Pyrrolo [2, 3-d] pyrimidine derivative of aryl fragment and piperidine fragment as well as preparation method and application of pyrrolo [2, 3-d] pyrimidine derivative

Abstract

The invention belongs to the technical field of medicines, and particularly relates to pyrrolo [2, 3-d] pyrimidine compounds of aryl fragments and piperidine fragments as well as a preparation method and application of the pyrrolo [2, 3-d] pyrimidine compounds. The pyrrolo [2, 3-d] pyrimidine compound containing the aryl fragment and the piperidine fragment is an HIV-1 reverse transcriptase inhibitor, and the pyrrolo [2, 3-d] pyrimidine compound further comprises pharmaceutical salt, hydrate and solvate of the pyrrolo [2, 3-d] pyrimidine compound, polycrystal or eutectic of the pyrrolo [2, 3-d] pyrimidine compound, and precursors and derivatives with the same biological functions of the pyrrolo [2, 3-d] pyrimidine compound. The invention also relates to a preparation method of the compound and application of a composition containing one or more of the compounds in related medicines for treating AIDS and the like. The result of an in-vitro cellular level anti-HIV-1 activity experiment shows that the micromolecule has relatively strong anti-HIV-1 biological activity, can remarkably inhibit virus replication in MT-4 cells infected by HIV-1 viruses, and has relatively low cytotoxicity.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a pyrrolo[2,3-d]pyrimidine derivative containing aryl and piperidine fragments, its preparation method, and its application. Background Technology

[0002] Human immunodeficiency virus (HIV) is the culprit behind AIDS (acquired immunodeficiency syndrome). HIV attacks the body's T lymphocytes, disrupting cellular and humoral immune processes, thus causing the immune system to lose its function. According to data from UNAIDS in 2024, there were 40.8 million people living with HIV globally, with 1.3 million new infections in 2024, and 630,000 deaths from AIDS in 2024.

[0003] The HIV life cycle consists of the following five steps: (1) adsorption and gradual fusion with host T lymphocytes, releasing genomic RNA into the host cell; (2) reverse transcription of RNA into DNA under the action of reverse transcriptase; (3) integration of viral DNA into the host genome; (4) transcription and translation using enzymes and substances within the host cell to synthesize the genome and proteins required by the virus; (5) assembly within the host and release outside the host cell. These viruses continue to infect new host cells, thereby damaging the host's immune system. Several key enzymes are involved in the entire life cycle: fusion enzymes, reverse transcriptases, proteases, and integrases. Among them, reverse transcriptase (RT) plays a crucial role and is an important target for designing anti-HIV-1 drugs. Currently, there are 14 marketed reverse transcriptase inhibitors.

[0004] RT inhibitors can be divided into nucleoside reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs). Nucleoside reverse transcriptase inhibitors competitively bind to the RT active site, resulting in poor selectivity and high toxicity. Non-nucleoside reverse transcriptase inhibitors, on the other hand, bind non-competitively to an allosteric binding pocket approximately 10 Å away from the reverse transcriptase active site, known as the non-nucleoside reverse transcriptase inhibitor binding pocket (NNIBP). NNRTIs are characterized by high selectivity and high activity. Currently, the NNRTIs used clinically are mainly second-generation inhibitors: diarylpyrimidine compounds, such as rilpivirine (RPV) and etravirine (ETR).

[0005] However, poor water solubility (ETR, << 1 μg / mL; RPV, 20 ng / mL), low patient response rates (ETR, 36.5%; RPV, 27.3%), and the emergence of drug-resistant viral strains during long-term use significantly reduce the efficacy of the drug. Furthermore, toxic side effects occurring in clinical use (such as cardiotoxicity of ETR (hERG toxicity, IC50)) 50 The limited use of non-nucleoside reverse transcriptase inhibitors (NSTs) due to their low concentration (e.g., 0.5 μM) and their inhibitory effect on CYP enzymes (RPV) restricts their clinical application. Therefore, developing novel, highly effective, and low-toxicity non-nucleoside reverse transcriptase inhibitors with broad-spectrum anti-drug resistance has become a hot research topic for medicinal chemists.

