CN102219764B - Method for separating and purifying paclitaxel industrially - Google Patents
Method for separating and purifying paclitaxel industrially Download PDFInfo
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- DBXFAPJCZABTDR-WBYYIXQISA-N cephalomannine Chemical compound O([C@@H]1[C@]2(O)C[C@@H](C(=C([C@@H](OC(C)=O)C(=O)[C@]3(C)[C@@H](O)C[C@H]4OC[C@]4([C@H]31)OC(C)=O)C2(C)C)C)OC(=O)[C@H](O)[C@@H](NC(=O)C(/C)=C/C)C=1C=CC=CC=1)C(=O)C1=CC=CC=C1 DBXFAPJCZABTDR-WBYYIXQISA-N 0.000 claims abstract description 21
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Abstract
The invention discloses a method for separating and purifying paclitaxel industrially and efficiently, and belongs to the field of the separation and purification of compounds. In the method, the paclitaxel is separated by column chromatography, namely the paclitaxel is separated and purified by taking a combination of any three or four of compounds of alkane having 1 to 10 carbon atoms, chlorohydrocarbons, alcohols having 1 to 4 carbon atoms, saturated alkane esters and alkane ketones, and amine compounds as a mobile phase and 200 to 300-mesh ordinary silica gel as a stationary phase by normal phase column chromatography. By selecting a multielement system mixed solvent as the mobile phase, the separation degree of substances which are difficult to separate is increased, and the problem that the paclitaxel cannot be separated from cephalomannine effectively in the conventional separation method is solved. After the paclitaxel is separated and purified by the column chromatography, the purity of paclitaxel crude products serving as raw materials can be improved to be over 98 percent, and according to the condition of the raw materials, the total yield is up to 90 percent only by eluting the paclitaxel crude products once to twice. In the method, the common solvent and silica gel are used, so the cost is low, the method is easy and convenient to operate, the sample volume is large, and industrial production is easy to realize.
Description
Technical field
The present invention relates to the separation purification method of taxol, be specifically related to a kind of method of industrial high efficiency separating and purifying taxol and Cephalomannine, belong to compound separation purifying field.
Background technology
Taxol is a kind of new type anticancer medicine with Taxan diterpene skeleton of separation and Extraction from Chinese yew genus plants, is also unique a kind of microtubule polymerization and stable medicine of polymerization microtubule of promoting of understanding at present.It has powerful restraining effect to most of solid tumors, and on normal cell based this without impact, especially less to determined curative effect, the side effect of advanced ovarian cancer, mammary cancer, nonsmall-cell lung cancer and Kaposis sarcoma.Along with further going deep into of research, the antitumous effect of taxanes medicine more and more is more widely used, and has now become the antitumor drug of a line cancer therapy drug and wide spectrum.
Ramulus et folium taxi cuspidatae class plant as the taxol main source belongs to rareness species, is listed in the first-grade state protection plant in China.In bark of Ramulus et folium taxi cuspidatae, content of taxol is extremely low, accounts for 0.01%, far away can not satisfying the market to the demand of taxol.Over nearly 20 years, the preparation method of taxol has multidisciplinary in succession, with multi-angle the exploration report.Taxol cell cultures, fungi fermentation, molecular design and total synthesis method are arranged.Where no matter take the standby taxol of legal system, all need taxol is separated also purifying out from paclitaxel analogs.Common paclitaxel analogs mainly contains Cephalomannine, baccatin III, 10-deacetylate taxol, 7-epi-taxol, taxol C etc.Because structure is more close with character, thereby cause taxol very difficult with separating of its analogue.In addition, as a kind of biomacromolecule material, taxol is subjected to the impact of the envrionment conditionss such as temperature, organic solvent, acid, alkali, degraded or isomery easily occurs and generate other taxane substances.For example taxol can be degraded to baccatin III or isomery occurs and generate 7-epi-taxol under strongly-acid or weak basic condition; When temperature is higher, DeR also can occur in taxol, generates corresponding small-molecule substance.As the medicine in clinical use, taxol its purity requirement in actual applications is very high.The taxol resource is precious, hold at high price, and the purity of separation and yield become the key factor of restriction pharmaceutical paclitaxel cost.Based on above-mentioned several aspects, make more efficient, the cheap industrial separation purification technique of exploitation and novel production separation purifying technique, become the focal issue in taxol research.
