Isopropylpyridine-derived 1, 3-hydroxyl / acetoxy compound as well as synthesis method and application thereof

Abstract

The invention discloses an isopropyl pyridine derivative 1, 3-hydroxyl / acetoxyl compound as well as a synthesis method and application thereof, and belongs to the field of organic chemistry. The preparation method comprises the following steps: reacting 2-[2-(2-fluorophenyl)-1, 1-dimethyl ethyl] pyridine / 2-[2-(2-chlorphenyl)-1, 1-dimethyl ethyl] pyridine as a raw material and dichloromethane as a reaction solvent in the presence of palladium acetate, iodobenzene diacetate, water and acetic acid under the protection of nitrogen to obtain the isopropyl pyridine derivative 1, 3-hydroxyl / acetoxy compound with antiviral activity. The method is simple in synthesis step and environmentally friendly, palladium acetate is used for catalyzing the reaction, water which is low in price, easy to obtain, safe and harmless is used as a hydroxyl source, and the derivative has an obvious inhibition effect on the Zika virus and is expected to be used in potential anti-Zika virus drugs.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of organic chemistry, and particularly relates to isopropyl pyridine derivative 1,3-hydroxy / acetoxy compounds and a synthesis method and application thereof. BACKGROUND

[0002] 1,3-difunctionalization is an important reaction strategy in organic chemistry, and plays an important role in the construction of complex organic molecular structure, drug synthesis, material chemistry and the like. 1,3-difunctionalization reaction can introduce two valuable functional groups at specific 1,3-positions in a molecule, thereby increasing the diversity and complexity of the molecule, and providing a simple and efficient tool for chemists to synthesize complex compounds. Chlorine atom, hydroxyl and acetoxy exist in most organic compounds, and are widely present in drug molecules and material molecules. Similarly, they play an important role in the field of organic synthesis, and are intermediates for synthesizing many complex molecules, and can be converted into various groups by using their excellent reactivity.

[0003] In recent years, transition metal-catalyzed carbon-hydrogen bond activation reaction provides many examples of installing chlorine atom, hydroxyl and acetoxy, but these examples can only install one kind of functional group or install two different functional groups step by step, and there is no report on one-pot synthesis of hydroxyl / acetoxy in a single system.

[0004] In view of the current research progress of 1,3-difunctionalization, the present application designs and researches a method for one-pot synthesis of hydroxyl / acetoxy 1,3-difunctional pyridine derivatives in a single catalytic system promoted by palladium-catalyzed high-valence iodine. The reaction has mild reaction conditions, simple and readily available raw materials, is green and environmentally friendly, and has a wide range of reaction substrates. On this basis, biological activity research is carried out, and it is found that it has anti-zika virus activity, and has broad application prospects in anti-zika virus. SUMMARY

[0005] In order to overcome the above technical defects, the present application provides a 1,3-hydroxy / acetoxy product of isopropyl pyridine derivative, and researches a synthesis method and activity application of the compound.

[0006] The isopropyl pyridine derivative 1,3-hydroxy / acetoxy compound described in the present application has the following structural formula: , In the formula, R is selected from m-fluorobenzyl or m-chlorobenzyl.

[0007] Further, the isopropyl pyridine derivative 1,3-hydroxy / acetoxy compound specifically has the following structure: .

[0008] The application also provides a synthesis method of the isopropyl pyridine derivative 1,3-hydroxy / acetoxy compound, comprising the following steps: adding TDL-S1 into an organic solvent, and reacting in the presence of palladium acetate, iodobenzene acetate, water and acetic acid to obtain the isopropyl pyridine derivative 1,3-hydroxy / acetoxy compound, and the reaction equation is as follows: , wherein R is selected from m-fluorobenzyl or m-chlorobenzyl.

[0009] Further, in the above technical solution, the organic solvent is selected from dichloromethane, chloroform or 1,2-dichloroethane.

[0010] Further, in the above technical solution, the reaction temperature is 40-60 ℃.

[0011] Further, in the above technical solution, the molar ratio of the compound TDL-S1, palladium acetate, iodobenzene acetate, water and acetic acid is 1:0.1:2:2:10.

[0012] The application also provides an application of the isopropyl pyridine derivative 1,3-hydroxy / acetoxy compound in preparation of an antiviral drug.

[0013] Further, in the above technical solution, the virus is Zika virus.

