WO2011153874A1 - 一种3',5'-环腺苷单磷酸的结晶方法 - Google Patents
一种3',5'-环腺苷单磷酸的结晶方法 Download PDFInfo
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- WO2011153874A1 WO2011153874A1 PCT/CN2011/073121 CN2011073121W WO2011153874A1 WO 2011153874 A1 WO2011153874 A1 WO 2011153874A1 CN 2011073121 W CN2011073121 W CN 2011073121W WO 2011153874 A1 WO2011153874 A1 WO 2011153874A1
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- Prior art keywords
- acid
- solution
- monophosphate
- cyclic adenosine
- concentration
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Links
- 238000000034 method Methods 0.000 title claims abstract description 41
- 238000002425 crystallisation Methods 0.000 title claims abstract description 32
- 230000008025 crystallization Effects 0.000 title claims abstract description 27
- IVOMOUWHDPKRLL-KQYNXXCUSA-N Cyclic adenosine monophosphate Chemical compound C([C@H]1O2)OP(O)(=O)O[C@H]1[C@@H](O)[C@@H]2N1C(N=CN=C2N)=C2N=C1 IVOMOUWHDPKRLL-KQYNXXCUSA-N 0.000 title abstract description 17
- 239000002253 acid Substances 0.000 claims abstract description 40
- 239000007864 aqueous solution Substances 0.000 claims abstract description 11
- 239000012266 salt solution Substances 0.000 claims abstract description 9
- 239000000243 solution Substances 0.000 claims description 42
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 37
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 claims description 35
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 21
- 239000002126 C01EB10 - Adenosine Substances 0.000 claims description 18
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 18
- 229960005305 adenosine Drugs 0.000 claims description 18
- 238000003756 stirring Methods 0.000 claims description 14
- UDMBCSSLTHHNCD-UHFFFAOYSA-N Coenzym Q(11) Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(O)=O)C(O)C1O UDMBCSSLTHHNCD-UHFFFAOYSA-N 0.000 claims description 13
- UDMBCSSLTHHNCD-KQYNXXCUSA-N adenosine 5'-monophosphate Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)[C@H]1O UDMBCSSLTHHNCD-KQYNXXCUSA-N 0.000 claims description 12
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 10
- LNQVTSROQXJCDD-UHFFFAOYSA-N adenosine monophosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(CO)C(OP(O)(O)=O)C1O LNQVTSROQXJCDD-UHFFFAOYSA-N 0.000 claims description 9
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 7
- 229910021529 ammonia Inorganic materials 0.000 claims description 6
- 239000002585 base Substances 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 210000004907 gland Anatomy 0.000 claims description 5
- 150000004712 monophosphates Chemical class 0.000 claims description 5
- 238000003860 storage Methods 0.000 claims description 4
- 238000000967 suction filtration Methods 0.000 claims description 4
- 229910019142 PO4 Inorganic materials 0.000 claims description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 3
- 239000010452 phosphate Substances 0.000 claims description 3
- 159000000000 sodium salts Chemical class 0.000 claims description 3
- 239000003513 alkali Substances 0.000 claims description 2
- 230000000762 glandular Effects 0.000 claims description 2
- 150000003863 ammonium salts Chemical class 0.000 claims 1
- 239000012530 fluid Substances 0.000 claims 1
- 238000004321 preservation Methods 0.000 claims 1
- 238000005086 pumping Methods 0.000 claims 1
- 238000010791 quenching Methods 0.000 claims 1
- 230000000171 quenching effect Effects 0.000 claims 1
- 238000005063 solubilization Methods 0.000 claims 1
- 230000007928 solubilization Effects 0.000 claims 1
- 150000003839 salts Chemical class 0.000 abstract description 3
- 239000013078 crystal Substances 0.000 description 22
- 239000000047 product Substances 0.000 description 8
- 238000000855 fermentation Methods 0.000 description 5
- 230000004151 fermentation Effects 0.000 description 5
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- 229950006790 adenosine phosphate Drugs 0.000 description 4
- 235000011114 ammonium hydroxide Nutrition 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- -1 Cyclic adenosine phosphate salt Chemical class 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 229910052708 sodium Inorganic materials 0.000 description 3
- 238000001291 vacuum drying Methods 0.000 description 3
- 239000004254 Ammonium phosphate Substances 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 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 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- 241001494479 Pecora Species 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229910000148 ammonium phosphate Inorganic materials 0.000 description 2
- 235000019289 ammonium phosphates Nutrition 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 125000004122 cyclic group Chemical group 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- XHXFXVLFKHQFAL-UHFFFAOYSA-N phosphoryl trichloride Chemical compound ClP(Cl)(Cl)=O XHXFXVLFKHQFAL-UHFFFAOYSA-N 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 230000003068 static effect Effects 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 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 description 1
- GGZZISOUXJHYOY-UHFFFAOYSA-N 8-amino-4-hydroxynaphthalene-2-sulfonic acid Chemical compound C1=C(S(O)(=O)=O)C=C2C(N)=CC=CC2=C1O GGZZISOUXJHYOY-UHFFFAOYSA-N 0.000 description 1
