Dihydropteridine-6 (5H)-ketone skeleton compound as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a dihydropteridine-6 (5H)-ketone skeleton compound as well as a preparation method and application thereof. The dihydropteridine-6 (5H)-ketone skeleton compound disclosed by the invention is an HIV-1 (Human Immunodeficiency Virus-1) non-nucleoside reverse transcriptase inhibitor; the invention also comprises pharmaceutical salts, hydrates, solvates, polycrystals or co-crystals of the compounds, precursors and derivatives with the same biological functions of the compounds, a preparation method of the compounds and application of a composition containing one or more of the compounds in preparation of related drugs 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 method for preparing and using a diarylpyrimidine cyclic compound with a biaryl fragment. Background Technology

[0002] AIDS (Acquired Immunodeficiency Syndrome) is caused by the human immunodeficiency virus (HIV). HIV destroys the body's T lymphocytes, blocking cellular and humoral immune processes, leading to the paralysis of the immune system. According to a 2025 report by UNAIDS, as of the end of 2024, there were 40.8 million people living with HIV worldwide, with approximately 1.3 million new HIV infections that year, and about 630,000 deaths from AIDS-related illnesses—meaning someone dies from AIDS every minute on average.

[0003] The HIV life cycle can be summarized as follows: (1) it attaches to and gradually fuses with host T lymphocytes, releasing its genomic RNA into the host cell; (2) it forms DNA through reverse transcription and integrates it into the host's genome; (3) it uses enzymes and substances within the host cell to transcribe and translate, synthesizing the genome and proteins required by the virus; (4) it completes assembly within the host and is released 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 this life cycle: fusion enzymes, reverse transcriptases, proteases, and integrases. Among them, reverse transcriptase (RT) plays a crucial role and is also an important target for designing anti-HIV-1 drugs.

[0004] RT inhibitors can be divided into nucleoside reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs). NRTIs competitively act on the RT active site, while NNRTIs 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 affect viral DNA synthesis and replication by interfering with the RT catalytic active site, without interfering with normal human DNA synthesis. Therefore, they are highly selective and have low genotoxicity. Furthermore, they do not require conversion into active substances through host cell metabolism, resulting in faster onset of action and higher stability in vivo. Currently, the NNRTIs used clinically are mainly second-generation HIV inhibitors: etravirine (ETR) and rilpivirine (RPV), both of which have a diarylpyrimidine structure.

[0005] The diarylpyrimidine structure of ETR and RPV endows them with greater flexibility, enhancing the resistance barrier against mutant strains. However, this also determines their physicochemical properties, such as poor water solubility (S(ETR) << 1 μg / mL; S(RPV) = 20 ng / mL), resulting in low bioavailability. Furthermore, low patient response rates (ETR: 36.5%; RPV: 27.3%) and side effects from long-term use limit their clinical application. Moreover, long-term clinical use can lead to amino acid mutations in RT, causing the previously effective drug to lose its activity, resulting in drug-resistant HIV strains. Therefore, the development of novel, highly effective non-nucleoside reverse transcriptase inhibitors with broad-spectrum anti-drug resistance has become a hot research topic for medicinal chemists.

[0006] The present invention aims to optimize the structure of ETR and RPV by enhancing the nonpolar interaction between the compounds and the amino acids on the inner wall of NNIBP through biaryl fragments and pyrimidine cyclic rings, in order to improve the bioactivity of this series of compounds against drug-resistant viral strains. Summary of the Invention

[0007] The present invention aims to provide a dihydropteridine-6(5H)-one skeleton compound with potent inhibitory activity against HIV-1, which can significantly inhibit viral replication in HIV-1-infected MT-4 cells and has low cytotoxicity and significant safety, as well as its preparation method and uses.

