Preparation method of a paeonol derivative and application of the paeonol derivative in antibacterial and antitumor fields

Abstract

The application discloses a preparation method of a novel paeonol derivative and application of the paeonol derivative in the fields of bacteriostasis and antitumor, and belongs to the technical field of bacteriostatic and antitumor active compounds, and specifically designs a paeonol derivative, a synthesis method of the compound, a pharmaceutical composition containing the paeonol derivative, and application of the paeonol derivative in preparation of bacteriostatic and antitumor preparations. The application provides a series of compounds with bacteriostatic and antitumor activities, the compounds not only have significant bacteriostatic and antitumor effects, but also have the effect of promoting tumor cell apoptosis, and the compounds have novel structures, excellent activities, and good clinical application prospects.

Description

Technical Field

[0001] This invention belongs to the technical field of antibacterial and antitumor active compounds, specifically designing a paeonol derivative, a method for synthesizing the compound, a pharmaceutical composition containing the paeonol derivative, and its application in the preparation of antibacterial and antitumor agents. Background Technology

[0002] The information disclosed in this background section is intended only to enhance understanding of the overall background of the invention and is not necessarily to be construed as an admission or in any way implying that such information constitutes prior art known to those skilled in the art.

[0003] Paeonol, chemically known as 2-hydroxy-4-methoxyacetophenone, also called peony bark ketone, is an active ingredient mainly extracted and isolated from the root bark of Paeonia lactiflora, Paeonia suffruticosa, and Paeonia veitchii. Paeonia suffruticosa root bark, the outer layer of the peony root, is classified under the Paeonia genus and has a history of over 1000 years. Paeonol belongs to the flavonoid derivative class and possesses many significant biological effects, having been used for antibacterial, antitumor, anti-inflammatory, analgesic, antioxidant, antidiabetic, and anti-atherosclerotic purposes. Studies have found that paeonol can also prevent memory loss after ischemic stroke by reducing amyloid precursor protein and β-amyloid precursor protein lyase.

[0004] Paeonol belongs to the small-molecule phenol class and has a simple chemical structure. However, its poor water solubility and short duration of action limit its clinical application, making structural modification essential. Chalcones (1,3-diphenylpropenone) contain two benzene rings and an α,β-enone structure, allowing them to bind to various receptors through covalent bonds, hydrogen bonds, and hydrophobic interactions, exhibiting broad biological activities such as antibacterial, antitumor, and anti-inflammatory effects. Studies have shown that modifying the chalcone structure, such as introducing substituents onto the benzene rings or olefin bonds, or replacing the benzene ring with isosteric compounds, can yield more potent chalcone derivatives. Summary of the Invention

[0005] In response to the needs of existing technologies, the purpose of this invention is to provide a series of compounds with antibacterial and antitumor activities. These compounds not only have significant antibacterial and antitumor effects, but also promote tumor cell apoptosis, and have good prospects for clinical application.

[0006] Specifically, the present invention provides the following technical solution: In a first aspect, the present invention provides a paeonol derivative having the structure shown in Formula I, or an optical isomer, diastereomer, racemic mixture, pharmaceutically acceptable salt, solvate, or prodrug of the compound shown in Formula I. Formula I R1 is selected from hydrogen, hydroxyl, or fluorine; R2 is selected from hydroxyl, hydrogen, methoxy, or ethoxy; and R3 is selected from hydrogen or fluorine.

[0007] In this series of embodiments, Formula I is selected from the following specific compounds: (2-Fluoro-4-hydroxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (code 6a); 3-(2-fluoro-3-hydroxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (code 6b); 3-(2-fluoro-4-methoxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (code name 6c); 3-(2,6-Difluoro-4-hydroxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (code name 6d); 3-(2,6-Difluoro-4-methoxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (code name 6e); 3-(4-ethoxy-2,6-difluorophenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (code name 6f); 3-(2,3-difluoro-4-hydroxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (code 6g).

[0008] Preferably, the pharmaceutically acceptable salt described in the first aspect above should be a non-toxic salt, intended to increase the water solubility and bioavailability of the compound. Further, the pharmaceutically acceptable salt mainly includes quaternary ammonium salts formed by the compound of Formula I with inorganic or organic acids; wherein the inorganic acid is, but is not limited to, hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, diphosphoric acid, hydrobromic acid, hydroiodic acid, and hexafluorophosphate; and the organic acid is, but is not limited to, acetic acid, maleic acid, fumaric acid, tartaric acid, malic acid, succinic acid, lactic acid, p-toluenesulfonic acid, salicylic acid, and oxalic acid.

