CN103980238B - A benzolide compound and its application in the preparation of anti-AIDS drugs - Google Patents

Abstract

The invention belongs to medical art, specifically disclose a kind of benzo lactone compound and preparing the application in anti-AIDS drug.With styrene carboxylic acid's derivative and halogenating agent for raw material, the halogen lactonization reaction of being induced by alkali has prepared a series of benzo five-membered lactone compound and the hexa-atomic lactone compound of benzo, this benzo lactone compound and existing anti-AIDS drug efavirenz and some anti-HIV activity compound chromone analog derivatives have certain similarity, and this patent is mainly introduced structure, the preparation method of this compounds and prepared the application in anti-AIDS drug.

Description

A benzolide compound and its application in the preparation of anti-AIDS drugs

technical field

The invention belongs to the technical field of medicine, and relates to a benzolide compound and its application in preparing anti-AIDS drugs.

Background technique

AIDS (AIDS), the full name of Acquired Immunodeficiency Syndrome, is a very harmful infectious disease caused by HIV infection. The latest figures from World Guardian show that about 3 million people worldwide have died from AIDS, and there are still 30 million patients who are still struggling with AIDS. Human immunodeficiency virus type 1 (HIV-1) is the pathogenic factor causing acquired immunodeficiency syndrome. After the virus infects the host, it mainly uses reverse transcriptase, integrase and protease to complete its replication cycle. Because HIV is constantly mutating during the replication process, it has brought huge challenges to the development of drugs and vaccines. At present, the combination of several drugs with different mechanisms of action is mainly used clinically, that is, highly active antiretroviral therapy. The promotion and use of highly active antiretroviral therapy has effectively alleviated the condition of patients and prolonged the average life expectancy of AIDS patients, but the cost of treatment is high, and long-term use has produced unbearable adverse reactions and drug resistance. Although many medical researchers around the world have made great efforts, there is still no specific drug for the cure of AIDS, and there is no effective vaccine for prevention. The goal of treatment at this stage is to maximize and sustainably reduce the viral load, Obtaining immune function reconstruction and maintaining immune function, so the development of efficient and cheap therapeutic drugs is still a top priority in the prevention and treatment of AIDS. AIDS has been included in my country's Class B legal infectious diseases, and has been listed as one of the border health surveillance infectious diseases.

Reverse transcriptase (Reversetranscriptase, RT) is an important enzyme in the HIV replication process, which does not exist in human cells, but an inhibitor that inhibits this enzyme has been found in the animal research process, so that the research can be carried out Anti-AIDS drugs that target reverse transcriptase are possible, and reverse transcriptase inhibitor drugs are mainly divided into nucleosides and non-nucleosides. Nucleoside reverse transcriptase inhibitors are synthetic HIV DNA reverse transcriptase substrate deoxynucleotide analogues, which are converted into active nucleoside triphosphate derivatives in vivo and compete with natural deoxynucleoside triphosphates and HIV reverse transcriptase (RT) binds, inhibits the function of RT, and hinders the synthesis of provirus. Unlike nucleoside reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors act by approx. The hydrophobic pocket of the reverse transcriptase regulates or affects the conformation of reverse transcriptase through isomerization, destroying its activity of synthesizing DNA.

Due to the advantages of high efficiency and low toxicity, non-nucleoside reverse transcriptase inhibitors have been widely studied and clinically applied in the past 20 years, and they are an important part of high-efficiency antiretroviral therapy. At least 50 non-nucleoside reverse transcriptase inhibitors with different structures have been found. The first-generation non-nucleoside drugs are nevirapine and delavirdine, and the second-generation non-nucleoside drugs are efavirenz, etravirine, and rilpivirine . The above-mentioned non-nucleoside drugs have the advantages of strong activity and good specificity, and do not affect the DNA synthesis of cells or mitochondria, but their clinical use is limited due to the emergence of drug-resistant mutants. Therefore, there is an urgent need to develop new non-nucleoside inhibitors to broaden the options for clinical medication. The structure of flavonoids is relatively simple and has certain excellent physiological activities. It is the core skeleton structure of many drugs. Recently, it has been reported in the literature that chromones have certain anti-HIV reverse transcriptase activity, but most of them have complex structures and are difficult to synthesize. , and the activity is not particularly good, thus limiting the possibility of its medicine.