- 229930024421 Adenine Natural products 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- 241000186063 Arthrobacter Species 0.000 description 1
- 241001061264 Astragalus Species 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- KJTLSVCANCCWHF-UHFFFAOYSA-N Ruthenium Chemical compound [Ru] KJTLSVCANCCWHF-UHFFFAOYSA-N 0.000 description 1
- 241000239226 Scorpiones Species 0.000 description 1
- 240000006394 Sorghum bicolor Species 0.000 description 1
- 235000011684 Sorghum saccharatum Nutrition 0.000 description 1
- 241000219094 Vitaceae Species 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000011260 aqueous acid Substances 0.000 description 1
- 235000006533 astragalus Nutrition 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 238000006065 biodegradation reaction Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 229920001429 chelating resin Polymers 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000007791 dehumidification Methods 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
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- 235000013305 food Nutrition 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 235000021021 grapes Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 210000003127 knee Anatomy 0.000 description 1
- 239000004973 liquid crystal related substance Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000010422 painting Methods 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 230000019612 pigmentation Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 229910052707 ruthenium Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 210000003802 sputum Anatomy 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 235000013547 stew Nutrition 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 210000004233 talus Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000004804 winding Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/16—Purine radicals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/16—Purine radicals
- C07H19/20—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
- C07H19/213—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids containing cyclic phosphate
Definitions
- the present invention relates to the technical field of the invention, and specifically relates to a method for crystallizing 3,5,-cycloadenosine monophosphate.
- the present invention is a new sorghum rate of 3,5 adenosine monophosphate of the knot, using the crystallization technique of ruthenium-low temperature continuation to replace the solvent precipitation method and the freeze-drying method.
- the product quality is poor and the yield is low.
- the invention provides a method for 3S , 5 «Knee scent monophosphate yung jing crystal.
- the method includes the following steps: 1) The 3,5,-cyclo-adenosine sheep phosphate ice solution and the alkali are used to produce Salt.3 5,-bad: adenosine monophosphate solution; 2) reacting 3 s , S,-cyclic adenosine phosphate solution in step 1) with acid to form 3,5'- Adenosine is a phosphoric acid; 3)
- the 0-lSd is: t0, more preferably 10 C, and the -cyclic adenosine is formed by the reaction in the step 2).
- Step 1) The 3,5,-cyclic adenosine monophosphoric acid solution is reacted with a base until the solution zen value is: ⁇ ⁇ ) ⁇ , Lou Yi is
- the base is selected from the group consisting of ammonia water and hydrogen sodium solution, and the concentration thereof is preferably 3 to :M, more preferably 3 to 5 M.
- the concentration of the 3,5," cyclic adenosine monophosphate aqueous solution in step 1) is IS-SSDg ⁇ L. preferably 15O ⁇ 20 gL o
- the image temperature in the step 2) is 10-401, preferably 15 35 ⁇ , more preferably H 35 .
- the 5,-cyclic adenosine monophosphate solution is ruminated with the acid until the solution zen value is pSl; 04.5, which is still chosen to be ⁇ 1, 5 ⁇ 3>0, more excellent For ⁇ > ⁇ 2.0 ⁇ 2.5.
- the acid is derived from one or more of acid, hydrochloric acid and phosphoric acid, and its concentration is M 1G , and it is more preferably 2-4
- the flow rate in the step 2) is preferably 0.5 to 0.5%/min, preferably 4 ⁇ ) 4%/miii, more preferably in the volume of the 3,5-cycloadenosine monophosphate solution. «0,1,4%/miii «slowly added to the 3 ⁇ :5,-cyclic adenosine monophosphate solution.
- the step of adding an acid identified fans Baht 2) further comprises a process at a stirring rate of 20 ⁇ 253 ⁇ 4 m, still preferably 90 ⁇ 2OGpm, still more preferably 1 ⁇ 150rpm stirring speed was stirred 9
- the storage time in the step 3) is 5-25 hours, preferably 5 to 2 hours, and more preferably 18 to 3 ⁇ 4 inches. ⁇ .
- the ft-edge, the ethanol washing and the vacuum drying step 3) yield the 3 ⁇ 4 engeng step.
- the method includes the following steps: 1) react with ammonia or sodium hydroxide with a concentration of 13 ⁇ 41 ⁇ 2 ⁇ ; 3,5, ⁇ a cyclic adenosine monophosphate solution to form 3,5,-cycloadenosine Phosphorus 3 ⁇ 4 «k or sodium salt solution, the solution value is pH 6.5 ⁇ 8,0; 2) The 3 5 adenine solution formed in step 1) is reacted with a concentration of 24 M acid at 3 ⁇ 4 35 Until the solution Zen Town is the product of 28 ⁇ 20 4:, obtained in step 2 > under the storage of the zen, the suction filtration, the ethanol washing strip and the true food drying, that is.
