S-daco non-nucleoside reverse transcriptase inhibitor derivatives and uses thereof

Abstract

The application discloses a kind of S-DACOs non-nucleoside reverse transcriptase inhibitor derivatives and its pharmaceutically acceptable salt, structural formula is as follows: wherein, R a selected from NH2, SO2Me, wherein R1 is selected from H, C1-C6 linear or branched alkyl, alkenyl, R2 is selected from H, C1-C6 linear or branched alkyl, alkenyl;R b selected from H, C1-C3 linear or branched alkyl;The application carries out structural modification to the C-2 terminal benzene ring of S-DACOs, introduces suitable substituent, improves the lipophilic water partition coefficient of drug while retaining high efficient anti-HIV activity, improves its drugability, and the compound of the application can be used for preparing medicine for treating and / or preventing immunodeficiency virus.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, and particularly relates to derivatives of S-DACOs class of non-nucleoside reverse transcriptase inhibitors (NNRTIs) and application thereof, which can be used as HIV-1 inhibitors and for preparing a drug for treating and / or preventing immunodeficiency virus (HIV). BACKGROUND

[0002] Acquired immunodeficiency syndrome (AIDS) is a disease caused by human immunodeficiency virus (HIV) in the human body, which causes the immune system to gradually weaken, and is simply referred to as AIDS. Since the first case of HIV was reported in 1981, HIV has spread worldwide, and the number of deaths has exceeded 20 million. Although the cocktail therapy can effectively inhibit the replication of HIV virus and reduce the mortality, it still cannot completely eliminate the HIV virus, so it is an urgent problem to be solved in the treatment of AIDS to develop an anti-HIV drug with high efficiency, low toxicity and low price. Reverse transcription catalyzed by reverse transcriptase (RT) is a key stage in the life cycle of HIV, during which genetic information is transmitted from a single strand of viral RNA to a double strand of host cell DNA, and has always been the focus of pharmacists in the research of anti-HIV drugs. Reverse transcriptase inhibitors are divided into nucleoside (NRTls) and non-nucleoside (NNRTls), and NNRTls have always been the focus of anti-HIV drug research due to their low toxicity and high efficiency.

[0003] S-DACOs class of NNRTIs is a kind of anti-HIV active compounds designed and synthesized by the research group using computer-aided drug molecule design method. The compounds can inhibit HIV replication in a lower nanomolar range, have lower cytotoxicity, and have excellent selectivity index. However, the S-DACOs class of compounds has poor water solubility due to the large number of hydrophobic groups in the structure, resulting in low bioavailability, which affects its drugability. DB02 is the best anti-HIV active molecule in the S-DACOs class of compounds, which has high inhibitory activity on a variety of HIV-1 experimental strains and mutant strains, but has poor water solubility (S = 0.38 μM), and the pharmacokinetic properties are not ideal, which limits its further development and application.

[0004]

[0005] It is found through molecular docking research that the binding force between the S-DACO class of inhibitors represented by DB02 and the RT enzyme is mainly (1) hydrogen bond interaction between β-carbonyl at C-2, 3-NH and surrounding amino acid residues; (2) hydrophobic interaction and van der Waals force between C-6 cyclohexyl and the end of C-2 side chain aromatic ring and surrounding amino acid residues.Figure 1 Meanwhile, we noticed that the C-2 side chain of DB02 is located in a flexible hydrophobic pocket composed of P236 and V106, and the terminal benzene ring is close to the opening of the RT enzyme active pocket, with a large space around to accommodate a structurally diverse substituent, which is an important structural modification site for adjusting the physicochemical and pharmacokinetic properties of active molecules.

