CN1944661A - Process for preparing citicoline sodium - Google Patents
Process for preparing citicoline sodium Download PDFInfo
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- CN1944661A CN1944661A CN 200610096512 CN200610096512A CN1944661A CN 1944661 A CN1944661 A CN 1944661A CN 200610096512 CN200610096512 CN 200610096512 CN 200610096512 A CN200610096512 A CN 200610096512A CN 1944661 A CN1944661 A CN 1944661A
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- citicoline sodium
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- liquid
- yeast
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- YWAFNFGRBBBSPD-OCMLZEEQSA-M sodium;[[(2r,3s,4r,5r)-5-(4-amino-2-oxopyrimidin-1-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-oxidophosphoryl] 2-(trimethylazaniumyl)ethyl phosphate Chemical compound [Na+].O[C@@H]1[C@H](O)[C@@H](COP([O-])(=O)OP([O-])(=O)OCC[N+](C)(C)C)O[C@H]1N1C(=O)N=C(N)C=C1 YWAFNFGRBBBSPD-OCMLZEEQSA-M 0.000 title claims abstract description 45
- 229960004774 citicoline sodium Drugs 0.000 title claims abstract description 44
- 238000004519 manufacturing process Methods 0.000 title description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 36
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims abstract description 33
- 238000000926 separation method Methods 0.000 claims abstract description 26
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 25
- 238000002360 preparation method Methods 0.000 claims abstract description 23
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 15
- YHHSONZFOIEMCP-UHFFFAOYSA-O phosphocholine Chemical compound C[N+](C)(C)CCOP(O)(O)=O YHHSONZFOIEMCP-UHFFFAOYSA-O 0.000 claims abstract description 12
- 229950004354 phosphorylcholine Drugs 0.000 claims abstract description 12
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 11
- 239000008103 glucose Substances 0.000 claims abstract description 11
- 239000002904 solvent Substances 0.000 claims abstract description 10
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 9
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract description 5
- 239000000047 product Substances 0.000 claims description 38
- 239000007787 solid Substances 0.000 claims description 30
- 238000006243 chemical reaction Methods 0.000 claims description 27
- 239000000243 solution Substances 0.000 claims description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- IERHLVCPSMICTF-XVFCMESISA-N cytidine 5'-monophosphate Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(O)=O)O1 IERHLVCPSMICTF-XVFCMESISA-N 0.000 claims description 13
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 12
- 239000012043 crude product Substances 0.000 claims description 12
- 238000002425 crystallisation Methods 0.000 claims description 12
- 230000008025 crystallization Effects 0.000 claims description 12
- 239000003513 alkali Substances 0.000 claims description 11
- 230000001476 alcoholic effect Effects 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 8
- 238000001179 sorption measurement Methods 0.000 claims description 7
- 101800000263 Acidic protein Proteins 0.000 claims description 6
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 claims description 6
- 238000003916 acid precipitation Methods 0.000 claims description 6
- 230000009849 deactivation Effects 0.000 claims description 6
- 238000010612 desalination reaction Methods 0.000 claims description 6
- 238000001471 micro-filtration Methods 0.000 claims description 6
- 102000039446 nucleic acids Human genes 0.000 claims description 6
- 108020004707 nucleic acids Proteins 0.000 claims description 6
- 150000007523 nucleic acids Chemical class 0.000 claims description 6
- 239000013049 sediment Substances 0.000 claims description 6
- 238000000108 ultra-filtration Methods 0.000 claims description 6
- 238000010792 warming Methods 0.000 claims description 6
- 230000003570 biosynthesizing effect Effects 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 5
- 239000002253 acid Substances 0.000 claims description 4
- 229930183912 Cytidylic acid Natural products 0.000 claims description 3
- IERHLVCPSMICTF-UHFFFAOYSA-N cytidine monophosphate Natural products O=C1N=C(N)C=CN1C1C(O)C(O)C(COP(O)(O)=O)O1 IERHLVCPSMICTF-UHFFFAOYSA-N 0.000 claims description 3
- 230000015572 biosynthetic process Effects 0.000 claims description 2
- 239000003054 catalyst Substances 0.000 claims description 2
- 239000011259 mixed solution Substances 0.000 claims description 2
- 230000008014 freezing Effects 0.000 claims 1
- 238000007710 freezing Methods 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 7
- 102000004190 Enzymes Human genes 0.000 abstract description 6
- 108090000790 Enzymes Proteins 0.000 abstract description 6
- 230000008901 benefit Effects 0.000 abstract description 3
- 238000000605 extraction Methods 0.000 abstract description 3
