CN100537581C - Cyclic petroleum phosphate compound and its preparation - Google Patents

Cyclic petroleum phosphate compound and its preparation Download PDF

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CN100537581C
CN100537581C CNB2004100478876A CN200410047887A CN100537581C CN 100537581 C CN100537581 C CN 100537581C CN B2004100478876 A CNB2004100478876 A CN B2004100478876A CN 200410047887 A CN200410047887 A CN 200410047887A CN 100537581 C CN100537581 C CN 100537581C
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methyl
oxyethyl group
xanthoglobulin
phosphoryl
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CN1709898A (en
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张礼和
古险峰
杨振军
张亮仁
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Peking University
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Abstract

The present invention discloses a circle pyrophosphate ester compound and a preparation method thereof. The present invention aims to provide a cADPR analogue with strong stability and high activity, and a preparation method thereof. The circle pyrophosphate ester compound is in the structure of the general formula (I); in the general formula (I), X is C or O; Y is C or O. The circle pyrophosphate ester compound is prepared by taking N<9>-[(tertiary butyl dimethyl silicone base-O-ethoxyl)-methyl]-hypoxanthine as raw materials. The cIDPRE is a cADPR analogue with membrane permeability and has the characteristics of simple and convenient synthetic method, little limitation to substrates, large yield of synthetic products, etc. The product has the advantages of high stability, wide decorative range, and the like and is an ideal pharmacology tool; the product can emit fluorescent light and can be used as materials at various aspects of the research of the calcium ion release of calcium library in cells guided by cADPR, calcium signal conduction in cells participated by cADPR.

Description

Ring-focus phosphate compounds and preparation method thereof
Technical field
The present invention relates to ring-focus phosphate compounds and preparation method thereof.
Background technology
Ring gland glycosides bisphosphate ribose (cADPR) is endogenic intracellular calcium agonist, and Recent study shows that it can also be as the second messenger of signal conduction in the cell.Because cADPR is easy acidolysis and enzymolysis in cell, thereby it is strong to press for stability, active high cADPR analogue is studied at intracellular mechanism of action cADPR as pharmacological tool.The method of at present synthetic cADPR analogue mainly contains three kinds: enzyme process is synthetic, and biomimetic method is synthetic and chemical method is synthetic.Wherein enzyme process is synthetic synthetic bigger to the limitation of substrate with biomimetic method.
Summary of the invention
The purpose of this invention is to provide ring-focus phosphate compounds that stability is strong, activity is high and preparation method thereof.
Ring-focus phosphate compounds provided by the present invention is the compound with general formula (I),
Figure C200410047887D00041
X is CH in the formula 2Or O, Y is CH 2Or O.
The preparation method of the ring-focus phosphate compounds of general formula (I) expression comprises the steps: 1) with N 9-[(tertiary butyl dimethyl silica-based-O-oxyethyl group)-methyl]-xanthoglobulin is dissolved in the methylene dichloride, add DBU after, drip chloromethane ethoxyacetic acid ethyl ester, stir solvent evaporated behind 0.5-1h under the room temperature, column chromatography gets N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-(methyl-ethylene glycol)-xanthoglobulin;
2) with N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-after [(tertiary butyl dimethyl silica-based-O-oxyethyl group)-methyl]-xanthoglobulin is dissolved in THF, add the THF solution of TBAF, stir solvent evaporated behind 1-3h under the room temperature, column chromatography gets N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-(methyl-ethylene glycol)-xanthoglobulin;
3) with N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-(methyl-ethylene glycol)-xanthoglobulin is dissolved in the anhydrous pyridine, adds 1-H tetrazolium and (PhNH) 2POCl stirs solvent evaporated behind 44-52h under the room temperature, column chromatography gets N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin;
4) with N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin is dissolved in the anhydrous methanol, splashes into sodium methoxide solution, and stirring at room 1-3h filters the back concentrated filtrate, gets N 1-[methyl-ethylene glycol]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin;
5) with N1-[methyl-ethylene glycol]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin is dissolved in the anhydrous pyridine, adds TPSCl (2,4; the 6-triisopropylphenylsulfonyl chloride), 1-H tetrazolium and PSS (S, S '-hexichol sulfydryl phosphoryl cyclohexylamine salt); stirring at room 20-28h, solvent evaporated, column chromatography gets N 1-[[(phenylbenzene sulfo-) phosphoryl-O-oxyethyl group]-methyl]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin;
6) with N 1-[[(phenylbenzene sulfo-) phosphoryl-O-oxyethyl group]-methyl]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin is dissolved in pyridine-acetate-diacetyl oxide, adds Isopentyl nitrite, and stirring at room 6-10h is dissolved in after the solvent evaporated in the anhydrous pyridine, adds H 3PO 2With stir 10-12h under the triethylamine room temperature, solvent evaporated, the extraction, column chromatography, after the freeze-drying N 1-[(phenyl thiophosphoryl base-O-oxyethyl group)-methyl]-N 9-[(phosphoryl-O-oxyethyl group)-methyl]-hypoxanthic triethylamine salt;
7) will activate well
Figure C200410047887D0005141515QIETU
Molecular sieve and I 2Place reaction flask, add anhydrous pyridine, stir under the room temperature, N 1-[(phenyl thiophosphoryl base-O-oxyethyl group)-methyl]-N 9The triethylamine salt of-[(phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin (formula (XII)) injects above-mentioned reaction flask, continues to stir 1-3 hours, filters out molecular sieve, gets N after extraction, column chromatography, the freeze-drying 1-[(phosphoryl-O-oxyethyl group)-methyl]-N 9The triethylamine salt of-[(phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin-ring-focus phosphate.
