CN104804066A - Novel anti-cancer compound, preparation method for anti-cancer compound and application of anti-cancer compound to preparation of anti-cancer drugs - Google Patents

Novel anti-cancer compound, preparation method for anti-cancer compound and application of anti-cancer compound to preparation of anti-cancer drugs Download PDF

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CN104804066A
CN104804066A CN201510219524.4A CN201510219524A CN104804066A CN 104804066 A CN104804066 A CN 104804066A CN 201510219524 A CN201510219524 A CN 201510219524A CN 104804066 A CN104804066 A CN 104804066A
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formula
compound
reaction
compound shown
distilled water
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魏元刚
严晓强
金显友
钟慧
杨永安
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JIANGSU NAIQUE BIOLOGICAL ENGINEERING TECHNOLOGY Co Ltd
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JIANGSU NAIQUE BIOLOGICAL ENGINEERING TECHNOLOGY Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J71/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton is condensed with a heterocyclic ring
    • C07J71/0005—Oxygen-containing hetero ring
    • C07J71/001—Oxiranes

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Abstract

The invention discloses a novel anti-cancer compound with a structure as shown in a formula VIII, a preparation method for the anti-cancer compound and an application of the anti-cancer compound to the preparation of anti-cancer drugs. In the formula, R1 is H, CH3 or CH2CH3. The novel anti-cancer compound has the advantages of better bioactivity, higher selectivity, lower toxicity, shorter residual life and the like.

Description

New type anticancer compound, its preparation method and prepare the purposes of cancer therapy drug
Technical field
The invention belongs to medicinal chemistry art, especially a kind of anticancer compound, its preparation method and prepare the purposes of cancer therapy drug.
Background technology
Through the steroidal saponin constituents that extraction separation and purification obtains from Chinese medicine Caulis Marsdeniae Tenacissimae, its aglycon is by unique steroidal saponin structure of 21 C atomic buildings, is therefore called C 21steroidal saponin.Now there are some researches show that there is C 21the pharmacologically active such as the compound of steroidal saponin parent nucleus has immunomodulatory, relieving asthma, has obtained clinically as the index activeconstituents of medicine clinically and has applied more widely.
Contriver finds after deliberation: have C 21the compound of steroidal saponin parent nucleus has good antineoplastic activity, has good restraining effect to kinds of tumors, as all having good inhibit activities to liver cancer, lung cancer, esophagus cancer etc.At present to this type of C 21the research of steroidal saponin also not deeply, is mainly carried out separation and purification and analyzing and testing research according to its physico-chemical property to it, is carried out structural modification research and carry out pharmacology activity research for its constitutional features for its parent nucleus.
How to make it have better biological activity, higher selectivity, lower toxicity is current important research content.
Summary of the invention
Goal of the invention a: object is to provide a kind of antineoplastic compound, at least to solve the subproblem that prior art exists.Further object is to provide a kind of preparation method of antineoplastic compound.Further object is to provide a kind of antineoplastic compound and is preparing the application in antitumor drug.
Technical scheme: a kind of structure such as formula the compound shown in VIII,
Wherein, R 1for H, CH 3, CH 2cH 3.
Structure such as formula a preparation method for the compound shown in VIII,
Wherein, R 1for H, CH 3, CH 2cH 3.
Comprise the steps:
Step 1: prepare hydrazino-benzoic acid methyl esters, and by structure such as formula the converting compounds shown in I for structure is such as formula the compound shown in II;
Step 2: add diazomethane ether mixed solution, boron trifluoride-ether complex and structure such as formula the compound shown in II in reactor, be obtained by reacting the compound of structure as shown in formula III;
Step 3: add the chemical combination of structure as shown in formula III, the KOH aqueous solution, structure such as formula the compound shown in IV and ethanol in reactor, obtain structure after reaction such as formula the compound shown in V;
Step 4: by structure such as formula the compound dehydrated alcohol shown in V, join in reaction vessel to hydrazino-benzoic acid methyl esters and Glacial acetic acid, be obtained by reacting structure such as formula the compound shown in IV;
Step 5: join in the hydroxylamine solution of activation such as formula the compound shown in IV by structure, is obtained by reacting structure such as formula the compound shown in VII;
Step 6: add structure such as formula the compound shown in VII, methylene dichloride in reactor, and drip boron tribromide, be obtained by reacting target product;
Formula IV ~ VII, R 1for H, CH 3, CH 2cH 3.
