CN104558093A - C21 steroid saponin aglycone derivative as well as preparation method and application thereof in preparing anti-tumor drugs - Google Patents

C21 steroid saponin aglycone derivative as well as preparation method and application thereof in preparing anti-tumor drugs Download PDF

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CN104558093A
CN104558093A CN201510059100.6A CN201510059100A CN104558093A CN 104558093 A CN104558093 A CN 104558093A CN 201510059100 A CN201510059100 A CN 201510059100A CN 104558093 A CN104558093 A CN 104558093A
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formula
reaction
compound shown
distilled water
solid
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CN104558093B (en
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严晓强
朱海亮
杨永安
钟飞
俞海荣
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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 C21 steroid saponin aglycone derivative as well as a preparation method and an application thereof in preparing anti-tumor drugs. The structural formula of the C21 steroid saponin aglycone derivative is as shown in formula IX. The C21 steroid saponin aglycone derivative disclosed by the invention has the advantages of being better in bioactivity, higher in selectivity, lower in toxicity and the like. The C21 steroid saponin aglycone derivative has a significant inhibitory effect on human breast cancer cells, cervical cancer cells, lung cancer cells and liver cancer cells (as shown in Specification), wherein R1 is selected from H and F, R2 is selected from H and F, and R3 is selected from H and F.

Description

C 21steroidal saponin aglycone derivative, its preparation method and preparing the application in antitumor drug
Technical field
The invention belongs to medicinal chemistry art, especially a kind of C 21steroidal saponin aglycone derivative, its preparation method and preparing the application in antitumor drug.
Background technology
From Chinese medicine Caulis Marsdeniae Tenacissimae, can obtain steroidal saponin constituents through extraction separation and purification, 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 C 21steroidal saponin aglycone derivative, structure such as formula shown in IX,
Wherein, R 1be selected from H, F, R 2be selected from H, F, R 3be selected from H, F.
A kind of C 21the preparation method of steroidal saponin aglycone derivative,
Described C 21the structure of steroidal saponin aglycone derivative such as formula shown in IX,
Wherein, R 1be selected from H, F, R 2be selected from H, F, R 3be selected from H, F;
Comprise the steps:
Step 1, prepare hydrazino-benzoic acid methyl esters, add structure such as formula the compound shown in I, aqueous hydrochloric acid and dehydrated alcohol in reactor, after reaction, obtained structure is such as formula the compound shown in II;
Step 2, in reactor, add diazomethane ether mixed solution, boron trifluoride-ether complex and structure such as formula the compound shown in II, after reaction, obtain the compound of structure as shown in formula III;
Step 3, in reactor, add the compound of structure as shown in formula III, the KOH aqueous solution, structure such as formula the compound shown in IV and ethanol, obtained structure is such as formula the compound shown in V;
Step 4, 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, be obtained by reacting structure such as formula the compound shown in VI;
Step 5, structure is joined in the mixing solutions of methyl alcohol, THF such as formula the compound shown in VI, KOH, obtain structure after reaction such as formula the compound shown in VII;
Step 6, structure is joined in methylene dichloride such as formula the compound shown in VII and N methyl piperazine, mixing is dissolved, and drips the methylene dichloride mixing solutions containing DCC/DMAP, is obtained by reacting structure such as formula the compound shown in VIII.
Step 7, in reactor, add structure such as formula the compound shown in VIII, methylene dichloride, and drip boron tribromide, produce target compound;
In formula IV ~ VIII, R 1be selected from H, F, R 2be selected from H, F, R 3be selected from H, F.
In a preferred embodiment, it is characterized in that, described step 1 is further:
At 0 ± 5 DEG C, anhydrous methanol, sulfur oxychloride and to hydrazino-benzoic acid is added successively in reactor, after stirring 30 ~ 40min, normal temperature continues to stir 3-5h, and filter, the solid obtained uses distilled water, ethanol and distilled water wash successively, dry, the solid product obtained is dissolved in dehydrated alcohol, and recrystallization is purified, and obtains carboxylate to 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 follows the tracks of reaction, after reaction terminates, filter, the solid obtained uses aqueous hydrochloric acid, distilled water, cold ethanol, distilled water wash successively, dry, the solid crude product obtained is dissolved in dehydrated alcohol, and recrystallization is purified, and obtains the structure of lenticular such as formula the compound shown in II.
