CN108586482A - A kind of Diarylmiazines HIV-1 inhibitor of the ring containing triazole and its preparation method and application - Google Patents

Abstract

The invention relates to a diarylpyrimidine HIV‑1 inhibitor containing a triazole ring, a preparation method and application thereof. The compound has the structure of formula I. The present invention also relates to a pharmaceutical composition containing the compound of formula I. The present invention also provides the application of the above-mentioned compounds and compositions containing one or more such compounds in the preparation of medicines for treating and preventing human immunodeficiency virus (HIV).

Description

A kind of diarylpyrimidine HIV-1 inhibitor containing triazole ring and preparation method thereof and application

technical field

The invention belongs to the technical field of organic compound synthesis and medical application, in particular to a diarylpyrimidine HIV-1 inhibitor containing a triazole ring and its preparation method and application.

Background technique

AIDS, also known as acquired immunodeficiency syndrome (Acquired Immunodeficiency Syndrome, AIDS), is a very harmful infectious disease caused by human immunodeficiency virus (human immunodeficiency virus, HIV) infection. After an infected person is infected, the body's immune barrier is usually severely damaged, making it easy to suffer from various opportunistic infections and tumors, as well as chronic diseases. Thus AIDS has become one of the chronic diseases with high mortality rate.

HIV-1 reverse transcriptase (reverse transcriptase, RT) plays a key role in the replication cycle of the virus, making it an important target for the development of anti-HIV-1 drugs. Inhibitors acting on RT are mainly divided into nucleoside reverse transcriptase inhibitors (Nucleoside Reverse Transcriptase Inhibitors, NRTIs) and non-nucleoside reverse transcriptase inhibitors (Non-nucleoside Reverse Transcriptase inhibitors, NNRTIs). Among them, NNRTIs are an important part of the highly active antiretroviral therapy (HAART) for the treatment of AIDS due to their high activity, strong selectivity, low toxicity and many other advantages. However, the problems of drug resistance, toxic side effects and poor pharmacokinetic properties of NNRTIs in clinical treatment limit their clinical application to a certain extent. Therefore, the research and development of novel NNRTIs with high efficiency and low toxicity, broad-spectrum anti-drug resistance and good pharmacokinetic properties is one of the important directions of current anti-AIDS drug research.

Contents of the invention

Aiming at the deficiencies of the prior art, the present invention provides a diarylpyrimidine HIV-1 inhibitor containing a triazole ring and a preparation method thereof, and the present invention also provides the activity screening results of the above compound as an HIV-1 inhibitor and its application.

Technical scheme of the present invention is as follows:

1. Diarylpyrimidine HIV-1 inhibitors containing a triazole ring

A diarylpyrimidine HIV-1 inhibitor containing a triazole ring, or a pharmaceutically acceptable salt thereof, has a structure shown in general formula I:

in,

X is: O or MH;

R is: CH ₃ , CN or CH=CHCN;

R ¹ is: phenyl or pyridyl; or halogen, SO ₂ NH ₂ , SO ₂ CH ₃ , CONH ₂ , NO ₂ , CN, NH ₂ , CF ₃ , NHCH ₃ , OH, COOH, CH ₂ OH, CO ₂ Me, OCH ₃ , NHCOCH ₃ substituted phenyl; the substituent is ortho, meta, para mono-substituted or multi-substituted; or CONR ² ;

Wherein, R ² is: phenyl; or halogen, SO ₂ NH ₂ , SO ₂ CH ₃ , CONH ₂ , NO ₂ , CN, NH ₂ , CF ₃ , NHCH ₃ , OH, COOH, CH ₂ OH, CO ₂ Me , OCH ₃ , NHCOCH ₃ substituted phenyl; the substituents are ortho, meta, para mono-substituted or multi-substituted; five-membered heterocycle, six-membered heterocycle.

Preferably according to the present invention, the diarylpyrimidine HIV-1 inhibitor containing a triazole ring is one of the following compounds:

The "pharmaceutically acceptable salt" mentioned in the present invention means that within the scope of reliable medical evaluation, the salt of the compound is suitable for contacting with human or lower animal tissues without undue toxicity, irritation and allergy reactions, etc., have a fairly reasonable ratio of benefit to risk, are usually water or oil soluble or dispersible, and are effective for their intended use. Included are the pharmaceutically acceptable acid addition salts and the pharmaceutically acceptable base addition salts, which are acceptable for the intended use and chemically compatible with the compounds of formula I herein. For a list of suitable salts see S.M. Birge et al., J. Pharm. Sci., 1977, 66, pp. 1-19.