Wherein said N 9-[(tertiary butyl dimethyl silica-based-O-oxyethyl group)-methyl]-xanthoglobulin prepares as follows: 1) VITAMIN B4 is dissolved in the anhydrous pyridine, added behind the Benzoyl chloride back flow reaction 1-2 hours, solvent evaporated, is filtered and is concentrated extraction, N 6-benzoyl-VITAMIN B4;
2) N6-benzoyl-VITAMIN B4 is dissolved in anhydrous 1,2 ethylene dichloride, adds N, the two trimethyl silicane yl acetamides (BSA) of O-are heated to 110-130 ℃, add chloro methoxyacetic acid ethyl ester, at 70-90 ℃ of following stirring reaction 1-2h, extraction, concentrate N 6-benzoyl-N 9-[(acetyl-O-oxyethyl group)-methyl]-VITAMIN B4;
3) with N 6-benzoyl-N 9-[(acetyl-O-oxyethyl group)-methyl]-VITAMIN B4 is dissolved in methyl alcohol-ammoniacal liquor, stirring reaction 20-28h under the room temperature, and solvent evaporated, the water recrystallization gets N 9-methyl glycol-VITAMIN B4;
4) with N 9-methyl glycol-VITAMIN B4 is dissolved in Glacial acetic acid, stirs to add sodium nitrite in aqueous solution down, stirs 20-28h under the room temperature, and solvent evaporated gets N 9-methyl glycol-xanthoglobulin;
5) with N 9-methyl glycol-xanthoglobulin is dissolved in the dry DMF, adds imidazoles and TERT-BUTYL DIMETHYL CHLORO SILANE, stirs 2-4h under the room temperature, through precipitation, filtration and concentrated, gets N 9-[(tertiary butyl dimethyl silica-based-O-oxyethyl group)-methyl]-xanthoglobulin.
Building-up reactions formula of the present invention is as follows, and reaction formula (1) is the synthetic N of VITAMIN B4 9-[(tertiary butyl dimethyl silica-based-O-oxyethyl group)-methyl]-hypoxanthic reaction formula; Reaction formula (2) is with N 9The synthetic N of-[(tertiary butyl dimethyl silica-based-O-oxyethyl group)-methyl]-xanthoglobulin 1-[(phosphoryl-O-oxyethyl group)-methyl]-N 9The reaction formula of-[(phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin-ring-focus phosphate.
Figure C200410047887D00061
Reagent: (a) BzCl, Py; (b) BSA, TMSOTf, DCE; (c) NH 3/ CH 3OH; (d) NaNO 2, AcOH; (e) TBDMSCl, Imidazole, DMF.
Figure C200410047887D00062
Reagent: (a) DBU, ClCH 2CH 2OAC, CH 2Cl 2(b) TBAF, THF; (c) (PhNH) 2POCl, tetrazole, Py; (d) CH 3Ona, CH 3OH; (e) PSS, TPSCl, tetrazole, Py; (f) iisoamyl nitrite, Py:ACOH:AC 2O (2:1:1), ii H 3PO 2, Et 3N, Py; (g) I 2,
Figure C200410047887D0005141515QIETU
MS, Py.
Characteristics such as cIDPRE of the present invention is the cADPR analogue of membrane permeability, has simple synthetic method, and is little to the substrate limitation, that the synthetic product amount is big; Advantages such as product self can send fluorescence, and good stability, modification scope are wide, the calcium ion that can be used as calcium storehouse in the research cADPR mediated cell discharges, and the material of the each side such as intracellular calcium signal conduction that cADPR participates in is a kind of ideal pharmacological tool.
Embodiment
Embodiment 1, N 1-[(phosphoryl-O-oxyethyl group)-methyl]-N 9-[(phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin-ring-focus phosphate synthetic (cIDPDE, X=O in the formula (I), Y=O)
All solvents, raw material and reagent are analytical pure or chemical pure as not specializing.The no water treatment of solvent is carried out according to ordinary method.