Preferred, I 10mmol is dissolved in 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, structure is added successively such as formula the compound shown in II in reactor, after reaction 20 ~ 40min, then back flow reaction 5 ~ 7h, TLC follows the tracks of reaction, after reaction terminates, filter, obtain solid and use 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 the compound of structure as shown in formula III.
Preferred, each reactant molar ratio: II: diazomethane: boron tribromide=1:1.2 ~ 1.5:1.5 ~ 2.0.Preferred further, ratio is 1:1.2:2.0,1:1.4:1.5.
In a preferred embodiment, described step 3 is further:
In room temperature (herein, room temperature refers to 15 DEG C ~ 25 DEG C) stir under, 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 and ethanol in reactor, after continuing stirring reaction 3 ~ 6h, with hcl acidifying, regulate pH, filter, the solid obtained uses distilled water, cold ethanol, distilled water wash successively, obtains structure such as formula the compound shown in V after drying.
Preferred, each reactant molar ratio: IV:III: boron tribromide=1:1:1 ~ 1.5.Preferred further, ratio is 1:1:1.5,1:1:1.
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, still 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, cold 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, mol ratio: 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 further for: structure is joined such as formula the compound shown in VI and KOH in the mixing solutions of methyl alcohol, THF water, back flow reaction 4 ~ 8h, TLC follows the tracks of reaction, after reaction terminates, with hcl acidifying, regulate pH to acid, solution is poured in saturated aqueous common salt, filter, solid distilled water wash, 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, volume ratio: methyl alcohol: THF: water=1:1:1.The structure of 10mmol is dissolved in mol ratio: VI:KOH=1:2 in 50ml methyl alcohol THF water mixed liquid such as formula the compound shown in VI.
In a preferred embodiment, described step 6 is further:
Structure is joined in methylene dichloride such as formula the compound shown in VII and N methyl piperazine, mixing is dissolved, the methylene dichloride mixing solutions containing DCC/DMAP is dripped under stirring at 0 ± 5 DEG C DEG C, reaction 20 ~ 40min, then 6 ~ 10h is reacted under stirring at 35 ± 5 DEG C, TLC follows the tracks of reaction, after reaction terminates, reaction solution is poured in saturated aqueous common salt, filter, solid uses aqueous hydrochloric acid, distilled water, cold ethanol, distilled water wash successively, dry, the solid product obtained is dissolved in dehydrated alcohol, and recrystallization is purified and is obtained structure such as formula the compound shown in VIII.
Preferred, in above-mentioned steps, the mol ratio of each reactant: VII:N-methylpiperazine: DCC:DMAP=1:1:1.2:1.44.
In a preferred embodiment, described step 7 is further:
Under stirring at-20 ± 5 DEG C, in reactor, add structure successively such as formula the compound shown in VIII, methylene dichloride, and 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 the target compound of lenticular.
Preferred, in above-mentioned steps, the mol ratio of each reactant: VIII: boron tribromide=1:0.5.
Present method is transformed for saponin(e aglycon, has effectively integrated piperazine 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.
A kind of C 21steroidal saponin aglycone derivative is preparing the application in antitumor drug,
Described C 21the structure of steroidal saponin aglycone derivative such as formula shown in IX,
Wherein, R 1be selected from H, F, R 2be selected from H, F, R 3be selected from H, F.
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
Technical scheme of the present invention is as follows:
The pyrazoline piperazine C of one class novelty 21the synthesis of steroidal saponin aglycone derivative, is characterized in that it has following general formula:
Pyrazoline piperazine C of the present invention 21the synthesis of steroidal saponin aglycone derivative, comprises the following steps:
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 (a kind of C successively in round-bottomed flask 21steroidal saponin) 3, aqueous 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 6-9.
Step 5., successively by different compound 6-9, 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 follows 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 10-13 by the solid crude product obtained.
Compound 10-13, KOH join in the mixing solutions of methyl alcohol, THF water by step 6., after about back flow reaction 5h.TLC follows the tracks of reaction (developping agent V acOEt: V pE=1:2), after reaction terminates, dilute hydrochloric acid acidifying regulates pH to acid, is poured into by solution in saturated aqueous common salt, filter, solid distilled water wash, last vacuum-drying, the solid obtained is dissolved in dehydrated alcohol, and recrystallization is purified, and obtains lenticular compound 14-17.
