CN109810044B - A compound with HIV-1 integrase inhibitory activity and its preparation and application - Google Patents

Abstract

The invention relates to the technical field of medicines, in particular to a compound with HIV-1 integrase inhibitory activity and a preparation method thereof. The invention takes p-methylaniline as raw material, and is synthesized through a series of steps, finally a compound which has obvious antiviral activity and less influence on normal cells and has HIV-1 integrase inhibitory activity is obtained, and the compound is represented by the following general formula (I):

in the general formula (I), R1 and R2 are hydroxyl, alkoxy, amino and hydrogen atoms; r3 is hydrogen atom, halogen atom, alkyl, nitro, hydroxyl or amino.

Description

A compound with HIV-1 integrase inhibitory activity and its preparation and application

technical field

The invention relates to the technical field of medicine, in particular to a compound with HIV-1 integrase inhibitory activity and its preparation and application.

Background technique

AIDS, also known as acquired immune deficiency syndrome (AIDS), is an infectious disease characterized by severe damage to the systemic immune system caused by human immune deficiency virus (HIV) infection. Since the first case of AIDS was reported in the United States in 1981, it has continued to spread around the world and raged on a global scale, causing serious harm to human life, health and social development. HIV-1 drug resistance is due to the mutation of the viral gene, which changes the biochemical or structural characteristics of the drug target, resulting in the insensitivity or decreased sensitivity of the virus to the drug.

HIV-1 integrase (Integrase) is one of the three key enzymes in the viral replication process, which can mediate the integration of retroviral DNA into the host genomic DNA. If the integration process is blocked, the replication of the virus will be interrupted. Therefore, the integration process is the last key step for HIV-1 virus to invade target cells and eventually form an irreversible infection.

However, the integrase strand transfer inhibitors currently on the market have a problem of drug resistance. Therefore, it is an urgent problem to find an HIV-1 integrase inhibitor with novel structure, low toxicity and high antiviral activity.

SUMMARY OF THE INVENTION

In order to solve the above-mentioned technical problems existing in the prior art, the present invention provides a compound with HIV-1 integrase inhibitory activity, as follows:

A compound with HIV-1 integrase inhibitory activity, which is represented by the following general formula (I):

Preferably, in the general formula (I), R1 and R2 are hydroxyl, alkoxy, amino, and hydrogen atoms. The compound at this time has good antiviral activity.

Preferably, in the general formula (I), R3 is a hydrogen atom, a halogen atom, an alkyl group, a nitro group, a hydroxyl group, or an amino group. The compound at this time has good antiviral activity.

Further preferably, in the general formula (I), R1 is OH, R2 is OCH ₂ CH ₃ , and R3 is H. The compound at this time has better antiviral activity and lower cytotoxicity, and the overall effect is better.

The preparation method of the compound with HIV-1 integrase inhibitory activity comprises the following steps:

(1) Synthesis of p-methoxydiazobenzene hydrochloride (compound b): Dissolve 1-fold equivalent of compound a (p-toluidine) with concentrated hydrochloric acid, stir at room temperature for 2 hours; In the reaction system, when the temperature is constant, slowly add 2 times the equivalent of sodium nitrite, stir the reaction, and react at this temperature for 2-5h to obtain compound b;

(2) Synthesis of (Z)-2-acetyl-2-((4-methoxyphenyl)diazenyl)diethyl glutarate (compound c): completely dissolve 1.2 times equivalent in absolute ethanol The raw material 2-acetylglutaric acid diethyl ester was added with excess concentrated H ₂ SO ₄ to make it fully mixed, and compound b was added slowly and at a constant rate, and the reaction was carried out at room temperature for 2-3 hours. to weak basicity, extract with ethyl acetate (EthylAcetate, EA), concentrate the extract, and separate and purify the target product by silica gel column chromatography to obtain compound c;

(3) Synthesis of (Z)-2-(2-(4-methoxyphenyl)hydrazono)diethyl glutarate (compound d): After the isolated compound c was completely dissolved in absolute ethanol , add excess concentrated HCl, stir at room temperature for 2-5h, after the reaction is over, separate and purify compound d;

(4) Synthesis of 3-(2-ethoxy-2-oxoethyl)-5-methoxy-1H-indole-2-carboxylic acid ethyl ester (compound e): compound d was treated with absolute ethanol After complete dissolution, dry HCl gas was introduced, and the mixture was stirred at room temperature for 2-7 hours. After the reaction was completed, experimental post-treatment was performed, and compound e was isolated and purified, which is a compound with HIV-1 integrase inhibitory activity.

