CN103739617A - Isoxazole norcantharidin derivative, and preparation method and application thereof - Google Patents
Isoxazole norcantharidin derivative, and preparation method and application thereof Download PDFInfo
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- CN103739617A CN103739617A CN201310573156.4A CN201310573156A CN103739617A CN 103739617 A CN103739617 A CN 103739617A CN 201310573156 A CN201310573156 A CN 201310573156A CN 103739617 A CN103739617 A CN 103739617A
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- isoxazole
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- chromone
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D498/22—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains four or more hetero rings
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Abstract
The invention discloses an isoxazole norcantharidin derivative, and a preparation method and an application thereof. The preparation method comprises the following steps: introducing isoxazole rings to the C5 position and the C6 position in a norcantharidin structure, and introducing a chromone structure through a reaction with a chromone derivative in order to synthesize six novel isoxazole norcantharidin derivatives containing the chromone structure. The isoxazole norcantharidin derivatives can be applied to synthesize antitumor medicines, and have a wide application prospect.
Description
Technical field
The invention belongs to medical technical field, relate in particular to a kind of novel containing chromone structure isoxazole norcantharidin derivative and preparation method thereof and application.
Background technology
Norcantharidin, chemical name: 7-oxabicyclo [2.2.1] heptane-2,3-dicarboxylic anhydride, CAS:[5442-12-6; 29745-04-8], chemical structural formula is as follows:
Cantharidin is the effective ingredient of research malignant tumor medicine.Modern study proves, it has certain curative effect to primary hepatocarcinoma, and has leukocyte increasing, and the advantage such as Immunosuppression system, therefore, does not have the very high researching value of will using, and causes people's extensive concern.But the toxicity of Cantharidin is larger, synthetic very complicated, recent research shows, in Norcantharidin, lacked 2, two methyl of 3, Norcantharidin has not only kept stronger anti-tumor activity and unique function of increasing leukocyte, and toxicity reduces greatly, has substantially eliminated Cantharidin urinary system telson is swashed to side effect.
Therefore, the synthetic work that be significant relevant with Cantharidin backbone modification is to remove prosposition methyl substituted.This structural modification can not affect Cantharidin antitumour activity and toxicity decreases, and synthesis step is simplified.
Summary of the invention
The object of this invention is to provide a kind of 6-of utilization methyl-chromone oxime transforms in the structure of isoxazole, Norcantharidin is carried out to structure of modification, improve the active novel containing chromone structure isoxazole norcantharidin derivative of Norcantharidin, and provided its preparation method and application.
In order to solve the problems of the technologies described above, the present invention is solved by following technical proposals:
An isoxazole norcantharidin derivative, its general structure is as follows:
Wherein, R
1hydrogen, hydroxyl, methyl, nitro, chlorine or bromine.
The preparation method of above-mentioned isoxazole norcantharidin derivative, comprises the steps:
Step 1): dehydronorcantharidiimide element synthetic: by MALEIC ANHYDRIDE porphyrize, add ether, be stirred to dissolving under room temperature condition, splash into furans, stirring at room 24-48 hour, there is Diels-Alder with MALEIC ANHYDRIDE and react in furans, makes dehydronorcantharidiimide element after filtration under diminished pressure;
Step 2): N-substituted dehydronorcantharidiimide imide synthetic: dehydronorcantharidiimide element is dissolved in acetone solvent, under agitation drips the acetone soln of aniline, react and add manganese acetate, triethylamine and acetic anhydride after 1 hour, at room temperature react 8 hours; Reacted precipitation is dissolved in to N after vacuum-drying, in dinethylformamide, in ice-water bath, with dicyclohexylcarbodiimide stirring reaction 10 hours, filtrate is placed in to frozen water and separates out precipitation, then obtain N-substituted dehydronorcantharidiimide imide by sedimentation and filtration and with recrystallization;
Step 3): import chromone structure: N-substituted dehydronorcantharidiimide imide and 6-methyl-chromone oxime are mixed in ethanol, add chloramine-T, reflux 12 hours, carry out 1,3-Dipolar Cycloaddition, use methyl alcohol recrystallize, vacuum-drying obtains product.