2. The preparation method of the diarylpyrimidine HIV-1 inhibitor containing triazole ring

The preparation method of a diarylpyrimidine HIV-1 inhibitor containing a triazole ring comprises the following steps: using 2,4-dichloro-substituted thieno[3,2-d]pyrimidine 1 as an initial raw material, and first , N-dimethylformamide solution reacts with substituted phenol or aniline through nucleophilic substitution to generate intermediate 2; then intermediate 2 reacts with N-Boc-4-aminopiperidine to generate intermediate 3 through nucleophilic substitution, and then Deprotection of Boc protection in trifluoroacetic acid affords intermediate 4. Then 4 reacts with propyne bromide to generate the key intermediate 5, and then undergoes a copper-catalyzed azide-terminal alkyne cycloaddition reaction (CuAAC Click) reaction with the azide substituent to obtain the target product compound of formula I. The synthetic route is as follows:

Reagents and conditions: (i) substituted phenol or aniline, N,N-dimethylformamide, potassium carbonate, room temperature; (ii) N-Boc-4-aminopiperidine, N,N-dimethylformamide, Potassium carbonate, 110°C; (iii) dichloromethane, trifluoroacetic acid, room temperature; (iv) propyne bromide, N,N-dimethylformamide, potassium carbonate, room temperature; (v) sodium vitamin C, pentahydrate Copper sulfate, N,N-dimethylformamide, water, azide substituent, 50°C;

X, R, R ¹ are shown in the same general formula I above;

The substituted phenols or anilines are: s-trimethylphenol, 2,6-dimethyl-4-cyanophenol, 2,6-dimethyl-4-(E)-cyanovinylphenol, Trimethylaniline, 2,6-dimethyl-4-cyanoaniline, 2,6-dimethyl-4-(E)-cyanovinylaniline;

The azide substituents are: o-chlorobenzyl azide, m-chlorobenzyl azide, p-chlorobenzyl azide, o-bromobenzyl azide, m-bromobenzyl azide, p-bromobenzyl azide , o-fluorobenzyl azide, m-fluorobenzyl azide, p-fluorobenzyl azide, 2,4-difluorobenzyl azide, 3,4-difluorobenzyl azide, o-cyanobenzyl azide Nitrogen, m-cyanobenzyl azide, p-cyanobenzyl azide, o-nitrobenzyl azide, m-nitrobenzyl azide, p-nitrobenzyl azide, o-methoxybenzyl azide , m-methoxybenzyl azide, p-methoxybenzyl azide, p-methylsulfonyl benzyl azide, m-methylsulfonyl benzyl azide, o-methylsulfonyl benzyl azide, p-sulfonyl benzyl azide Benzyl azide, m-sulfonamidobenzyl azide, o-sulfonamidobenzyl azide, p-carboxamidobenzyl azide, m-carboxamidobenzyl azide, o-carboxamidobenzyl benzyl azide , 2-azido-1-(pyrrolidin-1-yl)ethan-1-one, 2-azido-N-cyclopropylacetamide, 2-azido-1-morpholine-1- ketone.

The room temperature described in the present invention is 20-30°C.

3. Anti-HIV-1 activity and application of diarylpyrimidine HIV-1 inhibitors containing triazole ring

The present invention has carried out anti-HIV-1 (III _B ) at the cell level on the part of diarylpyrimidine derivatives containing triazole rings synthesized according to the above-mentioned method, and screened the activity of double drug-resistant mutant strain RES056 (K103N/Y181C) , with nevirapine (NVP) and etravirine (ETV) as positive controls.

The activity results are shown in Table 1. Most of the compounds exhibited strong anti-HIV-1IIIB activity, with EC50 values in the range of 3.28-10.4nM, far superior to the positive drug NVP (EC50=163nM). Among them, compounds A7 (EC50=3.28nM) and A9 (EC50=4.38nM) showed the best anti-HIV-1IIIB activity, better than the latest generation drug ETV (EC50=5.1nM). And A7 (CC50>210μM, SI>64103) and A9 (CC50>212μM, SI>48544) both have extremely low cytotoxicity, showing extremely high selectivity.

The diaryl pyrimidine HIV-1 inhibitor containing a triazole ring of the invention can be used as a non-nucleoside HIV-1 inhibitor. Specifically, it is used as an HIV-1 inhibitor for the preparation of anti-AIDS drugs.

An anti-HIV-1 pharmaceutical composition, comprising the triazole ring-containing diarylpyrimidine HIV-1 inhibitor of the present invention and one or more pharmaceutically acceptable carriers or excipients.

The present invention provides a novel biarylpyrimidine HIV-1 inhibitor containing a triazole ring and its preparation method. The present invention also provides the screening results of the anti-HIV-1 activity of the compound and its first antiviral activity in the antiviral field. application. Tests have proved that the triazole ring-containing diarylpyrimidine derivatives of the present invention can be used as HIV-1 inhibitors and have high application value. Specifically, it is used as an HIV-1 inhibitor for the preparation of anti-AIDS drugs.