Instrument and method that product separates, identifies: tlc silica gel GF 254, column chromatography silochrom (200-300 order), silica gel H be Haiyang Chemical Plant, Qingdao and produce; TLC develops the color by the 254nm ultraviolet detection or with 5% phosphomolybdic acid ethanol solution; Fusing point uses XT-4A type fusing point instrument to measure, and thermometer is not proofreaied and correct; Optically-active uses Perkin-Elmer 243B polarimeter to measure; Infrared spectra uses DE-983G determination of infrared spectroscopy, pressing potassium bromide troche; The syrup liquid-film method.FAB and MALDI-TOF are by VG-ZAB-HS and Bruker APEX TMII, HR-FAB uses Bruker BIFLEX TMThe III mass spectrograph is measured; UV spectrum is used Pharmacia LKB Biochrom 4060 spectrophotometric determinations.Nuclear magnetic resonance spectrum uses Varian VXR-500, JEOL AL300, Bruker Advance 300 nmr determinations, and hydrogen spectrum, carbon spectrum are interior mark with TMS; The phosphorus spectrum is with 85%H 3PO 4Be external standard; Ultimate analysis uses the PE-240C elemental analyser to measure.74900 types that syringe pump is produced with Cole-Parmer company.HPLC uses Gilson high performance liquid phase instrument, uses Delter Paker C-18 semipreparative column to separate.
1, synthetic N 6-benzoyl-VITAMIN B4 (N 6-Benzyl-Adenosine) (formula (II))
(10g 74mmol) is dissolved in the 50ml anhydrous pyridine (Py), stirs to drip Benzoyl chloride (26g down with VITAMIN B4,222mmol), dropwise backflow two hours, stopped reaction, solvent evaporated adds saturated sodium bicarbonate and stirs 30min, uses the chloroform extraction water layer, merge organic phase, anhydrous sodium sulfate drying filters, concentrate, gained crude product chloroform recrystallization gets white needle-like crystals (formula (II)) 14g, productive rate is 79%, and fusing point is 248.5-249.5 ℃.
2, synthetic N 6-benzoyl-N 9-[(acetyl-O-oxyethyl group)-methyl]-VITAMIN B4 (formula (III))
N 6-Benzyl-N 9-[(Acetoxy-ethoxy)-methyl]-Adenosine
With N 6-benzoyl-VITAMIN B4 (formula (II)) (5.9g; 24.7mmol) be dissolved in 200ml anhydrous 1; in 2 ethylene dichloride, add N, two trimethyl silicane yl acetamide (the BSA) (12.4ml of O-; 49mmol); be heated to 120 ℃, treat that solid dissolves (approximately 20min) fully, adds chloromethane ethoxyacetic acid ethyl ester (4.9ml; 29.6mmol); stir 1.5h down at 80 ℃, be cooled to room temperature, add the 200ml ethyl acetate; wash with 100 * 2ml saturated sodium bicarbonate; the washing of 100ml saturated common salt, anhydrous sodium sulfate drying concentrates; the normal pressure column chromatography gets white solid (formula (III)) 3.1g, and productive rate is 35%.
1H?NMR(300MHz，DMSO)δ?1.936(s，3H，CH 3CO-)，3.75-3.78(m，2H，H 5′)，4.08-4.11(m，2H，H 4′)，5.71(s，2H，H 1′)，7.53-7.68(m，5H，ArH)，8.64(s，1H，H 2)，8.79(s，1H，H 8)，11.24(s，1H，NHCO)。Anal.Calcd?for?C 17H 17N 5O 4·1/5Et 2O：C?57.75，H?5.17，N?18.92；Found?C?57.20，H?4.94，N?18.92。
Data show that synthetic product is correct.
3, synthetic N 9-methyl glycol-VITAMIN B4 (N 9-(methyl-glycol)-Adenosine) (formula (IV))
With N 6-benzoyl-N 9-[(acetyl-O-oxyethyl group)-methyl]-VITAMIN B4 (formula (III)) (3.1g 8.7mmol) is dissolved in 300ml methyl alcohol-ammoniacal liquor (v:v=1:1), stirs 24h under the room temperature, solvent evaporated, and the water recrystallization gets white solid (formula (IV)) 2.1g, and productive rate is 99%.
1H?NMR(300MHz，DMSO)δ?3.44-3.50(m，4H，H 4′，H 5′)，4.67(t，1H，JOH，CH 2＝5.1，OH)，5.55(s，2H，H 1′)，7.28(s，2H，NH 2)，8.17(s，1H，H 2)，8.27(s，1H，H 8)。
Data show that synthetic product is correct.