- the acid 0,;NO.tei] mto Weng Sfc in step 2) is added to the 3',5,-cyclic adenosine monophosphate solution; preferably, the process of adding the acid is captured.
- the present invention provides a process for producing 3,5,-cycloadenosine monophosphate, which comprises the step of crystallizing an aqueous solution of 3,5,-cycloadenosine monophosphate by an up-speed method.
- the number of 3,., 5'-cyclic gland monophosphate or salt solution is controlled between 6.
- the temperature is maintained at i5 ⁇ , and the stirring is controlled at 90 to 20 rpm.
- the upper acid is a citric acid, a pot acid or an acid solution.
- it is preferred that until 3, 5, the cyclic adenosine monophosphate water immersion fishing value is 3 ⁇ 4 1.5 ⁇ 3, 0, and the J 3 ⁇ 4 mixing is stopped.
- the optimal escape technique is as follows: using 3 5 ⁇ ammonia or sodium hydroxide with a concentration of 15O ⁇ 200g L 3,5, a cyclic adenosine aqueous solution of phosphoric acid reacts to produce ApH between 6.50 and ⁇ 3,5 5 Cyclic adenosine phosphate salt or Wei salt dissolved in the age, and then placed in the crystallization tank, the temperature is maintained at 3 ⁇ 35 ⁇ €, the stirring speed is controlled at 100 5 ⁇ ⁇ , according to the initial 3,, 5,-cyclic adenosine mono
- the aqueous solution of phosphoric acid (the flow rate of U ⁇ 0.43 ⁇ 4/min is added to the concentration of 2 ⁇ ship's acid until 3, 5 5," ring gland monophosphate water Yu 2,0-2.5 pB of the solution is mixed with the A static it3 ⁇ 4 stored 18-20h at 5 to 10 and then by suction filtration, wash ethanol, dry in S true, i.e., a 3-.5 ,
- the crystallization method of the 3,5 5 -cyclic adenosine monophosphate provided by the invention has a significant improvement in the yield and quality of the crystal, the crystal product, and the yield of the crystalline product is stable.
- the product purity is above 9%.
- the crystallization system and the preparation method provided by the invention have a short tube, which reduces the biodegradation of the 3,5 cyclic adenosine monophosphate and the increase of the pigmentation in the crystallization process, and the production process is carried out at normal temperature. There is no need for heating and cooling devices for special beads. Saving investment costs, the multiplication process is easier to control and repeatable.
- the present invention provides: a crystallization method, can significantly improve the quality of the final product, the crystallization yield, simple move operation was repeated sacrifice good:, proper 35, 5 '- Sun industrial production of adenosine monophosphate.
- the method for crystallizing 3' 5 5 -cycloadenosine monophosphate provided by the invention is especially suitable for the 3,5,-cyclic adenosine monophosphate synthesized by fermentation, that is, using the microbial nucleotide metabolic pathway, Astragalus is a substrate, through a series of biological enzyme hybridization reactions, the final synthesis: 3,, 5,-cyclic adenosine monophosphate, will be 3 ⁇ adenosine monoacid fermentation broth first, and the ion exchange column After the analysis, the scorpion fiber concentrates 3 ⁇ 4 benefits, and finally enters the canister for crystallization.
- the microbial synthesis method has the characteristics of low cost, small dyeing, less impurities, separation and purification process, and the like, and the crystallization method provided by the invention makes it more suitable for industrial application of the technology.
- the starting material 3,5,-cycloadenosine monodecanoic acid can be prepared by the following description of the bar to be unresolved as a limitation to the present invention, any commercially available 3,, :5 - Cyclic adenosine can be crystallized using the method provided by the present invention.
- Fermentation: Arthrobacter ⁇ 3 ⁇ )2 (deposited on January 18, 2010 in the China Microbial Culture Collection Management Committee General Microbiology Society (CGMCC), deposit address: Datun Road, Chaoyang District, Beijing, Chinese Academy of Sciences Microbiology Institute, under the accession number CGMCC No, 3584) access to seed medium (based on the weight percent of the medium, 1% glucose, 1% peptone, 0.5% yeast extract, 1% beef extract, HaCl 0.3%) The initial pH was 7.0 and cultured at 30, 240 rpm for 18 hours.
- CGMCC General Microbiology Society
- the fermentation medium in the 5L fermenter was connected to the inoculum of 10% (based on the weight percentage of the medium, 5% of the glucose, 3 ⁇ 4 ⁇ 0 4 1%, E3 ⁇ 4PQ 4 1%> MgSOschreib 1%, vegan: 0.5%, peptone; 0,5%) based on lg NaF, 3 ⁇ 4 l VB1 and 5g hypoxanthine per liter of medium, control H by TSfaOH 7.0, dissolved oxygen control is 30%, 40Grpin, underarm sputum 72 small elbow.