- 230000008569 process Effects 0.000 abstract description 3
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 abstract description 2
- 230000036983 biotransformation Effects 0.000 abstract description 2
- 238000005516 engineering process Methods 0.000 abstract description 2
- 239000003456 ion exchange resin Substances 0.000 abstract description 2
- 229920003303 ion-exchange polymer Polymers 0.000 abstract description 2
- 239000000463 material Substances 0.000 abstract description 2
- 239000011942 biocatalyst Substances 0.000 abstract 1
- 229910052799 carbon Inorganic materials 0.000 abstract 1
- 239000000203 mixture Substances 0.000 abstract 1
- 238000000746 purification Methods 0.000 abstract 1
- NJNWCIAPVGRBHO-UHFFFAOYSA-N 2-hydroxyethyl-dimethyl-[(oxo-$l^{5}-phosphanylidyne)methyl]azanium Chemical class OCC[N+](C)(C)C#P=O NJNWCIAPVGRBHO-UHFFFAOYSA-N 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 210000002569 neuron Anatomy 0.000 description 3
- 230000006920 protein precipitation Effects 0.000 description 3
- YYROPELSRYBVMQ-UHFFFAOYSA-N 4-toluenesulfonyl chloride Chemical compound CC1=CC=C(S(Cl)(=O)=O)C=C1 YYROPELSRYBVMQ-UHFFFAOYSA-N 0.000 description 2
- ZWIADYZPOWUWEW-XVFCMESISA-N CDP Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(O)=O)O1 ZWIADYZPOWUWEW-XVFCMESISA-N 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 229960001284 citicoline Drugs 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 239000000376 reactant Substances 0.000 description 2
- PCDQPRRSZKQHHS-CCXZUQQUSA-N Cytarabine Triphosphate Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 PCDQPRRSZKQHHS-CCXZUQQUSA-N 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000000274 adsorptive effect Effects 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000005349 anion exchange Methods 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 208000029028 brain injury Diseases 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 239000007810 chemical reaction solvent Substances 0.000 description 1
- -1 choline Cytidine diphosphate ester Chemical class 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000000302 ischemic effect Effects 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Landscapes
- Saccharide Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The present invention is process of preparing citicoline sodium. The preparation process includes the biotransformation of material including 5'-cytidylate, phosphorylcholine, potassium hydroxide and glucose with yeast as the biocatalyst; extraction and separation with active carbon as the adsorbing carrier, Cl- type ion exchange resin as the separating carrier and re-compounded water-alcohol mixture as the analyzing reagent; and product purification with alcohol solvent as the crystallizing solvent. Compared with available technology, the present invention has the advantages of high product yield and high product purity.
Description
Technical field
The present invention relates to a kind of method for preparing CITICOLINE SODIUM, belong to biological pharmacy technical field.
Background technology
CITICOLINE SODIUM claims cytidine diphosphate again, is the agent of brain metabolic activation, can promote the synthetic of neuron membrane phosphide, has the reparation brain injury, and anti-hypoxia improves memory, the effect that increases intelligence, and clinical application is wider.So the preparation method of CITICOLINE SODIUM is the research topic that people relatively are concerned about.
The method for preparing at present CITICOLINE SODIUM has: 1. chemosynthesis, and utilize CMP (5 '-cytidylic acid) and phosphorylcholine as reactant, p-toluenesulfonyl chloride carries out condensation prepared as condensing agent in the presence of dinethylformamide, with 717 (Cl
-) type anion-exchange column and 711 (Cl
-) type evaporating column parting liquid alcohol precipitation prepares CITICOLINE SODIUM, its shortcoming is difficultly to separate with condensing agent, product is not suitable for medicinal; 2. enzymatic is synthetic, extracting enchylema cytidine triphosphate(CTP) (CTP) and phosphorylcholine biosynthesizing CITICOLINE SODIUM, the synthetic substrate CTP that needs of enzyme and extraction liquid of cell, synthetic price height; 3. free cell and yeast cell-free extract are synthesized CITICOLINE SODIUM, and free yeast is synthetic to need a large amount of yeast, and can only disposablely use, and has a lot of shortcomings.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of CITICOLINE SODIUM, this method is reaction carriers, selects new catalyzer and reaction solvent to carry out biosynthesizing with the enzyme in the yeast, on the basis that keeps the prior art advantage, overcome all deficiencies of prior art, make that the preparation process of CITICOLINE SODIUM is easilier carried out, product yield is higher.
