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CN1337401A - Thiophosphoryl amino acid ester compound and its prepn process - Google Patents

Thiophosphoryl amino acid ester compound and its prepn process Download PDF

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CN1337401A
CN1337401A CN 01130784 CN01130784A CN1337401A CN 1337401 A CN1337401 A CN 1337401A CN 01130784 CN01130784 CN 01130784 CN 01130784 A CN01130784 A CN 01130784A CN 1337401 A CN1337401 A CN 1337401A
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isopropylidene
nmr
compound
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1mmol
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CN1159332C (en
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赵玉芬
苗志伟
付华
成昌梅
韩波
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Tsinghua University
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Tsinghua University
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Abstract

本发明涉及一种硫代磷酰氨基酸酯化合物,结构式为:其制备方法为首先将原料(异丙氧基取代)二氯硫磷溶于四氢呋喃中,向上述溶液中加入氨基酸甲酯盐酸盐,再滴加缚酸剂,将2′,3′－O－异亚丙基核苷溶液加入上述溶液中,再滴加缚酸剂,反应完成后,过滤,旋转蒸馏,,柱层析分离,即得硫代磷酰氨基酸酯化合物。

The invention relates to a thiophosphoryl amino acid ester compound, the structural formula of which is as follows: the preparation method is firstly dissolving the raw material (isopropoxy substituted) thionyl chloride in tetrahydrofuran, and adding amino acid methyl ester hydrochloride to the above solution , then add the acid-binding agent dropwise, add the 2′,3′-O-isopropylidene nucleoside solution into the above solution, then add the acid-binding agent dropwise, after the reaction is completed, filter, rotate and distill, and separate by column chromatography , That is, phosphorothioate amino acid ester compound.

Description

A kind of thiophosphoryl amino acid ester compound and preparation method thereof
The present invention relates to a kind of thiophosphoryl amino acid ester compound and preparation method thereof, relate in particular to 2 ', 3 '-O-isopropylidene nucleosides 5 '-preparation method of (isopropoxy replacement) thiophosphoryl amino acid ester compound, this compounds has good biological activity and antiviral, antitumor, anti-ly likes now pharmaceutical activity such as virus, can develop in clinical application that a kind of ucleosides is antiviral, antitumor, anti-likes viral now prodrug (prodrugs).
Thiophosphoric acid contains the structure that a sulphur replaces oxygen on phosphorus atom, though this structure is still keeping original electric charge, the character that is keeping highly water-soluble, but other physicochemical properties of the dna oligo of thiophosphoric acid (S-oligos) and biology attribute all have very big difference with natural phosphodiester prototype.One of marked difference is that thiophosphoric acid has resistivity to nuclease.Because the phosphodiester class antisense oligonucleotide that adds has been fallen in the existence of exonuclease in the cell, very fast digestion, make its forfeiture action function.By synthetic thiophosphoric acid compounds with opposing nuclease, be used for the treatment of purpose for using the synthetic dna oligo from now on, opened up the road of a hope beyond doubt.
The objective of the invention is to propose a kind of thiophosphoryl amino acid ester compound and preparation method thereof, specifically, be 2 ', 3 '-O-isopropylidene nucleosides 5 '-preparation method of (isopropoxy replacement) thiophosphoryl amino acid ester compound, to be used to develop antiviral, antitumor, the anti-active medicine of virus now of liking.
2 of the present invention's proposition ', 3 '-O-isopropylidene nucleosides 5 '-(isopropoxy replacement) thiophosphoryl amino acid ester compound, its structural formula is: R is H in the said structure formula, CH 3, C 6H 5CH 2, (CH 3) 2CH 2, (CH 3) 2CHCH 2B is a VITAMIN B4, guanine, cytosine(Cyt), uridylic.
The preparation method of above-claimed cpd comprises the steps:
1) under nitrogen protection, cryosel is bathed under the condition that is cooled to-4～-8 ℃, and raw material (isopropoxy replacement) two compd 22/190s are dissolved in the dry tetrahydrofuran (THF) of crossing (THF), is mixed with the solution that concentration is 0.8～1.0mol/L.
2) add amino acid methyl ester hydrochloride with the amount of raw material same substance in above-mentioned solution, slowly drip the acid binding agent triethylamine after stirring, the add-on of acid binding agent is 2 times of amount of raw material.
3) preparation 2 ', 3 '-O-isopropylidene nucleosides solution for standby:
Get nucleosides under the room temperature and be suspended in the solution that is made into 0.5～1.0mol/L in the exsiccant solvent, while stirring to wherein adding and the tosic acid of amount of substances such as nucleosides and the triethyl orthoformate of four times of amount of substances, after system becomes clarification, neutralize with big water gaging with strong aqua and to make PH=7～8, then above-mentioned solution decompression is concentrated, in refrigerator standing over night can obtain product 2 ', 3 '-O-isopropylidene nucleosides.With make 2 ', 3 '-be dissolved in the solution that is mixed with 0.5～1.0mol/L in the solvent after the O-isopropylidene nucleosides suction filtration drying.
4) follow the tracks of above-mentioned second reaction process that goes on foot, after treating that the raw material total overall reaction finishes, will with 2 of above-mentioned the 3rd step preparation of the amount of raw material same substance ', 3 '-O-isopropylidene nucleosides solution slowly splashes in the solution in second step, and the back that stirs continue to drip the acid binding agent triethylamine with the amount of raw material same substance.
5) after monitoring reaction is finished, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.Recycle silicon glue post carries out column chromatography for separation, can obtain 2 ', 3 '-O-isopropylidene nucleosides 5 '-(isopropoxy replacement) thiophosphoryl amino acid ester compound.
With 2 of method of the present invention preparation ', 3 '-O-isopropylidene nucleosides 5 '-(isopropoxy replacement) thiophosphoryl amino acid methyl ester, be a class brand-new have an anti-active compound of virus drugs now of liking, by synthetic different types of nucleosides-amino acid conjugate, and carry out activity experiment, tentatively obtained satisfied result, liked now the virus drugs achievement in research that provides the foundation for further development and development of new resist.