[0006] A novel S-DACOs class of non-nucleoside reverse transcriptase inhibitors (NNRTIs) is disclosed in ZL 200710066433.7, which has the following structure: In the structure, R1 is C 1-3 alkyl, X = OCH3, H, OH, halogen. This compound is a highly efficient and low-toxic HIV-1 reverse transcriptase inhibitor, but has poor water solubility, leading to low oral bioavailability, which affects its drugability. SUMMARY

[0007] The present application provides a class of S-DACOs class of non-nucleoside reverse transcriptase inhibitors (NNRTIs) derivatives with good water solubility, better lipid-water partition coefficient, and strong anti-HIV activity, which has the structure as shown in formula I:

[0008]

[0009] wherein R a is selected from NH2, SO2Me, wherein R1 is selected from H, C1-C6 linear or branched alkyl, alkenyl, R2 is selected from H, C1-C6 linear or branched alkyl, alkenyl; R b is selected from H, C1-C3 linear or branched alkyl.

[0010] The present application is based on molecular docking analysis, and the C-2 terminal benzene ring of S-DACOs is structurally modified by introducing a suitable substituent, while retaining high anti-HIV activity, improving the lipid-water partition coefficient of the drug, and improving its drugability.

[0011] The S-DACOs class of non-nucleoside reverse transcriptase inhibitor derivatives of the present application can be used as HIV-1 inhibitors and for the preparation of drugs for treating and / or preventing immunodeficiency virus (HIV) infection.

[0012] The S-DACOs class of non-nucleoside reverse transcriptase inhibitor derivatives of the present application also includes pharmaceutically acceptable salts of the S-DACOs class of non-nucleoside reverse transcriptase inhibitor derivatives.

[0013] The S-DACOs class of NNRTIs derivatives of the present application are selected from any one of the following compounds:

[0014]

[0015]

[0016] The preparation method of the phosphate derivative in the S-DACOs class NNRTIs derivative (when R a is The preparation method of the phosphate derivative in the S-DACOs class NNRTIs derivative (when R

[0017]

[0018] The molar ratio of the compound 3 to the phosphoryl chloride compound 5 is 1:1-4; the basic reagent is one or more of triethylamine, K2CO3, NaHCO3, NaH, NaOCH3, NaOEt and Et3N; and the solvent is one or more of tetrahydrofuran, toluene, acetonitrile, dichloromethane, dimethylformamide and pyridine.

[0019] The reaction process can be monitored by using conventional test methods in the art (such as TLC, HPLC or NMR), and the reaction is generally terminated when the raw material disappears or no longer reacts, and the reaction time is 1-4 hours;

[0020] In the above reaction, the compound 3 is prepared by reacting the compound 1 with p-hydroxy bromoacetophenone in the presence of a solvent and a weak basic reagent, and the reaction formula is as follows:

[0021]

[0022] The compound 1 is dissolved in DMF, a weak basic reagent K2CO3 is added, and then a mixed solution of p-hydroxy bromoacetophenone and DMF is added after stirring, and the reaction is continued after stirring. After the reaction is completed, the reaction liquid is poured into ice water, and white turbidity is generated after vigorous stirring. The filter cake is washed with ice water for 3 times, and is dried to obtain the crude product of the compound 3, which can be directly used in the next step reaction without purification.

[0023] In the present application, the compound 1 can be prepared by using the method well known to those skilled in the art of organic chemistry, and in the present application, reference can be made to Yan-Ping He, Jin Long, et al. Bioorg. & Med. Chem. 2011, 21, 694-697, wherein the third paragraph on page 695 and Zhi-Kun Rao, Jing Long, et al. Monatsh Chem. 2008, 139, 967-974, and the specific synthesis route is as shown below:

[0024]

[0025] In the present application, the phosphorus oxychloride compound 5 is prepared by dissolving dialkyl phosphite 4 with CCl4, slowly adding TEA, stirring at room temperature for 30 minutes, filtering the reaction solution, washing the filter cake with acetonitrile for 3 times, and distilling the filtrate under reduced pressure to obtain compound 5.

[0026]

[0027] wherein R1, R2, R b have the same meanings as defined above.

[0028] According to the above preparation method disclosed in the present application, those skilled in the art can use the same principles and methods to prepare each specific compound involved in the compound of formula I of the present application.

[0029] In the present application, the compound of formula I can be prepared by the following method when R a is NH2 or SO2Me.

[0030]

[0031] Advantages and technical effects of the present application:

[0032] 1. The compound preparation method of the present application is simple and can realize industrial production.

