Application of benzo-1, 2, 4-triazine oxygen aza compound

Abstract

The invention discloses an application of a benzo-1, 2, 4-triazine oxygen aza # imgabs0 # compound, and belongs to the technical field of insecticide preparation. The benzo-1, 2, 4-triazine oxygen aza # imgabs 1 # compound is used for preparing an insecticide, and the compound has relatively high enzyme inhibition activity on insect chitinase, can show an excellent insecticidal effect, and particularly has relatively high inhibition activity on Asiatic corn borer chitinase (OfChii-h); the insecticidal composition has strong insecticidal activity on agricultural pests such as ostrinia furnacalis and plutella xylostella. The compound with the structure is used for preparing insecticides, has very high pesticide research value, and has wide application prospects in the field of pesticide science.

Description

Technical Field

[0001] The present invention belongs to the technical field of insecticide preparation, and more specifically relates to a benzo-1,2,4-triazine oxazepine Application of compounds. Background Art

[0002] Globally, agricultural pests cause crop yield losses of about 18-20%, and agricultural pests seriously threaten food production and human safety. The growth and development of agricultural pests are closely related to their chitin metabolism. The metabolic process of chitin can be divided into two key aspects: one is chitin anabolism; the other is chitin hydrolysis. In the chitin hydrolysis process, chitin is first hydrolyzed into chito-oligosaccharides by chitinase of family 18 of glycosyl hydrolase, and then hydrolyzed into N-acetylhexosamine by β-N-acetylhexosaminidase of family 20.

[0003] Chitinase can participate in the life activities of many organisms. Inhibiting insect chitinase will cause insects to fail to grow and develop normally and molt and die. Interfering with the expression of chitinase in nematodes will cause the growth of nematodes to stagnate. Inhibiting fungal chitinase will affect its division process. Therefore, the discovery of new inhibitors for chitinase is of great significance for the prevention and control of agricultural pests and diseases.

[0004] Insect chitinase (Chi-h) of family 18 is an indispensable part of insect molting and metamorphosis. It is worth mentioning that Chi-h has up to 70% homology with chitinase from bacteria and is a chitinase unique to Lepidoptera insects obtained through horizontal gene transfer. The development of new and highly effective green insecticides targeting insect chitinase (Chi-h) of family 18 is of outstanding significance for the green control of agricultural pests. Summary of the invention

[0005] The purpose of the present invention is to provide a benzo-1,2,4-triazine oxazepine The application of such compounds can solve the problems existing in the above-mentioned prior art and achieve effective control of Lepidoptera insects.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] One of the technical solutions of the present invention is to provide a benzo-1,2,4-triazine oxazepine The invention discloses an application of the compound in the preparation of insecticides, wherein the insecticides are insect chitinase activity inhibitors.

[0008] Optionally, the insect species targeted by the pesticide include Plutella xylostella 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 includes Ostrinia furnacalis chitinase.

[0011] Since the 18-family insect chitinase (Chi-h) is a chitinase unique to Lepidoptera insects that is obtained through horizontal gene transfer. Considering that Lepidoptera insects are the most destructive type of crop pests, and most beneficial insects do not have Chi-h. Therefore, the present invention carries out research and development of new inhibitors targeting Chi-h, targeting insect chitinase, especially Asian corn borer chitinase (OfChi-h), and developing new and efficient green pesticides, which has outstanding significance for the green prevention and control of agricultural pests. The present invention found that benzo-1,2,4-triazine oxazepine The structure of benzo-1,2,4-triazine oxazepine The compounds are mainly benzo-1,2,4-triazine-oxazepine The presence of the structure can inhibit the activity of Asian corn borer chitinase (OfChi-h), thereby achieving effective prevention and control of Asian corn borer and diamondback moth.

[0012] Preferably, the benzo-1,2,4-triazine oxazepine The structural formula of the compound is 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 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.

[0016] The benzo-1,2,4-triazine oxazepine of the present invention Such compounds have a good inhibitory effect on insect chitinase and can further achieve a highly efficient insecticidal effect.

