CN101863809B - Method for purifying doxercalciferol - Google Patents
Method for purifying doxercalciferol Download PDFInfo
- Publication number
- CN101863809B CN101863809B CN2010101977133A CN201010197713A CN101863809B CN 101863809 B CN101863809 B CN 101863809B CN 2010101977133 A CN2010101977133 A CN 2010101977133A CN 201010197713 A CN201010197713 A CN 201010197713A CN 101863809 B CN101863809 B CN 101863809B
- Authority
- CN
- China
- Prior art keywords
- doxercalciferol
- solvent
- intermediates
- crystallization
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- HKXBNHCUPKIYDM-CGMHZMFXSA-N doxercalciferol Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)/C=C/[C@H](C)C(C)C)=C\C=C1\C[C@@H](O)C[C@H](O)C1=C HKXBNHCUPKIYDM-CGMHZMFXSA-N 0.000 title claims abstract description 107
- 229960000413 doxercalciferol Drugs 0.000 title claims abstract description 106
- 238000000034 method Methods 0.000 title claims abstract description 33
- 239000002904 solvent Substances 0.000 claims abstract description 78
- 238000002425 crystallisation Methods 0.000 claims abstract description 63
- 230000008025 crystallization Effects 0.000 claims abstract description 63
- 150000002576 ketones Chemical class 0.000 claims abstract description 38
- 239000000047 product Substances 0.000 claims abstract description 36
- 238000000746 purification Methods 0.000 claims abstract description 27
- 238000004440 column chromatography Methods 0.000 claims abstract description 26
- 150000002148 esters Chemical class 0.000 claims abstract description 22
- 239000012043 crude product Substances 0.000 claims abstract description 18
- 238000000926 separation method Methods 0.000 claims abstract description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 93
- 239000000543 intermediate Substances 0.000 claims description 48
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 46
- 239000011259 mixed solution Substances 0.000 claims description 43
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 42
- ZWEHNKRNPOVVGH-UHFFFAOYSA-N 2-Butanone Chemical compound CCC(C)=O ZWEHNKRNPOVVGH-UHFFFAOYSA-N 0.000 claims description 20
- 239000000203 mixture Substances 0.000 claims description 15
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 claims description 13
- 238000010828 elution Methods 0.000 claims description 13
- 230000008569 process Effects 0.000 claims description 12
- ZAFNJMIOTHYJRJ-UHFFFAOYSA-N Diisopropyl ether Chemical compound CC(C)OC(C)C ZAFNJMIOTHYJRJ-UHFFFAOYSA-N 0.000 claims description 8
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 8
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 claims description 4
- DURPTKYDGMDSBL-UHFFFAOYSA-N 1-butoxybutane Chemical compound CCCCOCCCC DURPTKYDGMDSBL-UHFFFAOYSA-N 0.000 claims description 3
- POLCUAVZOMRGSN-UHFFFAOYSA-N dipropyl ether Chemical compound CCCOCCC POLCUAVZOMRGSN-UHFFFAOYSA-N 0.000 claims description 3
- WBJINCZRORDGAQ-UHFFFAOYSA-N formic acid ethyl ester Natural products CCOC=O WBJINCZRORDGAQ-UHFFFAOYSA-N 0.000 claims description 3
- 239000012535 impurity Substances 0.000 abstract description 24
- 239000007788 liquid Substances 0.000 abstract description 23
- 150000002170 ethers Chemical class 0.000 abstract description 4
- 239000013067 intermediate product Substances 0.000 abstract 6
- 239000000243 solution Substances 0.000 description 79
- 239000012065 filter cake Substances 0.000 description 32
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 25
- 239000003085 diluting agent Substances 0.000 description 18
- 238000004821 distillation Methods 0.000 description 18
- 238000002474 experimental method Methods 0.000 description 17
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- 238000002360 preparation method Methods 0.000 description 12
- 239000013558 reference substance Substances 0.000 description 12
- 238000012360 testing method Methods 0.000 description 12
- 239000007791 liquid phase Substances 0.000 description 10
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 9
- 238000000967 suction filtration Methods 0.000 description 9
- OFHCOWSQAMBJIW-AVJTYSNKSA-N alfacalcidol Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C\C=C1\C[C@@H](O)C[C@H](O)C1=C OFHCOWSQAMBJIW-AVJTYSNKSA-N 0.000 description 8
- 125000004432 carbon atom Chemical group C* 0.000 description 7
- 238000004587 chromatography analysis Methods 0.000 description 6
- 238000010438 heat treatment Methods 0.000 description 6
- 238000002156 mixing Methods 0.000 description 6
- 239000012071 phase Substances 0.000 description 6
- 238000001953 recrystallisation Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 238000001291 vacuum drying Methods 0.000 description 5
- 235000019166 vitamin D Nutrition 0.000 description 5
- 239000011710 vitamin D Substances 0.000 description 5
- 230000005526 G1 to G0 transition Effects 0.000 description 4
- 239000012141 concentrate Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- MECHNRXZTMCUDQ-RKHKHRCZSA-N vitamin D2 Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)/C=C/[C@H](C)C(C)C)=C\C=C1\C[C@@H](O)CCC1=C MECHNRXZTMCUDQ-RKHKHRCZSA-N 0.000 description 4
- 239000003814 drug Substances 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 238000005805 hydroxylation reaction Methods 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- 239000000741 silica gel Substances 0.000 description 3
- 229910002027 silica gel Inorganic materials 0.000 description 3
- 239000000377 silicon dioxide Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- HKXBNHCUPKIYDM-BLKIPSJVSA-N (1R,3R,5Z)-5-[(2E)-2-[(1R,3aS,7aR)-1-[(E,2R,5R)-5,6-dimethylhept-3-en-2-yl]-7a-methyl-2,3,3a,5,6,7-hexahydro-1H-inden-4-ylidene]ethylidene]-4-methylidenecyclohexane-1,3-diol Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)/C=C/[C@H](C)C(C)C)=C\C=C1\C[C@@H](O)C[C@@H](O)C1=C HKXBNHCUPKIYDM-BLKIPSJVSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 210000000988 bone and bone Anatomy 0.000 description 2
- MWKXCSMICWVRGW-UHFFFAOYSA-N calcium;phosphane Chemical compound P.[Ca] MWKXCSMICWVRGW-UHFFFAOYSA-N 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 229960001701 chloroform Drugs 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 238000010829 isocratic elution Methods 0.000 description 2
- 238000002715 modification method Methods 0.000 description 2
- 239000012454 non-polar solvent Substances 0.000 description 2
- 239000002798 polar solvent Substances 0.000 description 2