Step 7. joins compound 14-17 and N methyl piperazine in methylene dichloride, and mixing is dissolved, and drips the methylene dichloride mixing solutions containing DCC/DMAP at about 0 DEG C under stirring, and after reaction about 0.5h, then reacts about 8h under stirring at 35 ± 5 DEG C.TLC follows the tracks of reaction (developping agent V acOEt: V pE=1:2), after reaction terminates, reaction solution is poured in saturated aqueous common salt, 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 18-21 by the solid crude product obtained.
Step 8. adds corresponding intermediate 18-21, 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 22-25.
Embodiment one:
N-methyl-4-(5-phenyl-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 piperazine (compound 22)
Under stirring at-20 DEG C, corresponding intermediate 18 (10.0mmol), methylene dichloride (25mL) that step 7 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 22.
Obtain white crystal, productive rate 47.3%.m.p.221～223℃； 1H NMR(DMSO-d 6,300MHz)δ:7.98(d,J＝8.1Hz,2H,ArH),7.68(d,J＝7.7Hz,2H,ArH),7.36～7.25(m,5H,ArH),5.37(s,2H,OH),5.19(t,J＝7.3Hz,1H,CH),4.49(s,1H,OH),3.58～3.42(m,6H,CH and CH 2),3.34(d,J＝6.8Hz,1H,CH),,2.95～2.73(m,2H,CH 2),2.31(t,J＝7.7Hz,4H,CH 2),2.18(s,3H,CH 3),2.01(t,J＝5.8Hz,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:669.9[M+H] +.Anal.Calcd forC 40H 52N 4O 5:C,H,N.
Embodiment two:
N-methyl-4-(5-(the fluoro-phenyl of 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 piperazine (compound 23)
Preparation method's reference example one.Obtain white crystal, productive rate 45.5%.m.p.226～227℃； 1H NMR(DMSO-d 6,300MHz)δ:8.03(d,J＝8.3Hz,2H,ArH),7.73(d,J＝8.1Hz,2H,ArH),7.35～7.18(m,4H,ArH),5.35(s,2H,OH),5.19(t,J＝7.1Hz,1H,CH),4.49(s,1H,OH),3.56～3.41(m,6H,CH and CH 2),3.32(d,J＝6.8Hz,1H,CH),2.97～2.71(m,2H,CH 2),2.28(t,J＝7.7Hz,4H,CH 2),2.15(s,3H,CH 3),2.04(t,J＝5.7Hz,1H,CH),1.81～1.20(m,15H,CHand CH 2),1.14(dd,J 1＝7.0Hz,J 2＝7.6Hz,1H,CH),0.89(s,3H,CH 3),0.78(s,3H,CH 3).ESI-MS:687.9[M+H] +.Anal.Calcd for C 40H 51FN 4O 5:C,H,N.
Embodiment three:
N-methyl-4-(5-(the fluoro-phenyl of 2-)-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 piperazine (compound 24).
Preparation method's reference example one.Obtain white crystal, productive rate 44.9%.m.p.212～213℃； 1H NMR(DMSO-d 6,300MHz)δ:10.56(s,1H,OH),8.03(d,J＝8.1Hz,2H,ArH),7.74～7.55(m,5H,ArH and NH),7.29～7.23(m,1H,ArH),7.15～7.07(m,1H,ArH),5.37(s,2H,OH),5.19(t,J＝7.3Hz,1H,CH),4.48(s,1H,OH),3.55～3.42(m,6H,CH and CH 2),3.34(d,J＝6.8Hz,1H,CH),2.95～2.73(m,2H,CH 2),2.32(t,J＝7.7Hz,4H,CH 2),2.01(t,J＝5.4Hz,1H,CH),1.79～1.21(m,15H,CH and CH 2),1.14(dd,J 1＝7.1Hz,J 2＝7.9Hz,1H,CH),0.84(s,3H,CH 3),0.81(s,3H,CH 3).ESI-MS:687.9[M+H] +.Anal.Calcd for C 40H 51FN 4O 5:C,H,N.
Embodiment four:
N-methyl-4-(5-(the fluoro-phenyl of 3-)-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 piperazine (compound 25).
