Benzimidazolyl-containing isopropanolamine compound as well as preparation method and application thereof

Abstract

The invention relates to benzimidazolyl-containing isopropanolamine compounds as well as a preparation method and application thereof. The compound has a structure as shown in a general formula (I): # imgabs0 #, 2-mercapto benzimidazole is taken as a basis, a nitrogen-containing fragment is introduced into the system, a series of substituted benzimidazolyl isopropanolamine compounds are synthesized, and the compounds have an excellent inhibition effect on plant pathogenic bacteria.

Description

Technical Field

[0001] The invention relates to the technical field of pharmaceutical chemistry, in particular to an isopropanolamine compound containing benzimidazole and a preparation method and application thereof. Background Art

[0002] In recent years, plant bacteria and fungi have seriously affected the yield and quality of crops around the world, causing huge economic losses to farmers. For example, Xanthomonas oryzae pV.Oryzae is a rod-shaped Gram-negative bacterium that can cause rice leaves to wilt and turn white, causing at least 10-50% reduction in yield in rice-growing countries every year. In addition, Xanthomonas axonopodis pV.Citri can cause citrus rot, affecting citrus yields worldwide. In the process of agricultural production, due to the long-term use of traditional agents, plant pathogens have developed a certain resistance to them. Therefore, it is of great significance to create new, efficient, low-toxic and safe green pesticides.

[0003] Benzimidazole is a common drug intermediate, which is widely used in the fields of medicine and pesticides and has broad development potential. According to literature reports, benzimidazole has a wide range of biological activities, such as antibacterial, antiviral, anti-inflammatory, anticancer, etc. In order to find highly effective antibacterial active compounds, the present invention is based on the structure of 2-mercaptobenzimidazole, uses isopropanol as a connecting skeleton, introduces a series of nitrogen-containing saturated aliphatic heterocycles or aliphatic secondary amine groups into this system, synthesizes a series of isopropanolamine compounds containing benzimidazole, examines their biological activities, and provides an important scientific basis for the research and development and creation of new pesticides.

[0004] The research progress of the biological activity of benzimidazole compounds is as follows:

[0005] In 2024, Liu et al. (Liu, J.; Chen, H.; LV, Y.; Wu, H.; Yang, L.-j.; Zhang, J.; Huang, J.; Wang, W., Synthesis, Characterization, and Antifungal Activity of Benzimidazole-Grafted Chitosan against Aspergillus flaVus. Journal of Agricultural and Food Chemistry 2024, 72(19), 11185-11194.) synthesized a new benzimidazole-grafted chitosan (BAC) by connecting chitosan (CS) with a simple benzimidazole compound 2-benzimidazole propionic acid (BA). The characterization of BAC was performed by Fourier transform infrared (FTIR) spectroscopy and nuclear magnetic resonance spectroscopy ( 1 H and 13 C NMR). Then, the efficiency of BAC against A. flavus ACCC 32656 was investigated in terms of spore germination, mycelial growth, and aflatoxin production. BAC exhibited better antifungal effects than CS and BA. The minimum inhibitory concentration (MIC) value of BAC was 1.25 mg / mL, while neither CS (16.0 mg / mL) or BA (4.0 mg / mL), which had the highest solubility, could completely inhibit the growth of A. flavus. In addition, the results showed that BAC inhibited spore germination and elongation by affecting ergosterol biosynthesis and cell membrane integrity, leading to plasma membrane permeabilization and leakage of intracellular contents. Aflatoxin production was also inhibited when treated with BAC. These findings suggest that benzimidazole-derived natural CS has the potential to be used as an ideal antifungal agent for food preservation.

