Application of halogenated flavone compound in resisting staphylococcus aureus
The antibacterial ability of flavonoids is enhanced through the transformation of halogenated flavonoids, and the treatment problem of multidrug-resistant bacteria is solved, especially the significant inhibitory effect on Staphylococcus aureus and methicillin-resistant Staphylococcus aureus.
Patent Information
- Application Number
- CN202510273504.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-08
- Publication Date
- 2025-07-22
- Estimated Expiration
- Not applicable · inactive patent
AI Technical Summary
Existing antibiotics face the challenge of multidrug-resistant bacteria, especially infectious diseases such as Staphylococcus aureus and methicillin-resistant Staphylococcus aureus, lack effective treatments.
A class of halogenated flavonoid compounds were developed to modify the structure of flavonoid compounds through halogen atoms, enhance their binding affinity and membrane permeability with bacteria, and prepare antibacterial agents for the treatment of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus.
Halogenated flavonoid compounds show excellent inhibitory activity, especially show significant inhibitory effects on Staphylococcus aureus and methicillin-resistant Staphylococcus aureus, with good application prospects.
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Abstract
Description
Technical Field
[0001] The present invention belongs to the field of biology, and specifically relates to the application of a class of halogenated flavone compounds as anti-Staphylococcus aureus agents. Background Art
[0002] The discovery of penicillin opened the door to the golden age of antibiotics. Since then, different structural types of antibiotics have been discovered one after another. As of 2025, 13 major categories of antibiotics have been discovered, such as β-lactams, quinolones, and aminoglycosides. However, due to the abuse of antibiotics and the adverse toxic and side effects of broad-spectrum antibiotics, multi-drug resistant bacteria will sweep the globe. Flavonoid compounds are mainly a kind of polyphenolic compounds isolated from plants. Structurally, they usually consist of a 15-carbon skeleton and are connected by a 3-carbon bridge to two aromatic rings. So far, more than 10,000 flavonoid natural products have been discovered in nature, and they exhibit significant antibacterial, anti-inflammatory, antioxidant, antiviral, lipid-lowering and other activities. The antibacterial effect of flavonoid compounds is usually direct antibacterial, synergistic antibacterial with antibiotics, and acting on the virulence factors of bacteria.
[0003] Halogen atoms play an important role in drug development. Since the discovery of chloramphenicol in 1947, halogen atoms have become the main components of several types of antibiotics and antibiotic scaffolds. In antibacterial drugs used clinically, halogen atoms are widespread and play an important antibacterial activity role. More than 25% of the marketed drugs and nearly 40% of the active lead compounds contain halogens; among the 50 drugs approved by the US Food and Drug Administration (FDA) in 2021, 14 contain halogens, all indicating the importance of halogen atoms in several types of antibiotics and antibacterial scaffolds. Halogenated compounds can form multiple non-covalent interactions with ligands, thereby affecting the binding affinity, membrane permeability and lipophilicity of the compounds, and further enhancing the antibacterial ability and anti-drug resistance.
[0004] Therefore, the development of flavonoid compounds containing halogen atoms is an important way to develop new antibiotic drugs. Summary of the Invention
[0005] Only some aspects of the present invention are generally described below and are not limited thereto. These aspects and other parts are more fully described later. All references in this specification are incorporated herein by reference in their entirety. When there are differences between the disclosure of this specification and the cited literature, the disclosure of this specification shall prevail.
[0006] The present invention provides a class of halogenated flavone compounds, their preparation methods and their use as anti-Staphylococcus aureus agents. Specifically, the present invention relates to compounds 1-4 of formula I structure, or pharmaceutically acceptable salts thereof, or solvates of salts thereof. The present invention also discloses the use of such compounds for the preparation of anti-Staphylococcus aureus and anti-methicillin-resistant Staphylococcus aureus. The compounds of the present invention have excellent inhibitory activity against Staphylococcus aureus and anti-methicillin-resistant Staphylococcus aureus, and have good application prospects.