[0009] Those skilled in the art can select organic and inorganic acids based on solubility, stability, formulation process, etc., to obtain the corresponding salts. The determination and optimization of the salts are within the experience range of those skilled in the art.

[0010] In a second aspect, the present invention provides a method for synthesizing the paeonol derivatives described in the first aspect, using paeonol (1) as the starting material, and the synthetic route is as follows:

[0011] Preferably, in the above synthesis method, the specific synthesis method of step a is as follows: Paeonol, thionyl chloride and benzaldehyde containing R1, R2 and R3 substituents are dissolved in EtOH and reacted overnight at room temperature to obtain the compound shown in Formula I.

[0012] Furthermore, the molar ratio of paeonol, benzaldehyde containing R1, R2 and R3 substituents, and thionyl chloride is 1:1~1.2:1~2.84.

[0013] Furthermore, the amount of EtOH used is 4-6 mL of EtOH per millimole of paeonol.

[0014] In a third aspect, the present invention provides a pharmaceutical composition comprising the paeonol derivatives described in the first aspect, and further comprising a pharmaceutically necessary carrier.

[0015] In the above-mentioned pharmaceutical composition, the paeonol derivatives mentioned in the first aspect should be at a therapeutically effective dose, and the effective dose can be routinely adjusted according to the purpose of the clinical application of the pharmaceutical composition.

[0016] Preferably, the pharmaceutical composition is an oral or parenteral preparation. In specific examples, the pharmaceutical composition is a tablet, pill, capsule, or injection.

[0017] In a fourth aspect, the present invention provides the use of the paeonol derivatives described in the first aspect and the pharmaceutical composition described in the third aspect in the preparation of antibacterial agents.

[0018] Preferably, the antibacterial agent is used against, but is not limited to, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Klebsiella pneumoniae, Staphylococcus aureus, and Enterobacter cholerae.

[0019] In a fifth aspect, the present invention provides the use of the paeonol derivatives described in the first aspect and the pharmaceutical composition described in the third aspect in the preparation of antitumor agents.

[0020] Preferably, the antitumor agent is used for one of the following cancers: cervical cancer, liver cancer, breast cancer, skin cancer, head and neck cancer, lung cancer, esophageal cancer, pancreatic cancer, colon cancer, kidney cancer, ureteral cancer, and bladder cancer.

[0021] In a sixth aspect, the present invention provides a treatment method for tumor diseases, the treatment method comprising administering to an individual in need the pharmaceutical composition comprising the paeonol derivatives of the first aspect and the pharmaceutical composition of the third aspect.

[0022] Preferably, the method of use includes, but is not limited to, delivering the pharmaceutical composition containing paeonol derivatives (as described in the first aspect) and the pharmaceutical composition described in the third aspect to the lesion site via oral administration, injection, or interventional procedures.

[0023] The beneficial effects of one or more of the above technical solutions are: This invention synthesizes a series of paeonol derivatives with antibacterial and antitumor activities. These compounds exhibit good activity in in vitro antitumor proliferation experiments, and their in vitro inhibitory effect on tumor cell proliferation is comparable to that of the positive control drug cisplatin. Therefore, they may become drugs with high therapeutic index that can be used clinically. Attached Figure Description

[0024] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an improper limitation of the invention.

[0025] Figure 1 The diagram shows the spatial conformation of compound 6d and its binding mode with PI3K. Detailed Implementation

[0026] It should be noted that the following detailed descriptions are exemplary and intended to provide further illustration of the invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0027] It should be noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of exemplary embodiments according to the invention. As used herein, the singular form is intended to include the plural form as well, unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components, and / or combinations thereof.

[0028] To enable those skilled in the art to better understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0029] Example 1: Preparation of the compound The compound of formula I described in this invention can be prepared via the following reaction route:

[0030] The reagents used in the above preparation process are: a: thionyl chloride; ethanol; room temperature; 12 h.

[0031] Specifically, the present invention provides the preparation process of the illustrative compounds and the effect verification data in the embodiments.

