CN103193868B - A kind of purification process of echinocandin antifungal agent anidulafungin - Google Patents
A kind of purification process of echinocandin antifungal agent anidulafungin Download PDFInfo
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Abstract
The invention discloses a kind of purification process of echinocandin antifungal agent anidulafungin, comprise the following steps: (1) prepares dry sample: in anidulafungin crude product, add organic solvent dissolution, silica gel is added after abundant dissolving, dry after stirring, obtained anidulafungin dry sample; (2) pressurize wash-out: evenly fill out anidulafungin dry sample on the chromatography column top that silica gel is housed, add eluting solvent and carry out pressurization wash-out, monitor with high performance liquid chromatography, collect the elutriant that anidulafungin content is greater than 98%; (3) concentrated: anidulafungin content to be greater than the elutriant of 98%, to be concentrated into dry, obtain the anidulafungin sterling that content is greater than 98%.Present method adopts column chromatography, and easy and simple to handle, equipment cost is low; Select hypotoxicity, lower boiling organic solvent as elutriant, postorder recycling is simple, greatly reduces environmental protection pressure; The purifies and separates time is short, and purification effect is good, and purification yield is high, is suitable for carrying out industrialized production.
Description
Technical field
The present invention relates to a kind of purification process of echinocandin antifungal agent anidulafungin, belong to pharmaceutical synthesis technical field of refinement.
Background technology
In recent years, cause a large amount of uses of immunosuppressor, chemotherapy due to organ transfer operation and have more the reason such as abuse of the application of invasive therapy, the spreading of acquired immune deficiency syndrome (AIDS), extensive pedigree antibiotic, the crowd that immunity system is suppressed is on the increase, the sickness rate of fungi infestation significantly raises, and especially the sickness rate of deep fungal infection and lethality rate increase year by year.Meanwhile, along with the utilization of antifungal drug, the resistance of fungi is also more and more stronger, makes the application of antifungal drug have the swift and violent trend increased.Therefore, medicine one of study hotspot becoming anti-infective of anti-deep fungal infection, causes the concern of people day by day.
Echinocandin (echinocandins), also known as echinocandin, it is the newtype drug developed at the beginning of 21 century, there is the brand-new mechanism of action, anti-fungus spectra is comparatively wide, to Candida and Eurotium all effective, without cross resistance, without the obvious adverse reaction caused because of itself mechanism of action, it is the newtype drug being used for the treatment of systemic fungal infection at present.
Echinocandin belongs to acetyl six lopps, is glucan synthase inhibitors.Dextran is a kind of fungal cell wall polysaccharide, it is the important component of cell walls, cell walls can be made to keep integrity and make its osmotic pressure keep stable, echinocandin can suppress β-(1 in noncompetitive ground, 3) synthesis of-D-dextran, destroy the integrity of cell walls, and then play germicidal action.Due to the acellular wall of mammiferous cell, therefore this medicine is to human body fanout free region, and side effect is little, and safety coefficient is higher.
Anidulafungin (anidulafungin) is the semi-synthetic antifungal drug of third generation echinocandin class, is the derivative of echinocandin B, by ring six peptides produced from filamentous fungi A. nidulans---the side chain that ECB and synthesis obtain docks and forms.This medicine is developed by Vicuron drugmaker of the U.S., on May 24th, 2004 is by the clinical application of FDA, and in December, 2006 in U.S.'s Initial Public Offering, be applicable to following fungi infestation: the 1. monilial infection (peritoneal abscess, peritonitis) of candidemia and other types; 2. candidiasis of the esophagus.Because anidulafungin is removed without hepatic metabolism and kidney, drug combination and the bad patient of Liver and kidney function is made to become possibility without the need to adjusting dosage, and drug safety, better tolerance, has wide market outlook.
