CN104262438A - Method for synthesizing symmetric bi-ribonucleoside triphosphate sodium salt, bi-ribonucleoside tetraphosphate sodium salt and bi-ribonucleoside pentaphosphates odium salt - Google Patents
Method for synthesizing symmetric bi-ribonucleoside triphosphate sodium salt, bi-ribonucleoside tetraphosphate sodium salt and bi-ribonucleoside pentaphosphates odium salt Download PDFInfo
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- CN104262438A CN104262438A CN201410404196.0A CN201410404196A CN104262438A CN 104262438 A CN104262438 A CN 104262438A CN 201410404196 A CN201410404196 A CN 201410404196A CN 104262438 A CN104262438 A CN 104262438A
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- ribonucleoside
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Abstract
The invention belongs to the field of chemical synthesis, and relates to a method for synthesizing symmetric bi-ribonucleoside triphosphate sodium salt, bi-ribonucleoside tetraphosphate sodium salt and bi-ribonucleoside pentaphosphates sodium salt. By using 4,5-dicyanoimidazole as an activating reagent, monophosphate, pyrophosphate and triphosphate serving as nucleophilic reagents are coupled with more than twice of excessive nucleoside phosphoryl piperidine triethylamine salts to produce corresponding nucleoside polyphosphate intermediates, and the reaction of the nucleoside polyphosphoric acid intermediates with the remaining nucleoside phosphoryl piperidine is continually performed in one-pot reaction to produce corresponding symmetric bi-ribonucleoside polyphosphate sodium salt. The method is simple, and the separation yield of products is high.
Description
Technical field
The invention belongs to the chemical preparation techniques field of natural product class biochemical reagents, relate to new chemical synthetic method and the technique of a series of double-core guanosine triphosphate, four phosphoric acid and five sodium phosphate salts.
Technical background
Symmetric double nucleoside polyphosphates (symmetrical dinucleoside polyphosphates, Np
n ns) be the special biomolecules that a class is extensively present in body.This kind of material plays important effect in the Activities of life system.Such as, two adenosine five phosphoric acid (Ap
5a) good restraining effect is had to the platelet aggregation of being lured by ADP-, and two guanosine triphosphate (Gp
3g) good promoter action is served to the regeneration of vascular smooth muscle cell.Compared with single nucleoside polyphosphates, two nucleoside polyphosphates in vivo metabolism has the longer transformation period, and this compounds also has significant application value as the inhibitor of P2Y acceptor in medicine and pharmacology.Ap
4a was once tested in clinical operation the blood pressure controlling patient, and two uridine four phosphoric acid (Up
4u) be used for treating dry eye syndrome as a kind of medicine.
At present, the symmetric double nucleoside polyphosphates synthetic method of bibliographical information is very limited, most importantly with Nucleotide monophosphates or nucleoside diphosphate for raw material, obtain target product by condensation reagent dicyclohexylcarbodiimide (DCC), carbonyl dimidazoles (CDI) or sulfonymethyl imidazole salts and nucleoside diphosphate or ribonucleoside triphosphote condensation.But, above method nucleoside diphosphate and ribonucleoside triphosphote etc. all will be used expensive and the nucleoside polyphosphates that solvability is poor as raw material.In addition, because above method reaction yield is not high, by product is many, product separation difficulty.Therefore, set up a kind of efficient and practical method for symmetric double nucleoside polyphosphates compound and there is very large actual application value.
Summary of the invention
The object of the invention is for the chemosynthesis of symmetric double nucleoside polyphosphates (triphosphoric acid, four phosphoric acid and five phosphoric acid) sodium salt provides a kind of novel, practical and efficient method.
As shown in Figure 1, the symmetric double ribonucleoside triphosphote that the present invention relates to, four phosphoric acid and five phosphoric acid novel method for synthesizing are with 4,5-dicyano imidazole (DCI) is activating reagent, respectively with monophosphate, pyrophosphate salt and triphosphate for nucleophilic reagent, with more than twice excessive nucleosides phosphinylidyne piperidines triethylamine salt (
1-
4) after coupling generates corresponding nucleoside polyphosphates intermediate, continue react the double-core guanosine triphosphate, four phosphoric acid and five phosphoric acid product that generate symmetry with remaining nucleosides phosphinylidyne piperidines in one pot reaction.Crude product through ion-exchange chromatogram purification obtain 4 kinds of double-core guanosine triphosphates (
5
–8
), 3 kinds of dinucleoside tetraphosphates (
9
–11
) and 2 kinds of double-core glycosides five phosphoric acid (
12
–13
).