- 238000001308 synthesis method Methods 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 229940046008 vitamin d Drugs 0.000 description 2
- BPKAHTKRCLCHEA-FOPGHSPUSA-N 19-Nor-1-α,25-dihydroxyvitamin D2 Chemical compound C1([C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@@H](C=C[C@H](C)C(C)(C)O)C)=CC=C1C[C@@H](O)C[C@H](O)C1 BPKAHTKRCLCHEA-FOPGHSPUSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 101710197992 Penicillin-binding protein PbpB Proteins 0.000 description 1
- 208000005770 Secondary Hyperparathyroidism Diseases 0.000 description 1
- 229930003316 Vitamin D Natural products 0.000 description 1
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 1
- KXKVLQRXCPHEJC-UHFFFAOYSA-N acetic acid trimethyl ester Natural products COC(C)=O KXKVLQRXCPHEJC-UHFFFAOYSA-N 0.000 description 1
- 229960002535 alfacalcidol Drugs 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 230000002308 calcification Effects 0.000 description 1
- 229960005084 calcitriol Drugs 0.000 description 1
- 235000020964 calcitriol Nutrition 0.000 description 1
- 239000011612 calcitriol Substances 0.000 description 1
- GMRQFYUYWCNGIN-NKMMMXOESA-N calcitriol Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@@H](CCCC(C)(C)O)C)=C\C=C1\C[C@@H](O)C[C@H](O)C1=C GMRQFYUYWCNGIN-NKMMMXOESA-N 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000001631 haemodialysis Methods 0.000 description 1
- 230000000322 hemodialysis Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- BHEPBYXIRTUNPN-UHFFFAOYSA-N hydridophosphorus(.) (triplet) Chemical compound [PH] BHEPBYXIRTUNPN-UHFFFAOYSA-N 0.000 description 1
- 230000033444 hydroxylation Effects 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 210000001853 liver microsome Anatomy 0.000 description 1
- 239000000395 magnesium oxide Substances 0.000 description 1
- CPLXHLVBOLITMK-UHFFFAOYSA-N magnesium oxide Inorganic materials [Mg]=O CPLXHLVBOLITMK-UHFFFAOYSA-N 0.000 description 1
- AXZKOIWUVFPNLO-UHFFFAOYSA-N magnesium;oxygen(2-) Chemical compound [O-2].[Mg+2] AXZKOIWUVFPNLO-UHFFFAOYSA-N 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- TWNQGVIAIRXVLR-UHFFFAOYSA-N oxo(oxoalumanyloxy)alumane Chemical compound O=[Al]O[Al]=O TWNQGVIAIRXVLR-UHFFFAOYSA-N 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007142 ring opening reaction Methods 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 239000002594 sorbent Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940052212 zemplar Drugs 0.000 description 1
Images
Landscapes
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention provides a method for purifying doxercalciferol, which comprises the steps of: a) carrying out column chromatography separation and purification on doxercalciferol crude product, and obtaining a first intermediate product; (b) carrying out first-time crystallization on the first intermediate product in esters solvent, and obtaining a second intermediate product; (c) carrying out second-time crystallization on the second intermediate product in the mixed liquid of ethers solvent and ketones solvent, and obtaining a third intermediate product; and d) carrying out first-time crystallization on the third intermediate product in the ketones solvent, and obtaining the finished product of the doxercalciferol. Compared with the prior art, the method leads column chromatography to be carried out on the doxercalciferol crude product, then sequentially leads the crystallization to be carried out in the esters, the mixed liquid of ethers and ketones as well as the ketones for three times, and finally obtains the doxercalciferol with the purity of more than 99.80%; and the total impurity content of the doxercalciferol does not exceed 0.20%, the known impurity EPBP3 content thereof does not exceed 0.10%, and other largest unknown single impurity does not exceed 0.10%.
Description
Technical field
The present invention relates to the biological medicine technology field, relate in particular to a kind of method of purification of doxercalciferol.
Background technology
Vitamin d compounds not only can promote the absorption of bone to calcium phosphorus, guarantee the ample supply of calcium phosphorus in body, promote the normal calcification of bone, also have biological effect more widely, as suppressing cell proliferation, Cell differentiation inducing activity, adjusting body immune system etc.Research shows, vitamins D itself does not possess physiological function, only is converted into corresponding activity form, as normal physiological actions of competence exertion such as 1 alpha-hydroxy vitamin Ds.Vitamins D activates the process that becomes 1 alpha-hydroxy vitamin D and is subjected to various factors in human body, often can not normally carry out, and therefore, studies the synthetic significant of 1 alpha-hydroxy vitamin D compounds and analogue thereof.
Be used at present 1 clinical alpha-hydroxy vitamin D compounds and analogue thereof and mainly contain doxercalciferol, calcitriol, his ostelin, Zemplar and alfacalcidol etc., wherein, because doxercalciferol does not have side reaction to obtain widespread use to the hemodialysis patients with Secondary Hyperparathyroidism.
Doxercalciferol, English name are Doxercalciferol, CAS:54573-75-0, and molecular formula is C
28H
44O
2, have formula (I) structure:
Doxercalciferol enters the effect of blood by liver microsomes 25-hydroxylase, generating 1a after hydroxylation on the C-25 position, 25-OH Vintamin D2, promote the effects such as Calcium and phosphorous absorption, inducing apoptosis of tumour cell, inhibition tumor cell transfer with performance after receptors bind.
Prior art discloses the synthetic method of the 1 alpha-hydroxy vitamin D compounds such as multiple doxercalciferol and analogue thereof, as photochemical method, assemble synthesis method and structural modification method etc.Wherein, photochemical method is at first natural complex D or its analogue to be carried out 1 'alpha '-hydroxylation, then by illumination ring opening synthesis 1 alpha-hydroxy vitamin D compounds and analogue thereof; Assembling synthesis method is A to be encircled vitamins D and analogue and CD ring synthon thereof by coupling reaction, obtain 1 alpha-hydroxy vitamin D compounds and analogue thereof; The structural modification method is directly vitamins D and analogue thereof to be carried out structural modification to obtain 1 alpha-hydroxy vitamin D compounds and analogue thereof.