The molecular formula of anidulafungin is C
58h
73n
7o
17, molecular weight is 1140.24, and its structural formula is as follows:
Anidulafungin is a kind of white or off-white powder, be soluble in DMF, dimethyl sulfoxide (DMSO), dissolve in the mixed solvents such as methylene chloride-methanol, chloroform-methanol, acetonitrile-water, acetone-water, be slightly soluble in ethanol, methyl alcohol, tetrahydrofuran (THF), water insoluble.Abroad in patent WO9527074, US5541160, WO9508341, disclose the method using preparative high-performance liquid chromatographic anidulafungin to be carried out to purifying, adopt DELTA PAK C18 chromatographic column, acetonitrile/water solution is adopted to be eluent (also known as moving phase), gradient elution ratio is 40:60 to 55:45, and uv-absorbing wavelength is 210nm and 277nm.But the apparatus expensive that the method uses, processing sample is few, and the purifies and separates time is long, and aftertreatment is difficult, and cost of development is high, still cannot realize industrial amplification production at present.
Summary of the invention
The technical issues that need to address of the present invention are to provide that a kind of technique is simple, easy and simple to handle, solvent for use toxicity is low, product purity is high, be applicable to the purification process of the echinocandin antifungal agent anidulafungin of suitability for industrialized production.
For solving the problems of the technologies described above, the technical solution used in the present invention is:
A purification process for echinocandin antifungal agent anidulafungin, order is carried out according to the following steps:
(1) prepare dry sample: in echinocandin antifungal agent anidulafungin crude product, add organic solvent dissolution, after fully dissolving, add silica gel, dry after stirring, obtained anidulafungin silica gel dry sample;
(2) pressurize wash-out: evenly fill out anidulafungin silica gel dry sample on the chromatography column top that silica gel is housed, add eluting solvent and carry out pressurization wash-out, monitor with high performance liquid chromatography, collect the elutriant that anidulafungin content is greater than 98%;
(3) concentrated: elutriant anidulafungin content being greater than 98% is concentrated into dry, obtains the echinocandin antifungal agent anidulafungin sterling that content is greater than 98%.
Further improvement of the present invention is: the organic solvent used in described step (1) is methylene dichloride, the mixed solvent of any one or any two kinds in chloroform, tetrahydrofuran (THF), acetonitrile, acetone, methyl alcohol, ethanol.
Further improvement of the present invention is: in described step (1), the weight that adds of silica gel is anidulafungin crude product weight 1 ~ 3 times.
Further improvement of the present invention is: in described step (2), the aspect ratio of chromatography column is 8:1 ~ 18:1.
Further improvement of the present invention is: the silica gel weight loaded in the chromatography column of described step (2) is 20 ~ 30 times of anidulafungin crude product weight.
Further improvement of the present invention is: described step (1) and the middle silica gel used of step (2) are spherical silica gel or indefinite form silica gel, and silica gel granularity is 100 order ~ 400 orders.
Further improvement of the present invention is: the eluting solvent used in described step (2) is formed by two kinds of solvent; Wherein the first solvent is any one in methylene dichloride, chloroform, tetrahydrofuran (THF), acetonitrile, acetone, and the second solvent is any one in methyl alcohol, ethanol.
Further improvement of the present invention is: described pressurization wash-out is gradient pressurization wash-out, in the first solvent gradient used, the volume ratio of the first solvent and the second solvent is 90:10 ~ 80:20, and the volumetric usage of the first solvent gradient is 100 ~ 500 times of anidulafungin crude product weight; Volume ratio 80:20 ~ the 60:40 of the first solvent and the second solvent in the second solvent gradient used, the volumetric usage of the second solvent gradient is 20 ~ 100 times of anidulafungin crude product weight.
Further improvement of the present invention is: the post pressure of the pressurization wash-out in described step (2) is 2 ~ 8bar.
Further improvement of the present invention is: the concentrating under reduced pressure under 30 DEG C ~ 50 DEG C conditions of the simmer down in described step (3), is concentrated into no solvent residue.
Owing to have employed technique scheme, the technical progress acquired by the present invention is:
The invention provides a kind of purification process for echinocandin antifungal agent anidulafungin.Present method adopts common column chromatography to carry out purifying, and step is simple, and easy and simple to handle, equipment cost is low; Select hypotoxicity, lower boiling organic solvent as eluting solvent, postorder recycling is simple, greatly reduces environmental protection pressure; The purifies and separates time is short, and purification effect is good, and purification yield is high, is suitable for carrying out industrialized production.