4, synthetic N 9-methyl glycol-xanthoglobulin (N 9-(methyl-glycol)-Hypoxantine) (formula V)
With N 9(2.1g 8.6mmol) is dissolved in the 140ml Glacial acetic acid to-methyl glycol-VITAMIN B4 (formula (IV)), and stirring down, the adding Sodium Nitrite gets the aqueous solution (5g+40mlH 2O), emit reddish-brown gas, stir 24h under the room temperature, solvent evaporated, the water recrystallization gets pale yellow crystals (formula V) 1.5g, and productive rate is 71%.
1H?NMR(300MHz，DMSO)δ?3.44-3.50(m，4H，H 4′，H 5′)，4.67(s，1H，OH)，5.55(s，2H，H 1′)，8.07(s，1H，H 2)，8.21(s，1H，H 8)，12.35(s，1H，NH)。Anal.Calcd?for?C 8H 10N 4O 3·1/2H 2O：C?43.84，H?5.06，N?25.56；Found：C，44.43；H，4.774；N，25.99。
Data show that synthetic product is correct.
5, synthetic N 9-[(tertiary butyl dimethyl silica-based-O-oxyethyl group)-methyl]-xanthoglobulin (formula (VI))
N 9-[(TBDMS-O-ethoxy)-methyl]-Hypoxantine
With N 9-methyl glycol-xanthoglobulin (formula V) (942mg, 3.86mmol) be dissolved in the anhydrous N of 25ml, in the dinethylformamide (DMF), add imidazoles (806mg, 5.79mmol) and TERT-BUTYL DIMETHYL CHLORO SILANE (TBDMSCl) (874mg 5.79mmol), stirs 3h under the room temperature, stop to stir, add 30ml H 2O filters, and filter cake washes with water, and filter cake is dissolved in the methylene dichloride, adds anhydrous sodium sulfate drying, filters, and concentrates, and gets white solid (formula (VI)) 912mg.Productive rate is 73%.
1H?NMR(300MHz，DMSO)δ?0.011(s，6H，Si(CH 3) 2)，0.823(s，9H，C(CH 3) 3)，3.56-3.67(m，4H，H 4′，H 5′)，5.57(s，2H，H 1′)，8.10(s，1H，H 2)，8.24(s，1H，H 8)，12.40(s，1H，H 1)； 13C?NMR(75MHz，DMSO)δ?156.62，148.49，146.10，140.68，123.99，72.80，70.53，61.85，25.87，17.90，-5.30。Anal.Calcd?for?C 14H 24N 4O 3Si：C，51.83；H，7.46；N，17.27；Found：C，52.03；H，7.48；N，17.03。
Data show that synthetic product is correct.
6, synthetic N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-[(tertiary butyl dimethyl silica-based-O-oxyethyl group)-methyl]-xanthoglobulin (formula (VII))
N 1-[(Acetoxy-ethoxy)-methyl]-N 9-[(TBDMS-O-ethoxy)-methyl]-Hypoxantine
With N 9(360mg 1.10mmol) is dissolved in the 10ml methylene dichloride-methyl glycol-xanthoglobulin (formula (VI)), adds 1,8-diazabicyclo [5.4.0] undecane-7-alkene (DBU) (0.82ml, 5.50mmol) after, drip chloromethane ethoxyacetic acid ethyl ester (1.65ml, 11.0mmol), stir under the exothermic heat of reaction, room temperature, behind the 30min, TLC detects raw material completely dissolve, solvent evaporated, normal pressure column chromatography (200-300 order silica gel), get yellow oily liquid (formula (VII)) 67mg, productive rate is 61%.
1H?NMR(300MHz，DMSO)δ?0.084(s，6H，Si(CH 3) 2)，0.825(s，9H，C(CH 3) 3)1.96(s，3H，OAc)，3.56-3.67(m，4H，H 4′，H 5′)，3.77(m，2H，H 5′)，4.11(m，2H，H 4′)，5.48(s，2H，H 1′)，5.59(s，2H，H 1′)，8.29(s，1H，H 2)，8.52(s，1H，H 8)； 13C?NMR(75MHz，DMSO)δ?170.20，156.07，149.11，147.80，141.37，123.25，74.77，72.70，70.53，67.03，62.91，61.83，25.82，20.55，17.87，-5.34.Anal.Calcd?forC 19H 32N 4O 6Si：C，51.80；H，7.32；N，12.72；Found：C，51.74，H，7.47；N，12.42。
Data show that synthetic product is correct.