As preferably, the preparation method of described 6-methyl-chromone oxime is as follows: in Erlenmeyer flask, add oxammonium hydrochloride and water, magnetic agitation is molten clear to oxammonium hydrochloride; Under agitation, taking 6-methyl-chromone and ethanol pours in Erlenmeyer flask, vigorous stirring, after question response is complete, it is neutral with sodium carbonate solution, regulating the pH value of reaction solution, placement is cooled to room temperature, produce a large amount of yellow mercury oxides, put into after refrigerator overnight filtration under diminished pressure, drying at room temperature, obtains the yellow crystals of needle-like.
As preferably, described step 2) in, the temperature of ice-water bath is down to 0 ℃.
As preferably, described step 2) recrystallization in adopts methyl alcohol.
Above-mentioned isoxazole norcantharidin derivative is for the synthesis of antitumor drug.
The reaction the present invention relates to is as follows:
R
1hydrogen, hydroxyl, methyl, nitro, muriate or bromine.
4a R
1hydrogen; 4b R
1it is hydroxyl; 4c R
1it is methyl; 4d R
1it is nitro; 4e R
1chlorine; 4f R
1it is bromine.
Find after deliberation, the oxygen in Norcantharidin five-ring can be by nitrogen or sulfur, and some substituting groups can be substituted on nitrogen and sulphur.Simultaneously at C
5and C
6that upper replacement can also change pharmacologically active.We draw following experimental data through research work for this reason:
Based on above experimental data, we use 6-methyl-chromone oxime to transform in the structure of isoxazole in the present invention, and Norcantharidin is carried out to structure of modification, have improved the activity of Norcantharidin.
The present invention, owing to having adopted above technical scheme, has significant technique effect:
The inventive method is used the structure of 6-methyl-chromone oxime Dui isoxazole to transform, and Norcantharidin is carried out to structure of modification, has improved the anti-tumor activity of Norcantharidin.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in further detail:
The general structure of the raw material of the synthetic norcantharidin derivative of the inventive method is as follows:
Wherein, described compound 5 is that 6-methyl-chromone reacts the 6-methyl-chromone oxime generating with oxammonium hydrochloride.
6-methyl-chromone oxime preparation: in 250ml Erlenmeyer flask, add 35.6g(0.5mol) oxammonium hydrochloride (M=69.5) and 90mlH
2o, magnetic agitation is molten clear to oxammonium hydrochloride.Under agitation, take (0.4mol) 6-methyl-chromone and 50ml dehydrated alcohol and pour in 250ml Erlenmeyer flask, vigorous stirring 2h.After question response is complete, use 20%Na
2cO
3the about 300ml of solution, the pH value that regulates reaction solution be neutrality, placement is cooled to room temperature, produces a large amount of yellow mercury oxides, put into after refrigerator overnight, filtration under diminished pressure, drying at room temperature, obtains the yellow crystals of needle-like.
Embodiment 1
Synthesizing of dehydronorcantharidiimide element: add successively 15mL ethyl ester in 100mL Erlenmeyer flask, the Powdered MALEIC ANHYDRIDE of 4.00g (40mmol).After MALEIC ANHYDRIDE dissolving, under stirring, add 2.76g(40.5mmol) furans.And then place 24-48 hour under room temperature, the product obtaining after reacting completely is carried out to filtration under diminished pressure and obtain Norcantharidin.
Synthesizing of N-substituted dehydronorcantharidiimide imide: get 3.32g (20mmol) Norcantharidin and be dissolved in 30mL acetone, acetone soln and compound 2 are joined in reaction Erlenmeyer flask, have as seen a large amount of precipitations to generate.React at normal temperatures after 8 hours, by sedimentation and filtration, out final vacuum is dry, is dissolved in 20mL dimethyl formamide, is cooled to 0 ℃.Be placed in ice-water bath, add 3.09g (15mmol) dicyclohexylcarbodiimide, stirring reaction 10 hours.Then cooling, filter, filtrate is poured in 50mL frozen water, separate out solid.Filtered, washing, finally obtains product 3a-3f by recrystallizing methanol.