[0032] (1) Preparation of 3-(2-fluoro-4-hydroxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (6a) 0.50 g (3.01 mmol) of paeonol and 0.51 g (3.61 mmol) of 2-fluoro-4-hydroxybenzaldehyde were placed in a round-bottom flask, and 15 mL of anhydrous ethanol was added. Then, 1.02 g (8.55 mmol) of thionyl chloride was slowly added dropwise. After the addition was complete, the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction was stopped, and 50 mL of saturated sodium chloride solution was added. The mixture was extracted three times with ethyl acetate (50 mL × 3). The organic phases were combined, dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under vacuum. The filtrate was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 9:1) to give 0.28 g of the target compound 6a as a yellow powder. The yield was 32%, and the mp values ​​were 190–192 °C. 1 H NMR (600 MHz, DMSO- d6 ): δ 13.49 (s, 1H, OH), 10.67 (s, 1H, OH), 8.20 (d, J = 9.1 Hz, 1H, CH), 8.00 (t, J = 8.8 Hz, 1H, CH), 7.92-7.82 (m, 2H,ArH), 6.77-6.63 (m, 2H, ArH), 6.62-6.47 (m, 2H, ArH), 3.85 (s, 3H, CH3); 13 CNMR (151 MHz, DMSO- d6 ): δ 192.05, 166.39, 166.18, 163.63, 161.97, 136.33,132.87, 130.91, 119.66, 114.28, 113.21, 107.91, 103.39, 103.23, 101.42,56.23; MS (ESI) m / z : calcd. for C 16 H 13 FO4[M+H] + : 289.08, found: 289.00. (2) Preparation of 3-(2-fluoro-3-hydroxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (6b) 0.50 g (3.01 mmol) of paeonol and 0.51 g (3.61 mmol) of 2-fluoro-3-hydroxybenzaldehyde were placed in a round-bottom flask, 15 mL of anhydrous ethanol was added, and then 1.02 g (8.55 mmol) of thionyl chloride was slowly added dropwise. After the addition was complete, the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction was stopped, 50 mL of saturated sodium chloride solution was added, and the mixture was extracted three times with ethyl acetate (50 mL × 3). The organic phases were combined, dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under vacuum. The filtrate was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 15:1) to give 0.33 g of yellow powdery solid target compound 6b. Yield 38%, mp 152–154 °C. 1 H NMR (600 MHz, DMSO- d6 ): δ 12.61 (s, 1H, OH), 9.77 (s, 1H, OH), 8.23 ​​(d,J = 9.1 Hz, 1H, CH), 7.78 (s, 1H, CH), 7.12 -7.04 (m, 1H, ArH), 6.93 (t, J =8.1 Hz, 1H, 3.79 (s, 3H, CH3); 13 C NMR (151 MHz, DMSO- d6 ): δ 203.26, 191.95, 166.15,163.99, 163.11, 133.20, 129.94, 113.53, 108.07, 101.45, 99.75, 72.29, 57.07,56.66, 56.29, 27.27; MS (ESI) m / z : calcd. for C 16 H 13 FO4[M+H] + : 289.08, found:289.16. (3) Preparation of 3-(2-fluoro-4-methoxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (6c) 0.50 g (3.01 mmol) of paeonol and 0.56 g (3.61 mmol) of 2-fluoro-4-methoxybenzaldehyde were placed in a round-bottom flask, and 15 mL of anhydrous ethanol was added. Then, 1.02 g (8.55 mmol) of thionyl chloride was slowly added dropwise. After the addition was complete, the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction was stopped, and 50 mL of saturated sodium chloride solution was added. The mixture was extracted three times with ethyl acetate (50 mL × 3). The organic phases were combined, dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under vacuum. The filtrate was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 12:1) to give 0.32 g of the target compound 6c as a yellow powder. Yield: 35%, mp: 146–148 °C. 1 H NMR (600 MHz, DMSO- d6 ): δ 13.44 (s, 1H, OH), 8.22 (d, J = 9.1 Hz, 1H,CH), 8.11 (t, J = 8.8 Hz, 1H, CH), 7.96-7.84 (m, 2H, ArH), 7.00-6.89 (m, 2H,ArH), 6.60-6.49 (m, 2H, ArH), 3.85 (s, 6H, CH3); 13 C NMR (151 MHz, DMSO- d6 ): δ191.99, 166.47, 166.20, 163.48, 161.85, 135.80, 132.99, 130.64, 120.76,114.29, 112.15, 107.95, 102.21, 102.04, 101.41, 56.51, 56.25; MS (ESI) m / z :calcd. for C 17 H 17 FO4[M+H] + 303.10, found: 303.17. (4) Synthesis of 3-(2,6-difluoro-4-hydroxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (6d) 0.50 g (3.01 mmol) of paeonol and 0.57 g (3.61 mmol) of 2,6-difluoro-4-hydroxybenzaldehyde were placed in a round-bottom flask, and 15 mL of anhydrous ethanol was added. Then, 1.02 g (8.55 mmol) of thionyl chloride was slowly added dropwise. After the addition was complete, the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction was stopped, and 50 mL of saturated sodium chloride solution was added. The mixture was extracted three times (50 mL × 3) with ethyl acetate. The organic phases were combined, dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under vacuum. The filtrate was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 8:1) to give 0.28 g of the target compound 6d as a yellow powder. Yield: 30%, mp: 205–207 °C. 1 H NMR (600 MHz, DMSO- d6 ): δ 13.16 (s, 1H, OH), 7.93 (d, J = 9.0 Hz, 1H,OH), 