Separating and purifying taxol commonly used and the method for paclitaxel analogs mainly contain column chromatography, tlc, the precipitator method, miccellar electrokinetic chromatography method, membrane separation process, resin absorption partition method, chemical reaction method etc. at present.But what be applicable to large-scale industrialization production only has column chromatography a kind of, and additive method all is confined to laboratory study, and column chromatography is divided into again normal phase column method and two kinds of methods of reversed-phase column.
The patent application of relevant Purification of Taxol has multinomial: as Chinese patent CN 1377882A " a kind of preparation high-purity taxol, Cephalomannine, method of 10-deacetylation Bakating III of uniting "; Chinese patent CN 1611496A " a kind of method of utilizing high pressure liquid chromatography to prepare high-purity taxol "; Chinese patent CN 101391989A " a kind of method for preparing polyhydroxy taxone and taxol ".
The weak point of existing Purification of Taxol method mainly is: the method for present various isolation of taxol is isolation of taxol and Cephalomannine effectively still; With regard to can the heavy industrialization separating and purifying taxol and the column chromatography of paclitaxel analogs with regard to, there are again defective separately in normal phase method and reversed phase method.The normal phase column chromatography method often needs to introduce oxidizing reaction, by changing the chemical structure of Cephalomannine, to increase the resolution of Cephalomannine and taxol, this has not only changed the chemical structure of separate object, and to repeatedly cross post, repeatedly recrystallization carries out purifying.The loaded down with trivial details yield that not only reduces the target taxol of operation steps increases production cost, and separating resulting only can obtain a kind of product of taxol, and the Cephalomannine after change structure can only be abandoned as waste material, has not only wasted resource but also contaminate environment.Although the reversed phase column chromatography method need not change the chemical structure of separate object, in practical application, the method need to be used special reversed-phase column silica gel, and its market value is high, reaches 500,000 yuan/kilogram.In suitability for industrialized production, 1 kilogram of taxol of every separation can be used the reversed-phase column silica gel of 50 kilograms at least, and only the cost of one, reversed-phase column silica gel will 2,500 ten thousand yuan, has greatly increased the production cost of isolation of taxol, has limited its large-scale industrialization and has produced.
Summary of the invention
The object of the present invention is to provide a kind of Purification of Taxol method that separation efficiency is high, with low cost, be easy to the industrialization operation.
For realizing the object of the invention, adopt the normal phase column chromatography method, polynary by selecting (quaternary or five yuan) system mixed solvent carries out separation and purification as moving phase to the thick product of taxol.Moving phase is the combination of any three kinds or four kinds and aminated compounds in C1-10 alkanes, chlorinated hydrocarbons, C1-4 alcohols, saturated alkane ester class, alkane ketone compounds.
Described amine solvent is selected from diethylamine or triethylamine; The C1-10 alkane solvents is selected from normal hexane, Skellysolve A or hexanaphthene; The chlorinated hydrocarbons solvent is selected from methylene dichloride, trichloromethane or tetrachloromethane; Described C1-4 alcoholic solvent is selected from methyl alcohol or ethanol; Described saturated alkane esters solvent is selected from ethyl acetate; Described alkane ketone kind solvent is selected from acetone or isopropyl acetone.The consumption of amine solvent accounts for 8%～16% of mobile phase volume.
Usually through one to the twice chromatographic post wash-out the taxol of purity 93%～95% account for 80%～90%; Through one to the twice chromatographic post wash-out the taxol of purity 30%～90% account for 5%～15%; Through one to the twice chromatographic post wash-out purity account for 1%～3% lower than 30% taxol; Sample carries out recrystallization one time, gets the target taxol.
In practical application, the content of raw material taxol is higher, and the wash-out number of times of required chromatography column is fewer.When adopting present method isolation of taxol, higher than 70%, only need a chromatography column wash-out just can obtain desirable separating effect as the raw material content of taxol; Be 50%～70% as the raw material content of taxol, twice chromatographic post wash-out just can obtain desirable separating effect.