[0008] The present invention provides a compound containing a dihydropteridine-6(5H)-one skeleton, the structural formula of which is as follows:

[0009] ;

[0010] Among them, R 1 Selected from cyano, cyanovinyl, various substituted or unsubstituted benzene rings, pyridine, pyrimidine, azines and their oxides, thiophene (furan) (sulfoxide and sulfone), (iso)thiazole and its oxides, pyrrole and its oxides, pyrazole (furan) and its oxides, imidazole and its oxides, (iso)oxazole and its oxides, pyrazolone, furan, cyclopentadiene, dihydrothiophene and its cyclic sulfides (sulfoxide and sulfone), dihydrofuran and its epoxides, and cycloalkanes;

[0011] R 2 Selected from H, CN, NO2, OH, CH2OH, CF3, F, Cl, Br, CH3, COOMe, COOH, SO2CH3, SONH2, CONH2 and a series of sulfonyl (derivatives) and amide (derivatives) with hydrophilic or lipophilic ends, with substituents being ortho, meta, para monosubstituted or polysubstituted;

[0012] X is a linker among -H-, -N-, -CH-, -S-, -S(O)-, and -S(O)2-.

[0013] Compound I of this class is an NNRTI, which not only has strong biological activity, but also low cytotoxicity and a high selectivity.

[0014] The compounds of the present invention also include pharmaceutically acceptable salts, stereochemical isomers, hydrates or solvates of derivatives.

[0015] In this invention, the pharmaceutically acceptable salt is a sodium salt, hydrochloride, hydrobromide, sulfate, formate, acetate, phosphate tartrate or citrate, succinate, maleate, trifluoromethanesulfonate, p-toluenesulfonate, methanesulfonate, fumarate or malate, etc.

[0016] This invention also provides a method for preparing the above-mentioned dihydropteridine-6(5H)-one skeleton compound, the reaction formula of which is:

[0017] ;

[0018] The specific preparation steps are as follows:

[0019] (i) In a solvent, using 2,6-dimethyl-4-bromoaniline (A) and 2,4-dichloro-5-nitro-pyrimidine (B) as raw materials, hydrogen is abstracted under the action of a base, and then reacted with ethyl bromoacetate to obtain compound C;

[0020] (ii) Subsequently, compound C reacts with N-Boc-4-aminopiperidine under appropriate solvent and alkaline conditions to give compound D;

[0021] (iii) After separation, compound D undergoes cyclization under reducing agent (iron powder) and acidic conditions (ammonium chloride aqueous solution) to give compound E;

[0022] (iv) Compound E was deprotected from its Boc protecting group in a mixed solvent of trifluoroacetic acid and dichloromethane to give compound F;

[0023] (v) Finally, compound F reacts with benzyl bromide or benzyl chloride with different functional groups in a solvent under the catalysis of a base to give compound II;

[0024] (vi) Compound II undergoes a metal coupling reaction to give the target product, dihydropteridine-6(5H)-one skeleton compound I.

[0025] Furthermore:

[0026] The solvent used in the entire reaction is one or more of N,N-dimethylformamide, N,N-dimethylacetamide, 1,4-dioxane, acetone, acetonitrile, toluene, dichloromethane, tetrahydrofuran, methanol, ethanol, isopropanol, n-butanol, and isobutanol, or the reaction can be carried out without solvent; among them, 1,4-dioxane is the best solvent for steps (i), (ii), and (vi); methanol is the best solvent for step (iii); dichloromethane is the best solvent for step (iv); and N,N-dimethylformamide is the best solvent for step (v).

[0027] The base used is one or more of sodium carbonate, sodium bicarbonate, potassium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide, sodium hydride, N,N-dimethylaminopyridine, triethylamine, diisopropylethylamine, tributylamine, potassium tert-butoxide, and sodium tert-butoxide; wherein sodium hydride is the best for step (i) reaction; and potassium carbonate is the best for steps (ii), (v), and (vi) reaction.

[0028] The acid used is one or more of hydrochloric acid, sulfuric acid, trifluoroacetic acid, formic acid, acetic acid, and ammonium chloride aqueous solution; among which, ammonium chloride aqueous solution is best for step (iii) and trifluoroacetic acid is best for step (iv).

[0029] The optimal reaction molar ratio of compounds A, B, ethyl bromoacetate, and base is 1:1:1.5:1~1:1.2:5:1.5 (1:(1~1.2):(1.5~5):(1~1.5)), with a reaction temperature of 0~60 °C and a reaction time of 12~36 h.

