Application of fully substituted 1,2,3-triazole-4-phosphonate compounds in the preparation of antibacterial drugs

Abstract

The present invention discloses the application of fully substituted 1,2,3-triazole-4-phosphonate compounds in the preparation of antibacterial drugs, especially the application of fully substituted 1,2,3-triazole-4-phosphonate compounds in the preparation of antibacterial drugs for plant pathogenic bacteria Rhizoctonia solani. The present invention studies the biological activities of different products and finds that three compounds with specific structures selectively have a strong inhibitory effect on the growth of plant pathogenic bacteria Rhizoctonia solani, and can be further used to prepare antibacterial drugs for plant pathogenic bacteria Rhizoctonia solani.

Description

Technical Field

[0001] The present invention relates to a new medical application of fully substituted 1,2,3-triazole-4-phosphonate compounds, and specifically relates to the application of fully substituted 1,2,3-triazole-4-phosphonate compounds in the preparation of antibacterial drugs. Background Art

[0002] Triazoles are a very important class of heterocyclic compounds. They have a wide range of applications in related fields such as materials chemistry, medicinal chemistry, organic synthesis, biomolecular synthesis, and biological activity. The synthesis of triazoles through the click reaction can easily assemble a pair of complementary tetrahedral building blocks into porous organic polymers (POPs). POP materials connected by triazole heterocycles have good thermal and chemical stability and can be used as excellent engineering materials under a variety of conditions. Click reactions occur between azide-functionalized metal organic frameworks UiO-66-N3 and DNA attached with dibenzylcyclooctyne. The surface of MOF is covalently functionalized with oligonucleotides through triazole heterocycles to synthesize nucleic acid-MOF nanoparticle conjugates. When dispersed in NaCl aqueous solution, they have higher stability and stronger cellular uptake ability than unfunctionalized MOF particles of the same size. In addition, nucleoside analogs with 1,4,5-persubstituted 1,2,3-triazolyl-glycosides have effective anti-leukemia effects in several hematopoietic cell lines.

[0003] Phosphorus-substituted triazole compounds, especially phosphate-substituted and phosphate-substituted, are important and stable phosphothistidine analogs for studying protein phosphorylation. Phosphohistidine is extremely unstable because the phosphate group is connected to the N atom on the imidazole ring to form a phosphoramide structure, its PN chemical bond is easily hydrolyzed, and it is sensitive to acid. This has caused long-term technical difficulties in its research, so the demand for related analogs is particularly urgent. The Muir team successfully designed an analog of phosphothistidine by converting the easily hydrolyzed PN bond into a difficult-to-decompose CP covalent bond and transforming the diazole ring into a triazole ring, and has been used in solid-phase peptide synthesis and protein semi-synthesis.

[0004]

[0005] Fully substituted phosphorus-triazole compounds have attracted widespread attention because they have more functional groups and are more diverse than disubstituted ones. At present, there is no record of fully substituted 1,2,3-triazole-4-phosphonate compounds being used to prepare antibacterial drugs against the plant pathogen Rhizoctonia solani. Summary of the invention

[0006] The purpose of the present invention is to provide a new medical application of fully substituted 1,2,3-triazole-4-phosphonate compounds.

[0007] The present invention adopts the following technical solution to solve the above technical problems: application of fully substituted 1,2,3-triazole-4-phosphonate compounds in the preparation of antibacterial drugs, characterized in that the structural formula of the fully substituted 1,2,3-triazole-4-phosphonate compounds is:

[0008] It is further defined that the fully substituted 1,2,3-triazole-4-phosphonate compound is used in the preparation of antibacterial drugs against plant pathogens.

[0009] It is further specified that the plant pathogen is Rhizoctonia solani.

[0010] It is further defined that the concentration of the fully substituted 1,2,3-triazole-4-phosphonate compound in the bacterial solution is 0.5 to 4 mg / mL.