The selection of system moving phase that innovative point of the present invention is is polynary (quaternary or five yuan), make chemical reaction step such as having saved bromination method in the separation and purification process, avoided the change of chemical structure in the separate object, reduced the post number of times excessively of material sample, only crossed a chromatography column and just the taxol more than 80% can be separated.Simultaneously, separating resulting obtains highly purified taxol and two kinds of materials of Cephalomannine.Cephalomannine also can be made into Docetaxel or taxol except the bulk drug that itself can be used as high added value directly utilizes (150,000 yuan/kilogram), obtain more economic benefit.
Innovative point of the present invention is that moving phase selects the combination of any three kinds or four kinds and aminated compounds in C1-10 alkanes, chlorinated hydrocarbons, C1-4 alcohols, saturated alkane ester class, alkane ketone compounds; Stationary phase is common 200 orders～300 order normal phase column silica gel of 50 yuan/Kg.In the separation and purification process material therefor be simple and easy to, material with low cost, be easy to carry out suitability for industrialized production and reduce production costs.Compare with reversed-phase column isolation of taxol method commonly used at present, effectively isolation of taxol and Cephalomannine, cross the post number of times few, and separation efficiency is high, and low production cost can be carried out suitability for industrialized production on a large scale.
Innovative point of the present invention is the application of amine reagent, regulated the pH value of moving phase, sepn process is carried out under alkaline condition, this moment, separated material all existed with molecular form, reduced the absorption on silica gel, and improved difficult separate substance to the resolution of taxol and Cephalomannine, solved well in various separation methods in the past the effectively difficult problem of isolation of taxol and Cephalomannine.
Innovative point of the present invention is that in the elutriant of chromatography column, the liquid that contains Cephalomannine first flows out, and flows out after containing the liquid of taxol.This use with polynary (quaternary or five yuan) system moving phase and amine reagent is relevant, and alkaline isolating environment makes Cephalomannine flow out prior to taxol.
Adopt the inventive method effectively isolation of taxol and Cephalomannine, separation and purification is effective, can easily the thick product purity of taxol be increased to more than 98%, according to raw material condition, only once can be up to 90% to twice wash-out total recovery.With existing taxol separation method ratio, it is few, easy and simple to handle that present method is crossed post wash-out number of times, safety, and separation efficiency is high, and is with low cost, and applied sample amount is large, is easy to industrialization and carries out scale operation.
Embodiment
For the present invention is illustrated better, embodiment is as follows:
Embodiment 1
The sample of getting content of taxol 70% is raw material, and raw material and silica gel weight ratio are 2:100.Raw material is rear standby with the methylene dichloride dissolving.Quaternary system moving phase is normal hexane: methylene dichloride: methyl alcohol: triethylamine volume ratio=3:6:2:2,200 orders～300 order silica gel are as stationary phase.Adopt the wet method upper prop, adopt the wet method upper prop, after loading, with separating with the moving phase for preparing again after column volume of dichloromethane solvent punching.After 1.4 column volumes of moving phase punching, begin to have taxol C to flow out, and then flow out for Cephalomannine, after 0.3 column volume, for single taxol flows out, then taxol can be separated fully after 2 column volumes, and after a wash-out, the taxol of purity more than 90% accounts for 85%.Merge the taxol effluent liquid of purity 30%～90% and carry out the secondary wash-out.(sepn process can be monitored with TLC or HPLC) merges the taxol effluent liquid of twice elutriant moderate purity more than 90%, carry out vacuum concentration under 45 ℃, enriched material at room temperature (25 ℃) is used acetone: normal hexane=1:6 volume ratio is carried out recrystallization, there are a large amount of white precipitates to separate out after stirring 10min, after stirring 20min, taxol can be separated out fully.After carrying out suction filtration and vacuum-drying, obtain purity and reach 98.6% pure product of paclitaxel, the taxol total recovery reaches 97.6%.