Import chromone structure: 1mmolN-substituted dehydronorcantharidiimide imide and 1.1mmol6-methyl-chromone oxime, in 20mL ethanol, then are added to 1.2mmol chloramine-T, reflux 12 hours.By sedimentation and filtration, by recrystallizing methanol, after vacuum-drying, obtain 4a, 4b, 4c, 4d, these 6 products of 4e, 4f.
Specific experiment result is as follows:
4,8-epoxy-5,7-(1H, 6H)-diketone-6-(4-phenyl)-3a, 4,4a, 7a, 8,8a-, six hydrogen-3-(6-methyl-4H-chromone-4-ketone)-pyrroles [3,4-f]-1,2-Ben Bing isoxazole (4a)
m.p.104.8-105.3℃
1H?NHR(DMSO)δ:2.44(s,3H,CH3),6.62-7.72(m,7H,Ar-H),7.41(s,1H,O=C-H),3.55(s,H,C5-H),2.38(s,1H,C6-H),5.26(s,2H,C4-H),4.83(s,1H,C1-H,),3.10(s,2H,C2-H,C3-H).
IR3420(N-C=O),3258(ArH),1713(C=O),1618(C=N),1287(C-O-C)cm
-1
Anal.calcd.for?C25H18N2O6.C,67.68;H,4.10;Cl,6.42;N,6.33.
4,8-epoxy-5,7-(1H, 6H)-diketone-6-(4-hydroxyphenyl)-3a, 4,4a, 7a, 8,8a-, six hydrogen-3-(6-methyl-4H-chromone-4-ketone)-pyrroles [3,4-f]-1,2-Ben Bing isoxazole (4b)
m.p.110.4-111.5℃
1H?NHR(DMSO)δ:2.44(s,6H,CH3),8.09-7.07(m,7H,Ar-H),7.14(s,1H,O=C-H),2.35-2.38(d,J=12HZ,H,C5-H),2.54(s,1H,C6-H),5.25(s,1H,O-H),4.83(s,1H,C4-H),3.07(s,1H,C1-H),3.52(s,2H,C2-H,C3-H).
IR:3354(N-C=O),3260(ArH),1557(C=N),1715(C=O),1198(C-O-C)cm
-1
Anal.calcd.for?C25H18N2O7.C,65.50;H,3.94;N,6.12.
4,8-epoxy-5,7-(1H, 6H)-diketone-6-(4-p-tolyl)-3a, 4,4a, 7a, 8,8a-, six hydrogen-3-(6-methyl-4H-chromone-4-ketone)-pyrroles [3,4-f]-1,2-Ben Bing isoxazole (4c)
m.p.108.9-110.0℃
1H-NHR(DMSO)δ:2.44(S,6H,CH3),8.09-7.07(m,7H,Ar-H),7.14(s,1H,O=C-H),2.35-2.38(d,J=12HZ,H,C5-H),2.54(s,1H,C6-H),5.25(s,1H,C4-H),3.07(s,1H,C1-H),3.52(s,2H,C2-H,C3-H).
IR3357(N-C=O),3260(ArH),1714(C=N),1567(C=O),1189(C-O-C)cm
-1
Anal.calcd.for?C26H20N2O6.C,68.40;H,4.43;N,6.14.
4,8-epoxy-5,7-(1H, 6H)-diketone-6-(4-nitrophenyl)-3a, 4,4a, 7a, 8,8a-, six hydrogen-3-(6-methyl-4H-chromone-4-ketone)-pyrroles [3,4-f]-1,2-Ben Bing isoxazole (4d)
m.p.98.5-99.5℃
1H-NHR(DMSO)δ:2.44(s,3H,CH3),6.63-8.39(m,7H,Ar-H),8.08(s,1H,O=C-H),5.29(s,H,C5-H),2.38(s,1H,C6-H),6.46(s,1H,C4-H),3.15(s,1H,C1-H),3.52(s,2H,C2-H,C3-H).