[0017] The benzo-1,2,4-triazine oxazepine The preparation steps of the compounds are as follows:

[0018] The compound of formula (II) (1 equivalent) is added to an alkaline 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), and a halogenated alkane or an acid chloride (0.5 to 3 equivalents) is added, and the reaction is carried out in a solvent at a temperature of -20°C to 100°C (0.5 to 24 hours) to obtain a compound represented by formula (III);

[0019] Adding the compound represented by formula (III) (1 equivalent) and the aldehyde compound (0.5 to 3 equivalents) to ethanol and glacial acetic acid, and reacting at a temperature of 0°C to 150°C to obtain the compound represented by formula (IV);

[0020] The compound represented by formula (IV) (1 equivalent) is reacted with a halogenated alkane or an acyl chloride (0.5 to 3 equivalents) under alkaline 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°C to 100°C to obtain a compound represented by formula (I).

[0021] The structural formulas 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; the aldehyde compound includes thiophene aldehyde, furan aldehyde, pyrazole aldehyde, triazole aldehyde, thiazole aldehyde or pyrrole aldehyde.

[0026] The second technical solution of the present invention is to provide an insecticide, the insecticide comprising benzo-1,2,4-triazine oxazepine Class compounds.

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

[0028] The benzo-1,2,4-triazine oxazepine having the structure of formula (I) of the present invention The compounds have high enzyme inhibition activity against insect chitinase and can show excellent insecticidal effect, especially high inhibition activity against Asian corn borer chitinase (OfChi-h), and strong insecticidal activity against agricultural pests such as Asian corn borer and diamondback moth. The compounds with this structure are used in the preparation of insecticides, have high pesticide research value, and have broad application prospects in the field of pesticide science. BRIEF DESCRIPTION OF THE DRAWINGS

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

[0030] Figure 2 is the inhibition rate of different concentrations of compound HAU-BTO1 on chitinase of Asian corn borer;

[0031] Figure 3 is the inhibition rate of different concentrations of compound HAU-BTO2 on chitinase of Asian corn borer;

[0032] Figure 4 The inhibition rate of different concentrations of compound HAU-BTO6 on chitinase of Asian corn borer. DETAILED DESCRIPTION

[0033] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0034] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0035] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0036] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0037] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

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

[0039] Other raw materials are commercially available 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) Add thioketone compound into a 10 mL reaction bottle. (22.0 mg, 0.1 mmol), ethyl bromide (12.1 mg, 0.11 mmol), KOH (6.2 mg, 0.11 mmol), EtOH (3 mL), H2O (2 mL), the reaction solution was stirred at room temperature overnight, EtOAc was added, the liquids were separated and extracted, the organic phase was concentrated, and column chromatography was used for purification to obtain 20.1 mg of sulfide compound, with a yield of 81.0%. ESI-MS: 249.26 (M+H).

[0044] (2) The sulfide compound (12.4 mg, 0.05 mmol), thiophene aldehyde (6.7 mg, 0.06 mmol), and glacial acetic acid (0.1 mL) were dissolved in anhydrous ethanol (4 mL), and the reaction solution was refluxed for 8 h. After purification by column chromatography, oxazapine was obtained. Compound 8.2 mg, yield 47.9%. ESI-MS: 343.58 (M+H).

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

[0046] The structural confirmation data of compound HAU-BTO12 are as follows: 1 H NMR(400MHz,DMSO-d6)δ8.92(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 + )385.0793found 385.0799.

[0047] Figure 1 This is the H NMR spectrum (400 MHz, DMSO-d6) of the compound HAU-BTO12 obtained in this example.

[0048] Example 2

[0049] By referring to the preparation method described in Example 1, using raw materials with different substituents, 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 were prepared respectively.

[0050] Application Example 1

[0051] Test the benzo-1,2,4-triazine oxazepine obtained in the above example Inhibition rate of compounds on chitinase (OfChi-h) of Asian corn borer:

[0052] Test method:

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

[0054] (Experiments with a 5 μM concentration of the compound obtained in each example) Set up 3 parallel experimental groups. At a reaction temperature of 30°C, 2 nmol / L of Ostrinia furnacalis chitinase, 5 μM of the compound obtained in each example, and 50 μM of substrate 4-Mu-(GlcNAc)2 were added to a 100 μL reaction system and incubated for 30 min in a 20 mM phosphate buffer solution at pH = 6.0; 0.5 M sodium carbonate solution was added to terminate the reaction, and the reaction solution was excited with an excitation light of 360 nm wavelength and then the absorbance value at a wavelength of 450 nm was measured.

