Application of naphthoquinone furan piperidone derivative in preparation of medicine for inhibiting bacteria

Abstract

The invention discloses an application of a naphthoquinone furan piperidone derivative in preparation of a medicine for inhibiting bacteria. The naphthoquinone furan piperidone derivative disclosed by the invention has remarkable antibacterial activity on a plurality of gram-positive bacteria. The naphthoquinone furan piperidone derivative disclosed by the invention can be used for remarkably inhibiting the growth of MRSA (Methicillin-Resistant Staphylococcus Aureus) and shows relatively low drug resistance within 30 days. The naphthoquinone furan piperidone derivative can protect MRSA infected mice and improve the survival rate of the infected mice. The naphthoquinone furan piperidone derivative disclosed by the invention shows good safety and biocompatibility while keeping high antibacterial activity.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to the application of naphthoquinone furanoperidone derivatives in the preparation of antibacterial drugs. Background Technology

[0002] The overuse of antibiotics has led to a significant increase in antimicrobial resistance (AMR), making the treatment of refractory infections increasingly difficult. Among these, methicillin-resistant Staphylococcus aureus (MRSA) is particularly prevalent. Staphylococcus aureus MRSA (metastatic antibiotics), as a representative pathogen of Gram-positive drug-resistant bacteria, has a mortality rate 64% higher than that of other bacterial infections, causing more than 100,000 direct deaths annually. However, the number of new antibiotic classes introduced in recent decades has been extremely limited, and the shortage of novel antimicrobial drugs urgently requires breakthroughs in innovative treatment strategies.

[0003] Against this backdrop, natural products (NPs) have become an important breakthrough due to their unique antibacterial mechanisms. In particular, plant-derived natural products have accumulated a high diversity of secondary metabolites during long-term co-evolution with environmental stress. Their antibacterial mechanisms have the natural advantage of not easily inducing drug resistance, which provides an opportunity to develop new drugs against drug-resistant bacteria.

[0004] Quinones are a very important class of natural products. Quinone-based derivatives possess potential pharmacological activities, such as antitumor, antibacterial, antifungal, antiviral, anti-Alzheimer's disease, and antimalarial activity. For example, patent application CN114748455A discloses the use of 5-substituted naphthoquinone compounds in the preparation of antibacterial drugs. Patent application CN103396389A discloses a class of naphthoquinone sesquiterpenes and their use in the preparation of antitumor or antibacterial drugs.

[0005] Previously, the inventors synthesized a class of naturally derived naphthoquinone-furo-piperidone compounds as dual-targeting agents of STAT3 and NQO1, exhibiting anti-breast cancer bioactivity (Shengying Lou et al., Discovery of naphthoquinone-furo-piperidone derivatives as dual-targeting agents of STAT3 and NQO1 for the treatment of breast cancer). European Journal of Medicinal Chemistry(287 (2025) 117377). However, there are no studies on the antibacterial activity of these compounds. This invention is the first to discover that these compounds have anti-MRSA activity, and they are expected to become novel compounds for treating MRSA infection, providing more options for the treatment of MRSA infection. Summary of the Invention

[0006] The purpose of this invention is to provide the use of naphthoquinone furanoperidone derivatives in the preparation of antibacterial drugs.

[0007] The application of naphthoquinone furanoperidone derivatives in the preparation of antibacterial drugs; the general structural formula of naphthoquinone furanoperidone derivatives is shown in Formula I or Formula II. Formula I, Formula II, Wherein, R is selected from substituted or unsubstituted alkyl groups and substituted or unsubstituted aryl groups.

[0008] Preferably, the structure of the naphthoquinone furanoperidone derivative is selected from any one of formulas III to VIII. , Among them, R 1 Selected from H, substituted or unsubstituted C1-C5 alkyl, substituted or unsubstituted aryl; R 2 Selected from H and CH3, n is selected from 0 to 2; The connector in formula VI is selected from substituted or unsubstituted C3-C. 10 Cycloalkyl, substituted or unsubstituted C1-C3 alkyl carbonyloxy, substituted or unsubstituted C6-C 12 Aryl group, element X is selected from C or O; The terminal bases of Formulas VII and VIII are selected from dimethylamino, diethylamino, piperidinyl, substituted or unsubstituted morpholino, substituted or unsubstituted piperazine, imidazole, or spirocyclic groups containing heteroatoms, and n is selected from 1 to 2.

[0009] Preferably, the structure of the naphthoquinone furanoperidone derivative is selected from: , .

