexenyl chalcone derivatives and flavonoids of fingerroot, Boesenbergia rotunda (L.), towards dengue-2 virus NS3 protease
Vasiljeva et al. Emerging roles of cysteine cathepsins in disease and their potential as drug targets
Mayhoub et al. Design, synthesis, and biological evaluation of thiazoles targeting flavivirus envelope proteins
Shiryaev et al. Cleavage targets and the D-arginine-based inhibitors of the West Nile virus NS3 processing proteinase
Lima et al. Dual inhibitors of the dengue and West Nile virus NS2B–NS3 proteases: Synthesis, biological evaluation and docking studies of novel peptide-hybrids
Sidique et al. Structure–activity relationship and improved hydrolytic stability of pyrazole derivatives that are allosteric inhibitors of West Nile Virus NS2B-NS3 proteinase
Oliveira et al. NS3 and NS5 proteins: important targets for anti-dengue drug design
Wang et al. Natural products as a gold mine for selective matrix metalloproteinases inhibitors
Pelliccia et al. Inhibition of dengue virus replication by novel inhibitors of RNA-dependent RNA polymerase and protease activities
Srivarangkul et al. A novel flavanone derivative inhibits dengue virus fusion and infectivity
Hou et al. Design, synthesis and biological evaluation of carbohydrate-based sulphonamide derivatives as topical antiglaucoma agents through selective inhibition of carbonic anhydrase II
CN102603695A (en) Amino acid-fluorophore compound and application thereof
Tseng et al. Synthesis, antiproliferative and anti-dengue virus evaluations of 2-aroyl-3-arylquinoline derivatives
CN101830854B (en) Substituted phenylhydrazones compounds, preparation method and application thereof
Samanta et al. Synthesis and in vitro Evaluation of West Nile Virus Protease Inhibitors Based on the 2‐{6‐[2‐(5‐Phenyl‐4H‐{1, 2, 4] triazol‐3‐ylsulfanyl) acetylamino] benzothiazol‐2‐ylsulfanyl} acetamide Scaffold
Meguellati et al. New pseudodimeric aurones as palm pocket inhibitors of Hepatitis C virus RNA-dependent RNA polymerase
Barros et al. Pseudo-peptides derived from isomannide: inhibitors of serine proteases
del Rosario García-Lozano et al. Piperazine-derived small molecules as potential Flaviviridae NS3 protease inhibitors. In vitro antiviral activity evaluation against Zika and Dengue viruses
CN1990479A (en) 3-alkoxy substituted-2, 5, 7-tri-substituted benzopyran-4-ketones compounds, preparing method thereof and pharmaceutical compounds containing same
CN102731394A (en) Substituted quinoline-2-formaldehyde-phenylhydrazone derivative, and preparation method and application thereof
CN110526843A (en) Inhibit calcium/calmodulin dependent protein kinase II compound and its application

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
C17 Cessation of patent right
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20120111

Termination date: 20130309