CN102816184B - (2-(2-oxygen base-4-thiopyrimidine) oxyethyl group) methyl-phosphorous acid ester derivative and preparation and application thereof - Google Patents
(2-(2-oxygen base-4-thiopyrimidine) oxyethyl group) methyl-phosphorous acid ester derivative and preparation and application thereof Download PDFInfo
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Abstract
The invention discloses one (2-(2-oxygen base-4-thiopyrimidine) oxyethyl group) methyl-phosphorous acid ester derivative and preparation and application thereof.This derivative has the structure shown in general formula I:
wherein, R
1for hydrogen, methyl, allyl group, propargyl or benzyl; R
2for hydrogen, ethyl, trimethylacetic acid methyl, isopropoxy carbonyl oxygen methyl or benzyloxycarbonyloxy methyl; The present invention also provides preparation method and the application of this derivative.It is active that derivative of the present invention has higher suppression HBV, can be used as the preparation of lead compound for Anti-HBV drugs.
Description
Technical field
The present invention relates to one (2-(2-oxygen base-4-thiopyrimidine) oxyethyl group) methyl-phosphorous acid ester derivative and preparation method thereof and application, belong to organic compound synthesis and medical applications technical field.
Background technology
Nucleosides (acid) analogue be clinical application the earliest and the maximum antiviral class medicine of marketed drug, their widespread use alleviates the state of an illness of patient effectively, still plays an important role so far.But due to nucleosides (acid) analogue, generally to have toxicity large, and easily bring out persister during application, and clinical application is very limited.Thus efficient, the low toxicity of research and development and novel nucleoside (acid) analogue not easily producing resistance has very important meaning.
The research of current novel nucleoside acid mono mainly through the transformation to base and ribose moieties, thus changes the character of medicine in absorption, distribution, metabolism and toxicity etc., avoids untoward reaction occurs and increases the activity that medicine resists virus variant.
In base modification, base is introduced a substituent class novel nucleoside such as thiocarbonyl group or sulfide based structural by some bibliographical informations, some of them obtain antiviral, antitumor and anti-microbial activity very well.The nucleoside analog MCC-478(Alamifovir that such as a class of Lily company research and development is novel) very strong inhibit activities is shown to hepatitis B virus, having entered at present the clinical III phase tests, its external activity is about 70 times of lamivudine, is 5 ~ 10 times of adefovir ester.The structure of MCC-478 is as follows, and its constructional feature is with-the NH in thiophenyl class substituted purin base 6
2or-OH, this is and the former obvious difference of ucleosides anti-hbv drug; It preserves the acyclic nucleotide monophosphate structure of Adefovir simultaneously, walk around the first step phosphorylation " bottleneck " in body.In other document also once report after replace the oxygen carbonyl on endogenous nucleotide with thiocarbonyl group, because S atomic radius is greater than O atom, can Van der Waals force by force and between surrounding amino acid, thus can the activity of increase compound.
The structural formula of MCC-478
Based on above-mentioned analysis, the present invention introduces sulphur atom or sulfur-bearing substituting group to increase antiviral activity in pyrimidine bases, the acyclic nucleoside phosphonate analogue of thiopyrimidine that design and synthesis one class is novel, to discovery broad-spectrum high efficacy Anti-HBV drugs significant.
Summary of the invention
For the deficiencies in the prior art, the invention provides one (2-(2-oxygen base-4-thiopyrimidine) oxyethyl group) methyl-phosphorous acid ester derivative and preparation method thereof, the present invention also provides the purposes of above-claimed cpd.
Technical scheme of the present invention is as follows:
1. (2-(2-oxygen base-4-thiopyrimidine) oxyethyl group) methyl-phosphorous acid ester derivative
A kind of (2-(2-oxygen base-4-thiopyrimidine) oxyethyl group) methyl-phosphorous acid ester derivative, has the structure shown in general formula I:
Wherein,
R
1for hydrogen, methyl, allyl group, propargyl or benzyl;
R
2for hydrogen, ethyl, trimethylacetic acid methyl, isopropoxy carbonyl oxygen methyl or benzyloxycarbonyloxy methyl.
Preferably, the compound in above-mentioned general formula I is one of following:
1) (2-(2-oxygen base-4-sulfo--3,4-dihydro-pyrimidin-1(2H)-Ji) oxyethyl group) methyl-phosphorous acid diethyl ester (3a),
2) (2-(2-oxygen base-4-sulfo--3,4-dihydro-pyrimidin-1(2H)-Ji) oxyethyl group) methyl-phosphorous acid (4a),
3) (2-(4-methylthio group-4-sulfo--2 oxopyrimidin-1(2H)-Ji) oxyethyl group) methyl-phosphorous acid diethyl ester (4b),
4) (2-(4-allyl sulfo--2-oxopyrimidin-1(2H)-Ji) oxyethyl group) methyl-phosphorous acid diethyl ester (4c),
5) (2-(2-oxygen base-4-propargyl thiopyrimidine-1(2H)-Ji) oxyethyl group) methyl-phosphorous acid diethyl ester (4d),
6) (2-(4-benzene first sulphur-2-oxopyrimidin-1(2H)-Ji) oxyethyl group) methyl-phosphorous acid diethyl ester (4e),
7) (2-(4-methylthio group-2-oxopyrimidin-1(2H)-Ji) oxyethyl group) methyl-phosphorous acid (5b),
8) (2-(4-allyl sulfenyl-2-oxopyrimidin-1(2H)-Ji) oxyethyl group) methyl-phosphorous acid (5c),
9) (2-(2-oxygen base-4-propargyl pyrimidine-1(2H)-Ji) oxyethyl group) methyl-phosphorous acid (5d),
10) (2-(2-oxygen base-4-thiophenyl pyrimidine) oxyethyl group) methyl-phosphorous acid (5e),
11) (2-(4-methylthio group-2-oxopyrimidin) methyl) phosphoryl dioxygen for the two dimethyl propylene acid esters (6b1) of dimethylene-2,2-,
12) ((2-(4-methylthio group-2-oxopyrimidin-1(2H)-Ji) methyl) phosphoryl dioxygen for dimethylene propylene carbonate (6b2),
13) ((2-(4-methylthio group-2-oxopyrimidin-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for dimethylene benzylcarbonate (6b3),
14) ((2-(4-allyl sulfenyl-2-oxopyrimidin-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for the two dimethyl propylene acid esters (6c1) of dimethylene-2,2-,
15) ((2-(4-allyl sulfenyl-2-oxopyrimidin-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for dimethylene propylene carbonate (6c2),
16) ((2-(4-allyl sulfenyl-2-oxopyrimidin-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for dimethylene benzylcarbonate (6c3),
17) 2-(2-oxygen base-(4-alkynes rosickyite yl pyrimidines-1(2H)-Ji) oxyethyl group) methyl) and phosphoryl dioxygen for the two dimethyl propylene acid esters (6d1) of dimethylene-2,2-,
18) ((2-(2-oxygen base-4-alkynes rosickyite yl pyrimidines-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for dimethylene propylene carbonate (6d2),
19) ((2-(2-oxygen base-4-alkynes rosickyite yl pyrimidines-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for dimethylene carbonic acid benzyl (6d3),
20) ((2-(4-benzylthio-2-oxygen base-pyrimidine-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for the two dimethyl propylene acid esters (6e1) of dimethylene-2,2-,
21) ((2-(4-benzylthio-2-oxygen base-pyrimidine-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for dimethylene propylene carbonate (6e2) or
22) ((2-(4-benzylthio-2-oxygen base-pyrimidine-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen is for dimethylene benzylcarbonate (6e3).
