CN102464668B - Preparative chromatography purification method for purifying rapamycin or derivative thereof - Google Patents
Preparative chromatography purification method for purifying rapamycin or derivative thereof Download PDFInfo
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- CN102464668B CN102464668B CN201010546460.6A CN201010546460A CN102464668B CN 102464668 B CN102464668 B CN 102464668B CN 201010546460 A CN201010546460 A CN 201010546460A CN 102464668 B CN102464668 B CN 102464668B
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- 0 C[C@](CC(CC[C@]1OC(C(C)(C*)CO)=O)C[C@]1OC)C(CC([C@](C)C=C(C)[C@]([C@](C([C@](C)CCC=CC=CC=C(C)[C@@](CC(CC[C@]1C)O[C@]1(C(C(C1[C@]2CCCC1)=O)=O)N=O)OC)=O)OC)N=O)=O)OC2=O Chemical compound C[C@](CC(CC[C@]1OC(C(C)(C*)CO)=O)C[C@]1OC)C(CC([C@](C)C=C(C)[C@]([C@](C([C@](C)CCC=CC=CC=C(C)[C@@](CC(CC[C@]1C)O[C@]1(C(C(C1[C@]2CCCC1)=O)=O)N=O)OC)=O)OC)N=O)=O)OC2=O 0.000 description 1
Abstract
The invention relates to rapamycin or a derivative thereof which is obtained through primary preparative chromatography purification; and the preparative chromatography purification relates to normal-phase preparative chromatography purification or inverse preparative chromatography purification.
Description
Technical field
The present invention relates to the method for applying high voltage preparation system separation and purification rapamycin or derivatives thereof
Background technology
Rapamycin is the large ring triene antibiotic produced by streptomyces hygroscopicus, finds that it has in vitro and in vivo antifungal activity, particularly anti-candida albicans [people such as C.Vezina, J.Antibiot.28,721 (1975); The people such as S.N.Sehgal, J.Antibiot.28,727 (1975); The people such as H.A.Baker, J.Antibiot.31,539 (1978); United States Patent (USP) 3,929,992 and United States Patent (USP) 3,993,749].
Rapamycin has antitumor (United States Patent (USP) 4, 885, 171 and 4, 401, 653) and immunosuppressive action [FASEB3, 3411 (1989)], its purposes comprises prevention or transplantation in treating systemic erythema sore [United States Patent (USP) 5, 078, 999], pneumonia [United States Patent (USP) 5, 080, 899], insulin-dependent diabetes mellitus [inflammatory disease research association the 5th international conference (Fifth Int.Conf.Inflamm.Res.Assoc.) 121 (summary), (1990)], thickening [the Morris of the inner membrance that smooth muscle cell proliferation and injury of blood vessel cause, R.J. heart-lung transplant (Heart Lung Transplant) 11 (pt.2), 197 (1992)], adult T cell leukemia/lymthoma [EP525,960A1] and ophthalmia [EP532,862A1].Rapamycin (formula II) and rapamycin derivative comprise everolimus (formula II I), CCI-779 (structural formula IV) is always studied is used for the treatment of these and Other diseases.
Formula II
Formula II I
Structural formula IV
Everolimus has another name called 40-O-(2-hydroxyl) ethyl rapamycin, English name everolimus, and its synthesis is described in United States Patent (USP) NO.5,665,772 and International Patent Publication NO.WO94/09010 in.Its polishing purification patent is described in United States Patent (USP) NO.6,605,613B2, refines obtain crystal formation solid with ethyl acetate and n-hexane.
CCI-779 has another name called CCI-779, CCI-779, and its synthesis is described in CN1059905C and WO95/28406.Its polishing purification patent is described in the refining or methyl tertiary butyl ether(MTBE) of US2006178392A1 methyl tertiary butyl ether(MTBE) and refines with n-hexane and obtain crystal formation solid; US2007129395A1 ether is refining obtains crystal formation solid.
