The purpose of this invention is to provide a series of preparation methods with the low compound of antitumour activity and toxicity, compound and as the application of medicine.
Dammarane-type saponin ginseng lowhydric alcohol derivative of the present invention has the general formula of formula I, and the formula I is:
In the formula I, R=H, COCH
2CH
2COOK, COCH
2CH
2COONa,
(abbreviation II), R '=II, H, OCOCH
2CH
2COONa.When R=H, R '=II, the represented compound of formula I is the panoxatriol glycoside, molecular formula C
36H
62O
9, white crystal.Work as R=COCH
2CH
2During COOK, R '=H, the represented compound of formula I is the panoxadiol potassium succinate, molecular formula C
34H
55O
6K, white crystal, variable color more than 360 ℃.Work as R=COCH
2CH
2During COONa, R '=OCOCH
2COONa, the represented compound of formula I is the panoxatriol sodium succinate, molecular formula C
38H
58O
10Na
2, white crystal, variable color more than 300 ℃.When the R=II, R '=H, the represented compound of formula I is the panoxadiol glycoside, molecular formula C
36H
62O
8, white crystal, fusing point 223-226 ℃.
Panoxatriol glycoside C
36H
62O
9The preparation method be: get panoxatriol 4-5g, dry ether 90-190ml, ethylene dichloride 85-120ml mixes, heating for dissolving adds acetyl bromide for glucose 6-12g, refluxes and stirs 4-10 hour, adding silver carbonate 4-7.9g backflow again stirred 4-10 hour, filter, filtrate is reclaimed solvent, and resistates is dissolved in methyl alcohol, filter, filtrate adds distilled water to the solution muddiness, adds the 6-12ml triethylamine again, stirring at room 8-14 hour, reclaim solvent to doing, by silica gel column chromatography,, get product with chloroform-methanol flushing in 9: 1.
Panoxadiol potassium succinate C
34H
55O
6The preparation method of K: a: with panoxadiol, succinyl oxide, pyridine is pressed 1-3: 5-6: the mixed of 15-20, heated and stirred refluxes, and room temperature is put cold, in the impouring cold water, have the precipitation separate out, filter the pale precipitation thing, throw out is dissolved in the ethanol, heating for dissolving adds activated carbon decolorizing, filtration under diminished pressure, reclaim ethanol, put coldly, an adularescent bunch shape crystallization is separated out, filter is done, and gets the panoxadiol succsinic acid; B, panoxadiol succsinic acid add stirring and dissolving in the acetone, drip the KOH ethanol solution, stir until precipitation fully, leave standstill, filter, and the throw out washing with acetone, filtration under diminished pressure gets product.
Panoxatriol sodium succinate C
38H
58O
10Na
2The preparation method be: a, panoxatriol, succinyl oxide, pyridine are pressed 1-2.5: the mixed of 3-6: 9-12, in 134-135 ℃ oil bath backflow 3-5 hour, then with in the reaction mixture impouring water, leave standstill, filter, drying, get pale solid, this solid is dissolved in the ethanol, with activated carbon decolorizing, filter, in the filtrate impouring water, leave standstill the leaching precipitate, make panoxatriol 3, the 6-disuccinic acid.
B, with panoxatriol-3, the 6-disuccinic acid is dissolved in the acetone, stir splash into the NaOH methanol solution to precipitation fully, filter, use washing with acetone, drying gets finished product.
Panoxatriol grape glucoside sugar of the present invention, panoxadiol potassium succinate, panoxatriol sodium succinate and panoxadiol glycoside (be called for short B ' 3) be through experiment, has the stronger ability that presses down the knurl effect and kill and wound cancer cells.The human cancer cell vitro culture fragmentation test data of above-claimed cpd see Table 1, the result shows that above-claimed cpd has the effect that kills and wounds lung adenocarcinoma cell, and significant concn-effect dependence is arranged, ultimate density is 100 μ g/ml when above, and kill rate can surpass 50%.3 pairs of Lewis lung cancer tumor-inhibiting actions of B ' are as shown in table 2, and the result shows that successive administration 7 days has tangible tumor-inhibiting action to Lewis liver cancer, and 50mg/kg group tumour inhibiting rate is 53.3%, reaches the rules requirement.Under equal conditions compare its tumor-inhibiting action and be lower than cis-platinum slightly with cis-platinum.3 couples of H of B '
22Liver ascites lengthen the life the effect as shown in table 3, the result shows, 7 days H of successive administration
22Liver cancer has the effect of obviously lengthening the life, and the 50mg/kg group can lengthen the life 155.3%, reaches the rules requirement, under equal conditions compares with cis-platinum, and its antitumor action and chemotherapeutic are suitable.3 pairs of mouse Emhorns of B ' EAS tumor-inhibiting action is as shown in table 4, and the result shows that successive administration 7 days has tangible tumor-inhibiting action to Emhorn EAS, and its action intensity and cis-platinum are suitable.
Dammarane-type saponin ginseng lowhydric alcohol derivative of the present invention can be made pharmaceutically acceptable dosage formulation respectively with 12-25mg/ml.
