CN1343673A - Thiophosphamino acid ester compound containing 3'-azido-deoxythymidine and its preparing process - Google Patents

Abstract

The invention relates to a thiophosphoryl amino acid ester compound containing 3' azidothymidine and a preparation method thereof. The structure of the compound is the right formula. Its preparation method is to dissolve the raw material (aryl group substituted) dichlorphosphion in tetrahydrofuran (THF), add amino acid methyl ester hydrochloride, then add acid-binding agent dropwise, and then add 3' azidothymidine solution to the above solution Adding an acid-binding agent, filtering, rotary distillation, and separation by column chromatography with a silica gel column to obtain 3'azidothymidine-5'-(aryl substituted) thiophosphoryl amino acid ester compound. The compound prepared by the method of the present invention has anti-AIDS virus drug activity.

Description

A thiophosphoryl amino acid ester compound containing 3' azidothymidine and its preparation method

The invention relates to a thiophosphoryl amino acid ester compound and a preparation method thereof, in particular to a thiophosphoryl amino acid ester compound containing 3' azidothymidine and a preparation method thereof. This type of compound has good biological activity and antiviral properties , anti-tumor, anti-AIDS virus and other drug activities, can be developed into a nucleoside anti-virus, anti-tumor, anti-AIDS virus prodrugs (prodrugs) in clinical application.

Thiophosphoric acid contains a structure in which sulfur replaces oxygen on the phosphorus atom. Although this structure still retains the original charge and high water solubility, other oligodeoxyribonucleic acid (S-oligos) of phosphorothioate Both the physicochemical and biological properties are quite different from the natural phosphodiester prototype. One of the most notable differences is that phosphorothioates are resistant to nucleases. Due to the presence of intracellular and extracellular nucleases, the added phosphodiester antisense oligonucleotides are quickly digested, making them lose their function. By synthesizing phosphorothioate compounds with resistance to nucleases, there is no doubt that a promising path has been opened for the future application of synthetic oligodeoxyribonucleic acid for therapeutic purposes.

The object of the present invention is to propose a thiophosphoryl amino acid ester compound containing 3' azidethymidine and a preparation method thereof, which are used for developing antiviral, antitumor and anti-AIDS virus active drugs.

上述结构式中R＝H，CH₃，C₆H₅CH₂，(CH₃)₂CH₂，(CH₃)₂CHCH₂；The thiophosphoryl amino acid ester compound containing 3' azidothymidine prepared by the present invention has a structural formula of:

In the above structural formula, R=H, CH ₃ , C ₆ H ₅ CH ₂ , (CH ₃ ) ₂ CH ₂ , (CH ₃ ) ₂ CHCH ₂ ;

X = H, Cl, _NO2 .

The synthetic steps of above-mentioned compound are as follows:

1) Under the protection of nitrogen, under the condition of cooling to -4~-8°C in an ice-salt bath, dissolve the raw material (aryl group substituted) parathion dichloride in dried tetrahydrofuran (THF) to prepare a concentration of 0.8 ~1.0mol/L solution.

2) Add amino acid methyl ester hydrochloride in the same amount as the raw material to the above solution, stir evenly and then slowly add the acid-binding agent dropwise, the amount of the acid-binding agent added is twice the amount of the raw material.

3) Track the above reaction process. After all the raw materials have been reacted, slowly drop the 3' azidethymidine solution dissolved in tetrahydrofuran into the solution of the second step, stir evenly and continue Add dropwise the acid-binding agent of the same amount as the raw material.

4) After monitoring the completion of the reaction, filter, remove the solvent and other low boiling point substances by rotary distillation, and carry out column chromatography separation with a silica gel column to obtain a thiophosphoryl amino acid ester compound containing 3' azidothymidine.

The thiophosphoryl amino acid ester compound containing 3' azidethymidine prepared by the method of the present invention is a new type of compound with anti-AIDS drug activity, through the synthesis of different types of nucleoside-amino acid conjugates , and carried out activity experiments, and initially obtained satisfactory results, which provided basic research results for the further research and development of new anti-AIDS virus drugs. The anti-AIDS virus-1 activity experiments in CEM cells and MT-4 cells found that they all have different degrees of activity.

Introduce the embodiment of the present invention below:

Example 1: Preparation of 3'azidovidine-5'-(phenyl substituted)thiophosphorylglycine methyl ester compound, wherein R=H, X=H.

The structural formula of the compound is:

The synthetic steps of compound are as follows:

1) Under the protection of nitrogen, cool in an ice-salt bath to -4 ~ -8C, dissolve 1mmol (0.227g) of (phenyl substituted) dichloride dichloride in dried tetrahydrofuran (THF), and prepare 1mol/ L solution.

2) Add 1 mmol (0.125 g) of glycine methyl ester hydrochloride to the above solution, stir well and slowly add 2 mmol (0.2 g) triethylamine dropwise.

3) Track the reaction process with nuclear magnetic resonance (NMR). After the (phenyl substituted) dichloride dichloride is completely reacted, slowly drop 1mmol (0.267g) of 3' azidothymidine dissolved in THF into In the above system, continue to add 1 mmol (0.1 g) of triethylamine dropwise after stirring evenly.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(phenyl substituted)phosphorothioglycine methyl ester, the yield was 77.1%.

Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ71.70, 72.10; ¹ H NMR (500 MHz, DMSO-d ₆ ): δ9.80 (1H, sb, NH, 7.22-7.41 (6H, m, Ph, H-6), 6.32 (1H, m, H-1′), 4.20-4.41 (4H, H-3′, H-4′, H-5′), 4.01 (1H, m, Ala-NH), 3.88 (3H, s, OCH ₃ ), 3.57 (2H, m, H-α), 2, 50 (1H, m, H-2′), 2.02 (3H, s , 5-Me); ¹³ C NMR (500MHz, DMSO-d ₆ ): δ 173.24 (COOMe), 163.70 (C-2), 150.43 (C-4), 149.33 (d, Ph-ipso, J=4Hz) , 140.76(C-6), 129.91(m, Ph-para), 125.52(m, Ph-ortho), 120.08(m, Ph-meta), 111.52(C-5), 84.94(C-1′), 82.40(C-4'), 65.87(C-5'), 60.54(C-3'), 54.78(OCH ₃ ), 50.41(C-α), 37.46(C-2'), 12.60(5-Me ); ESI-MS (pos.): m/z 511 (M+H) ⁺ ; ESI-MS (neg.): m/z 509 (MH) ^- . The compound in CEM cells and MT-4 cells Anti-AIDS virus-1 activity test

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 9×10 ^-6 μM (CD ₅₀ 206μM)

CEM-SS 4×10 ^-5 μM (CD ₅₀ 118μM)

MT4 5×10 ^-5 μM (CD ₅₀ 70μM)

Example 2: Preparation of 3'azidovidine-5'-(phenyl substituted)thiophosphorylalanine methyl ester compound, wherein R is CH ₃ and X is H.

The structural formula of the compound:

The synthetic steps of compound are as follows:

1) Under the protection of nitrogen, cool in an ice-salt bath to -4 ~ -8 ° C, dissolve 1 mmol (0.227 g) of (phenyl substituted) dichloride dichloride in dried tetrahydrofuran (THF), and prepare 1 mol /L solution.

2) Add 1 mmol (0.14 g) of alanine methyl ester hydrochloride to the above solution, stir well and slowly add 2 mmol (0.2 g) of triethylamine dropwise.

3) Track the reaction process with nuclear magnetic resonance (NMR). After the (phenyl substituted) dichloride dichloride is completely reacted, slowly drop 1mmol (0.267g) of 3' azidothymidine dissolved in THF into In the above system, continue to add 1 mmol (0.1 g) of triethylamine dropwise after stirring evenly.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(phenyl substituted)thiophosphorylalanine methyl ester, the yield was 67.3%.

Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ70.20, 71.70; ¹ H NMR (500 MHz, DMSO-d ₆ ): δ9.81 (1H, sb, NH) , 7.22-7.41 (6H, m, Ph, H-6), 6.34 (1H, m, H-1'), 4.20-4.41 (4H, H-3', H-4', H-5'), 4.01 (1H, m, Ala-NH), 3.88 (3H, s, OCH ₃ ), 3.56 (2H, m, H-α), 2, 50 (1H, m, H-2′), 2.02 (3H, s, 5-Me), 1.40 (3H, d, Ala-Me); ¹³ C NMR (500MHz, DMSO-d ₆ ): δ 178.24 (COOMe), 163.70 (C-2), 150.43 (C-4), 149.33 (d, Ph-ipso, J=4Hz), 140.76 (C-6), 129.91 (m, Ph-para), 125.52 (m, Ph-ortho), 120.08 (m, Ph-meta), 111.52 (C -5), 84.94(C-1′), 82.40(C-4′), 65.87(C-5′), 60.54(C-3′), 54.78(OCH ₃ ), 50.41(C-α), 37.46 (C-2'), 20.86 (d, Ala-Me, J=5Hz), 12.60 (5-Me); ESI-MS (pos.): m/z 525 (M+H) ⁺ ; ESI-MS (neg .): m/z523(MH) ^- Anti-AIDS virus-1 activity test of the compound in CEM cells and MT-4 cells

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 6××10 ^-3 μM (CD ₅₀ 188μM)

CEM-SS 6××10 ^-2 μM (CD ₅₀ 49μM)

MT4 4××10 ^-2 μM (CD ₅₀ 69μM)

Example 3: Preparation of 3'azidovidine-5'-(phenyl substituted)thiophosphoryl phenylalanine methyl ester compound, wherein R is C ₆ H ₅ CH ₂ , X is H.

The structural formula of the compound:

The synthetic steps of compound are as follows:

1) Under the protection of nitrogen, cool in an ice-salt bath to -4 ~ -8C, dissolve 1mmol (0.227g) of (phenyl substituted) dichloride dichloride in dried tetrahydrofuran (THF), and prepare 1mol/ L solution.

2) Add 1 mmol (0.215 g) of phenylalanine methyl ester hydrochloride to the above solution, stir well and slowly add 2 mmol (0.2 g) triethylamine dropwise.

3) Use nuclear magnetic resonance (NMR) to track the reaction process. After the (phenyl substituted) thionyl dichloride has completely reacted, slowly drop 1mmol (0.266g) of 3' azidothymidine dissolved in THF into In the above system, continue to add 1 mmol (0.1 g) of triethylamine dropwise after stirring evenly.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(phenyl substituted) phosphorothioate phenylalanine methyl ester, the yield was 69.6%.

Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ70.70, 71.30; ¹ H NMR (500 MHz, DMSO-d ₆ ): δ9.88 (1H, sb, NH) , 7.22-7.41 (6H, m, Ph, H-6), 7.15-7.35 (5H, m, Ph), 6.32 (1H, m, H-1′), 4.20-4.44 (4H, H-3′, H-4', H-5'), 4.01 (1H, m, Ala-NH), 3.88 (3H, s, OCH ₃ ), 3.57 (2H, m, H-α), 2,50 (1H, m , H-2'), 2.32 (2H, m, H-β), 2.02 (3H, s, 5-Me); ¹³ C NMR (500MHz, DMSO-d ₆ ): δ 174.39 (COOMe), 163.66 (C -2), 151.24(C-4), 148.36(Ph-pso), 140.76(C-6), 120.08(m, Ph-meta), 135.12(Ph-para), 130.42(Ph-ortho), 111.52( C-5), 84.94(C-1'), 82.40(C-4'), 65.87(C-5'), 60.54(C-3'), 56.82(C-β), 54.78(OCH ₃ ),

50.41 (C-α), 37.46 (C-2′), 12.60 (5-Me);

ESI-MS (pos.): m/z 601 (M+H) ⁺ ; ESI-MS (neg.): m/z 599 (MH) ^-

Anti-AIDS virus-1 activity experiment of the compound in CEM cells and MT-4 cells

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 8×10 ^-3 μM (CD ₅₀ 112μM)

CEM-SS 3×10 ^-2 μM (CD ₅₀ 29μM)

MT4 6×10 ^-2 μM (CD ₅₀ 44μM)

Example 4: Preparation of 3'azidethymidine-5'-(phenyl substituted)thiophosphorylvaline methyl ester compound, wherein R is (CH ₃ ) ₂ CH ₂ , and X is H.