Purpose of the present invention is achieved through the following technical solutions:
The preparation method of CITICOLINE SODIUM, with 5 '-cytidylic acid and phosphorylcholine as reactant, with yeast as biological catalyst biosynthesizing CITICOLINE SODIUM, it is characterized in that: may further comprise the steps---
1. with yeast, glucose, potassium hydroxide, phosphorylcholine, 5 '-cytidylic acid and water mixes back temperature adjustment to 25 ℃～40 ℃, carrying out vibration in 2～14 hours and stirring it is fully reacted;
2. reaction solution is warming up to 50 ℃～90 ℃ deactivations, carries out liquid-solid separation;
3. transferring PH is alkalescence (PH7.0～12.0), and part basic protein and nucleic acid precipitation is carried out liquid-solid separation, and then to transfer PH be acid (PH2～6), and acidic protein is precipitated, and carries out liquid-solid separation, sediment separate out;
4. use Activated Carbon Adsorption Separation, wash with pure water;
5. carry out wash-out with the molten reagent of ethanol alkali, elutriant carries out desalination, decolouring is handled, and collects liquid;
6. elutriant vacuum concentration;
7. concentrated solution adds alcoholic solvent, crystallization, liquid-solid separate crude product;
8. dissolving crude product, ultrafiltration behind the micro-filtration adds alcoholic solvent, crystallization, liquid-solid separate wet product, after the drying finished product.
The preparation method of aforesaid CITICOLINE SODIUM, step 1. in yeast be the quick-frozen yeast, freezingly form more than 5 days through being lower than subzero 20 ℃, its consumption is 30～100 kg/kg products.
The preparation method of aforesaid CITICOLINE SODIUM, the phosphorylcholine consumption of step in 1. is 3～8 kg/kg products, 5 '-the cytidylic acid consumption is 1～5 kg/kg product, the glucose consumption is 10～20 kg/kg products, the potassium hydroxide consumption is 2～5 kg/kg products, and water consumption is 800～1500 kg/kg products.
Also add MgSO in the preparation method of aforesaid CITICOLINE SODIUM, the step reaction system 1.
4Solution, its consumption are 0.5～6 kg/kg product.
The preparation method of aforesaid CITICOLINE SODIUM, step reaction system PH=4～8,65～150 rev/min stirring reaction 1. 6～10 hours was an optimum reaction condition.
The 4. middle PH of the preparation method of aforesaid CITICOLINE SODIUM, step is 2～6.
The 5. described ethanol alkali lye of the preparation method of aforesaid CITICOLINE SODIUM, step is formulated by 20% ethanol and 5% alkali lye.
The preparation method of aforesaid CITICOLINE SODIUM, step 7. and alcoholic solvent 8. be 2 times of amounts ethanol or methyl alcohol or first and second pure mixed solutions.
The outstanding substantive distinguishing features and the obvious improvement of technical solution of the present invention is mainly reflected in:
(1) the yeast reaction system is a Newtonian fuid, viscosity is little, the heterogeneous catalytic reaction system, reactive agent solution material velocity of diffusion is fast, limit some kinases and coenzyme and flow into reaction system, stop the enzyme of oxidation CITICOLINE SODIUM route of synthesis, reduce free yeast the CITICOLINE SODIUM adsorptive capacity, fixed yeast adopts the KCl solution washing can recycle more than three times, and still can keep the high enzyme activity;
(2) the synthetic CITICOLINE SODIUM of quick-frozen yeast bio only needs a small amount of yeast, has reduced expense, has improved economic benefit;
(3) adopt best reaction system to promote the synthetic of CITICOLINE SODIUM, realized the resultant quantity of peak value;
(4) product that obtains not only purity is good, and yield is higher than prior art optimum value, and actual conversion can reach more than 75%.
Description of drawings
Below in conjunction with accompanying drawing technical solution of the present invention is described further:
Fig. 1: CITICOLINE SODIUM preparation method's of the present invention main reaction formula;
Fig. 2: process flow sheet of the present invention.