Embodiment:
1, the preparation of panoxatriol glycoside: panoxatriol 4.2 grams, acetyl bromide are for glucose 7.2 grams, 190 milliliters of ether (drying), 85 milliliters of ethylene dichloride, silver carbonate 7.9 grams
The preparation method:
Panoxatriol, dry ether, the dissolving of ethylene dichloride mixture heating up, add acetyl bromide for glucose, reflux and stirred 10 hours, add the silver carbonate backflow and stirred 10 hours, filter, reclaim solvent, resistates 20ml dissolve with methanol, the filtering insolubles, filtrate adds water to muddiness (adding the about 10-15ml of entry), adding the 12ml triethylamine stirred 12 hours, reclaim solvent to doing, be adsorbed on the 5 gram 160-200 order silica gel by 85 gram silica gel wet method upper props, with chloroform flushing in 9: 1,50 milliliters every part, 10-19 part is collected the panoxatriol glycoside.Weigh 0.94 gram.
2, the preparation of panoxatriol sodium succinate
(A) panoxatriol-3, the preparation of 6-disuccinic acid:
4.76 gram panoxatriol and 20 gram succinyl oxides are dissolved in the 50ml pyridine, reflux 3.5 hours in 134-135 ℃ oil bath, then with standing over night in the reaction mixture impouring 800ml water, filter, drying obtains 6.57 gram pale solids.This is dissolved among the 200ml95% ethanol,, filters with activated carbon decolorizing, in the filtrate impouring 800ml water (alcohol 1: water 4-5), standing over night is filtered, vacuum-drying in the vitriol oil vacuum drier, white solid 5.46 grams, productive rate 80.8%.
(B) preparation of panoxatriol sodium succinate:
3.0 the panoxatriol-3.6-disuccinic acid that restrains is dissolved in the 600ml propyl alcohol, under constantly stirring, splashes into the NaOH methanol solution of 10ml1N, drips to precipitating fully, continues to stir 1 hour, filters, vacuum-drying gets the 3.13g white solid.
3, the preparation of panoxadiol potassium succinate:
Panoxadiol 3 gram, succinyl oxide 6 grams, 15 milliliters of pyridines, in the 100ml triangular pyramidal bottle of packing into, stirring and refluxing in the water-bath react 2 hours, put coldly, in 300 milliliters of cold water of impouring, had precipitation to separate out, and decompress filter must white depositions.TLC reaction is a bit, and above-mentioned reactant is dissolved in 200ml(95%) in the ethanol, heating for dissolving, gac suitably decolours, and decompress filter reclaims ethanol to about 30ml, puts a cold adularescent bunch shape needle and separates out, and suction filtration, drying must the 3g products, productive rate 100%.
TLC condition: developping agent CHC13-MeOH(9: 1) developer 15% ethanol solution of sulfuric acid, 105 ℃ of heating.Thin layer plate silica gel G plate.
8 gram panoxadiol succsinic acids place the 500ml Erlenmeyer flask, add 400ml acetone, fully stir, slowly splash into 0.5mlKOH ethanol liquid, 30ml continues to stir 20 minutes altogether, leave standstill a moment, treat precipitation fully, filtration under diminished pressure, throw out washing with acetone 3 times, each 10ml, precipitation seasoning, porphyrize, weigh white powder 7.5gm, yield 94%.
4, the preparation of panoxadiol glycoside (B ' 3)
(A) preparation of Bromotetraacetylgluc,se:
In the three-necked bottle of 500ml, load onto stirring, add diacetyl oxide and a little vitriol oil, under the stirring state, gradation adds glucose, controlled temperature 60-80 ℃, add the back and continue to stir 30 fens, frozen water is cooled to below 30 ℃, and gradation adds red phosphorus, add the back and continue to stir 30 minutes, slowly drip Br below 30 ℃
2, after adding, the bottle stopper plug is good, puts refrigerator overnight.Add gauge water, with chloroform extraction, the chloroform layer washing adds CaC
12Drying is filtered, and filtrate decompression is regained chloroform, recrystallization in the resistates dehydrated alcohol, vacuum-drying.
(B) acetylize B ' 3 preparations:
Panoxadiol, acetylize bromo glucose, anhydrous diethyl ether; dichloroethane solution is heated to 40 ℃ in the water-bath, add silver carbonate; lucifuge stirred 8 hours; filter, reclaim the solvent residues thing and add 20ml methyl alcohol, remove by filter insolubles; evaporate to dryness methyl alcohol; last silicagel column with 8: 2 wash-outs of sherwood oil-acetone, gets acetylize B ' 3.Yield about 25%.
(3) preparation of B ' 3
Acetylize B ' 3 adds dissolve with methanol, drips to add water to little muddyly, adds triethylamine; stirring at room 10 hours; solvent evaporated, crystallization in the ethanol, thin layer is checked B ' 3 and panoxadiol; crystallization is dissolved in methyl alcohol; by silica gel column chromatography, 8: 2 wash-out panoxadiols of sherwood oil-acetone, again with chloroform-methanol wash at 9: 1 B ' 3; crystallization in vinyl acetic monomer, yield about 10%.