In present method, the monophosphate of use can be two (three positive fourth ammoniums) salt, two (TBuA) salt, two (three just pungent ammoniums) salt; Pyrophosphate salt can be three (three second ammoniums) salt, three (three positive fourth ammoniums) salt, three (TBuA) salt, three (three just pungent ammoniums) salt; Triphosphate can be three (three positive fourth ammoniums) salt, three (TBuA) salt, three (three just pungent ammoniums) salt.
Nucleosides phosphinylidyne piperidines and monophosphate/pyrophosphate salt/triphosphate and 4, the molar ratio of 5-dicyano imidazole is 1:0.3:2.5 to 1:0.5:3.0, i.e. nucleosides phosphinylidyne piperidines: monophosphate: the molar ratio of 4,5-dicyano imidazole is 1:0.3:3.0; Nucleosides phosphinylidyne piperidines: pyrophosphate salt: the molar ratio of 4,5-dicyano imidazole is 1:2.5 to 1:3.0; Nucleosides phosphinylidyne piperidines: triphosphate: the molar ratio of 4,5-dicyano imidazole is 1:0.5:3.0.Reaction solvent is anhydrous
n,
n-dimethyl formamide (DMF) or
n-crassitude (NMP); Temperature of reaction 20
oc; 20 to 24 hours reaction times.In present method last handling process, first obtain sodium salt crude product through alcohol settling, then obtain double-core glycosides Tripyrophosphoric acid ammonium salt through ion-exchange gel (DEAE-A25) column chromatography.Finally, after sodium form 732 Zeo-karb, symmetric double nucleoside polyphosphates sodium salt is obtained.
Compared with the method reported with previous literature, patent of the present invention only uses the nucleosides phosphinylidyne piperidines that solvability is good, reactive behavior is high to be raw material, utilize 4,5-dicyano imidazole is activating reagent, by controlling the consumption of monophosphate, tetra-sodium or triphosphoric acid, one pot of coupling generates corresponding two nucleoside polyphosphates, avoids and uses nucleoside polyphosphates raw material, significantly simplify synthetic method, product separation productive rate can reach 31 – 62% simultaneously.The method is that the synthesis of symmetric double nucleoside polyphosphates sodium salt establishes a kind of efficient and general novel method.
Accompanying drawing explanation
Fig. 1. the novel method for synthesizing of symmetric double ribonucleoside triphosphote, four phosphoric acid and five sodium phosphate salts.
Embodiment
Below in conjunction with composition graphs 1, the present invention is further elaborated.
embodiment 1:
Two cytidine triphosphate(CTP) trisodium salt (
6) synthesis: by cytidine 5 '-phosphinylidyne piperidines triethylammonium salts (
2, 100 mg, 0.20 mmol), monophosphate two (three positive fourth ammoniums) salt (24 mg, 0.08 mmol) and 4,5-dicyano imidazole (59 mg, 0.50 mmol) be dissolved in dry DMF(2 mL), 20
oc reacts 24 hours.Concentration of reaction solution, adds 3 M sodium acetate aqueous solutions (0.5 mL) in residue, then adds ethanol (50 mL).Centrifugally obtain two cytidine triphosphate(CTP) sodium salt crude product.Ion-exchange gel column chromatography, with 0.3 – 0.6 M ammonium bicarbonate buffer solution linear elution, merges containing product liquid elutriant, and freeze-drying obtains two cytidine triphosphate(CTP) ammonium salt.After sodium form 732 Zeo-karb two cytidine triphosphate(CTP) trisodium salt (
6) 36 mg, productive rate 58%.
embodiment 2:
Two adenosine triphosphate trisodium salt (
7) synthesis: by adenosine 5 '-phosphinylidyne piperidines triethylammonium salts (
3, 100 mg, 0.19 mmol), monophosphate two (TBuA) salt (45 mg, 0.08 mmol) and 4,5-dicyano imidazole (56 mg, 0.48 mmol) be dissolved in dry DMF(2 mL), 20
oc reacts 24 hours.Concentration of reaction solution, adds 3 M sodium acetate aqueous solutions (0.5 mL) in residue, then adds ethanol (50 mL).Centrifugally obtain two adenosine triphosphate acid sodium-salt crude product.Ion-exchange gel column chromatography, with 0.3 – 0.6 M ammonium bicarbonate buffer solution linear elution, merges containing product liquid elutriant, and freeze-drying obtains two adenosine triphosphate acid ammonium salt.Obtain after sodium form 732 Zeo-karb two adenosine triphosphate trisodium salt (
7) 38 mg, productive rate 60%.