Rapamycin and the general purifying of derivative thereof are all adopt process for purification, and can not ensure that primary purification just can obtain high-purity sample with process for purification.WO2008065887 discloses rapamycin derivative preparative chromatography purification process, and this invention needs to prepare anti-phase preparation again by first positive just can reach purification object.Which does [method of purification about rapamycin in prior art have? how purifying rapamycin? ]
Summary of the invention
Therefore, the object of this invention is to provide a kind of method utilizing positive preparative chromatography separation and purification rapamycin or derivatives thereof;
Another goal of the invention of the present invention there is provided a kind of method utilizing reversed-phase preparative chromatography separation and purification rapamycin or derivatives thereof;
The object of the invention is to be used alone positive or reversed-phase preparative chromatography purification technique, purification high-purity rapamycin or derivatives thereof, and do not need further polishing purification, thus improve efficiency, reduce product cost.
The preparative chromatography purification process of a kind of formula (I) the rapamycin or derivatives thereof that the present invention relates to, it is characterized in that, the rapamycin or derivatives thereof of formula (I) is adsorbed in preparative chromatography Stationary liquid, obtains the product of purity more than 99% through a step chromatogram purification.
Formula (I)
Wherein R refers to hydrogen, alkyl, alkylsiloxane base, aryl alkyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl group, alkoxycarbonyl amido alkyl, acylaminoalkyl, aryl, (oxinane-2-base oxygen base) acetyl group, hydroxyacetyl, 2, 2-dimethyl-3-(oxinane-2-base oxygen base) propiono, 3-hydroxyl-2, 2-Dimethylpropanoyl, 2, 2-dimethyl [1, 3] dioxolane-4-acyl group, 2, 3-dihydroxy propiono, 3-hydroxyl-2-methylol propiono, 2, two (methylol) propiono of 2-.Wherein said alkyl or alkoxyl refer to C
1~ C
6alkyl or alkoxyl; Wherein said aryl is phenyl or substituted-phenyl, and the substituting group on phenyl is selected from C
1~ C
5alkyl, hydroxyl, nitro, sulfonic group or halogen.Wherein said formula (I) specifically refers to rapamycin, everolimus, CCI-779.
The present invention relates to preparative chromatography purification technique and be generally divided into positive preparative chromatography purifying and reversed-phase preparative chromatography purifying.The Stationary liquid that so-called positive preparative chromatography refers to is generally nonbonding silica gel or bonding, and other have polar functional group, as the bonded phase packings of amine groups and cyano group group.Because the polarity of the silicone hydroxyl of Silica Surface or other groups is comparatively strong, therefore, the order of separation is the polarity size according to each component in sample, and namely the component of polarity power is rinsed out chromatographic column at first.The mobile phase polarity Stationary liquid that compares that normal-phase chromatography uses is low, and reverse-phase chromatography filler is often based on silica gel, the Bonded Phase of the functional group that surface bond has polarity relatively weak.The mobile phase polarity that anti-phase look chromatogram uses is comparatively strong, is generally water, buffer solution and methyl alcohol, the mixtures such as own nitrile.It is that polarity combines more by force and is rushed out at first that sample flows out the order of chromatographic column, and the weak component of polarity can have stronger reservation on a column.Conventional reverse phase filler has C18, C8, C4, C6H5 etc.
In one aspect of the invention, the feature of the method for the highly purified rapamycin or derivatives thereof of the preparation that the present invention relates to is, the crude product of rapamycin or derivatives thereof positive preparative chromatography purified, obtain the product that purity is greater than 99%, wherein said positive preparative chromatography purification process refers to:
A. the sample adsorption of mobile phase will be dissolved in Stationary liquid,
B. with preparing pump, mobile phase is sucked preparative column elution samples,
C. low pole impurity is first separated, and then rapamycin or derivatives thereof product is separated, stronger polar impurity is separated, collects the stream part of closing and having rapamycin or derivatives thereof product peak, steams and desolventizes to obtain the purity sample that is greater than 99%.
In further embodiment, the Stationary liquid needed for its preparative column is nonbonding silica gel, and preferred Stationary liquid particle diameter is 3 ~ 100 μm, is more preferably 5 ~ 20 μm, more preferably 5 ~ 10 μm further, most preferably 10 μm.Experiment finds that the less then separating effect of particle diameter is better, yield is relatively high.Preferred Stationary liquid aperture is 60 ~
be more preferably 60 ~
experiment finds that the less then separating effect in aperture is better.The proterties of preferred Stationary liquid is unformed or ball-type, is more preferably ball-type, and peak type when testing the separation finding ball-type is better than unformed, and that is the separating effect of ball-type is better than unformed.