In a preferred embodiment, described step 1 is further:
At 0 ± 5 DEG C, add in reactor successively anhydrous methanol, sulfur oxychloride, to hydrazino-benzoic acid, after stirring 30 ~ 40min, continue stirring 3 ~ 5h at normal temperatures, the solid filtering gained uses distilled water, ethanol, distilled water wash successively, dry, and the solid product obtained is dissolved in dehydrated alcohol, recrystallization is purified, and obtains hydrazino-benzoic acid methyl esters.
In room temperature (herein, room temperature refers to 15 DEG C ~ 25 DEG C) stir under, in reactor, add structure successively such as formula the compound shown in I, aqueous hydrochloric acid, dehydrated alcohol, stir after 2 ~ 3h, back flow reaction 7-9h, TLC follows the tracks of reaction, after reaction terminates, filters the solid obtained and uses aqueous hydrochloric acid, distilled water, ethanol, distilled water wash successively, dry, the solid crude product obtained is dissolved in dehydrated alcohol, and recrystallization is purified, and obtains structure such as formula the compound shown in II.
Preferred, structure is dissolved in such as formula the compound 10mmol shown in I the hydrochloric acid 5ml that dehydrated alcohol 50ml. drips 1mol/L.
In a preferred embodiment, described step 2 is further:
Under ice-water bath stirring action, diazomethane ether mixed solution, boron trifluoride-ether complex is added successively in reactor, and the compound of structure as shown in formula III, after reaction 20 ~ 40min, back flow reaction 5 ~ 7h again, TLC follows the tracks of reaction, and after reaction terminates, filtration obtains solid and uses aqueous hydrochloric acid, distilled water, ethanol, distilled water wash successively, dry, the solid product obtained is dissolved in dehydrated alcohol, and recrystallization is purified, and obtains the compound of structure as shown in formula III.
Preferred, in above-mentioned steps, each reactant molar ratio is the compound of structure as shown in formula III: diazomethane: boron tribromide=1:1.2 ~ 1.5:1.5 ~ 2.0.Preferred further, ratio is 1:1.2:1.5,1:1.5:1.5,1:1.2:2.0.
In a preferred embodiment, described step 3 is further:
Under stirring at room temperature, the compound of structure as shown in formula III, the KOH aqueous solution, structure is added successively such as formula the compound shown in IV, ethanol in reactor, after continuing stirring reaction 3 ~ 6h, pH is regulated with hcl acidifying, filter the solid obtained and use distilled water, ethanol, distilled water wash successively, the dry structure that obtains is such as formula the compound shown in V.
Preferred, each reactant molar ratio in above-mentioned steps: structure is such as formula the compound shown in IV: the compound of structure as shown in formula III: boron tribromide=1:1:1 ~ 1.5.
In a preferred embodiment, described step 4 is further:
Successively by structure such as formula the compound shown in V, dehydrated alcohol, join in reactor to hydrazino-benzoic acid methyl esters and Glacial acetic acid, after stirring at room temperature reaction 20 ~ 40min, have fraction solids insoluble; Then back flow reaction 5 ~ 8h, TLC follow the tracks of reaction, after reaction terminates, filter, solid uses aqueous hydrochloric acid, distilled water, ethanol, distilled water wash successively, dry, the solid product obtained is dissolved in dehydrated alcohol, and recrystallization is purified, and obtains structure such as formula the compound shown in VI.
Preferred, each reactant molar ratio: structure is such as formula the compound shown in V: to Hydrazinobenzenesulfonamide: acetic acid=1:1.2 ~ 1.5:0.5.Preferred further, ratio is: 1:1.2:0.5,1:1.5:0.5.
In a preferred embodiment, described step 5 is further:
Hydrochloric acid hydroxyl is added in anhydrous methanol by, sodium methylate, after heated and stirred 20 ~ 40min, filters solid, obtain the hydroxylamine solution activated, add structure wherein such as formula the compound shown in VI, back flow reaction 3 ~ 5h, TLC follows the tracks of reaction, after reaction terminates, filter, the solid distilled water wash obtained, last vacuum-drying, the solid obtained is dissolved in dehydrated alcohol, and recrystallization is purified, and obtains structure such as formula the compound shown in VII.