2. the preparation method of (2-(2-oxygen base-4-thiopyrimidine) oxyethyl group) methyl-phosphorous acid ester derivative
Take uridylic as starting raw material, key intermediate diethyl (2-(2 is shortened into [(2-chloroethoxy) methyl] diethyl phosphonate under alkali effect, 4-dioxy-1,4-dihydro-1 (2H)-Ji) oxyethyl group) methyl esters 2a, form target compound 3a via thiophosphoric anhydride sulfo-, hydrolysis obtains target compound 4a; Compound 3a and methyl iodide, allyl bromide 98, propargyl bromide or bromotoluene condensation obtain target compound 4b, 4c, 4d or 4e, obtain target compound 5b, 5c, 5d or 5e via hydrolysis, then obtain target compound 6b1-6b3,6c1-6c3,6d1-6d3 or 6e1-6e3 respectively at trimethylacetic acid methyl esters, chloromethyl base propylene carbonate or the condensation of chloromethyl benzylcarbonate;
Synthetic route is as follows:
Reagent and condition: (i) salt of wormwood, DMF (DMF); (ii) pyridine, thiophosphoric anhydride, 120 DEG C, 2h; (iii) acetone, salt of wormwood, backflow; (iv) acetonitrile, bromotrimethylsilane; (v) acetonitrile, bromotrimethylsilane (vi) DMF (DMF), triethylamine.
Concrete operation step is as follows:
(1) intermediate (2-(2,4-dioxy-Isosorbide-5-Nitrae-dihydro-1(2H)-Ji) oxyethyl group) synthesis of methyl acid phosphate diethyl ester (2a)
By 10g uridylic and the anhydrous N of 200mL, N-dimethylformamide (DMF) joins in 500mL round-bottomed flask, 120 ° of C reflux 1 are little all dissolves up to uridylic, add 6.2g salt of wormwood again, 0.48g sodium methylate and 20.5g [(2-chloroethoxy) methyl] diethyl phosphonate, react 8h under 120 ° of C conditions; TLC detection reaction is complete, is evaporated to pasty state, adds about 150mL methylene dichloride, elimination insolubles, and filtrate concentrates, and upper silica gel column chromatography is separated, and moving phase selects volume ratio 30:1 methylene dichloride-methyl alcohol, and obtain colorless oil 2a, productive rate is 34%.
(2) (2-(2-oxo-4-sulfo--3,4-dihydro-pyrimidin-1(2H)-Ji) oxyethyl group) synthesis of methyl-phosphorous acid diethyl ester (3a)
The 2a of 5g is dissolved in 50mL pyridine, is heated to 120 ° of C, react 1 hour, TLC detection reaction reduces pressure steam solvent completely, adds a large amount of frozen water in residue, filters insolubles, solution with ethyl acetate extraction 3 ~ 4 times, organic layer adds anhydrous sodium sulfate drying, filters solvent evaporated and obtains crude product; Crude product silica gel column chromatography is separated, and moving phase is volume ratio 3:1 ethyl acetate-dichloromethane, turns the methylene dichloride of volume ratio 20:1: methyl alcohol, obtains yellow oil 3a.
(3) (2-(2-oxygen base-4-sulfo--3,4-dihydro-pyrimidin-1(2H)-Ji) oxyethyl group) synthesis of methyl-phosphorous acid (4a)
The 3a of 0.5g is dissolved in 7mL acetonitrile, 1mL bromotrimethylsilane (TMSBr) is added under 0 ° of C, reacting at normal temperature without light 18 hours, reaction terminates rear decompression and steams solvent, and 2mol/LNaOH adjusts pH=8,3 times are washed by ethyl acetate, water layer HCl is neutralized to pH=3 ~ 4, cooling, is separated layer oily matter, dry yellow paste 4a in vacuum drying oven, productive rate 71%.
(4) (2-(-4-sulfo--2 oxopyrimidin-dihydro-1(2H)-Ji) oxyethyl group) synthesis of methyl-phosphorous acid diethyl ester (4b-4e)
6mmol2a is dissolved in 15mL acetone, adds 3mmol salt of wormwood and 6mmol methyl iodide, bromopropylene, propargyl bromide or bromotoluene, 80 ° of C back flow reaction 2 hours, TLC detection reaction is complete, and decompression steams solvent, is separated obtains pale yellow oil 4b respectively with silica gel column chromatography, 4c, 4d or 4e.
(5) (2-(4-(methylthio group/allyl sulfenyl/alkynes rosickyite base/thiophenyl)-2-oxopyrimidin-dihydro-1(2H)-Ji) oxyethyl group) synthesis of methyl-phosphorous acid (5b, 5c, 5d or 5e)
According to the synthetic method of 4a, 1g4b, 4c, 4d or 4e and bromotrimethylsilane (TMSBr) react 18 hours in acetonitrile, obtain yellow paste 5b respectively, 5c, 5d or 5e.
(6) (2-(4-methylthio group-2-oxopyrimidin-dihydro-1(2H)-Ji) oxyethyl group) synthesis of methylphosphonate (6b1,6b2 or 6b3)
The 5b of 1mmol is dissolved in the anhydrous DMF of 5mL (DMF), in stirring at room temperature half hour, adds 4mmol triethylamine, continues stirring 1 at room temperature little muddy up to solution.Then 4mmol trimethylacetic acid methyl esters is added, chloromethyl propylene carbonate or chloromethyl benzylcarbonate are warming up to 60 DEG C of reactions 7 hours, TLC detection reaction is complete, add 15mL ethyl acetate and stir 1 hour at 0 ° of C, filter, filtrate evaporate to dryness, be separated with silica gel column chromatography, moving phase is volume ratio 15:1 methylene dichloride-methyl alcohol, obtains faint yellow paste 6b1,6b2 or 6b3.
(7) (2-(4-allyl sulfenyl-2-oxopyrimidin-dihydro-1(2H)-Ji) oxyethyl group) synthesis of methylphosphonate (6c1,6c2 or 6c3)
The 5c of 1mmol is dissolved in the anhydrous DMF of 5mL (DMF), in stirring at room temperature half hour, adds 4mmol triethylamine, continues stirring 1 at room temperature little muddy up to solution.Then 4mmol trimethylacetic acid methyl esters is added, chloromethyl propylene carbonate or chloromethyl benzylcarbonate, be warming up to 60 DEG C of reactions 7 hours, TLC detection reaction is complete, adds 15mL ethyl acetate and stirs 1 hour at 0 ° of C, filter, filtrate evaporate to dryness, be separated with silica gel column chromatography, moving phase is volume ratio 15:1 methylene dichloride-methyl alcohol, obtain faint yellow paste 6c1,6c2 or 6c3.
(8) (2-(4-alkynes rosickyite base-2-oxopyrimidin-dihydro-1(2H)-Ji) oxyethyl group) synthesis of methylphosphonate (6d1,6d2 or 6d3)
The 5d of 1mmol is dissolved in the anhydrous DMF of 5mL (DMF), in stirring at room temperature half hour, adds 4mmol triethylamine, continues stirring 1 at room temperature little muddy up to solution.Then 4mmol trimethylacetic acid methyl esters is added, chloromethyl propylene carbonate or chloromethyl benzylcarbonate, be warming up to 60 DEG C of reactions 7 hours, TLC detection reaction is complete, adds 15mL ethyl acetate and stirs 1 hour at 0 ° of C, filter, filtrate evaporate to dryness, be separated with silica gel column chromatography, moving phase is the methylene chloride-methanol of volume ratio 15:1, obtain faint yellow paste 6d1,6d2 or 6d3.