[0006] In 2024, Lu et al. (Lu, S.; Liang, B.; Hu, J.; Liu, Y; Yang, F.; Liu, J., Multidimensional response of dopamine nano-system for on-demand fungicides delivery: Reduced toxicity and synergistic antibacterial effects. Chemical Engineering Journal 2024, 482.) used mesoporous polydopamine nanoparticles (MPDA NPs) as carriers, prochloraz (Pro) as loading agents, and benzimidazole (BIZ) and β-cyclodextrin (β-CD) complexes as blocking agents to prepare on-demand delivery nanopesticide Pro@MPDA-BIZ@β-CD (hereinafter referred to as PMBD), which can be applied in stages. Sclerotinia sclerotiorum secretes oxalic acid factor to create an acidic microenvironment, and the nanosystem quickly releases the fungicide Pro into the mycelium. The MPDA material also reacts with oxalic acid to jointly reduce the pathogenicity of Sclerotinia sclerotiorum, effectively inhibit the growth of Sclerotinia sclerotiorum, and achieve the treatment of Sclerotinia sclerotiorum infection. On the other hand, when the plants were not infected with Sclerotinia sclerotiorum, MPDA with photothermal effect triggered the release of fungicides under near-infrared light irradiation, thereby preventing Sclerotinia sclerotiorum infection. In vitro release experiments showed that the cumulative release of Pro under low pH and near-infrared light irradiation was 2.68 times and 2.04 times that under neutral and lightless conditions, respectively. On the other hand, the rich functional groups on MPDA-BIZ@β-CD (hereinafter referred to as MBD) endowed the system with good wettability. The contact angles of MBD on Chinese cabbage and tomato leaves were reduced by 14.85° and 41.54°, respectively, enhancing their ability to resist rain erosion and preventing the loss of fungicides into the environment. In addition, biosafety experiments showed that PMBD had good biocompatibility with plant germination and soil microorganisms.

[0007] In 2024, Maryam et al. (Kia, M.; Ghassemzadeh, M.; Eslami Moghadam, M.; Saeidifar, M.; Mohsenzadeh, F.; Fuhr, O.; Fenske, D., Novel copper-drugs bearing dipodal bis-mercaptobenzimidazoles: Synthesis, crystal structures, in Vitro biological activities, DNA binding, DFT calculations and molecular docking. Journal of Molecular Liquids 2024, 413.) successfully prepared three new copper (II) complexes 1-3 with dipodal bis-mercaptobenzimidazole derivatives based on o-xylene, m-xylene or p-xylene (L1, L2 and L3) and characterized them using various spectroscopic techniques, including elemental analysis, FT-IR, 1H NMR and UV-Vis spectra, LC-MS spectra, and TGA. The analysis showed that the ligand / metal molar ratio in all copper(II) complexes was 1:1, and the ligands coordinated to the metal center as neutral N donors. In addition, the crystal structures of L1, L2, L3, and copper(II) complex 2 were determined using single crystal X-ray diffraction (SC-XRD) analysis. Copper(II) complexes 1-3 showed remarkable stability in the pH range of 4-10. The antibacterial effect was tested against Gram-negative (Escherichia coli, E. coli ATCC 25922 PTCC 1399) and Gram-positive (Staphylococcus aureus, Staphylococcus aureus ATCC6538 PTCC 1112) bacterial strains. It should be noted that the complexes showed enhanced antibacterial activity (up to 99%) compared to the free ligand. In vitro studies of all synthesized compounds included testing against human colorectal cancer cell line (HCT-116) using MTT assay. The results showed that the copper (II) complexes exhibited lower CC50 values ​​(CC50 ranged from 0.045 mM to 0.135 mM) compared to their corresponding ligands (CC50 ranged from 0.150 mM to 0.240 mM) and carboplatin (CC50 = 0.165 mM), indicating that the anticancer activity of the complexes was higher. In addition, DAPI staining and fluorescence spectroscopy (at three different temperatures) were performed to further examine the effects of complexes 1-3 on the induction of cell apoptosis and to explore their interactions with DNA, respectively. The results revealed a dynamic quenching mechanism, with hydrophobic forces playing a dominant role in the binding process. Viscosity measurements showed that all complexes could interact with DNA in a groove-binding manner. Density functional theory (DFT) calculations were used to support structural and vibrational studies and predict chemical reactivity. Docking simulations were performed to evaluate the behavior of their synthesized compounds towards DNA (PDB: 1BNA). These results suggest that various elements, including NH groups, imidazole rings, thioether sulfur atoms, and sulfate moieties of ligands and complexes, are involved in the groove binding with DNA.

[0008] In 2024, Yang et al. (Yang, S.; Zhang, J.-R.; Li, M.; Feng, Y.-M.; Yang, L.-l.; Long, Z.-Q.; Zhou, X.; Wu, Z.-B.; Liu, L.-W.; Yang, S., Expanding the Structural DIVersity of Tubulin-Targeting Agents: DeVelopment of Highly Potent Benzimidazoles for Treating Fungal Diseases. Journal of Agricultural and Food Chemistry 2024, 72(28), 15541-15551.) prepared new benzimidazole derivatives by introducing attractive pyrimidine pharmacophores. 2-((6-(4-(trifluoromethyl)phenoxy)pyrimidin-4-yl)thio)-1H-benzo[d]imidazole (A25) showed the best antifungal activity against Sclerotinia sclerotiorum (Ss), with a maximum inhibitory effective concentration (EC 50 ) was 0.158 μg / mL, which was higher than the reference agent carbendazim (EC 50 =0.594 μg / mL). Pot experiments showed that compound A25 (200 μg / mL) had acceptable protective activity (84.7%) and therapeutic activity (78.1%), which was comparable to that of carbendazim (protective activity: 90.8%; therapeutic activity: 69.9%). Molecular docking showed that A25 could form multiple hydrogen bonds and π-π interactions with β-tubulin, and its bonding effect was stronger than that of carbendazim. Fluorescence imaging showed that the intracellular microtubule structure changed significantly after A25 treatment. Overall, these remarkable antifungal properties of the constructed novel benzimidazole derivatives can promote the application of novel microtubule targeting agents.