[0007] Specifically:
[0008] The present invention relates to a compound of formula I structure, or a pharmaceutically acceptable salt thereof, or a solvate of a salt thereof, characterized in that the compound has the following structure:
[0009] Wherein, X1 and X2 are each independently optionally H, Cl, Br, I; specifically,
[0010] Another embodiment of the present invention provides the use of a class of halogenated flavones as antibacterial agents, characterized in that it comprises a compound of formula I or a pharmaceutically acceptable salt thereof as an active ingredient. In addition, another embodiment of the present invention provides that the same class of halogenated flavones do not have antibacterial activity, characterized in that it contains compounds c-g, specifically
[0011]
[0012] The term "pharmaceutically acceptable salt" in the present invention refers to non-toxic addition salts of inorganic or organic acids and / or bases; see "Salt selection for basic drugs", Int. J. Pharm. 1986, 33, 201–217. Specific embodiments
[0013] For the convenience of further understanding of the present invention, the following examples provide more detailed descriptions thereof; however, these examples are only for better understanding of the invention and are not used to limit the scope or implementation principles of the present invention. The implementation modes of the present invention are not limited to the following content.
[0014] Example 1: Compound 1
[0015]
[0016] Dissolve dechlorochlorflavonin (hereinafter referred to as "Compound a") (30 mg, 1.0 eq) in a reaction sealed tube with ethyl acetate. Pipette DMSO (7.2 μL, 1.2 eq) and hydrobromic acid (HBr, 18.0 μL, 1.8 eq) into the reaction sealed tube. React the mixed reaction solution in an oil bath at 60 °C for 2 h, and monitor the reaction by TLC. After the reaction solution is cooled to room temperature, concentrate it under reduced pressure, and separate and purify it by normal-phase silica gel column chromatography to obtain a yellow solid with a yield of 34%; 1 H NMR (400 MHz, CDCl3) δ 12.68 (s, 1H), 7.87 (d, J = 2.5 Hz, 1H), 7.85 (s, 1H), 7.59 (dd, J = 8.8, 2.5 Hz, 1H), 7.02 (d, J = 8.8 Hz, 1H), 4.10 (s, 3H), 3.94 (s, 3H), 3.93 (s, 3H). 13 C NMR (100 MHz, CDCl3) δ 177.4, 156.8, 154.9, 154.8, 153.8, 149.0, 138.0, 136.6, 134.0, 131.9, 122.0, 119.7, 113.3, 107.8, 100.0, 62.7, 62.4, 61.7. HRESIMS m / z 500.9168 [M + H] + (calcd for C 18 H 16 O7Br2 + , 500.9179);
[0017] Example 2: Compound 2
[0018]
[0019] Weigh N-bromosuccinimide (NBS; 31.0 mg, 3 eq) into a reaction flask. Use dichloromethane (5 mL) as the reaction solvent, add a trace amount of DMSO (0.2 eq) as a catalyst and mix well. Finally, add Compound a (20 mg, 1 eq). Stir the reaction solution at room temperature and monitor the reaction by TLC. After the reaction is completed, add water to terminate the reaction, extract the reaction solution with dichloromethane three times, combine the organic phases, concentrate under reduced pressure, and separate and purify by normal-phase silica gel column chromatography to obtain a yellow solid with a yield of 82%; 1 H NMR (400 MHz, DMSO-d6) δ 12.96 (s, 1H), 10.51 (s, 1H), 7.99 (d, J = 2.4 Hz, 1H), 7.68 (d, J = 2.4 Hz, 1H), 4.01 (s, 3H), 3.81 (s, 3H), 3.75 (s, 3H). 1313C NMR (100 MHz, DMSO-d6) δ 178.3, 155.2, 154.5, 152.7, 151.6, 149.0, 140.2, 137.2, 133.2, 132.3, 120.5, 113.1, 110.4, 108.0, 97.8, 62.0, 61.5, 60.3. HRESIMS m / z 578.8282 [M+H] + (calcd for C 18 H 16 O7Br3 + , 578.8284).