Preferred, each reactant molar ratio: VI: hydrochloric acid hydroxyl is pressed: sodium methylate=1:4 ~ 5:4 ~ 5.Preferred further, ratio is 1:4:4,1:5:4.
In a preferred embodiment, described step 6 is further:
Under stirring at-20 ± 5 DEG C, in reactor, add structure successively such as formula the compound shown in VII, methylene dichloride, and progressively drip boron tribromide, stir 20 ~ 40min, room temperature continues reaction 8 ~ 12h, and TLC follows the tracks of reaction, after reaction terminates, filter, solid distilled water wash, last vacuum-drying, is dissolved in dehydrated alcohol by the solid obtained, recrystallization is purified, and obtains lenticular target compound.
Preferred, in above-mentioned steps, each reactant molar ratio is: VII: boron tribromide=1:0.5
Present method is transformed for saponin(e aglycon, has effectively integrated hydroxamic acid skeleton, pyrazoline skeleton, assay reproducibility is strong, good stability, and the required condition of experiment reaction is simpler, and experimental situation is gentle, productive rate is better, can produce in a large number in less input situation.
Structure is preparing the application in cancer therapy drug such as formula the compound shown in VIII,
Wherein, R 1for H, CH 3, CH 2cH 3.
A kind of anticancer pharmaceutical composition, comprises structure such as formula the compound shown in VIII and medically acceptable carrier.
Beneficial effect: the present invention has the advantages such as better biological activity, higher selectivity and lower toxicity.There is obvious restraining effect to human breast cancer cell (MCF-7), cervical cancer cell (HeLa), lung carcinoma cell (A549) and liver cancer cell (HepG2), people's renal epithelial cell (293T) shown quite simultaneously or be better than the cytotoxicity of positive control medicine Celecoxib.
Embodiment
The implementation process concrete in conjunction with certain describes the present invention.The structural formula of each reactant, intermediate product and target product is as shown in literary composition:
The present invention is made up of the following step:
Step 1. is at 0 ± 5 DEG C, add in round-bottomed flask successively anhydrous methanol, sulfur oxychloride, to hydrazino-benzoic acid 1, after stirring about 1h, normal temperature continues to stir 3-5h, filter, the solid obtained uses distilled water (3 × 150mL), cold ethanol (3 × 50mL), distilled water (3 × 100mL) to wash successively, dry, the solid crude product obtained is dissolved in dehydrated alcohol recrystallization and obtains carboxylate to hydrazino-benzoic acid methyl esters 2.
Step 2. under stirring at room temperature, adds Tenacissoside A 1 (a kind of C successively in round-bottomed flask 21steroidal saponin) 3, dilute hydrochloric acid, dehydrated alcohol, stir after 1h, back flow reaction 7-9h.TLC follows the tracks of reaction (developping agent V acOEt: V pE=1:2), after reaction terminates, filter, the solid obtained uses dilute hydrochloric acid (3 × 100mL), distilled water (3 × 150mL), cold ethanol (3 × 50mL), distilled water (3 × 100mL) to wash successively, drying, is dissolved in the lenticular C that dehydrated alcohol recrystallization obtains partial reduction by the solid crude product obtained 21steroidal saponin aglycon 4.
Step 3., under ice-water bath stirring action, adds diazomethane ether mixed solution, boron trifluoride-ether complex, C successively in round-bottomed flask 21steroidal saponin aglycon 4, after reaction about 0.5h, then back flow reaction 5-7h.TLC follows the tracks of reaction (developping agent V acOEt: V pE=1:2), after reaction terminates, filter, obtaining solid uses dilute hydrochloric acid (3 × 100mL), distilled water (3 × 150mL), cold ethanol (3 × 50mL), distilled water (3 × 100mL) to wash successively, drying, is dissolved in dehydrated alcohol recrystallization by the solid crude product obtained and obtains lenticular intermediate 5.
Step 4. under stirring at room temperature, intermediate 5, the KOH aqueous solution, the phenyl aldehyde of different substituents, ethanol is added successively in round-bottomed flask, after continuing stirring reaction 4h, dilute hydrochloric acid acidifying regulates pH, filter, the solid obtained uses distilled water (3 × 100mL), cold ethanol (3 × 50mL), distilled water (3 × 100mL) to wash successively, dry intermediate 6-8.