(9) (2-(4-thiophenyl-2-oxopyrimidin-dihydro-1(2H)-Ji) oxyethyl group) synthesis of methylphosphonate (6e1,6e2 or 6e3)
The 5e of 1mmol is dissolved in the anhydrous DMF of 5mL (DMF), in stirring at room temperature half hour, adds 4mmol triethylamine, continues stirring 1 at room temperature little muddy up to solution.Then 4mmol trimethylacetic acid methyl esters (chloromethylpivalate) is added, chloromethyl propylene carbonate (chloromethylisopropylcarbonate) or chloromethyl benzylcarbonate (benzylchloromethylcarbonate) are warming up to 60 DEG C of reactions 7 hours, TLC detection reaction is complete, add 15mL ethyl acetate and stir 1 hour at 0 ° of C, filter, filtrate evaporate to dryness, be separated with silica gel column chromatography, moving phase is methylene dichloride: methyl alcohol 15:1, obtain faint yellow paste 6e1,6e2 or 6e3.
3. (2-(2-oxygen base-4-thiopyrimidine) oxyethyl group) methyl-phosphorous acid ester derivative pharmaceutical composition
A kind of Anti-HBV drugs composition, containing above-mentioned (2-(2-oxygen base-4-thiopyrimidine) oxyethyl group) methyl-phosphorous acid ester derivative and pharmaceutical excipient, makes the medicine of different dosage form.
4. the application of (2-(2-oxygen base-4-thiopyrimidine) oxyethyl group) methyl-phosphorous acid ester derivative
(2-(3-methylthio group-1,2,4 thiadiazine) oxyethyl group) of the present invention methyl-phosphorous acid ester derivative can be used as the application of HBV nucleoside inhibitor.Specifically, as HBV inhibitor for the preparation of anti-hepatic-B virus medicine.
The present invention is on the basis of Watson-Crick basepairing rule, in conjunction with the constructional feature of lead compound MCC478, the open loop phosphonic structures of ribose moieties in simulation adefovir ester, we have designed and synthesized the novel nucleoside antiviral analogs that a class has brand-new base structure, for avoiding the monophosphate form of nucleosides because electronegativity is difficult to too greatly enter cytolemma, use pivaloyl chloride methyl esters respectively, sec.-propyl carbonic acid chloromethyl ester and benzyl carbonic acid chloromethyl ester and its formation dibasic acid esters prodrug, this compounds has good hepatitis B virus resisting (HBV) activity to have further medicinal Development volue.
Embodiment:
Below in conjunction with embodiment, the present invention will be further described, and the numbering of all target compounds is identical with table 1.Described per-cent number is mass percent.
Embodiment 1: intermediate (2-(2,4-dioxy-Isosorbide-5-Nitrae-dihydro-1(2H)-Ji) oxyethyl group) synthesis of methyl acid phosphate diethyl ester (2a)
By 10g(0.09mol) uridylic and the anhydrous N of 200mL, N-dimethylformamide (DMF) joins in 500mL round-bottomed flask, 120 ° of C reflux 1 are little all dissolves up to uridylic, add 6.2g(0.05mol again) salt of wormwood, 0.48g(9mmol) sodium methylate and 20.5g(0.09mol) [(2-chloroethoxy) methyl] diethyl phosphonate, reacts 8h under 120 ° of C conditions.TLC detection reaction is complete, is evaporated to pasty state, adds about 150mL methylene dichloride, elimination insolubles, filtrate concentrates, and upper silica gel column chromatography is separated, and moving phase selects methylene dichloride: methyl alcohol (30:1), obtain 9.5g colorless oil 2a, productive rate is 34%, EI-MS:m/z307.4(M+H
+), 329.5(M+Na
+).
Embodiment 2:(2-(2-oxo-4-sulfo--3,4-dihydro-pyrimidin-1(2H)-Ji) oxyethyl group) synthesis of methyl-phosphorous acid diethyl ester (3a)
By 5g(16mmol) 2a is dissolved in 50mL pyridine, be heated to 120 ° of C, react 1 hour, TLC detection reaction reduces pressure steam solvent completely, add a large amount of frozen water in residue, filter insolubles, solution with ethyl acetate extraction 3 ~ 4 times, organic layer adds anhydrous sodium sulfate drying, filters solvent evaporated and obtains crude product.Crude product silica gel column chromatography is separated, and moving phase is ethyl acetate: methylene dichloride (3:1) turns methylene dichloride: methyl alcohol (20:1), obtains about 2.1g yellow oil, and productive rate is 41%, EI-MS:m/z323.6(M+H
+);
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：12.70(s,1H,NH),7.48(d,1H,J=7.2Hz,CH),6.24(dd,1H,CH,J
1=2.1Hz,J
2=7.5Hz),4.01-3.98(m,4H,2-CH
2-CH
3),3.89(t,2H,J=6.0Hz,N-CH
2-CH
2),3.84(d,2H,J=7.8Hz,O-CH
2-P),3.72(t,2H,J=6.3Hz,CH
2-CH
2-O),1.22(t,6H,J=7.2Hz,2-CH
2-CH
3);13CNMR(150MHz,DMSO-d6)δppm：190.7(C=S),148.8(C=O),142.5(N-CH=CH),112.1(CH-C=S),69.8(CH
2-P=O),64.8(CH
2-CH
2-O),62.2(-CH
2-CH
3),48.21(CH
2-CH
2-N),16.7(-CH
2-CH
3).
Embodiment 3:(2-(2-oxygen base-4-sulfo--3,4-dihydro-pyrimidin-1(2H)-Ji) oxyethyl group) synthesis of methyl-phosphorous acid (4a)
0.5g (1.6mmol) 3a is dissolved in 7mL acetonitrile, under 0 ° of C, add 1mL bromotrimethylsilane (TMSBr), reacting at normal temperature without light 18 hours, reaction terminates rear decompression and steams solvent, 2mol/LNaOH adjusts pH ≈ 8, wash three times by ethyl acetate, water layer HCl is neutralized to pH=3 ~ 4, cooling, be separated layer oily matter, the yellow paste of dry 0.3g in vacuum drying oven, productive rate 71%, EI-MS:m/z265.1(M-H);
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：12.80(s,1H,NH),11.27(s,2H,OH),7.51(d,1H,J＝7.2Hz,CH),6.22(dd,1H,CH,J
1=3.0Hz,J
2=7.2Hz),3.84(t,2H,J＝6.3Hz,N-CH2-CH2),3.68(t,2H,J＝6.6Hz,CH2-CH2-O),3.58(d,2H,J＝7.8Hz,O-CH2-P);
Embodiment 4:(2-(4-methylthio group-4-sulfo--2 oxopyrimidin-1(2H)-Ji) oxyethyl group) synthesis of methyl-phosphorous acid diethyl ester (4b)
2g(6mmol) 2a is dissolved in 15mL acetone, add 0.43g(3mmol) salt of wormwood and 6mmol methyl iodide, 80 ° of C back flow reaction 2 hours, TLC detection reaction is complete, and decompression steams solvent, is separated with silica gel column chromatography, moving phase is methylene dichloride: methyl alcohol 20:1, obtain 1.7g pale yellow oil 4b, productive rate is 84%, EI-MS:m/z337.5(M+H
+), 359.3(M+Na
+);
Product spectrum analytical data:
1HNMR (600MHz, DMSO-d6) δ ppm:7.78 (d, 1H, J=7.2Hz; CH), 6.40 (d, 1H, J=7.2Hz; CH), 3.99-3,96 (m, 6H; 2-CH2-CH3 and N-CH2-CH2), 3.76 (d, 2H, J=8.4Hz; O-CH2-P), 2.44 (s, 3H, S-CH3); 1.19 (t, 6H, J=7.2Hz, 2-CH2-CH3); 13CNMR (150MHz, DMSO-d6) δ ppm:176.3 (N=C-S), 154.0 (C=O), 147.5 (N-CH=CH), 102.3 (CH-C-S), 69.8 (CH2-P=O), 64.8 (CH2-CH2-O), 62.2 (-CH2-CH3), 48.5 (CH2-CH2-N), 16.7 (-CH2-CH3), 12.5 (S-CH3).