- the production of cyclic adenosine monophosphate is 5 Shi 0 L» Then the fermentation broth is centrifuged to remove the bacteria, and then the bacteria will be removed.
- the centrifugation supernatant ft has an ultra-molecular weight of 6000 ulton; the filter removes most of the protein, and a 3', 5,-cyclic adenosine monophosphate solution having a concentration ranging from 5 ⁇ 0 is obtained.
- the crystallization yield is the mass of 3,5-cycloadenosine monophosphate obtained by final crystallization divided by the amount of 3,5,-cycloadenosine monophosphate fed into the crystallization tank, and then Multiply by 1.00%; Purity is calculated by dividing the area of the 3,5,-cycloadenosine monophosphate by high-performance liquid chromatography to the total area of all peaks, and multiplying by 100%.
- 3M sodium hydroxide was reacted with 3,5,-cyclo-glycol monophosphoric acid aqueous solution to form a concentration of 185g/L 3', S, and the phosphate salt solution 1,53 was placed in the crystallization tank, controlled at 303 ⁇ 4 Mixing speed 200 rpm 5 to the initial 3', 5 5 - cyclic adenosine monophosphate: sodium salt solution volume of 0.4% / mto (ie 6mL mia) of the flow il ⁇ f added to a concentration of 2M sulfuric acid solution, until the pH value At 2.0, the acid is added to the static state, and the temperature is lowered to 5.
- Aqueous solution of cyclic adenosine monophosphate produces a concentration of 1 ⁇ 27gL 3% 5,-sodium adenosine monophosphate solution: 2L, control pH 8,0, placed in a crystallization tank, controlled stirring at 40 °C 3 ⁇ 43 ⁇ 44Q0rp ⁇ Initial 3 ⁇ J ⁇ adenosine monophosphate sodium salt solution volume 0.1% / mift (that is, 2ffli min r flow rate slowly added.
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
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Description
一种 3,,5,~«腺苷单磷酸的结晶方法 親求领域
本发嚇属于結 ¾技术领域, 具体涉及一种 3,,5,-环腺苷单磷酸的结晶 方法。
•i聽來