Step 5., successively by different compound 6-8, dehydrated alcohol, join in round-bottomed flask to hydrazino-benzoic acid methyl esters 2, Glacial acetic acid, after stirring at room temperature reaction 1h, still has fraction solids insoluble; Then back flow reaction 5h, TLC follow the tracks of reaction (developping agent V acOEt: V pE=1:2), after reaction terminates, filter, solid uses dilute hydrochloric acid (3 × 100mL), distilled water (3 × 150mL), cold ethanol (3 × 50mL), distilled water (3 × 100mL) to wash successively, drying, is dissolved in dehydrated alcohol recrystallization and obtains lenticular intermediate 9-11 by the solid crude product obtained.
Equimolar hydrochloric acid hydroxyl adds in anhydrous methanol by, sodium methylate by step 6., and after heated and stirred 0.5h, filters solid, obtains the hydroxylamine solution activated, and is adding corresponding crystal intermediate 9-11 wherein, back flow reaction 3-5h.TLC follows the tracks of reaction (developping agent V acOEt: V pE=1:2), after reaction terminates, filter, solid distilled water wash, last vacuum-drying, is dissolved in dehydrated alcohol by the solid obtained, and recrystallization is purified, and obtains lenticular target compound 12-14.
Step 7. adds corresponding intermediate 12-14, methylene dichloride successively, and progressively drips boron tribromide under stirring at-20 ± 5 DEG C in round-bottomed flask, and after stirring about 1h, room temperature continues reaction about 12h.TLC follows the tracks of reaction (developping agent V acOEt: V normal hexane=1:2), after reaction terminates, filter, solid distilled water wash, last vacuum-drying, is dissolved in dehydrated alcohol by the solid obtained, and recrystallization is purified, and obtains lenticular target compound 15-17.
Embodiment one:
4-(3-((3aR, 4aS, 8S, 10aS, 11S, 12S, 12aS)-(8,11,12-trihydroxy-)-(10a, 12a-dimethyl)-(1,2-pentamethylene base)-1,10a-b-epoxy n-tetradecane base)-(4,5-pyrazoline)) preparation of benzoyl hydroxamic acid (compound 18)
Under stirring at-20 DEG C, corresponding intermediate 14 (10.0mmol), methylene dichloride (25mL) that step 6 obtains is added successively in the round-bottomed flask of 50mL, and progressively drip after boron tribromide (5.0mmol) continues stirring reaction 1h, reaction flask is transferred to room temperature, continues reaction 12h.TLC follows the tracks of reaction (developping agent V acOEt: V normal hexane=1:2), after reaction terminates, filter, solid distilled water wash, last vacuum-drying, is dissolved in dehydrated alcohol by the solid obtained, and recrystallization is purified, and obtains lenticular target compound 15.
Obtain white crystal, productive rate 69.3%.m.p.188～189℃； 1H NMR(DMSO-d 6,300MHz)δ:10.69(s,1H,NH),10.50(s,1H,OH),8.00(d,J＝8.1Hz,2H,ArH),7.75(d,J＝7.7Hz,2H,ArH),5.37(s,2H,OH),4.49(s,1H,OH),3.58～3.33(m,5H,CH and CH 2),2.38～2.31(m,2H,CH 2),2.01(t,J＝7.3Hz,1H,CH),1.79～1.21(m,15H,CH and CH 2),1.14(dd,J 1＝7.1Hz,J 2＝7.8Hz,1H,CH),0.89(s,3H,CH 3),0.84(s,3H,CH 3).ESI-MS:546.7[M+H] +.Anal.Calcd forC 28H 39N 3O 5.
Embodiment two:
4-(5-methyl-3-((3aR, 4aS, 8S, 10aS, 11S, 12S, 12aS)-(8,11,12-trihydroxy-)-(10a, 12a-dimethyl)-(1,2-pentamethylene base)-1,10a-b-epoxy n-tetradecane base)-(4,5-pyrazoline)) preparation of benzoyl hydroxamic acid (compound 19)
Preparation method's reference example one.