But, in the building-up process of doxercalciferol, other isomers impurity such as EPBP3 that can produce 1 beta-hydroxy by product or have formula (II) structure in 1 'alpha '-hydroxylation process, cause 1 alpha-hydroxy vitamin D compounds and analogue purity is low, yield is low.The doxercalciferol purity of selling in the market is generally below 99%, and single known impurities wherein can not meet requirement less than 0.15% as EPBP3, causes doxercalciferol to be difficult to directly as medicine, use.
Summary of the invention
In view of this, technical problem to be solved by this invention is to provide a kind of method of purification of doxercalciferol, and after Methods For Purification provided by the invention, more than the purity of doxercalciferol reached 99.80wt%, the content of impurity E PBP3 was no more than 0.10wt%.
The invention provides a kind of method of purification of doxercalciferol, comprising:
A) the doxercalciferol crude product is carried out column chromatography for separation, purification, obtain the first intermediates;
B) described the first intermediates are carried out crystallization for the first time in esters solvent, obtain the second intermediates;
C) described the second intermediates are carried out crystallization for the second time in the mixed solution of ether solvent and ketones solvent, obtain the 3rd intermediates;
D) described three products are carried out crystallization for the third time in ketones solvent, obtain the doxercalciferol finished product.
Preferably, described step a) in, the wash-out in the column chromatography process is gradient elution.
Preferably, moving phase while carrying out gradient elution is the mixed solution of sherwood oil and ethyl acetate, it is perhaps the mixed solution of sherwood oil and acetone, it is perhaps the mixed solution of methyl alcohol and methylene dichloride, it is perhaps the mixed solution of methyl alcohol and trichloromethane, being perhaps the mixed solution of hexanaphthene and ethyl acetate or being the mixed solution of normal hexane and ethyl acetate, is perhaps the mixed solution of isopropyl ether and ethyl acetate.
Preferably, the esters solvent described step b) is the esters solvent that contains 2～6 carbon atoms.
Preferably, described esters solvent is one or more in ethyl acetate, methyl-formiate or ethyl formate.
Preferably, the weight ratio of described esters solvent and the first intermediates is 50～70: 1.
Preferably, the ether solvent described step c) is the ether solvent that contains 2～4 carbon atoms.
Preferably, described ether solvent is one or more in ether, positive propyl ether, isopropyl ether or butyl ether.
Preferably, the weight ratio of described ether solvent and the second intermediates is 15～35: 1.
Preferably, the ketones solvent described step c) is the ketones solvent that contains 3～6 carbon atoms.
Preferably, described ketones solvent is the mixed solution of acetone, butanone or acetone and butanone.
Preferably, the weight ratio of described ketones solvent and the second intermediates is 1.5～6: 1.
Preferably, described steps d) in, the weight ratio of ketones solvent and the 3rd intermediates is 10～40: 1.
Compared with prior art, at first the present invention carries out the doxercalciferol crude product column chromatography for separation and purifies, and first intermediates that then will obtain carry out crystallization three times successively in the mixed solution of esters solvent, ethers and ketone, ketones solvent, obtain doxercalciferol.In the column chromatography process, impurity and the doxercalciferols different from doxercalciferol polarity are separated; Successively through after three crystallizatioies, separated with most of impurity of doxercalciferol different solubility in the doxercalciferol crude product, obtain purity and be the doxercalciferol more than 99.80%, its total impurities content is no more than 0.20%, the content of known impurities EPBP3 is no more than 0.10%, other maximum unknown single impurity are no more than 0.10%, can the satisfaction ostelin as the requirement of pharmaceutical applications.In addition, method of purification provided by the invention is simple to operate, and is not high to requirement for experiment condition, easy handling, and cost is lower.
Description of drawings
The high-efficient liquid phase chromatogram of the system suitability solution that Fig. 1 provides for the embodiment of the present invention 1;
Fig. 2 is the high-efficient liquid phase chromatogram of the doxercalciferol of the embodiment of the present invention 2 purifications;
Fig. 3 is the high-efficient liquid phase chromatogram of the doxercalciferol of the embodiment of the present invention 3 purifications;
Fig. 4 is the high-efficient liquid phase chromatogram of the doxercalciferol of the embodiment of the present invention 4 purifications.
Embodiment
The invention provides a kind of method of purification of doxercalciferol, comprising:
A) the doxercalciferol crude product is carried out column chromatography for separation, purification, obtain the first intermediates;
B) described the first intermediates are carried out crystallization for the first time in esters solvent, obtain the second intermediates;
C) described the second intermediates are carried out crystallization for the second time in the mixed solution of ether solvent and ketones solvent, obtain the 3rd intermediates;
D) described three products are carried out crystallization for the third time in ketones solvent, obtain the doxercalciferol finished product.
At first the present invention carries out column chromatography with the doxercalciferol crude product, and doxercalciferol crude product Semi-polarity impurity and the doxercalciferol different from doxercalciferol are separated.By column chromatography, the purity of doxercalciferol can be brought up to 91wt% from 70wt%.
Before carrying out column chromatography, need to be with the doxercalciferol dissolving crude product, in order to make the doxercalciferol dissolving crude product more abundant, the present invention preferably use ultrasonic generator with the doxercalciferol dissolving crude product in organic solvent.The present invention does not have particular restriction to described organic solvent, includes but not limited to methylene dichloride.
in carrying out the column chromatography process, the stationary phase that uses can be silica gel, also can be preferably column chromatography silica gel for sorbent materials such as aluminum oxide, magnesium oxide, gacs, and the granularity of described column chromatography silica gel is preferably 150 orders-250 order, in order to improve chromatography efficiency, preferably described doxercalciferol crude product is carried out gradient elution, while carrying out gradient elution, the moving phase of using is the mixed solution of non-polar solvent and two kinds of organic solvents of polar solvent, be preferably mixed solution or the mixed solution of sherwood oil and acetone or mixed solution or the mixed solution of methyl alcohol and trichloromethane or mixed solution or the mixed solution of normal hexane and ethyl acetate or the mixed solution of isopropyl ether and ethyl acetate of hexanaphthene and ethyl acetate of methyl alcohol and methylene dichloride of sherwood oil and ethyl acetate, the mixed solution of the mixed solution of the mixed solution of the mixed solution of sherwood oil and ethyl acetate or hexanaphthene and ethyl acetate or normal hexane and ethyl acetate or isopropyl ether and ethyl acetate more preferably, most preferably be the mixed solution of hexanaphthene and ethyl acetate, the present invention preferably carries out 4 grades of gradient elutions, while carrying out wash-out at different levels the volume ratio of preferred non-polar solvent and polar solvent be followed successively by 3: 1,2: 1,1: 1 and 1: 2.