7.73 (s, 2H, CH), 6.62 (d, J = 11.3 Hz, 2H, ArH), 6.58-6.50 (m, 3H,ArH), 3.84 (s, 3H, CH3); 13 C NMR (151 MHz, DMSO- d6 ): δ 191.69, 166.40, 166.04,165.79, 132.46, 130.41, 128.66, 122.87, 114.42, 110.74, 108.14, 101.51,101.33, 100.64, 100.46, 56.23; MS (ESI) m / z : calcd. for C 16 H 12 F2O4[M+H] + :307.07, found: 307.35. (5) Synthesis of 3-(2,6-difluoro-4-methoxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (6e) 0.50 g (3.01 mmol) of paeonol and 0.62 g (3.61 mmol) of 2,6-difluoro-4-methoxybenzaldehyde were placed in a round-bottom flask, and 15 mL of anhydrous ethanol was added. Then, 1.02 g (8.55 mmol) of thionyl chloride was slowly added dropwise. After the addition was complete, the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction was stopped, and 50 mL of saturated sodium chloride solution was added. The mixture was extracted three times (50 mL × 3) with ethyl acetate. The organic phases were combined, dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under vacuum. The filtrate was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 12:1) to give 0.38 g of the target compound 6e as a yellow powder. The yield was 39%, and the mp values ​​were 173–175 °C. 1 H NMR (600 MHz, DMSO- d6 ): δ 13.09 (s, 1H, OH), 7.96 (d, J = 9.2 Hz,1H, CH), 7.83-7.71 (m, 2H, ArH), 6.94 (d, J = 10.9 Hz, 2H, ArH), 6.58 (dd, J= 8.9, 2.5 Hz, 1H, CH), 6.53 (d, J = 2.5 Hz, 1H, ArH), 3.87 (s, 3H, CH3), 3.85 (s, 3H, CH3); 13 C NMR (151 MHz, DMSO- d6 ): δ 191.60, 166.47, 165.74,132.63, 129.95, 124.17, 114.49, 108.21, 105.06, 101.52, 99.79, 99.60, 57.05,56.26, 31.62, 30.82, 30.28; MS (ESI) m / z : calcd. for C 17 H 14 F2O4[M+H] + 321.09, found: 321.15. (6) Synthesis of 3-(4-ethoxy-2,6-difluorophenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (6f) 0.50 g (3.01 mmol) of paeonol and 0.67 g (3.61 mmol) of 2,6-difluoro-4-ethoxybenzaldehyde were placed in a round-bottom flask, and 15 mL of anhydrous ethanol was added. Then, 1.02 g (8.55 mmol) of thionyl chloride was slowly added dropwise. After the addition was complete, the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction was stopped, and 50 mL of saturated sodium chloride solution was added. The mixture was extracted three times (50 mL × 3) with ethyl acetate. The organic phases were combined, dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under vacuum. The filtrate was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 10:1) to give 0.31 g of the target compound 6f as a yellow powder. The yield was 31%, and the mp values ​​were 134–136 °C. 1 H NMR (600 MHz, DMSO- d6 ): δ 13.10 (s, 1H, OH), 7.95 (d, J = 9.1 Hz,1H, CH), 7.82-7.69 (m, 2H, ArH, CH), 6.91 (d, J = 11.0 Hz, 2H, ArH), 6.61-6.49 (m, 2H, ArH), 4.14 (q, J = 7.0 Hz, 2H, CH2), 3.85 (s, 3H, CH3), 1.34 (t,J = 7.0 Hz, 3H, CH3); 13 C NMR (151 MHz, DMSO- d6 ): δ 191.60, 166.46, 165.75,163.55, 162.36, 161.81, 132.60, 129.98, 123.96, 114.47, 108.20, 104.92,101.52, 100.05, 99.86, 65.27, 56.26, 14.71; MS (ESI) m / z : calcd. forC 18 H 16 F2O4[M+H] + : 335.10, found: 335.15. (7) Preparation of 3-(2,3-difluoro-4-hydroxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (6g) 0.50 g (3.01 mmol) of paeonol and 0.57 g (3.61 mmol) of 2,3-difluoro-4-hydroxybenzaldehyde were placed in a round-bottom flask, and 15 mL of anhydrous ethanol was added. Then, 1.02 g (8.55 mmol) of thionyl chloride was slowly added dropwise. After the addition was complete, the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction was stopped, and 50 mL of saturated sodium chloride solution was added. The mixture was extracted three times with ethyl acetate (50 mL × 3). The organic phases were combined, dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under vacuum. The filtrate was then subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 14:1) to give 6 g of the target compound as a yellow powder. The yield was 34%, and the mp values ​​were 179–181 °C. 1 H NMR (600 MHz, DMSO- d6 ): δ 13.37 (s, 1H, OH), 8.19 (d, J = 9.1 Hz, 1H,OH), 7.90 (d, J = 15.5 Hz, 1H, CH), 7.84-7.77 (m, 2H, ArH), 6.91-6.82 (m, 2H,ArH), 6.68 (dd, J = 8.8, 2.4 Hz, 1H, CH), 6.57 (dd, J = 9.0, 2.6 Hz, 1H,ArH), 6.52 (d, J = 2.6 Hz, 1H, CH), 3.85 (s, 3H, CH3); 13 C NMR (151 MHz, DMSO- d6 ): δ 191.79, 190.20, 166.51, 166.16, 163.41, 135.34, 132.97, 128.56,124.32, 121.36, 114.28, 110.66, 108.00, 101.43, 73.89, 56.26; MS (ESI) m / z :calcd. for C 16 H 12 F2O4[MH] - : 305.07, found: 305.91. Example 2: Detection of the in vitro antibacterial activity of the compound (1) Experimental materials: Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), Candida albicans (ATCC 10231), Klebsiella pneumoniae (ATCC 700603), Staphylococcus aureus (ATCC 25923) and Enterobacter hominis (ATCC 700323), cefazolin, cefoperazone, amphotericin B.