Detailed ways

The following examples help to understand the present invention, but the content of the present invention cannot be limited.

The synthetic route involved in the embodiment is as follows:

Example 1: Preparation of 4-((2-chlorothieno[3,2-d]pyrimidin-4-yl)oxy)-3,5-dimethylbenzonitrile (6)

Weigh 4-hydroxy-3,5-dimethylbenzonitrile (1.5g, 10mmol) and potassium carbonate (1.7g, 12mmol) in 30mL of DMF, stir at room temperature for 15 minutes, then add 2,4-dichlorothiophene And[3,2-d]pyrimidine 1 (1.9g, 10mmol) continued to stir at room temperature for 2h (TLC detected that the reaction was complete). After a large amount of white solid was formed, 100 mL of ice water was slowly added thereto, filtered, dried in vacuo, and recrystallized from ethanol to obtain Intermediate 6. White solid, yield 93.8%, melting point 258-260°C. ESI-MS: m/z316.3[M+1] ⁺ .C ₁₅ H ₁₀ ClN ₃ OS(315.02).

Example 2: 3,5-Dimethyl-4-((2-(piperidin-4-ylamino)thieno[3,2-d]pyrimidin-4-yl)oxy)benzonitrile (8) preparation of

6 (0.95g, 3.17mmol), N-Boc-4-aminopiperidine (0.83g, 3.80mmol) and potassium carbonate (0.87g, 6.33mmol) were added to 20mL of DMF in turn, then heated to reflux for 10h (TLC detection). After the reaction was cooled to room temperature, the reaction solution was slowly added dropwise to 50 mL of water, and a large amount of yellow solid was formed. After standing for 30min, filter and vacuum dry to obtain the crude product. The crude product (1.26g, 2.53mmol) was weighed and dissolved in 4mL of dichloromethane, 2.22mL of trifluoroacetic acid (30mmol) was added, and stirred at room temperature for 3-5h (TLC detection). Then the pH of the reaction solution was adjusted to 9 with saturated sodium bicarbonate solution, extracted with dichloromethane (3×5 mL), washed with saturated sodium chloride solution, and the organic layer was dried over anhydrous sodium sulfate. Intermediate 8 was then isolated by flash column chromatography. White solid, yield 84.2%, melting point: 114-116°C.1H NMR (400MHz, DMSO-d6, ppm) δ: 8.20(d, J=5.4Hz, 1H), 7.72(s, 1H), 7.26(s ,1H),6.92(s,1H),3.78(s,1H), 2.89(s,2H),2.12(s,6H),1.74-1.78(m,2H),1.23-1.28(m,4H). ESI-MS: m/z 380.5[M+1] ⁺ . C ₂₀ H ₂₁ N ₅ OS(379.15).

Example 3: 3,5-Dimethyl-4-((2-((1-(prop-2-yn-1-yl)piperidin-4-yl)amino)thieno[3,2-d Preparation of ]pyrimidin-4-yl)oxy)benzonitrile (9)

Weigh intermediate 8 (0.5mmol, 0.19g) in 5mL DMF, then add anhydrous potassium carbonate (1.0mmol, 0.14g) and bromide propyne (0.6mmol, 0.07g) successively, react under room temperature (TLC detection The reaction is complete). After the reaction was complete, 20 mL of saturated brine was added to the reaction liquid, washed with ethyl acetate (3×15 mL), and the organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Separation by flash column chromatography, and then recrystallization in ethyl acetate-petroleum ether gave the target compound 9. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.20 (d, J=5.3Hz, 1H, C ₆ -thienopyrimidine-H), 7.72 (s, 2H, C ₃ , C ₅ -Ph-H), 7.27 (s,1H,C ₇ -thienopyrimidine-H),6.88(s,1H,NH),3.69(s,1H),3.22(s,2H,N-CH ₂ ),3.13(s,1H,CH≡) ,2.74(s,2H),2.12(s,6H),1.90–1.30(m,6H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ162.4,160.5,153.4,133.2,132.9,123.7,119.0,109.0 ,75.9,51.3,46.6,31.5,16.2. ESI-MS: m/z 418.5[M+1] ⁺ .C ₂₃ H ₂₃ N ₅ OS(417.16).

Embodiment 4: the preparation of target compound A1-A10

9 (0.5 mmol) and different azide substituents (0.6 mmol) were added to DMF/H ₂ O mixed solvent (v/v=1:1, 10 mL). Then, 1 M freshly prepared aqueous solution of sodium vitamin C (0.1 mmol) and 7.5% aqueous solution of CuSO ₄ ·5H ₂ O (0.1 mmol) were added to the mixed solution. Heated to 50°C and stirred for 4-12h (TLC detection). After the reaction, a large amount of precipitate was formed, which was filtered, and the filter cake was washed with water and dried. Then recrystallized in methanol solution to obtain the target products A1-A10.