Contents of the invention

The technical problem to be solved by the present invention is to provide a benzolide compound and its application in the preparation of anti-AIDS drugs.

The benzolide compound provided by the present invention has the structure shown in the following general formula:

in,

R ¹ is H, 5-Me, 5-Cl, 4,5,6,7-Cl ₄ or 4,5,6,7-Br ₄ ;

R ² is H, Me, Et, C ₃ H ₇ , Ph,

R ³ is H, Et, C ₃ H ₇ , Ph,

Through the in vitro anti-HIV-1 activity experiment, the present invention finds that the above-mentioned benzolide compounds can be used as HIV-1 non-nucleoside reverse transcriptase inhibitors to prepare anti-AIDS drugs. Preferred, especially the following compounds:

3-(Chloro(2-chlorophenyl)methyl)isobenzofuran-1(3H)(HX1),

3-(Chloro(3-toluene)methyl)isobenzofuran-1(3H)(HX2),

3-(Chloro(3-chlorophenyl)methyl)isobenzofuran-1(3H)(HX3),

3-(Chloromethyl)-3-phenylisobenzofuran-1(3H)(HX4),

3-(Chloromethyl)-3-p-tolylisobenzofuran-1(3H)(HX5),

3-(Chloro(phenyl)methyl)isobenzofuran-1(3H)(HX6),

4-Chloro-3-(2-chlorobenzene)isochroman-1(HX7) or

4-Chloro-3-methyl-4-benzisochroman-1 (HX8).

The present invention also provides a preparation method of the benzolactone compound represented by the above structural formula.

Carboxylic acid derivatives with R ¹ , R ² , and R ³ are synthesized through the reactions shown in the following formulas i, ii, and iii. The specific operation steps can be:

Take methyl o-iodobenzoate (262mg, 1mmol), o-chlorostyrene (168mg, 1.2eq.), palladium acetate (21.6mg, 3.2%), triphenylphosphine (16.8mg, 6.4%), triethylamine (215mg, 2.13eq.) in a single-necked round bottom flask, reflux reaction at 100°C, TLC monitoring the reaction, column chromatography separation and alkaline hydrolysis to obtain the reaction product as the raw material of reaction iv.

Under the condition of anhydrous and oxygen-free, add Zn powder (392mg, 6mmol), TiCl ₄ (567.5mg, 3mmol) and THF solution, after refluxing for 2h, add carboxylic acid benzophenone (226mg, 1mmol) and acetaldehyde (220mg , 5mmol) THF solution, continue to reflux reaction overnight, TLC monitors the reaction, column chromatography separates and obtains the product of the reaction as the raw material of the reaction iv.

Under anhydrous and oxygen-free conditions, add methyltriphenylphosphine bromide (571.6mg, 1.6mmol), THF solution, add potassium tert-butoxide (264.6mg, 2.236mmol) under ice bath, stir for 30min, then add carboxylic acid Benzophenone (226 mg, 1 mmol) was reacted at room temperature, monitored by TLC, and separated by column chromatography to obtain a reaction product as a raw material for reaction iv.

Through the synthesis of commercial products, obtain carboxylic acid derivatives with R ¹ , R ² , R ³ , then take equivalent amounts of carboxylic acid derivatives with R ¹ , R ² , R ³ and DCDMH and dissolve them in dichloromethane to fully dissolve , and then add an appropriate amount of DABCO to catalyze the halide lactonization reaction, and monitor the progress of the reaction by TLC. After the basic reaction of the raw materials is complete, the target compound is separated by column chromatography. The reaction formula is shown in formula iv below.

in,

R ¹ is H, 5-Me, 5-Cl, 4,5,6,7-Cl ₄ ,4,5,6,7-Br ₄ etc;

R ² is H, Me, Et, C ₃ H ₇ , Ph,

R ³ is H, Et, C ₃ H ₇ , Ph,

The specific operation process can be as follows:

Take 0.2mmol of carboxylic acid derivatives of reaction i or reaction ii or reaction iii and 0.24mmol (1.2eq.) of 1,3-dichloro-5,5-dimethylhydantoin (DCDMH) into a magnetic In a single-necked round bottom flask, add 5 mL of dichloromethane to dissolve it, stir evenly, add 0.02 mmol of triethylenediamine (DABCO) to catalyze the halide lactonization reaction, and monitor the reaction by TLC. After the basic reaction of the raw materials is complete, pure benzolide compounds are obtained through column chromatography separation.