[0011] Compared with the prior art, the present invention has the following advantages and beneficial effects: Through the study of the biological activities of different products, the present invention found that three compounds with specific structures selectively have a strong inhibitory effect on the growth of the plant pathogen - Rhizoctonia solani, and can be further used to prepare antibacterial drugs against the plant pathogen - Rhizoctonia solani. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 This is a graph showing the antibacterial effects of different compounds on Escherichia coli;

[0013] Figure 2 This is a graph showing the antibacterial effects of different compounds on Staphylococcus aureus;

[0014] Figure 3 This is a graph showing the antibacterial effects of different compounds on yeast X33;

[0015] Figure 4 This is the effect diagram of the antibacterial experiment of different compounds on yeast YJM789;

[0016] Figure 5 This is the antibacterial effect diagram of product A on plant pathogenic bacteria-Rhizoctonia solani;

[0017] Figure 6 This is the antibacterial effect diagram of product B on plant pathogenic bacteria-Rhizoctonia solani;

[0018] Figure 7 This is the antibacterial effect of product C on plant pathogenic bacteria-Rhizoctonia solani

[0019] Figure 8 This is a comparison chart of the antibacterial effects of different compounds on the plant pathogen Rhizoctonia solani. DETAILED DESCRIPTION

[0020] The above contents of the present invention are further described in detail below through examples, but this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples, and all technologies implemented based on the above contents of the present invention belong to the scope of the present invention.

[0021] Example 1

[0022] Synthesis of fully substituted 1,2,3-triazole-4-phosphonate compounds:

[0023]

[0024] Take the ionic liquid [Choline][Ac] (0.1mmol) and add it to a 10mL eggplant-shaped flask, then add the dimethyl acetonephosphonate compound 2 (0.1mmol), stir and mix evenly, and finally add the 4-trifluoromethylbenzene azide compound 1 (0.15mmol); put the flask into a 30°C oil bath to react for 24 hours; after the reaction, extract with deionized water and ethyl acetate three times, dry the organic phase with anhydrous sodium sulfate, remove the ethyl acetate with a rotary evaporator, and separate the clean product 3 by silica gel column chromatography. The different reaction substrates and synthetic products are shown in Table 1.

[0025] Table 1 Different reaction substrates and synthetic products

[0026]

[0027]

[0028] Example 2

[0029] Study on the biological activity of fully substituted 1,2,3-triazole-4-phosphonate compounds:

[0030] 1. Preparation of culture medium

[0031] LB medium: weigh 10 g of sodium chloride (NaCl), 10 g of peptone (Typtone), 5 g of yeast extract (Yeast Extract) and 15 g of agar powder (Agar, solid medium) and add them into a 1 L conical flask. Then add ddH2O to 1 L and sterilize at 115°C for 25 minutes.

[0032] YPD medium: weigh 20 g of peptone, 20 g of glucose (Dextrose), 10 g of yeast extract and agar powder (solid medium) and add them into a 1 L conical flask. Then add ddH2O to 1 L and sterilize at 115°C for 25 minutes.

[0033] PDA medium: 1L of 20wt% potato juice (200g of peeled potatoes were weighed, cut into small pieces, added with 1000mL of water and boiled for 20 minutes, the potato pieces were filtered out, and the filtrate was supplemented to 1000mL), and 20g of glucose, 3g of potassium dihydrogen phosphate (KH2PO4), 1.5g of magnesium sulfate heptahydrate (MgSO4.7H2O), 8mg of Vitanib B1 (thiamine) Trace, and 20g of agar powder (solid medium) were weighed and added to the above potato juice. Then sterilized at 115°C for 25 minutes.

[0034] After sterilization in the clean bench, unseal the sterilized culture medium in the clean bench, pour it into the sterilized culture dish, and freeze it for later use.

[0035] 2. Activation and cultivation of bacteria

[0036] Activation: Sterilize the clean bench with ultraviolet light and ethanol before the experiment, and use 75% ethanol by volume to sterilize from the hands to the elbows. Light the alcohol lamp, and try to perform all operations within 10 cm of the alcohol lamp. Use a bacterial loop (sterilize with the outer flame of the alcohol lamp before and after each use) to streak Escherichia coli, yeast X33, yeast YJM789, Staphylococcus aureus, and Rhizoctonia solani stored in a -80℃ refrigerator on the corresponding solid culture medium. Then seal the culture dish with a pre-sterilized sealing film. Put it in a constant temperature incubator at a suitable temperature for growth.