In further embodiment, described mobile phase refers to the mixed solvent of low polar organic solvent and high polar organic solvent, and the percent by volume that wherein low polar organic solvent accounts for is greater than 60%, is preferably 70% ~ 99.9%.Wherein said low polar organic solvent is selected from such as: the low polar organic solvent that alkane, halogenated alkane etc. are conventional.Preferably wherein said low polar organic solvent is selected from C
5~ C
12saturated hydrocarbons, C
1~ C
8halogenated alkane or its mixture.C
5~ C
12the preferred n-hexane of saturated hydrocarbons, normal heptane, cyclohexane, C
1~ C
8halogenated alkane be selected from carrene, chloroform; Wherein said high polar organic solvent is selected from such as: the high polar organic solvent that alcohol, ketone etc. are conventional.Preferably described high polar organic solvent is selected from C
1~ C
5alcohol, C
2~ C
10ester, C
3~ C
6ketone, C
3~ C
6cyclic ethers, acetonitrile or its mixture.C
1~ C
5alcohol be selected from methyl alcohol, ethanol, isopropyl alcohol, C
2~ C
10ester be selected from ethyl acetate, isopropyl acetate, C
3~ C
6ketone be selected from acetone, butanone, C
3~ C
6cyclic ethers be selected from oxolane, dioxane.
In the positive preparative chromatography purification process of the highly purified rapamycin or derivatives thereof of the preparation that the present invention relates to, the flow velocity of the mobile phase described in it according to the size of preparative column, prepare the uninterrupted that pump bears, and the separating degree of flowing relative impurity determines; The collection of wherein said product, because the applied sample amount of sample is larger, flat peak can be formed in collection of illustrative plates, the beginning and ending time of collection can be decided according to separating effect, if the separating effect of impurity is good especially, just out just can collect from product, if the separating effect of impurity is poor, also can collect a certain section in the middle of flat peak.Stream part containing product directly can be steamed and desolventize, obtain product after being collected.The purity HPLC of sample detects and is greater than 99%.
In another aspect of this invention, the feature of the method for the highly purified rapamycin or derivatives thereof of the preparation that the present invention relates to also is: by the crude product reversed-phase preparative chromatography purifying of rapamycin or derivatives thereof, obtain the product that purity is greater than 99%, wherein said reversed-phase preparative chromatography purification process refers to:
A. sample dissolution is adsorbed in Stationary liquid,
B. with preparing pump, mobile phase is sucked preparative column elution samples,
C. strong polar impurity is first separated, then rapamycin or derivatives thereof product is separated, low pole impurity is separated again, collects the stream part containing rapamycin or derivatives thereof product peak, with carrene extraction, concentrated, the dry sample obtaining purity and be greater than 99%.
In further embodiment, needed for its preparative column, Stationary liquid is selected from bonded silica gel, the binding groups of Stationary liquid is selected from least one in alkyl, phenyl, aIkylsilyl groups, and wherein alkyl is be selected from least one in butyl, octyl group, octadecyl, tricosyl.The particle diameter of preferred Stationary liquid is 5 ~ 100 μm, and particle diameter is more preferably 5 ~ 20 μm, more preferably 5 ~ 10 μm further, most preferably 10 μm.Experiment finds that the less then separating effect of particle diameter is better, yield is relatively high.The aperture of preferred Stationary liquid is 60 ~
more preferably 60 ~
In further embodiment, mobile phase is the aqueous solution comprising at least one organic solvent, and the organic solvent described in it is: C
1~ C
5alcohol, acetonitrile, C
3~ C
6cyclic ethers, C
1~ C
5acid.Preferably wherein said C
1~ C
5alcohol be selected from such as methyl alcohol, ethanol, isopropyl alcohol, C
3~ C
6cyclic ethers be selected from such as oxolane, dioxane, C
1~ C
5acid be selected from such as acetic acid, formic acid, trifluoroacetic acid, the percent by volume that water accounts for is 5 ~ 60%.