Obtain white crystal, productive rate 78.6%.m.p.197～198℃； 1H NMR(DMSO-d 6,300MHz)δ:10.69(s,1H,NH),10.50(s,1H,OH),7.97(d,J＝8.2Hz,2H,ArH),7.78(d,J＝7.5Hz,2H,ArH),5.37(s,2H,OH),4.49(s,1H,OH),3.58～3.33(m,3H,CH),2.75～2.71(m,1H,CH),2.38～2.31(m,2H,CH 2),2.01(t,J＝7.3Hz,1H,CH),1.79～1.21(m,18H,CH,CH 2and CH 3),1.14(dd,J 1＝7.1Hz,J 2＝7.8Hz,1H,CH),0.89(s,3H,CH 3),0.83(s,3H,CH 3).ESI-MS:560.7[M+H] +.Anal.Calcd for C 29H 41N 3O 5.
Embodiment three:
4-(5-ethyl-3-((3aR, 4aS, 8S, 10aS, 11S, 12S, 12aS)-(8,11,12-trihydroxy-)-(10a, 12a-dimethyl)-(1,2-pentamethylene base)-1,10a-b-epoxy n-tetradecane base)-(4,5-pyrazoline)) preparation of benzoyl hydroxamic acid (compound 20)
Preparation method's reference example one.
Obtain white crystal, productive rate 82.4%.m.p.202～203℃； 1H NMR(DMSO-d 6,300MHz)δ:10.71(s,1H,NH),10.52(s,1H,OH),8.03(d,J＝8.2Hz,2H,ArH),7.79(d,J＝7.6Hz,2H,ArH),5.35(s,2H,OH),4.49(s,1H,OH),3.58～3.33(m,3H,CH),2.75～2.71(m,1H,CH),2.38～2.29(m,2H,CH 2),2.01(t,J＝7.3Hz,1H,CH),1.79～1.21(m,17H,CH and CH 2),1.14(dd,J 1＝7.1Hz,J 2＝7.8Hz,1H,CH),0.89(s,3H,CH 3),0.87(s,3H,CH 3),0.83(s,3H,CH 3).ESI-MS:526.7[M+H] +.Anal.Calcd for C 30H 43N 3O 5.
Embodiment four:
MTT [3-(4,5)-bis-methyl-2-thiazole-(2,5)-phenyl bromination tetrazole is blue] method is adopted to measure pyrazoline hydroxamic acid C 21steroidal saponin aglycone derivative is to the half-inhibition concentration (IC of human breast cancer cell (MCF-7), cervical cancer cell (HeLa), lung carcinoma cell (A549) and liver cancer cell (HepG2) 50).
(1) preparation of nutrient solution (/L): 1. suspension cell: DMEM cultivates one bag, powder (10.4g), new-born calf serum 100mL, penicillin solution (2 × 10 -5u/mL) 0.5mL, Streptomycin Solution (2 × 10 -5u/mL) 0.5mL, after adding tri-distilled water dissolving, with the NaHCO of 5.6% 3solution adjust pH, to 7.2-7.4, is finally settled to 1000mL.Filtration sterilization.2. attached cell: the same, then add NaHCO 32.00g, HEPES 2.38g.
(2) preparation of D-Hanks damping fluid (often liter): NaCl 8.00g, KCl 0.40g, Na 2hPO 412H 2o 0.06g, KH 2pO 40.06g, NaHCO 30.35g.Autoclaving.
(3) preparation of trypsin solution: utilizing D-Hanks damping fluid to be made into concentration is 0.5% trypsin solution.Filtration sterilization.
(4) preparation of liquid is tested: dissolved by a small amount of tri-distilled water of test sample and be made into storing solution, i.e. 10 times of preparation storing solutions of empirically maximum concentration.Different according to compound dissolution, available tri-distilled water directly dissolves, or with a small amount of DMSO hydrotropy, then adds tri-distilled water and dissolve.Storing solution is stored in-20 DEG C of refrigerators for subsequent use.