[0009] In 2022, Yang et al. (Yang, X.; Syed, R.; Fang, B.; Zhou, CH, A New DiscoVerytowards NoVel Skeleton of Benzimidazole-Conjugated Pyrimidinones as UniqueEffectIVe Antibacterial Agents. Chinese Journal of Chemistry 2022, 40(22), 2642-2654.) used urea, ethyl 4-chloroacetoacetate and various aldehydes as raw materials, and developed a new class of potential antibacterial agents with a unique pyrimidone benzimidazole skeleton through nucleophilic substitution and Biginelli reaction. Some target molecules showed strong antibacterial activity, especially the pyrimidone benzimidazole hybrid 9e, which had the strongest growth inhibition effect on Enterococcus faecalis and Pseudomonas aeruginosa, and the MIC value was lower than 1 μg / mL of norfloxacin. In addition, compound 9e showed strong anti-biofilm ability, low drug resistance and excellent biosafety against human erythrocytes. Further studies showed that compound 9e could disrupt membrane integrity and lead to leakage of cellular components such as proteins and nucleic acids. At the same time, compound 9e could reduce lactate dehydrogenase activity, block cellular metabolism, and intercalate with DNA. ADMET analysis predicted that molecule 9e had good pharmacokinetic properties and good bioavailability characteristics. Summary of the invention

[0010] One of the purposes of the present invention is to provide an isopropanolamine compound containing benzimidazole or its stereoisomer, or its salt or solvate.

[0011] Another object of the present invention is to provide an intermediate compound for preparing the above-mentioned compound or its stereoisomer, or its salt or solvate and a preparation method thereof.

[0012] Another object of the present invention is to provide a composition containing the above compound or its stereoisomer, or its salt or solvate.

[0013] Another object of the present invention is to provide the use of the above compound or its stereoisomer, or its salt or solvate, or the composition.

[0014] Another object of the present invention is to provide a method for controlling agricultural pests and diseases using the above-mentioned compound or its stereoisomer, or its salt or solvate, or the composition.

[0015] To achieve the above object, the present invention adopts the following technical solutions:

[0016] A benzimidazole isopropanolamine compound or a stereoisomer thereof, or a salt thereof or a solvate thereof, wherein the compound has a structure as shown in the general formula (I):

[0017]

[0018] Wherein, R1 and R2 are selected from one or more of an optionally substituted or unsubstituted alkyl group and an aryl group, and an optionally substituted or unsubstituted heteroaryl group;

[0019] Preferably, R1 and R2 are selected from methyl, benzyl, 2-methoxybenzyl, 3-methoxybenzyl, 4-methoxybenzyl, 3,4-dimethoxybenzyl, 2-fluorobenzyl, 3-fluorobenzyl, 4-fluorobenzyl, 4-trifluoromethylbenzyl, 2-methylbenzyl, 3-chlorobenzyl, 4-chlorobenzyl, 4-(4-pyridyl)piperazine. Most preferably, R1 is selected from hydrogen and R2 is selected from 4-chlorobenzyl.

[0020] The present invention also provides an intermediate compound for preparing the benzimidazole-containing isopropanolamine compound or its stereoisomer, or its salt or solvate:

[0021]

[0022] The present invention also provides a method for preparing the isopropanolamine compound containing benzimidazole or its stereoisomer, or its salt or solvate, comprising the following steps:

[0023]

[0024]

[0025] wherein R1 and R2 are as described above.

[0026] The present invention also provides a composition, which contains the compound or its stereoisomer, or its salt or solvate, and an agriculturally usable adjuvant or fungicide, insecticide or herbicide; preferably, the composition is in the form of emulsifiable concentrate (EC), dust (DP), wettable powder (WP), granule (GR), aqueous solution (AS), suspension (SC), ultra low volume spray (ULV), soluble powder (SP), microcapsule (MC), smoke agent (FU), emulsion in water (EW), water dispersible granule (WG).