Preferably, in the reaction process of the steps (1)-(4), thin layer chromatography (Thin Layer Chromatography, TLC) is performed to monitor the reaction progress, and if the reaction is not complete, the reaction is continued until the reaction is complete. TLC is a commonly used method in chemical synthesis, and its advantages are that it is simple, fast, sensitive, and can be monitored in real time.

Preferably, the post-experimental treatment includes operations such as adjusting the pH of the reaction solution, extracting with ethyl acetate (Ethyl Acetate, EA), concentrating, and separating. Further preferably, described adjusting the pH of the reaction solution, extracting with EA, concentrating, separating and other operations, specifically adjusting the pH to weakly alkaline, extracting with EA, concentrating the extract, packing a column, and separating and purifying the target product with thin-layer chromatography.

Preferably, the described pH adjustment is weakly alkaline, and is adjusted with anhydrous sodium carbonate.

The application of the compound with HIV-1 integrase inhibitory activity in the preparation of anti-HIV medicines. In particular, the compound is used for HIV-1 type integrase inhibitor.

Compared with the prior art, the technical effect created by the present invention is embodied in:

The present invention finds that the compound containing the parent nucleus of 2-indolecarboxylic acid has not been reported on HIV-1 integrase inhibitors in the present invention through preliminary research. At the same time, it has the most significant antiviral activity, and indole is a very important type of structure, which can imitate the structure of the peptide and bind to the target protein, making it easier to reach the integrase active region, thereby enhancing its antiviral effect. Therefore, the inventors believe that 2-indolecarboxylic acid new integrase inhibitors may be more easily combined with the active site region of integrase to achieve a more significant antiviral effect. Synthesized, and finally obtained a compound with HIV-1 integrase inhibitory activity and its derivatives, which has significant antiviral activity and little impact on normal cells. , The reaction conditions are simple and easy, and the yield is high.

Description of drawings

1 is a schematic diagram of the synthesis process of compounds with HIV-1 integrase inhibitory activity and derivatives thereof;

Fig. 2 is the structural representation of the compound with HIV-1 integrase inhibitory activity of the present invention;

Detailed ways

The technical solutions of the present invention will be further limited below in conjunction with specific embodiments, but the scope of protection is not limited to the description.

Example 1

(1) Synthesis of p-methoxydiazobenzene hydrochloride (compound b): 1.00 g of compound a (p-toluidine) was completely dissolved in concentrated hydrochloric acid, stirred at room temperature for 2 hours; moved into a reaction system from 0°C to -10°C , when the temperature is constant, slowly add 0.8404g of sodium nitrite, stir the reaction, and react at this temperature for 3.5h to obtain compound b

(2) Synthesis of diethyl (Z)-2-acetyl-2-((4-methoxyphenyl)diazenyl)glutarate (compound c): 2.24 g of 2.24 g was completely dissolved in absolute ethanol - Add excess concentrated H ₂ SO ₄ to diethyl acetylglutarate to make it fully mixed, slowly and at a constant speed, add 1.7 g of ethanol solution of compound b dropwise, and react at room temperature for 2.5 hours after the completion of the dropwise addition. After-treatment, i.e. separation and purification to obtain compound c;

(3) Synthesis of (Z)-diethyl 2-(2-(4-methoxyphenyl)hydrazono)glutarate (compound d): After 1 g of compound c was completely dissolved in absolute ethanol, add Excess concentrated HCl was stirred at room temperature for 3.5 h, after the reaction was completed, experimental post-treatment was performed, and then compound d was isolated and purified;

(4) Synthesis of 3-(2-ethoxy-2-oxoethyl)-5-methoxy-1H-indole-2-carboxylic acid ethyl ester (compound e): 1 g of compound d was treated with anhydrous After the ethanol was completely dissolved, dry HCl gas was introduced, and the mixture was stirred at room temperature for 4.5 hours. After the reaction was completed, the experimental post-treatment was performed, and then compound e was isolated and purified, which was compound e with HIV-1 integrase inhibitory activity.

In the reaction process of the steps (1)-(4), thin layer chromatography (Thin Layer Chromatography, TLC) is performed to monitor the progress of the reaction, and if the reaction is not complete, the reaction is continued until the reaction is complete.