[0033] (2) Experimental Methods: Accurately weigh 10 g tryptone, 5 g yeast extract, and 10 g sodium chloride, dissolve them in 950 mL of pure water, stir until fully dissolved, and bring the volume to 1000 mL. Dispense the solution into 50 mL Erlenmeyer flasks. Seal the flasks with heat-resistant tissue culture sealing film, place them in an autoclave, and sterilize at 121 °C for 30 min. After cooling, store at room temperature for later use. In a biosafety cabinet, take 500 µL of the prepared glycerol culture and add it to an Erlenmeyer flask containing 50 mL of sterilized LB liquid medium. Shake well, seal the flask, and incubate overnight in a shaker at 37 °C. The cultured strains in the logarithmic growth phase (OD2) were then... 600 nm = 0.8~1.3) Dilute with LB liquid medium to a final concentration of 1×10 6 CFU / mL for later use. Dissolve the target compound in DMSO to prepare a solution of 9 × 10⁻⁶ CFU / mL. 4 µmol·L -1 The stock solution was then diluted with LB liquid medium to 60 µmol·L⁻¹. -1 Ensure the DMSO concentration does not exceed 0.1%. Perform the following experiments in sterilized 1.5 mL EP tubes: Negative control group: 0.5 mL bacterial suspension + 0.5 mL culture medium; Negative control group: 0.5 mL culture medium + 0.5 mL culture medium; Experimental group: 0.5 mL bacterial suspension + 0.5 mL target compound; Experimental control group: 0.5 mL culture medium + 0.5 mL target compound; Positive control group: 0.5 mL bacterial suspension + 0.5 mL positive control agent; Positive control group: 0.5 mL culture medium + 0.5 mL positive control agent. Ensure the final concentration of the bacterial suspension is 5 × 10⁻⁶. 5 CFU / mL, the final concentration of the target compound was 30 µmol·L⁻¹. -1 After incubating the EP tubes in a shaker at 37 ℃ for 24 h, the OD value at 600 nm was measured using a multi-functional microplate reader. The experiment was repeated three times, and the average value was used to calculate the inhibition rate.