[0033] 2. The compound of the present application has significant anti-HIV activity through in vitro cell level anti-HIV activity experiment, and its inhibitory activity on HIV III is better than that of the lead compound DB02 and the positive control 3TC.

[0034] 3. The water solubility of the S-DACOs derivative of the present application is significantly improved, and the results of molinspiration software calculation show that it has better lipid-water partition coefficient and pharmacokinetic properties than the existing S-DACOs class NNRTIs. BRIEF DESCRIPTION OF DRAWINGS

[0035] Figure 1 is a DB02 / HIV RT complex model. DETAILED DESCRIPTION

[0036] The present application will be described in detail below by way of examples, but it does not mean any unfavorable limitation of the present application. The present application has been described in detail herein, and the specific embodiment modes thereof have also been disclosed, and it will be obvious to those skilled in the art to make various changes and improvements to the specific embodiment modes of the present application without departing from the spirit and scope of the present application.

[0037] The experimental methods in the following examples, unless otherwise specified, are selected according to the conventional methods and conditions, or according to the commercial instruction. The raw materials can be obtained from commercial channels, or prepared by the methods known in the art, or prepared according to the methods described herein. The structure of the compound is determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS), wherein the NMR determination uses a Bruker DRX 400 type nuclear magnetic resonance instrument, the determination solvent is deuterated pyridine (C5D5N), and TMS is an internal standard. 1 H NMR or 13 C NMR) and mass spectrometry (MS), wherein the NMR determination uses a Bruker DRX 400 type nuclear magnetic resonance instrument, the determination solvent is deuterated pyridine (C5D5N), and TMS is an internal standard.

[0038] Example 1: Preparation of target compounds I-1 to I-8

[0039]

[0040] In a 100 mL dry two-necked round-bottom flask, 10 mmol of phosphorus oxychloride 5 and 6 mmol of TEA were added, then dissolved with 50 mL of DCM, 10 mmol of compound 3 was added under ice bath, after the addition was completed, the ice bath was removed, and the reaction was carried out at room temperature for 2 hours. TLC tracking showed that the reaction was complete. 1 mol / L HCl solution was added to quench the reaction. The lower organic phase was removed, and the upper solution was extracted with 50 mL of DCM for 3-4 times. The organic phases were combined, neutralized with 50 mL of saturated sodium bicarbonate solution, washed with 50 mL of saturated brine for 3 times, dried over anhydrous sodium sulfate, and distilled under reduced pressure to obtain the crude product. Then, the target compound I-1 was obtained as a white powder solid by silica gel column chromatography (eluent ethyl acetate: petroleum ether = 5:1), and the yield was 38%.

[0041]

[0042] 1 H NMR (400 MHz, Pyridine-d5) δ (ppm): 10.41 (s, 1H, -NH), 7.93 (d, J = 8.5 Hz, 2H, Ph-H), 6.87 (d, J = 8.5 Hz, 2H, Ph-H), 4.58 (s, 2H, CH2-S), 2.29 (q, J = 7.3 Hz, 2H, CH2), 2.10 (d, J = 6.3 Hz, 2H, CH2-Cyclohexyl), 1.47 (d, J = 8.6 Hz, 3H, Cyclohexyl), 1.30 (d, J = 13.2 Hz, 3H, Cyclohexyl), 0.99-0.84 (m, 6H, overlap), 0.71 (q, J = 11.5 Hz, 2H, Cyclohexyl); HR-MS: m / z calcd for C 21 H 27 N2O6PS[M+H] +: 467.1405; found 467.1401.

[0043] The preparation method of the compounds of formula I-2 to I-8 is only different from the preparation of the compound of formula I-1 in that the intermediate 3 or the phosphorus oxychloride compound 5 is different, wherein the intermediate 3 is The phosphorus oxychloride compound 5 is In addition, the rest of the preparation method is the same as the preparation method of the compound of formula I-1 in this example.