The described experimental post-treatment is to adjust the pH to weak alkaline, extract three times with ethyl acetate (EthylAcetate, EA), concentrate the extract, pack into a column, and separate and purify the target product by thin layer chromatography.

The described pH adjustment is weakly alkaline, and is adjusted with anhydrous sodium carbonate.

Compound e with HIV-1 integrase inhibitory activity

Example 2

(1) Synthesis of p-methoxydiazobenzene hydrochloride (compound b): 1.00 g of compound a (p-toluidine) was completely dissolved in concentrated hydrochloric acid, stirred at room temperature for 2 hours; moved into a reaction system from 0°C to -10°C , when the temperature is constant, slowly add 0.8404g of sodium nitrite, stir the reaction, and react at this temperature for 2h to obtain compound b

(2) Synthesis of diethyl (Z)-2-acetyl-2-((4-methoxyphenyl)diazenyl)glutarate (compound c): 2.24 g of 2.24 g was completely dissolved in absolute ethanol - Add excess concentrated H ₂ SO ₄ to diethyl acetylglutarate to make it fully mixed, slowly and at a constant speed, add 1.7g of ethanol solution of compound b dropwise, and react at room temperature for 2 hours after the completion of the dropwise addition. Treatment, i.e. separation and purification, obtains compound c;

(3) Synthesis of (Z)-diethyl 2-(2-(4-methoxyphenyl)hydrazono)glutarate (compound d): After 1 g of compound c was completely dissolved in absolute ethanol, add Excess concentrated HCl was stirred at room temperature for 2 h, after the reaction was completed, the experimental post-treatment was performed, and then compound d was isolated and purified;

(4) Synthesis of 3-(2-ethoxy-2-oxoethyl)-5-methoxy-1H-indole-2-carboxylic acid ethyl ester (compound e): 1 g of compound d was treated with anhydrous After the ethanol was completely dissolved, dry HCl gas was introduced, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the experimental post-treatment was performed, and then compound e was isolated and purified, which was compound e with HIV-1 integrase inhibitory activity.

In the reaction process of the steps (1)-(4), thin layer chromatography (Thin Layer Chromatography, TLC) is performed to monitor the progress of the reaction, and if the reaction is not complete, the reaction is continued until the reaction is complete.

The described experimental post-treatment is to adjust the pH to weak alkaline, extract three times with ethyl acetate (EthylAcetate, EA), concentrate the extract, pack into a column, and separate and purify the target product by thin layer chromatography.

The described pH adjustment is weakly alkaline, and is adjusted with anhydrous sodium carbonate.

Compound e with HIV-1 integrase inhibitory activity

Example 3

(1) Synthesis of p-methoxydiazobenzene hydrochloride (compound b): 1.00 g of compound a (p-toluidine) was completely dissolved in concentrated hydrochloric acid, stirred at room temperature for 2 hours; moved into a reaction system from 0°C to -10°C , when the temperature is constant, slowly add 0.8404g of sodium nitrite, stir the reaction, and react at this temperature for 5h to obtain compound b

(2) Synthesis of diethyl (Z)-2-acetyl-2-((4-methoxyphenyl)diazenyl)glutarate (compound c): 2.24 g of 2.24 g was completely dissolved in absolute ethanol - Add excess concentrated H ₂ SO ₄ to diethyl acetylglutarate to make it fully mixed, slowly and constant speed dropwise add 1.7g of the ethanol solution of compound b, and react at room temperature for 3h after the completion of the dropwise addition. Treatment, i.e. separation and purification, obtains compound c;

(3) Synthesis of (Z)-diethyl 2-(2-(4-methoxyphenyl)hydrazono)glutarate (compound d): After 1 g of compound c was completely dissolved in absolute ethanol, add Excess concentrated HCl was stirred at room temperature for 5h, after the reaction was completed, the experimental post-treatment was performed, and then compound d was isolated and purified;

(4) Synthesis of 3-(2-ethoxy-2-oxoethyl)-5-methoxy-1H-indole-2-carboxylic acid ethyl ester (compound e): 1 g of compound d was treated with anhydrous After the ethanol was completely dissolved, dry HCl gas was introduced, and the mixture was stirred at room temperature for 7 hours. After the reaction was completed, the experimental post-treatment was performed, and then compound e was isolated and purified, which was compound e with HIV-1 integrase inhibitory activity.