The structural formula of the compound:

Compound synthesis steps:

1) Under the protection of nitrogen, cool in an ice-salt bath to -4 ~ -8 ° C, dissolve 1 mmol (0.227 g) of (phenyl substituted) dichloride dichloride in dried tetrahydrofuran (THF), and prepare 1 mol /L solution.

2) Add 1mmol (0.17g) of valine methyl ester hydrochloride to the above solution, stir well and slowly add 2mmol (0.2g) triethylamine dropwise.

3) Use nuclear magnetic resonance (NMR) to track the reaction process. After the (phenyl substituted) dichloride dichloride is completely reacted, slowly drop 1mmol (0.31g) of 3' azidothymidine dissolved in THF into In the above system, continue to add 1 mmol (0.1 g) of triethylamine dropwise after stirring evenly.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(phenyl substituted) phosphorothiovaline methyl ester, the yield was 87.3%.

Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ71.70, 72.40; ¹ H NMR (500 MHz, DMSO-d ₆ ): δ9.80 (1H, sb, NH) , 7.22-7.41 (6H, m, Ph, H-6), 6.32 (1H, m, H-1'), 4.20-4.41 (4H, H-3', H-4', H-5'), 4.01 (1H, m, Ala-NH), 3.88 (3H, s, OCH ₃ ), 3.67 (1H, m, H-α), 3.45 (1H, m, H-β), 2, 50 (1H, m , H-2'), 2.02 (3H, s, 5-Me), 1.61 (3H, s, CH ₃ ), 1.42 (3H, s, CH ₃ ); ¹³ C NMR (500MHz, DMSO-d ₆ ): δ174.24 (COOMe), 165.70 (C-2), 150.63 (C-4), 149.33 (d, Ph-ipso, J=4Hz), 140.76 (C-6), 129.91 (m, Ph-para), 125.52 (m, Ph-ortho), 120.08 (m, Ph-meta, 111.52 (C-5), 84.94 (C-1′), 82.40 (C-4′), 65.87 (C-5′), 60.54 ( C-3′), 54.78(OCH ₃ ), 50.41(C-α), 48.66(C-β), 26.86(CH ₃ ), 26.40(CH ₃ ), 37.46(C-2′), 12.60(5- Me); ESI-MS (pos.): m/z 553 (M+H) ⁺ ; ESI-MS (neg.): m/z 551 (MH) ^- : the compound is in CEM cells and MT-4 cells Anti-AIDS virus-1 activity test

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 6×10 ^-3 μM (CD ₅₀ 77μM)

CEM-SS 1×10 ^-2 μM (CD ₅₀ 48μM)

MT4 2×10 ^-2 μM (CD ₅₀ 62μM)

Example 5: Preparation of 3'azidethymidine-5'-(phenyl substituted)thiophosphorylleucine methyl ester compound, wherein R is (CH ₃ ) ₂ CHCH ₂ , and X is H.

The structural formula of the compound:

The synthetic steps of compound are as follows:

1) Under the protection of nitrogen, cool in an ice-salt bath to -4 ~ -8 ° C, dissolve 1 mmol (0.227 g) of (phenyl substituted) dichloride dichloride in dried tetrahydrofuran (THF), and prepare 1 mol /L solution.

2) Add 1 mmol (0.184 g) of leucine methyl ester hydrochloride to the above solution, stir well and slowly add 2 mmol (0.2 g) of triethylamine dropwise.

3) Use nuclear magnetic resonance (NMR) to track the reaction process. After the (phenyl substituted) dichloride dichloride is completely reacted, slowly drop 1mmol (0.31g) of 3' azidothymidine dissolved in THF into In the above system, continue to add 1 mmol (0.1 g) of triethylamine dropwise after stirring evenly.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(phenyl substituted)phosphorylleucine methyl ester with a yield of 68.3%. Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ70.6, 71.8; ¹ H NMR (500 MHz, DMSO-d ₆ ): δ9.80 (1H, sb, NH) , 7.22-7.41 (6H, m, Ph, H-6), 6.32 (1H, m, H-1'), 4.20-4.41 (4H, H-3', H-4', H-5'), 4.01 (1H, m, Ala-NH), 3.88 (3H, s, OCH ₃ ), 3.57 (2H, m, H-α), 3.45 (1H, m, H-β), 3.21 (1H, m, H -γ), 2, 50 (1H, m, H-2'), 2.02 (3H, s, 5-Me), 1.61 (3H, s, CH ₃ ), 1.42 (3H, s, CH ₃ ); ¹³ C NMR (500MHz, DMSO-d ₆ ): δ173.24 (COOMe), 163.70 (C-2), 150.43 (C-4), 149.33 (d, Ph-ipso, J=4Hz), 140.76 (C-6 ), 129.91(m, Ph-para), 125.52(m, Ph-ortho), 120.08(m, Ph-meta), 111.52(C-5), 84.94(C-1′), 82.40(C-4′ ), 65.87(C-5′), 60.54(C-3′), 54.78(OCH ₃ ), 50.41(C-α), 48.66(C-β), 46.35(C-γ), 23.40(CH ₃ ) , 22.86 (CH ₃ ), 37.46 (C-2′), 12.60 (5-Me); ESI-MS (pos.): m/z 567 (M+H) ⁺ ; ESI-MS (neg.): m /z 565(MH) ^- .