Preparation method's reference example one.Obtain white crystal, productive rate 37.8%.m.p.215～216℃； 1H NMR(DMSO-d 6,300MHz)δ:10.49(s,1H,OH),7.99(d,J＝8.1Hz,2H,ArH),7.72(d,J＝7.3Hz,2H,ArH),7.66(s,1H,NH),7.39～7.31(m,1H,ArH),7.15～7.07(m,2H,ArH),6.97(d,J＝3.9Hz,1H,ArH),5.35(s,2H,OH),5.16(t,J＝7.2Hz,1H,CH),4.49(s,1H,OH),3.54～3.41(m,6H,CH and CH 2),3.34(d,J＝6.5Hz,1H,CH),,2.95～2.73(m,2H,CH 2),2.30(t,J＝7.7Hz,4H,CH 2),2.01(t,J＝5.5Hz,1H,CH),1.75～1.20(m,15H,CH and CH 2),1.13(dd,J 1＝6.6Hz,J 2＝7.2Hz,1H,CH),0.85(s,3H,CH 3),0.83(s,3H,CH 3).ESI-MS:687.9[M+H] +.Anal.Calcd for C 40H 51FN 4O 5:C,H,N.
Embodiment five:
MTT [3-(4,5)-bis-methyl-2-thiazole-(2,5)-phenyl bromination tetrazole is blue] method is adopted to measure pyrazoline piperazine 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 piperazine 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%
From the above: pyrazoline piperazine 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), especially best to A549 cell, 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 piperazine 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 and Tenacissoside A 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 the cell suspension of 1 × 104/mL with cell culture fluid with cell dissociation buffer.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%CO2 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%CO2 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%CO2 incubator, at 37 ± 2 DEG C of temperature, cultivates 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 piperazine 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%.
Pyrazoline piperazine 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 piperazine C of the present invention 21steroidal saponin aglycone derivative can be applied to prepares antitumor drug.
Contriver thinks: hexahydropyrazine is the hexa-member heterocycle containing 2 nitrogen-atoms in molecule, and its Enthalpies of Formation is high, Heat stability is good, is the ideal building blocks of azotic heterocyclic compound, has good nitrogen equilibrium symmetrical structure.Introduce piperazine ring in the present invention, as the synergist groups of medicine, the pharmacokinetic property of medicine can be improved, improve the biological activity of medicine.And it also has that onset of action is fast, side reaction is little, toxicity is low, without the feature such as additive.Pyrazoles is the important heterogeneous ring compound of a class, has efficient, low toxicity, and the character of the multi-faceted conversion of its ring substituents.
In a word, piperazine skeleton is incorporated in pyrazoline derivative by the present invention, combines simultaneously and has C active very well 21steroidal saponin aglycon skeleton, a series of pyrazoline piperazine C of design and synthesis 21steroidal saponin aglycone derivative, experimentally result, 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. a C 21steroidal saponin aglycone derivative, is characterized in that, structure such as formula shown in IX,
Wherein, R 1be selected from H, F, R 2be selected from H, F, R 3be selected from H, F.
2. a C 21the preparation method of steroidal saponin aglycone derivative,
Described C 21the structure of steroidal saponin aglycone derivative such as formula shown in IX,
Wherein, R 1be selected from H, F, R 2be selected from H, F, R 3be selected from H, F;
It is characterized in that, comprise the steps:
Step 1, prepare hydrazino-benzoic acid methyl esters, add structure such as formula the compound shown in I, aqueous hydrochloric acid and dehydrated alcohol in reactor, after reaction, obtained structure is such as formula the compound shown in II;
Step 2, in reactor, add diazomethane ether mixed solution, boron trifluoride-ether complex and structure such as formula the compound shown in II, after reaction, obtain the compound of structure as shown in formula III;
Step 3, in reactor, add the compound of structure as shown in formula III, the KOH aqueous solution, structure such as formula the compound shown in IV and ethanol, obtained structure is such as formula the compound shown in V;
Step 4, 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, be obtained by reacting structure such as formula the compound shown in VI;
Step 5, structure is joined in the mixing solutions of methyl alcohol, THF such as formula the compound shown in VI, KOH, obtain structure after reaction such as formula the compound shown in VII;
Step 6, structure is joined in methylene dichloride such as formula the compound shown in VII and N methyl piperazine, mixing is dissolved, and drips the methylene dichloride mixing solutions containing DCC/DMAP, is obtained by reacting structure such as formula the compound shown in VIII.
Step 7, in reactor, add structure such as formula the compound shown in VIII, methylene dichloride, and drip boron tribromide, produce target compound;
2 -->
In formula IV ~ VIII, R 1be selected from H, F, R 2be selected from H, F, R 3be selected from H, F.