In the reaction process of the steps (1)-(4), thin layer chromatography (Thin Layer Chromatography, TLC) is performed to monitor the progress of the reaction, and if the reaction is not complete, the reaction is continued until the reaction is complete.

The described experimental post-treatment is to adjust the pH to weak alkaline, extract three times with ethyl acetate (EthylAcetate, EA), concentrate the extract, pack into a column, and separate and purify the target product by thin layer chromatography.

The described pH adjustment is weakly alkaline, and is adjusted with anhydrous sodium carbonate.

Compound e with HIV-1 integrase inhibitory activity

Example 4

(1) Synthesis of p-methoxy diazobenzene hydrochloride (compound b): Completely dissolve 1.00 g (1.00 eq.) of compound a (p-toluidine) with concentrated hydrochloric acid, stir at room temperature for 2 h; In the -10℃ reaction system, when the temperature is constant, slowly add 0.8404g of sodium nitrite, stir the reaction, and react at this temperature for 3.5h to obtain compound b

(2) Synthesis of diethyl (Z)-2-acetyl-2-((4-methoxyphenyl)diazenyl)glutarate (compound c): 2.24 g of 2.24 g was completely dissolved in absolute ethanol -Add excess concentrated H ₂ SO ₄ to diethyl acetylglutarate to make it fully mixed, slowly and steadily dropwise add the solution of compound b, and react at room temperature for 2.5 hours after the dropwise addition. That is, compound c is obtained by separation and purification;

(3) Synthesis of (Z)-diethyl 2-(2-(4-methoxyphenyl)hydrazono)glutarate (compound d): After 1 g of compound c was completely dissolved in absolute ethanol, add Excess concentrated HCl was stirred at room temperature for 3.5 h, after the reaction was completed, experimental post-treatment was performed, and then compound d was isolated and purified;

(4) Synthesis of 3-(2-ethoxy-2-oxoethyl)-5-methoxy-1H-indole-2-carboxylic acid ethyl ester (compound e): 1 g of compound d was treated with anhydrous After the ethanol is completely dissolved, dry HCl gas is introduced, and the mixture is stirred at room temperature for 4.5 hours. After the reaction is completed, the experimental post-treatment is performed, and then compound e is isolated and purified, which is the compound e with HIV-1 integrase inhibitory activity;

(5) Synthesis of 3-(carboxymethyl)-5-methoxy-1H-indole-2-carboxylic acid (compound f with HIV-1 integrase inhibitory activity): to a mixture containing acetone: water as 2: 1. Add 78.60 mg of sodium hydroxide to the reactor of 100.00 mg of substance e and stir the reaction at room temperature. When the reaction of the raw materials is completed, the reaction can be processed, that is, pour the reaction solution into a beaker, adjust the pH to weakly alkaline, and extract with ethyl acetate. 4 times, concentrated and separated by silica gel column chromatography to obtain compound f with HIV-1 integrase inhibitory activity.

In the reaction process of the steps (1)-(4), thin layer chromatography (Thin Layer Chromatography, TLC) is performed to monitor the progress of the reaction, and if the reaction is not complete, the reaction is continued until the reaction is complete.

The described experimental post-treatment is to adjust the pH to weak alkaline, extract three times with ethyl acetate (EthylAcetate, EA), concentrate the extract, pack into a column, and separate and purify the target product by thin layer chromatography.

The described pH adjustment is weakly alkaline, and is adjusted with anhydrous sodium carbonate.

Compound f with HIV-1 integrase inhibitory activity

Example 5

Take compound f with HIV-1 integrase inhibitory activity obtained in Example 4, in a 100 mL reactor, mix 50.00 mg of compound f with HIV-1 integrase inhibitory activity and 15 mL of distilled water, stir magnetically, and heat in an oil bath to 40 ℃, then add 30% sodium hydroxide solution to control the pH value between 11-11.5, heat to 85-95 ℃ to dissolve it, add a small amount of activated carbon, decolorize for 15 minutes, suction filtration, the filtrate is cooled to 25 ℃ and filtered , let the suction filtrate cool to room temperature for natural crystallization, wash the crystals with a small amount of ice water, and dry at 80°C to obtain compound g with HIV-1 integrase inhibitory activity.