[0030] The optimal reaction molar ratio of compound C, N-Boc-4-aminopiperidine, to the base is 1:1.2:1.2~1:2:4 (1:(1.2~2):(1.2~4)), the optimal reaction temperature is 100~120 °C, and the optimal reaction time is 6~24 h.

[0031] The optimal reaction molar ratio of compound D, reducing agent (iron powder), and ammonium chloride aqueous solution is 1:2:2~1:10:10 (1:(2~10):(2~10)), the optimal reaction temperature is 60~85 °C, and the optimal reaction time is 8~24 h.

[0032] The optimal molar ratio of compound E to trifluoroacetic acid is 1:2 to 1:10 (1:(2 to 10)), the optimal reaction temperature is 0 to 25°C, and the optimal reaction time is 4 to 10 h.

[0033] The optimal reaction molar ratio of compound F, substituted benzyl chloride / benzyl bromide, and base is 1:1.1:1.2~1:1.3:2 (1:(1.1~1.3):(1.2~2)), the optimal reaction temperature is 25~45 °C, and the optimal reaction time is 4~24 h.

[0034] The optimal reaction molar ratio of compound II, aromatic boric acid / ester, Pd(dppf)Cl2, and base is 1:1.1:0.01:1.2~1:1.5:0.05:2 (1:(1.1~1.5):(0.01~0.05):(1.2~2)), with a reaction temperature of 100~120 °C and a reaction time of 12~24 h.

[0035] The present invention also provides a pharmaceutical composition comprising an effective dose of the above-described compound and a related pharmaceutical carrier.

[0036] The present invention also provides the use of the said compound or composition in the preparation of medicaments for the prevention and treatment of AIDS.

[0037] This invention, based on the binding mode of dihydropteridine-6(5H)-one skeleton compounds to HIV reverse transcriptase, and combined with computer-aided drug design, enhances the nonpolar interaction between the compounds and amino acid residues V179 and E138 in the binding pocket. Simultaneously, the left-wing structure penetrates deeper into the binding pocket to strengthen the binding force with highly conserved amino acid residues F227 and W229, further improving the bioactivity of the target compounds against drug-resistant HIV strains. In vitro cellular-level anti-HIV-1 activity experiments showed that this series of compounds exhibits significant anti-HIV-1 activity and low cytotoxicity. Detailed Implementation

[0038] The following examples can provide a better understanding of the invention, but they do not limit the scope of the invention.

[0039] Example 1: Synthesis of end product I

[0040] (1) Compound II (0.3 mmol, 1.0 eq), arylboronic acid or borate ester (0.36 mmol, 1.2 eq), Pd(dppf)Cl2 (0.015 mmol, 5 mol%) and cesium carbonate (0.6 mmol, 2.0 eq) were added to 1,4-dioxane / water (8 mL / 2 mL), the mixture was purged with nitrogen three times, and the temperature was raised to 110 °C under a nitrogen atmosphere. o The reaction was carried out at C for 12 h. After the reaction was confirmed to be complete by TLC, the reaction system was poured into 30 mL of water, extracted with ethyl acetate, dried and concentrated, and purified by column chromatography with dichloromethane / methanol as eluent to obtain a white solid.

[0041] (2) Compound II (0.4 mmol, 1.0 eq), arylboronic acid or borate ester (0.48 mmol, 1.2 eq), Pd(dppf)Cl2 (0.012 mmol, 3 mol%) and cesium carbonate (0.8 mmol, 2.0 eq) were added to 1,4-dioxane / water (8 mL / 2 mL), nitrogen gas was purged three times, and the mixture was heated to 110 °C under a nitrogen atmosphere. o The reaction was carried out at C for 12 h. After the reaction was confirmed to be complete by TLC, the reaction system was poured into 50 mL of water, extracted with ethyl acetate, dried and concentrated, and purified by column chromatography with dichloromethane / methanol as eluent to obtain a white solid.