(5) cultivation of human breast cancer cell (MCF-7), cervical cancer cell (HeLa), lung carcinoma cell (A549) and liver cancer cell (HepG2): be adherent growth cell, cellar culture is (containing 10% calf serum, 100U/mL Streptomycin sulphate) in DMEM nutrient solution, is placed in 37 DEG C, 5%CO 2cultivate in incubator, go down to posterity once every 3-4d.When going down to posterity, nutrient solution in former bottle is transferred in centrifuge tube, the centrifugal 5min of 1000rpm, discard original fluid, add equivalent fresh medium, piping and druming evenly, pipette appropriate in fresh culture bottle, then supplement fresh medium to original volume (nutrient solution volume is about 1/10 of culturing bottle capacity).
(6) cell incubation: the tumour cell in vegetative period of taking the logarithm, tune concentration of cell suspension is 1-1.5 × 10 5individual/mL.In 96 well culture plates, every hole adds cell suspension 100 μ L, puts 37 DEG C, 5%CO 224h is cultivated in incubator.After cultivating 24h, add liquid by design respectively.
(7) dosing: join in each hole by test liquid respectively according to the concentration gradient of ultimate density, each concentration establishes 6 parallel holes.Experiment is divided into drug test group (adding the test medicine of different concns respectively), control group (only add nutrient solution and cell, do not add test medicine) and blank group (only add nutrient solution, do not add cell and test medicine).96 orifice plates after dosing are placed in 37 DEG C, 5%CO 248h is cultivated in incubator.The activity of positive control medicine measures according to the method for test sample.
(8) mensuration of survivaling cell: in 96 orifice plates after having cultivated 48h, every hole adds MTT 40 μ L (being made into 4mg/mL with D-Hanks damping fluid).After placing 4h at 37 DEG C, remove supernatant liquor.Every hole adds 150 μ L DMSO, and vibration 5min, makes formazan dissolving crystallized.Finally, automatic microplate reader is utilized to detect the optical density(OD) (OD value) in each hole at 570nm wavelength place.
Half-inhibition concentration (IC 50) be defined as drug level when the tumor cell survival of 50%.According to the optical density(OD) (OD value) measured, make the typical curve of inhibitory rate of cell growth, typical curve is tried to achieve the drug level of its correspondence.
The IC recorded 50be shown in Table 1.
The listed novel pyrazoline hydroxamic acid C of table 1 the present invention 21steroidal saponin aglycone derivative is to the suppression IC of tumour cell 50value (μm ol/mL)
a6 parallel tests, experimental result is averaged, and error is between 5-10%.
Pyrazoline hydroxamic acid C of the present invention 21steroidal saponin aglycone derivative has obvious restraining effect to human breast cancer cell (MCF-7), cervical cancer cell (HeLa), lung carcinoma cell (A549) and liver cancer cell (HepG2), the inhibit activities of contrast Tenacissoside A is significantly improved, and contrast positive control drug then shows quite or is better than the inhibit activities of positive control medicine.Therefore pyrazoline hydroxamic acid C of the present invention 21steroidal saponin aglycone derivative can be applied to prepares antitumor drug.
Embodiment six:
The present invention tests new synthetic compound 18-21 to the cytotoxicity of people's renal epithelial cell (293T), cytotoxicity result as table 2, using Celecoxib as positive control.The toxicity suppressor T cell survival rate of each compound to 50% time concentration (CC 50) represent.
Experimental technique:
(1) cultivator renal epithelial cell (293T) is tending towards fusion until reach its logarithmic growth end of term cell, digests cell dispersion, be mixed with 1 with cell culture fluid with cell dissociation buffer
× 10 4the cell suspension of individual/mL.Get 96 well culture plates, in every hole, add the cell suspension of 100 μ L.Horizontally rotating culture plate gently makes cell be evenly dispersed in the surface in ware hole.
(2) be placed in containing 5%CO 2in cell culture incubator, at 37 ± 2 DEG C of temperature, cultivate 24h.Discard original fluid, every hole adds the blank liquid of 100 μ L, negative controls, positive control solution, the test sample vat liquor of 100% and 50% concentration.Often organize and at least establish 8 holes.Note: lixiviate stoste or make the serial lixiviate diluent of thinner with substratum.When adopting 0.9% sodium chloride injection lixiviate, use 2 times of concentrated substratum when diluting lixiviate.
(3) be placed in containing 5%CO 2in incubator, cultivate at 37 ± 2 DEG C of temperature.Cultivate 48h.
