Application of anhydride compound in preparation of bactericide drugs
By preparing acid anhydride compounds 1 and 2 as fungicides, the problem of drug resistance of citrus pathogens and human pathogens was solved, and the development of a fungicide with high inhibitory effect and environmental friendliness was achieved.
Patent Information
- Application Number
- CN202511673111.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-14
- Publication Date
- 2026-01-13
Smart Images
Figure CN121313631A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the application of microbial secondary metabolites, specifically to the use of an acid anhydride compound in the preparation of a bactericide (Penicillium fingernail, a citrus pathogen). Penicillium digitatum ) and human pathogen Pseudomonas aeruginosa ( Pseudomonas aeruginosa Application in drugs of class )) Background Technology
[0002] In the agricultural field, Penicillium fingerlings ( Penicillium digitatum Penicillium fingering is a common pathogenic fungus affecting citrus crops and is one of the most intractable and destructive citrus diseases globally. Citrus is susceptible to Penicillium fingering. Penicillium digitatum The disease caused by infection with *Pseudomonas aeruginosa* is commonly known as citrus green mold, accounting for approximately 60% to 90% of postharvest diseases in citrus. This makes citrus fruits highly susceptible to infection during transportation and storage after harvest, resulting in severe economic losses. Pseudomonas aeruginosa *Cladosporium praecoxibaris* is a common opportunistic pathogen that can cause various infectious diseases, especially vulnerable hospitalized patients with weakened or compromised immune systems. On the other hand, due to the long-term and extensive use of bactericides and antibiotics, the drug resistance of pathogens is increasing year by year, urgently requiring the development of novel antibacterial drugs. The anhydride compounds involved in this invention utilize *Cladosporium praecoxibaris* (…). Talaromyces trachyspermus The acid anhydride compounds 1 (glaucanic acid) and 2 (glauconic acid) involved in this invention can be obtained from various microorganisms through fermentation, separation, and purification (e.g., ...). Natural Product Research , 2024, 38(4), 696; 安 Anhui Agricultural Science , 2015, 43(21), 1), but its efficacy against the citrus pathogen Penicillium finger ( Penicillium digitatum ) and human pathogen Pseudomonas aeruginosa ( Pseudomonas aeruginosa Reports on inhibitory activity. Summary of the Invention
[0003] The purpose of this invention is to provide an anhydride compound for the preparation of a bactericide (Penicillium finger-shaped fungicide for citrus pathogens). Penicillium digitatum ) and human pathogen Pseudomonas aeruginosa ( Pseudomonas aeruginosa Application in drugs of class ))
[0004] To achieve the above objectives, the technical solution adopted by the present invention is as follows: Application of an acid anhydride compound in the preparation of bactericidal drugs.
[0005] The anhydride compounds are the molecular structures of anhydride compound 1 (glaucanic acid) and compound 2 (glauconicacid), as shown in Formula I.
[0006]
[0007] The acid anhydride compound 1 (glaucanic acid) and compound 2 (glauconic acid) are used as bactericides or as lead compounds for bactericides.
[0008] The acid anhydride compounds 1 (glaucanic acid) and 2 (glauconic acid) in Formula I, as fungicides or their lead compounds, are used to control the citrus pathogen Penicillium fingernail. Penicillium digitatum and / or human pathogens Pseudomonas aeruginosa Pseudomonas aeruginosa Applications in [the context of the text].
[0009] The fungicide contains excipients available for use in biological pesticides and is prepared in various formulations, including liquid and solid formulations.
[0010] The dosage forms include powders, emulsions, aqueous solutions, granules, suspensions, water-in-oil emulsions, and suspension emulsions.
[0011] Advantages of this invention: 1. The acid anhydride compound 1 (glaucanic acid) and compound 2 (glauconicacid) involved in this invention (as shown in structural formula I) can be used as fungicides or drug lead compounds for the citrus pathogen Penicillium finger and the human pathogen Pseudomonas aeruginosa.