[0042] (3) Compound II (0.5 mmol, 1.0 eq), arylboronic acid or borate ester (0.60 mmol, 1.2 eq), Pd(dppf)Cl2 (0.015 mmol, 3 mol%) and cesium carbonate (0.6 mmol, 2.0 eq) were added to 1,4-dioxane / water (10 mL / 2 mL), nitrogen gas was purged three times, and the mixture was heated to 110 °C under a nitrogen atmosphere. o The reaction was carried out at C for 12 h. After the reaction was confirmed to be complete by TLC, the reaction system was poured into 50 mL of water, extracted with ethyl acetate, dried and concentrated, and purified by column chromatography with dichloromethane / methanol as eluent to obtain a white solid.

[0043] The target compounds were obtained from starting materials containing different substituents using the above method. Some results are shown below:

[0044] (Ⅰa)

[0045] The operation method is the same as above. It is a white powdery solid with a yield of 64% and a maximum temperature of 268.5-280.9 °C. 1 H NMR (400MHz, DMSO-d6) δ 10.44 (s, 1H, NH), 7.95 (d, J = 8.0 Hz, 2H, ArH), 7.88 (d, J= 8.0 Hz, 2H, ArH), 7.73 (d, J = 8.1 Hz, 2H, ArH), 7.54 (s, 2H, ArH), 7.50(s, 1H, Pyr-H), 7.37 (d, J = 8.1 Hz, 2H, ArH), 7.30 (s, 2H, SO2NH2), 6.16 (s,1H, CONH), 4.16 (s, 2H, COCH2), 3.49-3.36 (m, 3H), 2.71-2.56 (m, 2H), 2.20(s, 6H, diMe), 1.92-1.53 ​​(m, 4H), 1.39-1.19 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 161.8, 158.5, 150.0, 144.7, 143.5, 143.1, 139.9, 139.7, 138.1, 137.7,133.4, 129.5, 128.1, 127.7, 126.1, 119.4, 110.6, 62.1, 52.9, 51.2, 48.8,32.0, 18.2. HRMS: m / z [M + H]+ calcd for [C 33 H 35 N8O3S] + , 623.2547, found623.2542.

[0046] (Ⅰb)

[0047] The operation method is the same as above. It is a white powdery solid with a yield of 81% and a mp of 223.9-225.2 °C. 1 H NMR (400MHz, DMSO-d6) δ 10.46 (s, 1H, NH), 9.10 (d, J = 2.3 Hz, 1H, Py-αH), 8.37 (dd,J = 8.2, 2.3 Hz, 1H, Py-γH), 8.01 (d, J = 8.2 Hz, 1H, Py-βH), 7.72 (d, J =7.9 Hz, 2H, ArH), 7.62 (s, 2H, ArH), 7.51 (s, 1H, Pyr-H), 7.44-7.33 (m, 2H,ArH), 7.30 (s, 2H, SO2NH2), 6.17 (s, 1H, CONH), 4.17 (s, 2H, COCH2), 3.62-3.41(m, 3H), 2.71-2.54 (m, 2H), 2.21 (s, 6H, diMe), 2.00-1.19(m, 6H). 13 C NMR (101MHz, DMSO-d6) δ 161.8, 158.5, 150.0, 148.7, 145.7 (q, J C-F = 33.9 Hz), 143.4,143.1, 140.06, 140.0, 138.9, 138.0, 136.5, 135.3, 129.5, 127.9, 126.1, 122.4(q, J C-F = 273.7 Hz), 121.4 (q, J C-F = 3.0 Hz), 110.5, 62.1, 52.9, 51.1, 48.8,31.9, 18.2. 19 F NMR (376 MHz, DMSO-d6) δ -66.17. HRMS: m / z [M + H] + calcd for[C 32 H34 F3N8O3S] + , 667.2421, found 667.2420.

[0048] (Ⅰc)