«前有关: 3 .5'-环腺苷羊#酸 晶的研究鲜有文献报道。在 2000年出 瓶鯓《最新生化药翁铺 41术》 申报道 在化学合成的 3% 5,-环腺苷单 磷酸结晶工艺, 加等体积 乙醇, 用 2mol L盐酸调 ρΗ1〜2, 即析出 白色結 , 取结磊千燥 得威品 eAMP, 母液 收, 收率 6 48%» 由 f 学会處过程中添洳了如 *啶、 三氯氧磷等毒性较大的试剂, 且反应 不完全, 劐产物多, »导致后续分离結晶收率很低。 另外, 在中国专利 ^om 中公开了直接 冷冻干燥法对 3 5,-环腺苷单磷酸浓缩液进 结 ,, 但由于冷冻千燥法杈能狳去溶液中棘水分, 虽然收率为 100%, 但产品萄純度并 Mr进一步提升, 且真空冷冻千燥设备投资大, 能源消耗 成本貌高; :限制了该技求 工业化应用。 发明内容
3,55¾ «腺苷举磷酸上:的 ¾基的解离平衡常数的 K值为 3 3〜3.9,调 节溶液萄 即可玫变它的存在形式, 如下式, 当 ρΗ ΐ:时, 3,, 5,-环腺 苷箏磷酸上的磷酸基将与碱反应生成对应的 3 5S~环腺苷单磷酸盐, 当
KpK 5 Τ, 5,-奪腺 单 酸盐将 ¾酸反应生成 3 5'-环腺苷单磷酸。
cAMP-H÷ +ΝαϋίΙ ^cAMP-NaT ^Η^Ο
cM0*-Na+ ÷HCl = ΜΡ·Η* +NaCl
:因此, 本发明钓 萄是 一种新妁高歌率的 3,, 5 环腺苷单磷酸的 结 *方 采用反瘅-低温湘续会的结晶技术取代溶剂强析方法和冷冻干燥 方法., 以克維现有 3 腺苷单磷酸结晶技术中产品质量差, 收率低的 缺, 。
本发朋的.萄¾是采 以下救术方案来实现的.。 本发明提供一种 3S,5 «膝香单磷酸翁翁晶方法., 所迷方法包括《下步骤: 1 )将 3,,5,-环腺苷羊 磷酸氷溶液与碱进行 處,生咸 .3 5,-坏:腺苷单磷酸盐溶液; 2)将步骤1) 中反禽生 鈞 3s, S,-环腺苷举磷酸盐溶液与酸进行反应,生成 3,5'- 腺苷 举磷酸; 3)于 0〜lSd 优选为: t0 , 更优选为^ 10C下保存步骤 2) 中反应生成的 -环腺苷举磷酸, 即得。
优逸地 所迷步骤 1 )申 3,5,-环腺苷单磷酸水溶液与碱进行反应, 直 至溶液禪值为: ρ ί)〜 , 楼逸为 |)H 0^9 , 更优选为 pH 6.5 .0。
所 碱选自氨水和氢氣 ¾钠溶液中的一种或凡种, 其浓度为 优选为 3~ :M, 更犹选为 3~5M。
优选地., 對迷步骤 1 ) 中的 3,,5,》环腺苷单磷酸水溶液的浓度为 IS-SSDg^L. 优选为 15O~20 gLo
优选地, 所迷歩骤 2)中的 1象温度 10-401 ,优选为 15 35Ό, 更 犹选 H 35 。
犹选地. 所¾步骤 2 )中 ,5,-环腺苷单磷酸盐溶液与酸进行反虞, 直 至溶液禪值为 pSl ;04.5, 犹选为 ρΗ1,5~3>0, 更优逸为 ι>Η 2.0^2.5。
犹选應;, 所. 酸逸自疏酸、 盐酸和磷酸中的一种或几种, 其浓度为 M 1G , 优逸为 更优选为 2-4
犹选地, 所迷步骤 2) 中的酸 按 3 5,-环腺苷单磷酸盐溶液体积的 0. ~O.5%/min的流速,优选为 4^)4%/miii, 更优选为 0,1 ,4%/miii的 «緩慢加入至 3\ :5,-环腺苷单磷酸盐溶液中。
优逸地, 辨迷步骤 2 ) 中加入酸銖过程中还包括以 20〜25¾ m的搅拌 速度, 犹选为 90~2OGpm, 更犹选为 1 〜 150rpm的搅拌速度进行搅拌9 优选地,所迷步骤 3)中 保存时间为 5-25小时,优选为 5〜2ΰ小时, 更优逸为 18~¾Ν、时。
ΈϋΑ. 所迷方 ft还 緣、 乙醇洗涤和真空千燥步骤 3 )所得产 ¾翁步驟。
衡选地,. 所迷方法 括 下步骤: 1 )用 氨水或氢氧化钠与浓 度为 1¾½Θ ; 3,,5,~环腺苷单磷酸水溶液反应, 生成 3,, 5, -环腺苷单 磷 ¾«k或钠盐溶液,溶液禪值为 pH 6.5^8,0; 2 ) 将步骤 1 )中生成的 3 5, 腺著 酸盐溶液于 ¾ 35 下与浓度为 24 M的酸进行反应,直 至溶液禪镇为禪 在 5〜 下保存步骤 2 >中得到的产物 28^20 4、:时, 抽滤、 乙醇洗条和真食干燥 , 即得。 优选地, 所迷步骤 2 ) 中的 酸 0,;NO.tei] mto翁 Sfc速緩 ft加入至 3', 5,-环腺苷单磷酸盐溶液中;优选 地, 所逮加入酸的过程中 ¾ 括 10§~150rpm的搅拌速度进 ¾拌的步 骤。