The present invention preferably uses the elutriant in thin-layer chromatography or high performance liquid chromatography monitoring column chromatography process, concentrates after the elutriant that will contain doxercalciferol is collected, and obtains the first intermediates.In the present invention, the described concentrated vacuum-concentrcted that is preferably.
First intermediates that will obtain after column chromatography carry out crystallization for the first time in esters solvent, through after crystallization for the first time, solubleness is higher in esters solvent impurity and doxercalciferol are separated.Doxercalciferol is through column chromatography, then through esters solvent for the first time after crystallization, purity can reach 96wt%.
The effect of described esters solvent is to remove the higher impurity of solubleness as recrystallisation solvent, is preferably the esters solvent that contains 2～6 carbon atoms, more preferably one or more in ethyl acetate, methyl-formiate or ethyl formate.In order to make the more soluble and crystallization of the first intermediates, in the present invention, the weight ratio of esters solvent and the first intermediates is preferably 40～80: 1, more preferably 50～70: 1.
In order to improve the solubleness of the first intermediates, the present invention preferably is dissolved in esters solvent with the first intermediates under the condition of heating in, Heating temperature is preferably 15 ℃～50 ℃, more preferably 30 ℃～35 ℃.
Before carrying out for the first time crystallization, the present invention preferably concentrates the described ester class solution that has dissolved the first intermediates, more preferably is concentrated into 1/4～1/6 of former weight.
Crystallization is more abundant for the first time in order to make doxercalciferol, and the present invention preferably carries out crystallization for the first time under cold condition, and the low temperature during described crystallization for the first time is preferably-10 ℃～10 ℃, more preferably 0 ℃～5 ℃; The time of crystallization is preferably 1h～24h for the first time, more preferably 20h～24h.
Second intermediates that will obtain after the esters solvent crystallization carry out crystallization for the second time in the mixed solution of ether solvent and ketones solvent, in this process, with impurity and the doxercalciferol of doxercalciferol different solubility in the mixed solution of ether solvent and ketones solvent, be separated.Through after crystallization for the second time, the purity of doxercalciferol can reach 99wt%.
The mixed solution of ether solvent and ketones solvent is removed the higher impurity of solubleness as recrystallisation solvent, and wherein, described ether solvent is preferably the ether solvent that contains 2～4 carbon atoms, more preferably one or more in ether, positive propyl ether, isopropyl ether or butyl ether; Described ketones solvent is preferably the ketones solvent that contains 3～6 carbon atoms, more preferably the mixed solution of acetone, butanone or acetone and butanone.
In order to make the more soluble and crystallization of the second intermediates, in the present invention, the weight ratio of ether solvent and the second intermediates is preferably 15～35: 1, more preferably 20～30: 1; The weight ratio of ketones solvent and the second intermediates is preferably 1.5～6: 1, more preferably 3～5: 1.
In order to improve the solubleness of the second intermediates, the present invention preferably is dissolved in the mixed solution of ether solvent and ketones solvent with the second intermediates under the condition of heating in, Heating temperature is preferably 15 ℃～50 ℃, more preferably 30 ℃～35 ℃.
Before carrying out for the second time crystallization, the present invention has preferably been dissolved the ether solvent of the second intermediates and the mixed solution of ketones solvent concentrates with described, more preferably is concentrated into 1/4～1/6 of former weight.
Crystallization is more abundant for the second time in order to make doxercalciferol, and the present invention preferably carries out crystallization for the second time under cold condition, and described low temperature is preferably-10 ℃～10 ℃, more preferably 0 ℃～5 ℃; The crystallization time is preferably 1h～24h for the second time, more preferably 20h～24h.
To carry out crystallization for the third time in ketones solvent through the 3rd intermediates that obtain after crystallization for the second time, in this process, with impurity and the doxercalciferol of doxercalciferol different solubility in ketones solvent, be separated.Through after crystallization for the third time, more than the purity of doxercalciferol can reach 99.80wt%.
The effect of described ketones solvent is to remove the higher impurity of solubleness as recrystallisation solvent, is preferably the ketones solvent that contains 3～6 carbon atoms, more preferably the mixture of acetone, butanone or acetone and butanone.In order to make the more soluble and crystallization of the 3rd intermediates, in the present invention, the weight ratio of ketones solvent and the 3rd intermediates is preferably 10～40: 1, more preferably 20～30: 1.
In order to improve the solubleness of the 3rd intermediates, the present invention preferably is dissolved in ketones solvent with the 3rd intermediates under the condition of heating in, Heating temperature is preferably 15 ℃～50 ℃, more preferably 30 ℃～35 ℃.
Before carrying out for the third time crystallization, the present invention preferably concentrates the described ketone solution that has dissolved the 3rd intermediates, more preferably is concentrated into 1/4～1/6 of former weight.
Crystallization is more abundant for the third time in order to make doxercalciferol, and the present invention preferably carries out crystallization for the third time under cold condition, and described low temperature is preferably-10 ℃～10 ℃, more preferably 0 ℃～5 ℃; The crystallization time is preferably 1h～24h for the third time, more preferably 20h～24h.
The doxercalciferol that the 3rd intermediates are carried out obtaining after crystallization for the third time contains a small amount of solvent, and the present invention preferably carries out vacuum-drying with it, and obtaining can direct applied doxercalciferol finished product.Described vacuum drying temperature is preferably 20 ℃～60 ℃, more preferably 30 ℃～35 ℃.
In order to improve the crystallization rate of doxercalciferol, in above-mentioned three Crystallization Process, all can be before cooling crystallization or between add the doxercalciferol crystal as crystal seed in solution, accelerate the crystallization time, improve crystallization efficiency.
In addition, can recycle carrying out the surplus solution of crystallization after obtaining intermediates, to reduce costs.Simultaneously, because crystallization is insufficient, can contain a small amount of doxercalciferol in this surplus solution, recycling can improve the rate of recovery of doxercalciferol, recrystallisation solvent when as the surplus solution after crystallization for the first time, continuing to serve as the next batch product and carry out the first crystallization, the recrystallisation solvent when surplus solution after crystallization continues to serve as the next batch product and carries out the second crystallization for the second time, the recrystallisation solvent the when surplus solution after crystallization continues to serve as the next batch product and carries out crystallization for the third time for the third time.
