Use of a benzene-1,2,4-triazineoxazepine compound

Abstract

The application discloses application of a benzene-1,2,4-triazine and oxazepine compound, and belongs to the technical field of insecticide preparation. The benzene-1,2,4-triazine and oxazepine compound is used in the preparation of insecticides, the compound has high enzyme inhibition activity on insect chitinase, can exhibit excellent insecticidal effect, and has high inhibition activity on Asian corn borer chitinase (OfChi-h) in particular; the compound has strong insecticidal activity on agricultural pests such as Asian corn borer and diamondback moth. The compound with the structure is used in the preparation of insecticides, has high pesticide research value, and has wide application prospects in the field of pesticide science.

Description

Technical Field

[0001] This invention belongs to the field of insecticide preparation technology, and more specifically relates to a benzo-1,2,4-triazineoxazine compound. Applications of similar compounds. Background Technology

[0002] Agricultural pests cause crop yield losses of approximately 18-20% globally, posing a serious threat to food production and human safety. The growth and development of agricultural pests are closely related to their chitin metabolism. Chitin metabolism can be broken down into two key aspects: chitin synthesis and chitin hydrolysis. In chitin hydrolysis, chitin is first hydrolyzed into chitin oligosaccharides by family 18 glycosylhydrolases, and then further hydrolyzed into N-acetylaminohexosamine by family 20 β-N-acetylhexosamines.

[0003] Chitinases participate in the life activities of various organisms. Inhibiting chitinases in insects can lead to abnormal growth, development, molting, and death; interfering with chitinase expression in nematodes can cause growth arrest; and inhibiting fungal chitinases can affect their cell division process. Therefore, the development of novel inhibitors of chitinases is of great significance for the prevention and control of agricultural pests and diseases.

[0004] Insect chitinases of family 18 (Chi-h) are an essential part of insect molting and metamorphosis. Notably, Chi-h shares up to 70% homology with bacterial chitinases and is a unique chitinase acquired through horizontal gene transfer, exclusive to Lepidoptera. Developing novel, highly effective, and environmentally friendly insecticides targeting insect chitinases of family 18 (Chi-h) is of significant importance for the green control of agricultural pests. Summary of the Invention

[0005] The purpose of this invention is to provide a benzo-1,2,4-triazineoxazine-based... The application of similar compounds can solve the problems existing in the above-mentioned technologies and achieve effective control of lepidopteran insects.

[0006] To achieve the above objectives, the present invention provides the following solution:

[0007] One of the technical solutions of this invention: provides a benzo-1,2,4-triazineoxazine The application of this type of compound in the preparation of insecticides, wherein the insecticide is an inhibitor of insect chitinase activity.

[0008] Optionally, the insecticide targets insect species including diamondback moth and / or Asian corn borer.

[0009] Optionally, the insect chitinase comprises a glycosyl hydrolase family 18 chitinase.

[0010] Optionally, the glycosyl hydrolase family 18 chitinase comprises an Ostrinia furnacalis chitinase.

[0011] Since the insect chitinase of family 18 (Chi-h) is a chitinase obtained by gene horizontal transfer and unique to Lepidoptera insects, and considering that Lepidoptera insects are the most destructive class of crop pests, while most beneficial insects do not have Chi-h. Therefore, the present application develops a new type of inhibitor targeting Chi-h, and develops a new type of efficient green insecticide targeting insect chitinase, especially Ostrinia furnacalis chitinase (OfChi-h), which has outstanding significance for green control of agricultural pests. The present application finds that the benzene-1,2,4-triazine oxazepine compound with the structure of benzene-1,2,4-triazine oxazepine can inhibit the activity of Ostrinia furnacalis chitinase (OfChi-h) and achieve effective control of Ostrinia furnacalis and Plutella xylostella.

[0012] Preferably, the benzene-1,2,4-triazine oxazepine compound has the structural formula as shown in formula (I):

[0013]

[0014] wherein R1, R2 and R3 are independently selected from H, halogen, C1-C6 alkyl, acetyl, propionyl, butyryl, valeryl or hexanoyl;

[0015] R4 is selected from a five-membered heterocyclic group, and the five-membered heterocyclic group comprises a thiophene ring group, a furan ring group, a pyrazole ring group, a triazole ring group, a thiazole ring group or a pyrrole ring group.

[0016] The benzene-1,2,4-triazine oxazepine compound of the present application has good inhibitory effect on insect chitinase and can further achieve efficient insecticidal effect.

[0017] The benzene-1,2,4-triazine oxazepine compound has the following preparation steps:

[0018] ​The compound of formula (II) (1 equivalent) is added to a basic aqueous solution (the base used is one or two of sodium hydroxide, triethylamine, diisopropylethylamine, pyridine, potassium fluoride, sodium fluoride, sodium bicarbonate, sodium acetate, sodium carbonate, potassium carbonate, potassium acetate and ammonium acetate), a haloalkane or acid chloride (0.5-3 equivalents) is added, and the reaction is carried out in a solvent at a temperature of -20℃-100℃ (0.5-24h) to obtain a compound of formula (III);

[0019] The compound of formula (III) (1 equivalent) is added to an aldehyde compound (0.5-3 equivalents) in ethanol and glacial acetic acid, and the reaction is carried out at a temperature of 0℃-150℃ to obtain a compound of formula (IV);

[0020] The compound of formula (IV) (1 equivalent) is reacted with a haloalkane or acid chloride (0.5-3 equivalents) under basic conditions (the base used is one or two of sodium hydroxide, triethylamine, diisopropylethylamine, pyridine, potassium fluoride, sodium fluoride, sodium bicarbonate, sodium acetate, sodium carbonate, potassium carbonate, potassium acetate and ammonium acetate) in a solvent at a temperature of -20℃-100℃ to obtain a compound of formula (I).

[0021] The structural formula of the compound of formula (II), the compound of formula (III) and the compound of formula (IV) are respectively:

[0022]

[0023] wherein R1 and R2 are independently selected from H, halogen, C1-C6 alkyl, acetyl, propionyl, butyryl, valeryl or hexanoyl;

[0024] R4 is selected from a five-membered heterocyclic group, and the five-membered heterocyclic group includes a thiophene ring group, a furan ring group, a pyrazole ring group, a triazole ring group, a thiazole ring group or a pyrrole ring group.

[0025] The solvent independently includes pyridine, N,N-dimethylformamide, methanol or ethanol; and the aldehyde compound includes thiophene aldehyde, furan aldehyde, pyrazole aldehyde, triazole aldehyde, thiazole aldehyde or pyrrole aldehyde.

[0026] The second technical scheme of the present application provides an insecticide, which comprises a benzene-1,2,4-triazine oxazepine compound.

[0027] The present application discloses the following technical effects:

[0028] The benzene-1,2,4-triazine oxazepine The compound has high chitinase enzyme inhibitory activity and can exhibit excellent insecticidal effect, especially high OfChi-h chitinase inhibitory activity and strong insecticidal activity on the agricultural pests Ostrinia furnacalis and Plutella xylostella. The compound of the structure has high pesticide research value and wide application prospect in the field of pesticides. BRIEF DESCRIPTION OF DRAWINGS

[0029] Figure 1 NMR spectrum of the compound HAU-BTO12 obtained in Example 1;

[0030] Figure 2 Inhibition rate of compound HAU-BTO1 on Ostrinia furnacalis chitinase at different concentrations;

[0031] Figure 3 Inhibition rate of compound HAU-BTO2 on Ostrinia furnacalis chitinase at different concentrations;

[0032] Figure 4 Inhibition rate of compound HAU-BTO6 on Ostrinia furnacalis chitinase at different concentrations. DETAILED DESCRIPTION

[0033] The detailed description set forth below will be better understood in conjunction with the drawings as follows:

[0034] It should be understood that the terms used in the present application merely describe specific embodiments and are not intended to limit the present application. In addition, for the numerical range in the present application, it should be understood that each intermediate value between the upper limit and the lower limit of the range is also specifically disclosed. Each smaller range between any stated value or intermediate value within the stated range, and any other stated value or intermediate value within the stated range, is also included in the present application. The upper limit and the lower limit of these smaller ranges can be independently included or excluded from the range.

[0035] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, preferred methods and materials are described. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials in connection with which the documents are cited. In case of conflict, the content of the specification controls.

[0036] Many modifications and variations of the present application described in the specific embodiments of the application can be made by those skilled in the art without departing from the spirit or scope of the application. Other implementations of the application will be apparent to those skilled in the art from consideration of the specification and practice of the application disclosed herein. The specification and examples given are exemplary only.

[0037] As used herein, the terms "comprises", "comprising", "includes", "including", "has", "having", "contains", "containing", or variations thereof, are intended to be open-ended terms that mean inclusion, but not limited to, the listed material or list of materials.

[0038] The Asian corn borer chitinase (OfChi-h) involved in the following examples and application examples is obtained from the reference article "Liu, T.; Chen, L.; Zhou, Y.; Jiang, X.; Duan, Y.; Yang, Q., Structure, Catalysis, and InhibitBTOn of OfChi-h, the Lepidoptera-exclusive Insect Chitinase. J. BBTOl. Chem. 2017, 292 (6), 2080-2088."

[0039] Other raw materials are commercially available products unless otherwise specified.

[0040] Example 1

[0041] Preparation of compound HAU-BTO12, R1 is H, R2 is C2H5, R3 is CH3CO, R4 is

[0042]

[0043] (1) In a 10 mL reaction bottle, add thione compound (22.0 mg, 0.1 mmol), bromoethane (12.1 mg, 0.11 mmol), KOH (6.2 mg, 0.11 mmol), EtOH (3 mL), H2O (2 mL), stir the reaction solution at room temperature overnight, add EtOAc, separate and extract, concentrate the organic phase, and purify by column chromatography to obtain thioether compound 20.1 mg, yield 81.0%. ESI-MS: 249.26 (M+H).

[0044] (2) Dissolve thioether compound (12.4 mg, 0.05 mmol), thiophene aldehyde (6.7 mg, 0.06 mmol), glacial acetic acid (0.1 mL) in anhydrous ethanol (4 mL), reflux the reaction solution for 8 h, and purify by column chromatography to obtain oxazoline compound 10.2 mg, yield 73.0%. ESI-MS: 249.26 (M+H). Compound 8.2 mg, yield 47.9%. ESI-MS: 343.58 (M+H).

[0045] (3) The oxygen-nitrogen heterocycle Compound (17.1 mg, 0.05 mmol), acetyl chloride (4.7 mg, 0.06 mmol) was dissolved in anhydrous pyridine, and the reaction was carried out at room temperature overnight. The reaction solution was concentrated and purified by column chromatography to obtain compound 16.8 mg of HAU-BTO12, yield 87.5%.

[0046] The structural confirmation data of compound HAU-BTO12 are as follows: 1 H NMR (400 MHz, DMSO-d6) δ 8.92 (d, 1H), 7.78-7.24 (m, 6H), 6.99-6.89 (m, 1H), 3.42-3.15 (m, 2H), 2.31 (s, 3H), 1.38 (t, 3H); HRMS (ESI) calcd for C 18 H 17 N4O2S2(M+H + ) 385.0793 found 385.0799.

[0047] Figure 1 The nuclear magnetic hydrogen spectrum (400 MHz, DMSO-d6) of the compound HAU-BTO12 obtained in the present example.

[0048] Example 2

[0049] Using different substituent raw materials, the preparation method described in Example 1 was used to prepare compounds HAU-BTO1, HAU-BTO2, HAU-BTO3, HAU-BTO4, HAU-BTO5, HAU-BTO6, HAU-BTO7, HAU-BTO8, HAU-BTO9, HAU-BTO10, HAU-BTO11, HAU-BTO13, HAU-BTO14, HAU-BTO15, HAU-BTO16, HAU-BTO17, HAU-BTO18, HAU-BTO19 and HAU-BTO20, respectively.

[0050] Application Example 1

[0051] The inhibition rate of the above obtained benzene-1, 2, 4-triazine and oxygen-nitrogen heterocycle compounds on the chitinase (OfChi-h) of Asian corn borer was tested:

[0052] Test method:

[0053] Experimental group: (each compound concentration of 10 μM experiment) set 3 groups of parallel experimental group. In 30 ℃ reaction temperature, 100 μL of the reaction system was added 2 nmol / L of Ostrinia furnacalis chitinase, 10 μM of each compound and 50 μM of substrate 4-Mu-(GlcNAc)2 in 20 mM of pH=6.0 phosphate buffer solution was incubated for 30 min; 0.5 M sodium carbonate solution was added to terminate the reaction, the reaction solution was excited by 360 nm wavelength excitation light, and the absorbance value at 450 nm wavelength was measured.

[0054] (5 μM concentration of each compound experiment) set 3 groups of parallel experimental group. In 30 ℃ reaction temperature, 100 μL of the reaction system was added 2 nmol / L of Ostrinia furnacalis chitinase, 5 μM of each compound and 50 μM of substrate 4-Mu-(GlcNAc)2 in 20 mM of pH=6.0 phosphate buffer solution was incubated for 30 min; 0.5 M sodium carbonate solution was added to terminate the reaction, the reaction solution was excited by 360 nm wavelength excitation light, and the absorbance value at 450 nm wavelength was measured.

[0055] Positive control: set 3 groups of parallel positive control. In 30 ℃ reaction temperature, 100 μL of the reaction system was added 2 nmol / L of Ostrinia furnacalis chitinase, 50 μM of substrate 4-Mu-(GlcNAc)2 in 20 mM of pH=6.0 phosphate buffer solution was incubated for 30 min; 0.5 M sodium carbonate solution was added to terminate the reaction, the reaction solution was excited by 360 nm wavelength excitation light, and the absorbance value at 450 nm wavelength was measured.

[0056] The enzyme inhibition activity of each compound was calculated according to the following formula: inhibition percentage=(absorbance of positive control-absorbance of experimental group) / absorbance of positive control*100%.

[0057] The results are shown in Table 1.

[0058] Table 1 enzyme inhibition activity of each compound

[0059]

[0060]

[0061]

[0062]

[0063]

[0064] As shown in Table 1, the benzene-1,2,4-triazine and oxazepine These compounds exhibited excellent inhibitory activity against chitinase (OfChi-h) in Asian corn borer. Among them, compounds HAU-BTO1, HAU-BTO2, HAU-BTO4, HAU-BTO6, and HAU-BTO20 all showed inhibition rates greater than 75% against OfChi-h at a concentration of 10 μM. Compounds HAU-BTO1, HAU-BTO2, and HAU-BTO6 showed high levels of inhibition against OfChi-h activity, with inhibition percentages of 88.6–93.8% at 10 μM and 72.5–87.6% at 5 μM.

[0065] Application Example 2

[0066] The half-maximal inhibitory concentration (IC50) of compounds HAU-BTO1, HAU-BTO2, and HAU-BTO6 against chitinase (OfChi-h) in Asian maize borer was tested. 50 value:

[0067] Test method: At a reaction temperature of 30℃, 2 nmol / L Asian corn borer chitinase, a certain concentration gradient of compounds (HAU-BTO1, HAU-BTO2, or HAU-BTO6) (with a logarithmic variation range of -0.5 to 1 μM), and 50 μM substrate 4-Mu-(GlcNAc)2 were added to a 100 μL reaction system and incubated in 20 mM phosphate buffer (pH 6.0) for 30 min. The reaction was terminated by adding 0.5 M sodium carbonate solution. The reaction solution was excited with excitation light at a wavelength of 360 nm, and the absorbance was measured at a wavelength of 450 nm. Data were processed and plotted using Graphpad Prism. The results are shown below. Figures 2 to 4 As shown.

[0068] Figure 2 The inhibition rate of different concentrations of the compound HAU-BTO1 on chitinase in Asian corn borer was measured. Figure 3 The inhibition rate of different concentrations of compound HAU-BTO2 on chitinase in Asian corn borer was measured. Figure 4 The inhibition rate of different concentrations of the compound HAU-BTO6 on chitinase in Asian corn borer was shown. Figures 2 to 4 The horizontal axis represents the logarithm of the concentration of each compound, and the vertical axis represents the percentage inhibition rate of each compound on chitinase in Asian corn borer.

[0069] Depend on Figures 2 to 4 It was found that compounds HAU-BTO1, HAU-BTO2, and HAU-BTO6 all exhibited strong inhibitory effects on the chitinase activity of Asian corn borer at relatively low concentrations. Among them, compound HAU-BTO1 had the highest IC50 value. 50 =1.60 μM, IC50 of compound HAU-BTO250 = 2.36 μM, IC50of compound HAU-BTO6 50 = 2.41 μM.

[0070] Example 3

[0071] The insecticidal activity of the benzo-1,2,4-triazine and oxazepine compounds obtained in the above examples was tested.

[0072] The insecticidal activity of the benzo-1,2,4-triazine and oxazepine compounds obtained in the above examples was tested. The insecticidal activity of the benzo-1,2,4-triazine and oxazepine compounds obtained in the above examples was tested.

[0073] The corrected mortality rate was calculated according to the following formula: corrected mortality rate (%) = (experimental group mortality rate - blank control mortality rate) / (1 - blank control mortality rate) * 100%.

[0074] The results are shown in Table 2.

[0075] Table 2 Insecticidal effect of the compounds obtained in each example

[0076]

[0077]

[0078] As can be seen from Table 2, the benzo-1,2,4-triazine and oxazepine compounds of the present application have excellent insecticidal effect; the insecticidal activity of most of the compounds against Plutella xylostella and Ostrinia furnacalis larvae at a concentration of 500 μg / mL is 100%; the insecticidal activity of compounds HAU-BTO1, HAU-BTO2 and HAU-BTO6 against Plutella xylostella larvae at a concentration of 200 μg / mL is not less than 70%, which is superior to the commercially available flufenoxuron; the insecticidal activity of compounds HAU-BTO1, HAU-BTO2 and HAU-BTO6 against Ostrinia furnacalis larvae at a concentration of 200 μg / mL is not less than 80%, which is superior to the commercially available flufenoxuron.

[0079] Each example in the present specification is described in a progressive manner, and each example focuses on the differences from other examples. The same or similar parts between each example can be referred to each other.​

[0080] The foregoing description of the embodiments disclosed will so fully reveal the general nature of the application that others can, by applying current knowledge of the art, adapt it for other purposes or modify it to suits the particular specifications or testing environment to which it is applied, but such departures are deemed within the spirit and scope of the application. Accordingly, the application is not limited as exemplified herein.

Claims

1. The application of a benzo-1,2,4-triazinoxazine compound in the preparation of an insect chitinase activity inhibitor, characterized in that, The benzene-1,2,4-triazinooxazepine compound has a structural formula as shown in formula (I): Formula (I); In the formula, R1 is H or halogen; R2 is C1-C6 alkyl; R3 is H, acetyl, propionyl or butyryl; and R4 is a thiazole ring group, an imidazole ring group, a pyrrole ring group, a furan ring group or a thiophene ring group.

2. Use according to claim 1, characterized in that, The insect species targeted include Plutella xylostella and / or Ostrinia nubilalis.

3. Use according to claim 1, characterized in that, The insect chitinases include glycosyl hydrolase family 18 chitinases.

4. Use according to claim 3, characterized in that, The glycosyl hydrolase family 18 chitinases include Ostrinia nubilalis chitinases.

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