The present invention selects column chromatography as purification process, different and be separated according to the adsorptive power of material on stationary phase.Stationary phase is loaded in post by column chromatography, and make sample to be separated reach separation along a direction reach, can be used for the material that segregation ratio is relatively large, separation accuracy is high, and equipment is simple, easy to operate, is a kind of purification process that can be used for large-scale industrial production.
The aspect ratio that present method limits chromatography column is 8:1 ~ 18:1, can ensure, on the basis that anidulafungin and other impurity are completely segregated, to shorten the post time, to meet industrialization production requirements as far as possible.
Through lot of experiment validation, present method selects silica gel as stationary phase, is filled in chromatography column, has than C18 reversed phase chromatography column packing and the more excellent separating effect of polystyrene/divinylbenzene microspheres.Silica gel has very high physical strength and good chemical stability, and easy to use, the life-span is long, and can repeatedly recycle, separating ranges is wide, and the separation especially for the organic compound of relative complex has higher efficiency.Wherein, the particle diameter of spherical silica gel is accurate, and post effect is relatively high; And indefinite form silica gel has larger specific surface area, high adsorption capacity, selectivity is stronger.
Present method uses a dry method on a sample, and adds silica gel adsorption, then solvent evaporate to dryness is obtained anidulafungin silica gel dry sample, evenly fill out on the chromatography column top that silica gel is housed, then carry out wash-out after fully being dissolved by anidulafungin crude product solvent.Although operate slightly loaded down with trivial details, be adsorbed on by product on silica gel and carry out wash-out again, solvent front easily flushes, and is conducive to impurity to be evenly separated.
Present method adopts gradient pressure wash-out, and compresses into the kind of eluting solvent, gradient concentration and post and gone restriction.Gradient elution is conducive to the separation of complicated components material, and its elutive power progressively increases, and separating power is strong, and analytical cycle shortens relatively, and can reduce conditions of streaking, can ensure anidulafungin sterling and separated from impurities, avoids intersecting.Use two kinds of organic solvents to be mixed to get eluting solvent, have the applicable polarity being conducive to being separated, and the organic solvent that present method is selected is hypotoxicity, lower boiling, postorder recycling is simple, greatly reduces environmental protection pressure.The post pressure of pressurization wash-out is defined as 2 ~ 8bar by present method, can obtain the high purity product that HPLC content is greater than 98%, and ensures higher yield.
Embodiment
Below in conjunction with embodiment, the present invention is described in further details:
Anidulafungin crude product used in the present invention is that North China Pharmacuetical Group New Drug Research & Development Co., Ltd produces; Indefinite form silica gel is that Qingdao Marine Chemical Co., Ltd. produces; Spherical silica gel is that Tianjin Beaune Ai Jieer scientific & technical corporation produces; C18 reversed phase chromatography column packing is that Tianjin Beaune Ai Jieer scientific & technical corporation produces; Polystyrene/divinylbenzene microspheres is that Suzhou Nano-Micro Bio-technology Co., Ltd. produces; High performance liquid chromatograph is that Waters company produces, 996 type detectors, 515 pumps; Other reagent are commercially available prod.
Embodiment 1
Get anidulafungin crude product 22.5g, add methyl alcohol and make it fully dissolve, then add 45g indefinite form silica gel (200 order ~ 300 order) and stir, after drying, obtain anidulafungin silica gel dry sample.
565g indefinite form silica gel (200 order ~ 300 order) is loaded in the chromatography column that aspect ratio is 8:1, anidulafungin silica gel dry sample is evenly filled out on chromatography column top, then gradient pressurization wash-out is carried out with methylene chloride-methanol mixing solutions as eluting solvent, in first solvent gradient, the volume ratio of methylene dichloride and methyl alcohol is 90:10, in second solvent gradient, the volume ratio of methylene dichloride and methyl alcohol is 80:20, and the post pressure in chromatography column is 5bar.According to the monitoring result of high performance liquid chromatography, collect the elutriant that anidulafungin content is greater than 98%.The first solvent gradient used and the consumption of the second solvent gradient are respectively 4500ml and 675ml.
Elutriant anidulafungin content being greater than 98% is evaporated to no solvent residue at 30 DEG C, obtains 14.1g anidulafungin sterling, yield 62.67%.