Anti-AIDS virus-1 activity experiment of the compound in CEM cells and MT-4 cells

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 8×10 ^-3 μM (CD ₅₀ 79μM)

CEM-SS 9×10 ^-2 μM (CD ₅₀ 78μM

MT4 5×10 ^-2 μM (CD ₅₀ 23μM)

Example 6: Preparation of 3'azidovidine-5'-(p-chlorophenyl substituted)phosphorothioglycine methyl ester compound, wherein R is H and X is CI.

The structural formula of the compound:

The synthetic steps of compound are as follows:

1) Under the protection of nitrogen, cool in an ice-salt bath to -4~-8C, dissolve 1mmol (0.262g) of (p-chlorophenyl substituted) dichlorphosphos in dried tetrahydrofuran (THF), and prepare 1mol/L solution.

2) Add 1 mmol (0.125 g) of glycine methyl ester hydrochloride to the above solution, stir well and slowly add 2 mmol (0.2 g) triethylamine dropwise.

3) track the reaction process with a nuclear magnetic resonance instrument (NMR), and after the (p-chlorophenyl substituted) parathionium dichloride has completely reacted, slowly dissolve 1mmol (0.267g) of 3' azidothymidine dissolved in THF Drop into the above system, stir evenly and continue to drop 1mmol (0.1g) triethylamine.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(p-chlorophenyl substituted)phosphorothioglycine methyl ester, the yield was 77.1%.

Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ71.32, 72.11; ¹ H NMR (500 MHz, DMSO-d ₆ : δ9.80 (1H, sb, NH), 6.82-7.25 (5H, m, Ph, H-6), 6.32 (1H, m, H-1′), 4.20-4.41 (4H, H-3′, H-4′, H-5′), 4.01 (1H, m, Ala-NH), 3.88 (3H, s, OCH ₃ ), 3.57 (2H, m, H-α), 2, 50 (1H, m, H-2′), 2.02 (3H, s , 5-Me); ¹³ C NMR (500MHz, DMSO-d ₆ ): δ 173.24 (COOMe), 163.70 (C-2), 159.91 (m, Ph-para), 150.43 (C-4), 149.33 (d , Ph-ipso, J=4Hz), 140.76 (C-6), 125.52 (m, Ph-ortho), 117.08 (m, Ph-meta), 111.52 (C-5), 84.94 (C-1′), 82.40(C-4'), 65.87(C-5'), 60.54(C-3'), 54.78(OCH ₃ ), 50.41(C-α), 37.46(C-2'), 12.60(5-Me ); ESI-MS (pos.): m/z547 (M+H) ⁺ ; ESI-MS (neg.): m/z545 (MH) ^- . The anti-love of the compound in CEM cells and MT-4 cells ZV-1 activity test

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 8×10 ^-3 μM (CD ₅₀ 186μM)

CEM-SS 5×10 ^-2 μM (CD ₅₀ 218μM)

MT4 5×10 ^-2 μM (CD ₅₀ 60μM)

Example 7: Preparation of 3'azidovidine-5'-(p-chlorophenyl substituted)thiophosphorylalanine methyl ester compound, wherein R is CH ₃ , and X is CI.

The structural formula of the compound:

The synthetic steps of compound are as follows:

1) Under the protection of nitrogen, cool in an ice-salt bath to -4～-8°C, dissolve 1mmol (0.262g) of (p-chlorophenyl substituted)phosphorous dichloride in dried tetrahydrofuran (THF), and prepare into a 1mol/L solution.

2) Add 1 mmol (0.14 g) of alanine methyl ester hydrochloride to the above solution, stir well and slowly add 2 mmol (0.2 g) of triethylamine dropwise.

3) track the reaction process with a nuclear magnetic resonance instrument (NMR), and after the (p-chlorophenyl substituted) parathionium dichloride has completely reacted, slowly dissolve 1mmol (0.267g) of 3' azidothymidine dissolved in THF Drop into the above system, stir evenly and continue to drop 1mmol (0.1g) triethylamine.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(p-chlorophenyl substituted)thiophosphorylalanine methyl ester, the yield was 71.3%. Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ70.20, 71.70; ¹ H NMR (500 MHz, DMSO-d ₆ ): δ9.81 (1H, sb, NH) , 6.82-7.28 (5H, m, Ph, H-6), 6.36 (1H, m, H-1'), 4.20-4.41 (4H, H-3', H-4', H-5'), 4.11 (1H, m, Ala-NH), 3.86 (3H, s, OCH ₃ ), 3.66 (2H, m, H-α), 2,50 (1H, m, H-2′), 2.02 (3H, s, 5-Me), 1.40 (3H, d, Ala-Me); ¹³ C NMR (500MHz, DMSO-d ₆ : δ 177.24 (COOMe), 161.70 (C-2), 159.91 (m, Ph-para) , 150.43(C-4), 149.33(d, Ph-ipso, J=4Hz), 140.76(C-6), 125.52(m, Ph-ortho), 116.08(m, Ph-meta), 111.52(C- 5), 84.94(C-1′), 82.40(C-4′), 65.87(C-5′), 60.54(C-3′), 54.78(OCH ₃ ), 50.41(C-α), 35.46( C-2'), 19.86 (d, Ala-Me, J=5Hz), 12.60 (5-Me); ESI-MS (pos.): m/z 560 (M+H) ⁺ ; ESI-MS (neg .): m/z 558 MH) ^- .

Anti-AIDS virus-1 activity experiment of the compound in CEM cells and MT-4 cells

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 9×10 ^-3 μM (CD ₅₀ 188μM))

CEM-SS 5×10 ^-2 μM (CD ₅₀ 123μM)

MT4 6×10 ^-2 μM (CD ₅₀ 98μM)

Example 8: Preparation of 3'azidovidine-5'-(p-chlorophenyl substituted)thiophosphoryl phenylalanine methyl ester compound, wherein R is C ₆ H ₅ CH ₂ , and X is CI.

The structural formula of the compound:

The synthetic steps of compound are as follows:

1) Under nitrogen protection, cool in an ice-salt bath to -4~-8°C, dissolve 1 mmol (0.262 g) of (p-chlorophenyl substituted) dichlorphosphos in dried tetrahydrofuran (THF), and prepare into a 1mol/L solution.