(4) after date between each cultivation, every hole adds MTT solution 20 μ L, is placed in containing 5%CO 2in incubator, at 37 ± 2 DEG C of temperature, cultivate 5h.
(5) discard liquid in hole, every hole adds 200 μ L DMSO respectively, and culture plate is placed 10min, and level is rocked and made solution colour in hole even.
(6) measure absorbancy by microplate reader, wavelength adopts 570nm.
The CC recorded 50be shown in Table 2.
The listed novel pyrazoline hydroxamic acid C of table 2 the present invention 21steroidal saponin aglycone derivative is to the suppression CC of 293T cell 50value (μm ol/mL)
a6 parallel tests, experimental result is averaged, and error is between 5-10%
From the above: pyrazoline hydroxamic acid C of the present invention 21steroidal saponin aglycone derivative is to showing quite people's renal epithelial cell (293T) or being better than the cytotoxicity of positive control medicine, and compare with Tenacissoside A, cytotoxicity also almost maintains an equal level.Therefore pyrazoline hydroxamic acid C of the present invention 21steroidal saponin aglycone derivative can be applied to prepares antitumor drug.
Contriver thinks: hydroxamic acid structural formula is R-CO-NHOH, is a kind of organic sequestering agent, energy and Cu 2+, Zn 2+, Co 2+, Ni 2+, Fe 3+ions etc. form stable metallo-chelate, and it has significant complex ability.Pyrazoline is very important nitrogenous five member ring heterocyclic compound, and it has excellent anti-tumor activity.
In a word, the present invention will have outstanding bioactive hydroxamic acid skeleton and be incorporated in pyrazoline derivative, combine simultaneously and have C active very well 21steroidal saponin aglycon skeleton, a series of pyrazoline hydroxamic acid C of design and synthesis 21steroidal saponin aglycone derivative, find through experiment, it has the advantages such as better biological activity, higher selectivity and lower toxicity.
More than describe the preferred embodiment of the present invention in detail; but the present invention is not limited to the detail in above-mentioned embodiment, within the scope of technical conceive of the present invention; can carry out multiple equivalents to technical scheme of the present invention, these equivalents all belong to protection scope of the present invention.It should be noted that in addition, each the concrete technical characteristic described in above-mentioned embodiment, in reconcilable situation, can be combined by any suitable mode.In order to avoid unnecessary repetition, the present invention illustrates no longer separately to various possible array mode.In addition, also can carry out arbitrary combination between various different embodiment of the present invention, as long as it is without prejudice to thought of the present invention, it should be considered as content disclosed in this invention equally.

Claims (10)

1. structure is such as formula the compound shown in VIII,
R 1for H, CH 3, CH 2cH 3.
2. structure is such as formula a preparation method for the compound shown in VIII,
R 1for H, CH 3, CH 2cH 3.
It is characterized in that, comprise the steps:
Step 1: prepare hydrazino-benzoic acid methyl esters, and by structure such as formula the converting compounds shown in I for structure is such as formula the compound shown in II;
Step 2: add diazomethane ether mixed solution, boron trifluoride-ether complex and structure such as formula the compound shown in II in reactor, be obtained by reacting the compound of structure as shown in formula III;
Step 3: add the chemical combination of structure as shown in formula III, the KOH aqueous solution, structure such as formula the compound shown in IV and ethanol in reactor, obtain structure after reaction such as formula the compound shown in V;
Step 4: by structure such as formula the compound dehydrated alcohol shown in V, join in reaction vessel to hydrazino-benzoic acid methyl esters and Glacial acetic acid, be obtained by reacting structure such as formula the compound shown in IV;
Step 5: join in the hydroxylamine solution of activation such as formula the compound shown in IV by structure, is obtained by reacting structure such as formula the compound shown in VII;
Step 6: add structure such as formula the compound shown in VII, methylene dichloride in reactor, and drip boron tribromide, be obtained by reacting target product;
Formula IV ~ VII, R 1for H, CH 3, CH 2cH 3.
3. the preparation method of compound as claimed in claim 1, it is characterized in that, described step 1 is further:
At 0 ± 5 DEG C, add in reactor successively anhydrous methanol, sulfur oxychloride, to hydrazino-benzoic acid, after stirring 30 ~ 40min, continue at normal temperatures to stir 3-5h, the solid filtering gained uses distilled water, ethanol, distilled water wash successively, dry, and the solid product obtained is dissolved in dehydrated alcohol, recrystallization is purified, and obtains hydrazino-benzoic acid methyl esters.