[0020] Example 3: Compound 3
[0021]
[0022] Weigh N-iodosuccinimide (NIS; 35.7 mg, 3 eq) into a reaction flask. Using dichloromethane (5 mL) as the reaction solvent, add a trace amount of DMSO (0.2 eq) as a catalyst and mix well. Finally, add chlorflavonin (hereinafter all referred to as "Compound b") (20 mg, 1 eq). The reaction solution is stirred at room temperature, and the reaction is monitored by TLC. After the reaction is completed, add water to terminate the reaction, extract the reaction solution with dichloromethane three times, combine the organic phases, concentrate under reduced pressure, and separate and purify by normal-phase silica gel column chromatography to obtain a yellow solid with a yield of 82%; 1 1H NMR (400 MHz, CDCl3) δ 12.93 (s, 1H), 7.95–7.83 (m, 2H), 7.74 (s, 1H), 4.08 (s, 3H), 3.94 (s, 3H), 3.91 (s, 3H). 13 13C NMR (100 MHz, CDCl3) δ 177.4, 158.8, 156.4, 153.5, 151.1, 141.1, 137.0, 133.2, 125.3, 124.6, 124.1, 121.1, 114.2, 107.7, 81.6, 62.5, 62.3, 61.6. HRESIMS m / z 630.8507 [M+H] + (calcd for C 18 H 14 O7ClI2 + , 630.8512).
[0023] Example 4: Compound 4
[0024]
[0025] Weigh NIS (38.8 mg, 3 eq) into a reaction flask. Using dichloromethane (5 mL) as the reaction solvent, add a trace amount of DMSO (0.2 eq) as a catalyst and mix well. Finally, add compound a (20 mg, 1 eq). Stir the reaction solution at room temperature, and monitor the reaction progress by TLC. After the reaction is completed, add water to terminate the reaction. Extract the reaction solution with dichloromethane three times, combine the organic phases, concentrate under reduced pressure, and separate and purify by normal-phase silica gel column chromatography to obtain a yellow solid with a yield of 80%; HRESIMS m / z 722.7860 [M+H] + (calcd for C 18 H 14 O7I3 + ,722.7868).
[0026] Example 5: Compound c
[0027]
[0028] Dissolve compound a (20 mg, 1.0 eq) in dichloromethane (4 mL), add NCS (11.6 mg, 1.2 eq), stir the reaction solution at room temperature, and monitor the reaction progress by TLC. After the reaction is completed, add water to terminate the reaction. Extract the reaction solution with dichloromethane three times, combine the organic phases, concentrate under reduced pressure, and separate and purify by normal-phase silica gel column chromatography to obtain a yellow solid with a yield of 53%. 1 HNMR(400MHz,DMSO-d6)δ12.97(s,1H),10.17(s,1H),7.46–7.38(m,2H),7.02(dd,J=8.4,1.1Hz,1H),6.96(td,J=7.5,1.1Hz,1H),4.02(s,3H),3.84(s,3H),3.74(s,3H). 13 C NMR(100MHz,DMSO-d6)δ178.3,157.9,155.7,154.1,151.6,148.1,139.6,133.1,132.4,130.6,118.9,117.1,116.4,107.6,107.4,62.0,61.6,60.1.HRESIMS m / z 379.0583[M+H] + (calcdfor C 18 H 16 O7Cl + ,379.0579).