Embodiment
The CITICOLINE SODIUM another name is cytidine diphosphate, citicoline, born of the same parents' two Phosphorylcholines etc., English Citicoline sodium by name, chemical name: choline Cytidine diphosphate ester list sodium salt, molecular formula: C
14H
25N
4NaO
11P
2, molecular weight: 510.31, the crystalline thing that is white in color, very easily water-soluble.CITICOLINE SODIUM is the endogenous nucleosides of natural generation in a kind of body, is the intermediate of cytolemma phosphide biosynthesizing main path, and the reparation of neuron membrane needs a large amount of CITICOLINE SODIUM; The CITICOLINE SODIUM of injection of exogenous can promote the synthetic of neuron membrane phosphide; CITICOLINE SODIUM plays an important role in keeping the various kinds of cell physiological process; In multiple because ischemic, lose blood caused cell membrane function disorder, degeneration, CITICOLINE SODIUM has tangible clinical therapeutic efficacy.
CITICOLINE SODIUM preparation method's of the present invention main reaction formula, as shown in Figure 1.The present invention is to be reaction carriers with the yeast enzyme, with 5 '-cytidylic acid, phosphorylcholine, potassium hydroxide, glucose are the mode of production that main raw material carries out bio-transformation, are absorption carrier with the activated carbon, with Cl
-Type ion exchange resin is that carrier of separating extracts and separates, with acid PH is adsorption conditions, its PH adsorbs between 2～6, with alkaline PH is elution requirement, with PH=7～12 is analysis condition, with water and the composite reagent of alcohols serves as to resolve the extraction separation that reagent carries out desorbed solution, is that recrystallisation solvent carries out the refining of finished product with the alcoholic solvent.As shown in Figure 2, the method for making of CITICOLINE SODIUM may further comprise the steps: 1. with yeast, glucose, potassium hydroxide, phosphorylcholine, 5 '-cytidylic acid and water mixes back temperature adjustment to 20 ℃～40 ℃, PH=4～8 are carried out 65～150 rev/mins of stirrings and it were fully reacted in 2～14 hours; 2. reaction solution is warming up to 50 ℃～90 ℃ deactivations, carries out liquid-solid separation; 3. transferring PH is alkalescence (PH7.0～12.0), and part basic protein and nucleic acid precipitation is carried out liquid-solid separation, and then to transfer PH be acid (PH2～6), and acidic protein is precipitated, and carries out liquid-solid separation, sediment separate out again; 4. transferring PH is 2～6, uses Activated Carbon Adsorption Separation, washs with pure water; 5. use ethanol alkali lye (20% ethanol and 5% alkali lye are formulated) reagent to carry out wash-out, elutriant carries out desalination, decolouring is handled, and collects liquid; 6. elutriant vacuum concentration; 7. concentrated solution adds 2 times of ethanol, crystallization, carry out liquid-solid separate crude product; 8. dissolving crude product, ultrafiltration behind the micro-filtration adds 2 times of ethanol and carries out crystallization, carry out again liquid-solid separate wet product, carry out dry finished product.
Wherein, step 1. yeast is freezingly below the quick-frozen yeast, warp-20 ℃ to form more than 5 days, and the quick-frozen time is long more, and effect is good more, and its consumption is 30～100 kg/kg products; The phosphorylcholine consumption is 3～8 kg/kg products, 5 '-the cytidylic acid consumption is 1～5 kg/kg product, the glucose consumption is 10～20 kg/kg products, and the potassium hydroxide consumption is 2～5 kg/kg products, and the consumption of water is 800～1500 kg/kg products; Also add MgSO in the reaction system
4Solution, its consumption are 0.5～6 kg/kg product; System also generates potassium dihydrogen phosphate; Reacting 6～10 hours is optimum reacting time.
Fig. 2 is a process flow diagram of the present invention, below in conjunction with Fig. 2 the present invention is described in further detail again.
Embodiment 1:
With 30 kilograms of quick-frozen yeast, 3 kilograms of phosphorylcholines, 1 kilogram 5 '-cytidylic acid, 10 kilograms of glucose, 2 kilograms of potassium hydroxide, 800 kg of water are mixed back temperature adjustment to 25 ℃, PH=6 carries out 65 rev/mins of stirring reactions and it was fully reacted in 6 hours; Reaction solution is warming up to 50 ℃ of deactivations, carries out liquid-solid separation; Transfer PH=8.0, part basic protein and nucleic acid precipitation are carried out liquid-solid separation, and then are transferred PH=2.5, make the acidic protein precipitation, carry out liquid-solid separation, sediment separate out; Use Activated Carbon Adsorption Separation, PH=2.5 washs with pure water; Carry out wash-out with the molten reagent of ethanol alkali, elutriant carries out desalination, decolouring is handled, and collects liquid; The elutriant vacuum concentration; Concentrated solution adds 2 times of ethanol, crystallization, liquid-solid separate crude product; Dissolving crude product, ultrafiltration behind the micro-filtration adds 2 times of ethanol, crystallization, liquid-solid separate wet product, after the drying finished product.