After testing, the HPLC content of the anidulafungin sterling of acquisition is 99.25%.
Embodiment 2
Get anidulafungin crude product 20.5g, add tetrahydrofuran (THF) and make it fully dissolve, then add 20.5g spherical silica gel (300 order ~ 400 order) and stir, after drying, obtain anidulafungin silica gel dry sample.
410g spherical silica gel (300 order ~ 400 order) is loaded in the chromatography column that aspect ratio is 9:1, anidulafungin silica gel dry sample is evenly filled out on chromatography column top, then gradient pressurization wash-out is carried out with tetrahydrofuran (THF)-methanol mixed solution as eluting solvent, in first solvent gradient, the volume ratio of tetrahydrofuran (THF) and methyl alcohol is 90:10, in second solvent gradient, the volume ratio of tetrahydrofuran (THF) and methyl alcohol is 75:25, and the post pressure in chromatography column is 6bar.According to the monitoring result of high performance liquid chromatography, collect the elutriant that anidulafungin content is greater than 98%.The first solvent gradient used and the consumption of the second solvent gradient are respectively 3050ml and 820ml.
Elutriant anidulafungin content being greater than 98% is evaporated to no solvent residue at 40 DEG C, obtains 12.2g anidulafungin sterling, yield 59.51%.
After testing, the HPLC content of the anidulafungin sterling of acquisition is 98.60%.
Embodiment 3
Get anidulafungin crude product 15.2g, add methylene chloride-methanol and make it fully dissolve, then add 45.6g indefinite form silica gel (100 order ~ 200 order) and stir, after drying, obtain anidulafungin silica gel dry sample.
456g indefinite form silica gel (100 order ~ 200 order) is loaded in the chromatography column that aspect ratio is 12:1, anidulafungin silica gel dry sample is evenly filled out on chromatography column top, then gradient pressurization wash-out is carried out with chloroform-methanol mixing solutions as eluting solvent, in first solvent gradient, the volume ratio of chloroform and methyl alcohol is 90:10, in second solvent gradient, the volume ratio of chloroform and methyl alcohol is 70:30, and the post pressure in chromatography column is 2bar.According to the monitoring result of high performance liquid chromatography, collect the elutriant that anidulafungin content is greater than 98%.The first solvent gradient used and the consumption of the second solvent gradient are respectively 2280ml and 710ml.
Elutriant anidulafungin content being greater than 98% is evaporated to no solvent residue at 35 DEG C, obtains 8.0g anidulafungin sterling, yield 52.63%.
After testing, the HPLC content of the anidulafungin sterling of acquisition is 98.22%.
Embodiment 4
Get anidulafungin crude product 25.3g, add chloroform-acetonitrile and make it fully dissolve, then add 38g indefinite form silica gel (200 order ~ 300 order) and stir, after drying, obtain anidulafungin silica gel dry sample.
630g indefinite form silica gel (200 order ~ 300 order) is loaded in the chromatography column that aspect ratio is 14:1, anidulafungin silica gel dry sample is evenly filled out on chromatography column top, then gradient pressurization wash-out is carried out with dichloromethane-ethanol mixing solutions as eluting solvent, in first solvent gradient, the volume ratio of methylene dichloride and ethanol is 90:10, in second solvent gradient, the volume ratio of methylene dichloride and ethanol is 75:25, and the post pressure in chromatography column is 4bar.According to the monitoring result of high performance liquid chromatography, collect the elutriant that anidulafungin content is greater than 98%.The first solvent gradient used and the consumption of the second solvent gradient are respectively 6300ml and 2000ml.
Elutriant anidulafungin content being greater than 98% is evaporated to no solvent residue at 50 DEG C, obtains 13.3g anidulafungin sterling, yield 52.57%.
After testing, the HPLC content of the anidulafungin sterling of acquisition is 98.62%.
Embodiment 5
Get anidulafungin crude product 13.5g, add tetrahydrofuran (THF)-methyl alcohol and make it fully dissolve, then add 33.75g indefinite form silica gel (300 order ~ 400 order) and stir, after drying, obtain anidulafungin silica gel dry sample.