[0044] The properties, yield and structural characterization results of the prepared compounds of formula I-2 to I-8 are as follows:

[0045]

[0046] The operation is as above, and white powder solid I-2 is obtained, the yield is 55%;

[0047] 1 H NMR (400 MHz, Pyridine-d5) δ (ppm): 8.13 (d, J = 8.7 Hz, 2H, Ph-H), 7.39 (d, J = 8.2 Hz, 2H, Ph-H), 4.65 (s, 2H, CH2-S), 3.84 (s, 3H, CH3-O), 3.81 (s, 3H, CH3-O), 2.29 (q, J = 7.2 Hz, 2H, CH2), 2.07 (d, J = 6.4 Hz, 2H, CH2-Cyclohexyl), 1.47-1.44 (m, 3H, Cyclohexyl), 1.26-1.23 (m, 4H, Cyclohexyl), 0.92 (t, J = 7.3 Hz, 3H, Et), 0.87-0.61 (m, 4H, Cyclohexyl); HR-MS: m / z calcd for C 23 H 31 N2O6PS[M+H] + : 495.1713; found 495.1707.

[0048]

[0049] The operation is as above, and white powder solid I-3 is obtained, the yield is 55%;

[0050] 1H NMR (400 MHz, Pyridine-d5) δ (ppm): 8.13 (d, J = 8.6 Hz, 2H, Ph-H), 7.38 (d, J = 8.5 Hz, 2H, Ph-H), 4.65 (s, 2H, CH2-S), 4.19 (q, J = 8.0, 7.6 Hz, 4H, CH2), 2.29 (q, J = 7.1 Hz, 2H, CH2), 2.07 (d, J = 6.2 Hz, 2H, CH2-Cyclohexyl), 1.46 - 1.44 (m, 3H, Cyclohexyl), 1.29 - 1.23 (m, 11H, overlap), 0.93 - 0.85 (m, 4H, Cyclohexyl), 0.71 - 0.67 (m, 2H, Cyclohexyl); HR-MS: m / z calcd for C 25 H 35 N2O6PS[M+H] + :523.2026; found 523.2031.

[0051]

[0052] Following the procedure as above, white powder solid I-4 was obtained in 41% yield;

[0053] 1 H NMR (400 MHz, Pyridine-d5) δ (ppm): 8.13 (d, J = 8.5 Hz, 2H, Ph-H), 7.38 (d, J = 8.5 Hz, 2H, Ph-H), 4.65 (s, 2H, CH2-S), 4.19 (q, J = 8.0, 7.6 Hz, 4H, CH2), 2.29 (q, J = 7.1 Hz, 2H, CH2), 2.07 (d, J = 6.2 Hz, 2H, CH2-Cyclohexyl), 1.46 - 1.44 (m, 3H, Cyclohexyl), 1.29 - 1.23 (m, 11H, overlap), 0.93 - 0.85 (m, 4H, Cyclohexyl), 0.71 - 0.67 (m, 2H, Cyclohexyl); HR-MS: m / z calcd for C 29 H 43 N2O6PS[M+H] + :579.2632; found 579.2635.

[0054]

[0055] Operation as above, white powder solid I-5 was obtained, yield: 53%;

[0056] 1 H NMR (400 MHz, Pyridine-d5) δ (ppm): 8.13 (d, J = 8.7 Hz, 2H, Ph-H), 7.38 (d, J = 8.3 Hz, 2H, Ph-H), 4.64 (s, 2H, CH2-S), 2.29 (s, 3H, CH3), 2.08 (d, J = 6.3 Hz, 2H, CH2-Cyclohexyl), 1.52 - 1.43 (m, 3H, Cyclohexyl), 1.35 - 1.21 (m, 8H, Cyclohexyl); HR-MS: m / z calcd for C 20 H 25 N2O6PS[M+H] + : 453.1235; found 453.1237.