Under anhydrous and oxygen-free conditions, benzolactone (35.2mg, 0.1mmol), PdCl ₂ (PPh ₃ ) ₂ (3.5mg, 0.005mmol, 5%), PPh ₃ (7.9mg, 0.03mmol, 30 %), CuI (2mg, 0.01mmol, 10%), and phenylacetylene (0.16mmol) in a mixed solvent of 1mLDMF/1.5mLTEA reacted at 80°C for 24h. TLC spot plate monitoring, distilled off the solvent, and separated by column chromatography to obtain iso Shaman derivatives. The reaction formula is as follows:

The compound described in the invention can be used as an HIV-1 non-nucleoside reverse transcriptase inhibitor for the preparation of anti-AIDS drugs. Therefore, the present invention also provides an anti-AIDS pharmaceutical composition, comprising the compound described in the present invention and one or more pharmaceutically acceptable carriers or excipients, which can be prepared according to existing conventional medical techniques.

detailed description

Example 1: Preparation of 3-(chloro(2-chlorophenyl)methyl)-1(3H)-isobenzofuranone (HX1)

Take 0.2mmol of the carboxylic acid derivative of reaction i and 0.24mmol (1.2eq.) of 1,3-dichloro-5,5-dimethylhydantoin (DCDMH) into a single-necked round-bottomed flask containing a magnet Add 5mL of dichloromethane to dissolve it, stir evenly, add 0.02mmol of triethylenediamine (DABCO) to catalyze the halide lactonization reaction, and monitor the reaction by TLC. After the basic reaction of the raw materials was completed, the pure solid compound HX1 was obtained by column chromatography separation, and the product was a yellow solid with a yield of 71%. ¹ HNMR (400MHz, CDCl ₃ ) δ8.17(d, J=7.7Hz, 1H), 7.68(d, J=7.2Hz, 1H), 7.61(d, J=7.6Hz, 1H), 7.55(d, J＝7.6Hz, 1H), 7.36(t, J＝7.1Hz, 4H), 5.62(d, J＝7.5Hz, 1H), 5.33(d, J＝7.5Hz, 1H). ¹³ CNMR (101MHz, CDCl ₃ ) δ163.16, 137.65, 135.28, 134.83, 134.29, 130.55, 129.79, 129.20, 128.99, 128.49, 127.50, 127.11, 123.79, 83.36, 56.27.

Example 2: Preparation of 3-(chloro(3-toluene)methyl)-1(3H)-isobenzofuranone (HX2)

The preparation method was as in Example 1, and the product was a white solid with a yield of 83%. ¹ HNMR (400MHz, CDCl ₃ ) δ8.17(d, J=7.7Hz, 1H), 7.64(d, J=7.4Hz, 1H), 7.58(d, J=7.6Hz, 1H), 7.51(t, J＝7.5Hz, 1H), 7.25(d, J＝6.2Hz, 1H), 7.15(dd, J＝18.1, 7.5Hz, 3H), 5.65(d, J＝6.7Hz, 1H), 5.39(d, J＝6.7Hz,1H),2.34(s,3H) ^.13 CNMR(101MHz,CDCl ₃ )δ163.38,138.59,137.79,135.74,134.65,130.44,130.01,129.70,128.60,127.60,127.56,124.3135,56 ,21.47.

Example 3: Preparation of 3-(chloro(3-chlorophenyl)methyl)-1(3H)-isobenzofuranone (HX3)

The preparation method was as in Example 1, and the product was a white solid with a yield of 87%. ¹ HNMR (400MHz, CDCl ₃ ) δ8.17(d, J=7.7Hz, 1H), 7.70–7.66(m, 1H), 7.61(d, J=7.5Hz, 1H), 7.54(t, J=7.5 Hz,1H),7.38–7.32(m,4H),5.62(d,J＝7.5Hz,1H),5.33(d,J＝7.5Hz,1H). ¹³ CNMR(101MHz,CDCl ₃ )δ163.15,137.66, 135.29, 134.82, 134.29, 130.56, 129.79, 129.20, 128.99, 128.49, 127.49, 127.11, 123.80, 83.36, 56.27.