370g indefinite form silica gel (300 order ~ 400 order) is loaded in the chromatography column that aspect ratio is 18:1, anidulafungin silica gel dry sample is evenly filled out on chromatography column top, then gradient pressurization wash-out is carried out with tetrahydrofuran-ethyl alcohol mixing solutions as eluting solvent, in first solvent gradient, the volume ratio of tetrahydrofuran (THF) and ethanol is 85:15, in second solvent gradient, the volume ratio of tetrahydrofuran (THF) and ethanol is 70:30, and the post pressure in chromatography column is 8bar.According to the monitoring result of high performance liquid chromatography, collect the elutriant that anidulafungin content is greater than 98%.The first solvent gradient used and the consumption of the second solvent gradient are respectively 4050ml and 950ml.
Elutriant anidulafungin content being greater than 98% is evaporated to no solvent residue at 35 DEG C, obtains 6.6g anidulafungin sterling, yield 48.89%.
After testing, the HPLC content of the anidulafungin sterling of acquisition is 99.17%.
Embodiment 6
Get anidulafungin crude product 20.0g, add acetone-methanol and make it fully dissolve, then add 25g spherical silica gel (100 order ~ 300 order) and stir, after drying, obtain anidulafungin silica gel dry sample.
440g spherical silica gel (100 order ~ 300 order) is loaded in the chromatography column that aspect ratio is 8:1, anidulafungin silica gel dry sample is evenly filled out on chromatography column top, then gradient pressurization wash-out is carried out with acetonitrile-ethanol mixing solutions as eluting solvent, in first solvent gradient, the volume ratio of acetonitrile and ethanol is 80:20, in second solvent gradient, the volume ratio of acetonitrile and ethanol is 70:30, and the post pressure in chromatography column is 3bar.According to the monitoring result of high performance liquid chromatography, collect the elutriant that anidulafungin content is greater than 98%.The first solvent gradient used and the consumption of the second solvent gradient are respectively 7000ml and 400ml.
Elutriant anidulafungin content being greater than 98% is evaporated to no solvent residue at 45 DEG C, obtains 8.5g anidulafungin sterling, yield 42.50%.
After testing, the HPLC content of the anidulafungin sterling of acquisition is 98.53%.
Embodiment 7
Get anidulafungin crude product 12.6g, add chloroform and make it fully dissolve, then add 22g indefinite form silica gel (100 order ~ 400 order) and stir, after drying, obtain anidulafungin silica gel dry sample.
340g indefinite form silica gel (100 order ~ 400 order) is loaded in the chromatography column that aspect ratio is 17:1, anidulafungin silica gel dry sample is evenly filled out on chromatography column top, then gradient pressurization wash-out is carried out with acetone-methanol mixing solutions as eluting solvent, in first solvent gradient, the volume ratio of acetone and methyl alcohol is 80:20, in second solvent gradient, the volume ratio of acetone and methyl alcohol is 65:35, and the post pressure in chromatography column is 7bar.According to the monitoring result of high performance liquid chromatography, collect the elutriant that anidulafungin content is greater than 98%.The first solvent gradient used and the consumption of the second solvent gradient are respectively 8040ml and 1260ml.
Elutriant anidulafungin content being greater than 98% is evaporated to no solvent residue at 30 DEG C, obtains 6.2g anidulafungin sterling, yield 49.21%.
After testing, the HPLC content of the anidulafungin sterling of acquisition is 98.39%.
Embodiment 8
Get anidulafungin crude product 22.7g, add chloroform-ethanol and make it fully dissolve, then add 51g spherical silica gel (200 order ~ 400 order) and stir, after drying, obtain anidulafungin silica gel dry sample.
681g spherical silica gel (200 order ~ 400 order) is loaded in the chromatography column that aspect ratio is 15:1, anidulafungin silica gel dry sample is evenly filled out on chromatography column top, then gradient pressurization wash-out is carried out with chloroform-ethanol mixing solutions as eluting solvent, in first solvent gradient, the volume ratio of chloroform and ethanol is 82:18, in second solvent gradient, the volume ratio of chloroform and ethanol is 60:40, and the post pressure in chromatography column is 8bar.According to the monitoring result of high performance liquid chromatography, collect the elutriant that anidulafungin content is greater than 98%.The first solvent gradient used and the consumption of the second solvent gradient are respectively 6000ml and 1200ml.