[0057]

[0058] Operation as above, white powder solid I-6 was obtained, yield: 58%;

[0059] 1 H NMR (400 MHz, Pyridine-d5) δ (ppm): 8.13 (d, J = 8.7 Hz, 2H, Ph-H), 7.38 (d, J = 8.3 Hz, 2H, Ph-H), 4.64 (s, 2H, CH2-S), 3.59 (s, 6H, O-CH3), 2.28 (s, 3H, CH3), 2.08 (d, J = 6.3 Hz, 2H, CH2-Cyclohexyl), 1.46 - 1.44 (m, 3H, Cyclohexyl), 1.30 - 1.13 (m, 5H, Cyclohexyl), 0.71 - 0.63 (m, 3H, Cyclohexyl); HR-MS: m / z calcd for C 22 H 29 N2O6PS[M+H] + : 481.1562; found 481.1563.

[0060]

[0061] Operation as above, white powder solid I-7 was obtained, yield: 50%

[0062] 1H NMR (400 MHz, Pyridine-d5) d (ppm): 7.92 (d, J = 8.5 Hz, 2H, Ph-H), 6.87 (d, J = 8.6 Hz, 2H, Ph-H), 4.59 (s, 2H, CH2-S), 2.33 (m, 1H, CH), 2.11 (d, J = 6.3 Hz, 2H, CH2-Cyclohexyl), 1.47 (d, J = 8.6 Hz, 3H, Cyclohexyl), 1.30 (d, J = 13.2 Hz, 3H, Cyclohexyl), 0.97 - 0.79 (m, 9H, overlap), 0.71 (q, J = 11.5 Hz, 2H, Cyclohexyl); HR-MS: m / z calcd for C 22 H 29 N2O6PS[M+H] + : 481.1518; found 481.1515.

[0063]

[0064] Following the procedure as above, white powder solid I-8 was obtained in 47% yield;

[0065] 1 H NMR (400 MHz, Pyridine-d5) d (ppm): 8.12 (d, J = 8.6 Hz, 2H, Ph-H), 7.39 (d, J = 8.3 Hz, 2H, Ph-H), 4.66 (s, 2H, CH2-S), 3.83 (s, 3H, CH3-O), 3.82 (s, 3H, CH3-O), 2.29 (m, 1H, CH), 2.07 (d, J = 6.4 Hz, 2H, CH2-Cyclohexyl), 1.47 - 1.44 (m, 3H, Cyclohexyl), 1.27 - 1.23 (m, 4H, Cyclohexyl), 0.92 (m, 6H, CH3), 0.87 - 0.61 (m, 4H, Cyclohexyl); HR-MS: m / z calcd for C 24 H 33 N2O6PS[M+H] + : 509.1801; found 509.1803.

[0066] Example 2: Preparation of target compound I-9

[0067] Take 5-ethyl-6 cyclohexylmethyl-2-thiouracil (0.01 mol) in a 25 mL round-bottom flask, dissolve with 10 mL of dry DMF, add 0.012 mol of anhydrous potassium carbonate, stir for 1 h, then add 0.01 mol of p-toluenesulfonyl bromoacetophenone, stop the reaction after tracking the disappearance of the raw material point by TCL, pour the reaction liquid into ice water (50 mL) and extract with ethyl acetate three times (25 mL each time), wash the combined organic phase with saturated brine three times (25 mL each time), dry over anhydrous sodium sulfate, rotary evaporation under reduced pressure, and then column chromatography to obtain white powder solid I-9, yield: 86%;

[0068]

[0069] 1 H NMR (500 MHz, Pyridine-d5, ppm) δ: 8.50 (2H, s, aromatic), 8.39 (2H, s, aromatic), 5.00 (2H, s, CH2-S), 3.42 (3H, s, CH3-Ph), 2.64 (2H, s, CH2-CH3), 2.27 (2H, s, CH2-CH), 2.00-1.96 (1H, m, CH), 1.61-1.54 (5H, m, cyclohexyl), 1.19-1.02 (6H, m, overlap), 0.83-0.78 (2H, m, cyclohexyl). HRMS-ESI: m / z calcd for C 22 H 29 N2O4S2[M+H] + : 449.1563, found 449.1563.