In the reversed-phase preparative chromatography purification process of the highly purified rapamycin or derivatives thereof of the preparation that the present invention relates to, the flow velocity of mobile phase according to the size of preparative column, prepare the uninterrupted that pump bears, and the separating degree of flowing relative impurity determines; The collection of the product described in it, because the applied sample amount of sample is larger, can form flat peak in collection of illustrative plates, if the separating effect of impurity is good especially, just out just can collect from product, if the separating effect of impurity is poor, also can collect a certain section in the middle of flat peak.After stream part containing product is collected, extract with carrene, dry, concentratedly obtain product, the purity HPLC of sample detects and is greater than 99%.
The method of the highly purified rapamycin or derivatives thereof of the preparation that the present invention relates to is further characterized in that, derivative described in it refers to everolimus, CCI-779 etc., substantially can be separated obtain highly purified rapamycin or derivatives thereof by above-mentioned preparation method.
Rapamycin forms [J.Antibiot.28,727 (1975)] by streptomyces hygroscopicus fermentation and obtains higher degree rapamycin through extracting and developing, crystallization.
Everolimus synthesis is described in United States Patent (USP) NO.5,665,772 and International Patent Publication NO.WO94/09010, take rapamycin as initiation material, react with TFMS tertiary butyl dimethyl Si base ethyl ester under DIPEA exists in carrene, through column chromatography, be hydrolyzed to obtain the higher crude product of everolimus purity.
The synthetic method of CCI-779 is described in Chinese patent CN1059905, first by 2, two (methylol) propionic acid isopropylidene ketal and 2 of 2-, the reaction of 4,6-trichloro-benzoyl chloride forms acid anhydrides and reacts with rapamycin through column chromatography again under DMAP exists, is hydrolyzed to obtain the crude product of the higher CCI-779 of purity
The present invention's advantage is compared with prior art, the crude product of the higher rapamycin or derivatives thereof of purity is obtained through synthesis, by a preparative chromatography purification process, directly can obtain highly purified rapamycin or derivatives thereof, and not need further polishing purification.The yield of the inventive method is higher, thus improves efficiency, reduces product cost.Preparative chromatography purification process of the present invention can also be amplified to technical grade very easily, without the need to carrying out large change to method, and only need according to the size of the adjustment flow velocity be in proportion of preparative column.The stability of the high purity product after the present invention's preparation is relatively good, can preserve the long period at 2 ~ 8 DEG C.
Below by embodiment, the invention will be further described; it should be understood that preparation method described in the embodiment of the present invention is only used for the present invention is described; instead of limitation of the present invention, under concept thereof of the present invention, all the scope of protection of present invention is belonged to the simple modifications of preparation method of the present invention.
Detailed description of the invention
Below by embodiment, the invention will be further described.It should be understood that preparation method described in the embodiment of the present invention is only used for the present invention is described, instead of limitation of the present invention, under concept thereof of the present invention, all the scope of protection of present invention is belonged to the simple modifications of preparation method of the present invention.
Embodiment one: the positive preparative chromatography purifying of everolimus
Instrument: preparing chromatography system.
Filler: 80g nonbonding spherical silica gel, particle diameter 10 μm, aperture
Eluant, eluent: n-hexane: ethyl acetate: methyl alcohol=80: 10: 10 (volume ratios).
Flow velocity: 20ml/min.
Determined wavelength: 278nm.
0.2g everolimus crude product (synthesized reference WO94/09010) 4ml eluant, eluent dissolves, with 5ml/min flow velocity, sample dissolution liquid being sucked pillar is adsorbed in filler, 20ml/min flow velocity wash-out, low pole impurity is first separated, and then everolimus product is separated, strong polar impurity is separated again, product peak will show flat peak preparing on collection of illustrative plates because concentration is high, collect the product of this section of flat peak, concentratedly dryly obtain everolimus, yield 70%, product purity is greater than 99%.
Embodiment two: the positive preparative chromatography purifying of everolimus
The method identical according to embodiment one changes eluant, eluent into carrene: methyl alcohol=50: 1 (volume ratio) collects that product is concentrated dry to obtain everolimus, and yield 72%, product purity is greater than 99%.
Embodiment three: the positive preparative chromatography purifying of everolimus
The method identical according to embodiment one changes eluant, eluent into n-hexane: carrene: methyl alcohol=13: 50: 1 (volume ratios) collect that product is concentrated dry to obtain everolimus, and yield 75% product purity is greater than 99%.