Embodiment 5:(2-(4-allyl sulfo--2-oxopyrimidin-dihydro-1(2H)-Ji) oxyethyl group) synthesis of methyl-phosphorous acid diethyl ester (4c)
According to the synthetic method of above-mentioned 4b, in 15mL acetone, add 2g2a, 0.43g salt of wormwood and 6mmol bromopropylene, back flow reaction 2 hours, silica gel column chromatography is separated to obtain 1.5g pale yellow oil 4c, and productive rate is 70%, EI-MS:m/z363.4(M+H
+), 385.3(M+Na
+);
Product spectrum analytical data:
1HNMR (600MHz, DMSO-d6) δ ppm:7.80 (d, 1H, J=7.2Hz; CH), 6.40 (d, 1H, J=7.2Hz; CH), 5.89-5.86 (m, 1H ,=CH-CH2); 5.33-5.30 (m, 1H, H-CH=CH), 5.13-5; 10 (m, 1H, H-CH=CH), 3.99-3; 96 (m, 6H, 2-CH2-CH3 and N-CH2-CH2), 3.83 (d; 2H, J=8.4Hz, O-CH2-P), 3.78 (d; 2H, J=6.0Hz ,=CH-CH2), 3.75 (t; 2H, J=7.2Hz, CH2-CH2-O), 1.19 (t; 6H, J=6.6Hz, 2-CH2-CH3); 13CNMR (150MHz, DMSO-d6) δ ppm:175.8 (S-C=N), 153.9 (C=O), 148.0 (N-CH=CH), 133.6 (-CH=CH2), 118.7 (-CH=CH2), 102.4 (CH-C=O), 69.7 (CH2-P=O), 64.8 (CH2-CH2-O), 62.2 (-CH2-CH3), 49.6 (CH2-CH2-N), 31.71 (S-CH2-CH), 16.7 (-CH2-CH3).
Embodiment 6:(2-(2-oxygen base-4-propargyl thiopyrimidine-dihydro-1(2H)-Ji) oxyethyl group) synthesis of methyl-phosphorous acid diethyl ester (4d)
According to the synthetic method of above-mentioned 4b, in 15mL acetone, add 2g2a, 0.43g salt of wormwood and 6mmol propargyl bromide, back flow reaction 2 hours, silica gel column chromatography is separated to obtain about 1.4g pale yellow oil 4c, and productive rate is 64%, EI-MS:m/z362.4(M+2H
+), 383.3(M+Na
+);
Product spectrum analytical data:
1hNMR (600MHz, DMSO-d6) δ ppm:7.86 (d, 1H; J=6.6Hz, CH), 6.44 (d; 1H, J=6.6Hz, CH); 3.99-3,96 (m, 8H; 2-CH2-CH3, N-CH2-CH2 and O-CH2-P), 3.78 (d; 2H, J=3.6Hz, ≡ C-CH2); 3.75 (t, 2H, J=6.0Hz; CH2-CH2-O), 3.20 (t, 1H; J=3.9Hz ,-C ≡ CH) 1.19 (t, 6H; J=7.2Hz, 2-CH2-CH3); 13CNMR (150MHz, DMSO-d6) δ ppm:174.7 (S-C=N), 153.8 (C=O), 148.6 (N-CH=CH), 102.0 (CH-C=O), 80.1 (-C ≡ CH), 74.1 (-C ≡ CH), 69.6 (CH2-P=O), 64.9 (CH2-CH2-O), 62.2 (-CH2-CH3), 49.7 (CH2-CH2-N), 17.5 (S-CH2-C), 16.7 (-CH2-CH3).
Embodiment 7:(2-(4-benzene first sulphur-2-oxopyrimidin-dihydro-1(2H)-Ji) oxyethyl group) synthesis of methyl-phosphorous acid diethyl ester (4e)
According to the synthetic method of above-mentioned 4b, in 15mL acetone, add 2g2a, 0.43g salt of wormwood and 6mmol bromotoluene, back flow reaction 2 hours, silica gel column chromatography is separated to obtain about 1.8g pale yellow oil 4e, and productive rate is 72%.EI-MS：m/z414.4（M+H
+）；
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：7.84(d,1H,J＝7.2Hz,CH),7.41-7.25(m,5H,Ar-H),6.39(d,1H,J＝7.2Hz,CH),4.39(s,2H,Ar-CH2),4.14-4.09（m,4H,2-CH2-CH3）,3.97(t,2H,J＝6.6Hz,N-CH2-CH2),3.75(t,2H,J＝6.3Hz,CH2-CH2-O),3.57(d,2H,J＝8.4Hz,O-CH2-P),1.21(t,6H,J＝7.2Hz,2-CH2-CH3);13CNMR(150MHz,DMSO-d6)δppm：176.5(S-C=N),151.7(C=O),146.7(N-CH=CH),136.2(Ar-C),129.7(Ar-CH),128.5(Ar-CH),127.8(Ar-CH),100.8(CH-C=O),70.3(CH2-P=O),64.8(CH2-CH2-O),62.2(-CH2-CH3),47.4(CH2-CH2-N),31.21(S-CH2-Ar),16.7(-CH2-CH3).