[0006] This invention aims to optimize the structure of RPV and ETR by retaining the aromatic ring and pyrimidine core on the left side, introducing a pyrrole ring to occupy the entrance channel region, and introducing a piperidinyl group to replace the p-cyanophenyl group, thereby improving the safety and drug-likeness of the compound, enhancing the interaction between the compound and the amino acids inside the NNIBP pocket, and improving the bioactivity of the compound against drug-resistant viral strains. Summary of the Invention

[0007] The purpose of this invention is to propose a reverse transcriptase inhibitor with strong anti-HIV-1 bioactivity and low cytotoxicity, consisting of aryl and piperidine fragments of pyrrolo[2,3- d Pyrimidine derivatives, their preparation methods, and applications.

[0008] The reverse transcriptase inhibitor provided by this invention has strong anti-HIV-1 biological activity and low cytotoxicity. It is composed of aryl and piperidine fragments and is formulated with pyrrolo[2,3- d Pyrimidines are derivatives of the parent nucleus, specifically including pyrrolo[2,3-] ... d Pyrimidine derivatives, their medicinal salts, their hydrates and solvates, their polycrystalline and eutectic forms, their precursors and derivatives with similar biological functions; ; Wherein, R1 is selected from hydrogen, cyano, nitro, amino, hydroxyl, halogen, sulfonic acid, sulfonyl, carboxyl, ester, amide, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Alkyl mercapto, C 3~6 cycloalkyl, C 3~6 Cycloalkoxy, C 3~6 Cycloalkylamino, C 2~6 alkenyl, C 2~6 alkynyl group, optionally substituted C 1~6 Alkyl group, optionally substituted C 1~6 Alkoxy group, optionally substituted C 1~6 Alkyl mercapto group, optionally substituted C3~6 cycloalkyl, with optional substituted C 3~6 Cycloalkoxy, optionally substituted C 3~6 Cycloalkylamino, with the C-terminus optionally substituted 2~6 alkenyl group, with the C group optionally substituted. 2~6 alkynyl or C 1~6 Alkoxy; R2 is selected from hydrogen, cyano, nitro, amino, hydroxyl, halogen, sulfonic acid, sulfonyl, carboxyl, ester, amide, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Alkyl mercapto, C 3~6 cycloalkyl, C 3~6 Cycloalkoxy, C 3~6 Cycloalkylamino, C 2~6 alkenyl, C 2~6 alkynyl group, optionally substituted C 1~6 Alkyl group, optionally substituted C 1~6 Alkoxy group, optionally substituted C 1~6 Alkyl mercapto group, optionally substituted C 3~6 cycloalkyl, with optional substituted C 3~6 Cycloalkoxy, optionally substituted C 3~6 Cycloalkylamino, with the C-terminus optionally substituted 2~6 alkenyl group, with the C group optionally substituted. 2~6 alkynyl or C 1~6 Alkoxy; R2 can also be a mono- or poly-substituted aromatic ring, with the substituents on the aromatic ring selected from hydrogen, cyano, nitro, amino, hydroxyl, halogen, sulfonic acid, sulfonyl, carboxyl, ester, amide, C. 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Alkyl mercapto, C 3~6 cycloalkyl, C 3~6 Cycloalkoxy, C 3~6 Cycloalkylamino, C 2~6 alkenyl, C 2~6 alkynyl group, optionally substituted C 1~6 Alkyl group, optionally substituted C 1~6 Alkoxy group, optionally substituted C 1~6 Alkyl mercapto group, optionally substituted C 3~6 cycloalkyl, with optional substituted C 3~6 Cycloalkoxy, optionally substituted C 3~6 Cycloalkylamino, with the C-terminus optionally substituted 2~6 alkenyl group, with the C group optionally substituted. 2~6 alkynyl or C 1~6 Alkoxy; Ar is a mono- or poly-substituted aromatic ring, and the substituents on the aromatic ring are selected from hydrogen, cyano, nitro, amino, hydroxyl, halogen, sulfonic acid group, sulfonyl group, various substituted sulfonyl groups, sulfonamide group, various substituted sulfonamide groups, carboxyl group, ester group, various substituted ester group, amide group, various substituted amide group, borate group, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Alkyl mercapto, C 3~6 cycloalkyl, C 3~6 Cycloalkoxy, C 3~6 Cycloalkylamino, C 2~6 alkenyl, C 2~6 alkynyl group, optionally substituted C 1~6 Alkyl group, optionally substituted C 1~6 Alkoxy group, optionally substituted C 1~6 Alkyl mercapto group, optionally substituted C 3~6 cycloalkyl, with optional substituted C 3~6 Cycloalkoxy, optionally substituted C 3~6 Cycloalkylamino, with the C-terminus optionally substituted 2~6 alkenyl group, with the C group optionally substituted. 2~6 alkynyl or C 1~6 Alkoxy; X is one of the following linking groups: -CH2-, -NH-, -O-, -S-, -S(O)-, and -S(O)2-.