Embodiment four: the positive preparative chromatography purifying of everolimus
The method identical according to embodiment one changes eluant, eluent into carrene: ethyl acetate: methyl alcohol=50: 3: 1 (volume ratios) collect that product is concentrated dry to obtain everolimus, and yield 76% product purity is greater than 99%.
Embodiment five: the positive preparative chromatography purifying of everolimus
The method identical according to embodiment one changes filler into 80g particle diameter 5 μm, aperture
nonbonding spherical silica gel, with carrene: ethyl acetate: methyl alcohol=50: the eluent of 3: 1 (volume ratios), collect that product is concentrated dryly to obtain everolimus, yield 77%, product purity is greater than 99%.
Embodiment six: the positive preparative chromatography purifying of everolimus
The method identical according to embodiment one changes filler into 80g particle diameter 20 μm, aperture
positive spherical silica gel, with carrene: ethyl acetate: methyl alcohol=50: the eluent of 3: 1 (volume ratios), collect that product is concentrated dryly to obtain everolimus, yield 65% product purity is greater than 99%.
Embodiment seven: the positive preparative chromatography purifying of rapamycin
According to embodiment one, two, three, crude product everolimus replaces to rapamycin by four identical methods, the rapamycin purity prepared is greater than 99%.
Embodiment eight: the positive preparative chromatography purifying of CCI-779
According to embodiment one, two, three, crude product everolimus replaces to CCI-779 by four identical methods, the CCI-779 purity prepared is greater than 99%.
Embodiment nine: the reversed-phase preparative chromatography purifying of everolimus
Instrument: preparing chromatography system.
Filler: 350g particle diameter 10 μm, aperture
anti-phase C18 silica gel.
Eluant, eluent: methyl alcohol: oxolane: water=20: 50: 30 (volume ratios)
Flow velocity: 50ml/min.
Determined wavelength: 278nm.
0.5g everolimus crude product 10ml acetonitrile dissolves, with 10ml/min flow velocity, sample dissolution liquid being sucked pillar is adsorbed in filler, 50ml/min flow velocity wash-out, strong polar impurity is first separated, and then everolimus product is separated, low pole impurity is separated again, product peak will show flat peak preparing on collection of illustrative plates because concentration is high, collect the product of this section of flat peak, carrene extracts, dry, concentratedly dryly obtain everolimus, yield 70%, product purity is greater than 99%.
Embodiment ten: the reversed-phase preparative chromatography purifying of everolimus
The method identical according to embodiment nine changes eluant, eluent into oxolane: water=50: 50 (volume ratios) collect the extraction of product carrene, dry, concentrated doing to obtain everolimus, and yield 68%, product purity is greater than 99%.
Embodiment 11: the reversed-phase preparative chromatography purifying of everolimus
The method identical according to embodiment nine changes eluant, eluent into acetonitrile: oxolane: water=10: 35: 55 (volume ratios) collect the extraction of product carrene, dry, concentrated doing to obtain everolimus, and yield 70%, product purity is greater than 99%.
Embodiment 12: the reversed-phase preparative chromatography purifying of everolimus
The method identical according to embodiment nine changes eluant, eluent into acetonitrile: isopropyl alcohol: water=60: 5: 35 (volume ratios) collect the extraction of product carrene, dry, concentrated doing to obtain everolimus, and yield 72%, product purity is greater than 99%.
Embodiment 13: the reversed-phase preparative chromatography purifying of everolimus
The method identical according to embodiment nine changes eluant, eluent into acetonitrile: oxolane: water: acetic acid=17: 35: 48: 0.5 (volume ratios) collect the extraction of product carrene, dry, concentrated doing to obtain everolimus, yield 68%, product purity is greater than 99%.
Embodiment 14: the reversed-phase preparative chromatography purifying of everolimus
The method identical according to embodiment nine changes eluant, eluent into acetonitrile: water: acetic acid=60: 40: 0.5 (volume ratios) collect the extraction of product carrene, dry, concentrated doing to obtain everolimus, and yield 74%, product purity is greater than 99%.