7, synthetic N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-(methyl-ethylene glycol)-xanthoglobulin (formula (VIII))
N 1-[(Acetoxy-ethoxy)-methyl]-N 9-(methyl-glycol)-Hypoxantine
With N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-[(tertiary butyl dimethyl silica-based-O-oxyethyl group)-methyl]-xanthoglobulin (formula (VII)) (268mg, 0.609mmol) be dissolved among the 10mlTHF, THF solution (the 1.25ml that adds 1M tertiary butyl fluoride amine (TBAF), 1.25mmol), stirring under the room temperature, TLC detects the raw material completely dissolve behind the 2h, solvent evaporated, normal pressure column chromatography (200-300 order silica gel) gets white blister solid (formula (VIII)) 178mg, and productive rate is 90%.
1H?NMR(300MHz，DMSO)δ?1.94(s，3H，OAc)，3.44-3.53(m，4H，H 4′，H 5′)，3.76(m，2H，H 5′)，4.09(m，2H，H 4′)，4.69(t，1H，J H5′，OH＝5.0Hz，OH)，5.46(s，2H，H 1′)，5.56(s，2H，H 1′)，8.28(s，1H，H 2)，8.50(s，1H，H 8).Anal.Calcd?for?C 13H 18N 4O 6C，47.85；H，5.56；N，17.17；Found：C，48.00；H，5.75；N，16.98。
Data show that synthetic product is correct.
8, synthetic N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin (formula (IX))
N 1-[(Acetoxy-ethoxy)-methyl]-N 9-[(dianilinophosphoryl-O-ethoxy)-methyl]-Hypoxantine
With N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-(methyl-ethylene glycol)-xanthoglobulin (formula (VIII)) (250mg 0.767mmol) is dissolved in the 5ml anhydrous pyridine, add the 1-H tetrazolium (161mg, 2.30mmol) and (PhNH) 2POCl (613mg 2.30mmol), stirs 48h under the room temperature, solvent evaporated, the normal pressure column chromatography (sherwood oil: acetone=2:1) (200-300 order silica gel) yellow foaming material (formula (IX)) 392mg, productive rate is 92%.
1H?NMR(300MHz，DMSO)δ?1.93(s，3H，OAc)，3.72-3.76(m，4H，H 4′，H 5′)，4.07-4.10(m，4H，H 5′，H 4′)，5.46(s，2H，H 1′)，5.55(s，2H，H 1′)，6.77-7.16(m，10H，ArH)，8.05(d，2H，2×NH)，8.25(s，1H，H 2)，8.49(s，1H，H 8)； 13C?NMR(75MHz，DMSO)δ?170.28，156.07，149.23，147.82，141.40，128.75，123.26，120.30，117.23，74.74，72.45，68.16，67.07，63.58，62.95，20.59，20.53； 31P?NMR(D 2O?81MHz，decoupled?with 1H)δ?3.07(s)。Anal.Calcd?for?C 25H 29N 6O 7P：C，53.96；H，5.25；N，15.10；Found：C，53.95；H，5.45；N，14.82。
Data show that synthetic product is correct.
9, synthetic N 1-[methyl-ethylene glycol]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin (formula (X))
N 1-[methyl-glycol]-N 9-[(dianilinophosphoryl-O-ethoxy)-methyl]-Hypoxantine
With N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin (formula (IX)) (200mg; 0.36mmol) be dissolved in the 10ml anhydrous methanol; drip the sodium methoxide solution of two 5.4M; stirring at room 2h; add acidic cation-exchange resin and transfer the extremely neutrality of pH; filter, filtrate concentrates, and obtains product (formula (X)) 180mg.
10, synthetic N 1-[[(phenylbenzene sulfo-) phosphoryl-O-oxyethyl group]-methyl]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin (formula (XI))
N 1-[[[Bis-(Phenylthio)phosphoryl]oxy-ethoxy]-methyl]-N 9-[(dianilinophosphoryl-O-ethoxy)-methyl]-Hypoxantine
With N 1-[methyl-ethylene glycol]-N 9(180mg 0.36mmol) is dissolved in the 10ml anhydrous pyridine-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin (formula (X)), adds 2; 4, and 6-triisopropylphenylsulfonyl chloride (TPSCl) (217mg, 0.72mmol); the 1-H tetrazolium; S, and S '-hexichol sulfydryl phosphoryl cyclohexylamine salt (PSS) (410mg, 1.08mmol); stirring at room 24h; solvent evaporated, normal pressure column chromatography (200-300 order silica gel) gets product (formula (XI)) 221mg, and productive rate is 79%.