4-((2-((1-((1-(2-cyanobenzyl)-1H-1,2,3-triazol-4-yl)methyl)piperidin-4-yl)amino) Thieno[3,2-d]pyrimidin-4-yl)oxy)-3,5-dimethylbenzonitrile (A1)

White solid, yield 92%, melting point 149-151°C. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.20 (d, J=5.4Hz, 1H, C ₆ -thienopyrimidine-H), 8.08 (s, 1H, triazol-H), 7.92 (dd, J=7.7 ,1.4Hz,1H,C ₃ -Ph'-H),7.73 (d,J＝1.5Hz,1H,C ₅ -Ph'-H),7.72(s,2H,C ₃ ,C ₅ -Ph"- H),7.60–7.55(m,1H,C ₆ -Ph'-H),7.36(d, J＝7.8Hz,1H,C ₄ -Ph'-H),7.26–7.25(m,1H,C ₇ -thienopyrimidine-H),6.88(s,1H,NH),5.80(s,2H,triazol-CH ₂ ),3.71(s,1H),3.56(s,2H,N-CH ₂ ),2.80(s, 2H),2.11(s,6H),1.72-1.53(m,3H), 1.45–1.39(m,3H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ162.4,153.4,139.4,134.3,133.8,133.2 , 133.0,129.8,129.6,124.9,119.0,117.4,111.6,109.0,52.9,52.3,51.4,39.6,31.5,16.2. ESI-MS: m/z 576.6[M+1] ⁺ .C ₃₁ H ₂₉ N ₉ OS(575.22).

4-((2-((1-((1-(4-cyanobenzyl)-1H-1,2,3-triazol-4-yl)methyl)piperidin-4-yl)amino) Thieno[3,2-d]pyrimidin-4-yl)oxy)-3,5-dimethylbenzonitrile (A2)

White solid, yield 90%, melting point 140-142°C. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.20(d, J=5.3Hz, 1H, C ₆ -thienopyrimidine-H), 8.13(s, 1H, triazol-H), 7.86(d, J=8.0 Hz, 2H, C ₃ , C ₅ -Ph'-H), 7.72(s, 2H, C ₃ , C ₅ -Ph"-H), 7.44(d, J＝7.5Hz, 2H, C ₂ , C ₆ -Ph'-H), 7.27(s,1H,C ₇ -thienopyrimidine-H), 6.87(s,1H,NH),5.73(s,2H,triazol-CH ₂ ),3.72–3.71(s,1H) ,3.54(s,2H,N-CH ₂ ),2.79(s,2H), 2.11(s,6H),1.91–1.57(m,4H),1.45–1.39(s,2H). ¹³ CNMR(100MHz, DMSO-d ₆ )δ162.4, 153.4, 139.4, 134.3, 133.8, 133.2, 133.0, 129.8, 129.6, 124.9, 119.0, 117.4, 111.6, 109.0, 52.9, 52.3, 51.4, 39.6, 31.5, 16-ESI.2. :m/z 576.6[M+1] ⁺ .C ₃₁ H ₂₉ N ₉ OS(575.22).

3,5-Dimethyl-((2-((1-((1-(2-nitrobenzyl)-1H-1,2,3-triazol-4-yl)methyl)piperidine- 4-yl)amino)thieno[3,2-d]pyrimidin-4-yl)oxy)benzonitrile (A3)

White solid, yield 93%, melting point 155-157°C. ¹ H NMR (400MHz,DMSO-d ₆ )δ8.21(d,J＝5.4Hz, 1H,C ₆ -thienopyrimidine-H),8.19–8.17(m,1H),8.01(s,1H,triazol-H ),7.72(s,2H,C ₃ ,C ₅ -Ph”-H),7.62–7.55(m,2H),7.32–7.31(m,2H),7.26–7.24(m,1H,C ₇ -thienopyrimidine -H),6.88(s,1H,NH),5.80(s,2H,triazol-CH ₂ ),3.70(s,1H),3.52(s,2H,N-CH ₂ ),2.80(s,2H) ,2.11(s,6H),1.85-1.60(m,4H),1.42–1.31(m,2H). ¹³ C NMR(100MHz, DMSO-d ₆ )δ162.4,153.4,139.7,134.3,133.8,133.2,133.0 ,129.8,124.9,119.0,117.4,111.6, 109.0,52.9,52.3,51.3,39.6,31.5,16.2. ESI-MS: m/z 596.4[M+1] ⁺ .C30H29N9O3S(595.21).