[0009] In this invention, the aryl fragment and the piperidine fragment are pyrrolo[2,3- d Pharmaceutical salts of pyrimidine derivatives, including hydrochloride, hydrobromide, sulfate, phosphate, acetate, methanesulfonate, p-toluenesulfonate, tartrate, citrate, fumarate, or malate.

[0010] This invention also proposes pyrrolo[2,3-] ... d The preparation method of pyrimidine derivatives, with the following general reaction formula: ; The specific preparation steps are as follows: Using the compound shown in structural formula II as a raw material, compound I was prepared by reacting it with the corresponding halomethylaryl group (compound III) in a solvent under alkaline conditions. The solvents mentioned are water, dichloromethane, chloroform, ethyl acetate, methanol, ethanol, n-propanol, isopropanol, n-butanol, tert-butanol, acetonitrile, tetrahydrofuran, nitromethane, acetic anhydride, naphthane, toluene, etc. N , N -Dimethylformamide, N , N -Dimethylacetamide,N One or more of methylpyrrolidone, preferably N , N -Dimethylformamide; The alkali is one of the following inorganic bases: sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide, sodium hydroxide, or sodium hydroxide; or one of the following organic bases: N , N - One or more of dimethylaminopyridine, triethylamine, diisopropylethylamine, tributylamine, potassium tert-butoxide, sodium tert-butoxide, sodium methoxide, and sodium ethoxide, preferably potassium carbonate; The molar ratio of compound II, compound III to the base is 1:1:1.05 to 1:2:5, preferably 1:1.05:1.1 to 1:1.5:2; The reaction temperature is room temperature to 100 ℃, preferably 20 to 40 ℃; The reaction time is 0.5 to 24 hours, preferably 2 to 6 hours.

[0011] The present invention also provides a pharmaceutical composition comprising an effective dose of the above-mentioned pyrrolo[2,3-] ... d Pyrimidine derivatives, or their pharmaceutical salts, or their hydrates and solvates, or their polycrystalline and eutectic forms, and pharmaceutical carriers.

[0012] This invention also provides pyrrolo[2,3-] ... d The use of pyrimidine derivatives or compositions in the preparation of drugs for the prevention and treatment of AIDS.

[0013] This invention is based on the binding mode of diarylpyrimidine derivatives to reverse transcriptase, combined with computer-aided drug design, by introducing pyrrolo[2,3-] ... d The pyrimidine core and benzylpiperidine structure, with the newly introduced pyrrolo[2,3- d The pyrimidine core can form hydrogen bonds with surrounding amino acids such as K101 via water molecules, and the benzylpiperidine fragment can form a hydrogen bond network with K103, K104, and P236, significantly enhancing the binding force with reverse transcriptase and further improving the antiviral activity of the target compounds. Bioactivity tests showed that all compounds exhibited strong anti-HIV-1 activity and low cytotoxicity, with a high selectivity index.

[0014] Pyrrolo[2,3-]py ... d Pyrimidine nuclei. Detailed Implementation

[0015] The following examples can provide a better understanding of the invention, but they do not limit the scope of the invention.

[0016] Example 1: Synthesis of final product Ia At 20–40 °C, compound II and the base were added to the solvent, and compound III was added while stirring. After the addition was complete, stirring was continued for 2–6 h. The reaction was monitored by TLC until compound II was completely consumed. The reaction solution was poured into ethyl acetate and washed successively with water and saturated brine. The organic phase was dried overnight with anhydrous sodium sulfate. The solution was filtered, concentrated, and separated by column chromatography to obtain the desired solid.