embodiment 3:
Two uridine four tetrasodium phosphate salt (
9) synthesis: by uridine 5 '-phosphinylidyne piperidines triethylammonium salts (
1, 100 mg, 0.20 mmol), tetra-sodium three (TBuA) salt (72 mg, 0.08 mmol) and 4,5-dicyano imidazole (71 mg, 0.60 mmol) be dissolved in dry DMF(2 mL), 20
oc reacts 20 h.Concentration of reaction solution, adds 3 M sodium acetate aqueous solutions (0.5 mL) in residue, then adds ethanol (50 mL).Centrifugally obtain two uridine sodium tetraphosphate salt crude product.Ion-exchange gel column chromatography, with 0.3 – 0.6 M ammonium bicarbonate buffer solution linear elution, merges containing product liquid elutriant, and freeze-drying obtains two uridine four ammonium phosphate salt.Obtain after sodium form 732 Zeo-karb two uridine four tetrasodium phosphate salt (
9) 41 mg, productive rate 58%.
embodiment 4:
Two guanosine tetraphosphate(ppGpp) tetra-na salt (
11) synthesis: by guanosine 5 '-phosphinylidyne piperidines triethylammonium salts (
4, 100 mg, 0.19 mmol), tetra-sodium three (three positive fourth ammoniums) salt (59 mg, 0.08 mmol) and 4,5-dicyano imidazole (67 mg, 0.57 mmol) be dissolved in dry DMF(2 mL), 20
oc reacts 20 hours.Concentration of reaction solution, adds 3 M sodium acetate aqueous solutions (0.5 mL) in residue, then adds ethanol (50 mL).Centrifugally obtain two uridine triphosphate sodium salt crude product.Ion-exchange gel column chromatography, with 0.3 – 0.6 M ammonium bicarbonate buffer solution linear elution, merges containing product liquid elutriant, and freeze-drying obtains two uridine triphosphate ammonium salt.Obtain after sodium form 732 Zeo-karb two guanosine tetraphosphate(ppGpp) tetra-na salt (
11) 38 mg, productive rate 52%.
embodiment 5:
Two adenosine five phosphoric acid five sodium-salt (
12) synthesis: by adenosine 5 '-phosphinylidyne piperidines triethylammonium salts (
3, 100 mg, 0.19 mmol), triphosphoric acid three (three positive fourth ammoniums) salt (54 mg, 0.07 mmol) and 4,5-dicyano imidazole (67 mg, 0.57 mmol) be dissolved in dry NMP(2 mL), 20
oc reacts 24 hours.After reaction terminates, in NMP, directly add 3 M sodium acetate aqueous solutions (0.5 mL), then add ethanol (50 mL).Centrifugally obtain two adenosine five sodium phosphate salt crude product.Ion-exchange gel column chromatography, with 0.5 – 0.9 M ammonium bicarbonate buffer solution linear elution, merges containing product liquid elutriant, and freeze-drying obtains two adenosine five ammonium phosphate salt.Obtain after sodium form 732 Zeo-karb two adenosine five phosphoric acid five sodium-salt (
12) 21 mg, productive rate 35%.
embodiment 6:
Two guanosine pentaphosphate five sodium-salt (
13) synthesis: by guanosine 5 '-phosphinylidyne piperidines triethylammonium salts (
4, 100 mg, 0.19 mmol), tetra-sodium three (TBuA) salt (65 mg, 0.07 mmol) and 4,5-dicyano imidazole (67 mg, 0.57 mmol) be dissolved in dry NMP(2 mL), 20
oc reacts 24 hours.After reaction terminates, in NMP, directly add 3 M sodium acetate aqueous solutions (0.5 mL), then add ethanol (50 mL).Centrifugally obtain two guanosine pentaphosphate sodium salt crude product.Ion-exchange gel column chromatography, with 0.5 – 0.9 M ammonium bicarbonate buffer solution linear elution, merges containing product liquid elutriant, and freeze-drying obtains two guanosine pentaphosphate ammonium salt.Obtain after sodium form 732 Zeo-karb two guanosine pentaphosphate five sodium-salt (
13) 20 mg, productive rate 31%.