Embodiment 8:(2-(4-methylthio group-2-oxopyrimidin-dihydro-1(2H)-Ji) oxyethyl group) synthesis of methyl-phosphorous acid (5b)
According to the synthetic method of 4a, 1g4b and bromotrimethylsilane (TMSBr) react 18 hours in acetonitrile, obtain the yellow paste (5b) of 0.60g, productive rate 45%, EI-MS:m/z279.2(M-H);
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：11.28(s,2H,OH),7.78(d,1H,J＝7.2Hz,CH),6.40(d,1H,J＝7.2Hz,CH,),4.00(t,2H,J＝6.6Hz,N-CH2-CH2),3.82(d,2H,J＝8.4Hz,O-CH2-P),3.76(t,2H,J＝6.6Hz,CH2-CH2-O);
Embodiment 9:(2-(4-allyl sulfenyl-2-oxopyrimidin-dihydro-1(2H)-Ji) oxyethyl group) synthesis of methyl-phosphorous acid (5c)
According to the synthetic method of 4a, 1g4c and bromotrimethylsilane (TMSBr) react 18 hours in acetonitrile, obtain the yellow paste (5c) of 0.58g, productive rate 54%, EI-MS:m/z305.4(M-H);
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：11.26(s,2H,OH),7.89(d,1H,J＝7.2Hz,CH),6.41(d,1H,J＝7.2Hz,CH),5.92-5.87（m,1H,=CH-CH2）5.33-5.30(m,1H,H-CH=CH),5.14-5.12(m,1H,H-CH=CH),3.96(t,2H,J＝6.3Hz,N-CH2-CH2),3.83(d,2H,J＝8.4Hz,O-CH2-P),3.78(d,2H,J＝6.0Hz,=CH-CH2),3.75(t,3H,J＝6.3Hz,CH2-CH2-O);
Embodiment 10:(2-(2-oxygen base-4-propargyl pyrimidine-dihydro-1(2H)-Ji) oxyethyl group) synthesis of methyl-phosphorous acid (5d)
According to the synthetic method of 4a, 1g4d and bromotrimethylsilane (TMSBr) react 18 hours in acetonitrile, obtain the yellow paste (5d) of 0.55g, productive rate 66%, EI-MS:m/z303.3(M-H);
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：11.27（s,2H,OH）7.96(d,1H,J＝7.2Hz,CH),6.43(d,1H,J＝7.2Hz,CH),5.51(d,2H,J＝3.6Hz,≡C-CH2),3.84(t,2H,J＝6.0Hz,N-CH2-CH2),3.68(t,2H,J＝6.3Hz,CH2-CH2-O),3.57(d,2H,J＝8.4Hz,O-CH2-P),3.40(t,1H,J＝3.9Hz,≡C-H);
Embodiment 11:(2-(2-oxygen base-4-thiophenyl pyrimidine) oxyethyl group) synthesis of methyl-phosphorous acid (5e)
According to the synthetic method of 4a, 1g4e and bromotrimethylsilane (TMSBr) react 18 hours in acetonitrile, obtain the yellow paste (5e) of about 0.56g, productive rate 65%, EI-MS:m/z355.3(M-H);
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：11.26(s,2H,OH),7.84(d,1H,J＝7.2Hz,CH),7.41-7.26（m,5H,Ar-H），6.39(d,1H,J＝7.2Hz,CH),4.39(s,2H,Ar-CH2),3.97(t,2H,J＝6.6Hz,N-CH2-CH2),3.75(t,3H,J＝6.3Hz,CH2-CH2-O),3.57(d,2H,J＝8.4Hz,O-CH2-P);
Embodiment 12:(2-(4-methylthio group-2-oxopyrimidin) methyl) phosphoryl dioxygen is for the synthesis of the two dimethyl propylene acid esters (6b1) of dimethylene-2,2-
5b 0.3g(1mmol) is dissolved in 5mL dry DMF, in stirring at room temperature half hour, adds 0.4g(4mmol) triethylamine, continues stirring 1 at room temperature little muddy up to solution.Then add 0.6g(4mmol) trimethylacetic acid methyl esters (chloromethylpivalate) be warming up to 60 DEG C reaction 7 hours, TLC detection reaction is complete, add 15mL ethyl acetate and stir 1 hour at 0 ° of C, filter, filtrate evaporate to dryness, is separated with silica gel column chromatography, moving phase is methylene dichloride: methyl alcohol 15:1, obtain faint yellow paste 6b1 and be about 0.13g, productive rate is 26%, EI-MS:m/z509.5(M+H
+), 531.3(M+Na
+);
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：7.78(d,1H,J＝7.2Hz,CH),6.41(d,1H,J＝7.2Hz,CH),5.46(d,4H,J＝2.4Hz,2-P-O-CH2),3.97(t,2H,J＝6.0Hz,N-CH2-CH2),3.84(d,2H,J＝7.8Hz,O-CH2-P),3.77(t,3H,J＝6.6Hz,CH2-CH2-O),2.42(s,3H,S-CH3),1.22(s,18H,2-(CH3)3);
Embodiment 13:((2-(4-methylthio group-2-oxopyrimidin-dihydro-1(2H)-Ji) methyl) phosphoryl dioxygen is for the synthesis of dimethylene propylene carbonate (6b2)
According to the synthetic method of compound 6b1, the 5b of 1mmol, 4mmol triethylamine and 4mmol chloromethyl propylene carbonate (chloromethylisopropylcarbonate) are obtained by reacting the faint yellow paste 6b2 of about 0.14g in dry DMF, and productive rate is 27%, EI-MS:m/z513.4(M+H
+), 535.6(M+Na
+);
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：7.78(d,1H,J＝7.2Hz,CH),6.40(d,1H,J＝7.2Hz,CH),5.29(d,4H,J＝2.4Hz,2-P-O-CH2),4.60-4.64(m,2H,2-CH-(CH3)2),3.98(t,2H,J＝6,6Hz,N-CH2-CH2),3.83(d,2H,J＝8.4Hz,O-CH2-P),2.44(s,3H,S-CH3),1.15(d,12H,J＝6.0Hz,2-(CH3)2);
Embodiment 14:((2-(4-methylthio group-2-oxopyrimidin-dihydro-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen is for the synthesis of dimethylene benzylcarbonate (6b3)
According to the synthetic method of compound 6b1, the 5b of 1mmol, 4mmol triethylamine and 4mmol chloromethyl benzylcarbonate (benzylchloromethylcarbonate) are obtained by reacting the faint yellow paste 6b3 of about 0.16g in dry DMF, and productive rate is 26%, EI-MS:m/z609.6(M+H
+), 631.6(M+Na
+);
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：7.79(d,1H,J＝7.2Hz,CH),7.40-7.37(m,5H,H-Ar),6.41(d,1H,J＝6.6Hz,CH),5.30(d,4H,J＝2.4Hz,2-P-O-CH2),4.65(s,4H,2-CH2-Ar),3.98(t,2H,J＝6.6Hz,N-CH2-CH2),3.84(d,2H,J＝8.4Hz,O-CH2-P),3.76(t,2H,J＝6.6Hz,O-CH2-CH2),2.41(s,3H,S-CH3);
Embodiment 15:((2-(4-allyl sulfenyl-2-oxopyrimidin-dihydro-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen is for the synthesis of the two dimethyl propylene acid esters (6c1) of dimethylene-2,2-
According to the synthetic method of compound 6b1, the 5c of 1mmol, 4mmol triethylamine and 4mmol trimethylacetic acid methyl esters are obtained by reacting the faint yellow paste 6c1 of about 0.15g in dry DMF, and productive rate is 28%, EI-MS:m/z535.4(M+H
+), 557.5(M+Na
+);
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：7.80(d,1H,J＝7.2Hz,CH),6.39(d,1H,J＝7.2Hz,CH,),5.90-5.87（m,1H,=CH-CH2）5.33-5.30(m,1H,H-CH=CH),5.23(d,4H,J＝2.4Hz,2-P-O-CH2),5.14-5.12(m,1H,H-CH=CH),3.96(t,6H,J＝6.6Hz,N-CH2-CH2),3.83(d,2H,J＝8.4Hz,O-CH2-P),3.78(d,2H,J＝6.0Hz,=CH-CH2),3.74(t,2H,J＝6.3Hz,CH2-CH2-O),1.18(s,18H,2-(CH3)3);
Embodiment 16:((2-(4-allyl sulfenyl-2-oxopyrimidin-dihydro-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen is for the synthesis of dimethylene propylene carbonate (6c2)
According to the synthetic method of compound 6b2, the 5c of 1mmol, 4mmol triethylamine and 4mmol chloromethyl propylene carbonate are obtained by reacting the faint yellow paste 6c2 of about 0.15g in dry DMF, and productive rate is 28%, EI-MS:m/z539.5(M+H
+);
Product spectrum analytical data:
1HNMR (600MHz, DMSO-d6) δ ppm:7.96 (d, 1H, J=7.2Hz, CH), 6.31 (d, 1H, J=7.2Hz, CH), 5.91-5.87 (m, 1H,=CH-CH2), 5.36-5.29 (m, 3H, H-CH=CH and 2-CH-(CH2) 2), 5.22(d, 4H, J=2.4Hz, P-O-CH2), 5.13-5.11 (m, 1H, H-CH=CH), 3.97 (t, 2H, J=6.3Hz, N-CH2-CH2), 3.78 (d, 2H, J=8.4Hz, O-CH2-P), 3.68 (d, 2H, J=6.0Hz,=CH-CH2), 3.66 (t, 2H, J=6.3Hz, CH2-CH2-O), 1.20 (d, 12H, J=6.6Hz, 2-(CH3) 2),
Embodiment 17:((2-(4-allyl sulfenyl-2-oxopyrimidin-dihydro-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen is for the synthesis of dimethylene benzylcarbonate (6c3)