1H?NMR(300MHz，DMSO)δ?3.68-3.71(m，2H，H 5′)，3.81(m，2H，H 4′)，4.07(m，2H，H 5′)，4.31(m，2H，H 4′)，5.47(s，2H，H 1′)，5.55(s，2H，H 1′)，6.77-7.53(m，20H，Ar?H)，8.05(d，2H，2×NH)，8.26(s，1H，H 2)，8.50(s，1H，H 8)； 13C?NMR(75MHz，DMSO)δ?156.09，149.23，147.84，141.95，135.09，129.71，128.75，126.17，125.59，123.28，121.35，120.31，117.22，74.76，72.41，68.14，67.91，67.00，63.52； 31P?NMR(D 2O?81?MHz，decoupled?with? 1H)δ?3.07(s)，49.36(s)。Anal.Calcd?for?C 35H 36N 6O 7P 2S 2·2acetone：C，55.03；H，5.41；N，9.39；Found：C，55.55；H，5.62；N，8.99。
Data show that synthetic product is correct.
11, synthetic N 1-[(phenyl thiophosphoryl base-O-oxyethyl group)-methyl]-N 9-[(phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin (formula (XII))
N1-[[[(Phenylthio)phosphoryl]oxy-ethoxy]-methyl]-N 9-[(phosphoryl-O-ethoxy)-methyl]-Hypoxantine
With N 1-[[(phenylbenzene sulfo-) phosphoryl-O-oxyethyl group]-methyl]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin (formula (XI)) (110mg; 0.14mmol) be dissolved in 4ml pyridine-acetate-diacetyl oxide (2:1:1; v:v:v); add Isopentyl nitrite (isoamyl nitrite); stirring at room 8h, solvent evaporated, the pyridine band is done 3 times; be dissolved in the 3ml anhydrous pyridine, add H 3PO 2Stirring reaction 11h under the back room temperature, solvent evaporated adds H then 2O and CH 2Cl 2Each 5ml separatory, water CH 2Cl 25 * 3ml washes, evaporated under reduced pressure water (temperature is less than 30 ℃).Supercarbonate (TEAB) buffered soln that adds the triethylamine of 2ml 1M, evaporate to dryness.Said mixture is dissolved in the TEAB buffered soln of 1ml 0.1M, oppositely C18 post HPLC separates, gradient is 0-40% acetonitrile 0.1MTEAB in the 30min, flow velocity is 5ml/min, the detection wavelength is 260nm, collect the 31min main peak, get the triethylamine salt 61mg of product (formula (XII)) after the freeze-drying, productive rate is 82%.
1H?NMR(500MHz，DMSO)δ?3.72(m，2H，H 5′)，3.86(m，2H，H 4′)，3.92(m，2H，H 5′)，4.11(m，2H，H 4′)，5.52(s，2H，H 1′)，5.65(s，2H，H 1′)，7.17-7.43(m，5H，Ar?H)，8.26(s，1H，H 2)，8.39(s，1H，H 8)； 31P?NMR(D 2O81MHz，decoupled?with 1H)δ?1.02(s)，17.85(s)。
Data show that synthetic product is correct.
12, synthetic N 1-[(phosphoryl-O-oxyethyl group)-methyl]-N 9-[(phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin-ring-focus phosphate (formula (XIII))
N 1-[(phosphoryl-O-ethoxy)-methyl]-N 9-[(phosphoryl-O-ethoxy)-methyl]-Hypoxantine-cyclicpyrophosphate
To activate well (1.5g) molecular sieve and I 2(159mg, 604 μ mol) place round-bottomed flask, add the 50ml anhydrous pyridine, stir under the room temperature, with N 1-[(phenyl thiophosphoryl base-O-oxyethyl group)-methyl]-N 9Triethylamine salt (the 25mg of-[(phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin (formula (XII)); 29.9 μ mol) be dissolved in the 5ml anhydrous pyridine; with injecting above-mentioned reaction flask in the micro-injection pump 20 hours; after injection finishes, continue to stir 2 hours, fall molecular sieve with diatomite filtration; filter cake is washed with a small amount of; evaporate to dryness filtrate adds entry and chloroform separatory, and water is washed 3 times with chloroform; the evaporate to dryness water; the TEAB that adds 2ml1M, evaporate to dryness is dissolved in said mixture the TEAB buffered soln solution of 1ml0.1M; oppositely C18 post HPLC separates; gradient is 0-80% acetonitrile in the 30min, and 0.1MTEAB, flow velocity are 5ml/min; the detection wavelength is 260nm; collect the 30min main peak, get the triethylamine salt 6mg of (formula (XIII)) after the freeze-drying, productive rate is 61%.
1H?NMR(500MHz，DMSO)δ?3.74(m，2H，H 5′)，3.83(m，2H，H 4′)，4.11(m，4H，H 4′，H 5′)，5.64(s，2H，H 1′)，5.71(s，2H，H 1′)，8.23(s，1H，H 2)，8.51(s，1H，H 8)； 31P?NMR(D 2O?81MHz，decoupled?with? 1H)δ-10.09(s)。HRMS(TOF，positive)for?C 13H 19N 5O 12P 2?Calcd，427.0342[(M+1) -]；Found，427.0435。
Data show that synthetic product is correct.