[0014] The application adopts one-pot synthesis, and the reaction raw material is easy to obtain, the synthesis steps are simple, and the reaction condition is mild. Under the condition of nitrogen protection, 2-[2-(2-fluorophenyl)-1,1-dimethylethyl]pyridine and 2-[2-(2-chlorophenyl)-1,1-dimethylethyl]pyridine are used as raw materials, palladium acetate is used as a catalyst, iodobenzene acetate is used as an oxidant, dichloromethane is used as a solvent, water and acetic acid are sequentially added, and the product is obtained by reacting in a 50 ℃ metal bath for 12 hours. In the reaction, water is cheap, easy to obtain, safe and harmless, and is used as a source of hydroxyl group. Acetic acid and iodobenzene acetate are green and cheap additives, which shows the advantages of green and environmental protection of the reaction. In addition, part of the product has an inhibitory effect on Zika virus. BRIEF DESCRIPTION OF DRAWINGS

[0015] Figure 1 Preliminary screening of compound inhibition of ZIKV-NLuc luciferase activity experimental steps; Figure 2 Luciferase expression amount of 5 μM of the compound; Figure 3 EC of the compound TDL-68 50 EC of the compound TDL-28 50 =2.722 μM); Figure 4CC for compound TDL-68 50 CC for compound TDL-28 50 = 216.1 μM); Figure 5 EC for compound TDL-72 50 EC for compound TDL-72 50 = 2.574 μM); Figure 6 CC for compound TDL-72 50 CC for compound TDL-72 50 = 131.4 μM). DETAILED DESCRIPTION

[0016] The application will be further described by the following specific examples. These examples are intended to be illustrative only and are not intended to limit the scope of the application. Various modifications and changes can be made by those skilled in the art to the application disclosed herein, which modifications and changes are intended to fall within the scope of the application as defined by the appended claims. Example 1

[0017] , Compound TDL-68-S1 (0.2 mmol), catalyst Pd(OAc)2(4.48 mg, 10 mol%) and oxidant PhI(OAc)2(128.8 mg, 0.4 mmol) were added into a 25 mL sealed tube, respectively. Under the condition of nitrogen protection, dichloromethane (2 mL), H2O (2 eq) and AcOH (10.0 eq) were added in turn, and the reaction system was mixed uniformly, and then was put into a 50 o C oil bath for reaction for 12 h. After the reaction was completed, the column was spun dry (PE: EA = 2: 1) to obtain compound TDL-68. 1H NMR (400 MHz, CDC13) δ 8.60 - 8.53 (m, 1H), 7.61 (td, J = 7.8, 1.8 Hz, 1H), 7.21 (dd, J = 7.5, 4.9 Hz, 1H), 7.15 (tdd, J = 7.3, 5.1, 1.8 Hz, 1H), 7.06 (d, J = 8.0 Hz, 1H), 6.97 - 6.89 (m, 2H), 6.84 (td, J = 7.6, 1.7 Hz, 1H), 4.81 (s, 1H), 4.47 (d, J = 11.5 Hz, 1H), 4.30 (d, J = 11.6 Hz, 1H), 4.11 (d, J = 11.4 Hz, 1H), 3.92 (d, J = 11.4 Hz, 1H), 3.24 (dd, J = 13.7, 1.5 Hz, 1H), 3.11 (dd, J = 13.7, 1.7 Hz, 1H), 2.00 (s, 3H). 13 C NMR (101 MHz, CDC13) δ 171.1, 162.4, 161.5 (d, J = 246.4 Hz) 148.5, 136.6, 132.6 (d, J = 4.3 Hz), 128.4 (d, J = 8.5 Hz), 123.7, 123.6 (d, J = 13.2 Hz), 122.1, 121.9, 115.3, 115.1. 19 F NMR (377 MHz, CDC13) δ -116.15. HRMS (ESI): m / z calcd. For [C 17 H 18 NO3FNa, M+Na] + :326.1162; found: 326.1162. Example 2