[0010] Preferably, the bacteria include Gram-positive bacteria.

[0011] More preferably, the Gram-positive bacteria include at least one of Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, Staphylococcus epidermidis, and Streptococcus pyogenes.

[0012] More preferably, the Staphylococcus aureus is at least one of methicillin-resistant Staphylococcus aureus and quinolone-resistant Staphylococcus aureus.

[0013] Preferably, the naphthoquinone furanoperidone derivative is the naphthoquinone furanoperidone derivative itself or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs.

[0014] Preferably, the inhibiting bacteria are bacteria that inhibit bacteria in the environment or bacteria in the body of animals that inhibit bacterial infections.

[0015] The present invention has the following advantages and beneficial effects: The naphthoquinone furanopiperidone derivatives of the present invention exhibit significant antibacterial activity against a variety of Gram-positive bacteria. These derivatives significantly inhibit the growth of MRSA and show low resistance within 30 days. They also protect MRSA-infected mice and improve their survival rate. Furthermore, while maintaining high antibacterial activity, these derivatives demonstrate good safety and biocompatibility. Attached Figure Description

[0016] Figure 1 The time-growth curve of MRSA after Y-22 treatment in Example 2 is shown.

[0017] Figure 2 The image shows the bactericidal kinetic curve of Y-22 against MRSA in Example 3.

[0018] Figure 3 The inhibition zone of Y-22 against MRSA in Example 4.

[0019] Figure 4 This is a graph showing the results of the 30-day drug resistance analysis of Y-22 in Example 5.

[0020] Figure 5 This refers to the inhibitory activity of Y-22 on MRSA biofilm formation in Example 6.

[0021] Figure 6 This refers to the hemolytic activity of Y-22 in Example 7.

[0022] Figure 7 The survival curves of MRSA-infected peritonitis mice in Example 8 are shown. Detailed Implementation

[0023] The present invention adopts the following technical solution: The present invention provides the use of a compound of formula (I) or formula (II) or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof in the preparation of a medicament for inhibiting methicillin-resistant Staphylococcus aureus.

[0024] Formula I, Formula II.

[0025] in, R is selected from substituted or unsubstituted alkyl groups, and substituted or unsubstituted aryl groups.

[0026] Furthermore, the structure can be selected from Formula III to Formula VIII.

[0027] , Furthermore, R in Equations III to V 1 Selected from H, substituted or unsubstituted C1-C5 alkyl, substituted or unsubstituted aryl, R 2 Selected from H, CH3, n selected from 0 to 2; the linker in formula VI is selected from substituted or unsubstituted C3-C 10 Cycloalkyl, substituted or unsubstituted C1-C3 alkyl carbonyloxy, substituted or unsubstituted C6-C 12 The aryl group, element X is selected from C and O; the terminal bases of formulas VII and VIII are selected from dimethylamino, diethylamino, piperidinyl, substituted or unsubstituted morpholino, substituted or unsubstituted piperazine, imidazole, spirocyclic group containing heteroatoms (such as N and O), and n is selected from 1 to 2.

[0028] Furthermore, the compounds represented by formulas (I) and (II) are selected from: , .

[0029] Unless otherwise stated, the term "alkyl" as used herein includes branched and straight-chain saturated aliphatic hydrocarbon groups having a specific number of carbon atoms, including all isomers.

[0030] The term "carbonyl" refers to an organic functional group (C=O) formed by carbon and oxygen atoms linked by a double bond.

[0031] The term "cycloalkyl" refers to a hydrocarbon group in a monocyclic system of carbon atoms with a saturated ring.

[0032] The term "aryl" refers to aromatic single- or multi-carbon ring systems. In a multi-carbon ring system, the carbon rings are fused together or connected to each other by single bonds. In the case of multi-carbon rings, as long as one carbon ring is an aromatic ring, it is acceptable. Generally, aryl groups include phenyl, naphthyl, and biphenylene.

[0033] In this invention, "C1-C8 alkyl" refers to a straight-chain or branched alkyl group having a specific number of carbon atoms (1-8), including all isomers, including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, etc. "C2-C8 alkenyl" refers to a straight-chain or branched alkyl group having a specific number of carbon atoms (2-8), including all isomers, including but not limited to vinyl, allyl, etc. "C3-C8 cycloalkyl or cycloalkenyl" refers to a hydrocarbon group with a 3-8 carbon atom monocyclic system having a saturated or unsaturated ring, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopropenyl, cyclohexenyl, etc. Similarly, the term "C1-C8 alkyl" is used in conjunction with "C1-C8 alkyl" in this invention. 10 Alkyl group, C2-C 10 "Alkenyl", "C1-C8 carbonyl", "C3-C8 cycloalkenyl", "C6-C 10 Aryl", C5-C 10 Heterocyclic group, C5-C 10 "Mild aryl" and similar terms have similar meanings.