Elutriant anidulafungin content being greater than 98% is evaporated to no solvent residue at 50 DEG C, obtains 13.8g anidulafungin sterling, yield 60.79%.
After testing, the HPLC content of the anidulafungin sterling of acquisition is 99.07%.
Embodiment 9
Get anidulafungin crude product 15.5g, add methylene dichloride and make it fully dissolve, then add 44.5g spherical silica gel (100 order ~ 200 order) and stir, after drying, obtain anidulafungin silica gel dry sample.
434g spherical silica gel (100 order ~ 200 order) is loaded in the chromatography column that aspect ratio is 10:1, anidulafungin silica gel dry sample is evenly filled out on chromatography column top, then gradient pressurization wash-out is carried out with tetrahydrofuran-ethyl alcohol mixing solutions as eluting solvent, in first solvent gradient, the volume ratio of tetrahydrofuran (THF) and ethanol is 85:15, in second solvent gradient, the volume ratio of tetrahydrofuran (THF) and ethanol is 60:40, and the post pressure in chromatography column is 2bar.According to the monitoring result of high performance liquid chromatography, collect the elutriant that anidulafungin content is greater than 98%.The first solvent gradient used and the consumption of the second solvent gradient are respectively 7750ml and 1500ml.
Elutriant anidulafungin content being greater than 98% is evaporated to no solvent residue at 40 DEG C, obtains 8.9g anidulafungin sterling, yield 57.42%.
After testing, the HPLC content of the anidulafungin sterling of acquisition is 98.05%.
In above-described embodiment 1 ~ embodiment 9, according to the difference of actual elute effect, the first solvent gradient used and the consumption of the second solvent gradient can change to some extent.On the whole, the volumetric usage of the first solvent gradient is 100 ~ 500 times of anidulafungin crude product weight, and the volumetric usage of the second solvent gradient is 20 ~ 100 times of anidulafungin crude product weight.
Embodiment 10
The post pressure that the present embodiment is used for when verifying pressurization wash-out is on the impact of purifies and separates efficiency and purification yield, and except post pressure, the charging capacity of anidulafungin crude product is all identical with embodiment 1 with other all purification conditions.When post pressure is respectively 0bar, 2bar, 4bar, 6bar, 8bar and 10bar, its purifies and separates efficiency is in table 1.
The different post pressure of table 1 is on the impact of column chromatography separating effect
Can be found out by data in table 1, when post pressure is 0bar, when namely adopting normal pressure wash-out, disengaging time reaches 60h, still cannot obtain the anidulafungin elutriant that content is greater than 98%; And when post pressure is for 10bar, because post pressure increases, elution speed is accelerated, cause anidulafungin and magazins' layout not thorough, the HPLC content of products obtained therefrom is low, cannot meet the requirement of content of drug, and purification yield is very low.Therefore, present method determines that the post pressure of pressurization wash-out is 2 ~ 8bar, carries out gradient pressurization wash-out, can obtain the high purity product that HPLC content is greater than 98%, and ensure higher yield under the post press strip part of this scope.
Embodiment 11
The present embodiment is used for verifying the impact using different stationary phase on purifies and separates efficiency and purification yield, and except stationary phase, anidulafungin crude product charging capacity is all identical with embodiment 1 with other all purification conditions.When stationary phase is indefinite form silica gel (normal phase column filler), spherical silica gel (normal phase column filler), C18 reversed phase chromatography column packing, polystyrene and divinylbenzene microspheres respectively, its purifies and separates efficiency is in table 2.
The different stationary phase of table 2 is on the impact of column chromatography separating effect
Can be found out by data in table 2, when using indefinite form silica gel and spherical silica gel as stationary phase, products obtained therefrom all far away higher than the related data of other two kinds of filler products obtained therefroms, obviously has more superior separating effect in HPLC content and yield two.
When using the C18 reversed-phase column filler of 30 ~ 50 μm as stationary phase, not only the time of purifies and separates extends greatly, and purification yield is on the low side; When using polystyrene/divinylbenzene microspheres as stationary phase, obtain the HPLC content of product lower than 98%, and purification yield is only 15.11%, and these two kinds of stationary phase are all not suitable for the requirement of industrialized production.