Example 4: Preparation of 3-(chloromethyl)-3-phenyl-1(3H)-isobenzofuranone (HX4)

The preparation method is as in Example 1, and the product is a white solid with a yield of 80%. ¹ HNMR (400MHz, CDCl ₃ ) δ7.86(d, J=7.6Hz, 1H), 7.70–7.63(m, 1H), 7.60(d, J=7.7Hz, 1H), 7.55–7.45(m, 3H ),7.37–7.25(m,3H),4.20–4.09(m,2H). ¹³ CNMR(101MHz,CDCl ₃ )δ164.75,144.94,132.93,130.11,125.70,124.91,124.72,122.30,121.75,121.249,118 83.64, 45.15.

Example 5: Preparation of 3-(chloromethyl)-3-p-tolyl-1(3H)-isobenzofuranone (HX5)

The preparation method was as in Example 1, and the product was a light yellow solid with a yield of 82%. ¹ HNMR (400MHz, CDCl ₃ ) δ7.93(d, J=7.6Hz, 1H), 7.76–7.71(m, 1H), 7.66(d, J=7.7Hz, 1H), 7.61–7.55(m, 1H ), 7.43(d, J＝8.3Hz, 2H), 7.20(d, J＝8.1Hz, 2H), 4.25–4.17(m, 2H), 2.34(s, 3H). ¹³ CNMR(101MHz, CDCl ₃ ) δ164.84, 145.11, 134.94, 130.06, 129.94, 125.61, 125.37, 122.33, 121.69, 121.21, 118.48, 83.69, 45.14, 16.79.

Example 6: Preparation of 3-(chloro(phenyl)methyl)-1(3H)-isobenzofuranone (HX6)

The preparation method was as in Example 1, and the product was a white solid with a yield of 81%. ¹ HNMR (400MHz, CDCl ₃ )δ7.89–7.79(m,1H),7.54(s,2H),7.41(s,2H),7.36(d,J＝16.5Hz,4H),5.84(t,J ＝5.5Hz, 1H), 5.21(dd, J＝18.6, 5.6Hz, 1H). ¹³ CNMR (101MHz, CDCl ₃ ) δ169.34, 145.78, 135.55, 133.82, 129.97, 129.13, 128.68, 128.57, 128.38, 128.077, 125. ,123.75,123.29,82.60,62.77.

Example 7: Preparation of 4-chloro-3-(2-chlorobenzene)-1-isochromanone (HX7)

The preparation method was as in Example 1, and the product was a light yellow solid with a yield of 21%. ¹ HNMR (400MHz, CDCl ₃ ) δ7.97–7.82(m,2H),7.74–7.69(m,1H),7.63–7.56(m,2H),7.37(dt,J＝12.6,6.4Hz,3H) ,5.93–5.82(m,1H),4.55(dd,J＝54.7,11.9Hz,1H) ^.13 CNMR(101MHz,CDCl ₃ )δ170.39,134.33,133.74,132.51,130.65,130.36,130.22,130.06,129.81, 127.32, 125.80, 123.92, 123.62, 81.03, 58.62.

Example 8: Preparation of 3-(1-chloroethyl)-3-phenyl-1(3H)-isobenzofuranone (HX8)

The preparation method was as in Example 1, and the product was a light yellow solid with a yield of 69%. ¹ HNMR (400MHz, CDCl ₃ ) δ7.91(d, J=7.6Hz, 1H), 7.79(d, J=7.8Hz, 1H), 7.72(t, J=7.4Hz, 1H), 7.63–7.51( m, 3H), 7.38(dt, J＝21.9, 7.1Hz, 3H), 4.88(q, J＝6.6Hz, 1H), 1.46(d, J＝6.7Hz, 3H). ¹³ CNMR (101MHz, CDCl ₃ )δ169.37, 151.89, 150.31, 134.29, 129.74, 128.97, 128.79, 125.91, 125.20, 122.71, 90.31, 61.18, 19.83.