[0034] Unless otherwise stated, all ranges listed in this article are inclusive. For example, "1-4" includes 1, 2, 3, and 4.

[0035] The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic alkali or acid. When the compounds of the present invention are acidic, their corresponding salts can be readily prepared from inorganic or organic bases. Salts derived from such inorganic bases include aluminum, ammonium, calcium, copper (copper and cuprous), iron, ferrous, lithium, magnesium, manganese (manganese and manganese), potassium, sodium, zinc, etc. Preferred are ammonium, calcium, magnesium, potassium, and sodium salts. Salts prepared from organic bases include primary, secondary, and tertiary amines derived from natural and synthetic sources. Pharmaceutically acceptable non-toxic organic bases that can form salts include arginine, betaine, caffeine, choline, etc. N,N′ -Dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N -Ethylmorpholine, N-Ethylpiperidine, glucosamine, glucosamine, histidine, hydrabamine, isopropylamine, dicyclohexylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, polyamine resin, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. When the compounds of the present invention are basic, their corresponding salts can be readily prepared from inorganic or organic acids. Such acids include, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, hydroxyethylsulfonic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, viscous acid, nitric acid, pyric acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, etc. In this invention, the preferred salts are those formed with hydrochloric acid, nitric acid, sulfuric acid, phosphoric acid, hydrobromic acid, maleic acid, fumaric acid, tartaric acid, lactic acid, citric acid, acetic acid, methanesulfonic acid, p-toluenesulfonic acid, adipic acid, palmitic acid, tannic acid, lithium, sodium, potassium, calcium, magnesium, and lysine.

[0036] The term "solvate" refers to a complex of variable stoichiometry formed by a solute (i.e., compounds of formula I and II) or a pharmaceutically acceptable salt thereof and a solvent that does not interfere with the biological activity of the solute. Examples of solvents include, but are not limited to, water, ethanol, and acetic acid. When the solvent is water, the solvate is called a hydrate. Hydrates include, but are not limited to, hemihydrates, monohydrates, sesquihydrates, dihydrates, and trihydrates.

[0037] The term "prodrug" refers to a functional derivative of the compound of this invention, which is readily converted into the desired compound in vivo. In this invention, various derivatives obtained by reducing quinone to diphenol, and prodrug derivatives with other substituents are preferred.

[0038] The naphthoquinone-furo-piperidone derivative of this invention is obtained by chemical synthesis. The synthetic method is detailed in the inventors' previously published article (Shengying Lou et al., Discovery of naphthoquinone-furo-piperidone derivatives as dual targeting agents of STAT3 and NQO1 for the treatment of breast cancer). European Journal of Medicinal Chemistry287 (2025) 117377). In vitro antibacterial experiments, cytotoxicity experiments, hemolytic experiments, drug resistance experiments, and bacterial biofilm formation experiments showed that these naphthoquinone furapiridone derivatives possess broad-spectrum antibacterial activity against Gram-positive bacteria, good biocompatibility, low drug resistance, and the ability to inhibit bacterial biofilm formation. Animal experiments showed that naphthoquinone furapiridone derivatives have good therapeutic and protective effects against MRSA-infected mice and improve the survival rate of infected mice. Naphthoquinone furapiridone derivatives hold promise for enriching the antibacterial drug library, especially in the area of ​​methicillin-resistant Staphylococcus aureus, where they have significant potential for discovery.

[0039] This invention has discovered that naphthoquinone furanoperidone derivatives exhibit significant inhibitory effects on MRSA and good safety, and also provide some protection against MRSA-infected mice. Based on this, this invention provides the use of naphthoquinone furanoperidone derivatives in any of the following: (1) Application of naphthoquinone furanoperidone derivatives in antibacterial applications.

[0040] (2) Application of naphthoquinone furanoperidone derivatives in the preparation of antibacterial agents and preparations for the prevention of bacterial infection.

[0041] (3) Application of naphthoquinone furanoperidone derivatives in the preparation of drugs and their formulations that improve the survival rate of bacterial infection receptors.