Synthetic (X=O in the formula (I), the Y=CH of embodiment 2, cIDPDE 2)
With N 6-benzoyl-VITAMIN B4 (formula (II)) (5.5g; 22.5mmol) be dissolved in 200ml anhydrous 1; in 2 ethylene dichloride; under argon shield, add BSA (11.2ml; 45mmol), be heated to 80 ℃, treat that solid dissolves (approximately 20min) fully; add chloromethane ethoxyacetic acid ethyl ester (4.5ml; 25.6mmol), stir 1.5h down at 80 ℃, be cooled to room temperature; add the 200ml ethyl acetate; wash the washing of 100ml saturated common salt, anhydrous sodium sulfate drying with 100 * 2ml saturated sodium bicarbonate; concentrate, and the normal pressure column chromatography (ether-acetone=3:1) must white solid N 6-benzoyl-N 9-[(acetyl-O-oxyethyl group)-methyl]-VITAMIN B4 (formula (III)) 2.9g, productive rate is 36%.
Other step obtains formula (I) compound with embodiment 1, X=O wherein, Y=CH 2).
Synthetic (the X=CH in the formula (I) of embodiment 3, cIDPDE 2, Y=O)
With N 9(180mg 0.55mmol) is dissolved in the 20ml methylene dichloride-methyl glycol-xanthoglobulin (formula (VI)), adds DBU (0.82ml, 5.5mmol) after, dropping chloromethane ethoxyacetic acid ethyl ester (0.41ml, 2.75mmol), exothermic heat of reaction, stir under the room temperature, behind the 30min, TLC detects raw material completely dissolve, solvent evaporated, normal pressure column chromatography (sherwood oil: acetone=5:1), get yellow oily liquid N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin (formula (IX)) 128mg, productive rate is 63%.
Other step obtains formula (I) compound, wherein X=CH with embodiment 1 2, Y=O).
Synthetic (the X=CH in the formula (I) of embodiment 4, cIDPDE 2, Y=CH 2)
With N 6-benzoyl-VITAMIN B4 (formula (II)) (5.5g; 22.5mmol) be dissolved in 200ml anhydrous 1; in 2 ethylene dichloride; under argon shield, add BSA (11.2ml; 45mmol), be heated to 80 ℃, treat that solid dissolves (approximately 20min) fully; add chloromethane ethoxyacetic acid ethyl ester (4.5ml; 25.6mmol), stir 1.5h down at 80 ℃, be cooled to room temperature; add the 200ml ethyl acetate; wash the washing of 100ml saturated common salt, anhydrous sodium sulfate drying with 100 * 2ml saturated sodium bicarbonate; concentrate, and the normal pressure column chromatography (ether-acetone=3:1) must white solid N 6-benzoyl-N 9-[(acetyl-O-oxyethyl group)-methyl]-VITAMIN B4 (formula (III)) 2.9g, productive rate is 36%.
With N 9(180mg 0.55mmol) is dissolved in the 20ml methylene dichloride-methyl glycol-xanthoglobulin (formula (VI)), adds DBU (0.82ml, 5.5mmol) after, dropping chloromethane ethoxyacetic acid ethyl ester (0.41ml, 2.75mmol), exothermic heat of reaction, stir under the room temperature, behind the 30min, TLC detects raw material completely dissolve, solvent evaporated, normal pressure column chromatography (sherwood oil: acetone=5:1), get yellow oily liquid N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin (formula (IX)) 128mg, productive rate is 63%.
Other step obtains formula (I) compound, wherein X=CH with embodiment 1 2, Y=CH 2).

Claims (4)

1, the ring-focus phosphate compounds that has general formula (I),
X is CH in the formula 2Or O, Y is CH 2Or O.
2, ring-focus phosphate compounds according to claim 1 is characterized in that: described X and Y are 0.