3,5-Dimethyl-((2-((1-((1-(2-methylbenzyl)-1H-1,2,3-triazol-4-yl)methyl)piperidine- 4-yl)amino)thieno[3,2-d]pyrimidin-4-yl)oxy)benzonitrile (A4)

White solid, yield 88%, melting point 114-116°C. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.20 (d, J=5.4Hz, 1H, C ₆ -thienopyrimidine-H), 7.91 (s, 1H, triazol-H), 7.71 (s, 2H, C ₃ ,C ₅ -Ph”-H), 7.28–7.18(m, 4H), 7.05(d, J=7.4Hz, 1H), 6.87(s, 1H, NH), 5.58(s, 2H, triazol-CH ₂ ),3.68(s,1H),3.52(s,2H, N-CH ₂ ),2.77(s,2H),2.30(s,3H),2.11(s,6H),1.90–1.39(m,6H ). ¹³ C NMR (100MHz, DMSO-d ₆ ) δ162.4, 153.4, 136.6, 134.8, 133.2, 130.8, 128.9, 128.7, 126.7, 124.9, 119.0, 117.4, 111.6, 109.0, 53.0, 52.3, 51.3, 19 ,19.1,16.2.ESI-MS: m/z 565.6[M+1] ⁺ .C ₃₁ H ₃₂ N ₈ OS (564.24).

3,5-Dimethyl-((2-((1-((1-(4-methylbenzyl)-1H-1,2,3-triazol-4-yl)methyl)piperidine- 4-yl)amino)thieno[3,2-d]pyrimidin-4-yl)oxy)benzonitrile (A5)

White solid, yield 84%, melting point 112-114°C. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.20 (d, J=5.4Hz, 1H, C ₆ -thienopyrimidine-H), 7.98 (s, 1H, triazol-H), 7.71 (s, 2H, C ₃ ,C ₅ -Ph”-H),7.25–7.12(m, 5H),6.87(s,1H,NH),5.51(s,2H,triazol-CH ₂ ),3.68(s,1H),3.50( s,2H,N-CH ₂ ),2.76(s,2H), 2.28(s,3H),2.11(s,6H),1.87–1.62(m,3H),1.50–1.39(m,3H). ¹³ C NMR(100MHz, DMSO-d ₆ )δ162.4,153.4,139.4,137.8,134.3,133.6,133.2,129.7,128.3,124.9,119.0,117.8,109.0,52.9,52.3,31.5,21.1,16.2.ESI-MS m/z 565.6[M+1] ⁺ .C ₃₁ H ₃₂ N ₈ OS(564.24).

4-((4-((4-((4-(4-cyano-2,6-dimethylphenoxy)thieno[3,2-d]pyrimidin-2-yl)amino)piperidine -1-yl)methyl)-1H-1,2,3-triazol-1-yl)methyl)benzenesulfonamide (A6)

White solid, yield 93%, melting point 186-188°C. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.20(d, J=5.4Hz, 1H, C ₆ -thienopyrimidine-H), 8.07(s, 1H, triazol-H), 7.82(d, J=8.2 Hz, 2H, C ₃ , C ₅ -Ph'-H), 7.72 (s, 2H, C ₃ , C ₅ -Ph"-H), 7.46 (d, J＝8.0Hz, 2H, C ₂ , C ₆ -Ph'-H), 7.38(s,2H,SO ₂ NH ₂ ),7.26(s,1H,C ₇ -thienopyrimidine-H),6.87(s,1H,NH),5.67(s,2H,triazol- CH ₂ ),3.69(s,1H),3.52(s,2H, N-CH ₂ ),2.76(s,2H),2.11(s,6H),1.89–1.52(m,4H),1.46–1.22( m,2H). ¹³ C NMR(100MHz, DMSO-d ₆ )δ162.4,153.4,144.5,144.2,140.4,139.4,133.2,133.0,129.8,128.7,126.5,124.5, 119.0,117.6,112.1,109.1,0,5 52.5, 52.4, 39.6, 31.6, 16.2. ESI-MS: m/z 530.5[M+1] ⁺ . C ₃₀ H ₃₁ N ₉ O ₃ S ₂ (629.20).

4-((4-((4-((4-(4-cyano-2,6-dimethylphenoxy)thieno[3,2-d]pyrimidin-2-yl)amino)piperidine -1-yl)methyl)-1H-1,2,3-triazol-1-yl)methyl)benzamide (A7)