[0017] The target compound was prepared by the above method using different compounds II, and some results are as follows: At room temperature, 3',5'-dimethyl-4'-((2-(piperidin-4-ylamino)-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)oxy)-[1,1'-biphenyl]-4-carbonitrile (0.5 mmol), potassium carbonate (0.75 mmol) dissolved in 5 mL N , N -Dimethylformamide. Subsequently, 2-(bromomethyl)naphthalene (0.6 mmol) was added to the reactor under stirring. After the addition was complete, the reaction was stirred at room temperature for 3 h. The reaction was monitored for completion. The reaction was stopped, and the mixture was extracted with ethyl acetate and water. The organic layer was collected, dried over anhydrous sodium sulfate, evaporated to dryness, and separated by column chromatography (eluting with dichloromethane and methanol) to obtain a white solid, designated as Ia.

[0018] Ⅰa; White solid; yield 16%. 1 H NMR (400 MHz, DMSO- d 6) d 11.32 (s, 1H), 7.91 (q, J = 8.7 Hz, 4H), 7.87 – 7.80 (m, 3H), 7.73 – 7.61 (m, 1H), 7.54 (s, 2H), 7.51– 7.44 (m, 2H), 7.44 – 7.37 (m, 1H), 6.90 (d, J = 3.6 Hz, 1H), 6.38 (d, J=7.5 Hz, 1H), 6.16 (s, 1H), 3.56 – 3.48 (m, 2H), 2.74 (s, 2H), 2.13 (s, 6H), 1.89 (s, 1H), 1.73 (s, 2H), 1.40 (s, 2H). 13 C NMR (101 MHz, DMSO- d 6) d 161.64, 158.48, 156.56, 151.23, 144.73, 136.93, 135.53, 133.31, 133.28, 132.69, 131.96, 128.07, 127.95, 127.86, 127.70, 127.57, 127.38, 126.47, 126.04, 120.58, 119.42, 110.20, 98.41, 96.67, 62.86, 53.05, 31.99, 16.79.

[0019] Example 2, Synthesis of final product Ib Ⅰb; The procedure was the same as in Example 1. White solid; yield 47%. 1 H NMR (400 MHz, DMSO- d 6) d 11.32 (s, 1H), 8.38 (q, J = 4.5 Hz, 1H), 7.92 (q, J = 8.4 Hz, 4H), 7.76 (d, J = 7.8 Hz,2H), 7.55 (s, 2H), 7.29 (s, 2H), 6.95 – 6.84 (m, 1H), 6.37 (d, J = 7.5 Hz,1H), 6.16 (s, 1H), 2.78 (d, J = 4.5 Hz, 3H), 2.69 (s, 2H), 2.13 (s, 6H), 1.86 (s, 1H), 1.79 – 1.67 (m, 2H), 1.40 (s, 2H). 13 C NMR (101 MHz, DMSO- d 6) d166.99, 161.64, 158.47, 151.23, 144.71, 142.38, 135.52, 133.63, 133.28, 131.96, 128.89, 127.85, 127.58, 127.41, 120.57, 119.41, 110.21, 98.42, 96.67, 62.27, 52.98, 31.96, 26.68, 16.80.

[0020] Example 3, Synthesis of final product Ic Ⅰc; The procedure was the same as in Example 1. White solid; yield 57%. 1 H NMR (400 MHz, DMSO- d 6) d 11.33 (s,1H), 8.11 – 7.74 (m, 7H), 7.56 (s, 3H), 7.40 (s, 2H), 6.92 (t, J = 2.9 Hz,1H), 6.54 (s, 1H), 6.15 (s, 1H), 3.81 – 3.41 (m, 4H), 3.07 (q, J = 7.3 Hz,1H), 2.14 (s, 6H), 1.96 – 1.74 (m, 2H), 1.55 (s, 2H), 1.20 (t, J = 7.3 Hz, 1H). 13 C NMR (101 MHz, DMSO- d 6) d 161.68, 158.35, 151.18, 144.67, 135.55, 133.31, 131.95, 128.05, 127.86, 127.63, 119.40, 110.24, 98.45, 45.95, 16.79, 8.98.

[0021] Example 4, Synthesis of final product Id Ⅰd; The procedure was the same as in Example 1. White solid; yield 44%. 1 H NMR (400 MHz, DMSO- d 6) d11.33 (s,1H), 7.98 – 7.86 (m, 4H), 7.71 (d, J = 7.9 Hz, 2H), 7.55 (s, 2H), 7.51 – 7.35(m, 3H), 6.90 (dd, J = 3.6, 1.8 Hz, 1H), 6.39 (d, J = 7.5 Hz, 1H), 6.16 (s,1H), 3.45 (s, 2H), 2.70 (s, 2H), 2.40 (s, 3H), 2.13 (s, 6H), 1.89 (s, 1H),1.74 (s, 2H), 1.48 – 1.32 (m, 2H). 13 C NMR (101 MHz, DMSO- d 6) d 161.65, 158.47, 156.55, 151.23, 144.71, 144.17, 138.17, 135.51, 133.29, 131.96, 129.60, 127.84, 127.59, 127.08, 120.59, 119.40, 110.21, 98.42, 96.68, 62.01, 53.01, 31.95, 29.15, 16.80.