Compared with prior art, at first the present invention carries out the doxercalciferol crude product column chromatography for separation and purifies, and first intermediates that then will obtain carry out crystallization three times successively in the mixed solution of esters solvent, ethers and ketone, ketones solvent, obtain doxercalciferol.In the column chromatography process, impurity and the doxercalciferols different from doxercalciferol polarity are separated; Successively through after three crystallizatioies, separated with most of impurity of doxercalciferol different solubility in the doxercalciferol crude product, obtain purity and be the doxercalciferol more than 99.80%, its total impurities content is no more than 0.20%, the content of known impurities EPBP3 is no more than 0.10%, other maximum unknown single impurity are no more than 0.10%, can the satisfaction ostelin as the requirement of pharmaceutical applications.In addition, method of purification provided by the invention is simple to operate, and is not high to requirement for experiment condition, easy handling, and cost is lower.
, in order further to understand the present invention, below in conjunction with embodiment, the method for purification of doxercalciferol provided by the invention is described in detail.
Embodiment 1
For accuracy and the circulation ratio that guarantees detected result, according to " human drugs registration technology international coordination meeting " requirement (ICH), in the present invention, each high performance liquid chromatograph that uses detects and all uses before following instrument, reagent and step to carry out the system flexibility experiment doxercalciferol:
1.1 instrument and parameter thereof
High performance liquid chromatograph: the Agilent1200 type high performance liquid chromatograph that U.S. Agilent company produces;
Detect wavelength: 205nm;
Flow velocity: 1.7mL/min;
Sample size: reference substance solution 10 μ L, system flexibility solution 5 μ L;
Column temperature: 35 ℃;
Chromatographic column: YMC-Pack ODS-AQ 150 * 4.6mm S-3 μ m;
Mobile phase composition: filter and degassed after water and filtration and degassed after the mixed solution of acetonitrile;
When detecting, described mixed solution is according to following program wash-out:
At first be the linear gradient elution that mixed solution that the water of 25: 75 and acetonitrile form carries out 15min with volume ratio, obtaining volume ratio is the water of 23: 77 and the mixed solution of acetonitrile; Then be the linear gradient elution that the mixed solution of the water of 23: 77 and acetonitrile carries out 5min with volume ratio, obtaining volume ratio is the water of 10: 90 and the mixed solution of acetonitrile; Be that the water of 10: 90 and the mixed solution of acetonitrile carry out the isocratic elution of 10min with volume ratio, then carry out the gradient elution of 6min, obtain 100% acetonitrile, finally acetonitrile is carried out the isocratic elution of 4min.
1.2 reagent
The dose volume ratio is the ethyl acetate of 1: 4 and the mixed solution of acetonitrile, and is standby as diluent;
The purity of getting 2 parts of Chongqing Taihao Pharmaceutical Co., Ltd.'s productions is 99.87% doxercalciferol 10mg, be placed in respectively the 10mL measuring bottle, add respectively diluent dissolve described doxercalciferol and be diluted to scale, shake up, obtaining respectively concentration is the 1st reference substance solution and the 2nd reference substance solution of 1.0mg/mL;
Get that 1.5mg U.S. LFP company produces, purity and be 65%, EPBP3 content is 35% doxercalciferol, be placed in the 100mL measuring bottle, add diluent dissolve described doxercalciferol and be diluted to scale, shake up, obtaining concentration is the system flexibility solution of 0.015mg/mL, and being placed in refrigerator and cooled, to freeze preservation standby.
1.3 experimental procedure
Respectively 1 pin diluent, 5 pin the 1st reference substance solution and 1 needle system adaptability solution are added in high performance liquid chromatograph, the starting time between every pin is 5min; Record the color atlas of the 1st reference substance solution to 25min, record the color atlas of diluent and system suitability solution to 50min, result is referring to Fig. 1 and table 1, table 2, the high-efficient liquid phase chromatogram of the system suitability solution that Fig. 1 provides for the embodiment of the present invention 1, wherein a is the peak of EPBP3, and b is the peak of doxercalciferol; Table 1 is the system flexibility result of the 1st reference substance solution provided by the invention, and table 2 is the resolution result of doxercalciferol provided by the invention and EPBP3.
The system flexibility result of table 1 the 1st reference substance solution provided by the invention
The resolution result of table 2 doxercalciferol provided by the invention and EPBP3
According to table 2 and Fig. 1, calculate the relative retention time of EPBP3 and the resolution of doxercalciferol and EPBP3, be respectively 0.93 and 6.78.
ICH to the system flexibility requirement of high performance chromatograph degree of detection ostelin is: the relative standard deviation of reference substance solution 5 pins is no more than 1.5%; In system suitability solution the relative retention time of EPBP3 be about 0.94 and the resolution of doxercalciferol and EPBP3 be not less than 1.5.
The relative standard deviation of the 1st reference substance solution 5 pins provided by the invention is 0.16%; In system flexibility solution the relative retention time of EPBP3 be 0.93 and the resolution of doxercalciferol and EPBP3 be 6.78, meet the system flexibility requirement, illustrate that the present invention is reliable with the detection analysis that liquid chromatograph carries out doxercalciferol.
Add respectively 2 pin the 2nd reference substance solution in high performance liquid chromatograph, starting time between every pin is 5min, record the color atlas of the 2nd reference substance solution to 25min, the color atlas that obtains and system flexibility result and the 1st reference substance solution are basic identical, illustrate that the system flexibility experiment that the present invention carries out is reliable.