[0029] Example 6: Compound d
[0030]
[0031] Weigh NBS (15.5 mg, 1.5 eq) into a reaction flask. Using ethanol (3 mL) as the reaction solvent, add a trace amount of DMSO (0.2 eq) as a catalyst and mix well. Finally, add compound a (20 mg, 1.0 eq). Stir the reaction solution at room temperature, and monitor the reaction by TLC. After the reaction is completed, add water to terminate the reaction. Extract the reaction solution with dichloromethane three times, combine the organic phases, concentrate under reduced pressure, and separate and purify by normal-phase silica gel column chromatography to obtain a yellow solid with a yield of 83%. 1 1H NMR (400 MHz, CDCl3) δ 12.80 (s, 1H), 7.83 (s, 1H), 7.78 (dd, J = 8.2, 1.7 Hz, 1H), 7.52 (ddd, J = 8.7, 7.3, 1.7 Hz, 1H), 7.16–7.11 (m, 2H), 4.09 (s, 3H), 3.92 (s, 3H), 3.91 (s, 3H). 13 13C NMR (100 MHz, CDCl3) δ 177.6, 156.6, 156.6, 155.8, 153.8, 149.1, 137.7, 134.0, 134.0, 129.7, 121.3, 120.1, 118.0, 107.9, 99.8, 62.5, 62.3, 61.7. HRESIMS m / z 423.0075 [M + H] + (calcd for C 18 H 16 O7Br + , 423.0074);
[0032] Example 7: Compound e
[0033]
[0034] Dissolve compound a (20 mg, 1.0 eq) in chloroform, add NBS (12.4 mg, 1.2 eq), stir the reaction solution at room temperature, and monitor the reaction by TLC. After the reaction is completed, add water to terminate the reaction. Extract the reaction solution with dichloromethane three times, combine the organic phases, concentrate under reduced pressure, and separate and purify by normal-phase silica gel column chromatography and HPLC to obtain a yellow solid with a yield of 30%. 1 1H NMR (400 MHz, DMSO-d6) δ 12.40 (s, 1H), 10.44 (s, 1H), 7.62 (d, J = 2.5 Hz, 1H), 7.54 (dd, J = 8.8, 2.5 Hz, 1H), 6.97 (d, J = 8.8 Hz, 1H), 6.62 (s, 1H), 3.91 (s, 3H), 3.73 (s, 3H), 3.70 (s, 3H). 1313C NMR (100 MHz, DMSO-d6) δ 178.5, 158.3, 156.6, 155.4, 155.1, 148.6, 139.2, 134.6, 132.7, 128.3, 119.6, 118.6, 109.6, 105.0, 95.9, 61.0, 60.2, 56.6. HRESIMS m / z 423.0090 [M+H] + (calcd for C 18 H 16 O7Br + , 423.0074);
[0035] Example 8: Compound f
[0036]
[0037] Weigh compound a (30 mg, 1.0 eq) into a reaction sealed tube, add ethyl acetate to dissolve it completely, pipette DMSO (7.2 μL, 1.2 eq) and hydrobromic acid (HBr, 18.0 μL, 1.8 eq) into the reaction sealed tube. The mixed reaction solution was reacted in an oil bath at 60 °C for 2 h, and the reaction was monitored by TLC. After the reaction solution was cooled to room temperature, it was concentrated under reduced pressure and purified by normal-phase silica gel column chromatography to obtain a yellow solid with a yield of 30%. 1 1H NMR (400 MHz, CDCl3) δ 12.79 (s, 1H), 7.76 (dd, J = 7.9, 1.6 Hz, 1H), 7.67 (s, 1H), 7.65 (dd, J = 7.9, 1.6 Hz, 1H), 7.01 (t, J = 7.9 Hz, 1H), 4.08 (s, 3H), 3.92 (s, 3H), 3.90 (s, 3H). 13 13C NMR (100 MHz, CDCl3) δ 177.8, 156.6, 155.4, 153.8, 151.8, 149.0, 138.7, 136.6, 133.9, 129.6, 122.0, 119.1, 113.6, 108.0, 99.8, 62.3, 62.2, 61.7. HRESIMS m / z 500.9180 [M+H] + (calcd for C 18 H 16 O7Br2 + , 500.9179);
[0038] Example 9: Compound g
[0039]
[0040] Compound b (15 mg, 1 eq) was dissolved in dichloromethane (4 mL), followed by the addition of 1 mL of DMSO solvent to completely dissolve the compound. Subsequently, NBS (7.0 mg, 1.2 eq) was added. After reacting at room temperature for 2 h, the reaction was quenched by adding water. The mixture was extracted three times with dichloromethane solvent. The organic layers were combined and concentrated under reduced pressure to obtain the product. The product was treated by normal-phase silica gel column chromatography to obtain 7.2 mg of yellow solid with a yield of 40%. 1 H NMR (400 MHz, DMSO-d6): δ 13.03 (1H, s), 10.18 (1H, s), 7.61 (1H, dd, J = 8.0, 1.6 Hz), 7.43 (1H, dd, J = 7.7, 1.6 Hz), 7.02 (1H, ddd, J = 8.0, 7.7, 1.6 Hz), 4.01 (3H, s), 3.82 (3H, s), 3.74 (3H, s). 13 C NMR (100 MHz, DMSO-d6): δ 178.4, 155.1, 152.7 (C×2), 149.0, 140.0, 133.2, 132.5, 129.6 (CH×2), 121.7, 120.3, 119.5, 108.0, 97.7, 62.0, 61.5, 60.2. HRESIMS m / z 456.9677 [M+H] + (calcd for C 18 H 15 O7BrCl + , 456.9684).
[0041] Example 10: Antibacterial Activity Test
[0042] (1) Test Strains: Gram-positive bacteria (Staphylococcus aureus S. aureus, Methicillin-resistant Staphylococcus aureus MRSA, Staphylococcus albus S. albus, Micrococcus luteus Micrococcus luteus); Gram-negative bacteria (Pseudomonas aeruginosa Pseudomonas aeruginosa, Escherichia coli Escherichia coli).
[0043] (2) Sample Preparation: All compounds of the present invention were formulated into a stock solution of 3.2 mg / mL. They were completely dissolved and mixed using a vortex oscillator and ultrasound, and then reserved for use.
[0044] (3) Activity test: According to the CLSI antimicrobial experimental standard, the inhibitory activities of all compounds of the present invention against Gram-positive bacteria and Gram-negative bacteria were determined by the microbroth dilution method. After the test strains were activated for 2 generations on LB medium, single colonies were picked and inoculated into LB medium. After culturing at 37 °C and 200 rpm until the logarithmic growth phase, the bacterial liquid concentration was diluted to 1×10 5 CFU / mL for standby. 198 μL of the above-mentioned bacterial liquid and 2 μL of the drug solution were added to the 96-well plate respectively, so that the final concentrations were 32, 16, 8, 4, 2, 1 μg / mL respectively. Positive controls [Methicillin, Ciprofloxacin, Amoxicillin and Ampicillin] and negative controls were set, and each concentration was measured in parallel 3 times.
[0045] (4) Result observation: After the 96-well plate was cultured at 37 °C for 12 h, the OD 600 value was measured. The minimum inhibitory concentration (MIC) was defined as the concentration at which the in vitro antibacterial activity of the compound reached 80% inhibition.
[0046] Test results:
[0047] Table 1 Inhibitory activities of the compounds of the present invention against bacteria
[0048]
[0049] Note: In the table, "++++" indicates MIC ≤ 4 μg / mL; "+++" indicates 4 μg / mL < MIC ≤ 8 μg / mL; "++" indicates 8 μg / mL < MIC ≤ 16 μg / mL; "+" indicates 16 μg / mL < MIC ≤ 32 μg / mL; "-" indicates MIC > 32 μg / mL; "ns" indicates not tested.
[0050] Note: Compounds a and compound b as well as compounds c-g of the present invention were tested in parallel as controls.
[0051] Conclusion: After the activity tests on 6 kinds of bacteria, it was found that only compounds 1-4 of the present invention showed significant inhibitory activities against Staphylococcus aureus and methicillin-resistant Staphylococcus aureus, almost equivalent to the positive drugs. The same type of compounds a-g did not show inhibitory effects at the maximum test concentration. At the same time, compounds 1-4 of the present invention showed high selectivity, only showing inhibitory activities against Staphylococcus aureus and methicillin-resistant Staphylococcus aureus, and not showing activities against other tested Gram-positive bacteria and Gram-negative bacteria. Therefore, compounds 1-4 of the present invention have great application prospects in the treatment of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus.
[0052] Finally, it should be noted that there are other ways to implement the present invention; accordingly, the embodiments of the present invention are illustrated by way of example and are not limited to the content described in the present invention, and may also be modifications made within the scope of the present invention or equivalent content added in the claims; all publications or patents cited in the present invention will be used as references for the present invention.
Claims
1. A compound 1-4 of formula I structure or a pharmaceutically acceptable salt thereof or a solvate of the salt, characterized in that The compound has the following structure: wherein X1 and X2 are each independently and optionally H, Cl, Br, or I; specifically, 2. An agent against Staphylococcus aureus and methicillin-resistant Staphylococcus aureus, characterized in that Containing the compound of formula I as claimed in claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
3. Use of the compound of formula I as claimed in claim 1 or a pharmaceutically acceptable salt thereof in the preparation of an agent against Staphylococcus aureus and methicillin-resistant Staphylococcus aureus.