[0070] Example 3: Preparation of target compounds I-10 to I-13

[0071]

[0072] Take 0.01 mol of compound I-A in a 2 mL round-bottom flask and dissolve with 20 mL of ethanol, then add 0.05 mol of iron powder, dissolve 0.1 mol of ammonium chloride in 5 mL of pure water and add dropwise to the reaction liquid, stop the reaction after refluxing at 85°C for 2 h, filter the iron powder and wash with ethanol three times, pour the filtrate into ice water (50 mL) and extract with ethyl acetate three times (25 mL each time), wash the combined organic phase with saturated brine three times (25 mL each time), dry over anhydrous sodium sulfate, rotary evaporation under reduced pressure, and then column chromatography or recrystallization to obtain compounds I-10 to I-13;

[0073]

[0074] Pale yellow solid, yield 68%. 1 H NMR (400MHz, Pyridine-d5, ppm) δ: 8.24-8.22 (2H, d, J = 8.8Hz, aromatic), 7.01-6.99 (2H ,d,J=8.4Hz,aromatic),6.88(2H,s,NH2),4.96(2H,s,CH2-S),2.69-2.64(2H,q,J=7.2Hz ,CH2-CH3),2.42-2.41(2H,d,J=7.2Hz,CH2-CH),1.89-1.85(1H,m,CH),1.67-1.54(5H,m, cyclohexyl),1.24-1.10(6H,m,overlap),0.96-0.88(2H,m,cyclohexyl).HRMS-ESI:m / z calcd for C 21 H 28 N3O2S[M+H] + :386.1897,found386.1895.

[0075]

[0076] Yellow solid, yield 60%. 1 H NMR(400MHz,Pyridine-d5,ppm)δ:7.85(1H,s,aromatic),7.64-7.62(2H,d,J=8.0Hz,aromatic) ,7.38-7.34(2H,t,J=7.80Hz,aromatic),6.88(2H,s,NH2),4.95(2H,s,CH2-S),2.67-2.62(2H,t, J=7.3Hz,CH2-CH3),2.38-2.36(2H,q,J=6.8Hz,CH2-CH3),1.84-1.75(1H,m,CH),1.62-1.56(5H, m,cyclohexyl),1.26-1.22(6H,m,overlap),0.93-0.80(2H,m,cyclohexyl).HRMS-ESI:m / zcalcd for C 21 H 28 N3O2S[M+H] + 386.1897, found 386.1898.

[0077]

[0078] Yellow solid, yield 56%.1 H NMR (400 MHz, Pyridine-d5, ppm) δ: 8.16-8.14 (2H, d, J = 8.0 Hz, aromatic), 8.16-8.14 (2H, s, NH2), 7.38-7.34 (2H, t, J = 7.6 Hz, aromatic), 7.07-7.05 (2H, d, J = 8.8 Hz, aromatic), 6.74-6.70 (2H, t, J = 8.0 Hz, aromatic), 5.00 (2H, s, CH2-S), 2.70-2.64 (2H, q, J = 7.3 Hz, CH2-CH3), 2.43-2.41 (2H, d, J = 7.2 Hz, CH2cyclohexyl), 1.88-1.82 (1H, m, CH), 1.66-1.55 (5H, m, cyclohexyl), 1.24-1.08 (6H, m, overlap), 0.96-0.87 (2H, m, cyclohexyl). HRMS-ESI: m / z calcd for C 21 H 28 N3O2S [M+H] + 386.1897, found 386.1897.

[0079]

[0080] Yellow solid, yield 60%. 1 H NMR (400 MHz, Pyridine-d5, ppm) δ: 7.85 (1H, s, aromatic), 7.65-7.63 (2H, d, J = 8.0 Hz, aromatic), 7.38-7.35 (2H, t, J = 7.80 Hz, aromatic), 6.88 (2H, s, NH2), 4.95 (2H, s, CH2-S), 3.26-2.91 (1H, m, CHMe2), 2.43-2.41 (2H, d, J = 7.2 Hz, CH2cyclohexyl), 1.84-1.75 (1H, m, CH), 1.62-1.56 (5H, m, cyclohexyl), 1.26-1.29 (9H, m, overlap), 0.91-0.78 (2H, m, cyclohexyl). HRMS-ESI: m / z calcd for C 22 H 29 N3O2S [M+H] + 400.5534, found 400.5528.