30g Chongqing Taihao Pharmaceutical Co., Ltd. is produced, purity is 75.69%, the doxercalciferol dissolving crude product that contains 1.38%EPBP3 in the 90mL methylene dichloride, obtain thick solution.Thick solution is carried out column chromatography, during chromatography take 200 purpose column chromatography silica gels as stationary phase, respectively take hexanaphthene, the ethyl acetate mixture of volume ratio as 3: 1; Volume ratio is hexanaphthene, the ethyl acetate mixture of 2: 1; Volume ratio is that hexanaphthene, ethyl acetate mixture and the volume ratio of 1: 1 is that hexanaphthene, the ethyl acetate mixture of 1: 2 is that moving phase is carried out 4 grades of gradient elutions, monitor elutriant with high performance liquid chromatograph, collection contains the elutriant of doxercalciferol, then, with the concentrated evaporate to dryness of doxercalciferol, obtain 19.5g doxercalciferol evaporate to dryness product;
After carrying out system flexibility experiment as described in Example 1, get 20mg doxercalciferol evaporate to dryness product, with the diluent of preparation in embodiment 1, the evaporate to dryness product are configured to the solution of 4.0mg/mL, use high performance liquid chromatograph to test, its purity is that 92.08%, EPBP3 content is 0.31%; Wherein, solution is prepared into sample introduction and completes in 5min;
Add the 1170mL methyl-formiate in 19.48g doxercalciferol evaporate to dryness product, be heated to 38 ℃ and make solid all dissolve rear underpressure distillation, stop distillation when surplus solution weight is 225g, and with surplus solution standing crystallization 24h at the temperature of 3 ℃, then suction filtration, obtain 16.8g the first filter cake;
After carrying out system flexibility experiment as described in Example 1, get 20mg the first filter cake, with the diluent of preparation in embodiment 1, filter cake is configured to the solution of 4.0mg/mL, use high performance liquid chromatograph to test, its purity is that 97.10%, EPBP3 content is 0.09%; Wherein, solution is prepared into sample introduction and completes in 5min;
The first filter cake of 16.78g is joined in the mixing solutions of 420mL isopropyl ether and 60mL acetone, being heated to 38 ℃ makes filter cake dissolve rear underpressure distillation fully, when being 74g, surplus solution weight stops distillation, and with surplus solution standing crystallization 20h at the temperature of 3 ℃, then suction filtration, obtain 15.1g the second filter cake;
After carrying out system flexibility experiment as described in Example 1, get 20mg the second filter cake, with the diluent of preparation in embodiment 1, filter cake is configured to the solution of 4.0mg/mL, use high performance liquid chromatograph to test, its purity is that 99.14%, EPBP3 content is 0.02%; Wherein, solution is prepared into sample introduction and completes in 5min;
The second filter cake of 15.08g is joined in the mixing solutions of 380mL acetone, being heated to 38 ℃ makes filter cake dissolve rear underpressure distillation fully, when being 60g, surplus solution weight stops distillation, and with surplus solution standing crystallization 23h at the temperature of 2 ℃, then suction filtration, obtain filter cake, and filter cake is placed in vacuum drying oven, drying under reduced pressure at the temperature of 32 ℃, obtain the 12.8g finished product;
After carrying out system flexibility experiment as described in Example 1, get the 20mg finished product, finished product is configured to the solution of 4.0mg/mL with the diluent of preparation in embodiment 1, use high performance liquid chromatograph to test finished product, result is referring to Fig. 2, and Fig. 2 is the high-efficient liquid phase chromatogram of the doxercalciferol of the embodiment of the present invention 2 purifications, and Fig. 2 is analyzed, analytical results is in Table 3, and table 3 is the efficient liquid phase chromatographic analysis result of doxercalciferol of the purification of the embodiment of the present invention 2.
The efficient liquid phase chromatographic analysis result of the doxercalciferol of the purification of table 3 embodiment of the present invention 2
Wherein, each component concentration per-cent calculates according to area normalization method (deduction diluent peak), and formula is as follows:
Content %=Ai/AU*100%
Wherein, Ai is the peak area of each component;
AU is total peak area;
As shown in Table 3, after Methods For Purification provided by the invention, the purity of doxercalciferol is 99.80wt%, and EPBP3 content is 0.00%, and other single maximum unknown impuritie content are no more than 0.05wt%, and total impurities is 0.2wt%.
30g Chongqing Taihao Pharmaceutical Co., Ltd. is produced, purity is 72.58%, the doxercalciferol dissolving crude product that contains 1.43%EPBP3 in the 90mL methylene dichloride, obtain thick solution.Thick solution is carried out column chromatography, during chromatography take 200 purpose column chromatography silica gels as stationary phase, respectively take normal hexane, the ethyl acetate mixture of volume ratio as 3: 1; Volume ratio is normal hexane, the ethyl acetate mixture of 2: 1; Volume ratio is that normal hexane, ethyl acetate mixture and the volume ratio of 1: 1 is that normal hexane, the ethyl acetate mixture of 1: 2 is that moving phase is carried out 4 grades of gradient elutions, monitor elutriant with high performance liquid chromatograph, collection contains the elutriant of doxercalciferol, then, with the concentrated evaporate to dryness of doxercalciferol, obtain 18.8g doxercalciferol evaporate to dryness product;
After carrying out system flexibility experiment as described in Example 1, get 20mg doxercalciferol evaporate to dryness product, with the diluent of preparation in embodiment 1, the evaporate to dryness product are configured to the solution of 4.0mg/mL, use high performance liquid chromatograph to test, its purity is that 92.35%, EPBP3 content is 0.25%; Wherein, solution is prepared into sample introduction and completes in 5min;
Add the 1150mL methyl acetate in 18.78g evaporate to dryness product, be heated to 38 ℃ and make solid all dissolve rear underpressure distillation, stop distillation when surplus solution weight is 211g, and with surplus solution standing crystallization 20h at the temperature of 3 ℃, then suction filtration, obtain 16.2g the first filter cake;
After carrying out system flexibility experiment as described in Example 1, get 20mg the first filter cake, with the diluent of preparation in embodiment 1, the evaporate to dryness product are configured to the solution of 4.0mg/mL, use high performance liquid chromatograph to test, its purity is that 97.91%, EPBP3 content is 0.08%; Wherein, solution is prepared into sample introduction and completes in 5min;
The first filter cake of 16.18g is joined in the mixing solutions of 420mL ether and 60mL acetone, being heated to 38 ℃ makes filter cake dissolve rear underpressure distillation fully, when being 77g, surplus solution weight stops distillation, and with surplus solution standing crystallization 20h at the temperature of 3 ℃, then suction filtration, obtain 14.4g the second filter cake;
After carrying out system flexibility experiment as described in Example 1, get 20mg the second filter cake, with the diluent of preparation in embodiment 1, the evaporate to dryness product are configured to the solution of 4.0mg/mL, use high performance liquid chromatograph to test, its purity is that 99.62%, EPBP3 content is 0.01%; Wherein, solution is prepared into sample introduction and completes in 5min;
The second filter cake of 14.38g is joined in the mixing solutions of 400mL acetone, being heated to 38 ℃ makes filter cake dissolve rear underpressure distillation fully, when being 63g, surplus solution weight stops distillation, and with surplus solution standing crystallization 20h at the temperature of 2 ℃, then suction filtration, obtain filter cake, and filter cake is placed in vacuum drying oven, drying under reduced pressure at the temperature of 32 ℃, obtain the 12.1g finished product;
After carrying out system flexibility experiment as described in Example 1, get the 20mg finished product, the evaporate to dryness product are configured to the solution of 4.0mg/mL with the diluent of preparation in embodiment 1, use high performance liquid chromatograph to test, result is referring to Fig. 3, and Fig. 3 is the high-efficient liquid phase chromatogram of the doxercalciferol of the embodiment of the present invention 3 purifications; As shown in Figure 3, the purity of doxercalciferol is 99.80wt%, and EPBP3 content is 0.00%, and other single maximum unknown impuritie content are no more than 0.05wt%, and total impurities is no more than 0.2wt%.