White solid, yield 91%, melting point 200-202°C. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.20(d, J=5.4Hz, 1H, C ₆ -thienopyrimidine-H), 8.05(s, 1H, triazol-H), 7.86(d, J=8.0 Hz, 2H, C ₂ , C ₆ -Ph'-H), 7.71(s, 2H, C ₃ , C ₅ -Ph"-H), 7.40(s, 1H, CONH ₂ ), 7.34(d, J＝ 8.0Hz,2H,C ₃ ,C ₅ -Ph'-H),7.27(s,1H,C ₇ -thienopyrimidine-H),6.87(s,1H,NH),5.63(s,2H,triazol-CH ₂ ),3.69(s,1H),3.51(s,2H, N-CH ₂ ),2.77(s,2H),2.11(s,6H),1.91–1.72(m,3H),1.51–1.23(m, 3H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ167.8, 165.4, 162.8, 153.1, 144.5, 139.7, 134.4, 133.2, 133.0, 128.3, 128.0, 126.1, 124.4, 119.0, 117.1, 112.8, 5.109 52.7, 52.4, 39.6, 31.6, 16.2. ESI-MS: m/z 594.6[M+1] ⁺ . C ₃₁ H ₃₁ N ₉ O ₂ S (593.23).

2-(4-((4-((4-(4-cyano-2,6-dimethylphenoxy)thieno[3,2-d]pyrimidin-2-yl)amino)piperidine- 1-yl)methyl)-1H-1,2,3-triazol-1-yl)-N-(4-(methylsulfonyl)phenyl)acetamide (A8)

White solid, yield 93%, melting point 173-175°C. ¹ H NMR (400MHz,DMSO-d ₆ )δ10.93(s,1H,CONH),8.20(d,J＝5.4Hz,1H,C ₆ -thienopyrimidine-H),8.00(s,1H,triazol-H ), 7.90 (d, J=8.7 Hz, 2H, C ₃ , C ₅ -Ph'-H), 7.82 (d, J=8.7 Hz, 2H, C ₂ , C ₆ -Ph'-H), 7.72 ( s,2H,C ₃ ,C ₅ -Ph”-H),7.28(s,1H,C ₇ -thienopyrimidine-H),6.88(s,1H,NH),5.38(s,2H,triazol-CH ₂ ) ,3.62–3.48(m,3H),3.18(s,3H),2.85–2.82(m,2H),2.11(s,6H),1.91–1.55(m,4H),1.43–1.17(m,2H) . ¹³ C NMR (100 MHz, DMSO-d ₆ ) δ165.7, 162.4, 153.4, 143.3, 140.3, 139.8, 135.6, 133.2, 128.8, 126.5, 124.5, 119.5, 117.1, 112.5, 109.8, 52.6, 52.3, 9 ,31.6,16.2.ESI-MS: m/z 672.3[M+1] ⁺ . C ₃₂ H ₃₃ N ₉ O ₄ S ₂ (671.21).

3,5-Dimethyl-4-((2-((1-((1-(2-morpholino-2-oxoethyl)-1H-1,2,3-triazole-4- base)methyl)piperidin-4-yl)amino)thieno[3,2-d]pyrimidin-4-yl)oxy)benzonitrile (A9)

White solid, yield 93%, melting point 228-230°C. ¹ H NMR (400MHz,DMSO-d ₆ )δ8.20(d,J＝5.4Hz, 1H,C ₆ -thienopyrimidine-H),7.84(s,1H,triazol-H),7.72(s,2H,C ₃ ,C ₅ -Ph”-H), 7.27(s,1H, C ₇ -thienopyrimidine-H), 6.87(s,1H,NH), 5.44(s,2H,triazol-CH ₂ ), 3.64(d, J＝4.8Hz, 2H), 3.59 –3.52(t, J＝4.9Hz, 3H), 3.53(d, J＝5.2Hz, 4H), 3.46(d, J＝4.8Hz, 2H), 2.79(s, 2H),2.11(s, 6H),1.90–1.51(m,4H),1.47–1.23(m,2H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ165.0,162.3, 153.7,143.3,140.3,139.8 ,135.6,133.2,125.8,117.5,112.1,109.6,66.4,66.3,53.1,52.3,50.9, 45.1,42.3,31.6,16.2.ESI-MS: m/z 588.5[M+1] ⁺ .C ₂₉ H _{33 N9O3S} ( _587.24 ).

3,5-Dimethyl-4-((2-((1-((1-(2-oxo-2-(pyrrolidin-1-yl)ethyl)-1H-1,2,3- Triazol-4-yl)methyl)piperidin-4-yl)amino)thieno[3,2-d]pyrimidin-4-yl)methyl)benzonitrile (A10)

White solid, yield 91%, melting point 219-221°C. ¹ H NMR (400MHz,DMSO-d ₆ )δ8.20(d,J＝5.4Hz, 1H,C ₆ -thienopyrimidine-H),7.84(s,1H,triazol-H),7.72(s,2H,C ₃ ,C ₅ -Ph”-H),7.27(s,1H,C ₇ -thienopyrimidine-H),6.87(s,1H,NH),5.31(s,2H,triazol-CH ₂ ),3.69(s, 1H), 3.52(t, J=6.8 Hz, 4H), 3.51–3.47(s, 2H, N-CH ₂ ), 3.40–3.24(m, 4H), 2.79(s, 2H), 2.11(s, 6H ),2.02–1.62(m, 4H),1.53–1.39(s,2H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ164.2,162.1,153.0,143.6,140.1,139.5,135.6,133.2,133.0,125.7 ,119.0,53.0,52.3,51.6,46.2,45.5,39.6,31.6,26.0,24.1,16.2. ESI-MS: m/z 572.5[M+1] ⁺ .C ₂₉ H ₃₃ N ₉ O ₂ S(571.25) .