[0049] The operation method is the same as above. It is a white powdery solid with a yield of 43% and a maximum temperature of 272.2-274.3 °C. 1 H NMR (400MHz, DMSO-d6) δ 10.47 (s, 1H, NH), 7.93 (d, J = 7.8 Hz, 2H, ArH), 7.87 (d, J= 7.8 Hz, 2H, ArH), 7.69 (d, J = 7.1 Hz, 2H, ArH), 7.56-7.49 (m, 3H, Pyr-H,ArH), 7.46-7.37 (m, 3H, ArH, SO2NH), 6.19 (s, 1H, CONH), 4.17 (s, 2H, COCH2),3.60-3.43(m, 3H), 2.75-2.57 (m, 2H), 2.39 (d, J = 5.0 Hz, 3H, SO2NHCH3), 2.20(s, 6H, diMe), 2.00-1.53 ​​(m, 4H), 1.43-1.22 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 161.7, 158.5, 150.0, 144.7, 143.8, 139.8, 139.7, 138.3, 138.0, 137.7,133.4, 129.8, 128.0, 127.7, 127.1, 119.4, 110.5, 61.9, 52.9, 51.1, 48.5,31.8, 29.2, 18.2. HRMS: m / z [M + H] + calcd for [C 34 H 37 N8O3S] + , 637.2704, found637.2694.

[0050] (Ⅰd)

[0051] The operation method is the same as above. It is a white powdery solid with a yield of 40% and a maximum temperature of 210.4-212.0 °C. 1H NMR (400MHz, DMSO-d6) δ 10.45 (s, 1H), 8.01-7.84 (m, 5H), 7.79 (d, J = 7.8 Hz, 2H), 7.57-7.49 (m, 3H), 7.38-7.18 (m, 3H), 6.18 (s, 1H), 4.16 (s, 2H), 3.63-3.34(m, 3H), 2.77-2.59 (m, 2H), 2.20 (s, 6H), 1.97-1.22 (m, 6H). 13 C NMR (101 MHz, DMSO-d6) δ 168.2, 161.7, 158.4, 150.0, 144.7, 139.8, 139.6, 138.0, 137.6,133.6, 133.3, 129.1, 128.0, 127.9, 127.6, 119.3, 110.5, 61.9, 52.8, 51.1,48.7, 31.6, 18.2. HRMS: m / z [M + H] + calcd for [C 34 H 35 N8O2] + , 587.2877, found587.2873.

[0052] (Ⅰe)

[0053] The operation method is the same as above. It is a white powdery solid with a yield of 44% and a mp of 271.6-274.1 °C. 1H NMR (400MHz, CDCl3) δ 12.11 (s, 1H, NH), 7.77 (d, J = 8.0 Hz, 2H, ArH), 7.73-7.63 (m,4H, ArH), 7.56 (s, 1H, Pyr-H), 7.37 (s, 2H, ArH), 7.28 (s, 2H, ArH), 6.74 (s,1H, CONH), 6.17 (q, J = 4.8 Hz, 1H, CONHCH3), 4.33 (s, 2H, COCH2), 3.37 (s,2H, Ar-CH2-N), 3.23-3.09 (m, 1H, NHCH), 3.03 (d, J = 4.8 Hz, 3H, CONHCH3),2.72-2.60 (m, 2H), 2.28 (s, 6H, diMe), 1.87-1.70 (m, 4H), 1.48-1.37 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 168.1, 162.9, 158.9, 150.3, 145.1, 142.3, 139.1,138.6, 137.8, 137.3, 133.5, 132.7, 129.0, 127.8, 127.5, 126.8, 119.0, 111.2,109.4, 62.5, 50.9, 49.2, 31.6, 26.9, 18.1. HRMS: m / z [M + H] + calcd for[C 35 H 37 N8O2] + , 601.3034, found 601.3031.

[0054] (Ⅰf)

[0055] The operation method is the same as above. It is a white powdery solid with a yield of 59% and a maximum temperature of 162.3-164.8 °C. 1H NMR (400MHz, CDCl3) δ 12.57 (s, 1H, NH), 8.10 (d, J = 8.2 Hz, 2H, ArH), 7.76 (d, J =8.1 Hz, 2H, ArH), 7.69 (d, J = 8.1 Hz, 2H, ArH), 7.59 (s, 1H, Pyr-H), 7.43-7.31 (m, 4H, ArH), 7.10 (br. s, 1H, CONH), 4.32 (s, 2H, COCH2), 3.39 (s, 2H,Ar-CH2-N), 3.18-3.03 (m, 1H, CHNH), 2.63 (d, J = 10.7 Hz, 2H, N(CH2CH2)2), 2.26 (s, 6H, diMe), 2.09-1.72 (m, 4H, N(CH2CH2)2), 1.43 (dd, J = 12.3, 11.6Hz, 2H, N(CH2CH2)2). 13 C NMR (101 MHz, CDCl3) δ 162.9, 158.7, 150.4, 147.1,146.7, 145.0, 139.1, 138.5, 137.2, 132.7, 129.3, 128.4, 127.7, 127.5, 123.5,118.8, 111.3, 109.4, 62.0, 52.4, 50.9, 49.0, 31.5, 18.1. HRMS: m / z [M + H] + calcd for [C 33 H 33 N8O3] + , 589.2670, found 589.2677.