[0027] The compound or its stereoisomer, or its salt or solvate, or the composition can be used to prevent and control agricultural pests and diseases. Preferably, the agricultural pests and diseases are bacterial or fungal plant diseases; more preferably, the agricultural pests and diseases are plant leaf blight and plant canker; most preferably, the agricultural pests and diseases are rice bacterial leaf blight, cucumber bacterial leaf blight, konjac bacterial leaf blight, citrus canker, grape canker, tomato canker, kiwi canker, apple canker, cucumber gray mold, pepper wilt pathogen, rapeseed sclerotinia, wheat fusarium head blight, potato late blight, and blueberry root rot.

[0028] The present invention also provides a method for preventing and controlling agricultural pests and diseases, wherein the compound or its stereoisomer, or its salt or solvate, or the composition acts on harmful substances or their living environment; preferably, the agricultural pests and diseases are bacterial or fungal plant diseases; more preferably, the agricultural pests and diseases are rice bacterial blight, tobacco bacterial wilt, cucumber bacterial blight, konjac bacterial blight, citrus canker, grape canker, tomato canker, kiwi canker, apple canker, cucumber gray mold, pepper wilt pathogen, rapeseed sclerotinia, wheat fusarium head blight, potato late blight, and blueberry root rot.

[0029] The present invention also provides a method for protecting plants from agricultural pests and diseases, which comprises a method step in which the plant is contacted with the compound or its stereoisomer, or its salt or solvate, or the composition.

[0030] The term "substituted" as used herein refers to the replacement of any one or more hydrogen atoms on a designated atom or group with a selected designated group, provided that the general valence of the designated atom is not exceeded. If not otherwise specified, the substituents are named to the central structure. For example, it is understood that when (cycloalkyl)alkyl is a possible substituent, the point of attachment of the substituent to the central structure is in the alkyl portion. A ring double bond as used herein is a double bond formed between two adjacent ring atoms (such as C=C, C=N or N=N). When referring to substitution, especially polysubstitution, it means that multiple substituents are substituted at various positions on the designated group, such as dichlorophenyl refers to 1,2-dichlorophenyl, 1,3-dichlorophenyl and 1,4-dichlorophenyl.

[0031] When a substituent is referred to as an alkylaryl group or a heteroarylamino group, or when these substituents are specifically a certain alkylaryl group or heteroarylamino group, it refers to one to three of the above substituents. For example, methylphenyl refers to a phenyl group substituted with one to three methyl groups.

[0032] By adopting the above technical scheme, the present invention synthesizes a series of benzimidazole compounds containing isopropanolamine substructures based on benzimidazole compounds, and finds that the compounds have good inhibitory effects on pathogenic bacteria, such as rice bacterial blight pathogen (Xanthomonas oryzae pV.oryzae, Xoo), citrus canker pathogen (Xanthomonas axonopodis pV.citri, Xac), etc., and have good inhibitory effects on pathogenic fungi, providing an important scientific basis for the research and development and creation of new pesticides. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 It is a comparison chart of the protective activity and therapeutic activity of compound 18 of the present application and related compounds. Example

[0034] The present invention is further described below by way of examples. It should be understood that the methods described in the examples of the present invention are merely for illustrating the present invention, rather than limiting the present invention, and simple improvements to the preparation methods of the present invention under the premise of the concept of the present invention all fall within the scope of the present invention. All raw materials and solvents used in the examples are commercially available products.

[0035] Example 1: Preparation of Intermediate (R)-3,4-dihydro-2H-benzo[4,5]imidazole[2,1-b][1,3]thiazin-3-ol (II) or (S)-3,4-dihydro-2H-benzo[4,5]imidazole[2,1-b][1,3]thiazin-3-ol (IV)