3. C as claimed in claim 2 21the preparation method of steroidal saponin aglycone derivative, is characterized in that, described step 1 is further:
At 0 ± 5 DEG C, anhydrous methanol, sulfur oxychloride and to hydrazino-benzoic acid is added successively in reactor, after stirring 30 ~ 40min, continue at normal temperatures to stir 3-5h, filter, the solid obtained uses distilled water, ethanol and distilled water wash successively, dry, the solid product obtained is dissolved in dehydrated alcohol, and recrystallization is purified, and obtains carboxylate to 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 follows the tracks of reaction, after reaction terminates, filter, the solid obtained uses aqueous hydrochloric acid, distilled water, cold ethanol, distilled water wash successively, dry, the solid crude product obtained is dissolved in dehydrated alcohol, and recrystallization is purified, and obtains the structure of lenticular such as formula the compound shown in II.
4. C as claimed in claim 2 21the preparation method of steroidal saponin aglycone derivative, is characterized in that, described step 2 is further:
Under ice-water bath stirring action, diazomethane ether mixed solution, boron trifluoride-ether complex, structure is added successively such as formula the compound shown in II in reactor, after reaction 20 ~ 40min, back flow reaction 5-7h, TLC follows the tracks of reaction, after reaction terminates, filter, obtain solid and use 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 the compound of structure as shown in formula III.
5. C as claimed in claim 2 21the preparation method of steroidal saponin aglycone derivative, 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 and ethanol in reactor, after continuing stirring reaction 3 ~ 6h, with hcl acidifying, regulate pH, filter, the solid obtained uses distilled water, cold ethanol, distilled water wash successively, obtains structure such as formula the compound shown in V after drying.
6. C as claimed in claim 2 21the preparation method of steroidal saponin aglycone derivative, 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, cold 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. C as claimed in claim 2 21the preparation method of steroidal saponin aglycone derivative, it is characterized in that, described step 5 further for: structure is joined such as formula the compound shown in VI and KOH in the mixing solutions of methyl alcohol, THF water, back flow reaction 4 ~ 8h, TLC follows the tracks of reaction, after reaction terminates, with hcl acidifying, regulate pH to acid, solution is poured in saturated aqueous common salt, filter, solid distilled water wash, last vacuum-drying, is dissolved in dehydrated alcohol by the solid obtained, recrystallization is purified, and obtains structure such as formula the compound shown in VII.
8. C as claimed in claim 2 21the preparation method of steroidal saponin aglycone derivative, is characterized in that, described step 6 is further:
Structure is joined in methylene dichloride such as formula the compound shown in VII and N methyl piperazine, mixing is dissolved, the methylene dichloride mixing solutions containing DCC/DMAP is dripped under stirring at 0 ± 5 DEG C, reaction 20 ~ 40min, then 6 ~ 10h is reacted under stirring at 35 ± 5 DEG C, TLC follows the tracks of reaction, after reaction terminates, reaction solution is poured in saturated aqueous common salt, filter, solid uses aqueous hydrochloric acid, distilled water, cold ethanol, distilled water wash successively, dry, the solid product obtained is dissolved in dehydrated alcohol, and recrystallization is purified and is obtained structure such as formula the compound shown in VIII.
9. C as claimed in claim 2 21the preparation method of steroidal saponin aglycone derivative, is characterized in that, described step 7 is further:
Under stirring at-20 ± 5 DEG C, in reactor, add structure successively such as formula the compound shown in VIII, methylene dichloride, and 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 the target compound of lenticular.
10. a C 21steroidal saponin aglycone derivative is preparing the application in antitumor drug, described C 21the structure of steroidal saponin aglycone derivative such as formula shown in IX,
Wherein, R 1be selected from H, F, R 2be selected from H, F, R 3be selected from H, F.