[0018] , Compound TDL-72-S1 (0.2 mmol), catalyst Pd(OAc)2(4.48 mg, 10 mol%) and oxidant PhI(OAc)2(128.8 mg, 0.4 mmol) were added into a 25 mL sealed tube, respectively. Under the condition of nitrogen protection, dichloromethane (2 mL), H2O (2 eq) and AcOH (10.0 eq) were added in turn, the reaction system was mixed uniformly, and then was put into 50o C The reaction was stirred in oil bath for 12 h. After reaction, the column was spin-dried (PE: EA = 2: 1) to get compound TDL-72. 1 H NMR (400 MHz, CDC13) δ 8.56 (ddd, J = 4.9, 1.9, 0.9 Hz, 1H), 7.60 (td, J = 7.8, 1.9 Hz, 1H), 7.28 (dd, J = 7.8, 1.5 Hz, 1H), 7.21 (ddd, J = 7.5, 4.9, 1.0 Hz, 1H), 7.11 (td, J = 7.6, 1.9 Hz, 1H), 7.06 (td, J = 7.5, 1.5 Hz, 1H), 7.01 (dt, J = 8.1, 1.0 Hz, 1H), 6.92 (dd, J = 7.6, 1.8 Hz, 1H), 4.94 (s, 1H), 4.55 (d, J = 11.6 Hz, 1H), 4.31 (d, J = 11.6 Hz, 1H), 4.17 (d, J = 11.5 Hz, 1H), 3.92 (d, J = 11.5 Hz, 1H), 3.41 (d, J = 13.9 Hz, 1H), 3.23 (d, J = 13.9 Hz, 1H), 1.98 (s, 3H). 13 C NMR (101 MHz, CDC13) δ 171.0, 162.4, 148.5, 136.6, 135.3, 134.7, 132.3, 129.5, 128.0, 126.4, 122.2, 122.1, 66.0, 65.4, 49.6, 36.7, 20.8. HRMS (ESI): m / z calcd. For [C 17 H 18 NO3ClNa, M+Na] + : 342.0867; found: 342.0864. Example 3

[0019] This example is a preliminary detection process and results of the compounds TDL-68 and TDL-72 described in Examples 1 and 2 inhibiting Zika virus at the cellular level. The materials and consumables listed in this example can be obtained from commercial channels unless otherwise specified. Cells and viruses come from the cell and microbial resource library of CTCC or other relevant institutions. The experimental methods of this example are standard molecular biology, cell biology or virology procedures that researchers in the field can easily understand and operate. The specific steps are as follows: 1. Cells: Vero (African green monkey kidney cells), culture conditions: DMEM medium 90% + 10% bovine serum + 1% Penicillin-Streptomycin.

[0020] 2. Virus The Zika virus gene sequence and Nano luciferase (NLuc) gene sequence were connected on plasmid pACYC177 using existing conventional genetic means to obtain plasmid pZIKV-Nluc (stored in the laboratory). The cells were inoculated one day in advance so that they reached 70-90% confluence at the time of transfection. Lipofectamine 3000™ was used to prepare plasmid DNA-liposome complexes, and then the DNA-liposome complexes were added to the cells. After 48 h, the supernatant was harvested to infect Vero cells, and the recombinant virus ZIKV-Nluc was obtained (the specific process can be referred to Gao, J.; Chen, J.; Lu, W.; Cai, J.; Shi, L.; Zhao, W.; Zhang, B., Construction of an infectious clone of Zika virus stably expressing an EGFP marker in a eukaryotic expression system. Virology Journal 2021, 18 (1).)。

[0021] 3. Main reagents: DMEM (Gibco, cat: C11995500BT); Fetal bovine serum FBS (Gibco, cat: 10270-106); Plasmid pACYC177 (Mingling Biological, cat: P0995); Double-antibiotic Pen-Strep (10,000 U / mL) (M&C gene biotechnology, cat: G2723M3); Anti-Zika virus positive drug: Ribavirin (Solarbio, cat:IR0090); Gaussia-Lumi™ Luciferase Reporter Gene Assay Kit (Beyotime, cat: RG072S); pZIKV-NLuc transfection reagent Lipofectamine 3000™ (invitrogen, cat: 2319757).

[0022] 4. Experimental steps: Step 1: As Figure 1 As shown, the compound inhibits ZIKV replication as detected: Vero cells were subjected to a concentration of 1.5 × 10⁻⁶. 4 Cells were seeded into 96-well plates and incubated at 37 °C for 12 h until cell confluence reached 90–95%. The original culture medium was discarded, and 100 µL of the test compound solution (dissolved in cell culture medium, final concentration 5 µM) or positive control drug (Ribavirin dissolved in cell culture medium, final concentration 100 µM) was added to each well, with 3 replicates. The plates were incubated at 37 °C for 4 h, the supernatant was discarded, and ZIKV-NLuc virus dilution buffer (MOI = 0.01) and the test compound (final concentration 5 µM) or positive control drug (final concentration 100 µM) were added. The plates were incubated at 37 °C for 2 h, free virus was washed away, and 100 µL of the test compound solution or positive control drug (the compound and positive control drug were dissolved in culture medium) was added to bring the final concentration of the compound to 5 µM and the final concentration of the positive control drug to 100 µM. The plates were incubated at 37 °C for 48 h, and then 100 µL of the test compound solution or positive control drug was added to each well. μL of Beyotime's Gaussia-Lumi™ assay reagent was used to detect Nanoluciferase (NLuc) using a multi-functional microplate reader. The emission wavelength was 485 nm, and the fluorescence value was read. Compound inhibition rate (%) = [1 - (OD compound - OD blank) / (OD virus control - OD blank)] * 100. See details for further information. Figure 2 ,Depend on Figure 2 It is known that TDL-28 and TDL-72 have anti-Zika virus activity at 5 µM.