Below introduce embodiments of the invention:
Embodiment 1: preparation 2 ', '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl glycine methyl ester compound, wherein R is H, B is a VITAMIN B4.
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the glycine methyl ester hydrochloride of 1mmol (0.125g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) preparation 2 ', 3 '-O-isopropylidene adenosine is standby:
Getting 1mol (267g) adenosine under the room temperature is suspended in the acetone of 2L, stir down to wherein adding 1.1mol (210g) tosic acid and 4mol (640ml) triethyl orthoformate, system becomes clarification after about 1 hour, neutralize with 100ml strong aqua and 5.9L water, then above-mentioned solution decompression is concentrated into 1.5L, in refrigerator standing over night can obtain product 2 ', 3 '-O-isopropylidene adenosine 266g (87%).
4) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene adenosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) after the reaction of monitoring above-mentioned second step with NMR is finished, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.Carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl glycine methyl ester, productive rate is 58.4%.
Spectral data is as follows: 31P NMR (DMSO-d 6, δ: ppm, J:Hz): δ 60.38,61.02; 1H NMR (500MHz, DMSO-d 6): δ 9.26,9.22 (bs, 1H, NH), 8.43 (1H, s, H-2), 8.23 (1H, s, H-8), 7.40 (2H, s, NH 2), 6.44 (1H, m, H-1 '), 4,50 (2H, m, H-2 ', 3 '), 3.97 (1H, m, H-4 '), 3.88 (3H, s, OCH 3), 3.64 (2H, m, H-5 '), 3.57 (2H, m, H-α), 1.54 (3H, s, CH 3), 1.33 (3H, s, CH 3); 13C NMR (500MHz, DMSO-d 6): δ 173.24 (COOMe), 163.70 (C-2), 150.43 (C-4), 140.76 (C-6), 117.02,116.96 (＞CMe 2), 101.76 (C-1 '), 87.74 (C-5), 84.83 (C-2 '), 73.54 (C-3 '), 69.80 (C-4 '), 60.87 (C-5 '), 54.78 (OCH 3), 45.86 (C-α), 26.40 (CH 3), 26.86 (CH 3); ESI-MS (pos.): m/z 476 (M+H) +ESI-MS (neg.): m/z 474 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
Like virus=Human immunodeficiency virus now
MT-4＝Human?leukenia?T?cell
CEM＝Human?lymphoblastoid?T?cell
ED 50=antiviral activity index
CD 50=cytotoxicity index
ED 50?CEM-TK- 5×10 -3M (CD 50?7×10 -6M)
CEM-SS 4×10 -3M (CD 60?8×10 -6M)
MT4 2×10 -3M (CD 60?8×10 -6M)
Embodiment 2: preparation 2 ', 3 '-O-isopropylidene adenosine 5 '-thiophosphoryl alanine methyl ester compound, wherein R is CH 3, B is a VITAMIN B4.
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the alanine methyl ester hydrochloride of 1mmol (0.14g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene adenosine synthetic with above-mentioned embodiment (1).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene adenosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl alanine methyl ester, productive rate is 62.4%.
Spectral data is as follows: 31P NMR (DMSO-d 6, δ: ppm, J:Hz): δ 57.86,57.03; 1H NMR (500MHz, DMSO-d 6): δ 9.44,9.40 (bs, 1H, NH), 8.89 (1H, s, H-2), 8.67 (1H, s, H-8), 7.67 (2H, s, NH 2), 6.59 (1H, m, H-1 '), 4,78 (2H, m, H-2 ', 3 '), 4.22 (1H, m, H-4 '), 3.98 (3H, s, OCH 3), 3.85 (2H, m, H-5 '), 3.67 (2H, m, H-α), 1.61 (3H, s, CH 3), 1.42 (3H, s, CH 3), 1.31,1.30 (3H, d, 3J=6, β-CH 3); 13C NMR (500MHz, DMSO-d 6): δ 180.24 (COOMe), 170.70 (C-2), 156.43 (C-4), 149.76 (C-6), 117.02,116.96 (＞CMe 2), 108.23 (C-1 '), 89.66 (C-5), 85.87 (C-2 '), 76.23 (C-3 '), 69.66 (C-4 '), 61.23 (C-5 '), 54.78 (OCH 3), 50.73 (C-α), 48.66 (C-β), 26.40 (CH 3), 26.86 (CH 3); ESI-MS (pos.): m/z 487 (M+H) +ESI-MS (neg.): m/z 489 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 4×10 -3M (CD 50?2×10 -6M)
CEM-SS 6×10 -3M (CD 50?9×10 -6M)
MT4 8×10 -3M (CD 50?6×10 -6M)
Embodiment 3: preparation 2 ', 3 '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl phenylalanine methyl ester compound, wherein R is C 6H 5CH 2, B is a VITAMIN B4.
The structural formula of compound:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the phenylalanine methyl ester hydrochloride of 1mmol (0.22g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene adenosine synthetic with above-mentioned embodiment (1).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene adenosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl phenylalanine methyl ester, productive rate is 58.7%.
Spectral data is as follows: 31P NMR (DMSO-d 6, δ: ppm, J:Hz): δ 58.64,58.33; 1H NMR (500MHz, DMSO-d 6): δ 9.21,9.15 (bs, 1H, NH), 8.67 (1H, s, H-2), 8.58 (1H, s, H-8), 7.88 (2H, s, NH 2), 7.15-7.35 (5H, m, Ph), 6.59 (1H, m, H-1 '), 4,55 (2H, m, H-2 ', 3 '), 4.37 (1H, m, H-4 '), 3.79 (3H, s, OCH 3), 3.66 (2H, m, H-5 '), 3.48 (2H, m, H-α), 2.32 (2H, m, H-β), 1.61 (3H, s, CH 3), 1.49 (3H, s, CH 3); 13C NMR (500MHz, DMSO-d 6): δ 174.24 (COOMe), 170.60 (C-2), 159.43 (C-4), 148.36 (Ph-jpso), 117.02,116.96 (＞CMe 2), 136.11 (C-6), 135.12 (Ph-para), 130.42 (Ph-ortho), 119.91 (Ph-meta), (108.23 C-1 '), 89.65 (C-5), 87.87 (C-2 '), 76.23 (C-3 '), (72.66 C-4 '), 61.23 (C-5 '), 56.82 (C-β), 56.78 (OCH 3), 52.73 (C-α), 26.40 (CH 3), 23.80 (CH 3); ESI-MS (pos.): m/z 565 (M+H) +ESI-MS (neg.): m/z 563 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 8×10 -3M (CD 50?4×10 -6M)
CEM-SS 9×10 -3M (CD 50?7×10 -6M)
MT4 6×10 -3M (CD 50?8×10 -6M)
Embodiment 4: preparation 2 ', 3 '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester compound, wherein R is (CH 3) 2CHCH 2, B is a VITAMIN B4.The structural formula of compound is:
Figure A0113078400071
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the valine methyl ester hydrochloride of 1mmol (0.17g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene adenosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
4) 2 ', 3 '-O-isopropylidene adenosine synthetic with above-mentioned embodiment (1).
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester, productive rate is 55.2%.
Spectral data is as follows: 31P NMR (DMSO-d 6, δ: ppm, J:Hz): δ 56.32,56.14; 1H NMR (500MHz, DMSO-d 6): δ 9.44,9.40 (bs, 1H, NH), 8.67 (1H, s, H-2), 8.58 (1H, s, H-8), 7.54 (2H, s, NH 2), 6.59 (1H, m, H-1 '), 4,78 (2H, m, H-2 ', 3 '), 4.22 (1H, m, H-4 '), 3.98 (3H, s, OCH 3), 3.85 (2H, m, H-5 '), 3.67 (1H, m, H-α), 3.45 (1H, m, H-β), 1.61 (3H, s, CH 3), 1.42 (3H, s, CH 3), 1.31 (3H, d, 3J=6, CH 3), 1.25 (3H, d, 3J=6, CH 3); 13C NMR (500MHz, DMSO-d 6): δ 172.24 (COOMe), 165.70 (C-2), 159.43 (C-4), 145.76 (C-6), 117.02,116.96 (＞CMe 2), 108.23 (C-1 '), 89.66 (C-5), 85.87 (C-2 '), 76.23 (C-3 '), 69.66 (C-4 '), 61.23 (C-5 '), 54.78 (OCH 3), 50.73 (C-α), 48.66 (C-β), 26.40 (CH 3), 26.86 (CH 3); ESI-MS (pos.): m/z 518 (M+H) +ESI-MS (neg.): m/z 516 (M-H) -. the anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 5×10 -3M (CD 50?2×10 -6M)
CEM-SS 6×10 -3M (CD 50?5×10 -6M)
MT4 8×10 -3M (CD 50?6×10 -6M)
Embodiment 5: preparation 2 ', 3 '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl leucine methyl compound, wherein R is (CH 3) 2CHCH 2, B is a VITAMIN B4.
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the leucine methyl ester hydrochloride of 1mmol (0.18g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene adenosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
4) 2 ', 3 '-O-isopropylidene adenosine synthetic with above-mentioned embodiment (1).
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene adenosine 5 '-thiophosphoryl leucine methyl esters, productive rate is 53.2%.
Spectral data is as follows: 31P NMR (DMSO-d 6, δ: ppm, J:Hz): δ 54.32,54.14; 1H NMR (500MHz, DMSO-d 6): δ 9.31,9.27 (bs, 1H, NH), 8.67 (1H, s, H-2), 8.58 (1H, s, H-8), 7.54 (2H, s, NH 2), 6.59 (1H, m, H-1 '), 4,78 (2H, m, H-2 ', 3 '), 4.22 (1H, m, H-4 '), 3.98 (3H, s, OCH 3), 3.85 (2H, m, H-5 '), 3.67 (1H, m, H-α), 3.45 (1H, m, H-β), 3.21 (1H, m, H-γ), 1.61 (3H, s, CH 3), 1.42 (3H, s, CH 3), 1.31 (3H, d, 3J=6, CH 3), 1.25 (3H, d, 3J=6, CH 3); 13C NMR (500MHz, DMSO-d 6): δ 172.24 (COOMe), 165.70 (C-2), 159.43 (C-4), 145.76 (C-6), 116.02,115.96 (＞CMe 2), 109.23 (C-1 '), 89.66 (C-5), 85.87 (C-2 '), 76.23 (C-3 '), 69.66 (C-4 '), 61.23 (C-5 '), 54.78 (OCH 3), 50.73 (C-α), 48.66 (C-β), 46.35 (C-γ), 23.40 (CH 3), 22.86 (CH 3); ESI-MS (pos.): m/z 531 (M+H) +ESI-MS (neg.): m/z 529 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 5×10 -3M (CD 50?2×10 -6M)
CEM-SS 2×10 -3M (CD 50?5×10 -6M)
MT4 5×10 -3M (CD 50?9×10 -6M)
Embodiment 6: preparation 2 ', 3 '-O-isopropylidene guanosine 5 '-thiophosphoryl glycine methyl ester compound, wherein R is H, B is a guanine.The structural formula of compound is: The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the glycine methyl ester hydrochloride of 1mmol (0.125g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) preparation 2 ', 3 '-O-isopropylidene guanosine is standby
Getting 1mol (286g) guanosine under the room temperature is suspended in the acetone of 2L, stir down to wherein adding 1.1mol (210g) tosic acid and 4mol (640ml) triethyl orthoformate, system becomes clarification after about 1 hour, neutralize with 100ml strong aqua and 5.9L water, this moment product 2 ', 3 '-O-isopropylidene guanosine slowly separates out, standing over night under the room temperature, after the filtration again with ice-water mixture thorough washing, obtain after the drying product 2 ', 3 '-O-isopropylidene guanosine 240g (74%).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.323g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene guanosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) dichloro thiophosphoryl glycine methyl ester, productive rate is 58.4%.
Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 56.68,56.39; 1H NMR (500MHz, D 2O): δ 8.18 (1H, s, H-8), 5.82,5.81 (1H, d, 3J=6.0, H-1 '), 4,89 (1H, m, H-2 '), 4.23 (1H, m, H-3 '), 3.23 (1H, m, H-4 '), 3.54 (3H, s, OCH 3), 3.65 (2H, m, H-5 '), 3.54 (2H, m, H-α), 1.66,1.65 (3H, d, 3J=6.0, β-CH 3), 1.35 (3H, s, CH 3), 1.33 (3H, s, CH 3); 13C NMR (500MHz, D 2O): δ 172.31 (COOMe), 165.81 (C-2), 158.67 (C-6), 152.34 (C-4), 139.66 (C-8), 121.11 (＞CMe 2), 118.71 (C-5), 89.42 (C-4 '), 86.25 (C-1 '), 73.55 (C-3 '), 70.82 (C-2 '), 61.45 (C-5 '), 54.62 (OCH 3), 50.62 (C-β), 45.89 (C-α), 28.66 (CH 3), 26.1 5 (CH 3); ESI-MS (pos.): m/z 491 (M+H) +ESI-MS (neg.): m/z 489 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 8×10 -3M (CD 50?6×10 -6M)
CEM-SS 3×10 -3M (CD 50?4×10 -6M)
MT4 6×10 -3M (CD 50?3×10 -6M)
Embodiment 7:2 ', 3 '-O-isopropylidene guanosine 5 '-preparation of (isopropoxy replacement) thiophosphoryl alanine methyl ester compound, wherein R is CH 3, B is a guanine.
The structural formula of compound is:
Figure A0113078400091
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the alanine methyl ester hydrochloride of 1mmol (0.140g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene guanosine synthetic with above-mentioned embodiment (6).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene guanosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
4) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) thiophosphoryl alanine methyl ester, productive rate is 52.3%.
Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 55.88,55.37; 1H NMR (500MHz, D 2O): δ 8.07 (1H, s, H-8), 5.82,5.81 (1H, d, 3J=6.0, H-1 '), 4,51 (1H, m, H-2 '), 4.18 (1H, m, H-3 '), 3.94 (1H, m, H-4 '), 3.86 (3H, s, OCH 3), 3.68 (2H, m, H-5 '), 3.57 (2H, m, H-α), 1.35 (3H, s, CH 3), 1.33 (3H, s, CH 3); 13C NMR (500MHz, D 2O): δ 178.31 (COOMe), 156.81 (C-2), 153.67 (C-6), 151.34 (C-4), 135.66 (C-8), 120.11 (＞CMe 2), 116.71 (C-5), 86.42 (C-4 '), 85.25 (C-1 '), 73.74 (C-3 '), 70.42 (C-2 '), 61.45 (C-5 '), 54.62 (OCH 3), 45.89 (C-α), 23.66 (CH 3), 23.15 (CH 3); ESI-MS (pos.): m/z 506 (M+H) +ESI-MS (neg.): m/z 504 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 6×10 -3M (CD 50?7×10 -5M)
CEM-SS 6×10 -3M (CD 50?9×10 -6M)
MT4 2×10 -4M (CD 50?7×10 -5M)
Embodiment 8: preparation 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) thiophosphoryl phenylalanine methyl ester compound, wherein R is C 6H 5CH 2, B is a guanine.
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the phenylalanine methyl ester hydrochloride of 1mmol (0.21g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene guanosine synthetic with above-mentioned embodiment (6).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene guanosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) thiophosphoryl phenylalanine methyl ester, productive rate is 50.3%.
Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 57.18,56.89; 1H NMR (500MHz, D 2O): δ 8.07 (1H, s, H-8), 7.25-7.41 (5H, m, Ph), 5.79,5.78 (1H, d, 3J=6.0, H-1 '), 4,41 (1H, m, H-2 '), 4.18 (1H, m, H-3 '), 3.94 (1H, m, H-4 '), 3.86 (3H, s, OCH 3), 3.68 (2H, m, H-5 '), 3.57 (2H, m, H-α), 2.27 (2H, m, H-β), 1.21 (3H, s, CH 3), 1.17 (3H, s, CH 3); 13C NMR (500MHz, D 2O): δ 180.31 (COOMe), 166.81 (C-2), 161.67 (C-6), 158.34 (C-4), 151.26 (Ph-jpso), 135.66 (C-8), 130.07 (Ph-para), 128.23 (Ph-ortho), 118.45 (Ph-meta), 108.11 (＞CMe 2), 96.71 (C-5), 86.42 (C-4 '), 85.25 (C-1 '), 73.74 (C-3 '), 70.42 (C-2 '), 61.45 (C-5 '), 57.22 (C-β), 54.62 (OCH 3), 45.89 (C-α), 23.66 (CH 3), 23.15 (CH 3); ESI-MS (pos.): m/z 581 (M+H) +ESI-MS (neg.): m/z 579 (M-H) -. the anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 6×10 -3M (CD 50?7×10 -5M)
CEM-SS 6×10 -3M (CD 50?9×10 -6M)
MT4 2×10 -4M (CD 50?7×10 -5M)
Embodiment 9: preparation 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester compound, wherein R is (CH 3) 2CH 2, B is a guanine.
The structural formula of compound is:
Figure A0113078400111
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the valine methyl ester hydrochloride of 1mmol (0.170g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene guanosine synthetic with above-mentioned embodiment (6).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene guanosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester, productive rate is 56.2%.
Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 58.86,58.27; 1H NMR (500MHz, D 2O): δ 9.07 (1H, s, H-8), 6.82,6.81 (1H, d, 3J=6.0, H-1 '), 5,48 (1H, m, H-2 '), 4.95 (1H, m, H-3 '), 4.36 (1H, m, H-4 '), 3.86 (3H, s, OCH 3), 3.68 (2H, m, H-5 '), 3.57 (2H, m, H-α), 3.45 (1H, m, H-β), 1.35 (3H, s, CH 3), 1.31 (3H, s, CH 3), 1.16,1.15 (3H, d, 3J=6.0, CH 3), 1.06,1.05 (3H, d, 3J=6.0, CH 3); 13C NMR (500MHz, D 2O): δ 178.31 (COOMe), 156.81 (C-2), 153.67 (C-6), 151.34 (C-4), 135.66 (C-8), 120.11 (＞CMe 2), 116.71 (C-5), 86.42 (C-4 '), 85.25 (C-1 '), 73.74 (C-3 '), 70.42 (C-2 '), 61.45 (C-5 '), 54.62 (OCH 3), 45.89 (C-α), 44.24 (C-β), 23.66 (CH 3), 23.15 (CH 3), 2 1.26 (CH 3), 20.05 (CH 3); ESI-MS (pos.): m/z 533 (M+H) +ESI-MS (neg.): m/z 531 (M-H) -. the anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 5×10 -3M (CD 50?6×10 -5M)
CEM-SS 7×10 -2M (CD 50?9×10 -5M)
MT 44×10 -4M (CD 50?8×10 -5M)
Embodiment 10: preparation 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) thiophosphoryl leucine methyl compound, wherein R is (CH 3) 2CHCH 2, B is a guanine.
The structural formula of compound is:
Figure A0113078400121
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the leucine methyl ester hydrochloride of 1mmol (0.180g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene guanosine synthetic with above-mentioned embodiment (6).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene guanosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) thiophosphoryl leucine methyl esters, productive rate is 50.2%.
Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 55.86,55.27; 1H NMR (500MHz, D 2O): δ 9.07 (1H, s, H-8), 6.82,6.81 (1H, d, 3J=6.0, H-1 '), 5,48 (1H, m, H-2 '), 4.95 (1H, m, H-3 '), 4.36 (1H, m, H-4 '), 3.86 (3H, s, OCH 3), 3.68 (2H, m, H-5 '), 3.57 (2H, m, H-α), 3.45 (1H, m, H-β), 3.22 (1H, m, H-γ), 1.35 (3H, s, CH 3), 1.31 (3H, s, CH 3), 1.16,1.15 (3H, d, 3J=6.0, CH 3), 1.06,1.05 (3H, d, 3J=6.0, CH 3); 13C NMR (500MHz, D 2O): δ 178.31 (COOMe), 166.81 (C-2), 159.67 (C-6), 156.34 (C-4), 134.66 (C-8), 17.11 (＞CMe 2), 116.71 (C-5), 86.42 (C-4 '), 85.25 (C-1 '), 73.74 (C-3 '), 70.42 (C-2 '), 61.45 (C-5 '), 54.62 (OCH 3), 45.89 (C-α), 44.24 (C-β), 41.24 (C-γ), 21.26 (CH 3), 20.05 (CH 3); ESI-MS (pos.): m/z 548 (M+H) +ESI-MS (neg.): m/z 546 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 6×10 -3M (CD 50?6×10 -5M)
CEM-SS 8×10 -3M (CD 50?8×10 -6M)
MT4 8×10 -4M (CD 50?2×10 -5M)
Embodiment 11: preparation 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl glycine methyl ester compound, wherein R is H, B is a cytosine(Cyt).
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the glycine methyl ester hydrochloride of 1mmol (0.125g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) preparation 2 ', 3 '-O-isopropylidene cytidine is standby
Get 1mol (244g) guanosine under the room temperature and be suspended in the acetone of 2L, stir down to wherein adding 1.1mol (210g) tosic acid and 4mol (640ml) triethyl orthoformate, system change clarification after about 1 hour, adding 1mol (84g) NaHCO 3Neutralize, continue to stir 15 minutes, filter the back solids washed with acetone, with obtain behind filtrate and the washings evaporate to dryness product 2 ', 3 '-O-isopropylidene cytidine slowly separates out, standing over night under the room temperature, after the filtration again with ice-water mixture thorough washing, obtain after the drying product 2 ', 3 '-O-isopropylidene cytidine 263g (92%).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.282g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene cytidine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl glycine methyl ester, productive rate is 60.4%.Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 57.68,56.26; 1H NMR (500MHz, D 2O): δ 7.90,7.89 (1H, d, 3J=5.5, H-6), 6.24 (1H, m, H-1 '), 5.84,5.83 (1H, d, 3J=6, H-5), 4.28 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.71 (3H, s, OCH 3), 3.65 (2H, m, H-5 '), 2.69 (2H, m, H-α), 2.1 5 (1H, H-2 '), 1.1 5 (3H, s, CH 3), 1.07 (3H, s, CH 3); 13C NMR (500MHz, D 2O): δ 177.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 140.34 (C-6), 121.11 (＞CMe 2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH 3), 45.89 (C-α), 39.82 (C-2 '), 28.66 (CH 3), 26.15 (CH 3); ESI-MS (pos.): m/z 451 (M+H) +ESI-MS (neg.): m/z 449 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 8×10 -4M (CD 50?6×10 -5M)
CEM-SS 6×10 -3M (CD 50?4×10 -5M)
MT4 6×10 -4M (CD 50?3×10 -6M)
Embodiment 12: preparation 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl alanine methyl ester compound, wherein R is CH 3, B is a cytosine(Cyt).
The structural formula of compound is:
Figure A0113078400131
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and is cooled to-4～-8 ℃, with (isopropoxy replacement) dichloro of 1mmol (0.17g)
Sulphur phosphorus is dissolved in the dry tetrahydrofuran (THF) of crossing (THF), is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the alanine methyl ester hydrochloride of 1mmol (0.14g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene cytidine synthetic with above-mentioned embodiment 11.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.282g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene cytidine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl alanine methyl ester, productive rate is 62.4%.
Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 55.66,55.31; 1H NMR (500MHz, D 2O): δ 7.90,7.89 (1H, d, 3J=5.5, H-6), 6.88 (1H, m, H-1 '), 5.84,5.83 (1H, d, 3J=6, H-5), 4.68 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.71 (3H, s, OCH 3), 3.65 (2H, m, H-5 '), 2.89 (2H, m, H-α), 2.66 (1H, H-2 '), 1.45 (3H, d, 3J=6, H-β), 1.15 (3H, s, CH 3), 1.07 (3H, s, CH 3); 13C NMR (500MHz, D 2O): δ 185.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 145.34 (C-6), 121.11 (＞CMe 2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH 3), 50.86 (C-β), 45.89 (C-α), 39.82 (C-2 '), 27.66 (CH 3), 24.15 (CH 3); ESI-MS (pos.): m/z 464 (M+H) +ESI-MS (neg.): m/z 462 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 5×10 -3M (CD 50?5×10 -5M)
CEM-SS 5×10 -3M (CD 50?8×10 -6M)
MT4 7×10 -4M (CD 50?3×10 -6M)
Embodiment 13: preparation 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) dichloro thiophosphoryl phenylalanine methyl ester compound, wherein R is C 6H 5CH 2, B is a cytosine(Cyt).
The structural formula of compound is:
Figure A0113078400141
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the phenylalanine methyl ester hydrochloride of 1mmol (0.22g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene cytidine synthetic with above-mentioned embodiment 11.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.282g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene cytidine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl phenylalanine methyl ester, productive rate is 56.7%.
Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 58.71,58.13; 1H NMR (500MHz, D 2O): δ 8.22,8.21 (1H, d, 3J=5.5, H-6), 7.29-7.41 (5H, m, Ph), 7.03 (1H, m, H-1 '), 5.84,5.83 (1H, d, 3J=6, H-5), 4.66 (1H, m, H-3 '), 3.82 (1H, m, H-4 '), 3.58 (3H, s, OCH 3), 3.44 (2H, m, H-5 '), 2.89 (2H, m, H-α), 2.66 (1H, H-2 '), 2.27 (1H, m, H-β), 1.15 (3H, s, CH 3), 1.07 (3H, s, CH 3); 13C NMR (500MHz, D 2O): δ 180.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 146.36 (Ph-jpso), 117.02,136.11 (C-6), 137.12 (Ph-para), 136.42 (Ph-ortho), 122.91 (Ph-meta), 121.11 (＞CMe 2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH 3), 50.86 (C-β), 45.89 (C-α), 39.82 (C-2 '), 27.66 (CH 3), 24.15 (CH 3); ESI-MS (pos.): m/z 540 (M+H) +ESI-MS (neg.): m/z 538 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 5×10 -4M (CD 50?8×10 -5M)
CEM-SS 6×10 -4M (CD 50?8×10 -5M)
MT4 8×10 -4M (CD 50?7×10 -6M)
Embodiment 14: preparation 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester compound, wherein R is (CH 3) 2CH 2, B is a cytosine(Cyt).
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the valine methyl ester hydrochloride of 1mmol (0.17g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene cytidine synthetic with above-mentioned embodiment 11.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.282g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene cytidine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester, productive rate is 52.4%.
Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 56.98,56.21; 1H NMR (500MHz, D 2O): δ 7.90,7.89 (1H, d, 3J=5.5, H-6), 6.63 (1H, m, H-1 '), 5.84,5.83 (1H, d, 3J=6, H-5), 4.61 (1H, m, H-3 '), 3.69 (1H, m, H-4 '), 3.76 (3H, s, OCH 3), 3.65 (2H, m, H-5 '), 2.89 (2H, m, H-α), 2.66 (1H, H-2 '), 2.14 (1H, m, H-β), 1.15 (3H, s, CH 3), 1.07 (3H, s, CH 3); 13C NMR (500MHz, D 2O): δ 185.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 145.34 (C-6), 121.11 (＞CMe 2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH 3), 50.86 (C-β), 49.89 (C-α), 46.21 (C-β), 39.82 (C-2 '), 24.37 (CH 3), 23.22 (CH 3); ESI-MS (pos.): m/z 506 (M+H) +ESI-MS (neg.): m/z 504 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 5×10 -4M (CD 50?5×10 -7M)
CEM-SS 5×10 -4M (CD 50?6×10 -6M)
MT4 8×10 -3M (CD 50?3×10 -5M)
Embodiment 15.2 ', 3 '-O-isopropylidene cytidine 5 '-preparation of (isopropoxy replacement) thiophosphoryl leucine methyl compound, wherein R is (CH 3) 2CHCH 2, B is a cytosine(Cyt).
The structural formula of compound is:
Figure A0113078400161
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the leucine methyl ester hydrochloride of 1mmol (0.18g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene cytidine synthetic with above-mentioned embodiment 11.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.282g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene cytidine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl leucine methyl esters, productive rate is 55.4%.
Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 54.36,53.56; 1H NMR (500MHz, D 2O): δ 7.96,7.95 (1H, d, 3J=6, H-6), 6.36 (1H, m, H-1 '), 5.84,5.83 (1H, d, 3J=6, H-5), 4.78 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.76 (3H, s, OCH 3), 3.65 (2H, m, H-5 '), 3.22 (2H, m, H-α), 2.96 (1H, H-2 '), 2.77 (1H, m, H-β), 2.39 (1H, m, H-γ), 1.15 (3H, s, CH 3), 1.07 (3H, s, CH 3); 13C NMR (500MHz, D 2O): δ 185.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 145.34 (C-6), 121.11 (＞CMe 2), 93.71 (C-5), 88.25 (C-1 '), 85.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.88 (OCH 3), 50.86 (C-β), 48.89 (C-α), 46.21 (C-γ), 39.82 (C-2 '), 23.39 (CH 3), 23.01 (CH 3); ESI-MS (pos.): m/z 508 (M+H) +ESI-MS (neg.): m/z 506 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 5×10 -3M (CD 50?6×10 -6M)
CEM-SS 8×10 -4M (CD 50?9×10 -6M)
MT4 7×10 -4M (CD 50?8×10 -6M)
Embodiment 16: preparation 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl glycine methyl ester compound, wherein R is H, B is a uridylic.The structural formula of compound is: The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the glycine methyl ester hydrochloride of 1mmol (0.125g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) preparation 2 ', 3 '-O-isopropylidene uridine is standby
Get 1mol (244g) uridine under the room temperature and be suspended in the acetone of 2L, stir down to wherein adding 1.1mol (210g) tosic acid and 4mol (640ml) triethyl orthoformate, system change clarification after about 1 hour, adding 1mol (84g) NaHCO 3Neutralize, continue to stir 15 minutes, filter the back solids washed with acetone, with obtain behind filtrate and the washings evaporate to dryness product 2 ', 3 '-O-isopropylidene uridine slowly separates out, standing over night under the room temperature, after the filtration again with ice-water mixture thorough washing, filter rear filtrate through underpressure distillation obtain product 2 ', 3 '-O-isopropylidene uridine 256g (90%).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, 1mmol (0.284g) has been dissolved in 2 among the THF ', 3 '-O-isopropylidene uridine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl glycine methyl ester, productive rate is 63.4%.
Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 57.68,56.26; 1H NMR (500MHz, D 2O): δ 7.90,7.89 (1H, d, 3J=5.5, H-6), 6.24 (1H, m, H-1 '), 5.84,5.83 (1H, d, 3J=6, H-5), 4.28 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.71 (3H, s, OCH 3), 3.65 (2H, m, H-5 '), 2.69 (2H, m, H-α), 2.15 (1H, H-2 '), 1.15 (3H, s, CH 3), 1.07 (3H, s, CH 3); 13C NMR (500MHz, D 2O): δ 177.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 140.34 (C-6), 121.11 (＞CMe 2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH 3), 45.89 (C-α), 39.82 (C-2 '), 28.66 (CH 3), 26.15 (CH 3); ESI-MS (pos.): m/z 480 (M+H) +ESI-MS (neg.): m/z 478 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 8×10 -3M (CD 50?9×10 -5M)
CEM-SS 9×10 -3M (CD 50?8×10 -5M)
MT4 8×10 -4M (CD 50?5×10 -6M)
Embodiment 17: preparation 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl alanine methyl ester compound, wherein R is CH 3, B is a uridylic.
The structural formula of compound is:
Figure A0113078400172
The synthesis step of compound:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the alanine methyl ester hydrochloride of 1mmol (0.14g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene uridine synthetic with above-mentioned experiment 16.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, 1mmol (0.284g) has been dissolved in 2 among the THF ', 3 '-O-isopropylidene uridine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl alanine methyl ester, productive rate is 58.4%.Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 58.32,57.68; 1H NMR (500MHz, D 2O): δ 7.90,7.89 (1H, d, 3J=5.5, H-6), 6.24 (1H, m, H-1 '), 5.84,5.83 (1H, d, 3J=6, H-5), 4.28 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.71 (3H, s, OCH 3), 3.65 (2H, m, H-5 '), 2.69 (2H, m, H-α), 2.15 (1H, H-2 '), 1.15 (3H, s, CH 3), 1.07 (3H, s, CH 3); 13C NMR (500MHz, D 2O): δ 177.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 140.34 (C-6), 121.11 (＞CMe 2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH 3), 45.89 (C-α), 39.82 (C-2 '), 28.66 (CH 3), 26.15 (CH 3); ESI-MS (pos.): m/z 466 (M+H) +ESI-MS (neg.): m/z 464 (M-H) -. the anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 8×10 -4M (CD 50?9×10 -4M)
CEM-SS 7×10 -4M (CD 50?6×10 -5M)
MT4 8×10 -3M (CD 50?5×10 -5M)
Embodiment 18.2 ', 3 '-O-isopropylidene uridine 5 '-preparation of (isopropoxy replacement) thiophosphoryl alanine methyl ester compound, wherein R is C 6H 5CH 2, B is a uridylic.The structural formula of compound:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the phenylalanine methyl ester hydrochloride of 1mmol (0.21g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene uridine synthetic with above-mentioned experiment 16.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, 1mmol (0.284g) has been dissolved in 2 among the THF ', 3 '-O-isopropylidene uridine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl phenylalanine methyl ester, productive rate is 51.4%.Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 60.32,59.68; 1H NMR (500MHz, D 2O): δ 7.90,7.89 (1H, d, 3J=5.5, H-6), 6.24 (1H, m, H-1 '), 5.84,5.83 (1H, d, 3J=6, H-5), 4.28 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.71 (3H, s, OCH 3), 3.65 (2H, m, H-5 '), 2.69 (2H, m, H-α), 2.15 (1H, H-2 '), 1.15 (3H, s, CH 3), 1.07 (3H, s, CH 3); 13C NMR (500MHz, D 2O): δ 177.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 140.34 (C-6), 121.11 (＞CMe 2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH 3), 45.89 (C-α), 39.82 (C-2 '), 28.66 (CH 3), 26.15 (CH 3); ESI-MS (pos.): m/z 542 (M+H) +ESI-MS (neg.): m/z 540 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 6×10 -4M (CD 50?7×10 -5M)
CEM-SS 9×10 -3M (CD 50?9×10 -5M)
MT4 5×10 -3M (CD 50?6×10 -4M)
Embodiment 19: preparation 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester compound, wherein R is (CH 3) 2CH 2, B is a uridylic.
The structural formula of compound is:
Figure A0113078400191
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the valine methyl ester hydrochloride of 1mmol (0.17g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene uridine synthetic with above-mentioned experiment 16.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, 1mmol (0.284g) has been dissolved in 2 among the THF ', 3 '-O-isopropylidene uridine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester, productive rate is 66.4%.
Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 60.82,59.98; 1H NMR (500MHz, D 2O): δ 7.90,7.89 (1H, d, 3J=5.5, H-6), 6.24 (1H, m, H-1 '), 5.84,5.83 (1H, d, 3J=6, H-5), 4.28 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.71 (3H, s, OCH 3), 3.65 (2H, m, H-5 '), 2.69 (2H, m, H-α), 2.15 (1H, H-2 '), 1.15 (3H, s, CH 3), 1.07 (3H, s, CH 3); 13C NMR (500MHz, D 2O): δ 177.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 140.34 (C-6), 121.11 (＞CMe 2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH 3), 45.89 (C-α), 39.82 (C-2 '), 28.66 (CH 3), 26.15 (CH 3); ESI-MS (pos.): m/z 542 (M+H) +ESI-MS (neg.): m/z 540 (M-H) -. the anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 6×10 -4M (CD 50?7×10 -5M)
CEM-SS 9×10 -3M (CD 50?9×10 -5M)
MT4 5×10 -3M (CD 50?6×10 -4M)
Embodiment 20: preparation 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl leucine methyl compound, wherein R is (CH 3) 2CHCH 2, B is a uridylic.
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under the nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the leucine methyl ester hydrochloride of 1mmol (0.18g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O diisopropylidene uridine synthetic with above-mentioned experiment 16.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, lmmol (0.284g) has been dissolved in 2 among the THF ', 3 '-O-isopropylidene uridine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl leucine methyl esters, productive rate is 62.4%.
Spectral data is as follows: 31P NMR (D 2O, δ: ppm, J:Hz): δ 59.87,59.68; 1H NMR (500MHz, D 2O): δ 7.90,7.89 (1H, d, 3J=5.5, H-6), 6.24 (1H, m, H-1 '), 5.84,5.83 (1H, d, 3J=6, H-5), 4.28 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.71 (3H, s, OCH 3), 3.65 (2H, m, H-5 '), 2.69 (2H, m, H-α), 2.15 (1H, H-2 '), 1.15 (3H, s, CH 3), 1.07 (3H, s, CH 3); 13C NMR (500MHz, D 2O): δ 177.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 140.34 (C-6), 121.11 (＞CMe 2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH 3), 45.89 (C-α), 39.82 (C-2 '), 28.66 (CH 3), 26.15 (CH 3); ESI-MS (pos.): m/z 542 (M+H) +ESI-MS (neg.): m/z 540 (M-H) -.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED 50?CEM-TK- 6×10 -4M (CD 50?7×10 -5M)
CEM-SS 9×10 -3M (CD 50?9×10 -5M)
MT4 5×10 -3M (CD 50?6×10 -4M)

Claims (2)

1, a kind of 2 ', 3 '-O-isopropylidene nucleosides 5 '-(isopropoxy replacement) thiophosphoryl amino acid ester compound, it is characterized in that the structural formula of this compound is: R is H, CH in the said structure formula 3, C 6H 5CH 2, (CH 3) 2CH 2Or (CH 3) 2CHCH 2In any; B is any in VITAMIN B4, guanine, cytosine(Cyt) or the uridylic.
2, a kind of preparation method of compound as claimed in claim 1 is characterized in that this method comprises the steps:
1) under nitrogen protection, cryosel is bathed under the condition that is cooled to-4～-8 ℃, and raw material (isopropoxy replacement) two compd 22/190s are dissolved in the dry tetrahydrofuran (THF) of crossing, and is mixed with the solution that concentration is 0.8～1.0mol/L;
2) add amino acid methyl ester hydrochloride with the amount of raw material same substance in above-mentioned solution, slowly drip acid binding agent second triamine after stirring, the add-on of acid binding agent is 2 times of amount of raw material;
3) preparation 2 ', 3 '-O-isopropylidene nucleosides solution for standby:
Get nucleosides under the room temperature and be suspended in the solution that is made into 0.5～1.0mol/L in the exsiccant solvent, stir down to wherein adding and the tosic acid of amount of substances such as nucleosides and the triethyl orthoformate of four times of amount of substances, after system becomes clarification, neutralize with big water gaging with strong aqua and to make PH=7～8, then above-mentioned solution decompression is concentrated, in refrigerator standing over night can obtain product 2 ', 3 '-O-isopropylidene nucleosides.With make 2 ', 3 '-be dissolved in the solution that is mixed with 0.5～1.0 mol/L in the solvent after the O-isopropylidene nucleosides suction filtration drying;
4) follow the tracks of above-mentioned second reaction process that goes on foot, after treating that the raw material total overall reaction finishes, will with 2 of above-mentioned the 3rd step preparation of the amount of raw material same substance ', 3 '-O-isopropylidene nucleosides solution slowly splashes in the solution in second step, and the back that stirs continue to drip the acid binding agent with the amount of raw material same substance;
5) after monitoring reaction is finished, filter, rotary distillation removes and desolvates and other low-boiling point materials, be hydrolyzed with ammoniacal liquor at last, recycle silicon glue post carries out column chromatography for separation, can obtain 2 ', 3 '-O-isopropylidene nucleosides 5 '-(isopropoxy replacement) thiophosphoryl amino acid ester compound.
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Cited By (4)

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US8871737B2 (en) 2010-09-22 2014-10-28 Alios Biopharma, Inc. Substituted nucleotide analogs
US8916538B2 (en) 2012-03-21 2014-12-23 Vertex Pharmaceuticals Incorporated Solid forms of a thiophosphoramidate nucleotide prodrug
US8980865B2 (en) 2011-12-22 2015-03-17 Alios Biopharma, Inc. Substituted nucleotide analogs
US9012427B2 (en) 2012-03-22 2015-04-21 Alios Biopharma, Inc. Pharmaceutical combinations comprising a thionucleotide analog

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8871737B2 (en) 2010-09-22 2014-10-28 Alios Biopharma, Inc. Substituted nucleotide analogs
US9278990B2 (en) 2010-09-22 2016-03-08 Alios Biopharma, Inc. Substituted nucleotide analogs
US8980865B2 (en) 2011-12-22 2015-03-17 Alios Biopharma, Inc. Substituted nucleotide analogs
US9605018B2 (en) 2011-12-22 2017-03-28 Alios Biopharma, Inc. Substituted nucleotide analogs
US8916538B2 (en) 2012-03-21 2014-12-23 Vertex Pharmaceuticals Incorporated Solid forms of a thiophosphoramidate nucleotide prodrug
US9394330B2 (en) 2012-03-21 2016-07-19 Alios Biopharma, Inc. Solid forms of a thiophosphoramidate nucleotide prodrug
US9856284B2 (en) 2012-03-21 2018-01-02 Alios Biopharma, Inc. Solid forms of a thiophosphoramidate nucleotide prodrug
US9012427B2 (en) 2012-03-22 2015-04-21 Alios Biopharma, Inc. Pharmaceutical combinations comprising a thionucleotide analog

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