[0034] (3) Calculate the inhibition rate: Inhibition rate T = (AB) / A × 100%, where A = OD value of negative group - OD value of negative blank group, and B = OD value of experimental group - OD value of experimental blank group or OD value of positive group - OD value of positive blank group. Experimental results: Compounds 6a, 6b, 6d, 6e and 6g were tested at 30 µmol·L⁻¹. -1 The inhibition rates against Candida albicans at the specified concentrations were 99.45%, 85.81%, 95.76%, 92.91%, and 98.38%, respectively, comparable to the inhibition rate of the positive control drug amphotericin B against Candida albicans; compounds 6a, 6d, and 6g at 30 µmol·L⁻¹... -1 The inhibition rates against Staphylococcus aureus at the specified concentrations were 99.36%, 99.88%, and 80.44%, respectively, showing good antibacterial effects.

[0035] Table 1

[0036] Example 3: Detection of the in vitro antitumor activity of the compound (1) Experimental materials: cisplatin, human cervical cancer cell line (Hela), human liver cancer cell line (HepG-2), human breast cancer cell line (MCF-7), and human embryonic kidney cell line (293T).

[0037] (2) Experimental Methods: Cells were cultured in cell culture flasks until the logarithmic growth phase. The culture medium was aspirated from the flasks, and the cells were gently rinsed 1-2 times with 1 mL of PBS to remove residual serum and metabolites. 1 mL of trypsin was added for 1-2 min, followed by 2 mL of 10% culture medium to stop the digestion. The cells were gently pipetted until all corners were clean. The cell suspension was transferred to a 15 mL centrifuge tube and centrifuged at 1000 rpm for 5 min. The supernatant was quickly aspirated, and 1 mL of 10% culture medium was added and mixed thoroughly to prepare a single-cell suspension. The single-cell suspension was diluted, and 100 μL of the suspension (approximately 5000 cells) was added to each well and evenly seeded into 96-well plates. No cells were added to the control group. PBS was placed around the 96-well plates to reduce the impact of evaporation. The 96-well plates were pre-cultured at 37 ℃ in a 5% CO2 incubator for 24 h to allow the cells to adhere. The original culture medium was aspirated. The negative control group was given 2% culture medium, while the experimental groups were given 90, 60, 40, 26.67, 17.78, 11.85, and 7.90 µmol·L⁻¹, respectively. -1 The drug was administered in triplicate for each concentration, and cultured for 24 h. The culture medium and different concentrations of drug were aspirated, and 100 μL of medium containing 10% CCK-8 (1 mL CCK8 + 9 mL 2% medium) was added to each well. The wells were incubated for about 1 h, and the OD values ​​of each well were measured at 450 nm using a multi-functional microplate reader. The data were then processed.

[0038] (3) Calculate the inhibition rate: Inhibition rate = (OD value of negative control group - OD value of experimental group) / (OD value of negative control group - OD value of blank group) × 100%. Process the calculated inhibition rate using GraphPad Prism 8.0 software. 50 The logarithm (inhibition rate) and response-variable slope (four-parameter) formulas were used for fitting, and the results were expressed as mean ± standard deviation (x ± s).