[0022] Example 5, Synthesis of final product Ie Ⅰe; The procedure was the same as in Example 1. The product was a white solid; yield 30%. 1 H NMR (400 MHz, DMSO- d 6) d 11.32 (s, 1H), 8.66 (t, J = 5.7 Hz, 1H), 7.98 – 7.85 (m, 5H), 7.77 (d, J = 7.9 Hz, 1H),7.53 (s, 2H), 7.32 (d, J = 24.3 Hz, 2H), 6.89 (d, J = 3.6 Hz, 1H), 6.36 (dd, J = 7.7, 3.7 Hz, 1H), 6.20 – 6.08 (m, 1H), 4.39 (t, J= 5.6 Hz, 1H), 3.64 (q, J = 5.6 Hz, 1H), 2.68 (s, 2H), 2.12 (s, 6H), 1.85 (s, 1H), 1.72 (s, 2H), 1.49– 1.28 (m, 2H). 13 C NMR (101 MHz, DMSO- d 6) d 166.97, 166.15, 161.64, 158.46, 156.54, 151.22, 145.04, 144.69, 142.65, 135.50, 133.27, 131.95, 129.69, 129.18, 128.91, 127.83, 127.56, 120.56, 119.39, 110.20, 98.41, 96.67, 63.78, 62.24, 52.96, 31.95, 16.79.

[0023] Example 6, Synthesis of final product If Ⅰf; The procedure was the same as in Example 1. White solid; yield 46%. 1 H NMR (400 MHz, DMSO- d 6) d 11.30 (s, 1H), 7.93 (d, J = 8.5 Hz, 2H), 7.92 – 7.85 (m, 4H), 7.54 (s, 2H), 7.37 (s,2H), 6.89 (dd, J = 3.6, 2.2 Hz, 1H), 6.37 (d, J = 7.4 Hz, 1H), 6.15 (s, 1H), 3.84 (s, 3H), 2.75 – 2.61 (m, 2H), 2.13 (s, 6H), 1.89 (s, 1H), 1.72 (s, 2H), 1.39 (s, 2H). 13 C NMR (101 MHz, DMSO- d 6) d166.63, 161.64, 158.46, 151.24, 145.04, 144.69, 135.50, 133.28, 131.96, 129.54, 129.28, 128.65, 127.83, 127.56, 119.41, 110.18, 96.68, 62.21, 52.98, 52.51, 31.94, 16.78.

[0024] Example 7, Synthesis of final product Ig Ⅰg; The procedure was the same as in Example 1. White solid; yield 36%. 1 H NMR (400 MHz, DMSO- d 6) d 11.32 (s,1H), 7.98 – 7.86 (m, 4H), 7.55 (s, 2H), 7.28 (s, 4H), 6.90 (dd, J = 3.6, 2.0Hz, 1H), 6.38 (d, J = 7.5 Hz, 1H), 6.16 (s, 1H), 4.81 (d, J = 3.7 Hz, 1H),3.73 (s, 2H), 3.16 (s, 3H), 2.71 (s, 2H), 2.13 (s, 6H), 1.83 (s, 2H), 1.73(s, 3H), 1.39 (d, J = 17.8 Hz, 4H). 13 C NMR (101 MHz, DMSO- d 6) d 169.38, 161.65, 158.47, 156.55, 144.71, 140.53, 135.50, 135.31, 133.29, 131.96, 128.98, 127.84, 127.58, 127.03, 119.41, 110.20, 96.67, 65.98, 62.33, 60.23, 53.00, 31.98, 21.24, 16.80, 14.55.