此外, 本发明 提供了一种 3,,5,-环腺苷单磷酸的制备方法, 所述方 法 括采用上速的方法使 3,, 5,-环腺苷单磷酸水溶液结晶的步骤。
本 明¾可采用 "F技暴方案来实现。 一种 35 55-环腺苷单磷酸的结 晶 法, 爾 :2 観氨恭或氲氧 ¾钠 %浓度为 15〜3.50g/L 3\ 5 环腺苷单磷 酸水溶翁 ϋ生成 疆在 之得的 3 环腺苷举磷酸铵盐或钠盐溶 波,然后置于錄晶罐申 !¾¾暴持在 ,搅拌转速控制在 20^50rpm, 按水溶 ¾#积 ;0舰 ,S%te 翁流速加入 ¾1 ^为 L01M~10.0 的酸进行 反 ¾錄晶 直到水溶液的 值为 为止, 停止搅拌, 在0~15"0下 保存 S~25 再经抽滤、 乙醇洗涤、 真空千燥即得純度高于 99%的 3,, 5'- «腺香单磷酸晶体 其中, A翁氨水或氢氧化钠浓度为 3~6M。其中, 3,., 5'-环腺香单磷酸楼数或铺盐溶液 控制在 6 之间。 其中, 优选的是温 度保持在 i5〜 ,搅拌翁逸控制在 90~20Orpm。其中 优选的是按 3,, 5,- 环腺苷举磷酸水溶 积的氣 的流速加入浓度为 0.05M^8.0M 的酸。 上逸酸为毓酸、 盆酸或 酸溶液《 其中, 优选的是直到 3,, 5,环腺 苷单磷酸水溶袭釣禪值 ¾ 1.5^3,0为止, 停 J ¾拌。 其中, 优选的是在 0~1Θ 下保存 5~紘
最优逸的歡术方案如下: 用 3 5Μ 氨水或氢氧化钠与浓度为 15O~200g L 3,,5,环腺苷举磷酸水溶液反应生 ApH为 6.5 .0之间妁 3,,55 环腺苷举磷酸餐盐或魏盐溶歲,然后置于结晶罐中,温度保持在 3 ~35·€, 搅摔转速控制在 100 5ΘΓ Μ, 按初始 3,, 5,-环腺苷单磷酸水溶液体 ·的 (U〜0.4¾/min的流速加入浓度为 2〜舰的酸, 直到 3,5 5,》环腺鲁单磷酸水
溶液的 pB值豫 2,0-2.5为 A? 静 it¾拌, 在 5~10 下保存 18-20h? 再经 抽滤、 乙醇 涤、 真 S千燥, 即得 3,, .55-环腺苷单磷酸晶体。
由此可见, 本发明所提供的 3,,55-环腺香单磷酸的结晶方法, 在结,晶 产品收率和质量方面.比常规结晶方法具有明显的提高, 结晶产品收率稳定 在 上., 产品纯度达到 9 %以上。 另外, 本发明所提供的结晶体系和 制备方 ¾操养时筒短, 减少了结晶过程中 3,, 5 环腺苷单磷酸的生物降解 和色素象翁增加, 而且生产过程在常温下进行, 无需要特珠的加热和冷却 装置., 节筠投资成本, 才乘作过程更加易于控制, 重复性好。 可见, 采用本 发明提供:的結晶方法, 可.以明显改善最终产品质量, 结晶收率高, 操作简 举, 重复牲好:, 适 35,5'~荪腺苷单磷酸的工业化生产。
本发明所提供的 3'5 5,-环腺苷单磷酸的结晶方法, 尤其适用于发酵法 合成的 3,, 5,-环腺苷单磷酸, 即利用微生物核苷酸代谢途径, 以次黄嘌呤 为底物, 通过一系列生物酶雜化反应, 最终合成: 3,, 5,-环腺苷单磷酸, 将 成 3 ~ 腺苷单 酸发酵液先.兹过颉处理和离子交换柱层析后, 然 逡翁囊纖浓縮 ¾益, 最后进入翁 罐进行结晶。 微生物合成法具有成本 低, 染牲小, 杂质少 分离純化工艺筒举等特点, 再加上本发明所提供 的结晶方法 使其更加逸合于该技术的工业化应用。 胁麵方處
根据下迷实施例, 可 更妤地理解本发明。 然而, 本领域的技术人员容 易理解, 实施例所描述的具体的物料配比、工艺条件及其结茱仅用于说明本 发明., 本应当也不会限制杈利要求书中所详细描迷的本发明。
在翁下各实施 比例中, 原料 3,, 5,-环腺苷单嶙酸可以通过以下描 述的方条制备 不處理解为对本发明的限制,任何通过商购途径获得的 3,, :5,-环腺苷举磷酸都可 ¾使用本发明提供的方法进行结晶。
1)发酵: 将节杆菌 Α3ί)2 (于 2010年 1月 18日保藏于中国微生物菌 种保藏管理委员会普通微生物會心(简称 CGMCC ),保藏单位地址: 北京 市朝阳区大屯路, 中国科学院微生物研究所, 保藏编号为 CGMCC No, 3584 )接入种子培养基(基于培养基的重量百分比计, 包舍葡萄糖 1%, 蛋白胨 1%, 酵母膏 0.5%, 牛肉膏 1%, HaCl 0.3% )中, 初始 pH为 7.0, 在 30 、 240rpm下培养 18小时。 以 10%的接种量接入 5L发酵罐中的发 酵培养基(基于培养基的重量百分比计, 包舍葡萄糖 5%, ¾ΚΡ04 1%,