Compound g with HIV-1 integrase inhibitory activity

Example 6

The raw material a in Example 4 was replaced by 3-hydroxy-4-methoxyaniline with 3-hydroxy-4-methoxyaniline, and the remaining steps were the same, and finally compound h with HIV-1 integrase inhibitory activity was obtained.

Compound h with HIV-1 integrase inhibitory activity

Example 7

The raw material 2 acyl glutaric acid diethyl ester in Example 4 was replaced with 2-acetyl-5-methoxy-4-oxoglutaric acid, and the remaining steps were the same, and finally obtained with HIV-1 integrase inhibitory activity of compound i.

Compound i with HIV-1 integrase inhibitory activity

Example 8

The raw material 2 acyl glutaric acid diethyl ester in the embodiment 4 is replaced with 2-acetyl-5-ethoxy-4-oxoglutaric acid, and all other steps are the same, and finally obtain HIV-1 integrase inhibitory activity the compound j.

Compound j with HIV-1 integrase inhibitory activity

Example 9

The raw material a in Example 4 was replaced with 3-fluoro-4-methoxyaniline, and the remaining steps were the same, and finally compound k with HIV-1 integrase inhibitory activity was obtained.

Compound k with HIV-1 integrase inhibitory activity

Example 10

The raw material a p-toluidine in Example 4 was replaced with 3-chloro-4-methoxyaniline, and the remaining steps were the same, and finally compound 1 with HIV-1 integrase inhibitory activity was obtained.

Compound 1 with HIV-1 integrase inhibitory activity

The compounds with HIV-1 integrase inhibitory activity obtained in Examples 1-10 were determined as follows:

(1) Integrase strand transfer activity assay

This experiment was carried out in a black polystyrene 96-well microplate in the dark, and the reaction system was 50 μL. Wash the plate once with 1× reaction buffer (25mM PIPES, 10mM β-mercaptoethanol, 0.1 g/LBSA, 10mM MnCl ₂ , 5% glycerol, pH 7.0), then add 25 μL of 2× reaction buffer without MnCl ₂ to each well solution, 550 nM purified integrase recombinant protein and 5 μL of the test compound dissolved in DMSO, mixed well and incubated at 37°C for 20 min. The positive control group was DTG. Then add MnCl ₂ with a final concentration of 10 mM, 30 nM donor DNA and 300 nM target DNA, and add sterile double-distilled water to make up the volume to 50 μL, and incubate at 37° C. for 1 h after mixing. Then add 1.5 μL of 10 mg/mL streptavidin magnetic beads and 51.5 μL of magnetic bead binding buffer (10 mM Tris-HCl, 2 M NaCl, 20 mM EDTA, 0.1% Tween-20, pH 7.6), thoroughly shake and mix Incubate at 20°C for 15 min, shake and mix every 5 min, place the microplate in a magnetic bead collector for 90 s, discard the supernatant, and wash the magnetic beads three times with PBS containing 0.05% Tween-20. Finally, the relative fluorescence value (RFU) of each well was detected by a fluorescence analyzer, the excitation wavelength was 485 nm, and the emission wavelength was 528 nm, and the inhibition rate of the sample to be tested was calculated.

(2) Cytotoxicity test of compounds on C8166 and PBMC

The compounds to be tested were diluted 5-fold in RPMI-1640 complete medium (containing 10% FBS) on a 96-well microplate, with a total of 6 dilutions, each dilution was set to 3 wells, and each well was 100 μL. At the same time, control wells without drug were set up. 4×10 ⁵ /mL of C8166 cells or 100 μL of 5×10 ⁶ /mL of PBMC were added to each well. The cells were incubated at 37°C in 5% _CO for 3 days, and the cytotoxicity was detected by MTT colorimetry. The OD value was measured by a microplate reader, and the measurement wavelength was 595 nm, and the CC ₅₀ value was calculated.

(3) Inhibitory experiments of compounds on HIV-1 _IIIB cytopathic (CPE)

The compounds to be tested were diluted 5-fold with RPMI-1640 complete medium on a 96-well microplate (the initial concentration was 20 μM), and three replicate wells were set for each dilution, with 100 μL in each well, and no Control wells for the drug. Add 50 μL of 8×10 ⁵ /mL C8166 cells to each well, and then add 50 μL of HIV-1 _IIIB diluted supernatant, 1300 TCID 50/well. The cells were incubated for 3 days at 37°C, 5% CO ₂ , and the formation of syncytia was counted under an inverted microscope (100×). _EC50 is the concentration of compound that inhibits syncytia formation by 50%.