IR3413(N-C=O),3230(ArH),1596(C=N),1720(C=O),1259(C-O-C)cm
-1
Anal.calcd.for?C25H17N3O8.C,61.60;H,3.54;N,8.61.
4,8-epoxy-5,7-(1H, 6H)-diketone-6-(4-chloro-phenyl-)-3a, 4,4a, 7a, 8,8a-, six hydrogen-3-(6-methyl-4H-chromone-4-ketone)-pyrroles [3,4-f]-1,2-Ben Bing isoxazole (4e)
m.p.93.8-95.1℃
1H-NHR(DMSO)δ:2.44(s,3H,CH3),7.16-7.73(m,7H,Ar-H),7.16(s,1H,O=C-H),2.38(s,H,C6-H),3.10(s,1H,C5-H),3.46(s,2H,C4-H,C1-H),2.52(s,2H,C2-H),2.53(s,2H,C3-H)
IR3227(N-C=O),3086(ArH),1557(C=N),1715(C=O),1160(C-O-C)cm
-1
Anal.calcd.for?C25H17ClN2O6.C,62.95;H,3.60;N,5.90.
4,8-epoxy-5,7-(1H, 6H)-diketone-6-(4-bromophenyl)-3a, 4,4a, 7a, 8,8a-, six hydrogen-3-(6-methyl-4H-chromone-4-ketone)-pyrroles [3,4-f]-1,2-Ben Bing isoxazole (4f)
m.p.202.0-203.5℃
1H?NHR(DMSO)δ:2.44(s,3H,CH3),6.19-7.73(m,7H,Ar-H),7.37(s,1H,O=C-H),3.55(s,H,C5-H),2.38(s,1H,C6-H),5.26(s,1H,C4-H,),4.83(s,1H,C1-H,),3.10(s,2H,C2-H,C3-H).
IR3461(N-C=O),3089(ArH),1488(C=N),1716(C=O),1181(C-O-C)cm
-1
Anal.calcd.for?C25H17BrN2O6.C,57.60;H,3.27;N,5.36.
The antitumor cytolytic activity result of norcantharidin derivative
Mtt assay is measured the In-vitro Inhibitory Effect of three series compounds to different knurl strains:
Compound is dissolved with DMSO, dilution, tumour cell Bel-7402(human liver cancer cell), KB(cancer cell of oral cavity), SGC7901(stomach cancer cell), HO8901(ovarian cancer cell), HL-60(leukemia cell), ECA109(colon-cancer cell), on 96 orifice plates, plant into 4000/200 μ L/ holes, every hole adds compound 2 μ L, final concentration is 12.0 μ M, 6.0 μ M, 3.0 μ M, 1.5 μ M, jointly in 37 ℃, 5%CO
2in cell culture incubator, hatch 72 hours, take DMSO(1%) be blank.After 72 hours, adding final concentration is the MTT of 0.25mg/mL, is placed in 37 ℃, 5%CO
2in cell culture incubator 4 hours, blot afterwards solvent, every hole adds 100 μ l DMSO, and with enzyme linked immunological instrument, in 570nm place mensuration absorbancy (OD value), the data obtained is used for calculating IC
50value.Select and suppress active high compound, the impact of the compound effects asynchronism(-nization) under mensuration different concns on human tumor cells cycle and apoptosis.
The test-compound of different concns carries out scalping with 96 orifice plates, according to the inhibiting rate of gained, calculates IC
50value, the results are shown in following table.
Table3-1IC
50values?of?of?the?norcantharidin?derivatives?against?six?tumor?cell?lines
In a word, the foregoing is only preferred embodiment of the present invention, all equalizations of doing according to the present patent application the scope of the claims change and modify, and all should belong to the covering scope of patent of the present invention.