Embodiment 2:
With 80 kilograms of quick-frozen yeast, 4 kilograms of phosphorylcholines, 4 kilogram 5 '-cytidylic acid, 16 kilograms of glucose, 4 kilograms of potassium hydroxide, 1100 kg of water are mixed back temperature adjustment to 30 ℃, and add 0.5 kilogram of MgSO
4Solution, PH=6 carries out 120 rev/mins of stirring reactions and it was fully reacted in 8 hours; Reaction solution is warming up to 70 ℃ of deactivations, carries out liquid-solid separation; Transfer PH=10, part basic protein and nucleic acid precipitation are carried out liquid-solid separation, and then are transferred PH=4, make the acidic protein precipitation, carry out liquid-solid separation, sediment separate out; Use Activated Carbon Adsorption Separation, PH=4 washs with pure water; Carry out wash-out with the molten reagent of ethanol alkali, elutriant carries out desalination, decolouring is handled, and collects liquid; The elutriant vacuum concentration; Concentrated solution adds 2 times of methyl alcohol, crystallization, liquid-solid separate crude product; Dissolving crude product, ultrafiltration behind the micro-filtration adds 2 times of methyl alcohol, crystallization, liquid-solid separate wet product, after the drying finished product.
Embodiment 3:
With 100 kilograms of quick-frozen yeast, 8 kilograms of phosphorylcholines, 5 kilogram 5 '-cytidylic acid, 20 kilograms of glucose, 5 kilograms of potassium hydroxide, 1500 kg of water are mixed back temperature adjustment to 40 ℃, and add 6 kilograms of MgSO
4Solution, PH=8 carries out 150 rev/mins of stirring reactions and it was fully reacted in 10 hours; Reaction solution is warming up to 90 ℃ of deactivations, carries out liquid-solid separation; Transfer PH=12.0, part basic protein and nucleic acid precipitation are carried out liquid-solid separation, and then are transferred PH=5.5, make the acidic protein precipitation, carry out liquid-solid separation, sediment separate out; Use Activated Carbon Adsorption Separation, PH=5.5 washs with pure water; Carry out wash-out with the molten reagent of ethanol alkali, elutriant carries out desalination, decolouring is handled, and collects liquid; The elutriant vacuum concentration; Concentrated solution adds 2 times of first and second alcoholic solution, crystallization, liquid-solid separate crude product; Dissolving crude product, ultrafiltration behind the micro-filtration adds 2 times of first and second alcoholic solution, crystallization, liquid-solid separate wet product, after the drying finished product.
Show from above embodiment, the present invention compared with prior art, product yield and purity are all significantly better than the prior art optimum value.
More than specifically described the application example of technical solution of the present invention, they only provide as an example, are not considered as application limitations of the present invention.All operational conditions be equal to replacement, all drop within protection scope of the present invention.
Claims (8)
1. the preparation method of CITICOLINE SODIUM, with 5 '-cytidylic acid and phosphorylcholine be the main reaction thing, with yeast as biological catalyst biosynthesizing CITICOLINE SODIUM, it is characterized in that: may further comprise the steps---
1. with yeast, glucose, potassium hydroxide, phosphorylcholine, 5 '-cytidylic acid etc. and water mixes back temperature adjustment to 25 ℃～40 ℃, carrying out vibration in 2～14 hours and stirring it is fully reacted;
2. reaction solution is warming up to 50 ℃～90 ℃ deactivations, carries out liquid-solid separation;
3. transferring PH is alkalescence, PH=7.0～12.0, and part basic protein and nucleic acid precipitation is carried out liquid-solid separation, and then to transfer PH be acid, PH=2.0～6.0, and acidic protein is precipitated, and carries out liquid-solid separation, sediment separate out;
4. use Activated Carbon Adsorption Separation, wash with pure water;
5. carry out wash-out with ethanol alkali lye reagent, elutriant carries out desalination, decolouring is handled, and collects liquid;
6. elutriant vacuum concentration;
7. concentrated solution adds alcoholic solvent, crystallization, liquid-solid separate crude product;
8. dissolving crude product, ultrafiltration behind the micro-filtration adds alcoholic solvent, crystallization, liquid-solid separate wet product, after the drying finished product.