Embodiment 15: the reversed-phase preparative chromatography purifying of everolimus
The method identical according to embodiment nine changes eluant, eluent into acetonitrile: water=60: 40 (volume ratios) collect the extraction of product carrene, dry, concentrated doing to obtain everolimus, and yield 75%, product purity is greater than 99%.
Embodiment 16: the reversed-phase preparative chromatography purifying of everolimus
The method identical according to embodiment nine changes eluant, eluent into acetonitrile: oxolane: water=50: 10: 40 (volume ratios) collect the extraction of product carrene, dry, concentrated doing to obtain everolimus, and yield 73%, product purity is greater than 99%.
Embodiment 17: the reversed-phase preparative chromatography purifying of everolimus
The method identical according to embodiment nine changes filler into 350g particle diameter 10 μm, aperture
anti-phase C8 silica gel, with acetonitrile: water=60: the mixed solvent wash-out of 40, collect that product is concentrated dryly to obtain everolimus, yield 70%, product purity is greater than 99%.
Embodiment 18: the reversed-phase preparative chromatography purifying of everolimus
The method identical according to embodiment nine changes filler into 250g particle diameter 20 μm, aperture
anti-phase C18 filler, with acetonitrile: water=60: the mixed solvent wash-out of 40, collect that product is concentrated dryly to obtain everolimus, yield 68%, product purity is greater than 99%.
Embodiment 19: the reversed-phase preparative chromatography purifying of everolimus
The method identical according to embodiment 11 changes filler into 250g particle diameter 20 μm, aperture
anti-phase C8 silica filler, with acetonitrile: water=60: the mixed solvent wash-out of 40, collect that product is concentrated dryly to obtain everolimus, yield 65%, product purity is greater than 99%.
The reversed-phase preparative chromatography purifying of embodiment 20 rapamycin
According to embodiment nine, ten, 11,12,13,14,15,16,17,18, crude product everolimus replaces to rapamycin by 19 identical methods, the rapamycin purity prepared is greater than 99%.
Embodiment 21: the reversed-phase preparative chromatography purifying of CCI-779
According to embodiment nine, ten, 11,12,13,14,15,16,17,18, crude product everolimus replaces to CCI-779 by 19 identical methods, the CCI-779 purity prepared is greater than 99%.
Claims (13)
1. the purification process of formula (I) rapamycin or derivatives thereof, it is characterized in that: be used alone positive preparative chromatography, the rapamycin or derivatives thereof of formula (I) is adsorbed in the Stationary liquid of described preparative chromatography, the product of purity more than 99% is obtained through a step chromatogram purification
Wherein, described formula (I) rapamycin or derivatives thereof is rapamycin, everolimus or CCI-779,
The method comprises the steps:
A. the sample adsorption of mobile phase will be dissolved in Stationary liquid,
B. with preparing pump, mobile phase is sucked preparative column elution samples,
C. rapamycin or derivatives thereof is collected,
Wherein said mobile phase refers to the mixed solvent of low polar organic solvent and high polar organic solvent, and the percent by volume that wherein low polar organic solvent accounts for is greater than 60%, and wherein said Stationary liquid is nonbonding silica gel, and particle diameter is 3 ~ 100 μm, and aperture is
shape is unformed or ball-type.
2. method according to claim 1, wherein said Stationary liquid particle diameter is 5 ~ 20 μm, and aperture is
shape is ball-type.
3. method according to claim 1, wherein said low polar organic solvent is selected from the alkane of C5 ~ C12 or cycloalkane, the halogenated alkane of C1 ~ C8 or its mixture.
4. method according to claim 3, the alkane of wherein said C5 ~ C12 or cycloalkane are selected from n-hexane, normal heptane, cyclohexane, and the halogenated alkane of C1 ~ C8 is selected from carrene, chloroform.
5. method according to claim 1, wherein said high polar organic solvent is selected from the alcohol of C1 ~ C5, the ester of C2 ~ C10, the ketone of C3 ~ C6, the cyclic ethers of C3 ~ C6, acetonitrile or its mixture.
6. method according to claim 5, the alcohol of wherein said C1 ~ C5 is selected from methyl alcohol, ethanol, isopropyl alcohol, the ester of C2 ~ C10 is selected from ethyl acetate, isopropyl acetate, and the ketone of C3 ~ C6 is selected from acetone, butanone, and the cyclic ethers of C3 ~ C6 is selected from oxolane, dioxane.