According to the synthetic method of compound 6b3, the 5c of 1mmol, 4mmol triethylamine and 4mmol chloromethyl benzylcarbonate are obtained by reacting the faint yellow paste 6c3 of about 0.15g in dry DMF, and productive rate is 27%, EI-MS:m/z635.5(M+H
+);
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：7.81(d,1H,J＝7.2Hz,CH),7.40-7.37(m,10H,2H-Ar),6.40(d,1H,J＝6.6Hz,CH),5.92-5.87(m,1H,=CH-CH2),5.32-5.30(m,1H,H-CH=CH),5.25(d,4H,J＝2.4Hz,P-O-CH2),4.54(s,4H,2-CH2-Ar),3.97(t,2H,J＝6.6Hz,N-CH2-CH2),3.83(d,2H,J＝8.4Hz,O-CH2-P),3.78(d,2H,J＝6.0Hz,=CH-CH2),3.74(t,2H,J＝6.6Hz,CH2-CH2-O);
Embodiment 18:2-(2-oxygen base-(4-alkynes rosickyite yl pyrimidines-dihydro-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen is for the synthesis of the two dimethyl propylene acid esters (6d1) of dimethylene-2,2-
According to the synthetic method of compound 6b1, the 5d of 1mmol, 4mmol triethylamine and 4mmol trimethylacetic acid methyl esters are obtained by reacting the faint yellow paste 6d1 of about 0.15g in dry DMF, and productive rate is 28%, EI-MS:m/z533.4(M+H
+), 555.4(M+Na
+);
Product spectrum analytical data:
1HNMR (600MHz, DMSO-d6) δ ppm:8.06 (d, 1H, J=7.2Hz; CH), 6.38 (d, 1H, J=7.2Hz; CH), 5.25 (d, 4H; J=2.4Hz, 2CH2-O-P), 3.97-3.94 (m; 4H, ≡ C-CH2 and O-CH2-P), 3.68 (d; 2H, J=6.6Hz, N-CH2-CH2); 3.35 (t, 2H, J=6.6Hz; CH2-CH2-O), 3.19 (t, 1H; J=3.6Hz, ≡ C-H), 1.18 (t; 18H, J=6.0Hz, 2-(CH3) 3);
Embodiment 19:((2-(2-oxygen base-4-alkynes rosickyite yl pyrimidines-dihydro-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen is for the synthesis of dimethylene propylene carbonate (6d2)
According to the synthetic method of compound 6b2, the 5d of 1mmol, 4mmol triethylamine and 4mmol chloromethyl propylene carbonate are obtained by reacting the faint yellow paste 6d2 of about 0.16g in dry DMF, and productive rate is 29%, EI-MS:m/z537.4(M+H
+), 554.5(M+Na
+);
Product spectrum analytical data:
1HNMR (600MHz, DMSO-d6) δ ppm:8.0 (d, 1H, J=7.2Hz, CH); 6.39 (d, 1H, J=7.2Hz, CH), 5.34 (d; 4H, J=2.4Hz, 2CH2-O-P), 4,78-4.73 (m; 2H, 2CH-(CH3) 2), 3.97-3.93 (m, 4H; ≡ C-CH2 and N-CH2-CH2), 3.67 (t, 2H, J=6.6Hz; CH2-CH2-O), 3.30-3.34 (m, 3H, O-CH2-P; 1H, ≡ C-H), 1.21 (d, 12H, J=6.6Hz, 2-(CH3) 2);
Embodiment 20:((2-(2-oxygen base-4-alkynes rosickyite yl pyrimidines-dihydro-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen is for the synthesis of dimethylene carbonic acid benzyl (6d3)
According to the synthetic method of compound 6b3, the 5d of 1mmol, 4mmol triethylamine and 4mmol chloromethyl benzylcarbonate are obtained by reacting the faint yellow paste 6d3 of about 0.15g in dry DMF, and productive rate is 25%, EI-MS:m/z611.6(M+H
+), 633.6(M+Na
+);
Product spectrum analytical data:
1HNMR (600MHz, DMSO-d6) δ ppm:7.85 (d, 1H, J=6.6Hz; CH), 7.40-7.37 (m, 10H2 × 5H-Ar), 6.45 (d; 1H, J=7.2Hz, CH), 5.35 (d; 4H, J=2.4Hz, 2CH2-O-P); 4.65 (s, 1H, 2CH2-Ar); 3.99-3.97 (m, 4H, ≡ C-CH2 and N-CH2-CH2); 3.83 (d, 2H, J=8.4Hz; O-CH2-P), 3.76 (t, 2H; J=6.6Hz, CH2-CH2-O), 3.18 (t; 1H, J=3.6Hz, ≡ C-H);
Embodiment 21:((2-(4-benzylthio-2-oxygen base-pyrimidine-dihydro-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen is for the synthesis of the two dimethyl propylene acid esters (6e1) of dimethylene-2,2-
According to the synthetic method of compound 6b1, the 5e of 1mmol, 4mmol triethylamine and 4mmol trimethylacetic acid methyl esters are obtained by reacting the faint yellow paste 6e1 of about 0.16g in dry DMF, and productive rate is 27%, EI-MS:m/z585.5(M+H
+);
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：7.84(d,1H,J=7.2Hz,CH),7.41-7.27(m,5H,5H-Ar),6.39(d,1H,J＝7.2Hz,CH),5.41(d,4H,J＝2.4Hz,P-O-CH2),4.40(s,2H,CH2-Ar),3.96(t,2H,J＝6.6Hz,N-CH2-CH2),3.74(t,2H,J＝6.6Hz,CH2-CH2-O),3.59(d,2H,J＝8.4Hz,O-CH2-P),1.22(s,18H,2-(CH3)3);
Embodiment 22:((2-(4-benzylthio-2-oxygen base-pyrimidine-dihydro-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen is for the synthesis of dimethylene propylene carbonate (6e2)
According to the synthetic method of compound 6b2, the 5e of 1mmol, 4mmol triethylamine and 4mmol chloromethyl propylene carbonate are obtained by reacting the faint yellow paste 6e2 of about 0.14g in dry DMF, and productive rate is 24%, EI-MS:m/z589.5(M+H
+);
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：7.85(d,1H,J＝7.2Hz,CH),7.41-7.26(m,5H,5H-Ar),6.38(d,1H,J＝7.2Hz,CH),5.28(d,4H,J＝2.4Hz,2P-O-CH2),4.56-4.52(m,2H,2CH-(CH3)2),4.39(s,2H,CH2-Ar),3.97(t,2H,J＝6.6Hz,N-CH2-CH2),3.74(t,2H,J＝6.6Hz,CH2-CH2-O),3.58(d,2H,J＝8.4Hz,O-CH2-P),1.17(d,12H,J＝6.0Hz,2-(CH3)2);
Embodiment 23:((2-(4-benzylthio-2-oxygen base-pyrimidine-dihydro-1(2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen is for the synthesis of dimethylene benzylcarbonate (6e3)
According to the synthetic method of compound 6b3, the 5e of 1mmol, 4mmol triethylamine and 4mmol chloromethyl benzylcarbonate are obtained by reacting the faint yellow paste 6e3 of about 0.22g in dry DMF, and productive rate is 32%, EI-MS:m/z685.6(M+H
+), 707.7(M+Na
+);
Product spectrum analytical data:
1HNMR(600MHz,DMSO-d6)δppm：7.85(d,1H,J＝7.2Hz,CH),7.41-7.25(m,15H,3×5H-Ar),6.38(d,1H,J＝7.2Hz,CH),5.35(d,4H,J＝2.4Hz,P-O-CH2),4.73(s,4H,2CH2-Ar),4.39(s,2H,CH2-Ar),3.97(t,2H,J＝6.6Hz,N-CH2-CH2),3.75(t,2H,J＝6.6Hz,CH2-CH2-O),3.58(d,2H,J＝8.4Hz,O-CH2-P);
Embodiment 24: Anti-HBV activity cytoactive shaker test
Human hepatitis B virus transgenosis liver cancer cell (HepG2.2.15) is inoculated in 96 orifice plates, adds the example pharmaceuticals of different concns next day, changed nutrient solution and the example pharmaceuticals with concentration at the 4th day, to be measured in the 8th day collection culture supernatant.The culture supernatant of 50 μ l is added in good stripe board respectively to bag, and add HBeAg and HBsAg enzyme mark binding substances (blank control wells does not add) of equivalent, abundant mixing, stick gummed paper, liquid in stripe board is discarded after hatching 30 minutes at putting 37 DEG C, and with washings washing, repeat 5 times, then pat dry.Finally first add nitrite ion A(50 μ l), then add developer B(50 μ l), add stop buffer (50 μ l) termination reaction after lucifuge hatches 15 minutes at fully mixing is placed in 37 DEG C, measure OD by microplate reader
450/630, and according to the half inhibiting rate (IC of OD value calculation sample to HBV antigen
50).In addition, add MTT to the cell in 96 orifice plates, add the reaction of MTT lysate after 4 hours and spend the night, next day surveys OD570 in microplate reader.Calculate example pharmaceuticals to the toxic action of HepG2.2.15 cell and the situation affecting Growth of Cells according to OD value, finally calculate the concentration (CC caused needed for half necrocytosis amount
50).