2) Add 1 mmol (0.215 g) of phenylalanine methyl ester hydrochloride to the above solution, stir well and slowly add 2 mmol (0.2 g) triethylamine dropwise.

3) track the reaction process with a nuclear magnetic resonance instrument (NMR), and after the (p-chlorophenyl substituted) parathionium dichloride has completely reacted, slowly dissolve 1mmol (0.266g) of 3' azidothymidine dissolved in THF Drop into the above system, stir evenly and continue to drop 1mmol (0.1g) triethylamine.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(p-chlorophenyl substituted)thiophosphoryl phenylalanine methyl ester, the yield was 70.8%. Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ71.70, 72.30; ¹ H NMR (500 MHz, DMSO-d ₆ ): δ9.81 (1H, sb, NH) , 6.82-7.25 (5H, m, Ph, H-6), 7.15-7.35 (5H, m, Ph), 6.32 (1H, m, H-1′), 4.20-4.44 (4H, H-3′, H-4', H-5'), 4.01 (1H, m, Ala-NH), 3.88 (3H, s, OCH ₃ ), 3.57 (2H, m, H-α), 2, 50 (1H, m , H-2'), 2.32 (2H, m, H-β), 2.02 (3H, s, 5-Me); ¹³ C NMR (500MHz, DMSO-d ₆ : δ174.39 (COOMe), 163.66 (C -2), 159.91(m, Ph-para), 150.43(C-4), 149.33(d, Ph-ipso, J=4Hz), 140.76(C-6), 125.52(m, Ph-ortho), 117.08 (m, Ph-meta), 111.52 (C-5), 84.94 (C-1′), 82.40 (C-4′), 65.87 (C-5′), 60.54 (C-3′), 56.82 (C -β), 54.78 (OCH ₃ ), 50.41 (C-α), 37.46 (C-2′), 12.60 (5-Me); ESI-MS (pos.): m/z 636 (M+H) ⁺ ; ESI-MS (neg.): m/z 634 (MH) ^- .

Anti-AIDS virus-1 activity experiment of the compound in CEM cells and MT-4 cells

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 8×10 ^-3 μM (CD ₅₀ 185μM)

CEM-SS 5×10 ^-2 μM (CD ₅₀ 65μM)

MT4 6×10 ^-2 μM (CD ₅₀ 122μM)

Example 9: Preparation of 3'azidethymidine-5'-(p-chlorophenyl substituted)thiophosphorylvaline methyl ester compound, wherein R is (CH ₃ ) ₂ CH ₂ , and X is CI.

The structural formula of the compound:

The synthetic steps of compound are as follows:

1) Under nitrogen protection, cool in an ice-salt bath to -4~-8°C, dissolve 1 mmol (0.262 g) of (p-chlorophenyl substituted) dichlorphosphos in dried tetrahydrofuran (THF), and prepare into a 1mol/L solution.

2) Add 1mmol (0.17g) of valine methyl ester hydrochloride to the above solution, stir well and slowly add 2mmol (0.2g) triethylamine dropwise.

3) Track the reaction process with a nuclear magnetic resonance instrument (NMR). After the (p-chlorophenyl substituted) thionyl chloride has completely reacted, slowly dissolve 1mmol (0.31g) of 3' azidothymidine dissolved in THF Drop into the above system, stir evenly and continue to drop 1mmol (0.1g) triethylamine.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(p-chlorophenyl substituted)thiophosphoryl valine methyl ester, the yield was 67.3%.

Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ71.60, 72.50; ¹ H NMR (500 MHz, DMSO-d ₆ ): δ9.56 (1H, sb, NH) , 6.82-7.35 (5H, m, Ph, H-6), 6.32 (1H, m, H-1'), 4.20-4.41 (4H, H-3', H-4', H-5'), 4.01 (1H, m, Ala-NH), 3.88 (3H, s, OCH ₃ ), 3.69 (1H, m, H-α), 3.55 (1H, m, H-β), 2, 56 (1H, m , H-2'), 2.12 (3H, s, 5-Me), 1.51 (3H, s, CH ₃ ), 1.42 (3H, s, CH ₃ ); ¹³ C NMR (500MHz, DMSO-d ₆ ): δ175.24 (COOMe), 165.70 (C-2), 161.91 (m, Ph-para), 150.43 (C-4), 149.33 (d, Ph-ipso, J=4Hz), 140.76 (C-6), 125.52 (m, Ph-ortho), 117.08 (m, Ph-meta), 111.52 (C-5), 84.94 (C-1′), 82.40 (C-4′), 65.87 (C-5′), 60.54 (C-3'), 54.78(OCH ₃ ), 50.41(C-α), 48.66(C-β), 26.86(CH ₃ ), 26.40(CH ₃ ), 37.46(C-2'), 12.60(5 -Me); ESI-MS (pos.): m/z 587 (M+H) ⁺ ; ESI-MS (neg.): m/z 585 MH) ^- . Anti-AIDS virus-1 activity experiment of the compound in CEM cells and MT-4 cells

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 6×10 ^-3 μM (CD ₅₀ 77μM)

CEM-SS 1×10 ^-2 μM (CD ₅₀ 48μM)

MT4 2×10 ^-2 μM (CD ₅₀ 62μM)

Example 10: Preparation of 3'azidovidine-5'-(p-chlorophenyl substituted)thiophosphorylleucine methyl ester compound, wherein R is (CH ₃ ) ₂ CHCH ₂ , and X is CI.

The structural formula of the compound:

The synthetic steps of compound are as follows:

1) Under the protection of nitrogen, cool in an ice-salt bath to -4~-8C, dissolve 1mmol (0.262g) of (p-chlorophenyl substituted) dichlorphosphos in dried tetrahydrofuran (THF), and prepare 1mol/L solution.

2) Add 1 mmol (0.184 g) of leucine methyl ester hydrochloride to the above solution, stir well and slowly add 2 mmol (0.2 g) of triethylamine dropwise.