7. method according to claim 1, the percent by volume that the low polar organic solvent described in it accounts for mobile phase is 70% ~ 99.9%.
8. prepare the method for formula described in claim 1 (I) rapamycin or derivatives thereof for one kind, it is characterized in that: be used alone reversed-phase preparative chromatography, the rapamycin or derivatives thereof of formula (I) is adsorbed in the Stationary liquid of described preparative chromatography, obtain the product of purity more than 99% through a step chromatogram purification, comprise step:
A. sample dissolution is adsorbed in Stationary liquid,
B. with preparing pump, mobile phase is sucked preparative column elution samples,
C. rapamycin or derivatives thereof is collected,
Described mobile phase is the aqueous solution comprising at least one organic solvent, and the percent by volume that water accounts for is 5 ~ 60%, and wherein said Stationary liquid is selected from bonded silica gel, and particle diameter is 5 ~ 100 μm, and aperture is
Wherein said formula (I) rapamycin or derivatives thereof is everolimus or CCI-779.
9. method according to claim 8, the binding groups of wherein said Stationary liquid is selected from least one in alkyl, phenyl, aIkylsilyl groups.
10. method according to claim 9, wherein said alkyl is selected from least one in butyl, octyl group, octadecyl, tricosyl.
11. methods according to claim 8, the particle diameter of wherein said Stationary liquid is 5 ~ 20 μm, and aperture is
12. methods according to claim 8, wherein said organic solvent is: the alcohol of C1 ~ C5, acetonitrile, the cyclic ethers of C3 ~ C6, the acid of C1 ~ C5.
13. methods according to claim 12, the alcohol of wherein said C1 ~ C5 is selected from methyl alcohol, ethanol, isopropyl alcohol, and the cyclic ethers of C3 ~ C6 is selected from oxolane, dioxane, and the acid of C1 ~ C5 is selected from acetic acid, formic acid, trifluoroacetic acid.
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| CN106146536B (en) * | 2015-04-25 | 2019-07-26 | 山东新时代药业有限公司 | A kind of preparation method of everolimus |
| CN106153756B (en) * | 2015-04-25 | 2020-04-28 | 山东新时代药业有限公司 | High performance liquid chromatography for detecting rapamycin in everolimus |
| CN104892632B (en) * | 2015-06-03 | 2017-12-26 | 道中道（菏泽）制药有限公司 | A kind of everolimus of crystal form and preparation method thereof |
| CN105566348A (en) * | 2015-12-31 | 2016-05-11 | 哈药集团技术中心 | Preparation method of everolimus |
| CN109406650A (en) * | 2018-10-25 | 2019-03-01 | 美康生物科技股份有限公司 | Kit and detection method for four kinds of immunosuppressant drug concentrations in Accurate Determining people's whole blood |
| CN109851626B (en) * | 2019-01-08 | 2021-11-02 | 扬子江药业集团四川海蓉药业有限公司 | Method for separating and purifying temsirolimus |
| CN110204556B (en) * | 2019-07-16 | 2020-09-18 | 重庆医药高等专科学校 | Preparation method of (RS) -methoxy cefoxitin |
| CN110343639B (en) * | 2019-07-23 | 2021-04-27 | 中国医药集团总公司四川抗菌素工业研究所 | Streptomyces producing 15(S) -O-ethyl rapamycin |
| JP2023028473A (en) * | 2021-08-19 | 2023-03-03 | 日本マイクロバイオファーマ株式会社 | Method for producing everolimus |
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| AU2006350684B2 (en) * | 2006-11-10 | 2012-07-05 | Biocon Limited | A pure form of rapamycin and a process for recovery and purification thereof |
| JP5456321B2 (en) * | 2006-11-27 | 2014-03-26 | テルモ株式会社 | Process for producing O-alkylated rapamycin derivative and O-alkylated rapamycin derivative |
| TW200948361A (en) * | 2008-05-26 | 2009-12-01 | Chunghwa Chemical Synthesis & Biotech Co Ltd | Method for synthesizing Biolimus A9 and the like and method for improving stability of the same |
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