E¾PQ4 1%> MgSO„ 1%, 幕素: 0.5%, 蛋白胨 ;0,5% )的基础上每升培养基 添加 lg NaF, ¾ l VB1和 5g次黄嘌呤) 中, 以 TSfaOH控制 H为 7.0, 溶氧控制为 30%, 40Grpin, Ό下发肆 72小肘。 放罐时, 环磷酸腺苷的 产量是 5 施 0 L» 然后將发酵液通过离心机.离心去除菌体,再将离心清 液 ft过截智分子量为 6000遒尔顿的超;滤膜去除大部分蛋白质, 可得到浓 度范围在 5^ 0 的 3', 5,-环腺苷单磷酸清液。
2 分离錄化; 用 5Θ¾鹏离子交换树脂 ( Amberlite IM900RF CI类) 填充霄定床, 平衡后将浓度为 5.31g/L的 3,, 5,环腺苷单磷酸清液上柱, 后吸附德和, 然后用 ,25mol L的氨水进行洗杂, 洗杂完毕后, 再 用 ¾),¾»1/L的纏 C1进行洗脫 ¾ , «液的体积为 浓度为 2.96g L, 得辑:的 3\ 5¾ -环腺苷单嶙酸純复为 95.d 收率为 96.7%。
S )粗结晶: 洗脱液再经过浓缩后 采用 3〜5M氨水或氢氧化钠与浓 度沩 1 ~2Θ%^ 3 5 坏腺苷竿 酸水溶 ¾^应, 生成 ρΗ为 6 54,0之间 5,-环腺苷举裤酸铵盐禽钠盐溶液, 然后置于结晶罐中, 温度保持在 30-35€ , 搅拌转速控制在 100〜150rpm, 按初始 3,, 5,-环腺苷单磷酸氷溶
0,l-©J¾½nin錡流速加入浓度为 2- M的酸, 直到 3,, 5¾ -环腺苷 举 酸水溶液 禪值为 2 ^3为止,停止搅拌,在 540 下保存 1^20fe, 再祭 滤、 乙醇洗涤、 真空千燥, 即得 3 5,-环腺苷单磷酸粗结晶, 其纯 度为 97^98%。
检测3 5 环腺苷单磷酸錡高效 ¾ ^色谱的最佳条件如下:
色谱柱: 汊邦 Iiehrospherf- C18色诿柱 SOmm^ mm i.<L, 5μυι); 流 动相: 甲醇- 6%体积分数)磷酸氷溶液 (用三乙胺调节 ρΗ值至 6,6) (体积比为 25:75); 流速 8mL/min; 检测波长 2S5iim; 柱温为室温; 进样体积 20 L。 釆画外标一 法定量。
在以下各实施剩中, 结晶收率是通过最终结晶得到的 3,, 5 -环腺苷单磷 酸质量除以进入结晶罐即投料的 3,, 5,-环腺苷单磷酸潘量, 再乘以 1.00%计 算得到; 純度是通过高效液相色谱^ H到的 3,, 5,-环腺苷单磷酸锋面积除以 所有峰的总面积., 再乘以 100%计算得到。 实施例 1
用 :5M氨水与 3 5*-环腺香举磷酸水溶液反应生成浓度为 lg L 3,, 5'- 环腺苷羊磷酸铵盐溶液 0.953 控制 ρΗ δ.Ο, 置于結晶罐中, 在 30°C下控制
搅 疲. iSOrpm, ¾初始 腺苷举磷酸铵 Ik溶液体积的 02%/mm (即 l.¾»Limii3 )翁 加入浓度为 2M的磷酸溶液, 直至 pH值为 2.0时停 止加:酸, 降温至 5°C, 保存 201ι, 錄晶完全后将悬浊液抽滤, 用乙醇洗涤所 得的白;色晶体, 真空干燥》 可以得到 3,,5Ί線苷单磷酸晶体 166.1g, 結晶 收率为 .5%5 純度細%。 舰铺:
n 3M氨水与 3,,5,-环腺苷 酸 反应生成就为 105g/L 3',5'- 环腺 ·单 溶液 1L, 控制 p O, 置于结晶罐中, 在 35 !下控制搅 雜速度 20¾ η 初始 3,, 5, 苷单薅酸铵盐溶^ *积的 (即 AmLfmm )的¾¾緩慢加入浓度为 3M的硫酸溶液,直至 pH值为 2.5时停止 加酸, 降湿至 5"C, 保存 1M 結晶完全后将悬法液抽滤, 用乙醇洗涤所得 嚇 晶体, 真空干燥, 可 ^得到 3,, S,-环腺苷举磷酸晶体 96.71g, 結晶收 率为 92.1:0%, 炖度 99.4%» 实翻 3
用 3M氢氧化钠与 3,, 5,-环腺香单磷酸水溶液反应生成浓度为 185g/L 3', S, 举磷酸翁盐溶液 1,53 , 置于結晶罐中, 在 30¾下控 制撤拌速度 200rpm5 以初始 3', 55-环腺苷单磷:酸钠盐溶液体积的 0.4%/mto (即 6mL mia )的流 il^f 加入浓度为 2M的硫酸溶液, 直至 pH值为 2.0 时静止加酸, 降温至 5 , 保存 结晶完全后将悬浊液抽滤, 用乙醇洗 条辨得的白色晶体, 真靈干燥, 可《得到 3,, 5,-环腺苷单磷酸晶体256.978, 结晶收率为 §2,6%, ¾ 99.3%„ 对比例 1