Embodiment 2
The sample of getting content of taxol 60% is raw material, and raw material and silica gel weight ratio are 1.5:100.Raw material is rear standby with the methylene dichloride dissolving.Quinary system moving phase is normal hexane: methylene dichloride: acetone: triethylamine: ethyl acetate volume ratio=6:3:1:1:1.5,200 orders～300 order silica gel are as stationary phase.Adopt the wet method upper prop, after loading, with separating with the moving phase for preparing again after column volume of dichloromethane solvent punching.After 1.7 column volumes of moving phase punching, begin to have taxol C to flow out, and then flow out for Cephalomannine, after 0.5 column volume, for single taxol flows out, then taxol can be separated fully after 2 column volumes, and after a wash-out, the taxol of purity more than 90% accounts for 75%.Merge the taxol effluent liquid of purity 30%～90% and carry out the secondary wash-out.(sepn process can be monitored with TLC or HPLC) merges the taxol effluent liquid of twice elutriant moderate purity more than 90%, the effluent liquid that (sepn process can be monitored with TLC or HPLC) contains taxol carries out vacuum concentration under 45 ℃, enriched material at room temperature (25 ℃) is used acetone: normal hexane=1:6 volume ratio is carried out recrystallization, there are a large amount of white precipitates to separate out after stirring 10min, after stirring 20min, taxol can be separated out fully.After carrying out suction filtration and vacuum-drying, obtain purity and reach 98.5% pure product of paclitaxel, the taxol total recovery reaches 94.6%.
Embodiment 3
The sample of getting content of taxol 50% is raw material, and raw material and silica gel weight ratio are 2:100.Raw material is rear standby with the trichloromethane dissolving.Quinary system moving phase is normal hexane: trichloromethane: acetone: diethylamine: ethyl acetate volume ratio=4:3:1:1:1.5,200 orders～300 order silica gel are as stationary phase.Adopt the wet method upper prop, after loading, with separating with the moving phase for preparing again after 2 column volumes of trichloromethane solvent punching.After 2 column volumes of moving phase punching, begin to have taxol C to flow out, and then flow out for Cephalomannine, after 0.5 column volume, for single taxol flows out, then taxol can be separated fully after 2.5 column volumes, and after a wash-out, the taxol of purity more than 90% accounts for 66.7%.Merge the taxol effluent liquid of purity 30%～90% and carry out the secondary wash-out.The effluent liquid that (sepn process can be monitored with TLC or HPLC) contains taxol carries out vacuum concentration under 45 ℃, enriched material at room temperature (25 ℃) is used acetone: normal hexane=1:6 volume ratio is carried out recrystallization, there are a large amount of white precipitates to separate out after stirring 10min, after stirring 20min, taxol can be separated out fully, after carrying out suction filtration and vacuum-drying, obtain purity and reach 98.3% pure product of paclitaxel, the taxol total recovery reaches 90.7%.
Claims (2)
1. the method for an industrial separation purification of paclitaxel, adopt normal phase column chromatography method isolation of taxol, it is characterized in that, the sample of getting content of taxol 60% is raw material, and raw material and silica gel weight ratio are 1.5:100, and raw material is rear standby with the methylene dichloride dissolving; Quinary system moving phase is normal hexane: methylene dichloride: acetone: triethylamine: ethyl acetate volume ratio=6:3:1:1:1.5, and 200 orders～300 order silica gel are as stationary phase; Adopt the wet method upper prop, after loading, with separating with the moving phase for preparing again after column volume of dichloromethane solvent punching; After 1.7 column volumes of moving phase punching, begin to have taxol C to flow out, and then flow out for Cephalomannine, after 0.5 column volume, for single taxol flows out, taxol is separated fully after 2 column volumes again, and after a wash-out, the taxol of purity more than 90% accounts for 75%, merges the taxol effluent liquid of purity 30%～90% and carries out the secondary wash-out; Merge the taxol effluent liquid of twice elutriant moderate purity more than 90%, the effluent liquid that contains taxol carries out vacuum concentration under 45 ℃, enriched material is used acetone under 25 ℃ of room temperatures: normal hexane=1:6 volume ratio is carried out recrystallization, there are a large amount of white precipitates to separate out after stirring 10min, after stirring 20min, taxol is separated out fully; After carrying out suction filtration and vacuum-drying, obtain purity and reach 98.5% pure product of paclitaxel, the taxol total recovery reaches 94.6%.