Under stirring at room temperature, structure is added successively such as formula the compound shown in I, aqueous hydrochloric acid, dehydrated alcohol in reactor, after stirring 2 ~ 3h, back flow reaction 7 ~ 9h, TLC follow the tracks of reaction, after reaction terminates, filter the solid obtained and use aqueous hydrochloric acid, distilled water, ethanol, distilled water wash successively, dry, the solid crude product obtained is dissolved in dehydrated alcohol, recrystallization is purified, and obtains structure such as formula the compound shown in II.
4. the preparation method of compound as claimed in claim 1, it is characterized in that, described step 2 is further:
Under ice-water bath stirring action, diazomethane ether mixed solution, boron trifluoride-ether complex is added successively in reactor, and the compound of structure as shown in formula III, after reaction 20 ~ 40min, back flow reaction 5-7h, TLC follows the tracks of reaction, and after reaction terminates, filtration obtains solid and uses aqueous hydrochloric acid, distilled water, ethanol, distilled water wash successively, dry, the solid product obtained is dissolved in dehydrated alcohol, and recrystallization is purified, and obtains the compound of structure as shown in formula III.
5. the preparation method of compound as claimed in claim 1, it is characterized in that, described step 3 is further:
Under stirring at room temperature, the compound of structure as shown in formula III, the KOH aqueous solution, structure is added successively such as formula the compound shown in IV, ethanol in reactor, after continuing stirring reaction 3 ~ 6h, pH is regulated with hcl acidifying, filter the solid obtained and use distilled water, ethanol, distilled water wash successively, the dry structure that obtains is such as formula the compound shown in V.
6. the preparation method of compound as claimed in claim 1, it is characterized in that, described step 4 is further:
Successively by structure such as formula the compound shown in V, dehydrated alcohol, join in reactor to hydrazino-benzoic acid methyl esters and Glacial acetic acid, after stirring at room temperature reaction 20 ~ 40min, have fraction solids insoluble; Back flow reaction 5 ~ 8h, TLC follow the tracks of reaction, after reaction terminates, filter, solid uses aqueous hydrochloric acid, distilled water, ethanol, distilled water wash successively, dry, the solid product obtained is dissolved in dehydrated alcohol, and recrystallization is purified, and obtains structure such as formula the compound shown in VI.
7. the preparation method of compound as claimed in claim 1, it is characterized in that, described step 5 is further:
Hydrochloric acid hydroxyl is added in anhydrous methanol by, sodium methylate, after heated and stirred 20 ~ 40min, filters solid, obtain the hydroxylamine solution activated, add structure wherein such as formula the compound shown in VI, back flow reaction 3 ~ 5h, TLC follows the tracks of reaction, after reaction terminates, filter, the solid distilled water wash obtained, last vacuum-drying, the solid obtained is dissolved in dehydrated alcohol, and recrystallization is purified, and obtains structure such as formula the compound shown in VII.
8. the preparation method of compound as claimed in claim 1, it is characterized in that, described step 6 is further:
Under stirring at-20 ± 5 DEG C, in reactor, add structure successively such as formula the compound shown in VII, methylene dichloride, and progressively drip boron tribromide, stir 20 ~ 40min, room temperature continues reaction 8 ~ 12h, and TLC follows the tracks of reaction, after reaction terminates, filter, solid distilled water wash, last vacuum-drying, is dissolved in dehydrated alcohol by the solid obtained, recrystallization is purified, and obtains lenticular target compound.
9. structure is preparing the application in cancer therapy drug such as formula the compound shown in VIII,
Wherein, R 1for H, CH 3, CH 2cH 3.
10. an anticancer pharmaceutical composition, is characterized in that, comprises structure such as formula the compound shown in VIII and medically acceptable carrier,
Wherein, R 1for H, CH 3, CH 2cH 3.
CN201510219524.4A 2015-04-30 2015-04-30 Novel anti-cancer compound, preparation method for anti-cancer compound and application of anti-cancer compound to preparation of anti-cancer drugs Pending CN104804066A (en)

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