Example 9: Preparation of 3-(chloromethyl)-3-p-fluorophenyl-1(3H)-isobenzofuranone (HX9)

The preparation method is as in Example 1, and the product is a light yellow solid with a yield of 80%. ¹ HNMR (400MHz, CDCl ₃ ) δ7.95(d, J=7.6Hz, 1H), 7.77(t, J=7.5Hz, 1H), 7.68(d, J=7.7Hz, 1H), 7.62(t, J=7.5Hz,1H),7.58–7.48(m,2H),7.10(dd,J=11.9,5.3Hz,2H),4.23–4.16(m,2H). ¹³ CNMR(101MHz,CDCl ₃ )δ168. 82,163.00(d, ¹ J _CF ＝250.48Hz),148.99,134.52,133.06,133.03,130.17,127.75(d, ² J _CF ＝9.09Hz),126.54,126.20,122.81,116.00(d, ³ J _2Hz2 ＝22. ), 115.89, 87.50, 77.39, 77.07, 76.75, 49.39.

Example 10: Preparation of 3-(1-chloroethyl)-3-p-fluorophenyl-1(3H)-isobenzofuranone (HX10)

The preparation method was as in Example 1, and the product was a light yellow solid with a yield of 81%. ¹ HNMR (400MHz, CDCl ₃ ) δ7.94 (d, J = 7.3Hz, 1H), 7.78 (dd, J = 15.1, 7.3Hz, 2H), 7.63 (dd, J = 12.7, 7.6Hz, 3H), 7.09(t, J=8.4Hz, 2H), 4.69(q, J=6.4Hz, 1H), 1.44(d, J=6.6Hz, 3H). ¹³ CNMR(101MHz, CDCl ₃ )δ171.51, 162.80(d, ¹ J _CF ＝249.47Hz),148.98,134.29,130.10,128.00(d, ² J _CF ＝8.08Hz),127.79,127.70,126.45,126.37,123.51,120.69,115.66(d, ³ J _CF ＝22.22Hz), 90.08, 61.19, 19.74.

Example 11: Preparation of 3-(1-chloroethyl)-3-p-chlorophenyl-1(3H)-isobenzofuranone (HX11)

The preparation method was as in Example 1, and the product was a yellow solid with a yield of 82%. ¹ HNMR (400MHz, CDCl ₃ ) δ7.93(d, J=7.7Hz, 1H), 7.80(d, J=7.6Hz, 1H), 7.75(d, J=7.4Hz, 1H), 7.65(d, J＝7.4Hz, 1H), 7.60(d, J＝7.3Hz, 1H), 7.42–7.35(m, 3H), 4.77(q, J＝6.4Hz, 1H), 1.44(d, J＝6.9Hz, 3H). ¹³ CNMR (101MHz, CDCl ₃ ) δ169.08, 149.07, 137.35, 134.17, 129.97, 128.98, 128.83, 128.71, 126.60, 126.25, 125.93, 125.20, 123.55, 90.61, 691.23,

Example 12: Preparation of 5-chloro-3-(1-chloroethyl)-3-p-fluorophenyl-1(3H)-isobenzofuranone (HX12)

The preparation method was as in Example 1, and the product was light yellow oil with a yield of 85%. ¹ HNMR (400MHz, CDCl ₃ ) δ7.94 (d, J = 7.5Hz, 1H), 7.81–7.74 (m, 2H), 7.64 (dd, J = 5.5, 3.5Hz, 2H), 7.08 (dd, J =11.8,5.4Hz,2H),4.69(q,J=6.7Hz,1H),1.44(d,J=6.8Hz,3H). ¹³ CNMR(101MHz,CDCl ₃ )δ168.83,162.80(d, ¹ J _CF ＝249.47Hz),148.98,134.30,133.15,130.10,128.00(d, ² J _CF ＝9.09Hz),127.78,126.45,126.35,126.21,123.50,115.66(d, ³ J _CF ＝21.21Hz),90.09, ,19.74.

Example 13: Preparation of 5-methyl-3-(1-chloroethyl)-3-p-fluorophenyl-1(3H)-isobenzofuranone (HX13)

The preparation method is as in Example 1, and the product is a white solid with a yield of 80%. ¹ HNMR (400MHz, CDCl ₃ ) δ8.07 (d, J = 8.0Hz, 1H), 7.42 (dd, J = 8.7, 5.3Hz, 2H), 7.29 (d, J = 7.9Hz, 1H), 7.10 ( t, J=8.7Hz, 2H), 6.73(s, 1H), 4.87(q, J=6.5Hz, 1H), 2.32(s, 3H), 1.23(d, J=6.5Hz, 3H). ¹³ CNMR (101MHz,CDCl ₃ )δ164.77,160.43(d, ¹ J _CF ＝148.47Hz),145.78,144.78,136.15,130.47(d, ² J _CF ＝24.24Hz),128.63,128.55,127.77,121.61,115.31(d, ³ J _CF ＝21.21Hz), 81.42, 73.74, 21.94, 13.95.