[0036] 2-Mercaptobenzimidazole (1000 mg, 6.52 mmol) and NaOH (417.6 mg, 10.44 mmol) were added to a round-bottom flask containing 30 mL of anhydrous ethanol, stirred and dissolved, and then (R)-(-)-epichlorohydrin or (S)-(+)-epichlorohydrin (510.29 μL, 6.52 mmol) was added, and stirred and refluxed at 79°C for 5.0 h, and the reaction progress was monitored by a thin layer chromatography plate. After the reaction was completed, the mixed system was washed with ice water and filtered under reduced pressure to obtain a purified intermediate II or IV (properties: white powder, yield: 94.37% (II) or 93.70% (IV)). Its NMR data are: Intermediate (II): 1 H NMR (400MHz, DMSO-d6) δ7.50-7.38 (m, 2H, Phenyl-H), 7.18-7.10 (m, 2H, Phenyl-H), 5.78 (s, 1H, -OH), 4.47 (dq, J=6.1, 3.1Hz, 1H, CH-OH), 4.23 (dd, J=12.7, 3.6Hz, 1H, N- CH2 ), 4.08 (dd, J=12.6, 5.6Hz, 1H, N- CH2 ), 3.40 (dd, J=12.7, 2.5Hz, 7H, DMSO-H2O and S- CH2 ), 3.21 (dd, J=12.5, 6.9Hz, 1H, S- CH2 ). 13 C NMR (101 MHz, DMSO-d6) δ 146.27, 142.82, 135.95, 121.79, 120.84, 117.00, 108.74, 60.63, 48.24, 31.40. Intermediate (IV): 1 H NMR (400MHz, DMSO-d6) δ7.48-7.43 (m, 1H, Phenyl-H), 7.43-7.38 (m, 1H, Phenyl-H), 7.17-7.12 (m, 2H, Phenyl-H), 5.80 (s, 1H, -OH), 4.47 (tt, J=6.1, 2.9Hz, 1H, CH -OH), 4.23 (dd, J=12.7, 3.6Hz, 1H, N- CH2 ), 4.08 (dd, J=12.7, 5.6Hz, 1H,, N- CH2 ), 3.41 (d, J=2.5Hz, 1H, S- CH2 ), 3.21 (dd, J=12.6, 6.9Hz, 1H S- CH2 ). 13 C NMR (101MHz, DMSO-d6) δ146.27, 142.82, 135.95, 121.79, 120.84, 117.01, 108.75, 60.63, 48.24, 31.41.

[0037] Example 2: Preparation of intermediate (3R)-3-(oxy-2-methoxy)-3,4-dihydro-2H-benzo[4,5]imidazole[2,1-b][1,3](III) or (3S)-3-(oxy-2-methoxy)-3,4-dihydro-2H-benzo[4,5]imidazole[2,1-b][1,3](V)thiazine

[0038] Intermediate II or IV (1000 mg, 4.84 mmol) and KOH (544.02 mg, 9.70 mmol) were dissolved in DMF (15 mL), stirred for 15 min, and then epibromopropane (625 μL, 7.76 mmol) was added. The reaction was allowed to react at room temperature for 5.0 h before the reaction was terminated. The reaction system was extracted with saturated NH4Cl and ethyl acetate, and the organic phase was retained and distilled under reduced pressure to obtain a preliminarily purified intermediate 3. Column chromatography (eluent: V 二氯甲烷 :V 甲醇 =30:1.0) to further obtain the final purified intermediate III or V (property: white powder, yield: 78.30% (III) or 88.52% (V)). Its NMR data are: Intermediate III: 1 H NMR (500MHz, Chloroform-d) δ7.61 (dt, J=6.7, 1.1Hz, 1H, Phenyl-H), 7.25-7.17 (m, 3H, Phenyl-H), 4.41-4.32 (m, 1H, CH2- CH -O(CH2)), 4.32-4.25(m, 1H, OCH- CH2 -N), 4.19-4.11 (m, 1H, OCH- CH2 -N), 4.05 (ddd, J=38.6, 11.8, 2.3Hz, 1H, O- CH2 ), 3.53 (ddd, J=61.4, 11.9, 6.2Hz, 1H, O- CH2 ), 3.39-3.27 (m, 2H, OCH- CH2 -S), 3.20 (dtd, J=10.8, 6.7, 2.5Hz, 1H, OCH2- CH -O), 2.86-2.83 (m, 1H, OCH- CH2 -O), 2.67-2.64 (m, 1H, OCH- CH2 -O). 13 C NMR (126 MHz, Chloroform-d) δ 143.55, 135.93, 122.65, 121.74, 118.40, 118.37, 107.95, 107.90, 103.05, 70.73, 70.43, 70.19, 50.96, 50.93, 47.09, 46.57, 44.31, 44.23, 29.43, 28.85. Intermediate V: 1H NMR (500MHz, Chloroform-d) δ7.48 (d, J=7.9Hz, 1H, Phenyl-H), 7.10 (t, J=7.5Hz, 1H, Phenyl-H), 7.06-6.96 (m, 2H, Phenyl-H), 4.16-4.06 (m, 1H, CH2- CH -O(CH2)), 3.96(d, J=4.8Hz, 1H, OCH- CH2 -N), 3.92(t, J=4.3Hz, 1H, OCH- CH2 -N), 3.82 (ddd, J=38.6, 11.8, 2.4Hz, 1H, O- CH2 ), 3.30 (ddd, J=57.3, 11.8, 6.2Hz, 1H, O- CH2 ), 3.19-3.05 (m, 2H, OCH- CH2 -S), 3.00 (tdt, J=6.6, 4.6, 2.5Hz, 1H, OCH2- CH -O), 2.66(q, J=4.2Hz, 1H, OCH- CH2 -O), 2.47(dt, J=4.7, 2.2Hz, 1H, OCH- CH2 -O). 13 CNMR (126MHz, Chloroform-d) δ145.87, 145.81, 142.86, 142.83, 135.49, 135.47, 122.10, 121.22, 117.52, 11 7.50, 107.75, 107.71, 69.80, 69.67, 69.54, 69.05, 50.48, 50.46, 46.47, 46.05, 43.85, 43.76, 28.74, 28.22.

[0039] Example 3: Preparation of 1-((4-chlorobenzyl)amino)-3-(((R)-3,4-dihydro-2H-benzo[4,5]imidazo[2,1-b][1,3]3-thiazinyl)oxy)propan-2-ol

[0040] Intermediate III (300 mg, 1.14 mmol), K2CO3 (110.64 mg, 0.8 mmol) and isopropanol solution (10 mL) were added to a 15 mL pressure tube, stirred at 60 ° C for 10 min, and then p-chlorobenzylamine (1.72 mmol) was added and the reaction was continued for 4.0 h. After the reaction was completed, the reaction system was extracted with water and dichloromethane, and the organic layer was retained and dried by vacuum distillation to obtain preliminarily purified BiTAl8. Column chromatography purification (eluent: V 二氯甲烷 :V甲醇 =10:1.0), a light yellow oily liquid was obtained, with a yield of 83.18%. 1 H NMR, 13 The structures were characterized by C NMR and HRMS.

[0041] Example 4: Preparation of binary compound 18@β-CD

[0042] The tetrahydrofuran solution containing the guest molecule 18 (8.0 μL, 0.124 M) was gradually dripped into 2.0 mL of deionized water containing the host molecule β-CD (0.496 mM). After THF was naturally evaporated, the self-assembled supramolecular binary complex 18@β-CD was formed.

[0043] Other compounds were prepared by similar methods to those of Examples 1-3, replacing corresponding starting materials.

[0044] The structures, hydrogen NMR spectra, carbon NMR spectra, high-resolution mass spectrometry data and physicochemical properties of the synthesized substituted phenyl isopropanolamine compounds are shown in Table 1.

[0045] Table 1 H NMR and C NMR data of compounds

[0046]

[0047]

[0048]

[0049]

[0050]

[0051]

[0052]

[0053]

[0054] Pharmacological Example 1:

[0055] EC 50 (median effect IVe concentration) is an important indicator for evaluating the sensitivity of plant pathogens to compounds. It is also an important parameter for setting compound concentration when studying the mechanism of action of target compounds. In the concentration gradient experiment, the two-fold dilution method was used to set 6 appropriate concentrations. Finally, the inhibition rate of the agent on plant pathogens and the concentration of the agent were converted into logarithmic values. The toxicity curve was obtained through regression analysis with SPSS software, and the EC was calculated. 50 .

[0056] The effective medium concentration EC of the target compound against plant pathogens was tested by turbidity method. 50 , the test subjects were Xanthophyton solani (Xoo) and Xanthophyton citri citri (Xac). Use the two-fold dilution method to set 6 appropriate concentrations (for example: 50, 25, 12.5, 6.25, 3.125, 1.5625 μg / mL), DMSO as the blank control, thiophanate-methyl (TC) and kasugamycin (KSM) as positive controls. Then add the drugs to the NB culture medium according to the concentration gradient, and then add 40 μL of bacterial solution in the logarithmic growth phase to the drug-containing culture medium, and culture in a constant temperature shaker at 28°C and 220 rpm. Until the turbidity of the blank control (OD 595nm ) reached 0.6, and then the culture was stopped. Finally, the OD of each concentration of bacterial solution was measured on an ELISA reader. 595nm The OD of the blank control was also measured. 595nm The inhibition rate of the pesticide on plant pathogens and the concentration of the pesticide were converted into logarithmic values, and the toxicity curve was obtained through regression analysis with SPSS software to calculate the corresponding EC 50 value.

[0057] The inhibition rate calculation formula is as follows:

[0058]

[0059] in

[0060] Corrected OD value = OD value of culture medium - OD value of sterile culture medium

[0061] The embodiments of the present invention are used to illustrate the technical solutions of the present invention, but the contents of the embodiments are not limited thereto. The experimental results of the target compounds are shown in Tables 2 to 5.

[0062] Table 2 Preliminary screening of the activity of the compounds of the present application against plant pathogenic bacteria Xoo

[0063]

[0064] Table 3 Preliminary screening of the activity of the compounds of the present application against plant pathogenic bacteria Xac

[0065]

[0066]

[0067] Table 4 EC values ​​of some compounds of this application against plant pathogenic bacteria Xoo 50 value

[0068]

[0069] Table 5 EC values ​​of some compounds of this application against plant pathogenic bacteria Xac 50 value

[0070]

[0071] a: EC50 values ​​of antibacterial activity are expressed as mean + SD (standard deviation). b: Commercial fungicides are used as positive controls, TC (thiophanate-methyl), KSM (kasugamycin).

[0072] As can be seen from Tables 2 to 5, in the in vitro test, the target compounds showed good inhibitory activity against plant pathogens (such as Xanthomonas oryzae pV.oryzae, Xoo). The compounds showed good inhibitory activity against Xanthomonas oryzae pV.oryzae, among which the EC values ​​of compounds 5, 10, 11, 17, 18, 22, 23, 24, 25, and 26 were 50 The EC values ​​of compounds 5, 10, 11, 17, 18, 22, 24, 25, and 26 were 12.8, 18.8, 2.84, 32.1, 2.25, 45.6, 27.8, 23.3, 25.9, and 18.2 μg / mL, respectively. It also showed excellent inhibitory activity against citrus canker pathogen (Xanthomonas axonopodis pV.citri, Xac). 50 They are 6.43, 27.1, 4.01, 16.8, 4.74, 19.0, 9.95, 18.1 and 18.2 respectively, and can be used to prepare pesticides against plant pathogenic bacteria.

[0073] Pharmacological Example 2:

[0074] Compound 18 showed good in vitro activity against Xanthomonas oryzae (EC 50 2.25 μg / mL), and then the bioactivity of the drug against Xanthomonas oryzae in vivo was tested by leaf cutting method. The specific method is as follows: Use sterilized scissors to dip into the leaf containing Xanthomonas oryzae (OD 595=0.6-0.8) of bacterial solution was inoculated on rice plants (rice variety: Fengyouxiangzhan, cultivation time: 8 weeks), and three replicates were used for each treatment. For protective activity, 200 μg / mL of drug solution (compound 18, 18@β-CD, 18@γ-CD, β-CD, γ-CD and positive control drug thiophanate-methyl (20% content preparation), kasugamycin (98% active ingredient)) was evenly sprayed on rice leaves until droplets fell, and an equal amount of DMSO aqueous solution and bacterial leaves without drug were set as controls, and the bacteria were inoculated after 24 hours; for therapeutic activity, the bacteria were first inoculated on rice plants, and 200 μg / mL of drug solution was evenly sprayed on rice leaves after 24 hours. Finally, the treated samples were placed in an artificial climate chamber (temperature: 28°C, humidity: 90%) for 14 days, and the length and total length of the lesions on the rice leaves were recorded, and the corresponding prevention and control effects were obtained by the grading standard calculation method.

[0075] The grading standard is as follows: First, the diseased length of each leaf and the total length of the entire leaf are measured, and then the percentage of the diseased length and the total length is calculated. Secondly, the leaves are classified according to the following grade standards: Grade 1, the diseased length is less than 5% of the total leaf length; Grade 3, the diseased length accounts for 6-10% of the total length; Grade 5, the diseased length accounts for 11-20% of the total length; Grade 7, the diseased length accounts for 21-50% of the total length; Grade 9, the diseased length accounts for more than 50% of the total length. Among them: β-CD represents β-cyclodextrin, γ-CD represents γ-cyclodextrin.

[0076] The calculation method of the disease index is as follows:

[0077] Disease index = ∑ (number of leaves in each level × corresponding level) / (total number of leaves × highest level)

[0078] The calculation method of protection effect is as follows:

[0079] Preventive effect (%) = (disease index of control group - disease index of treatment group) / disease index of control group × 100

[0080] The examples of the present invention are used to illustrate the technical solution of the present invention, but the implementation content is not limited thereto. The experimental results of the target compounds are shown in Table 6, Figure 1 shown.