Embodiment 5: In vitro anti-HIV activity test experiment of the target compound

Test principle:

The anti-HIV activity of compounds was screened in vitro by MTT method. The full name of MTT is bromide-3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium nitrogen, which can be used to detect the survival and growth of cells. The detection principle is: MTT can combine with living intracellular succinate dehydrogenase to reduce to water-insoluble blue-purple crystalline formazan and deposit in cells, while dead cells do not have this function. Dimethyl sulfoxide can dissolve formazan in the cells, and its absorbance (A) value at 540nm can indirectly reflect the number of living cells with a microplate reader. Within a certain cell number range, the amount of MTT crystal formation is proportional to the cell number.

Because the MT-4 cell that HIV infects can take place lesion within a certain period of time (5-7 days), therefore add the compound solution to be detected of appropriate concentration in the MT-4 cell suspension liquid that HIV infects, after a period of time (5-7 days) After 7 days) of culture, the viability of MT-4 cells was measured by MTT analysis method to obtain the drug concentration EC ₅₀ that protects 50% of the cells from cytopathic changes, and at the same time obtain the concentration CC of the target compound that causes 50% of HIV-uninfected cells to undergo pathological changes ₅₀ , to calculate the selection coefficient SI (SI=CC ₅₀ /EC ₅₀ ).

Test materials and methods:

(1) HIV-1 (III _B ), HIV-2 (ROD) strains, and HIV-1 double mutant strain RES056: provided by the Rega Institute, Faculty of Medicine, University of Leuven, Belgium.

(2) MT-4 cells: provided by the Rega Research Institute, Faculty of Medicine, University of Leuven, Belgium.

(3) MTT: purchased from Sigma, USA.

(4) Sample treatment: Before use, the sample was dissolved in DMSO to make an appropriate concentration, and diluted 5 times with double distilled water, each with 5 dilutions.

(5) Positive control drugs: nevirapine (NVP) and etravirine (ETV).

(6) Test method: the sample is diluted and added to the HIV-infected MT-4 cell suspension, and after a period of time, the cell viability is measured by the MTT colorimetric method, and the absorbance (A) value at 540nm is recorded in a microplate reader , calculate EC ₅₀ , CC ₅₀ and SI.

(7) MTT colorimetric method: After adding the sample solution and incubating for a period of time, add 20 μL of MTT solution (5 mg/mL) to each well, continue to cultivate for several hours, discard the staining solution, and add 150 μL DMSO to each well, and use enzyme Measure the absorbance (A) value at 540nm in the standard instrument.

experimental method:

Add 50 μL of 1×10 ⁴ MT-4 cell culture medium into a 96-well cell culture plate, add 20 μL of MT-4 cell mixture infected with HIV-1 (III _B , single mutant or double mutant) or HIV-2 (ROD) Suspension or blank culture medium, add different concentrations of test compound or positive control drug solution, and set 3 replicate wells for each concentration. The cells were cultured at 37°C for 5 days in a 5% _CO2 atmosphere, and 20 mL (5 mg/mL) MTT solution was added to each well, and the culture was continued for 2 hours, then DMSO was added, and the absorbance of the reaction solution at 540 nm was measured, and the difference between the compounds was calculated. The cell proliferation rate P% under the concentration. At the same time, a blank and a positive drug control group were set up, and the concentration required for the compound to protect 50% of the cells from HIV-induced cytopathy (EC ₅₀ ) and the concentration of the compound to cause 50% of HIV-uninfected cells to undergo pathological changes (CC ₅₀ ) were calculated. Calculation of selection index: SI=CC ₅₀ /EC ₅₀ .

The anti-HIV-1 (III _B ) and double mutant strain RES056 (K103N/Y181C) activity screening at the cell level was carried out on the synthesized part of the diarylpyrimidine derivatives containing triazole rings according to the above-mentioned experimental method, and the activity results As shown in Table 1.

Table 1. Anti-HIV activity, toxicity and selection index of some diarylpyrimidines containing triazole rings

^a EC ₅₀ : the compound concentration that inhibits 50% of virus-induced mutagenic effects or protects 50% of virus-infected cells from cytopathic effects.

^b CC ₅₀ : the concentration that makes 50% of HIV-uninfected cells pathological.