3) Track the reaction process with a nuclear magnetic resonance instrument (NMR). After the (p-chlorophenyl substituted) thionyl chloride has completely reacted, slowly dissolve 1mmol (0.31g) of 3' azidothymidine dissolved in THF Drop into the above system, stir evenly and continue to drop 1mmol (0.1g) triethylamine.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(p-chlorophenyl substituted)thiophosphorylleucine methyl ester, the yield was 66.3%. Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ71.8, 72.3; ¹ H NMR (500 MHz, DMSO-d ₆ ): δ9.85 (1H, sb, NH) , 6.82-7.21 (5H, m, Ph, H-6), 6.32 (1H, m, H-1′), 4.20-4.44 (4H, H-3′, H-4′, H-5′), 4.01 (1H, m, Ala-NH), 3.88 (3H, s, OCH ₃ ), 3.57 (2H, m, H-α), 3.45 (1H, m, H-β), 3.21 (1H, m, H -γ), 2, 50 (1H, m, H-2'), 2.02 (3H, s, 5-Me), 1.61 (3H, s, CH ₃ ), 1.42 (3H, s, CH ₃ ); ¹³ C NMR (500MHz, DMSO-d ₆ : δ 173.24 (COOMe), 163.70 (C-2), 159.91 (m, Ph-para), 150.43 (C-4), 149.33 (d, Ph-ipso, J= 4Hz), 140.76(C-6), 125.52(m, Ph-ortho), 117.08(m, Ph-meta), 111.52(C-5), 84.94(C-1′), 82.40(C-4′) , 65.87(C-5'), 60.54(C-3'), 54.78(OCH ₃ ), 50.41(C-α), 48.66(C-β), 46.35(C-γ), 23.40(CH ₃ ), 22.86 (CH ₃ ), 37.46 (C-2′), 12.60 (5-Me); ESI-MS (pos.): m/z 602 (M+H) ⁺ ; ESI-MS (neg.): m/ z 600(MH) ^- .The anti-AIDS virus-1 activity experiment of this compound in CEM cells and MT-4 cells

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 7×10 ^-3 μM (CD ₅₀ 159μM)

CEM-SS 9×10 ^-2 μM (CD ₅₀ 68μM)

MT4 6× ^10-2 μM (CD ₅₀ 21 μM)

Example 11: Preparation of 3'azidovidine-5'-(p-nitrophenyl substituted)thiophosphorylglycine methyl ester compound, wherein R is H and X is NO ₂ .

The structural formula of the compound:

The synthetic steps of compound are as follows:

1) Under the protection of nitrogen, cool in an ice-salt bath to -4～-8°C, dissolve 1 mmol (0.273 g) of (p-nitrobenzene substituted) dichloride phosphorus in dried tetrahydrofuran (THF), and prepare into a 1mol/L solution.

2) Add 1 mmol (0.125 g) of glycine methyl ester hydrochloride to the above solution, stir well and slowly add 2 mmol (0.2 g) triethylamine dropwise.

3) track the reaction process with a nuclear magnetic resonance instrument (NMR), and after the (p-nitrobenzene substituted) dichloride dichloride has completely reacted, slowly dissolve 1mmol (0.267g) of 3' azidothymidine dissolved in THF Drop into the above system, stir evenly and continue to drop 1mmol (0.1g) triethylamine.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(p-nitrophenyl substituted)phosphorothioglycine methyl ester, the yield was 66.1%.

Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ71.32, 72.11; ¹ H NMR (500 MHz, DMSO-d ₆ ): δ9.80 (1H, sb, NH) , 6.82-7.21 (5H, m, Ph, H-6), 6.41 (1H, m, H-1'), 4.20-4.41 (4H, H-3', H-4', H-5'), 4.01 (1H, m, Ala-NH), 3.85 (3H, s, OCH ₃ ), 3.57 (2H, m, H-α), 2, 50 (1H, m, H-2′), 2.02 (3H, s, 5-Me); ¹³ C NMR (500MHz, DMSO-d ₆ : δ173.24 (COOMe), 163.70 (C-2), 159.91 (m, Ph-para), 150.43 (C-4), 149.33 ( d, Ph-ipso, J=4Hz), 140.76 (C-6), 125.52 (m, Ph-ortho), 118.08 (m, Ph-meta), 113.52 (C-5), 84.64 (C-1′) , 82.40(C-4′)’65.87(C-5′), 60.54(C-3′), 54.78(OCH ₃ ), 50.41(C-α), 38.46(C-2′), 12.55(5- Me); ESI-MS (pos.): m/z 558 (M+H) ⁺ ; ESI-MS (neg.): m/z 556 (MH) ⁻ .

Anti-AIDS virus-1 activity experiment of the compound in CEM cells and MT-4 cells

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 9×10 ^-3 μM (CD ₅₀ 286μM)

CEM-SS 7×10 ^-2 μM (CD ₅₀ 118μM)

MT4 5×10 ^-2 μM (CD ₅₀ 60μM)

Example 12: Preparation of 3'azidovidine-5'-(p-nitrophenyl substituted)thiophosphorylalanine methyl ester compound, wherein R is CH ₃ and X is NO ₂ .

The structural formula of the compound:

The synthetic steps of compound are as follows:

1) Under the protection of nitrogen, cool in an ice-salt bath to -4～-8°C, dissolve 1mmol (0.273g) of (p-nitrobenzene substituted)phosphorous dichloride in dried tetrahydrofuran (THF), and prepare into a 1mol/L solution.

2) Add 1 mmol (0.14 g) of alanine methyl ester hydrochloride to the above solution, stir well and slowly add 2 mmol (0.2 g) of triethylamine dropwise.

3) track the reaction process with a nuclear magnetic resonance instrument (NMR), and after the (p-nitrobenzene substituted) dichloride dichloride has completely reacted, slowly dissolve 1mmol (0.267g) of 3' azidothymidine dissolved in THF Drop into the above system, stir evenly and continue to drop 1mmol (0.1g) triethylamine.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(p-nitrophenyl substituted)thiophosphorylalanine methyl ester, the yield was 71.3%.

Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ70.60, 71.30; ¹ H NMR (500 MHz, DMSO-d ₆ ): δ 9.82 (1H, sb, NH), 6.82-7.28 (5H, m, Ph, H-6), 6.36 (1H, m, H-1′), 4.20-4.41 (4H, H-3′, H-4′, H-5′), 4.11 (1H, m, Ala-NH), 3.86 (3H, s, OCH ₃ ), 3.66 (2H, m, H-α), 2, 50 (1H, m, H-2′), 2.02 (3H, s , 5-Me), 1.40 (3H, d, Ala-Me); ¹³ C NMR (500MHz, DMSO-d ₆ ): δ177.24 (COOMe), 161.70 (C-2), 159.91 (m, Ph-para ), 150.43(C-4), 149.33(d, Ph-ipso, J=4Hz), 140.76(C-6), 125.52(m, Ph-ortho), 116.08(m, Ph-meta), 111.52(C -5), 84.94(C-1′), 82.40(C-4′), 65.87(C-5′), 60.54(C-3′), 54.78(OCH ₃ ), 50.41(C-α), 35.46 (C-2'), 19.86 (d, Ala-Me, J=5Hz), 12.60 (5-Me); ESI-MS (pos.): m/z 572 (M+H) ⁺ ; ESI-MS (neg .): m/z570 MH) ^- .

Anti-AIDS virus-1 activity experiment of the compound in CEM cells and MT-4 cells

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 9×10 ^-3 μM (CD ₅₀ 168μM)

CEM-SS 6×10 ^-2 μM (CD ₅₀ 183μM)

MT4 7×10 ^-2 μM (CD ₅₀ 98μM)

Example 13: Preparation of 3'azidethymidine-5'-(p-nitrophenyl substituted)thiophosphoryl phenylalanine methyl ester compound, wherein R is C ₆ H ₅ CH ₂ , X is NO ₂ . The structural formula of the compound:

The synthetic steps of compound are as follows:

1) Under the protection of nitrogen, cool in an ice-salt bath to -4～-8C, dissolve 1mmol (0.273g) of (p-nitrobenzene substituted)phosphorous dichloride in dried tetrahydrofuran (THF), and prepare 1mol/L solution.

2) Add 1 mmol (0.215 g) of phenylalanine methyl ester hydrochloride to the above solution, stir well and slowly add 2 mmol (0.2 g) triethylamine dropwise.

3) track the reaction process with a nuclear magnetic resonance instrument (NMR), and after the (p-nitrobenzene substituted) thionyl dichloride has completely reacted, slowly dissolve 1mmol (0.266g) of 3' azidothymidine dissolved in THF Drop into the above system, stir evenly and continue to drop 1mmol (0.1g) triethylamine.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(p-nitrophenyl substituted)thiophosphoryl phenylalanine methyl ester, the yield was 61.8%. Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ71.70, 72.30; ¹ H NMR (500 MHz, DMSO-d ₆ ): δ9.81 (1H, sb, NH) , 6.82-7.25 (5H, m, Ph, H-6), 7.15-7.35 (5H, m, Ph), 6.32 (1H, m, H-1′), 4.20-4.44 (4H, H-3′, H-4', H-5'), 4.01 (1H, m, Ala-NH), 3.88 (3H, s, OCH ₃ ), 3.57 (2H, m, H-α, 2, 50 (1H, m, H-2′), 2.32 (2H, m, H-β), 2.02 (3H, s, 5-Me); ¹³ C NMR (500MHz, DMSO-d ₆ ): δ174.39 (COOMe), 163.66 (C -2), 159.91(m, Ph-para), 150.43(C-4), 149.33(d, Ph-ipso, J=4Hz), 140.76(C-6), 125.52(m, Ph-ortho), 117.08 (m, Ph-meta), 111.52 (C-5), 84.94 (C-1′), 82.40 (C-4′), 65.87 (C-5′), 60.54 (C-3′), 56.82 (C -β), 54.78 (OCH ₃ ), 50.41 (C-α), 37.46 (C-2′), 12.60 (5-Me); ESI-MS (pos.): m/z 636 (M+H) ⁺ ; ESI-MS (neg.): m/z 634 (MH) ^- the anti-AIDS virus-1 activity experiment of this compound in CEM cells and MT-4 cells

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 8×10 ^-3 μM (CD ₅₀ 185μM)

CEM-SS 6×10 ^-2 μM (CD ₅₀ 157μM)

MT4 6×10 ^-2 μM (CD ₅₀ 122μM)

Example 14: Preparation of 3'azidethymidine-5'-(p-nitrophenyl substituted)thiophosphorylvaline methyl ester compound, wherein R is (CH ₃ ) ₂ CH ₂ , and X is NO ₂ .

The structural formula of the compound:

The synthetic steps of compound are as follows:

1) Under the protection of nitrogen, cool in an ice-salt bath to -4～-8C, dissolve 1mmol (0.273g) of (p-nitrobenzene substituted)phosphorous dichloride in dried tetrahydrofuran (THF), and prepare 1mol/L solution.

2) Add 1mmol (0.17g) of valine methyl ester hydrochloride to the above solution, stir well and slowly add 2mmol (0.2g) triethylamine dropwise.

3) track the reaction process with a nuclear magnetic resonance (NMR), and after the (p-nitrobenzene substituted) thionyl dichloride has completely reacted, slowly dissolve 1mmol (0.31g) of 3' azidothymidine dissolved in THF Drop into the above system, stir evenly and continue to drop 1mmol (0.1g) triethylamine.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(p-nitrophenyl substituted)thiophosphoryl valine methyl ester, the yield was 62.3%.

Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ71.60, 72.50; ¹ H NMR (500 MHz, DMSO-d ₆ ): δ9.56 (1H, sb, NH) , 6.82-7.35 (5H, m, Ph, H-6), 6.32 (1H, m, H-1'), 4.20-4.41 (4H, H-3', H-4', H-5'), 4.01 (1H, m, Ala-NH), 3.88 (3H, s, OCH ₃ ), 3.69 (1H, m, H-α), 3.55 (1H, m, H-β), 2, 56 (1H, m , H-2'), 2.12 (3H, s, 5-Me), 1.51 (3H, s, CH ₃ ), 1.42 (3H, s, CH ₃ ); ¹³ C NMR (500MHz, DMSO-d ₆ ): δ176.24 (COOMe), 165.70 (C-2), 161.91 (m, Ph-para), 150.43 (C-4), 149.33 (d, Ph-ipso, J=4Hz), 140.76 (C-6), 125.52 (m, Ph-ortho), 117.08 (m, Ph-meta), 111.52 (C-5), 84.94 (C-1′), 82.40 (C-4′), 65.87 (C-5′), 60.54 (C-3'), 54.78(OCH ₃ ), 50.41(C-α), 48.66(C-β), 26.86(CH ₃ ), 26.40(CH ₃ ), 37.46(C-2'), 12.60(5 -Me); ESI-MS (pos.): m/z 598 (M+H) ⁺ ; ESI-MS (neg.): m/z 596 MH) ^- . Anti-AIDS virus-1 activity experiment of the compound in CEM cells and MT-4 cells

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 9×10 ^-3 μM (CD ₅₀ 178μM)

CEM-SS 5×10 ^-2 μM (CD ₅₀ 48μM)

MT4 8×10 ^-2 μM (CD ₅₀ 162μM)

Example 15: Preparation of 3'azidovidine-5'-(p-nitrophenyl substituted)thiophosphorylleucine methyl ester compound, wherein R is (CH ₃ ) ₂ CHCH ₂ , and X is NO ₂ .

The structural formula of the compound:

The synthetic steps of compound are as follows:

1) Under the protection of nitrogen, cool in an ice-salt bath to -4～-8C, dissolve 1mmol (0.273g) of (p-nitrobenzene substituted)phosphorous dichloride in dried tetrahydrofuran (THF), and prepare 1mol/L solution.

2) Add 1 mmol (0.184 g) of leucine methyl ester hydrochloride to the above solution, stir well and slowly add 2 mmol (0.2 g) of triethylamine dropwise.

3) track the reaction process with a nuclear magnetic resonance (NMR), and after the (p-nitrobenzene substituted) thionyl dichloride has completely reacted, slowly dissolve 1mmol (0.31g) of 3' azidothymidine dissolved in THF Drop into the above system, stir evenly and continue to drop 1mmol (0.1g) triethylamine.

4) After monitoring the reaction with NMR, filter, remove the solvent and other low-boiling substances by rotary distillation, carry out column chromatography separation with a silica gel column, and the eluent is chloroform:methanol=50:1 to obtain the product 3' azidothorax Glycoside-5'-(p-nitrophenyl substituted)thiophosphorylleucine methyl ester, the yield was 65.3%. Spectral data are as follows: ³¹ P NMR (DMSO-d ₆ , δ: ppm, J: Hz): δ71.8, 72.3; ¹ H NMR (500 MHz, DMSO-d ₆ ): δ9.85 (1H, sb, NH) , 6.82-7.21 (5H, m, Ph, H-6), 6.32 (1H, m, H-1′), 4.20-4.44 (4H, H-3′, H-4′, H-5′), 4.01 (1H, m, Ala-NH), 3.88 (3H, s, OCH ₃ ), 3.57 (2H, m, H-α), 3.45 (1H, m, H-β), 3.21 (1H, m, H -γ), 2, 50 (1H, m, H-2'), 2.02 (3H, s, 5-Me), 1.61 (3H, s, CH ₃ ), 1.42 (3H, s, CH ₃ ); ¹³ C NMR (500MHz, DMSO-d ₆ ): δ173.24 (COOMe), 163.70 (C-2), 159.91 (m, Ph-para), 150.43 (C-4), 149.33 (d, Ph-ipso, J =4Hz), 140.76(C-6), 125.52(m, Ph-ortho), 117.08(m, Ph-meta), 111.52(C-5), 84.94(C-1'), 82.40(C-4' ), 65.87(C-5′), 60.54(C-3′), 54.78(OCH ₃ ), 50.41(C-α), 48.66(C-β), 46.35(C-γ), 23.40(CH ₃ ) , 22.86 (CH ₃ ), 37.46 (C-2′), 12.60 (5-Me); ESI-MS (pos.): m/z612 (M+H) ⁺ ; ESI-MS (neg.): m/ z610(MH) ^- .The anti-AIDS virus-1 activity experiment of this compound in CEM cells and MT-4 cells

Aizi virus = Human immunodeficiency virus

MT-4＝Human leukenia T cell

CEM＝Human lymphoblastoid T cell

ED ₅₀ = Antiviral Activity Index

CD ₅₀ = Cytotoxicity indicator

ED ₅₀ CEM-TK ^- 8×10 ^-3 μM (CD ₅₀ 159μM)

CEM-SS 6×10 ^-2 μM (CD ₅₀ 88μM)

MT4 4×10 ^-2 μM (CD ₅₀ 121μM)

Claims (2)

1, a kind of 3 ' Zidovodine-5 '-(aromatic base 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, X is H, Cl or NO ₂In any.

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 (aromatic base 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 after stirring, acid binding agent is a triethylamine, and the add-on of acid binding agent is 2 times of amount of raw material;

3) follow the tracks of above-mentioned reaction process, after treating that the raw material total overall reaction finishes, to slowly splash in the solution in second step with 3 ' the Zidovodine solution that is dissolved in the tetrahydrofuran (THF) of the amount of raw material same substance, the acid binding agent with the amount of raw material same substance is continued to drip in the back that stirs

4) after monitoring reaction is finished, filter, rotary distillation removes and desolvates and other low-boiling point materials, carries out column chromatography for separation with silicagel column, promptly obtain 3 ' Zidovodine-5 '-(aromatic base replacement) thiophosphoryl amino acid ester compound.