[0056] (Ⅰg)

[0057] The operation method is the same as above. It is a white powdery solid with a yield of 61% and a mp of 196.8-198.8 °C. 1H NMR (400MHz, CDCl3) δ 12.55 (s, 1H, NH), 8.47 (d, J = 5.4 Hz, 2H, Py-αH), 7.75 (d, J= 8.1 Hz, 2H, ArH), 7.67 (d, J = 8.1 Hz, 2H, ArH), 7.55 (s, 1H, Pyr-H), 7.36(s, 2H, ArH), 7.14 (d, J = 5.4 Hz, 2H, Py-βH), 7.00 (s, 1H, CONH), 4.32 (s,2H, COCH2), 3.30 (s, 2H, Ar-CH2-N), 3.18-3.03 (m, 1H, NHCH), 2.62 (d, J = 10.7Hz, 2H), 2.26 (s, 6H, diMe), 2.07-1.93 (m, 2H), 1.84 -1.72 (m, 2H), 1.49-1.37(m, 2H). 13 C NMR (101 MHz, CDCl3) δ 163.0, 158.8, 150.3, 149.6, 148.0, 145.0,139.0, 138.5, 137.6, 137.3, 132.6, 127.7, 127.4, 123.7, 118.8, 111.2, 109.3,61.7, 52.4, 50.8, 49.0, 31.5, 18.1. HRMS: m / z [M + H] + calcd for [C 32 H 33 N8O] + ,545.2772, found 545.2774.

[0058] (Ⅰh)

[0059] The operating method is the same as above. It is a white powdery solid with a yield of 54% and a maximum temperature of 180.3-181.5 °C. 1H NMR (400MHz, DMSO-d6) δ 10.46 (s, 1H, NH), 7.93 (d, J = 8.2 Hz, 2H, ArH), 7.87 (d, J= 8.2 Hz, 2H, ArH), 7.52 (s, 3H, ArH, Pyr-H), 7.40 (d, J = 7.8 Hz, 2H, ArH),7.22 (d, J = 7.8 Hz, 2H, ArH), 6.14 (s, 1H, CONH), 4.16 (s, 2H, CONH2), 3.41-3.23 (m, 7H), 2.71-2.58 (m, 2H), 2.19 (s, 6H, diMe), 1.89-1.19 (m, 10H). 13 CNMR (101 MHz, DMSO-d6) δ 168.6, 161.7, 158.5, 150.0, 144.7, 140.6, 139.9,139.7, 138.0, 137.7, 136.2, 133.3, 128.8, 128.0, 127.6, 127.4, 119.3, 110.5,62.2, 52.8, 51.1, 49.4, 48.8, 46.4, 31.9, 26.4, 24.4, 18.2. HRMS: m / z [M + H] + calcd for [C 38 H 41 N8O2] + , 641.3347, found 641.3347.

[0060] (Ⅰi)