[0081] Table 6 Control effects of compounds 18, 18@β-CD, 18@γ-CD, β-CD, and γ-CD on rice bacterial blight

[0082]

[0083] As shown in Table 6, β-CD and γ-CD have almost no control effect on rice bacterial blight, and compound 18 has considerable protective activity (42.68%) and therapeutic activity (35.90%), which is lower than its binary compound 18@β-CD (protective and therapeutic activities are 54.39% and 49.86%, respectively) and 18@γ-CD (protective and therapeutic activities are 46.70% and 43.94%, respectively). Their therapeutic and protective activities are slightly better than the positive control drugs thiophanate-methyl and kasugamycin.

Claims

1. A class of substituted benzimidazole-containing isopropanolamine compounds or their stereoisomers, or their salts or solvates, characterized in that The compound has a structure as shown in general formula (I): Wherein, R1 and R2 are independently selected from one or more of any substituted or unsubstituted alkyl, aryl, alicyclic, and heteroaryl groups.

2. The substituted benzimidazole-containing isopropanolamine compound or its stereoisomer, or its salt or solvate according to claim 1, characterized in that: R1 and R2 are independently selected from hydrogen, methyl, ethyl, propyl, isopropyl, benzyl, 2-methoxybenzyl, 3-methoxybenzyl, 4-methoxybenzyl, 2-fluorobenzyl, 3-fluorobenzyl, 4-fluorobenzyl, 2-chlorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl, 2-methylbenzyl, 3-methylbenzyl, 4-methylbenzyl, 2-trifluoromethylbenzyl, 3-trifluoromethylbenzyl, 4-trifluoromethylbenzyl, 2,3-dimethoxybenzyl, 3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl, 2,5-dimethoxybenzyl, 2,6-dimethoxybenzyl, tetrahydropyrrole, piperazine, 4-(4-pyridyl)piperazine.

3. The substituted benzimidazole-containing isopropanolamine compound according to claim 1, or its stereoisomer, or its salt or solvate, characterized in that Selected from the following specific compounds:

4. An intermediate compound for preparing the benzimidazole-containing isopropanolamine compound or its stereoisomer, or its salt or solvate according to claim 1, characterized in that As shown below:

5. A method for preparing a benzimidazole-containing isopropanolamine compound or a stereoisomer thereof, or a salt thereof or a solvate thereof according to any one of claims 1 to 3, characterized in that The steps include: Wherein R1 and R2 are as described in any one of claims 1-3.

6. A composition characterized in that Containing the compound or its stereoisomer according to any one of claims 1 to 4, or its salt or solvate, and an agriculturally usable adjuvant or fungicide, insecticide or herbicide; preferably, the composition is in the form of emulsifiable concentrate (EC), dust (DP), wettable powder (WP), granule (GR), aqueous solution (AS), suspension (SC), ultra low volume spray (ULV), soluble powder (SP), microcapsule (MC), smoke agent (FU), emulsion in water (EW), water dispersible granule (WG); preferably, the adjuvant is β-cyclodextrin or γ-cyclodextrin.

7. Use of the compound or its stereoisomer, or its salt or solvate according to any one of claims 1 to 4, or the composition according to claim 6 for preventing and controlling agricultural pests and diseases, preferably, the agricultural pests and diseases are bacterial or fungal diseases of plants; more preferably, the agricultural pests and diseases are plant leaf blight and plant canker; most preferably, the agricultural pests and diseases are rice bacterial leaf blight, cucumber bacterial leaf blight, konjac bacterial leaf blight, citrus canker, grape canker, tomato canker, kiwi canker, apple canker, cucumber gray mold, pepper wilt pathogen, rapeseed sclerotinia, wheat fusarium head blight, potato late blight, blueberry root rot.

8. A method for preventing and controlling agricultural pests and diseases, characterized in that: The compound or stereoisomer thereof, or salt or solvate thereof according to any one of claims 1 to 4, or the composition according to claim 6 is allowed to act on a pest or its living environment; preferably, the pest is a plant bacterial or fungal disease; more preferably, the agricultural pests and diseases are rice bacterial blight, tobacco bacterial wilt, cucumber bacterial blight, konjac bacterial blight, citrus canker, grape canker, tomato canker, kiwi canker, apple canker, cucumber gray mold, pepper wilt pathogen, rapeseed sclerotinia, wheat fusarium head blight, potato late blight, and blueberry root rot.

9. A method for protecting plants from agricultural pests and diseases, comprising a method step in which the plant is contacted with a compound or stereoisomer thereof, or a salt or solvate thereof, or a composition according to claim 6.

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