The activities of the compounds with HIV-1 integrase inhibitory activity obtained in Examples 1-10 are as follows:

The above activity test results show that the compound with HIV-1 integrase inhibitory activity of the present invention has significant antiviral activity and low cytotoxicity, which provides a new option for the development of AIDS drugs.

Finally, it should be pointed out that the above embodiments are only representative examples of the present invention. Obviously, the technical solutions of the present invention are not limited to the above-mentioned embodiments, and many modifications are possible. All deformations that those of ordinary skill in the art can directly derive or associate from the disclosure of the present invention shall be considered as the protection scope of the present invention.

Claims (6)

1. The application of a compound with HIV-1 integrase inhibitory activity in preparing anti-HIV drugs is characterized in that the compound is represented by the following general formula (I):

in the general formula (I), R1 is OH, R2 is OCH₂CH₃And R3 is H.

2. The process for the preparation of a compound having HIV-1 integrase inhibitory activity according to claim 1 comprising the steps of:

(1) synthesizing p-methoxy diazobenzene hydrochloride: dissolving 1 time equivalent p-methylaniline with concentrated hydrochloric acid completely, stirring for 2h at room temperature, transferring into a reaction system at 0-10 ℃, slowly adding 2 times equivalent sodium nitrite when the temperature is constant, stirring for reaction, and reacting for 2-5h at the temperature to obtain p-methoxy diazobenzene hydrochloride;

(2) synthesis of diethyl (Z) -2-acetyl-2- ((4-methoxyphenyl) diazenyl) glutarate: dissolving 1.2 times of raw material diethyl 2-acetylglutamate with absolute ethyl alcohol completely, adding excessive concentrated H₂SO₄Fully and uniformly mixing the raw materials, slowly and constantly dropping a solution of 1-time equivalent of p-methoxy diazobenzene hydrochloride, reacting at room temperature for 2-3h after the dropping is finished, and carrying out experimental post-treatment after the reaction is finished, namely separating and purifying to obtain (Z) -2-acetyl-2- ((4-methoxyphenyl) diazenyl) diethyl glutarate;

(3) synthesis of (Z) -2- (2- (4-methoxyphenyl) hydrazono) glutaric acid diethyl ester: completely dissolving the separated diethyl (Z) -2-acetyl-2- ((4-methoxyphenyl) diazenyl) glutarate by using absolute ethyl alcohol, adding excessive concentrated HCl, stirring at room temperature for 2-5h, carrying out experimental post-treatment after the reaction is finished, and then separating and purifying to obtain diethyl (Z) -2- (2- (4-methoxyphenyl) hydrazono) glutarate;

(4) synthesis of 3- (2-ethoxy-2-oxoethyl) -5-methoxy-1H-indole-2-carboxylic acid ethyl ester: and (Z) -2- (2- (4-methoxyphenyl) hydrazono) diethyl glutarate is completely dissolved by absolute ethyl alcohol, dried HCl gas is introduced, the mixture is stirred for 2 to 7 hours at room temperature, after the reaction is finished, experimental post-treatment is carried out, and then the ethyl 3- (2-ethoxy-2-oxoethyl) -5-methoxy-1H-indole-2-carboxylate is separated and purified.

3. The method for preparing a compound having HIV-1 integrase inhibitory activity according to claim 2 wherein during the reaction in steps (1) to (4), thin layer chromatography is performed to monitor the progress of the reaction, and if the reaction is not complete, the reaction is continued until the reaction is complete.

4. The method for preparing a compound having an inhibitory activity against HIV-1 integrase according to claim 2, wherein the post-experimental treatment comprises adjusting the pH of the reaction solution, extracting with ethyl acetate, concentrating, and separating.

5. The method for preparing a compound having HIV-1 integrase inhibitory activity according to claim 4, wherein the steps of adjusting pH of the reaction solution, extracting with EA, concentrating, separating, specifically adjusting pH to weak alkaline, extracting with EA, concentrating the extract, packing with a column, and separating and purifying the target product by silica gel column chromatography.

6. The process for preparing a compound having HIV-1 integrase inhibitory activity according to claim 5, wherein the adjustment of the pH to a slightly alkaline pH is carried out using anhydrous sodium carbonate.

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