30g Chongqing Taihao Pharmaceutical Co., Ltd. is produced, purity is 74.33%, the doxercalciferol dissolving crude product that contains 1.27%EPBP3 in the 90mL methylene dichloride, obtain thick solution.Thick solution is carried out column chromatography, during chromatography take 200 purpose column chromatography silica gels as stationary phase, respectively take hexanaphthene, the ethyl acetate mixture of volume ratio as 3: 1; Volume ratio is hexanaphthene, the ethyl acetate mixture of 2: 1; Volume ratio is that hexanaphthene, ethyl acetate mixture and the volume ratio of 1: 1 is that hexanaphthene, the ethyl acetate mixture of 1: 2 is that moving phase is carried out 4 grades of gradient elutions, monitor elutriant with high performance liquid chromatograph, collection contains the elutriant of doxercalciferol, then, with the concentrated evaporate to dryness of doxercalciferol, obtain 19.2g doxercalciferol evaporate to dryness product;
After carrying out system flexibility experiment as described in Example 1, get 20mg doxercalciferol evaporate to dryness product, with the diluent of preparation in embodiment 1, the evaporate to dryness product are configured to the solution of 4.0mg/mL, use high performance liquid chromatograph to test, its purity is that 92.0%, EPBP3 content is 0.26%; Wherein, solution is prepared into sample introduction and completes in 5min;
Add the 1200mL methyl-formiate in 19.18g evaporate to dryness product, be heated to 38 ℃ and make solid all dissolve rear underpressure distillation, stop distillation when surplus solution weight is 207g, and with surplus solution standing crystallization 20h at the temperature of 3 ℃, then suction filtration, obtain 16.9g the first filter cake;
After carrying out system flexibility experiment as described in Example 1, get 20mg the first filter cake, with the diluent of preparation in embodiment 1, the evaporate to dryness product are configured to the solution of 4.0mg/mL, use high performance liquid chromatograph to test, its purity is that 97.77%, EPBP3 content is 0.09%; Wherein, solution is prepared into sample introduction and completes in 5min;
The first filter cake of 16.88g is joined in the mixing solutions of 420mL isopropyl ether and 60mL acetone, being heated to 38 ℃ makes filter cake dissolve rear underpressure distillation fully, when being 66g, surplus solution weight stops distillation, and with surplus solution standing crystallization 20h at the temperature of 3 ℃, then suction filtration, obtain 15.1g the second filter cake;
After carrying out system flexibility experiment as described in Example 1, get 20mg the second filter cake, with the diluent of preparation in embodiment 1, the evaporate to dryness product are configured to the solution of 4.0mg/mL, use high performance liquid chromatograph to test, its purity is that 99.56%, EPBP3 content is 0.03%; Wherein, solution is prepared into sample introduction and completes in 5min;
The second filter cake of 15.08g is joined in the mixing solutions of 400mL acetone, being heated to 38 ℃ makes filter cake dissolve rear underpressure distillation fully, when being 59g, surplus solution weight stops distillation, and with surplus solution standing crystallization 20h at the temperature of 2 ℃, then suction filtration, obtain filter cake, and filter cake is placed in vacuum drying oven, drying under reduced pressure at the temperature of 32 ℃, obtain the 12.9g finished product;
After carrying out system flexibility experiment as described in Example 1, get the 20mg finished product, the evaporate to dryness product are configured to the solution of 4.0mg/mL with the diluent of preparation in embodiment 1, use high performance liquid chromatograph to test, result is referring to Fig. 4, and Fig. 4 is the high-efficient liquid phase chromatogram of the doxercalciferol of the embodiment of the present invention 4 purifications; As shown in Figure 4, the purity of doxercalciferol is 99.82wt%, and EPBP3 content is 0.02wt%, and other single maximum unknown impuritie content are no more than 0.05wt%, and total impurities is no more than 0.2wt%.
The explanation of above embodiment just is used for helping to understand method of the present invention and core concept thereof.Should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention, can also carry out some improvement and modification to the present invention, these improvement and modification also fall in the protection domain of the claims in the present invention.
Claims (5)
1. the method for purification of a doxercalciferol, is characterized in that, comprising:
A) the doxercalciferol crude product is carried out column chromatography for separation, purification, obtain the first intermediates; Wash-out in the column chromatography process is gradient elution; Moving phase while carrying out gradient elution is the mixed solution of hexanaphthene and ethyl acetate, is perhaps the mixed solution of normal hexane and ethyl acetate;
B) described the first intermediates are carried out crystallization for the first time in esters solvent, obtain the second intermediates; Described esters solvent is one or more in ethyl acetate, methyl-formiate or ethyl formate;
C) described the second intermediates are carried out crystallization for the second time in the mixed solution of ether solvent and ketones solvent, obtain the 3rd intermediates; Described ether solvent is one or more in ether, positive propyl ether, isopropyl ether or butyl ether; Described ketones solvent is the mixed solution of acetone, butanone or acetone and butanone;
D) described the 3rd intermediates are carried out crystallization for the third time in ketones solvent, obtain the doxercalciferol finished product, described ketones solvent is the mixture of acetone, butanone or acetone and butanone.
2. method of purification according to claim 1, is characterized in that, in described step b), the weight ratio of described esters solvent and the first intermediates is 50 ~ 70:1.
3. method of purification according to claim 1, is characterized in that, in described step c), the weight ratio of described ether solvent and the second intermediates is 15 ~ 35:1.
4. method of purification according to claim 1, is characterized in that, in described step c), the weight ratio of described ketones solvent and the second intermediates is 1.5 ~ 6:1.