Example 14: Preparation of 3-(1-iodoethyl)-3-p-fluorophenyl-1(3H)-isobenzofuranone (HX14)

The preparation method was as in Example 1, and the product was a yellow solid with a yield of 78%. ¹ HNMR (400MHz, CDCl ₃ ) δ7.93(d, J=7.6Hz, 1H), 7.74(t, J=7.5Hz, 1H), 7.66–7.58(m, 2H), 7.54(dd, J=8.9 ,5.1Hz,2H),7.07(t,J=8.6Hz,2H),4.69(dd,J=11.5,2.1Hz,1H),1.79(ddd,J=14.5,11.5,7.2Hz,1H),1.34 (ddd, J＝14.5, 7.2, 2.1Hz, 1H), 1.01 (t, J＝7.1Hz, 3H). ¹³ CNMR (101MHz, CDCl ₃ ) δ168.50, 162.71 (d, ¹ J _CF ＝249.47Hz), 149.07 ,138.81,135.36,134.35,130.03,127.31(d, ² J _CF ＝8.08Hz),127.09,126.39,122.74,115.82(d, ³ J _CF ＝22.22Hz),90.66,44.75,27.76,15.08.

Example 15: Preparation of 3-(1-chlorobutyl)-3-p-fluorophenyl-1(3H)-isobenzofuranone (HX15)

The preparation method was as in Example 1, and the product was a light yellow solid with a yield of 76%. ¹ HNMR (400MHz, CDCl ₃ ) δ7.93 (t, J = 8.0Hz, 1H), 7.78 (ddd, J = 15.6, 11.2, 6.0Hz, 2H), 7.66–7.54 (m, 3H), 7.13–7.05 (m,2H),4.56(ddd,J=65.2,10.9,2.2Hz,1H),1.64(tt,J=11.8,7.5Hz,3H),1.36(ddd,J=17.1,11.5,6.2Hz,1H ),0.84(t,J＝7.1Hz,3H). ¹³ CNMR(101MHz,CDCl ₃ )δ168.82,149.46(d, ¹ J _CF ＝86.86Hz),134.36,134.20,130.03,129.86,128.08(d, ² J _CF ＝8.08Hz),128.05,127.44,126.30,126.04,123.78,116.01,115.65(d, ³ J _CF ＝21.21Hz),90.18,66.76,34.42,19.96,13.23.

Example 16: Preparation of 4-chloro-3-methyl-4-(4-phenylethynylphenyl)-1-isochromanone (HX16)

The preparation method is as in Example 1, the product is light yellow solid, and the yield is 95%. ¹ HNMR (400MHz, CDCl ₃ ): d=7.92(t, J=7.1Hz, 1H), 7.79(d, J=7.7Hz, 1H), 7.74(t, J=7.7Hz, 1H), 7.57(q ,J=8.6Hz,5H),7.52(dd,J=6.5,3.0Hz,2H),7.41–7.31(m,3H),4.86(q,J=6.7Hz,1H),1.46(d,J= 6.6Hz,3H)； ¹³ CNMR(101MHz,CDCl ₃ ):d＝169.19,149.89,137.99,134.44,132.10,131.67,129.91,129.74,128.99,128.59,128.42,126.20,126.02,125.90,125.36,125.20,124.04 ,122.86,122.73,90.70,90.11,88.37,60.98,19.81.

The chemical structure of the target compound HX1-16 synthesized by the method of the present invention in table 1

^a Replace DCDMH with NIS

Experimental Example 22: Pharmacological experiment of benzolide compounds

(1) Determination of Cytotoxicity of Benzolide Compounds:

Yellow thiazolan, referred to as MTT, can enter the cell through the cell membrane, and the amber dehydrogenase in the mitochondria of living cells can reduce the exogenous MTT to blue-purple needle-shaped Formazan crystals that are insoluble in water and deposit in the cell. The crystals can be dissolved by 20% (mass to volume) SDS, and the light absorption value is measured at a wavelength of 595nm by an enzyme-linked immunosorbent detector, which can indirectly reflect the number of cells.