[0012] 2. The acid anhydride compounds involved in this invention can be produced by fermentation using microorganisms, and have the advantages of mild fermentation conditions, rapid and efficient separation and purification process, high purity of the obtained monomer compounds, easy large-scale preparation and environmental friendliness. 3. This invention utilizes microbial fermentation to prepare antibacterial compounds, which is environmentally friendly; 4. The glaucanic acid and glauconicacid compounds involved in this invention exhibit significant inhibitory activity against the growth of *Penicillium digitatum* and *Pseudomonas aeruginosa*, and can be developed into novel fungicides or as active lead compounds for novel fungicides. Experiments have shown that the MIC values of compounds 1 and 2 against *Penicillium digitatum* are 2.0 μg / mL and 1.0 μg / mL, respectively, equivalent to or better than the activity of the positive control amphotericin B (MIC 2.0 μg / mL). Furthermore, the MIC values of compounds 1 and 2 against *Pseudomonas aeruginosa* are both 2.0 μg / mL, comparable to chloramphenicol (MIC 2.0 μg / mL), and can be used as preventative agents or lead compounds against citrus pathogens and human pathogens. Detailed Implementation
[0013] The following examples further illustrate specific embodiments of the present invention. It should be noted that the specific embodiments described herein are merely for illustration and explanation and are not intended to limit the scope of the present invention.
[0014] Example 1: Application of acid anhydride compounds 1 and 2 as drugs for the prevention and control of citrus pathogen Penicillium finger and human pathogen Pseudomonas aeruginosa.
[0015] The acid anhydride compounds are compound 1 (glaucanic acid) and compound 2 (glauconic acid), as shown in Formula I:
[0016] The preparation process of anhydride compound 1 (glaucanic acid) can be found in the literature. Natural Product Research The preparation process of compound 2 (glauconic acid) is described in the literature, 2024, 38(4), 696. Anhui Agricultural Science The method is described in , 2015, 43(21), 1. It can also be synthesized using existing chemical synthesis methods.
[0017] Compound 1 and Compound 2 have the following physicochemical and spectroscopic properties: Compound 1: Colorless crystals; melting point 172-174℃; chemical formula C 18 H 20 O6; Specific rotation [α] = +185.0 ( c0.15, MeOH); UV (MeOH) λmax (log ε) 214 (0.15), 259 (–0.10) nm; ECD λmax(Δε) 207 (–2.12), 242 (2.22) nm; 1 H-NMR and 13 C-NMR data are shown in Table 1.
[0018] Compound 2: Colorless crystals; melting point 175-177℃; chemical formula C 18 H 20 O7; Specific rotation [α] = +33.0 ( c 1.02, MeOH); UV (MeOH) λmax (log ε) 216 (1.82), 261 (0.13) nm; ECD λmax (Δε) 215 (–51.66), 242 (30.12), 279 (9.28) nm; 1 H-NMR and 13 C-NMR data are shown in Table 1.
[0019] Table 1. 1H NMR (500 MHz) and 1C NMR (125 MHz) data for compounds 1 and 2
[0020] a Deuterated solvent CDCl3; b Deuterated solvent DMSO- d 6.
[0021] Example 2: Inhibitory activity against agricultural pathogens and human pathogens The inhibitory activity of compounds 1 and 2 (shown in Formula I) against agricultural pathogens and human pathogens was determined using the minimum inhibitory concentration (MIC) method. The following agricultural pathogens and human pathogens were selected as test bacteria: Agricultural pathogens: Penicillium fingerlings Penicillium digitatum Human pathogen: Pseudomonas aeruginosa Pseudomonas aeruginosa Antibacterial activity test: 1) Minimum inhibitory concentration (MIC) assay: The minimum inhibitory concentration (MIC) is the lowest concentration of a drug that can inhibit the growth of microorganisms in vitro. In a 96-well microplate, different concentrations of the drug are added to the bacterial suspension of the test bacteria. After incubation, observation is performed. If the indicator bacteria can grow in a well, it means that the corresponding drug concentration in that well cannot inhibit the growth of the bacteria; the liquid in that well will be turbid, and the light transmittance will be significantly reduced. Conversely, if the indicator bacteria cannot grow in a well, the liquid in that well will be clear, and the decrease in light transmittance will not be significant. The lowest sample concentration that completely inhibits the growth of the indicator bacteria in a well is the minimum inhibitory concentration (MIC) of the compound.
[0022] 2) Preparation of bacterial suspension: The tested bacteria were inoculated into culture media (Sabella medusae var. agar for Penicillium fingerlings and LB broth for Pseudomonas aeruginosa) and incubated at 28°C for 24 hours. Afterward, 4 mL of sterile 0.85% NaCl solution (fungal physiological saline containing 0.25% Tween 20) was used to wash the culture medium, and the hyphae were gently scraped off with a glass scraper. An appropriate amount of the hyphal suspension was pipetted into a sterile test tube, and then adjusted to 0.5 McFarland turbidity (equivalent to 1.5 × 10⁻⁶) with 0.85% NaCl solution. 8 The concentration was (CFU / mL) and further diluted to 5 × 10⁻⁶ with 0.85% NaCl solution. 5 CFU / mL.