5. method of purification according to claim 1, is characterized in that, in described step d), the weight ratio of ketones solvent and the 3rd intermediates is 10 ~ 40:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101977133A CN101863809B (en) | 2010-05-12 | 2010-06-11 | Method for purifying doxercalciferol |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010176438.7 | 2010-05-12 | ||
| CN201010176438 | 2010-05-12 | ||
| CN2010101977133A CN101863809B (en) | 2010-05-12 | 2010-06-11 | Method for purifying doxercalciferol |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101863809A CN101863809A (en) | 2010-10-20 |
| CN101863809B true CN101863809B (en) | 2013-11-13 |
Family
ID=42955773
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010101977133A Expired - Fee Related CN101863809B (en) | 2010-05-12 | 2010-06-11 | Method for purifying doxercalciferol |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101863809B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2698160C (en) * | 2010-03-30 | 2017-07-25 | Alphora Research Inc. | Stabilized doxercalciferol and process for manufacturing the same |
| CN105254548A (en) * | 2015-10-29 | 2016-01-20 | 无锡福祈制药有限公司 | Doxercalciferol purification method |
| CN105237452B (en) * | 2015-11-01 | 2018-02-13 | 南京海融制药有限公司 | A kind of new crystalline form of doxercalciferol and preparation method thereof |
| CN106770849B (en) * | 2016-11-29 | 2018-08-14 | 无锡福祈制药有限公司 | A kind of detection method measuring doxercalciferol and its impurities |
| CN106857403A (en) * | 2017-03-22 | 2017-06-20 | 广东海洋大学 | 25‑OH‑D3Promote the application of 1 21 age in days white meat-type chickens Tibia Developments |
| CN107064359B (en) * | 2017-04-17 | 2019-04-23 | 南京健友生化制药股份有限公司 | The calculation method of content in doxercalciferol injection |
| CN116023313A (en) * | 2022-12-27 | 2023-04-28 | 南京海鲸药业股份有限公司 | Docalcitol compound crystal and capsule thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1709869A (en) * | 2005-05-31 | 2005-12-21 | 台州市海盛化工有限公司 | Vitamin D separating, purifying and crystallizing method |
-
2010
- 2010-06-11 CN CN2010101977133A patent/CN101863809B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1709869A (en) * | 2005-05-31 | 2005-12-21 | 台州市海盛化工有限公司 | Vitamin D separating, purifying and crystallizing method |
Non-Patent Citations (1)
| Title |
|---|
| 孙博等.度骨化醇的合成.《中国新药杂志》.2009,第18卷(第13期),1243,1244,1276. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101863809A (en) | 2010-10-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101863809B (en) | Method for purifying doxercalciferol | |
| CN112748206B (en) | High performance liquid chromatography analysis method simultaneously suitable for determination of 24 antiepileptic drugs in human plasma | |
| Zheng et al. | Composition and serum antioxidation of the main flavonoids from fermented vine tea (Ampelopsis grossedentata) | |
| CN102584918B (en) | Method for preparing high-purity baicalin | |
| CN108802255B (en) | Method for determining content of ganoderma lucidum triterpene component in compound preparation | |
| Płonka et al. | Multivitamin analysis of fruits, fruit–vegetable juices, and diet supplements | |
| CN104730171B (en) | Radix Paeoniae Rubra, Flos Lonicerae multi-target ingredient content assaying method in a kind of compound Chinese medicinal preparation | |
| CN103623433A (en) | Modified zebra fish drug metabolism modeling method | |
| CN105296560A (en) | Preparation method of phosphatidylserine | |
| CN101735030A (en) | Method for preparing 6-shogaol | |
| Dang et al. | Large-scale preparative isolation of bergenin standard substance from Saxifraga atrata using polyamide coupled with MCI GEL® CHP20P as stationary phases in medium pressure chromatography | |
| CN103494843B (en) | Yellow mushroom standardized component manufacturing method and application of components to treatment of lung cancer | |
| Roberts et al. | Assay of retinoids in biological samples by reverse-phase high-pressure liquid chromatography | |
| CN107727754A (en) | A kind of HPLC fingerprint atlas detection methods of Xiaojing | |
| Silva Jr et al. | Metabolism of retinoic acid in vivo in the vitamin A-deficient rat | |
| RU2647451C1 (en) | Method of the vitamin k1 content determining in products of plant origin | |
| CN101658598B (en) | Method for extracting and enriching alisma total triterpenic ketone alcohol components from alisma | |
| CN108152431A (en) | A kind of construction method and quality determining method of rhodiola root broken wall medicine materical crude slice HPLC finger-prints | |
| CN102539220B (en) | Analysis method for metabolin of A-type proanthocyanidins | |
| CN109251161B (en) | Preparation method of 2-tryptophan bisulfite | |
| CN102670670B (en) | Preparation method of ginkgo dipyridolum injection with high content of ginkgo terpene lactones | |
| Shimizu et al. | Separation and quantitative analysis of saikosaponins by high-performance liquid chromatography | |
| CN109406685A (en) | A kind of efficient liquid-phase chromatography method separating Carfilzomib and its isomers | |
| CN103235067A (en) | Method for enriching gallotannin with antioxidation activity from mangos | |
| CN108663442A (en) | A kind of inspection method of the Alfacalcidol tablet in relation to substance |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| ASS | Succession or assignment of patent right |
Owner name: UNIV ZUERICH AZAD PHARMA AG Effective date: 20121220 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20121220 Address after: 400039 Chongqing Jiulongpo Erlang venture Road No. 105 C-3 Applicant after: Chongqing Taihao Pharmaceutical Co., Ltd. Applicant after: Univ Zuerich Applicant after: AZAD Pharmaceutical Co., Ltd. Address before: 400039 Chongqing Jiulongpo Erlang venture Road No. 105 C-3 Applicant before: Chongqing Taihao Pharmaceutical Co., Ltd. |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C53 | Correction of patent for invention or patent application | ||
| CB03 | Change of inventor or designer information |
Inventor after: Li Jing Inventor after: Hans Ulrich Beek Purcell Inventor after: Budde Thomas Inventor after: Chen Bo Inventor after: Yang Xianmei Inventor before: Li Jing Inventor before: Chen Bo Inventor before: Ying Zhenpei Inventor before: Yang Xianmei |
|
| COR | Change of bibliographic data |
Free format text: CORRECT: INVENTOR; FROM: LI JING CHEN BO YING ZHENPEI YANG XIANMEI TO: LI JING HANS ULRICH BICZAEL THOMAS BADE CHEN BO YANG XIANMEI |
|
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20131113 Termination date: 20200611 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |