CN1290707A - Nucleoside 5'-thiophosphoryl amino-acid ester compound - Google Patents
Nucleoside 5'-thiophosphoryl amino-acid ester compound Download PDFInfo
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- CN1290707A CN1290707A CN 00129555 CN00129555A CN1290707A CN 1290707 A CN1290707 A CN 1290707A CN 00129555 CN00129555 CN 00129555 CN 00129555 A CN00129555 A CN 00129555A CN 1290707 A CN1290707 A CN 1290707A
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- 239000002777 nucleoside Substances 0.000 title abstract description 6
- 150000003833 nucleoside derivatives Chemical class 0.000 title abstract description 6
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims abstract description 158
- 238000006243 chemical reaction Methods 0.000 claims abstract description 85
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims abstract description 78
- 150000001875 compounds Chemical class 0.000 claims abstract description 75
- 238000003756 stirring Methods 0.000 claims abstract description 57
- 235000011114 ammonium hydroxide Nutrition 0.000 claims abstract description 49
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 29
- 238000009835 boiling Methods 0.000 claims abstract description 29
- 238000004440 column chromatography Methods 0.000 claims abstract description 29
- 238000004821 distillation Methods 0.000 claims abstract description 29
- 239000000741 silica gel Substances 0.000 claims abstract description 29
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 29
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000002253 acid Substances 0.000 claims abstract description 17
- 239000011230 binding agent Substances 0.000 claims abstract description 17
- 239000000126 substance Substances 0.000 claims abstract description 13
- 150000001413 amino acids Chemical class 0.000 claims abstract description 9
- 150000004702 methyl esters Chemical class 0.000 claims abstract description 9
- 239000002904 solvent Substances 0.000 claims abstract 5
- 238000005481 NMR spectroscopy Methods 0.000 claims description 76
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 56
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 52
- NYHBQMYGNKIUIF-UUOKFMHZSA-N Guanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NYHBQMYGNKIUIF-UUOKFMHZSA-N 0.000 claims description 38
- DRTQHJPVMGBUCF-XVFCMESISA-N Uridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-XVFCMESISA-N 0.000 claims description 38
- 230000007062 hydrolysis Effects 0.000 claims description 32
- 238000006460 hydrolysis reaction Methods 0.000 claims description 32
- 229910052757 nitrogen Inorganic materials 0.000 claims description 28
- 238000000926 separation method Methods 0.000 claims description 28
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 26
- UHDGCWIWMRVCDJ-UHFFFAOYSA-N 1-beta-D-Xylofuranosyl-NH-Cytosine Natural products O=C1N=C(N)C=CN1C1C(O)C(O)C(CO)O1 UHDGCWIWMRVCDJ-UHFFFAOYSA-N 0.000 claims description 19
- MIKUYHXYGGJMLM-GIMIYPNGSA-N Crotonoside Natural products C1=NC2=C(N)NC(=O)N=C2N1[C@H]1O[C@@H](CO)[C@H](O)[C@@H]1O MIKUYHXYGGJMLM-GIMIYPNGSA-N 0.000 claims description 19
- UHDGCWIWMRVCDJ-PSQAKQOGSA-N Cytidine Natural products O=C1N=C(N)C=CN1[C@@H]1[C@@H](O)[C@@H](O)[C@H](CO)O1 UHDGCWIWMRVCDJ-PSQAKQOGSA-N 0.000 claims description 19
- NYHBQMYGNKIUIF-UHFFFAOYSA-N D-guanosine Natural products C1=2NC(N)=NC(=O)C=2N=CN1C1OC(CO)C(O)C1O NYHBQMYGNKIUIF-UHFFFAOYSA-N 0.000 claims description 19
- DRTQHJPVMGBUCF-PSQAKQOGSA-N beta-L-uridine Natural products O[C@H]1[C@@H](O)[C@H](CO)O[C@@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-PSQAKQOGSA-N 0.000 claims description 19
- UHDGCWIWMRVCDJ-ZAKLUEHWSA-N cytidine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O1 UHDGCWIWMRVCDJ-ZAKLUEHWSA-N 0.000 claims description 19
- 229940029575 guanosine Drugs 0.000 claims description 19
- DRTQHJPVMGBUCF-UHFFFAOYSA-N uracil arabinoside Natural products OC1C(O)C(CO)OC1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-UHFFFAOYSA-N 0.000 claims description 19
- 229940045145 uridine Drugs 0.000 claims description 19
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 claims description 14
- MWEQTWJABOLLOS-UHFFFAOYSA-L disodium;[[[5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-oxidophosphoryl] hydrogen phosphate;trihydrate Chemical compound O.O.O.[Na+].[Na+].C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP([O-])(=O)OP(O)([O-])=O)C(O)C1O MWEQTWJABOLLOS-UHFFFAOYSA-L 0.000 claims description 12
- 239000002126 C01EB10 - Adenosine Substances 0.000 claims description 7
- 229960005305 adenosine Drugs 0.000 claims description 7
- FAIAAWCVCHQXDN-UHFFFAOYSA-N phosphorus trichloride Chemical compound ClP(Cl)Cl FAIAAWCVCHQXDN-UHFFFAOYSA-N 0.000 claims 8
- GVNVAWHJIKLAGL-UHFFFAOYSA-N 2-(cyclohexen-1-yl)cyclohexan-1-one Chemical compound O=C1CCCCC1C1=CCCCC1 GVNVAWHJIKLAGL-UHFFFAOYSA-N 0.000 claims 4
- 101150065749 Churc1 gene Proteins 0.000 claims 4
- 102100038239 Protein Churchill Human genes 0.000 claims 4
- 238000002360 preparation method Methods 0.000 abstract description 21
- 230000000840 anti-viral effect Effects 0.000 abstract description 6
- 230000000259 anti-tumor effect Effects 0.000 abstract description 5
- 230000036436 anti-hiv Effects 0.000 abstract description 3
- 239000003814 drug Substances 0.000 abstract description 3
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 abstract 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 abstract 1
- 229940079593 drug Drugs 0.000 abstract 1
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 129
- WQYSXVGEZYESBR-UHFFFAOYSA-N thiophosphoryl chloride Chemical compound ClP(Cl)(Cl)=S WQYSXVGEZYESBR-UHFFFAOYSA-N 0.000 description 48
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 40
- 230000000694 effects Effects 0.000 description 26
- 230000015572 biosynthetic process Effects 0.000 description 24
- 239000000463 material Substances 0.000 description 24
- 238000012544 monitoring process Methods 0.000 description 24
- 229960001866 silicon dioxide Drugs 0.000 description 24
- 238000003786 synthesis reaction Methods 0.000 description 24
- 238000002474 experimental method Methods 0.000 description 22
- 238000005160 1H NMR spectroscopy Methods 0.000 description 20
- 238000004679 31P NMR spectroscopy Methods 0.000 description 20
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical group CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 20
- 239000003480 eluent Substances 0.000 description 20
- 125000001160 methoxycarbonyl group Chemical group [H]C([H])([H])OC(*)=O 0.000 description 20
- 230000003595 spectral effect Effects 0.000 description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 20
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 19
- 235000001014 amino acid Nutrition 0.000 description 13
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 8
- 241000725303 Human immunodeficiency virus Species 0.000 description 5
- DWKPPFQULDPWHX-VKHMYHEASA-N l-alanyl ester Chemical compound COC(=O)[C@H](C)N DWKPPFQULDPWHX-VKHMYHEASA-N 0.000 description 5
- MCTWTZJPVLRJOU-UHFFFAOYSA-N 1-methyl-1H-imidazole Chemical compound CN1C=CN=C1 MCTWTZJPVLRJOU-UHFFFAOYSA-N 0.000 description 4
- COQRGFWWJBEXRC-UHFFFAOYSA-N hydron;methyl 2-aminoacetate;chloride Chemical compound Cl.COC(=O)CN COQRGFWWJBEXRC-UHFFFAOYSA-N 0.000 description 4
- SWVMLNPDTIFDDY-FVGYRXGTSA-N methyl (2s)-2-amino-3-phenylpropanoate;hydrochloride Chemical compound Cl.COC(=O)[C@@H](N)CC1=CC=CC=C1 SWVMLNPDTIFDDY-FVGYRXGTSA-N 0.000 description 4
- QVDXUKJJGUSGLS-LURJTMIESA-N methyl L-leucinate Chemical compound COC(=O)[C@@H](N)CC(C)C QVDXUKJJGUSGLS-LURJTMIESA-N 0.000 description 4
- RYYWUUFWQRZTIU-UHFFFAOYSA-N Thiophosphoric acid Chemical compound OP(O)(S)=O RYYWUUFWQRZTIU-UHFFFAOYSA-N 0.000 description 3
- KUGLDBMQKZTXPW-JEDNCBNOSA-N methyl (2s)-2-amino-3-methylbutanoate;hydrochloride Chemical compound Cl.COC(=O)[C@@H](N)C(C)C KUGLDBMQKZTXPW-JEDNCBNOSA-N 0.000 description 3
- 101710163270 Nuclease Proteins 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 150000004713 phosphodiesters Chemical class 0.000 description 2
- 239000000651 prodrug Substances 0.000 description 2
- 229940002612 prodrug Drugs 0.000 description 2
- 108060002716 Exonuclease Proteins 0.000 description 1
- 206010061598 Immunodeficiency Diseases 0.000 description 1
- 208000029462 Immunodeficiency disease Diseases 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 239000000074 antisense oligonucleotide Substances 0.000 description 1
- 238000012230 antisense oligonucleotides Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 102000013165 exonuclease Human genes 0.000 description 1
- 230000007813 immunodeficiency Effects 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 125000004437 phosphorous atom Chemical group 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003582 thiophosphoric acids Chemical class 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Saccharide Compounds (AREA)
Abstract
本发明涉及一种核苷5’－硫代磷酰氨基酸酯化合物,其结构式为右式,制备方法为:首先将三氯硫磷溶于已干燥过的四氢呋喃中,加入氨基酸甲酯盐酸盐,搅拌后滴加缚酸剂,反应完成后,过滤,旋转蒸馏除去溶剂及其他低沸点物质,最后用氨水进行水解,用硅胶柱进行柱层析分离,即可得到本发明的产物。本发明的化合物,可用于开发抗病毒、抗肿瘤、抗HIV活性药物。
The invention relates to a nucleoside 5'-thiophosphoryl amino acid ester compound, the structural formula of which is the right formula, and the preparation method is as follows: firstly, thion trichloride is dissolved in dried tetrahydrofuran, and amino acid methyl ester hydrochloride is added , after stirring, add acid-binding agent dropwise, after the reaction is completed, filter, remove solvent and other low boiling point substances by rotary distillation, finally hydrolyze with ammonia water, and separate by column chromatography with silica gel column to obtain the product of the present invention. The compound of the present invention can be used for developing antiviral, antitumor and antiHIV active drugs.
Description
The present invention relates to the novel nucleoside 5 '-thiophosphoryl amino-acid ester compound of a class, this compounds has pharmaceutical activitys such as good biological activity and antiviral, antitumor, anti-HIV, can develop into antiviral, antitumor, the anti-HIV prodrug of a kind of ucleosides (prodrugs) in clinical application.
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.
The objective of the invention is to develop a class nucleosides-thiophosphoryl amino acid ester compound, by synthetic thiophosphoric acid compounds with opposing nuclease, synthetic dna oligo is to be used to develop antiviral, antitumor, HIV (human immunodeficiency virus)-resistant activity medicine.
Total following four kinds of the nucleoside 5 '-thiophosphoryl amino-acid ester compound of the present invention's preparation,
Wherein first kind is adenosine 5 '-thiophosphoryl amino acid ester compound, and 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
2
The synthesis step of above-claimed cpd: 1) under nitrogen protection, room temperature (25 ℃) is dissolved in phosphorus thiochloride in the dry tetrahydrofuran (THF) of crossing (THF), is mixed with the solution of 1mol/L.2) the amino acid methyl ester hydrochloride of adding same substance amount in above-mentioned solution slowly drips acid binding agent (as triethylamine, the N-Methylimidazole) after stirring.3) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the adenosine that is dissolved in the dry pyridine is slowly splashed in the above-mentioned system, acid binding agent 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.5) after the hydrolysis fully, carry out column chromatography for separation, can obtain product with silicagel column.
Second kind of compound wherein is guanosine 5 '-thiophosphoryl amino acid ester compound, and 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
3
The synthesis step of above-claimed cpd:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in phosphorus thiochloride in the dry tetrahydrofuran (THF) of crossing (THF), is mixed with the solution of 1mol/L.
2) the amino acid methyl ester hydrochloride of the amount of adding same substance in above-mentioned solution slowly drips acid binding agent (as triethylamine, the N-Methylimidazole) after stirring.
3) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the guanosine that is dissolved in the dry pyridine is slowly splashed in the above-mentioned system, acid binding agent 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.
5) after the hydrolysis fully, carry out column chromatography for separation, can obtain product with silicagel column.
The third compound wherein is cytidine 5 '-thiophosphoryl amino acid ester compound, and 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
2
The synthesis step of above-claimed cpd:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in phosphorus thiochloride in the dry tetrahydrofuran (THF) of crossing (THF), is mixed with the solution of 1mol/L.
2) the amino acid methyl ester hydrochloride of the amount of adding same substance in above-mentioned solution slowly drips acid binding agent (as triethylamine, the N-Methylimidazole) after stirring.
3) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the cytidine that is dissolved in the dry pyridine is slowly splashed in the above-mentioned system, acid binding agent 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.
5) after the hydrolysis fully, carry out column chromatography for separation, can obtain product with silicagel column.
The 4th kind of compound wherein is uridine 5 '-thiophosphoryl amino acid ester compound, and 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
2
The synthesis step of above-claimed cpd:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in phosphorus thiochloride in the dry tetrahydrofuran (THF) of crossing (THF), is mixed with the solution of 1mol/L.
2) the amino acid methyl ester hydrochloride of the amount of adding same substance in above-mentioned solution slowly drips acid binding agent (as three second peace, N-Methylimidazole) after stirring.
3) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the uridine that is dissolved in the dry THF is slowly splashed in the above-mentioned system, acid binding agent 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.
5) after the hydrolysis fully, carry out column chromatography for separation, can obtain product with silicagel column.
The anti-HIV-1 activity experiment of above-mentioned synthetic series compound in cem cell and MT-4 cell finds all to have activity in various degree, can be used to develop antiviral, antitumor, HIV (human immunodeficiency virus)-resistant activity medicine.
Below be embodiments of the invention:
Embodiment 1: the preparation of adenosine 5 '-thiophosphoryl glycine methyl ester compound, wherein R is H.
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), 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) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the adenosine that 1mmol (0.267g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product adenosine 5 '-thiophosphoryl glycine methyl ester at 16.5: 1: 1, productive rate is 68.4%.Spectral data is as follows:
31P NMR (DMSO-d
6, δ: ppm, J:Hz): δ 59.38,58.62;
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)；
13C?NMR(500MHz，DMSO-d
6)：δ173．24(COOMe)，163．70(C-2)，150．43(C-4)，140．76(C-6)，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)；ESI-MS(pos．)：m／z?435(M+H)
+；ESI-MS(neg．)：m/z?433(M-H)
-。The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
HIV=Human?immunodeficiency?virus
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
-4M (CD
50?7×10
-6M)
CEM-SS 4×10
-3M (CD
50?9×10
-5M)
MT?4 2×10
-1M (CD
50 8×10
-6M)
Embodiment 2: the preparation of adenosine 5 '-thiophosphoryl alanine methyl ester compound, wherein R is CH3.The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) 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) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the adenosine that 1mmol (0.267g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product adenosine 5 '-thiophosphoryl alanine methyl ester at 16.5: 1: 1, productive rate is 66.4%.Spectral data is as follows:
31P NMR (DMSO-d
6, δ: ppm, J:Hz): δ 59.86,58.53;
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), 1.31,1.30 (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), l49.76 (C-6), (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); ESI-MS (pos.): m/z 449 (M+H)
+ESI-MS (neg.): m/z 447 (M-H)
-The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 8×10
-3M (CD
50 3×10
-6M)
CEM-SS 6×10
-3M (CD
50 9×10
-5M)
MT?4 8×10
-4M (CD
50 6×10
-6M)
Embodiment 3: the preparation of adenosine 5 '-thiophosphoryl phenylalanine methyl ester compound, wherein R is C
6H
5CH
2The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), 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) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the adenosine that 1mmol (0.267g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product adenosine 5 '-thiophosphoryl phenylalanine methyl ester at 16.5: 1: 1, productive rate is 69.7%.Spectral data is as follows:
31P NMR (DMSO-d
6, δ: ppm, J:Hz): δ 60.64,60.33;
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);
13C NMR (500MHz, DMSO-d
6): δ 174.24 (COOMe), 170.60 (C-2), 159.43 (C-4), 148.36 (Ph-Jpso), 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); ESI-MS (pos.): m/z 527 (M+H)
+ESI-MS (neg.): m/z 525 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 8×10
-2M (CD
50 4×10
-5M)
CEM-SS 9×10
-3M (CD
50 4×10
-6M)
MT?4 6×10
-5M (CD
50 5×10
-6M)
Embodiment 4: the preparation of adenosine 5 '-thiophosphoryl valine methyl ester compound, wherein R is (CH
3)
2CHCH.
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), 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 reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the adenosine that 1mmol (0.267g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product adenosine 5 '-thiophosphoryl valine methyl ester at 16.5: 1: 1, productive rate is 59.2%.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);
13C NMR (500MHz, DMSO-d
6): δ 172.24 (COOMe), 165.70 (C-2), 159.43 (C-4), 145.76 (C-6), (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-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 7×10
-3M (CD
50 8×l0
-6M)
CEM-SS 6×10
-3M (CD
50 5×10
-6M)
MT?4 8×10
-4M (CD
50 6×10
-5M)
Embodiment 5: the preparation of adenosine 5 '-thiophosphoryl leucine methyl compound, wherein R is (CH
3)
2CHCH
2
The structural formula of compound:
The synthesis step of compound
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), 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 reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the adenosine that 1mmol (0.267g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product adenosine 5 '-thiophosphoryl leucine methyl esters at 16.5: 1: 1, productive rate is 65.2%.Spectral data is as follows:
31P NMR (DMSO-d
6, δ: ppm, J:Hz): δ 58.32,57.68;
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);
13C NMR (500MHz, DMSO-d
6): δ 172.24 (COOMe), 165.70 (C-2), 159.43 (C-4), 145.76 (C-6), (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); ESI-MS (pos.): m/z 492 (M+H)
+ESI-MS (neg.): m/z 490 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 5×10
-4M (CD
50 2×10
-6M)
CEM-SS 2×10
-4M (CD
50 5×10
-5M)
MT?4 5×10
-3M (CD
50 8×10
-6M)
Embodiment 6: the preparation of guanosine 5 '-thiophosphoryl glycine methyl ester compound, wherein R is H.
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), 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) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the guanosine that 1mmol (0.283g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product guanosine 5 '-thiophosphoryl glycine methyl ester at 16.5: 1: 1, productive rate is 63.4%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 57.68,57.39;
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);
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), 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); ESI-MS (pos.): m/z 451 (M+H)
+ESI-MS (neg.): m/z 449 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 8×10
-4M (CD
50 6×10
-5M)
CEM-SS 3×10
-4M (CD
50 4×10
-6M)
MT?4 6×10
-4M (CD
50 3×10
-5M)
Embodiment 7: the preparation of guanosine 5 '-thiophosphoryl alanine methyl ester compound, wherein R is CH
3
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) 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 (0.140g), slowly drip 2mmol (0.2g) triethylamine after stirring.
4) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the guanosine that 1mmol (0.283g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
6) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product guanosine 5 '-thiophosphoryl alanine methyl ester at 16.5: 1: 1, productive rate is 65.3%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 58.88,58.37;
1H NMR (500MHz, D
2O): δ 8.07 (1H, s, H-8), 5.82,5.81 (1H, d,
1J=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);
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), 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); ESI-MS (pos.): m/z 466 (M+H)
+ESI-MS (neg.): m/z 464 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 6×10
-4M (CD
50 7×10
-5M)
CEM-SS 9×10
-3M (CD
50 6×10
-5M)
MT?4 2×10
-4M (CD
50 8×10
-5M)
Embodiment 8: the preparation of guanosine 5 '-thiophosphoryl phenylalanine methyl ester compound, wherein R is C
6H
5CH
2
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), 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) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the guanosine that 1mmol (0.283g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product guanosine 5 '-thiophosphoryl phenylalanine methyl ester at 16.5: 1: 1, productive rate is 70.3%.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);
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), 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); ESI-MS (pos.): m/z 541 (M+H)
+ESI-MS (neg.): m/z 539 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 8×10
-3M (CD
50 6×10
-5M)
CEM-SS 6×10
-4M (CD
50 5×10
-5M)
MT?4 2×10
-4M (CD
50 8×10
-5M)
Embodiment 9: the preparation of guanosine 5 '-thiophosphoryl valine methyl ester compound, wherein R is (CH
3)
2CH.
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) 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 (0.170g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the guanosine that 1mmol (0.283g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product guanosine 5 '-thiophosphoryl valine methyl ester at 16.5: 1: 1, productive rate is 66.2%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 59.86,59.27;
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), 1.16,1.15 (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), 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); ESI-MS (pos.): m/z 493 (M+H)
+ESI-MS (neg.): m/z 491 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 5×10
-3M (CD
50 3×10
-5M)
CEM-SS 5×10
-3M (CD
50 8×10
-5M)
MT?4 4×10
-3M (CD
50 6×10
-5M)
Embodiment 10: the preparation of guanosine 5 '-thiophosphoryl leucine methyl compound, wherein R is (CH
3)
2CHCH
2
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), 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) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the guanosine that 1mmol (0.283g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product guanosine 5 '-thiophosphoryl leucine methyl esters at 16.5: 1: 1, productive rate is 62.2%.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), 3.22 (1H, m, H-γ), 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), 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); ESI-MS (pos.): m/z 508 (M+H)
+ESI-MS (neg.): m/z 506 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 6×10
-4M (CD
50 6×10
-5M)
CEM-SS 6×10
-4M (CD
50 5×10
-5M)
MT?4 8×10
-4M (CD
50 7×10
-5M)
Embodiment 11: the preparation of cytidine 5 '-thiophosphoryl glycine methyl ester compound, wherein R is H.
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), 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) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the cytidine that 1mmol (0.242g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product cytidine 5 '-thiophosphoryl glycine methyl ester at 16.5: 1: 1, productive rate is 61.4%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 57.88,56.16;
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 ');
13C NMR (500MHz, D
2O): δ 177.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 140.34 (C-6), 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 '); ESI-MS (pos.): m/z 411 (M+H)
+ESI-MS (neg.): m/z 409 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 8×10
-4M (CD
50 6×10
-5M)
CEM-SS 6×10
-3M (CD
50 4×10
-5M)
MT?4 6×10
-4M (CD
50 3×10
-6M)
Embodiment 12: the preparation of cytidine 5 '-thiophosphoryl alanine methyl ester compound, wherein R is CH
3
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) 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.
4) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the cytidine that 1mmol (0.242g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
6) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product cytidine 5 '-thiophosphoryl alanine methyl ester at 16.5: 1: 1, productive rate is 68.4%.Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 57.66,57.31;
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);
13C NMR (500MHz, D
2O): δ 185.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 145.34 (C-6), 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 '); ESI-MS (pos.): m/z 424 (M+H)
+ESI-MS (neg.): m/z 422 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 5×10
-3M (CD
50 5×10
-5M)
CEM-SS 5×10
-3M (CD
50 8×10
-6M)
MT?4 7×10
-4M (CD
50 3×10
-6M)
Embodiment 13: the system of cytidine 5 '-thiophosphoryl phenylalanine methyl ester compound is put forth energy, and wherein R is C
6H
5CH
2
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), 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) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the cytidine that 1mmol (0.242g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product cytidine 5 '-thiophosphoryl phenylalanine methyl ester at 16.5: 1: 1, productive rate is 66.7%.
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);
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), 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 '); ESI-MS (pos.): m/z 500 (M+H)
+ESI-MS (neg.): m/z 498 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 5×10
-4M (CD
50 8×10
-5M)
CEM-SS 6×10
-4M (CD
50 8×10
-5M)
MT?4 8×10
-4M (CD
50 7×10
-5M)
Embodiment 14: the preparation of cytidine 5 '-thiophosphoryl valine methyl ester compound, wherein R is (CH
3)
2CH.
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), 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 reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the cytidine that 1mmol (0.242g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product cytidine 5 '-thiophosphoryl valine methyl ester at 16.5: 1: 1, productive rate is 63.4%.Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 59.98,59.21;
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);
13C NMR (500MHz, D
2O): δ 185.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 145.34 (C-6), 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), ESI-MS (pos.): m/z 466 (M+H)
+ESI-MS (neg.): m/z 464 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 5×10
-4M (CD
50 5×10
-5M)
CEM-SS 5×10
-4M (CD
50 9×10
-5M)
MT?4 8×10
-3M (CD
50 3×10
-5M)
Embodiment 15: the preparation of cytidine 5 '-thiophosphoryl leucine methyl compound, wherein R is (CH
3)
2CHCH
2
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), 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 reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the cytidine that 1mmol (0.242g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product cytidine 5 '-thiophosphoryl leucine methyl esters at 16.5: 1: 1, productive rate is 65.4%.Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 56.36,56.56;
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);
13C NMR (500MHz, D
2O): δ 185.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 145.34 (C-6), 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 '); ESI-MS (pos.): m/z 468 (M+H)
+ESI-MS (neg.): m/z 466 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 5×10
-3M (CD
50 6×10
-6M)
CEM-SS 8×10
-3M (CD
50 9×10
-6M)
MT?4 7×10
-3M (CD
50 8×10
-6M)
Embodiment 16: the preparation of uridine 5 '-thiophosphoryl glycine methyl ester compound, wherein R is H.
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), 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) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the uridine that 1mmol (0.244g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product uridine 5 '-thiophosphoryl glycine methyl ester at 16.5: 1: 1, productive rate is 68.4%.Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 57.68,56.89;
1H NMR (500MHz, D
2O): δ 7.79,7.78 (1H, dd,
3J=4.5, H-6), 5.82 (2H, m, H-1 ', 5), 4.93 (2H, m, H-2 ', 3 '), 4.52 (1H, m, H-4 '), 4.00 (2H, m, H-5 '), 3.63 (3H, s, OCH
3), 3.57 (2H, m, H-α);
13C NMR (500MHz, D
2O): δ 177.08 (COOMe), 169.02 (C-4), 153.99 (C-2), 145.39,145.36 (C-6), 104.33,104.22 (C-5), 95.83,95.16 (C-1 '), (87.97 C-4 '), 87.20,87.13 (C-2 '), 83.64, (83.51 C-3 '), 67.24 (C-5 '), 55.09 (OCH
3), 45.80,45.73 (C-α); ESI-MS (pos.): m/z 413 (M+H)
+ESI-MS (neg.): m/z 411 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 8×10
-3M (CD
50 9×10
-5M)
CEM-SS 9×10
-3M (CD
50 8×10
-5M)
MT?4 8×10
-4M (CD
50 5×10
-6M)
Embodiment 17: the preparation of uridine 5 '-thiophosphoryl alanine methyl ester compound, wherein R is CH
3
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) 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) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the uridine that 1mmol (0.244g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product uridine 5 '-thiophosphoryl alanine methyl ester at 16.5: 1: 1, productive rate is 71.4%.Spectral data is as follows: 31P NMR (D
2O, δ: ppm, J:Hz): δ 58.32,57.68;
1H NMR (500MHz, D
2O): δ 7.88,7.87 (1H, dd,
3J=5, H-6), 5.89 (2H, m, H-1 ', 5), 4.98 (2H, m, H-2 ', 3 '), 4.61 (1H, m, H-4 '), 4.04 (2H, m, H-5 '), 3.73 (3H, s, OCH
3), 3.57 (2H, m, H-α), 1.31,1.30 (3H, d,
3J=6, β-CH
3);
13C NMR (500MHz, D
2O): δ 175.04 (COOMe), 166.73 (C-4), 149.28 (C-2), 144.36,144.32 (C-6), 107.62,107.43 (C-5), 98.76,98.41 (C-1 '), (86.23 C-4 '), 85.71,85.63 (C-2 '), 82.70, (82.58 C-3 '), 65.44 (C-5 '), 56.87 (OCH
3), 50.73 (C-α), 48.66 (C-β); ESI-MS (pos.): m/z 426 (M+H)
+ESI-MS (neg.): m/z 424 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 8×10
-4M (CD
50 6×10
-5M)
CEM-SS 7×10
-4M (CD
50 6×10
-5M)
MT?4 8×10
-4M (CD
50 5×10
-5M)
Embodiment 18: the preparation of uridine 5 '-thiophosphoryl phenylalanine methyl ester compound, wherein R is C
6H
5CH
2
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), 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) follow the tracks of reaction process with nuclear magnetic resonance analyser (1NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the uridine that 1mmol (0.244g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product uridine 5 '-thiophosphoryl phenylalanine methyl ester at 16.5: 1: 1, productive rate is 72.4%.Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 60.32,59.68;
1H NMR (500MHz, D
2O): δ 7.88,7.87 (1H, dd,
3J=5, H-6), 7.22-7.46 (5H, m, Ph), 6.08 (2H, m, H-1 ', 5), 5.15 (2H, m, H-2 ', 3 '), 4.71 (1H, m, H-4 '), 4.34 (2H, m, H-5 '), 3.93 (3H, s, OCH
3), 3.57 (2H, m, H-α), 2.37 (2H, m, H-β);
13C NMR (500MHz, D
2O): δ 178.04 (COOMe), 166.73 (C-4), 149.28 (C-2), 148.39 (Ph-jpso), 148.36,148.32 (C-6), 139.12 (Ph-para), 129.42 (Ph-ortho), 122.91 (Ph-meta), 110.62,110.43 (C-5), 98.76,98.41 (C-1 '), 88.23 (C-4 '), 85.71,85.63 (C-2 '), 83.27, (82.58 C-3 '), 65.44 (C-5 '), 56.87 (OCH
3), 50.73 (C-α), 48.66 (C-β); ESI-MS (pos.): m/z 522 (M+H)
+ESI-MS (neg.): m/z 520 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 8×10
-4M (CD
50 7×10
-5M)
CEM-SS 7×10
-4M (CD
50 6×10
-5M)
MT?4 6×10
-4M (CD
50 6×10
-5M)
Embodiment 19: the preparation of uridine 5 '-thiophosphoryl valine methyl ester compound, wherein R is (CH
3)
2CH.
The structural formula of compound:
The synthesis step of compound
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the valine methyl ester hydrochloride of 1mmol (0.14g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the uridine that 1mmol (0.244g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product uridine 5 '-thiophosphoryl valine methyl ester at 16.5: 1: 1, productive rate is 65.4%.Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 58.32,57.55;
1H NMR (500MHz, D
2O): δ 7.89,7.88 (1H, dd,
3J=5, H-6), 5.99 (2H, m, H-1 ', 5), 4.96 (2H, m, H-2 ', 3 '), 4.61 (1H, m, H-4 '), 4.04 (2H, m, H-5 '), 3.73 (3H, s, OCH
3), 3.57 (2H, m, H-α), 3.45 (1H, m, H-β), 1.31,1.30 (2H, d, 2CH
3);
13C NMR (500MHz, D
2O): δ 178.04 (COOMe), 167.55 (C-4), 146.28 (C-2), 144.36,144.32 (C-6), 107.62,107.43 (C-5), 98.76,98.41 (C-1 '), (86.23 C-4 '), 85.71,85.63 (C-2 '), 82.70, (82.58 C-3 '), 65.44 (C-5 '), 56.87 (OCH
3), 50.73 (C-α), 48.66 (C-β), 22.45 (CH
3), 22.55 (CH
3); ESI-MS (pos.): m/z 454 (M+H)
+ESI-MS (neg.): m/z 452 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 9×10
-3M (CD
50 8×10
-5M)
CEM-SS 6×10
-3M (CD
50 8×10
-5M)
MT?4 8×10
-4M (CD
50 2×10
-5M)
Embodiment 20: the preparation of uridine 5 '-thiophosphoryl leucine methyl compound, wherein R is (CH
3)
2CHCH
2
The structural formula of compound:
The synthesis step of compound:
1) under nitrogen protection, room temperature (25 ℃) is dissolved in the phosphorus thiochloride of 1mmol (0.17g) in the dry tetrahydrofuran (THF) of crossing (THF), 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 reaction process with nuclear magnetic resonance analyser (NMR), treat that the phosphorus thiochloride total overall reaction finishes after, the uridine that 1mmol (0.244g) has been dissolved in the dry pyridine slowly splashes in the above-mentioned system, 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.
5) after the hydrolysis fully, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=can obtain product uridine 5 '-thiophosphoryl leucine methyl esters at 16.5: 1: 1, productive rate is 67.5%.Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 59.66,59.10;
1H NMR (500MHz, D2O): δ 7.89,7.88 (1H, dd,
3J=5, H-6), 5.82 (2H, m, H-1 ', 5), 4.86 (2H, m, H-2 ', 3 '), 4.41 (1H, m, H-4 '), 4.04 (2H, m, H-5 '), 3.73 (3H, s, OCH
3), 3.57 (2H, m, H-α), 3.45 (1H, m, H-β), 3.21 (2H, m, H-γ), 1.31,1.30 (2H, d, 2CH
3);
13C NMR (500MHz, D
2O): δ 178.04 (COOMe), 167.55 (C-4), 152.28 (C-2), 146.36,146.32 (C-6), 107.62,107.43 (C-5), 98.76,98.41 (C-1 '), (88.23 C-4 '), 85.71,85.63 (C-2 '), 82.70, (82.58 C-3 '), 67.44 (C-5 '), 58.54 (OCH
3), 50.73 (C-α), 48.66 (C-β), 35.68 (C-γ), 22.45 (CH
3), 22.55 (CH
3); ESI-MS (pos.): m/z 468 (M+H)
+ESI-MS (neg.): m/z 466 (M-H)
-
The anti-HIV-1 activity experiment of this compound in cem cell and MT-4 cell
ED
50 CEM-TK- 5×10
-5M (CD
50 6×10
-4M)
CEM-SS 6×10
-5M (CD
50 4×10
-5M)
MT?4 7×10
-5M (CD
50 7×10
-5M)
Nucleoside 5 '-thiophosphoryl amino acid methyl ester be a class brand-new have an active compound of inverase, by synthetic different types of nucleosides-amino acid conjugate, and carry out activity experiment, tentatively obtain satisfied result, be further development and the development of new inverase achievement in research that provides the foundation.
Claims (4)
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| CNB001295551A CN1133642C (en) | 2000-10-09 | 2000-10-09 | Nucleoside 5'-phosphorothioate amino acid ester compound |
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|---|---|---|---|
| CNB001295551A CN1133642C (en) | 2000-10-09 | 2000-10-09 | Nucleoside 5'-phosphorothioate amino acid ester compound |
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| WO2012053917A1 (en) | 2010-10-19 | 2012-04-26 | Instytut Chemii Bioorganicznej Pan | Nucleotide analogue, method of synthesis of nucleotide analogue, use of nucleotide analogue, antiviral pro-nucleotide, pharmaceutical composition |
| CN103209987A (en) * | 2010-09-22 | 2013-07-17 | 艾丽奥斯生物制药有限公司 | Substituted nucleotide analogs |
| US8916538B2 (en) | 2012-03-21 | 2014-12-23 | Vertex Pharmaceuticals Incorporated | Solid forms of a thiophosphoramidate nucleotide prodrug |
| CN104334570A (en) * | 2012-03-21 | 2015-02-04 | 艾丽奥斯生物制药有限公司 | Methods of preparing substituted nucleotide analogs |
| 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 |
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