[0061] The operation method is the same as above. It is a white powdery solid with a yield of 68% and a maximum temperature of 224.2-225.6 °C. 1H NMR (400MHz, DMSO-d6) δ 10.47 (s, 1H, NH), 9.08 (t, J = 6.0 Hz, 1H, Ar-CONH), 7.93 (d, J = 8.1 Hz, 2H, ArH), 7.86 (d, J = 8.1 Hz, 2H, ArH), 7.80 (d, J = 7.9 Hz,2H, ArH), 7.54 (s, 1H, Pyr-H), 7.52 (s, 2H, ArH), 7.35 (d, J = 5.1 Hz, 1H,thiophene-αH), 7.27 (d, J = 7.9 Hz, 2H, ArH), 7.01 (d, J = 3.4 Hz, 1H,thiophene-β'H), 6.94 (dd, J = 5.1, 3.4 Hz, 1H, thiophene-βH), 6.14 (s, 1H,CONH), 4.63 (d, J = 6.0 Hz, 2H, Ar-CONH-CH2-thiophene), 4.17 (s, 2H, COCH2),3.40-3.23 (m, 3H), 2.71-2.56 (m, 2H), 1.97-1.18 (m, 6H). 13 C NMR (101 MHz, DMSO-d6) δ 166.4, 161.7, 158.5, 150.0, 144.7, 143.2, 142.6, 139.9, 139.6, 138.0, 137.7, 133.3, 133.2, 129.0, 128.0, 127.6, 127.1, 125.8, 125.3, 119.3,110.5, 62.2, 52.8, 51.1, 48.7, 38.2, 31.9, 18.2. HRMS: m / z [M + H] + calcd for[C 39 H 39 N8O2S] + , 683.2911, found 683.2908.

[0062] Example 2: Anti-HIV bioactivity test

[0063] The in vitro cellular anti-HIV activity was determined by the Rega Institute of Pharmaceutical Research at the University of Katholleke, 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 concentration required to protect 50% of the cells from HIV-induced cytopathic effects was calculated as the half-effective concentration (EC50). 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 that caused cytopathic effects in 50% of uninfected cells (Half-Cytotoxicity concentration, CC). 50 And calculate the selectivity index (SI), SI = CC 50 / EC 50 .

[0064] Materials and Methods:

[0065] The anti-HIV bioactivity of each compound was monitored by the efficiency of its inhibitory effect on HIV-induced cytopathic effects in cells. MT-4 cells were used for cell culture. The viral strain used was HIV-1 strain IIIB.

[0066] 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 pre-cultured at 37 °C for 1 h with 100 μL of solutions containing different concentrations of various compounds. Then, 100 μL of an appropriate viral dilution was added to each compound, and the cells were incubated at 37 °C for 1 h. After three washes, the cells were resuspended in culture media containing or without the compounds. The cells were then cultured at 37 °C for 7 days in a 5% CO2 atmosphere, with the culture medium being replaced with either compound-containing or compound-free media 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 viral dilutions 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 viral cytopathic effects without direct cytotoxicity to cells (CC). 50It 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, comparative experiments on antiviral activity using a blank sample 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.

[0067] This invention uses marketed drugs nevirapine (NVP), efavirenz (EFV), and ETR as reference standards. The results of the inhibitory activity of some target compounds against HIV are shown in Table 1.

[0068] ;

[0069] Table 1

[0070] .

[0071] a EC 50 The effective concentration that protects 50% of cells from viral infection; b RES056 represents the K103N / Y181C double mutant strain;

[0072] c L represents compound EC 50 Values ​​range from 1 to 10 nM.

[0073] The present invention is not limited to the above examples.

Claims

1. A dihydropteridine-6(5H)-one skeleton compound, characterized in that, The structural formula is as follows: ； Among them, R 1 Selected from cyano, cyanovinyl, various substituted or unsubstituted benzene rings, pyridine, pyrimidine, azines and their oxides, thiophene (furan) (sulfoxide and sulfone), (iso)thiazole and its oxides, pyrrole and its oxides, pyrazole (furan) and its oxides, imidazole and its oxides, (iso)oxazole and its oxides, pyrazolone, furan, cyclopentadiene, dihydrothiophene and its cyclic sulfides (sulfoxide and sulfone), dihydrofuran and its epoxides, and cycloalkanes; R 2 Selected from H, CN, NO2, OH, CH2OH, CF3, F, Cl, Br, CH3, COOMe, COOH, SO2CH3, SONH2, CONH2 and a series of sulfonyl (derivatives) and amide (derivatives) with hydrophilic or lipophilic ends, with substituents being ortho, meta, para monosubstituted or polysubstituted; X is a linker among -H-, -N-, -CH-, -S-, -S(O)-, and -S(O)2-.