[0025] Example 8, Synthesis of final product Ih Ⅰh; The procedure was the same as in Example 1. White solid; yield 76%.1 H NMR (400 MHz, DMSO- d 6) d 11.31 (s, 1H), 7.93 (q, J = 8.5 Hz, 4H), 7.76 (d, J = 8.2 Hz, 2H), 7.55 (s, 2H), 7.41(s, 2H), 7.30 (s, 2H), 6.90 (dd, J = 3.6, 2.2 Hz, 1H), 6.39 (d, J = 7.5 Hz,1H), 6.15 (s, 1H), 3.43 (s, 2H), 3.18 (d, J = 4.1 Hz, 1H), 2.69 (s, 2H), 2.14(s, 6H), 1.87 (s, 1H), 1.74 (s, 2H), 1.40 (s, 2H). 13 C NMR (101 MHz, DMSO- d 6) d 161.65, 158.46, 144.72, 143.11, 135.52, 133.30, 131.96, 129.41, 127.85, 127.60, 126.03, 119.41, 110.22, 96.67, 62.03, 52.96, 31.93, 16.80.

[0026] Example 9, Synthesis of final product Ii Ⅰi; The procedure was the same as in Example 1. White solid; yield 67%. 1 H NMR (400 MHz, DMSO- d 6) d 11.31 (s, 1H), 7.92 (q, J = 8.6, 8.1 Hz, 5H), 7.75 (d, J = 7.2 Hz, 2H), 7.55 (s, 2H), 7.35 (d, J = 10.8 Hz, 3H), 6.93 – 6.84 (m, 1H), 6.38 (d, J= 7.5 Hz, 1H), 6.15 (s, 1H), 3.50 – 3.42 (m, 2H), 2.72 (s, 2H), 2.13 (s, 6H), 1.93 (d, J =34.2 Hz, 1H), 1.74 (s, 2H), 1.40 (s, 2H). 13 C NMR (101 MHz, DMSO- d 6) d 168.45, 161.65, 158.47, 151.21, 144.71, 135.53, 134.65, 133.29, 132.11, 131.96, 128.44, 127.85, 127.60, 126.43, 119.41, 110.21, 98.44, 96.67, 52.89, 31.89, 16.80.

[0027] Example 10, Synthesis of final product Ij Ⅰj; The procedure was the same as in Example 1. White solid; yield 58%. 1 H NMR (400 MHz, DMSO- d 6) d 11.32 (s, 1H), 7.92 (q, J = 8.4 Hz, 4H), 7.55 (s, 2H), 7.10 (s, 2H), 6.89 (d, J = 3.7Hz, 1H), 6.83 (d, J = 8.1 Hz, 2H), 6.35 (d, J = 7.4 Hz, 1H), 6.16 (s, 1H), 3.72 (s, 3H), 3.27 (s, 2H), 2.68 (s, 2H), 2.13 (s, 6H), 1.71 (s, 2H), 1.35(s, 2H). 13 C NMR (101 MHz, DMSO- d 6) d161.64, 158.61, 158.47, 151.24, 144.70, 135.49, 133.29, 131.95, 130.92, 130.36, 127.84, 127.56, 120.57, 119.41, 113.90, 110.19, 98.41, 96.66, 62.12, 55.43, 52.79, 31.97, 16.79.

[0028] Example 11: Anti-HIV bioactivity test The in vitro cellular anti-HIV activity was determined by the Rega Institute of Pharmaceutical Research at KU Leuven, Belgium, primarily including inhibitory activity and cytotoxicity against HIV-infected MT-4 cells. The method was as follows: the compound was administered to HIV-infected MT-4 cells at different time points after HIV infection. The protective effect of the drug against HIV-induced cytopathic effects was determined using the MTT assay. The half-maximal effective concentration (EC50) required to protect 50% of the cells from HIV-induced cytopathic effects was calculated. 50 The toxicity assay and anti-HIV activity assay were performed in parallel, also in MT-4 cell culture, using the MTT assay to determine the concentration (CC) that caused cytopathic effects in 50% of uninfected cells. 50 ), and calculate the selectivity index SI = CC 50 / EC 50 .

[0029] Materials and Methods: The anti-HIV activity of each compound was monitored by its efficiency in inhibiting HIV-induced cytopathic effects in cells. MT-4 cells were used for cell culture. The viral strains used were HIV-1 strain IIIB and HIV-2 strain ROD.