[0037] Cultivation: Take each newly grown bacterial strain and place it in a fresh sterilized liquid culture medium, and put glass beads of appropriate size in the bottle (to place bacterial clumps), seal the bottle mouth, and place it in a shaker at a suitable temperature for cultivation. The steps for culturing bacterial strains in solid culture medium are the same as those for activation.

[0038] 3. Antibacterial reaction operation and results

[0039] The antibacterial effects of the five products in Table 1 were studied using the antibacterial paper disc method test. Escherichia coli, Staphylococcus aureus, yeast X33, and yeast JYM789 were used as research objects, and the antibacterial effect of ampicillin was used as a positive control. The above products were dissolved in DMSO and prepared into solutions of the same concentration. Prepare filter paper pieces of the same size, 10ul pipette tips, and glass beads and sterilize them at 115℃ for 25 minutes. Use a pipette to draw up 20ul of the liquid culture medium solution with bacteria, drop it on the solid culture medium, place glass beads and roll them to distribute the colonies as evenly as possible, then place an appropriate amount of filter paper pieces on the culture medium to fix and number them, and use a pipette to draw up 5 microliters of the product solution corresponding to the number and drop it on the filter paper. After the addition is complete, seal the culture dish and put it in a constant temperature incubator for growth. The results are as follows Figure 1 and Figure 2As shown, when the bacterial species is Escherichia coli or Staphylococcus aureus, only the antibacterial effect of ampicillin is observed, and the above products have no antibacterial effect; Figure 3 and Figure 4 As shown, when the bacterial species are yeast X33 and yeast YJM789, ampicillin and the above products have no antibacterial effect.

[0040] The mycelium growth rate method was used to study the inhibitory effects of the five products in Table 1 on the growth of the plant pathogen Rhizoctonia solani. Fresh bacterial discs were prepared from the edge of the Rhizoctonia solani colonies cultured on PDA medium for 4 to 5 days and inoculated into culture media containing a series of product concentrations. The concentrations were designed to be 4 mg / mL, 2 mg / mL, 1 mg / mL, 0.5 mg / mL, and 0.005 mg / mL. They were cultured in a 27°C incubator, and the colony diameters were measured after the blanks were fully grown. It was found that three products had a strong inhibitory effect on the growth of the plant pathogen Rhizoctonia solani, which are Product A, Product B, and Product C in Table 1. Figure 5 and Figure 6 As shown in Figure 2, when the concentration of product A and product B was 2 mg / mL, no mycelial growth was observed; Figure 7 As shown in the figure, when the concentration of product C is 1 mg / mL, the growth diameter of mycelium is only 0.2 cm, while products D and E also have a certain inhibitory effect on the growth of plant pathogenic bacteria-Rhizoctonia solani during the antibacterial effect experiment, but the antibacterial effect is not ideal. In addition, the antibacterial effect comparison of conventional antibacterial compounds against plant pathogens and product C is as follows: Figure 8 As shown, it is indicated that the three compounds with specific structures (product A, product B and product C) selectively have a strong inhibitory effect on the growth of the plant pathogen Rhizoctonia solani, and can be further used to prepare antibacterial drugs against the plant pathogen Rhizoctonia solani.

[0041] The above embodiments describe the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The above embodiments and descriptions are only for illustrating the principles of the present invention. Without departing from the scope of the principles of the present invention, the present invention may have various changes and improvements, and these changes and improvements all fall within the scope of protection of the present invention.

Claims

1. Application of fully substituted 1,2,3-triazole-4-phosphonate compounds in the preparation of antibacterial drugs against plant pathogens, characterized in that: The structural formula of the fully substituted 1,2,3-triazole-4-phosphine ester compound is: The plant pathogen is Rhizoctonia solani.

2. The use according to claim 1, characterized in that: The concentration of the fully substituted 1,2,3-triazole-4-phosphonate compound in the bacterial solution is 0.5-4 mg / mL.

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