3, the preparation method of the ring-focus phosphate compounds of the general formula of claim 1 (I) expression comprises the steps: 1) with N 9-[(tertiary butyl dimethyl silica-based-O-oxyethyl group)-methyl]-xanthoglobulin is dissolved in the methylene dichloride, add DBU after, drip chloromethane ethoxyacetic acid ethyl ester, stir solvent evaporated behind 0.5-1h under the room temperature, column chromatography gets N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-(methyl-ethylene glycol)-xanthoglobulin;
2) with N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-after [(tertiary butyl dimethyl silica-based-O-oxyethyl group)-methyl]-xanthoglobulin is dissolved in THF, add the THF solution of TBAF, stir solvent evaporated behind 1-3h under the room temperature, column chromatography gets N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-(methyl-ethylene glycol)-xanthoglobulin;
3) with N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-(methyl-ethylene glycol)-xanthoglobulin is dissolved in the anhydrous pyridine, adds 1-H tetrazolium and (PhNH) 2POCl stirs solvent evaporated behind 44-52h under the room temperature, column chromatography gets N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin;
4) with N 1-[(acetyl-O-oxyethyl group)-methyl]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin is dissolved in the anhydrous methanol, splashes into sodium methoxide solution, and stirring at room 1-3h filters the back concentrated filtrate, gets N 1-[methyl-ethylene glycol]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin;
5) with N 1-[methyl-ethylene glycol]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin is dissolved in the anhydrous pyridine, adds 2,4, the 6-triisopropylphenylsulfonyl chloride, and 1-H tetrazolium and S, S '-hexichol sulfydryl phosphoryl cyclohexylamine salt, stirring at room 20-28h, solvent evaporated, column chromatography gets N 1-[[(phenylbenzene sulfo-) phosphoryl-O-oxyethyl group]-methyl]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin;
6) with N 1-[[(phenylbenzene sulfo-) phosphoryl-O-oxyethyl group]-methyl]-N 9-[(pentanoic phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin is dissolved in pyridine-acetate-diacetyl oxide, adds Isopentyl nitrite, and stirring at room 6-10h is dissolved in after the solvent evaporated in the anhydrous pyridine, adds H 3PO 2Stirring reaction 10-12h under the back room temperature, solvent evaporated gets N after extraction, column chromatography, the freeze-drying 1-[(phenyl thiophosphoryl base-O-oxyethyl group)-methyl]-N 9-[(phosphoryl-O-oxyethyl group)-methyl]-hypoxanthic triethylamine salt;
7) will activate good 3
Figure C200410047887C0003103136QIETU
Molecular sieve and I 2Place reaction flask, add anhydrous pyridine, stir under the room temperature, N1-[(phenyl thiophosphoryl base-O-oxyethyl group)-methyl]-N 9-[(phosphoryl-O-oxyethyl group)-methyl]-hypoxanthic triethylamine salt injects above-mentioned reaction flask, continues to stir 1-3 hours, filters out molecular sieve, extraction, and column chromatography gets N after the freeze-drying 1-[(phosphoryl-O-oxyethyl group)-methyl]-N 9The triethylamine salt of-[(phosphoryl-O-oxyethyl group)-methyl]-xanthoglobulin-ring-focus phosphate.
4, the described preparation method of claim 3 is characterized in that, described N 9-[(tertiary butyl dimethyl silica-based-O-oxyethyl group)-methyl]-xanthoglobulin prepares as follows: 1) VITAMIN B4 is dissolved in the anhydrous pyridine, added behind the Benzoyl chloride back flow reaction 1-2 hours, solvent evaporated, is filtered and is concentrated extraction, N 6-benzoyl-VITAMIN B4;
2) with N 6-benzoyl-VITAMIN B4 is dissolved in anhydrous 1,2 ethylene dichloride, adds N, and the two trimethyl silicane yl acetamides of O-are heated to 110-130 ℃, add chloromethane ethoxyacetic acid ethyl ester, at 70-90 ℃ of following stirring reaction 1-2h, extraction, concentrate N 6-benzoyl-N 9-[(acetyl-O-oxyethyl group)-methyl]-VITAMIN B4;
3) with N 6-benzoyl-N 9-[(acetyl-O-oxyethyl group)-methyl]-VITAMIN B4 is dissolved in methyl alcohol-ammoniacal liquor, stirring reaction 20-28h under the room temperature, and solvent evaporated, the water recrystallization gets N 9-methyl glycol-VITAMIN B4;
4) with N 9-methyl glycol-VITAMIN B4 is dissolved in Glacial acetic acid, stirs to add sodium nitrite in aqueous solution down, stirs 20-28h under the room temperature, and solvent evaporated gets N 9-methyl glycol-xanthoglobulin;
5) with N 9-methyl glycol-xanthoglobulin is dissolved in the dry DMF, adds imidazoles and TERT-BUTYL DIMETHYL CHLORO SILANE, stirs 2-4h under the room temperature, through precipitation, filtration and concentrated, gets N 9-[(tertiary butyl dimethyl silica-based-O-oxyethyl group)-methyl]-xanthoglobulin.
CNB2004100478876A 2004-06-18 2004-06-18 Cyclic petroleum phosphate compound and its preparation Expired - Fee Related CN100537581C (en)

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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4670424A (en) * 1983-09-19 1987-06-02 Merck & Co., Inc. Cyclic pyrophosphates of purine and pyrimidine acyclonucleosides

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4670424A (en) * 1983-09-19 1987-06-02 Merck & Co., Inc. Cyclic pyrophosphates of purine and pyrimidine acyclonucleosides

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