[0081] Example 4: Preparation of target compound I-14

[0082] Take 50 mL round bottom flask, add 1 mmol compound I-11, add 5 mL CH2Cl2stir until completely dissolved, saturated amount of HC1 gas, constant temperature stirring, solid precipitated, TLC detection of raw material point disappeared, stop the reaction, filter, recrystallized with ethanol to get I-14 pure product.

[0083]

[0084] Light yellow solid, yield 95%. 1 H NMR (400 MHz, Pyridine-d5, ppm) δ: 7.83 (1H, s, aromatic), 7.62-7.60 (2H, d, J = 8.0 Hz, aromatic), 7.37-7.32 (2H, t, J = 7.80 Hz, aromatic), 6.87 (2H, s, NH2), 6.58 (bra, H Cl), 4.92 (2H, s, CH2-S), 2.65-2.61 (2H, t, J = 7.3 Hz, CH2-CH3), 2.36-2.34 (2H, q, J = 6.8 Hz, CH2-CH3), 1.82-1.73 (1H, m, CH), 1.61-1.56 (5H, m, cyclohexyl), 1.24-1.20 (6H, m, overlap), 0.92-0.78 (2H, m, cyclohexyl). HRMS-ESI: m / z calcd for C 21 H 29 ClN3O2S[M+H] + 422.6477, found 422.6474.

[0085] Example 5: Anti-HIV-1 activity test

[0086] In vitro cell level anti-HIV-1 activity test: the toxicity of the above prepared compound on C8166 cells was determined by MTT colorimetric method, the survival rate was calculated, and the concentration when 50% of the cells were toxic (50% cytotoxic concentration, CC 50 ). The protective effect of the compound on HIV-induced cytopathic effect was determined, the inhibition rate was calculated, and the concentration when 50% of the syncytia formation was inhibited (50% effective concentration, EC 50 ) was calculated. Then, the therapeutic index TI value (CC 50 / EC 50 ) was calculated, and the specific method was as follows:

[0087] Cytotoxicity experiment: the test compound was diluted 5 times, a total of 6 gradients, 100 μL of compound was added to each well of a 96-well plate, and triplicate wells were set up, then 4 x 10 4 6cells were added to each well, 3TC was used as a positive control drug, and a negative control containing only cells and a blank control containing only medium were also set up; after incubation at 37°C in a 5% CO2 incubator for 3 days, 20 μL of MTT solution (5 μg / mL) was added to each well and incubated for 4 h, 100 μL of supernatant was discarded from each well, and 12% SDS-50% DMF solution was added, and incubated overnight; the OD value was determined using an ELx800 enzyme marker, the determination wavelength was 570 nm / 630 nm, and the cell survival rate and CC 50 value were calculated.

[0088] Syncytia formation inhibition experiment: the test compound was diluted 5 times, 100 μL was added to each well of a 96-well plate, then 100 μL of a suspension containing 4 x 10 4 6cells and HIV-1 ⅢB (MOI = 0.03) was added to each well, triplicate wells were set up, and a control well containing only the cell-virus suspension was set up, 3TC was used as a positive drug control; then incubation was carried out at 37°C in a 5% CO2 incubator for 3 days, syncytia were counted under an inverted microscope (100x), and the inhibition rate and EC 50 value were calculated.

[0089] The present application uses 3TC as a positive control drug, and the results of the inhibitory activity of the target compound on HIV-1 IIIB are shown in Table 2:

[0090] Table 2 Anti-HIV-1 ⅢB viral activity of compounds

[0091]

[0092]

[0093] As can be seen from Table 2, the phosphate ester derivatives of S-DACOs have excellent anti-HIV III activity, and the EC 50 values of the remaining compounds are lower than that of the lead compound DB02 (EC 50= 0.2 μM), wherein the activity of compounds I-2 and I-3 were 2.2 and 11 times higher than that of the positive control 3TC, respectively. The introduction of amino (-NH2) and methylsulfonyl (-SO2Me) at the end of the C-2 side chain phenyl ring of the DB02 pyrimidine ring also significantly improved the anti-HIV activity, and the activity of compounds I-9 to I-14 was 3.7 to 50 times higher than that of the lead compound DB02 and the positive control 3TC.