2. The dihydropteridine-6(5H)-one skeleton compound according to claim 1, characterized in that, It also includes salts, stereochemical isomers, hydrates or solvates of the compound.

3. The dihydropteridine-6(5H)-one skeleton compound according to claim 2, characterized in that, The salt is a sodium salt, hydrochloride, hydrobromide, sulfate, formate, acetate, phosphate, tartrate or citrate, succinate, maleate, trifluoromethanesulfonate, p-toluenesulfonate, methanesulfonate, fumarate or malate.

4. The method for preparing the dihydropteridine-6(5H)-one skeleton compound as described in claim 1, characterized in that, The general reaction formula is: ； The specific preparation steps are as follows: (i) In a solvent, using 2,6-dimethyl-4-bromoaniline (A) and 2,4-dichloro-5-nitro-pyrimidine (B) as raw materials, hydrogen is abstracted under the action of a base, and then reacted with ethyl bromoacetate to obtain compound C; (ii) Subsequently, compound C reacts with N-Boc-4-aminopiperidine under appropriate solvent and alkaline conditions to give compound D; (iii) After separation, compound D undergoes a cyclization reaction under conditions of reduced iron powder and ammonium chloride aqueous solution to obtain compound E; (iv) Compound E was deprotected from its Boc protecting group in a mixed solvent of trifluoroacetic acid and dichloromethane to give compound F; (v) Compound F reacts with benzyl bromide or benzyl chloride with different functional groups in a solvent under the catalysis of a base to give compound II; (vi) Compound II then undergoes a metal coupling reaction with an aromatic boric acid or ester to obtain the target product, dihydropteridine-6(5H)-one skeleton compound I.

5. The preparation method according to claim 4, characterized in that... ； The solvent used in the entire reaction is one or more of N,N-dimethylformamide, N,N-dimethylacetamide, 1,4-dioxane, acetone, acetonitrile, toluene, dichloromethane, tetrahydrofuran, methanol, ethanol, isopropanol, n-butanol, and isobutanol, or there may be no solvent reaction. The base used is one or more of the following: sodium carbonate, sodium bicarbonate, potassium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide, sodium hydride, N,N-dimethylaminopyridine, triethylamine, diisopropylethylamine, tributylamine, potassium tert-butoxide, and sodium tert-butoxide. The acid used is one or more of the following: hydrochloric acid, sulfuric acid, trifluoroacetic acid, formic acid, acetic acid, and ammonium chloride aqueous solution.

6. The preparation method according to claim 4, characterized in that... ； The molar ratio of compounds A, B, ethyl bromoacetate, and base is 1:1:1.5:1 to 1:1.2:5:1.5, the reaction temperature is 0 to 60°C, and the reaction time is 12 to 36 h. The molar ratio of compound C, N-Boc-4-aminopiperidine, and base is 1:1.2:1.2~1:2:4, the reaction temperature is 100~120 °C, and the reaction time is 6~24 h; The molar ratio of compound D, iron powder, and ammonium chloride is 1:2:2~1:10:10, the reaction temperature is 60~85 °C, and the reaction time is 8~24 h; The molar ratio of compound E to trifluoroacetic acid is 1:2 to 1:10, the reaction temperature is 0 to 25 °C, and the reaction time is 4 to 10 h. The molar ratio of compound F, substituted benzyl chloride / benzyl bromide, and base is 1:1.1:1.2 to 1:1.3:2, the reaction temperature is 25 to 45°C, and the reaction time is 4 to 24 h. The molar ratio of compound II, aromatic boric acid / ester, Pd(dppf)Cl2, and base was 1:1.1:0.01:1.2 to 1:1.5:0.05:2, the reaction temperature was 100 to 120 °C, and the reaction time was 12 to 24 h.

7. A pharmaceutical composition comprising an effective dose of the dihydropteridine-6(5H)-one skeleton compound as described in claim 1, or a salt thereof, or a stereochemical isomer thereof, or a hydrate or solvate thereof, and a pharmaceutical carrier.

8. The use of the dihydropteridine-6(5H)-one skeleton compound as described in claim 1, or its pharmaceutical salt, its stereochemical isomer, its hydrate or solvate, in the preparation of medicaments for the prevention and treatment of AIDS.

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