[0039] Experimental results: Compounds 6a, 6b, 6d, and 6g exhibited certain anti-proliferative activity against HeLa cells, with an IC50 value of [missing value]. 50 The concentrations were 48.74 ± 2.50 µmol·L⁻¹. -1 161.70 ± 16.18 µmol·L -1 25.19 ± 0.49 µmol·L -1 and 55.62 ± 4.27 µmol·L -1 Compound 6d showed superior antiproliferative activity against HeLa cells compared to the positive control cisplatin (IC50). 50 = 29.10 ± 1.74 µmol·L -1 Compounds 6a, 6d, and 6g showed their effects on the IC50 values ​​of HepG-2 cells. 50 The values ​​were 40.57 ± 2.53 µmol·L⁻¹. -1 29.52 ± 1.19 µmol·L -1 and 48.76 ± 1.90 µmol·L -1 Superior to the positive control drug cisplatin (IC50) 50 = 95.12 ± 2.04 µmol·L -1 In MCF-7 cells, compounds 6a, 6b, 6d, 6f, and 6g exhibited antiproliferative activity, with an IC50 value of [missing value]. 50 The concentrations were 46.38 ± 2.38 µmol·L⁻¹. -1 93.24 ± 0.65 µmol·L -1 31.01 ± 1.15 µmol·L -1 162.00 ± 1.52 µmol·L -1 and 52.53 ± 3.75 µmol·L -1 Among them, compounds 6a and 6g showed antiproliferative activity similar to that of the positive control drug cisplatin (IC50). 50 = 56.64 ± 21.39 µmol·L -1The antiproliferative activity of compound 6d was comparable to that of the positive control cisplatin. Regarding cytotoxicity, compounds 6c, 6e, 6f, and 6g all exhibited low cytotoxicity, with IC50 values ​​of [missing value]. 50 At 200 µmol·L -1 The above is comparable to the starting material paeonol and the positive control drug cisplatin.

[0040] Table 2

[0041] Example 4 Molecular docking of compounds Experimental Principle: Paeonol and its derivatives induce tumor cell apoptosis by inhibiting the PI3K / AKT and NF-κB signaling pathways; and inhibit tumor cell invasion and metastasis by inhibiting the TGF-β / Smad signaling pathway and inflammation-related pathways such as STAT3 and NF-κB. Activation of the PI3K / AKT signaling pathway is closely related to tumorigenesis and development. In MCF-7 cells, paeonol reduces the expression of PI3K / AKT mRNA, decreases PI3K / AKT protein expression, inhibits the phosphorylation or activation of intracellular messengers, thereby increasing apoptosis and inhibiting cell growth.

[0042] Experimental procedure: The 3D structure of phosphatidylinositol 3-kinase PI3K (PDB ID: 3APC) was downloaded from the PDB database (https: / / www.rcsb.org / ), imported into Discovery Studio 2020 software, the original ligand and solvent molecules were removed, and hydrogenation was performed to obtain the pretreated protein receptor; then compound 6d was imported, and the most suitable active site was selected to dock the receptor with compound 6d.

[0043] Results analysis: The hydroxyl group on the left benzene ring of compound 6d forms a hydrogen bond with GLU880 of PI3K, and the hydroxyl group on the right benzene ring forms a π-electron donor hydrogen bond with HIS658 of PI3K; the two benzene rings form π-cation interactions with ARG849 and ARG690, respectively. Notably, the fluorine atom of compound 6d forms a halogen bond with LEU657, which may be one of the reasons why the target compound 6d exhibits high in vitro antitumor activity.

[0044] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, wherein, R1 is selected from hydrogen, hydroxyl, or fluorine; R2 is selected from hydroxyl, hydrogen, methoxy, or ethoxy; R3 is selected from hydrogen or fluorine.

2. The compound according to claim 1, characterized in that, R1 is hydrogen, hydroxyl, or fluorine.

3. The compound according to claim 1, characterized in that, R2 is a hydroxyl group, hydrogen, methoxy group, or ethoxy group.

4. The compound according to claim 1, characterized in that, R3 is either hydrogen or fluorine.

5. The compound according to claim 1, characterized in that, The compound is: 3-(2-fluoro-4-hydroxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (code name 6a); 3-(2-fluoro-3-hydroxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (code 6b); 3-(2-fluoro-4-methoxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (code name 6c); 3-(2,6-Difluoro-4-hydroxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (code name 6d); 3-(2,6-Difluoro-4-methoxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (code name 6e); 3-(4-ethoxy-2,6-difluorophenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (code name 6f); 3-(2,3-difluoro-4-hydroxyphenyl)-1-(2-hydroxy-4-methoxyphenyl)prop-2-en-1-one (code 6g).

6. A method for preparing the compound according to any one of claims 1 to 5, characterized in that, The process includes the following steps: dissolving paeonol, thionyl chloride, and benzaldehyde containing R1, R2, and R3 substituents in EtOH, and reacting overnight at room temperature to obtain the compound shown in Formula I. Further, the molar ratio of paeonol, benzaldehyde containing R1, R2, and R3 substituents, and thionyl chloride is 1:1 to 1.2:1 to 2.

84. Further, the amount of EtOH used is 4 to 6 mL per millimole of paeonol.

7. The use of any of the compounds according to claims 1 to 5 in the preparation of antibacterial or antitumor drugs.

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