[0030] The specific procedure is as follows: Dissolve the compound in DMSO or water, then dilute it in a phosphate buffered saline solution, adding 3×10... 5 MT-4 cells were treated with 100 μL of solutions containing different concentrations of various compounds at 37°C. o Pre-culture at 37°C for 1 hour, then add 100 μL of appropriate virus dilution to the compound, and incubate the cells at 37°C. o Incubate at C for 1 hour. After washing three times, resuspend the cells in culture medium containing or without the compound. Then, incubate the cells at 37°C under a 5% CO2 atmosphere. oThe culture medium was incubated at C for 7 days, and replenished with culture medium containing or without the compound on the third day post-infection. Each culture medium condition was repeated twice. The cytopathic effect of the virus was monitored daily using a reverse optical microscope. Typically, the virus dilution used in this experiment often caused cytopathic effects by the fifth day post-infection. The drug inhibitory concentration was defined as the concentration at which the drug produced 50% inhibition of the viral cytopathic effect without direct cytotoxicity to the cells (CC). 50 It is important to emphasize that when a compound has poor water solubility and requires DMSO to dissolve, the DMSO concentration relative to water is generally less than 10% (the final concentration of DMSO in MT-4 cell culture medium is less than 2%). Because DMSO can affect the antiviral activity of the tested compound, antiviral activity comparison experiments with blanks containing the same concentration of DMSO should also be performed in parallel. Furthermore, the final concentration of DMSO (1 / 1000) is far lower than the concentration required for HIV-1 replication in T cells.

[0031] This invention uses marketed drugs nevirapine (NVP), efavirenz (EFV), and etravirine (ETR) as reference standards. The results of the inhibitory activity of some target compounds against HIV are shown in Table 1 (Anti-HIV activity and cytotoxicity of compounds 1-19 in MT-4 cells).

[0032] .

[0033] Table 1 [a] .

[0034] [a] All data represent mean values ​​of at least three separate experiments. [b] EC 50 : effective concentration required to protect 50% of cells against viral cytopathicity in MT-4 cells. [c] CC 50: cytotoxicconcentration of the compound that reduces the normal uninfected MT-4 cellviability by 50%. [d] SI: selectivity index, ratio CC 50 / EC 50 (WT).

[0035] Experimental results show that the compounds contained in the general chemical formula generally have strong anti-HIV-1 virus activity, low cytotoxicity, and high selectivity index.

[0036] The present invention is not limited to the above examples.

Claims

1. A pyrrolo[2,3-d]pyrimidine derivative comprising an aryl fragment and a piperidine fragment, characterized in that, Its structural formula is as follows: ； Wherein, R1 is selected from hydrogen, cyano, nitro, amino, hydroxyl, halogen, sulfonic acid, sulfonyl, carboxyl, ester, amide, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Alkyl mercapto, C 3~6 cycloalkyl, C 3~6 Cycloalkoxy, C 3~6 Cycloalkylamino, C 2~6 alkenyl, C 2~6 alkynyl group, optionally substituted C 1~6 Alkyl group, optionally substituted C 1~6 Alkoxy group, optionally substituted C 1~6 Alkyl mercapto group, optionally substituted C 3~6 cycloalkyl, with optional substituted C 3~6 Cycloalkoxy, optionally substituted C 3~6 Cycloalkylamino, with the C-terminus optionally substituted 2~6 alkenyl group, with the C group optionally substituted. 2~6 alkynyl or C 1~6 Alkoxy; R2 is selected from hydrogen, cyano, nitro, amino, hydroxyl, halogen, sulfonic acid, sulfonyl, carboxyl, ester, amide, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Alkyl mercapto, C 3~6 cycloalkyl, C 3~6 Cycloalkoxy, C 3~6 Cycloalkylamino, C 2~6 alkenyl, C 2~6 alkynyl group, optionally substituted C 1~6 Alkyl group, optionally substituted C 1~6 Alkoxy group, optionally substituted C 1~6 Alkyl mercapto group, optionally substituted C 3~6 cycloalkyl, with optional substituted C 3~6 Cycloalkoxy, optionally substituted C 3~6 Cycloalkylamino, with the C-terminus optionally substituted 2~6 alkenyl group, with the C group optionally substituted. 2~6 alkynyl or C 1~6 Alkoxy; Alternatively, R2 is a mono- or poly-substituted aromatic ring, with the substituents on the aromatic ring selected from hydrogen, cyano, nitro, amino, hydroxyl, halogen, sulfonic acid, sulfonyl, carboxyl, ester, amide, C. 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Alkyl mercapto, C 3~6 cycloalkyl, C 3~6 Cycloalkoxy, C 3~6 Cycloalkylamino, C 2~6 alkenyl, C 2~6 alkynyl group, optionally substituted C 1~6 Alkyl group, optionally substituted C 1~6 Alkoxy group, optionally substituted C 1~6 Alkyl mercapto group, optionally substituted C 3~6 cycloalkyl, with optional substituted C 3~6 Cycloalkoxy, optionally substituted C 3~6 Cycloalkylamino, with the C-terminus optionally substituted 2~6 alkenyl group, with the C group optionally substituted. 2~6 alkynyl or C 1~6 Alkoxy; Ar is a mono- or poly-substituted aromatic ring, and the substituents on the aromatic ring are selected from hydrogen, cyano, nitro, amino, hydroxyl, halogen, sulfonic acid group, sulfonyl group, various substituted sulfonyl groups, sulfonamide group, various substituted sulfonamide groups, carboxyl group, ester group, various substituted ester group, amide group, various substituted amide group, borate group, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Alkyl mercapto, C 3~6 cycloalkyl, C 3~6 Cycloalkoxy, C 3~6 Cycloalkylamino, C 2~6 alkenyl, C 2~6 alkynyl group, optionally substituted C 1~6 Alkyl group, optionally substituted C 1~6 Alkoxy group, optionally substituted C 1~6 Alkyl mercapto group, optionally substituted C 3~6 cycloalkyl, with optional substituted C 3~6 Cycloalkoxy, optionally substituted C 3~6 Cycloalkylamino, with the C-terminus optionally substituted 2~6 alkenyl group, with the C group optionally substituted. 2~6 alkynyl or C 1~6 Alkoxy; X is one of the following linking groups: -CH2-, -NH-, -O-, -S-, -S(O)-, and -S(O)2-.