During the experiment, TZM-bl (provided by the National Institutes of Health) cells were transferred to a 96-well plate, and the compound was added to the cells at a certain dilution after 24 hours. After culturing at 37°C for 72 hours, 100 μL of the supernatant was sucked away, and 20 μl of MTT was added. After continuing to culture at 37° C. for 4 hours, 100 μL of 20% SDS was added and cultured for 18 hours, and the OD value at a wavelength of 595 nm was measured with a microplate reader. Inhibition rate of the compound (%)=[1-(EN)/(PN)]×100, where "E" represents the OD value of the administration group, "P" represents the OD value of the non-administration group, and "N" represents OD value of blank group. The half inhibitory concentration (CC ₅₀ ) of a compound was used as an indicator of the cytotoxicity of the compound.

(2) Anti-HIV-1 activity of benzolide compounds in vitro:

HIV _IIIB (provided by the National Institutes of Health) is a classic HIV drug screening experimental strain, TZM-bl (provided by the National Institutes of Health) cells are modified Hela (provided by the American Type Collection) cell line, and the cell membrane stably expresses HIV receptor and coreceptor, nuclei stably integrate the HIV long terminal repeat-initiated luciferase gene. After HIV-ⅢB infects TZM-b1 cells, the TAT protein expressed by the virus can activate the expression of luciferase gene in the cells, and the level of virus replication can be judged by detecting the activity of luciferase in the cells. By detecting the luciferase in the cells after drug treatment, the activity of the drug to inhibit the HIV-1 virus can be accurately quantified.

During the experiment, the HIV _IIIB virus was mixed with the drug and added to the TZM-bl cells (60 (area ratio) was confluent). After the virus was adsorbed to the cells for 2 hours, the mixture of the virus and the drug was sucked off, and fresh medium (DMEM, 90% (volume ratio), fetal bovine serum, 10% (volume ratio), G418, 500 μg/ml; hygromycin, 100 μg/mL; puromycin, 1 μg/mL. The percentage content is the volume percentage content ) to continue culturing, and do 8 replicate wells for each dilution. After incubation at 37°C for 24 hours, the luciferase activity in the cells was detected. Compound inhibition rate (%)=[1-(EN)/(PN)]×100, where "E" represents the activity of luciferase in the experimental group, "P" represents the activity of luciferase in the positive group, "N" represents the luciferase activity in the negative group. The half inhibitory concentration (IC ₅₀ ) of the compound was used as an indicator of its antiviral activity.

Cytotoxicity and anti-HIV-1 activity results of the target compound HX1-16 synthesized by the present invention

Compounds _IC50 (μM) _CC50 (μM) SI (CC ₅₀ /IC ₅₀ ) HX1 0.450 136.451 301 HX2 12.468 293.333 23.5 HX3 14.452 272.903 18.9 HX4 15.462 293.333 19 HX5 47.666 293.333 6.2 HX6 30.924 293.333 9.5 HX7 12.281 136.451 11.1 HX8 29.336 293.333 10 HX9 326 290 0.9 HX10 293 276 1 HX11 201 201 1 HX12 309 247 0.8 HX13 263 263 1 HX14 126 202 1.6 HX15 314 252 0.8 HX16 242 215 0.9

The above experimental results show that most of the synthesized compounds have good anti-HIV-1 activity, such as compounds HX1 (IC ₅₀ =0.45μM, SI=301), HX2 (IC ₅₀ =12.468μM, SI=23.5), HX3 ( IC ₅₀ =14.452 μM, SI=18.9), HX4 (IC ₅₀ =15.462 μM, SI=19), HX7 (IC ₅₀ =12.281 μM, SI=11.1), etc.

Claims (3)

CLAIMS 1. A compound which is 3-(chloro(2-chlorophenyl)methyl)-1(3H)-isobenzofuranone. 2. The application of the compound described in claim 1 in the preparation of anti-AIDS drugs. 3. An anti-AIDS pharmaceutical composition, comprising the compound of claim 1 and one or more pharmaceutically acceptable carriers or excipients.