^c SI: Selection coefficient, ratio of _CC50 / _EC50 .

Claims (5)

1. A diarylpyrimidine HIV-1 inhibitor containing a triazole ring, or a pharmaceutically acceptable salt thereof, characterized in that it has a structure shown in general formula I: in, X is: O or MH; R is: CH ₃ , CN or CH=CHCN; R ¹ is: phenyl or pyridyl; or halogen, SO ₂ NH ₂ , SO ₂ CH ₃ , CONH ₂ , NO ₂ , CN, NH ₂ , CF ₃ , NHCH ₃ , OH, COOH, CH ₂ OH, CO ₂ Me, OCH ₃ , NHCOCH ₃ substituted phenyl; the substituent is ortho-, meta-, para-position monosubstituted or multi-substituted; or CONR ² ; Among them, R ² is: phenyl; or halogen, SO ₂ NH ₂ , SO ₂ CH ₃ , CONH ₂ , NO ₂ , CN, NH ₂ , CF ₃ , NHCH ₃ , OH, COOH, CH ₂ OH, CO ₂ Me , OCH ₃ , NHCOCH ₃ substituted phenyl; the substituents are ortho, meta, para mono-substituted or multi-substituted; five-membered heterocycle, six-membered heterocycle. 2. The diaryl pyrimidine HIV-1 inhibitor containing triazole ring as claimed in claim 1, characterized in that, it is one of the following compounds: 3. the preparation method of the diaryl pyrimidine class HIV-1 inhibitor containing triazole ring as claimed in claim 1, the step comprises: thieno[3,2-d] substituted with 2,4-dichloro Pyrimidine 1 is the initial raw material. First, it reacts with substituted phenol or aniline in N,N-dimethylformamide solution to generate intermediate 2 through nucleophilic substitution reaction; then intermediate 2 reacts with N-Boc-4-aminopiperidine through hydrophilic The nuclear substitution reaction generates intermediate 3, and then removes the Boc protection in trifluoroacetic acid to obtain intermediate 4; then reacts 4 with propyne bromide to generate the key intermediate 5, and then undergoes monovalent copper-catalyzed azide with the azide substituent - terminal alkyne cycloaddition reaction (CuAACClick) reaction obtains target product formula I compound; Synthetic route is as follows: Reagents and conditions: (i) substituted phenol or aniline, N,N-dimethylformamide, potassium carbonate, room temperature; (ii) N-Boc-4-aminopiperidine, N,N-dimethylformamide, Potassium carbonate, 110°C; (iii) dichloromethane, trifluoroacetic acid, room temperature; (iv) propyne bromide, N,N-dimethylformamide, potassium carbonate, room temperature; (v) sodium vitamin C, pentahydrate Copper sulfate, N,N-dimethylformamide, water, azide substituent, 50°C; X, R, R ¹ are shown in general formula I in claim 1; The substituted phenols or anilines are: s-trimethylphenol, 2,6-dimethyl-4-cyanophenol, 2,6-dimethyl-4-(E)-cyanovinylphenol, Trimethylaniline, 2,6-dimethyl-4-cyanoaniline, 2,6-dimethyl-4-(E)-cyanovinylaniline; The azide substituents are: o-chlorobenzyl azide, m-chlorobenzyl azide, p-chlorobenzyl azide, o-bromobenzyl azide, m-bromobenzyl azide, p-bromobenzyl azide , o-fluorobenzyl azide, m-fluorobenzyl azide, p-fluorobenzyl azide, 2,4-difluorobenzyl azide, 3,4-difluorobenzyl azide, o-cyanobenzyl azide Nitrogen, m-cyanobenzyl azide, p-cyanobenzyl azide, o-nitrobenzyl azide, m-nitrobenzyl azide, p-nitrobenzyl azide, o-methoxybenzyl azide , m-methoxybenzyl azide, p-methoxybenzyl azide, p-methylsulfonyl benzyl azide, m-methylsulfonyl benzyl azide, o-methylsulfonyl benzyl azide, p-sulfonyl benzyl azide Benzyl azide, m-sulfonamidobenzyl azide, o-sulfonamidobenzyl azide, p-carboxamidobenzyl azide, m-carboxamidobenzyl azide, o-carboxamidobenzyl benzyl azide , 2-azido-1-(pyrrolidin-1-yl)ethan-1-one, 2-azido-N-cyclopropylacetamide, 2-azido-1-morpholine-1- ketone. 4. Use of the diarylpyrimidine HIV-1 inhibitor containing a triazole ring as claimed in claim 1 or 2 in the preparation of anti-AIDS drugs. 5. An anti-AIDS pharmaceutical composition, comprising the diaryl pyrimidine HIV-1 inhibitor containing a triazole ring described in claim 1 or 2 and one or more pharmaceutically acceptable carriers or excipients .