[0023] 0.5 McFarland turbidity standard: Add 0.5 mL × 0.048 mol / L BaCl2 (1.175% w / v BaCl2·2H2O) to 99.5 mL × 0.18 mol / L (0.36 N) H2SO4 (1% v / v) and stir continuously to maintain suspension.
[0024] 3) Sample preparation: Take approximately 1 mg of the test samples (compounds 1 and 2 obtained above) and positive controls (amphotericidal B and chloramphenicol), respectively, and dissolve them in approximately 100 μL of DMSO. After thorough mixing, the final concentration is 2560 μg / mL. Pipette 50 μL of the sample solution into another centrifuge tube, and then add 50 μL of DMSO to obtain a sample solution with half the concentration. Repeat this process sequentially to obtain a total of 11 sets of sample solutions with successively halved concentrations (serial dilutions) (2560, 1280, 640, 320, 160, 80, 40, 20, 10, 5, 2.5 μg / mL).
[0025] 4) Blank control: Select the pure solvent that has not dissolved the test sample as a blank control.
[0026] 5) MIC measurement procedure: 5.1) Using aseptic technique, the sample solutions of different concentrations after serial dilution were added to sterile 96-well plates. 5 μL of sample solution was added to wells 1 to 11, and well 12 was left untreated as a growth control.
[0027] 5.2) Dilute the indicator bacterial suspension (equivalent to 0.5 McFarland turbidity) 1000 times with liquid culture medium (Sabourellow's medium for Penicillium digitatum, LB medium for Pseudomonas aeruginosa), and add 95 μL of each solution sequentially to 96-well plates, so that the final sample concentrations in wells 1 to 11 are 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, and 0.125 μg / mL, respectively. Gently shake to mix, seal the 96-well plate and incubate at 28°C for 24 h before observation.
[0028] 5.3) Measure the absorbance of each well at 600 nm using a microplate reader. The lowest sample concentration that completely inhibits the growth of indicator bacteria in the well is taken as the MIC value of the compound. Note: The experiment is only meaningful when the indicator bacteria show obvious growth in the growth control well; when a single well skips a test, the highest drug concentration that inhibits the growth of the strain should be recorded; if multiple wells skip a test, the results should not be reported, and the experiment should be repeated.
[0029] The experimental results showed that compounds 1 and 2 were effective against the citrus pathogen Penicillium fingernail. Penicillium digitatum Both compounds exhibited significant inhibitory activity, with minimum inhibitory concentrations (MICs) of 2.0 μg / mL and 1.0 μg / mL, respectively. The MIC of the positive control amphotericin B was 2.0 μg / mL. Compounds 1 and 2 also showed significant inhibitory activity against the human pathogen *Pseudomonas aeruginosa*. Pseudomonas aeruginosa The minimum inhibitory concentration (MIC) was 2.0 μg / mL, which was comparable to that of the positive control chloramphenicol (MIC = 2.0 μg / mL).
[0030] The above experimental results demonstrate that the compounds involved in this invention have a strong inhibitory effect on agricultural pathogens such as Penicillium finger and human pathogens such as Pseudomonas aeruginosa, and can be used to prepare novel fungicides or as lead compounds for fungicides.
Claims
1. The application of an acid anhydride compound in the preparation of bactericidal drugs.
2. The application according to claim 1, characterized in that: The anhydride compounds are the molecular structures of anhydride compound 1 (glaucanic acid) and compound 2 (glauconic acid), as shown in Formula I: 。 3. The application according to claim 2, characterized in that: The use of the acid anhydride compound 1 (glaucanic acid) and compound 2 (glauconic acid) as bactericides or lead compounds for bactericides.
4. The application according to claim 3, characterized in that: The acid anhydride compounds 1 (glaucanicacid) and 2 (glauconic acid) in Formula I, as fungicides or their lead compounds, are used to control the agricultural pathogen Penicillium fingernail. Penicillium digitatum and / or human pathogens Pseudomonas aeruginosa Pseudomonas aeruginosa Applications in [the field].