5Μ氨水与 3', 5,-环腺苷单磷酸水溶液反应生成浓度为 187g L 3\ 5,- 环腺苷单磷酸 溶液 §. L, 控制 pH 8, , 置于结:晶罐中, 在 30*C下控制 搅摔速度 150rpm,以初始 3', 5,-环腺苷单磷酸铵盐溶液体积的 0.2%/mk (即 lJmL min )籍流速援提加入浓度为 2M的磷酸溶液, 直至 pH值为 2.0时停 止加酸, 在 2S 下保存 结晶完全 将悬 *液抽滤, 用乙醇 涤所得的 白色晶体, 真空干燥, 可以得到 3,5 5,-环腺苷单磷酸晶体 153.8g, 结晶收率 为 8 6%, 純度 99,2¾。
翻瞻
Claims
Ϊ ,一种 腺苷举 酸 结晶方法, 其特征在于, 所迷方法 括 以下步骤:
1〉将 3,,5,-环腺苷单磷酸水溶液与碱进行反应, 生成 3,,55-环腺苷单 磷酸盐溶液;
2 将步骤 )中反应生:成 3,,5,~«腺苷单磷酸盐溶液与酸进行反应, 生處: ,55-环腺苷单磷酸;
权
3 )子 0~15。C , 优选为 0〜i C , 更优选为 S~ 0°C下保存步骤 2 )中得 到的 ,, 5, 腺苷单舞酸 即得。
2.拫据杈利要求 1所述的方法 > 其特彺在于., 所迷步骤 1 ) 中 35,55 环腺聲单峰酸水溶翁与碱进行 应, 直要至溶液 ρϊί值为 pH 6-10, 优选为 pR 6~^ r 更优选为 p 6,54.0; 优选地, 所迷碱选自氨水和氢氧化钠溶液 中潘一种或凡种,其浓度为 2~10M., 优选为 3~6铋, 更优逸为 5 M; 更 犹选 , 對述 3 5,- 腺普举磷酸氷溶液餘浓度为 15〜350 gfL, 优选为 150〜細 g L„
3..根据权利要求 1或 2所述的方法,其特征在于,所迷步骤 2 )中 3,,5, 环腺眷单 酸盐¾¾与酸进行 直至溶液 pH值为 H 1,0^5.0, 优选 ^ Β %3-4 , 更优选为 pH .2.045 优选地, 所迷酸逸自疏酸、 盐酸和 磷酸中的一讲或几种, 其浓度为 0.01^10 Μ, 犹选为 0.05^8 Μ, 更优选为
4.拫据杈利要彖1至 3中任一项所述的方法,其特征在于,所迷步骤 2 )申的反应温度为 10^4(TC , 优选为 15~3:5Ό, 更优选为 SO^SS ^
5.根据权利要求 1至 4中任一项所述的方法,其特征在于,所迷步骤 .2 ) 中的酸以按 3,, -环腺苷单磷酸盐溶液体积的 0.01-0.5 %/mk的流速, 优选为 0.04-0.4 更优选为 G,l〜0.4 %/mim的流速緩慢加入至 3 5'- 环腺苷单磷酸盐溶液中; 优选地, 所述加入酸的过程中还包括以 20^250 rpm的搅拌速度, 优选为 90〜200 rpm, 更优选为 100〜150 rpm的搅拌速度 进 掉.。
;6.根据权利要求 1至 5中任一项所述的方法,其特征在于,所迷步骤 3 )中的保存时间为 5〜25小时, 优逸为 5~20小时, 更优选为 18〜20小时。
7,根据权利要求 1至 6中任一项所述的方法 其特征在于,所迷方法
括抽 ¾、 乙醇 涤和真垒千:燥步冁 3 )所得产品的步蹶。
g.根据权 «1要求 1至 Ί中任一项所迷的方法,其特征在于,所述方法 括;《下步骤:
I ) 氨氷或氢氧化钠与浓度为 150〜2瞻 g/L的 3 55-环腺苷单 磷酸氷 液氣应,生成.3 55-环腺 举磷酸铵盐或钠盐溶液,溶液 pH值为 PH 7J-8J;
.2 )将步骤 ¾ ) 中生成的 .3,, 5,-环腺苷单磷酸盐溶液于 30 35Ό下与浓 度为 24Μ錄酸迸行反应, 直至溶液 值为 ρΗ2 ί½.5;
3 )在 下保 ·步骤 2 ) 中得到:钧产物 H20小时, 抽滤、 乙醇 洗涤和真空千譟后, 即得。
9.根据叙剁要求 S所迷的 ϋ,其特征在于, 所迷步骤 2 )中的酸以 按 3,,55- Srt苷单磷酸盐溶液体积的 Θ.Μ.4 %/βώι的流舰慢加入至 3,5 S,- IP腺苷举磷酸盐溶愛中;优逸地, 迷加入酸的过程中还包括^ 100^150 tp 的 &进行搅拌的步骤。
10.—餘 ,5,-环腺著举 酸的制备方法,其特拿在于,所迷方法包括 采用权利要求 1至 9中任一项街潘 方法使 ^,5'-环腺苷单磷酸结晶的步 骤。
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