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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104804062A (en) * 2015-04-30 2015-07-29 江苏耐雀生物工程技术有限公司 Pyrazoline piperazine steroid saponin aglycone derivative containing naphthalene skeleton, and preparation method and application of derivative
CN104829683A (en) * 2015-04-30 2015-08-12 江苏耐雀生物工程技术有限公司 Indole skeleton-containing dihydropyrazolpiperazine C21 steroid sapogenin derivative, and preparation method and application thereof
CN104829682A (en) * 2015-04-29 2015-08-12 江苏耐雀生物工程技术有限公司 Furan-skeleton-containing 2H-pyrazole piperazine C21 steroidal saponin aglycone derivatives, preparation method, and applications thereof
CN104829534A (en) * 2015-05-20 2015-08-12 南京大学 Preparation method of dihydro-pyrazole morpholine derivatives containing naphthalene nucleus frameworks and application of dihydro-pyrazole morpholine derivatives to preparation of antitumor drugs
CN104861026A (en) * 2015-04-30 2015-08-26 江苏耐雀生物工程技术有限公司 Furan skeleton included 2H-pyrazole hydroxamic acid C21 steroid saponin aglycone derivative and preparation method and application thereof
CN104877002A (en) * 2015-04-29 2015-09-02 江苏耐雀生物工程技术有限公司 C21 steroid sapogenin derivative of dihydro parazole piperazidine, and preparation method and application thereof
CN104961683A (en) * 2015-07-10 2015-10-07 南京大学 Preparation method of dihydropyrazol piperazine derivatives containing naphthalene ring skeleton and application in anti-cancer drugs
CN105037265A (en) * 2015-05-20 2015-11-11 南京大学 Preparation method of quinolinone derivative containing chalcone framework, and application of the quinolinone derivative in anti-cancer medicines

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101475623A (en) * 2009-02-18 2009-07-08 江苏先声药物研究有限公司 Steroid compounds with endothelin receptor A antagonistic activity and intermediate thereof
WO2009090063A1 (en) * 2008-01-16 2009-07-23 Jado Technologies Gmbh Steroid sapogenin, androstane and triterpenoid sapogenin derivatives for the treatment and prevention of infectious diseases

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009090063A1 (en) * 2008-01-16 2009-07-23 Jado Technologies Gmbh Steroid sapogenin, androstane and triterpenoid sapogenin derivatives for the treatment and prevention of infectious diseases
CN101475623A (en) * 2009-02-18 2009-07-08 江苏先声药物研究有限公司 Steroid compounds with endothelin receptor A antagonistic activity and intermediate thereof

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
ANABEL ROMERO-LOPEZ等: "Synthesis of steroidal derivatives containing substituted, fused and spiro pyrazolines", 《STEROIDS》 *
SHRAVANKUMAR KANKALA等: "Synthesis and anti-cancer evaluation of steroidal diglycoside-pyrazoline hybrids", 《RSC ADV.》 *
朱瑞静: "通光散肿瘤化疗增效有效部位的化学成分与质量控制方法研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 *

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104829682A (en) * 2015-04-29 2015-08-12 江苏耐雀生物工程技术有限公司 Furan-skeleton-containing 2H-pyrazole piperazine C21 steroidal saponin aglycone derivatives, preparation method, and applications thereof
CN104877002A (en) * 2015-04-29 2015-09-02 江苏耐雀生物工程技术有限公司 C21 steroid sapogenin derivative of dihydro parazole piperazidine, and preparation method and application thereof
CN104804062A (en) * 2015-04-30 2015-07-29 江苏耐雀生物工程技术有限公司 Pyrazoline piperazine steroid saponin aglycone derivative containing naphthalene skeleton, and preparation method and application of derivative
CN104829683A (en) * 2015-04-30 2015-08-12 江苏耐雀生物工程技术有限公司 Indole skeleton-containing dihydropyrazolpiperazine C21 steroid sapogenin derivative, and preparation method and application thereof
CN104861026A (en) * 2015-04-30 2015-08-26 江苏耐雀生物工程技术有限公司 Furan skeleton included 2H-pyrazole hydroxamic acid C21 steroid saponin aglycone derivative and preparation method and application thereof
CN104829534A (en) * 2015-05-20 2015-08-12 南京大学 Preparation method of dihydro-pyrazole morpholine derivatives containing naphthalene nucleus frameworks and application of dihydro-pyrazole morpholine derivatives to preparation of antitumor drugs
CN105037265A (en) * 2015-05-20 2015-11-11 南京大学 Preparation method of quinolinone derivative containing chalcone framework, and application of the quinolinone derivative in anti-cancer medicines
CN104961683A (en) * 2015-07-10 2015-10-07 南京大学 Preparation method of dihydropyrazol piperazine derivatives containing naphthalene ring skeleton and application in anti-cancer drugs

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