Claims (6)
1. an isoxazole norcantharidin derivative, its general structure is as follows:
Wherein, R
1hydrogen, hydroxyl, methyl, nitro, chlorine or bromine.
2. the preparation method of isoxazole norcantharidin derivative according to claim 1, is characterized in that, comprises the steps:
Step 1): dehydronorcantharidiimide element synthetic: by MALEIC ANHYDRIDE porphyrize, add ether, be stirred to dissolving under room temperature condition, splash into furans, stirring at room 24-48 hour, there is Diels-Alder with MALEIC ANHYDRIDE and react in furans, makes dehydronorcantharidiimide element after filtration under diminished pressure;
Step 2): N-substituted dehydronorcantharidiimide imide synthetic: dehydronorcantharidiimide element is dissolved in acetone solvent, under agitation drips the acetone soln of aniline, react and add manganese acetate, triethylamine and acetic anhydride after 1 hour, at room temperature react 8 hours; Reacted precipitation is dissolved in to N after vacuum-drying, in dinethylformamide, in ice-water bath, with dicyclohexylcarbodiimide stirring reaction 10 hours, filtrate is placed in to frozen water and separates out precipitation, then obtain N-substituted dehydronorcantharidiimide imide by sedimentation and filtration and with recrystallization;
Step 3): import chromone structure: N-substituted dehydronorcantharidiimide imide and 6-methyl-chromone oxime are mixed in ethanol, add chloramine-T, reflux 12 hours, carry out 1,3-Dipolar Cycloaddition, use methyl alcohol recrystallize, vacuum-drying obtains product.
3. the preparation method of isoxazole norcantharidin derivative according to claim 2, is characterized in that: the preparation method of described 6-methyl-chromone oxime is as follows: in Erlenmeyer flask, add oxammonium hydrochloride and water, magnetic agitation is molten clear to oxammonium hydrochloride; Under agitation, taking 6-methyl-chromone and ethanol pours in Erlenmeyer flask, vigorous stirring, after question response is complete, it is neutral with sodium carbonate solution, regulating the pH value of reaction solution, placement is cooled to room temperature, produce a large amount of yellow mercury oxides, put into after refrigerator overnight filtration under diminished pressure, drying at room temperature, obtains the yellow crystals of needle-like.
4. the preparation method of isoxazole norcantharidin derivative according to claim 1, is characterized in that: described step 2), the temperature of ice-water bath is down to 0 ℃.
5. the preparation method of isoxazole norcantharidin derivative according to claim 1, is characterized in that: the recrystallization described step 2) adopts methyl alcohol.
6. according to the arbitrary described isoxazole norcantharidin derivative of claim 1-5, be used for the synthetic of antitumor drug.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002076989A1 (en) * | 2001-03-23 | 2002-10-03 | The University Of Newcastle Research Associates Limited | Protein phosphate inhibitors |
| CN101812066A (en) * | 2010-03-04 | 2010-08-25 | 绍兴文理学院 | Pyrazolo N-substituted dehydronorcantharidin imide derivative as well as synthesis method, activity test method and application thereof |
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2013
- 2013-11-15 CN CN201310573156.4A patent/CN103739617B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002076989A1 (en) * | 2001-03-23 | 2002-10-03 | The University Of Newcastle Research Associates Limited | Protein phosphate inhibitors |
| CN101812066A (en) * | 2010-03-04 | 2010-08-25 | 绍兴文理学院 | Pyrazolo N-substituted dehydronorcantharidin imide derivative as well as synthesis method, activity test method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| LI-PING DENG，等: "1,3-Dipolar Cycloaddition Reaction: Synthesis of Novel 5,6-Dehydronorcantharidin Derivatives of Substituted Aromatic Amines with potential antitumor activities", 《J. HETEROCYCLIC CHEM.》 * |
| 刘方明，等: "1,3-偶极环加成合成新型4,5-二氢-2,5-二氧杂吡咯啉并[4,5-c]异唑衍生物", 《有机化学》 * |
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