2. A pharmaceutically acceptable salt of a pyrrolo[2,3-d]pyrimidine derivative of the aryl and piperidine fragments as described in claim 1, characterized in that, This includes hydrochloride, hydrobromide, sulfate, phosphate, acetate, methanesulfonate, p-toluenesulfonate, tartrate, citrate, fumarate, or malate, as well as pharmaceutically acceptable prodrugs and derivatives.

3. The method for preparing pyrrolo[2,3-d]pyrimidine derivatives of aryl and piperidine fragments as described in claim 1, characterized in that, The specific steps are as follows: Using the compound represented by structural formula II as a starting material, compound I was prepared by reacting it with the corresponding halomethylaryl group, i.e., compound III, in a solvent under alkaline conditions; the reaction formula is as follows: ； The solvent is one or more of the following: water, dichloromethane, chloroform, ethyl acetate, methanol, ethanol, n-propanol, isopropanol, n-butanol, tert-butanol, acetonitrile, tetrahydrofuran, nitromethane, acetic anhydride, naphthane, toluene, N,N-dimethylformamide, N,N-dimethylacetamide, and N-methylpyrrolidone. The base is one of the following inorganic bases: sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide, sodium hydrogen hydride, or one or more of the following organic bases: N,N-dimethylaminopyridine, triethylamine, diisopropylethylamine, tributylamine, potassium tert-butoxide, sodium tert-butoxide, sodium methoxide, sodium ethoxide. The molar ratio of compounds II and III to the base is 1:1:1.05 - 1:2:5; The reaction temperature is room temperature to 100℃; The reaction time is 0.5~24 h.

4. A pharmaceutical composition, characterized in that, The pharmaceutically acceptable derivatives of pyrrolo[2,3-d]pyrimidines containing an effective dose of the aryl and piperidine fragments as described in claim 1, or pharmaceutically acceptable salts thereof, or hydrates and solvates thereof, or polycrystalline and eutectic thereof, and pharmaceutically acceptable carriers thereof.

5. The pharmaceutical composition according to claim 4, characterized in that, The pharmaceutical salts include hydrochloride, hydrobromide, sulfate, phosphate, acetate, methanesulfonate, p-toluenesulfonate, tartrate, citrate, fumarate, or malate, as well as pharmaceutically acceptable prodrugs and derivatives.

6. The use of pyrrolo[2,3-d]pyrimidine derivatives of the aryl and piperidine fragments as described in claim 1 in the preparation of medicaments for the prevention and treatment of AIDS.

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