[0042] Optionally, the bacteria include Gram-positive bacteria.

[0043] Preferably, the Gram-positive bacteria include Staphylococcus aureus, methicillin-resistant Staphylococcus aureus, quinolone-resistant Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, Staphylococcus epidermidis, and Streptococcus pyogenes.

[0044] Optionally, the receptor may include an animal.

[0045] The present invention also provides an anti-Gram-positive bacteria drug comprising a naphthoquinone furanoperidone derivative and pharmaceutically acceptable excipients and carriers.

[0046] Optionally, the Gram-positive bacteria include Staphylococcus aureus, methicillin-resistant Staphylococcus aureus, quinolone-resistant Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, Staphylococcus epidermidis, and Streptococcus pyogenes, etc.

[0047] Example 1: Determination of the minimum inhibitory concentration of naphthoquinone furanoperidolone derivatives against Gram-positive bacteria According to the CLSI antimicrobial susceptibility testing standard, the minimum inhibitory concentration (MIC) of naphthoquinone furapiridone derivatives against MRSA was determined using the serial two-fold dilution method. Ciprofloxacin and vancomycin were used as positive controls. The 96-well plates were incubated at 37°C, and the lowest drug concentration at which no bacterial growth was observed visually was defined as the MIC value of the naphthoquinone furapiridone derivative against bacteria.

[0048] The results are shown in Table 1. The MIC values ​​of naphthoquinone furanoperidone derivatives for MRSA were generally between 4 and 32 μg / mL.

[0049] Table 1. MIC values ​​of naphthoquinone furanoperidone derivatives for MRSA

[0050] Y-22, a representative compound with good antibacterial activity, was selected for further research. The antibacterial activity of Y-22 against various Gram-positive bacteria was tested. The results are shown in Table 2. The MIC values ​​of Y-22 against various Gram-positive bacteria ranged from 2 to 64 μg / mL.

[0051] Table 2 MIC values ​​of Y-22 against various Gram-positive bacteria

[0052] Example 2: Time-growth curve of Y-22 on MRSA After bacterial activation, the bacteria were diluted with TSB medium and six groups were set up (blank control group, 1 / 16MIC, 1 / 8MIC, 1 / 4MIC, 1 / 2MIC, and MIC), and the compound was added for treatment. At each time point (0h, 2h, 4h, 6h, 8h, 10h, 12h, and 24h), 100 μL of bacterial solution was taken from the centrifuge tube and transferred to a 96-well plate. The remaining solution was placed in a shaker at 37°C for further incubation, and the absorbance at 600 nm was measured.

[0053] The results are as follows Figure 1 As shown in the figure. Compared with the blank control group, Y-22 effectively inhibited the growth of MRSA at 1 / 2 MIC.

[0054] Example 3: Time-sterilization curve of Y-22 against MRSA After bacterial activation, the culture was diluted with TSB medium, and five groups were set up (MIC, 2MIC, 4MIC, 8MIC, and a blank control group). Compound was added for each group. At each time point (0h, 2h, 4h, 6h, 8h, 10h, 12h, and 24h), 100 μL of bacterial culture was taken from a centrifuge tube, and the remainder was incubated at 37°C in a shaker. The extracted bacterial culture was then diluted tenfold and plated. After overnight incubation, the colonies were counted, and the number of colonies per milliliter was calculated.

[0055] The results are as follows Figure 2 As shown in the figure. Compared with the blank control group, Y-22 showed a significant bactericidal effect at concentrations of 2×MIC and above.

[0056] Example 4: Antibacterial zone experiment Punch small circular pieces of filter paper using a punch and autoclave. Spread the diluted bacterial suspension onto TSA plates and allow it to sit for an appropriate time. Place four filter papers on the plate and add the prepared concentrations of the compound (Y-22 concentrations of 1.6 μg, 3.2 μg, and 6.4 μg). Incubate at 37°C for 14–16 h.

[0057] The results are as follows Figure 3 As shown in the figure. Compared with the blank control group, Y-22 formed a clear inhibition zone around the plate after administration, demonstrating good antibacterial activity.

[0058] Example 5: Drug Resistance Test After MRSA activation, the bacterial count was adjusted by dilution. Y-22 was co-cultured with the bacteria at a concentration of 1 / 2 MIC. After 24 hours of culture, the MIC was continuously measured over 30 days in the same manner, and a curve was plotted with the change in MIC as the ordinate.