Claims (5)
1. one kind is synthesized the method for symmetric double ribonucleoside triphosphote, four phosphoric acid and five sodium phosphate salts, it is characterized in that comprising the steps: with 4,5-dicyano imidazole is activating reagent, respectively with monophosphate, pyrophosphate salt and triphosphate for nucleophilic reagent, after generating corresponding nucleoside polyphosphates intermediate to more than twice excessive nucleosides phosphinylidyne piperidines triethylamine salt coupling, continue to react to remaining nucleosides phosphinylidyne piperidines in one pot reaction to generate corresponding symmetric double ribonucleoside triphosphote, four phosphoric acid and five sodium phosphate salts.
2. method according to claim 1, is characterized in that described monophosphate is two (three positive fourth ammoniums) salt, two (TBuA) salt, or two (three just pungent ammoniums) salt; Described pyrophosphate salt is three (three second ammoniums) salt, three (three positive fourth ammoniums) salt, three (TBuA) salt, or three (three just pungent ammoniums) salt; Described triphosphate is three (three positive fourth ammoniums) salt, three (TBuA) salt, or three (three just pungent ammoniums) salt.
3. method according to claim 1, is characterized in that nucleosides phosphinylidyne piperidines: monophosphate: the molar ratio of 4,5-dicyano imidazole is 1:0.3:3.0; Nucleosides phosphinylidyne piperidines: pyrophosphate salt: the molar ratio of 4,5-dicyano imidazole is 1:2.5 to 1:3.0; Nucleosides phosphinylidyne piperidines: triphosphate: the molar ratio of 4,5-dicyano imidazole is 1:0.5:3.0.
4. method according to claim 1, is characterized in that reaction solvent is anhydrous
n,
n-dimethyl formamide or
n-crassitude; Temperature of reaction 20
oc; 20 to 24 hours reaction times.
5. method according to claim 1, after it is characterized in that generating corresponding symmetric double ribonucleoside triphosphote, four phosphoric acid and five sodium phosphate salts, first obtain sodium salt crude product through alcohol settling, obtain corresponding double-core glycosides Tripyrophosphoric acid ammonium salt through ion-exchange gel DEAE-A25 chromatography again, obtain corresponding symmetric double ribonucleoside triphosphote, four phosphoric acid and five sodium phosphate salts finally by after sodium form 732 Zeo-karb.
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| CN201410404196.0A CN104262438A (en) | 2014-08-18 | 2014-08-18 | Method for synthesizing symmetric bi-ribonucleoside triphosphate sodium salt, bi-ribonucleoside tetraphosphate sodium salt and bi-ribonucleoside pentaphosphates odium salt |
| CN201510018937.6A CN104592338B (en) | 2014-08-18 | 2015-01-15 | A kind of method for synthesizing symmetric double ribonucleoside triphosphote, four phosphoric acid and five sodium ascorbyl phosphates |
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|---|---|---|---|
| CN201410404196.0A CN104262438A (en) | 2014-08-18 | 2014-08-18 | Method for synthesizing symmetric bi-ribonucleoside triphosphate sodium salt, bi-ribonucleoside tetraphosphate sodium salt and bi-ribonucleoside pentaphosphates odium salt |
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| CN201510018937.6A Expired - Fee Related CN104592338B (en) | 2014-08-18 | 2015-01-15 | A kind of method for synthesizing symmetric double ribonucleoside triphosphote, four phosphoric acid and five sodium ascorbyl phosphates |
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| WO2008024169A1 (en) * | 2006-07-21 | 2008-02-28 | Glsynthesis, Inc. | Reactive pyrophosphoric and bisphosphonic acid derivatives and methods of their use |
| WO2008012949A1 (en) * | 2006-07-26 | 2008-01-31 | Yamasa Corporation | Process for producing di(pyrimidine nucleoside 5'-)polyphosphate |
| CN103788140A (en) * | 2014-01-26 | 2014-05-14 | 江西科技师范大学 | Method for synthesizing nucleoside 5'-beta, gamma methylene and dihalogenated methylene triphosphate |
| CN103910774A (en) * | 2014-04-09 | 2014-07-09 | 江西科技师范大学 | Method for synthesizing dinucleoside diphosphate and dinucleoside triphosphote |
| CN103923143B (en) * | 2014-04-10 | 2017-01-11 | 江西科技师范大学 | Method for synthesizing dinucleoside tetraphosphate and P2,P3-(dihalo) methylene dinucleoside tetraphosphate |
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| CN104592338B (en) | 2017-09-19 |
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Application publication date: 20150107 |