2. the method for an industrial separation purification of paclitaxel, adopt normal phase column chromatography method isolation of taxol, it is characterized in that,
The sample of getting content of taxol 50% is raw material, and raw material and silica gel weight ratio are 2:100, and raw material is rear standby with the trichloromethane dissolving; Quinary system moving phase is normal hexane: trichloromethane: acetone: diethylamine: ethyl acetate volume ratio=4:3:1:1:1.5,200 orders～300 order silica gel are as stationary phase, adopt the wet method upper prop, after loading, with separating with the moving phase for preparing again after 2 column volumes of trichloromethane solvent punching; After 2 column volumes of moving phase punching, begin to have taxol C to flow out, and then flow out for Cephalomannine, after 0.5 column volume, for single taxol flows out, then taxol is separated fully after 2.5 column volumes, and after a wash-out, the taxol of purity more than 90% accounts for 66.7%; Merge the taxol effluent liquid of purity 30%～90% and carry out the secondary wash-out, the effluent liquid that contains taxol carries out vacuum concentration under 45 ℃, enriched material is used acetone under 25 ℃ of room temperatures: normal hexane=1:6 volume ratio is carried out recrystallization, there are a large amount of white precipitates to separate out after stirring 10min, after stirring 20min, taxol is separated out fully, after carrying out suction filtration and vacuum-drying, obtain purity and reach 98.3% pure product of paclitaxel, the taxol total recovery reaches 90.7%.
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| CN102417492A (en) * | 2011-12-07 | 2012-04-18 | 福建紫杉园生物有限公司 | Method for separating and purifying paclitaxel |
| CN103232416B (en) * | 2013-04-22 | 2015-03-18 | 无锡尔云科技有限公司 | Method for separating and purifying 10-deacetylated paclitaxel (10-DAP) |
| CN105457337A (en) * | 2015-11-17 | 2016-04-06 | 重庆臻源红豆杉发展有限公司 | A silica gel loading and sample loading method of a chromatographic column |
| CN109384749A (en) * | 2018-12-26 | 2019-02-26 | 重庆市碚圣医药科技股份有限公司 | A kind of purification process of taxol |
| CN112645906A (en) * | 2020-12-29 | 2021-04-13 | 重庆臻源红豆杉发展有限公司 | Method for separating and purifying paclitaxel by high-efficiency chromatography |
| CN113444060A (en) * | 2021-08-09 | 2021-09-28 | 怀化市盛德生物科技有限责任公司 | Paclitaxel separation and purification process |
| CN113717131B (en) * | 2021-08-27 | 2023-09-22 | 常熟纳微生物科技有限公司 | Separation and purification method of paclitaxel |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1069965A (en) * | 1991-04-19 | 1993-03-17 | 密西西比大学 | Separate the method for derivative of taxane and contain its composition |
| CN1377882A (en) * | 2001-04-02 | 2002-11-06 | 北京普瑞孚天然药物现代纯化和分离研究所有限公司 | Process for coproducing high purity taxol cephalomtanine, and 10-deacetyl Bakating III |
| CN1396917A (en) * | 2000-11-28 | 2003-02-12 | 因德纳有限公司 | Chromatographic separation method of paclitaxel and cephalomannin |
| CN1791590A (en) * | 2003-03-17 | 2006-06-21 | 天然医药品公司 | Purification of taxanes and taxane mixtures using polyethyleneimine-bonded resins |
-
2011
- 2011-05-12 CN CN 201110121650 patent/CN102219764B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1069965A (en) * | 1991-04-19 | 1993-03-17 | 密西西比大学 | Separate the method for derivative of taxane and contain its composition |
| CN1396917A (en) * | 2000-11-28 | 2003-02-12 | 因德纳有限公司 | Chromatographic separation method of paclitaxel and cephalomannin |
| CN1377882A (en) * | 2001-04-02 | 2002-11-06 | 北京普瑞孚天然药物现代纯化和分离研究所有限公司 | Process for coproducing high purity taxol cephalomtanine, and 10-deacetyl Bakating III |
| CN1791590A (en) * | 2003-03-17 | 2006-06-21 | 天然医药品公司 | Purification of taxanes and taxane mixtures using polyethyleneimine-bonded resins |
Non-Patent Citations (1)
| Title |
|---|
| 雒丽娜.紫杉醇和三尖杉宁碱的分离纯化及检测方法研究.《北京化工大学硕士研究生学位论文》.2005,摘要第V页第2段，正文第54页第4.4.1部分和表4-1，正文第55页倒数第3段和倒数第2段. * |
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