The calculating of selectivity index: SI=CC
50/ EC
50.
Carried out Anti-HBV effect screening to 22 compounds of above-mentioned synthesis, their activity and toxicity data are listed in table 2, and the ucleosides Anti-HBV drugs lamivudine of wherein clinical application and adefovir ester are as positive control.
The structural formula of table 1 (2-(2-oxygen base-4-thiopyrimidine) oxyethyl group) methyl-phosphorous acid ester derivative
Table 2:(2-(2-oxygen base-4-thiopyrimidine) oxyethyl group) anti-HBsAg of methyl-phosphorous acid ester derivative and HBeAg is active and toxicity data
As can be seen from Table 2, thiopyrimidine acyclic nucleoside phosphonate analogue major part all remains certain anti-hepatitis B virus activities, contrasts different R
1the IC of substituent thionucleoside analogue
50value, can find out, the activity of compound d series compound is better, its IC
50value is starkly lower than other R
1the thionucleoside analogue that substituting group is modified, even lower than the IC contrasting medicine lamivudine and adefovir ester
50value, shows good Hepatitis B Surface S antigenic activity (HBsAg), the wherein IC of compound 6d1
50be worth 12.01 μm of ol/L, than the IC contrasting medicine lamivudine and adefovir ester
50reduce about half.The lead compound that can be used as Anti-HBV activity is used.
Claims (6)
1. one kind (2-(2-oxygen base-4-thiopyrimidine) oxyethyl group) methyl-phosphorous acid ester derivative has the structure shown in formula I:
Wherein,
R
1for methyl, allyl group, propargyl or benzyl;
R
2for hydrogen, ethyl, trimethylacetic acid methyl, isopropoxy carbonyl oxygen methyl or benzyloxycarbonyloxy methyl.
2. compound as claimed in claim 1, it is characterized in that one of following compound:
1) (2-(4-methylthio group-4-sulfo--2 oxopyrimidin-1 (2H)-Ji) oxyethyl group) methyl-phosphorous acid diethyl ester (4b),
2) (2-(4-allyl sulfo--2-oxopyrimidin-1 (2H)-Ji) oxyethyl group) methyl-phosphorous acid diethyl ester (4c),
3) (2-(2-oxygen base-4-propargyl thiopyrimidine-1 (2H)-Ji) oxyethyl group) methyl-phosphorous acid diethyl ester (4d),
4) (2-(4-benzene first sulphur-2-oxopyrimidin-1 (2H)-Ji) oxyethyl group) methyl-phosphorous acid diethyl ester (4e),
5) (2-(4-methylthio group-2-oxopyrimidin-1 (2H)-Ji) oxyethyl group) methyl-phosphorous acid (5b),
6) (2-(4-allyl sulfenyl-2-oxopyrimidin-1 (2H)-Ji) oxyethyl group) methyl-phosphorous acid (5c),
7) (2-(4-propargyl sulfenyl-2-oxopyrimidin-1 (2H)-Ji) oxyethyl group) methyl-phosphorous acid (5d),
8) (2-(2-oxygen base-4-thiophenyl pyrimidine) oxyethyl group) methyl-phosphorous acid (5e),
9) (2-(4-methylthio group-2-oxopyrimidin) methyl) phosphoryl dioxygen for the two dimethyl propylene acid esters (6b1) of dimethylene-2,2-,
10) ((2-(4-methylthio group-2-oxopyrimidin-1 (2H)-Ji) methyl) phosphoryl dioxygen for dimethylene propylene carbonate (6b2),
11) ((2-(4-methylthio group-2-oxopyrimidin-1 (2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for dimethylene benzylcarbonate (6b3),
12) ((2-(4-allyl sulfenyl-2-oxopyrimidin-1 (2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for the two dimethyl propylene acid esters (6c1) of dimethylene-2,2-,
13) ((2-(4-allyl sulfenyl-2-oxopyrimidin-1 (2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for dimethylene propylene carbonate (6c2),
14) ((2-(4-allyl sulfenyl-2-oxopyrimidin-1 (2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for dimethylene benzylcarbonate (6c3),
15) 2-(2-oxygen base-(4-alkynes rosickyite yl pyrimidines-1 (2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for the two dimethyl propylene acid esters (6d1) of dimethylene-2,2-,
16) ((2-(2-oxygen base-4-alkynes rosickyite yl pyrimidines-1 (2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for dimethylene propylene carbonate (6d2),
17) ((2-(2-oxygen base-4-alkynes rosickyite yl pyrimidines-1 (2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for dimethylene carbonic acid benzyl (6d3),
18) ((2-(4-benzylthio-2-oxygen base-pyrimidine-1 (2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for the two dimethyl propylene acid esters (6e1) of dimethylene-2,2-,
19) ((2-(4-benzylthio-2-oxygen base-pyrimidine-1 (2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen for dimethylene propylene carbonate (6e2) or
20) ((2-(4-benzylthio-2-oxygen base-pyrimidine-1 (2H)-Ji) oxyethyl group) methyl) phosphoryl dioxygen is for dimethylene benzylcarbonate (6e3).