[0023] Step 2: Replace the 5 µM test compound from Step 1 with test compounds at final concentrations of 0.04 μM, 0.2 μM, 1 μM, 5 μM, 25 μM, and 50 μM. Following Step 1, calculate the relative ZIKV infection luciferase activity as: compound fluorescence value / virus control fluorescence value. Then, based on the relative ZIKV infection luciferase activity, use GraphPad Prism software to determine the EC50 (exhibition coefficient) for inhibiting viral replication. 50 For details, please see the results. Figure 3 andFigure 5 .Depend on Figure 3 It can be seen that the EC of compound TDL-68 50 =2.722 μM, from Figure 5 It can be seen that the EC of compound TDL-72 50 =2.574 μM.

[0024] The experiment included three control groups: Positive drug group: Added virus and added Ribavirin at a final concentration of 100 µM; Virus infection group: only add viruses, no compounds; Normal cell group: No virus or drug added.

[0025] Step 3: MTT assay for cytotoxicity: 5 × 10⁻⁶ 4 Cells were seeded in 96-well cell culture plates containing 100 μL of cell culture medium. 100 μL of a series of compound solutions (dissolved in the medium) were added, resulting in final concentrations of 0.4 μM, 2 μM, 10 μM, 50 μM, 250 μM, and 500 μM, respectively; each concentration was repeated in triplicate. A control without the compound was also included. After 72 h of cell culture, 100 μL of supernatant was discarded, and 20 μL of MTT (5 mg / mL) was added. The cells were incubated at 37 ℃ for 4 h. After centrifugation, 100 μL of supernatant was discarded, and 100 μL of DMSO was added. The cells were incubated at room temperature in the dark, and shaken for 15 minutes until the blue formazene dissolved. Cell viability was measured using a microplate reader with OD595 and OD630 as reference wavelengths. The CC value was calculated based on the cell viability and obtained using GraphPad Prism software. 50 For details, please see the results. Figure 4 , Figure 6 .Depend on Figure 4 It can be seen that the CC of compound TDL-68 50 =216.1 μM, from Figure 6 It can be seen that the CC of compound TDL-72 50 =131.4 μM.

[0026] Step 4: Calculation of the drug's antiviral activity selectivity index: Selective Index (SI) = CC 50 / EC 50 .

[0027] 5. Test Results: Conclusion: Compounds TDL-68 and TDL-72 have significant inhibitory effect on the proliferation of ZIKV-NLuc recombinant virus. The drug antiviral activity selection index SI is 79.39 and 51.05 respectively (the selection index of the control drug Ribavirin is 75).

[0028] The above merely describes preferred specific embodiments of the present application, but the protection scope of the present application is not limited thereto, and any person skilled in the art can make equivalent replacements or changes within the technical range disclosed by the present application according to the technical solution and inventive concept of the present application, which should be covered within the protection scope of the present application.

Claims

1. Isopropylpyridine-derived 1,3-hydroxy / acetoxy compounds, characterized in that, The structural formula is as follows: 。 2. The method for synthesizing the isopropylpyridine-derived 1,3-hydroxy / acetoxy compound as described in claim 1, characterized in that, The method comprises the following steps: adding TDL-S1 into an organic solvent, and reacting in the presence of palladium acetate, iodobenzene acetate, water and acetic acid to obtain an isopropyl pyridine derivative 1,3-hydroxy / acetoxy compound; the reaction equation is as follows: ， R is selected from m-fluorobenzyl or m-chlorobenzyl.

3. The process for the synthesis of isopropylpyridine derived 1,3-hydroxy / acetoxy compounds as claimed in claim 2, wherein: The organic solvent is selected from dichloromethane, chloroform or 1,2-dichloroethane.

4. The process for the synthesis of isopropyl pyridine derived 1,3-hydroxy / acetoxy compounds as claimed in claim 2, wherein: The reaction temperature is 40-60 DEG C.

5. The process for the synthesis of isopropyl pyridine derived 1,3-hydroxy / acetoxy compounds as claimed in claim 2, wherein: The molar ratio of the compound TDL-S1, palladium acetate, iodobenzene acetate, water and acetic acid is 1:0.1:2:2:

10.

6. The use of the isopropyl pyridine derivative 1,3-hydroxy / acetoxy compound according to claim 1 in the preparation of an antiviral drug.

7. The use of the isopropyl pyridine derivative 1,3-hydroxy / acetoxy compound according to claim 6 in the preparation of an antiviral drug, wherein the virus is Zika virus.

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