[0059] The results are as follows Figure 4 As shown, Y-22 exhibited lower induced resistance characteristics within 30 days compared to ciprofloxacin (CIP).

[0060] Example 6: Bacterial Biofilm Formation Experiment MRSA and Y-22 were co-cultured. After culture, the culture medium was aspirated with a pipette, and the cells were washed once with PBS buffer. 99% methanol solution was added to each well for fixation. After fixation, crystal violet staining solution was added for 20 min, followed by rinsing with tap water to remove unadsorbed crystal violet staining solution, and the cells were air-dried at room temperature. 33% glacial acetic acid solution was added to each well, and the absorbance at 570 nm was measured using a microplate reader.

[0061] The results are as follows Figure 5As shown, Y-22 treatment inhibited biofilm formation in MRSA, with a significant difference between the 1 / 4 MIC group and the blank control group (Control group) with added DMSO.

[0062] Example 7: Hemolytic Activity Test 1% mouse red blood cells (mRBCs) were centrifuged and resuspended to obtain a 5% (v / v) suspension. The compound was added for treatment, with a blank control group (no red blood cells added, only PBS added; the PBS group is shown in the figure), a 1% Triton X-100 positive control group (Triton X group is shown in the figure), and a DMSO negative control group (Control group is shown in the figure). The mixture was incubated at 37°C for 1 h, centrifuged, and photographed. 100 μL of the supernatant was transferred to a 96-well plate, and the absorbance was measured at 450 nm using a microplate reader.

[0063] The results are as follows Figure 6 As shown, the hemolysis rate of Y-22 is less than 10% at concentrations below 512 μg / mL, indicating good biocompatibility.

[0064] Example 8: MRSA-infected mouse peritonitis experiment MRSA activation, followed by counting, and then prepared into 5×10⁻⁶ solutions. 8 The bacterial suspension at CFU / mL was administered intraperitoneally to each mouse, with 0.2 mL injected into each mouse. Two hours later, the mice were administered the following intraperitoneal injections: saline group (NS group), model group (vehicle group), positive control group (vancomycin (VAN) 10 mg / kg), low-dose (15 mg / kg) and high-dose (30 mg / kg) Y-22 groups. The survival rate of mice was observed within 48 hours.

[0065] The results are as follows Figure 7 As shown, Y-22 showed a protective effect against infected mice at low doses, with a survival rate of 33.3%. This protective effect was more pronounced at high doses, with the survival rate increasing to 50%.

Claims

1. The application of naphthoquinone furanoperidone derivatives in the preparation of antibacterial drugs, characterized in that, The general structural formulas of naphthoquinone furanoperidinone derivatives are shown in Formula I or Formula II. Formula I, Formula II, Wherein, R is selected from substituted or unsubstituted alkyl groups and substituted or unsubstituted aryl groups.

2. The application according to claim 1, characterized in that, The structures of naphthoquinone furanoperidone derivatives are selected from any one of formulas III to VIII. ， Among them, R 1 Selected from H, substituted or unsubstituted C1-C5 alkyl, substituted or unsubstituted aryl; R 2 Selected from H and CH3, n is selected from 0 to 2; The connector in formula VI is selected from substituted or unsubstituted C3-C. 10 Cycloalkyl, substituted or unsubstituted C1-C3 alkyl carbonyloxy, substituted or unsubstituted C6-C 12 Aryl group, element X is selected from C or O; The terminal bases of Formulas VII and VIII are selected from dimethylamino, diethylamino, piperidinyl, substituted or unsubstituted morpholino, substituted or unsubstituted piperazine, imidazole, or spirocyclic groups containing heteroatoms, and n is selected from 1 to 2.

3. The application according to claim 1, characterized in that, The structures of naphthoquinone furanoperidone derivatives are selected from: ， 。 4. The application according to claim 1, characterized in that, The bacteria include Gram-positive bacteria.

5. The application according to claim 4, characterized in that, The Gram-positive bacteria include at least one of Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, Staphylococcus epidermidis, and Streptococcus pyogenes.

6. The application according to claim 5, characterized in that, The Staphylococcus aureus is at least one of methicillin-resistant Staphylococcus aureus and quinolone-resistant Staphylococcus aureus.

7. The application according to claim 1, characterized in that, Naphthoquinone furanoperidone derivatives are naphthoquinone furanoperidone derivatives themselves or their stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs.

8. The application according to claim 1, characterized in that, Inhibit bacteria refers to the inhibition of bacteria in the environment or the inhibition of bacterial infections in animals.

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