3. the preparation method of compound as claimed in claim 1, step is as follows:
Take uridylic as starting raw material, key intermediate diethyl (2-(2 is shortened into [(2-chloroethoxy) methyl] diethyl phosphonate under alkali effect, 4-dioxy-1,4-dihydro-1 (2H)-Ji) oxyethyl group) methyl esters 2a, form target compound 3a via thiophosphoric anhydride sulfo-; Compound 3a and methyl iodide, allyl bromide 98, propargyl bromide or bromotoluene condensation obtain target compound 4b, 4c, 4d or 4e, obtain target compound 5b, 5c, 5d or 5e via hydrolysis, then obtain target compound 6b1-6b3,6c1-6c3,6d1-6d3 or 6e1-6e3 respectively at trimethylacetic acid methyl esters, chloromethyl base propylene carbonate or the condensation of chloromethyl benzylcarbonate;
Synthetic route is as follows:
Reagent and condition: (i) salt of wormwood, sodium methylate, DMF; (ii) pyridine, thiophosphoric anhydride, 120 DEG C, 2h; (iv) acetone, salt of wormwood, backflow; (v) acetonitrile, bromotrimethylsilane; (vi) DMF, triethylamine.
4. preparation method as claimed in claim 3, is characterized in that step is as follows:
(1) synthesis of intermediate (2-(2,4-dioxy-Isosorbide-5-Nitrae-dihydro-1 (2H)-Ji) oxyethyl group) methyl acid phosphate diethyl ester (2a)
By 10g uridylic and the anhydrous N of 200mL, N-dimethylformamide joins in 500mL round-bottomed flask, 120 DEG C of reflux 1 are little all dissolves up to uridylic, add 6.2g salt of wormwood again, 0.48g sodium methylate and 20.5g [(2-chloroethoxy) methyl] diethyl phosphonate, react 8h under 120 DEG C of conditions; TLC detection reaction is complete, is evaporated to pasty state, adds about 150mL methylene dichloride, elimination insolubles, and filtrate concentrates, and upper silica gel column chromatography is separated, and moving phase selects volume ratio 30:1 methylene dichloride-methyl alcohol, and obtain colorless oil 2a, productive rate is 34%;
(2) synthesis of (2-(2-oxo-4-sulfo--3,4-dihydro-pyrimidin-1 (2H)-Ji) oxyethyl group) methyl-phosphorous acid diethyl ester (3a)
The 2a of 5g is dissolved in 50mL pyridine, is heated to 120 DEG C, react 1 hour, TLC detection reaction reduces pressure steam solvent completely, adds a large amount of frozen water in residue, filters insolubles, solution with ethyl acetate extraction 3 ~ 4 times, organic layer adds anhydrous sodium sulfate drying, filters solvent evaporated and obtains crude product; Crude product silica gel column chromatography is separated, and moving phase is volume ratio 3:1 ethyl acetate-methylene dichloride, turns the methylene dichloride of volume ratio 20:1: methyl alcohol, obtains yellow oil 3a;
(3) synthesis of (2-(-4-sulfo--2 oxopyrimidin-dihydro-1 (2H)-Ji) oxyethyl group) methyl-phosphorous acid diethyl ester (4b-4e)
6mmol2a is dissolved in 15mL acetone, adds 3mmol salt of wormwood and 6mmol methyl iodide, bromopropylene, propargyl bromide or bromotoluene, 80 DEG C of back flow reaction 2 hours, TLC detection reaction is complete, and decompression steams solvent, is separated obtains pale yellow oil 4b respectively with silica gel column chromatography, 4c, 4d or 4e;
(4) synthesis of (2-(4-(methylthio group/allyl sulfenyl/alkynes rosickyite base/thiophenyl)-2-oxopyrimidin-dihydro-1 (2H)-Ji) oxyethyl group) methyl-phosphorous acid (5b, 5c, 5d or 5e)
1g4b, 4c, 4d or 4e and bromotrimethylsilane react 18 hours in acetonitrile, obtain yellow paste 5b respectively, 5c, 5d or 5e;
(5) synthesis of (2-(4-methylthio group-2-oxopyrimidin-dihydro-1 (2H)-Ji) oxyethyl group) methylphosphonate (6b1,6b2 or 6b3)
The 5b of 1mmol is dissolved in the anhydrous DMF of 5mL, in stirring at room temperature half hour, adds 4mmol triethylamine, continues stirring 1 at room temperature little muddy up to solution; Then 4mmol trimethylacetic acid methyl esters is added, chloromethyl propylene carbonate or chloromethyl benzylcarbonate are warming up to 60 DEG C of reactions 7 hours, TLC detection reaction is complete, add 15mL ethyl acetate to stir 1 hour at 0 DEG C, filter, filtrate evaporate to dryness, be separated with silica gel column chromatography, moving phase is volume ratio 15:1 methylene dichloride-methyl alcohol, obtains faint yellow paste 6b1,6b2 or 6b3;
(6) synthesis of (2-(4-allyl sulfenyl-2-oxopyrimidin-dihydro-1 (2H)-Ji) oxyethyl group) methylphosphonate (6c1,6c2 or 6c3)
The 5c of 1mmol is dissolved in the anhydrous DMF of 5mL, in stirring at room temperature half hour, adds 4mmol triethylamine, continues stirring 1 at room temperature little muddy up to solution; Then 4mmol trimethylacetic acid methyl esters is added, chloromethyl propylene carbonate or chloromethyl benzylcarbonate, be warming up to 60 DEG C of reactions 7 hours, TLC detection reaction is complete, adds 15mL ethyl acetate and stirs 1 hour at 0 DEG C, filter, filtrate evaporate to dryness, be separated with silica gel column chromatography, moving phase is volume ratio 15:1 methylene dichloride-methyl alcohol, obtain faint yellow paste 6c1,6c2 or 6c3;
(7) synthesis of (2-(4-alkynes rosickyite base-2-oxopyrimidin-dihydro-1 (2H)-Ji) oxyethyl group) methylphosphonate (6d1,6d2 or 6d3)
The 5d of 1mmol is dissolved in the anhydrous DMF of 5mL, in stirring at room temperature half hour, adds 4mmol triethylamine, continues stirring 1 at room temperature little muddy up to solution; Then 4mmol trimethylacetic acid methyl esters is added, chloromethyl propylene carbonate or chloromethyl benzylcarbonate, be warming up to 60 DEG C of reactions 7 hours, TLC detection reaction is complete, adds 15mL ethyl acetate and stirs 1 hour at 0 DEG C, filter, filtrate evaporate to dryness, be separated with silica gel column chromatography, moving phase is the methylene dichloride-methyl alcohol of volume ratio 15:1, obtains faint yellow paste 6d1,6d2 or 6d3;
(8) synthesis of (2-(4-thiophenyl-2-oxopyrimidin-dihydro-1 (2H)-Ji) oxyethyl group) methylphosphonate (6e1,6e2 or 6e3)
The 5e of 1mmol is dissolved in the anhydrous DMF of 5mL, in stirring at room temperature half hour, adds 4mmol triethylamine, continues stirring 1 at room temperature little muddy up to solution; Then 4mmol trimethylacetic acid methyl esters is added, chloromethyl propylene carbonate or chloromethyl benzylcarbonate are warming up to 60 DEG C of reactions 7 hours, TLC detection reaction is complete, add 15mL ethyl acetate to stir 1 hour at 0 DEG C, filter, filtrate evaporate to dryness, be separated with silica gel column chromatography, moving phase is methylene dichloride: methyl alcohol 15:1, obtains faint yellow paste 6e1,6e2 or 6e3.
5. compound as claimed in claim 1 or 2 is preparing the application in Anti-HBV drugs.
6. an Anti-HBV drugs composition, is characterized in that comprising the compound described in claim 1 or 2 and one or more pharmaceutically acceptable carriers or vehicle.
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