[0094] Example 6: Drug solubility detection and pharmacokinetic property evaluation

[0095] A buffer solution of pH = 7.4, 100 mM was prepared by adding sodium dihydrogen phosphate, disodium hydrogen phosphate and sodium hydroxide to pure water; 8 μL of standard (Ketoconazole) and 10 mM of test compound stock solution were added to 792 μL of buffer solution, and the sample tube was shaken at room temperature on a shaker at 1000 rpm for 1 h; a standard curve was prepared in a methanol: ACN (4:1 by volume) solution; the sample tube was centrifuged, and the supernatant was diluted 10 times and 100 times with the above buffer solution, respectively; 5 μL of sample and standard curve sample (undiluted, diluted 10 times, diluted 100 times) were added to 95 μL of acetonitrile (containing IS) to prepare HPLC samples, which were detected by HPLC to calculate the solubility of the compounds; the solubility of some of the compounds of the application measured by this method is shown in Table 3.

[0096] The "five-fold rule" or "Lipinski five-fold rule" is an empirical rule for evaluating whether a compound can be a drug, or a compound with pharmacological activity or biological activity can be an oral drug. The rule states that a compound with drug potential has a logarithmic value of the octanol-water partition coefficient (ClogP) of no more than 5; generally, if the ClogP value of a compound is between 2 and 4, it is predicted to have good pharmacokinetic properties; the CLogP of compounds I-1 to I-14 was calculated using the molinspiration software, and the results are shown in Table 3:

[0097] Table 3 Solubility of compounds and CLogP values

[0098]

[0099] As shown in Table 3, the water solubility of compounds I-1 to I-8 is increased by 443 to 7 times compared with that of DB02 after phosphorylation modification, except for I-4; the solubility of mesyl derivative I-9 is increased by 158 times compared with that of DB02. The solubility of amino derivatives I-10 to I-13 is only increased by about 16 times compared with that of DB02, and the hydrochloride salt I-14 prepared by using the alkalinity of amino has a solubility of 137.21 μmol / L, which is increased by 361 times compared with that of DB02. The calculation results by using molinspiration software show that the ClogP values of the 14 S-DACOs class NNRTIs derivatives shown in Table 3 are all decreased compared with that of DB02, especially the ClogP values of compounds I-1, I-5, I-6, I-7 and I-9 are in the range of 2.25 to 3.91, indicating that these compounds have excellent pharmacokinetic properties.

[0100] The applicant declares that the S-DACOs class NNRTIs derivative and use of the present application are illustrated by the above examples, but the present application is not limited to the above examples, i.e. it does not mean that the present application must rely on the above examples to be implemented. It should be understood by those skilled in the art that any improvement of the present application, equivalent replacement of each raw material of the product of the present application, addition of auxiliary ingredients, selection of specific modes, etc. all fall within the protection scope and disclosure scope of the present application.

Claims

1. S-DACOs class of non-nucleoside reverse transcriptase inhibitor derivatives of the structural formula as shown in Formula I and pharmaceutically acceptable salts thereof; ###0001### Formula I wherein R a selected from NH2, SO2Me, wherein R1is selected from H, C1-C6 linear or branched alkyl, R2is selected from H, C1-C6 linear or branched alkyl; R b selected from H, C1-C3 linear or branched alkyl.

2. The S-DACOs class of non-nucleoside reverse transcriptase inhibitor derivatives and pharmaceutically acceptable salts thereof according to claim 1, characterized in that, which is any one of the following compounds:

3. Use of the S-DACOs class of non-nucleoside reverse transcriptase inhibitor derivatives of claim 1 and pharmaceutically acceptable salts thereof in the manufacture of a medicament for the treatment and / or prophylaxis of human immunodeficiency virus infection.

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