2. by the preparation method of the described CITICOLINE SODIUM of claim 1, it is characterized in that: step 1. in yeast be freezing formation more than 5 days below the quick-frozen yeast, warp-20 ℃, its consumption is 30～100 kg/kg products.
3. press the preparation method of the described CITICOLINE SODIUM of claim 1, it is characterized in that: the phosphorylcholine consumption of step in 1. is 3～8 kg/kg products, 5 '-the cytidylic acid consumption is 1～5 kg/kg product, the glucose consumption is 10～20 kg/kg products, the potassium hydroxide consumption is 2～5 kg/kg products, and water consumption is 800～1500 kg/kg products.
4. by the preparation method of the described CITICOLINE SODIUM of claim 1, it is characterized in that: also add MgSO in the step reaction system 1.
4Solution, its consumption are 0.5～6 kg/kg product.
5. by the preparation method of the described CITICOLINE SODIUM of claim 1, it is characterized in that: 1. reaction system PH=4～8 of step, carry out 65～150 rev/mins of stirring reactions and be optimum reaction condition in 6～10 hours.
6. by the preparation method of the described CITICOLINE SODIUM of claim 1, it is characterized in that: the 4. middle PH of step is 2.0～6.0.
7. by the preparation method of the described CITICOLINE SODIUM of claim 1, it is characterized in that: the 5. described ethanol alkali lye of step is that 20% ethanol and 5% alkali lye are formulated.
8. by the preparation method of the described CITICOLINE SODIUM of claim 1, it is characterized in that: step 7. and alcoholic solvent 8. be ethanol or the methyl alcohol or the first and second pure mixed solutions.
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| CN100564537C CN100564537C (en) | 2009-12-02 |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102010454B (en) * | 2010-12-02 | 2012-03-07 | 胡建荣 | Citicoline sodium compound and new method thereof |
| CN102010454A (en) * | 2010-12-02 | 2011-04-13 | 胡建荣 | Citicoline sodium compound and new method thereof |
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| CN103849666A (en) * | 2013-05-08 | 2014-06-11 | 开平牵牛生化制药有限公司 | Method for catalytically producing citicoline sodium with immobilized enzyme |
| CN103509073A (en) * | 2013-08-29 | 2014-01-15 | 洪军 | Citicoline sodium compound |
| CN103509073B (en) * | 2013-08-29 | 2016-01-06 | 洪军 | A kind of Citicoline sodium compound |
| CN103509074A (en) * | 2013-10-10 | 2014-01-15 | 江西科技师范大学 | Synthesis method of nucleoside diphosphate 6-deoxy-L-pyranose |
| CN104031105B (en) * | 2014-06-06 | 2015-07-15 | 回音必集团(江西)东亚制药有限公司 | Method for preparing citicoline sodium |
| CN104031105A (en) * | 2014-06-06 | 2014-09-10 | 浙江天冉药物研究有限公司 | Method for preparing citicoline sodium |
| CN104592335A (en) * | 2014-12-24 | 2015-05-06 | 江西科技师范大学 | Method for synthesizing nucleoside diphosphonic acid choline sodium salts |
| CN105732752A (en) * | 2016-03-18 | 2016-07-06 | 新乡学院 | Citicoline and synthetic method thereof |
| CN106146590A (en) * | 2016-06-29 | 2016-11-23 | 陈建峰 | A kind of preparation method of C14H25N4NaO11P2 |
| CN109836468A (en) * | 2017-11-24 | 2019-06-04 | 苏州华赛生物工程技术有限公司 | A method of the purifying citicoline sodium from microbial fermentation solution |
| CN113769794A (en) * | 2021-07-06 | 2021-12-10 | 沁浩膜技术(厦门)有限公司 | Ion exchange system and method for continuously removing impurities in citicoline sodium |
| CN113769794B (en) * | 2021-07-06 | 2024-04-05 | 沁浩膜技术(厦门)有限公司 | Ion exchange system and method for continuously removing impurities in citicoline sodium |
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