Claims (5)
1. a purification process for echinocandin antifungal agent anidulafungin, is characterized in that order is carried out according to the following steps:
(1) prepare dry sample: in echinocandin antifungal agent anidulafungin crude product, add organic solvent dissolution, after fully dissolving, add silica gel, dry after stirring, obtained anidulafungin silica gel dry sample;
(2) pressurize wash-out: evenly fill out anidulafungin silica gel dry sample on the chromatography column top that silica gel is housed, add eluting solvent and carry out pressurization wash-out, monitor with high performance liquid chromatography, collect the elutriant that anidulafungin content is greater than 98%;
(3) concentrated: elutriant anidulafungin content being greater than 98% is concentrated into dry, obtains the echinocandin antifungal agent anidulafungin sterling that content is greater than 98%;
The eluting solvent used in described step (2) is formed by two kinds of solvent; Wherein, the first solvent is any one in methylene dichloride, chloroform, tetrahydrofuran (THF), acetonitrile, acetone, and the second solvent is any one in methyl alcohol, ethanol;
Described pressurization wash-out is gradient pressurization wash-out, and the post pressure of pressurization wash-out is 2 ~ 8bar; In the first solvent gradient used, the volume ratio of the first solvent and the second solvent is 90:10 ~ 80:20, and the volumetric usage of the first solvent gradient is 100 ~ 500 times of anidulafungin crude product weight; Volume ratio 80:20 ~ the 60:40 of the first solvent and the second solvent in the second solvent gradient used, the volumetric usage of the second solvent gradient is 20 ~ 100 times of anidulafungin crude product weight;
The silica gel weight loaded in the chromatography column of described step (2) is 20 ~ 30 times of anidulafungin crude product weight;
Described step (1) and the middle silica gel used of step (2) are spherical silica gel or indefinite form silica gel, and silica gel granularity is 100 order ~ 400 orders.
2. the purification process of a kind of echinocandin antifungal agent anidulafungin according to claim 1, is characterized in that: the organic solvent used in described step (1) is methylene dichloride, the mixed solvent of any one or any two kinds in chloroform, tetrahydrofuran (THF), acetonitrile, acetone, methyl alcohol, ethanol.
3. the purification process of a kind of echinocandin antifungal agent anidulafungin according to claim 1, is characterized in that: in described step (1), silica gel adds 1 ~ 3 times that weight is anidulafungin crude product weight.
4. the purification process of a kind of echinocandin antifungal agent anidulafungin according to claim 1, is characterized in that: in described step (2), the aspect ratio of chromatography column is 8:1 ~ 18:1.
5. the purification process of a kind of echinocandin antifungal agent anidulafungin according to claim 1, is characterized in that: the concentrating under reduced pressure under 30 DEG C ~ 50 DEG C conditions of the simmer down in described step (3), is concentrated into no solvent residue.
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| CN104447961B (en) * | 2013-09-13 | 2018-05-15 | 浙江震元制药有限公司 | The extracting method of echinocandin B parent nucleus |
| CN103555591B (en) * | 2013-10-12 | 2015-06-03 | 浙江工业大学 | Method and bacterial strain for fermentation preparation of Echinocandin B |
| CN103965298B (en) * | 2014-05-23 | 2016-08-10 | 浙江海正药业股份有限公司 | A kind of purification process of anidulafungin |
| CN104130314A (en) * | 2014-07-15 | 2014-11-05 | 华北制药集团新药研究开发有限责任公司 | Anidulafungin refining method |
| CN106860856B (en) * | 2015-12-14 | 2019-05-31 | 山东新时代药业有限公司 | A kind of freeze-dried powder and preparation method containing anidulafungin |
| CN112415156A (en) * | 2019-08-22 | 2021-02-26 | 华北制药集团新药研究开发有限责任公司 | Method for detecting bacterial endotoxin of anidulafungin raw material medicine |
| CN112430256A (en) * | 2020-11-23 | 2021-03-02 | 苏州纳微科技股份有限公司 | Method for separating and purifying anidulafungin mother nucleus |
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