[0055] Positive control: 3 sets of parallel positive controls were set up. At a reaction temperature of 30°C, 2nmol / L Asian corn borer chitinase and 50μM substrate 4-Mu-(GlcNAc)2 were added to 100μL of the reaction system and incubated in 20mM pH=6.0 phosphate buffer for 30min; 0.5M sodium carbonate solution was added to terminate the reaction, and the reaction solution was excited with 360nm wavelength excitation light and the absorbance value at 450nm wavelength was measured.

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

[0057] The results are shown in Table 1.

[0058] Table 1 Enzyme inhibitory activity of the compounds obtained in each example

[0059]

[0060]

[0061]

[0062]

[0063]

[0064] As shown in Table 1, the benzo-1,2,4-triazine oxazepine The compounds showed excellent inhibitory effects on the activity of Asian corn borer chitinase (OfChi-h), among which the inhibition rates of compounds HAU-BTO1, HAU-BTO2, HAU-BTO4, HAU-BTO6 and HAU-BTO20 on OfChi-h were all greater than 75% at a concentration of 10 μM; the inhibitory effects of compounds HAU-BTO1, HAU-BTO2 and HAU-BTO6 on the activity of OfChi-h reached a relatively high level, with the inhibition percentages at a concentration of 10 μM reaching 88.6 to 93.8%, and the inhibition percentages at a concentration of 5 μM reaching 72.5 to 87.6%.

[0065] Application Example 2

[0066] IC50 of test compounds HAU-BTO1, HAU-BTO2 and HAU-BTO6 against chitinase (OfChi-h) of Asian corn borer 50 value:

[0067] Test method: At a reaction temperature of 30°C, 2nmol / L Asian corn borer chitinase, a certain concentration gradient of compounds (HAU-BTO1, HAU-BTO2 or HAU-BTO6) (the logarithm of the concentration ranges from -0.5 to 1μM) and 50μM substrate 4-Mu-(GlcNAc)2 were added to a 100μL reaction system and incubated in a 20mM pH=6.0 phosphate buffer for 30min; 0.5M sodium carbonate solution was added to terminate the reaction, and the reaction solution was excited with 360nm wavelength excitation light and the absorbance value at 450nm wavelength was measured. Graphpad Prism was used to process the data and plot the results. Figures 2 to 4 shown.

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

[0069] Depend on Figures 2 to 4 It can be seen that compounds HAU-BTO1, HAU-BTO2 and HAU-BTO6 can strongly inhibit the activity of chitinase in Asian corn borer at lower concentrations. Among them, the IC 50 =1.60μM, IC of compound HAU-BTO250 =2.36μM, IC of compound HAU-BTO6 50 =2.41μM.

[0070] Application Example 3

[0071] Test the benzo-1,2,4-triazine oxazepine obtained in the above example Insecticidal activity of compounds:

[0072] Test method: The benzo-1,2,4-triazine oxazepine obtained in the above example was tested by leaf dipping method. The insecticidal activity of the compounds against the larvae of diamondback moth and Asian corn borer was studied. The young corn leaves were immersed in 500μg / mL and 200μg / mL of each compound for 10s, and then naturally dried. The medicated corn leaves were then fed to the third-instar larvae of diamondback moth and Asian corn borer. The results were checked every 24h and the mortality rate was calculated. The commercially available hexaflumuron was selected as the positive control. (The results obtained by the treatment of each compound and hexaflumuron were recorded as the mortality rate of the experimental group). A blank control (recorded as CK) was set, and no reagent was added to the blank control.

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

[0074] The results are shown in Table 2.

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

[0076]

[0077]

[0078] As can be seen from Table 2, the benzo-1,2,4-triazine oxazepine #imgpt32# compounds of the present invention have excellent insecticidal effects; the insecticidal activity of most of the compounds against the larvae of diamondback moth and the larvae of Asian corn borer at a concentration of 500 μg / mL is 100%; the insecticidal activity of the compounds HAU-BTO1, HAU-BTO2, and HAU-BTO6 against the larvae of diamondback moth at a concentration of 200 μg / mL is not less than 70%, which is better than the commercially available hexaflumuron; the insecticidal activity of the compounds HAU-BTO1, HAU-BTO2, and HAU-BTO6 against the larvae of Asian corn borer at a concentration of 200 μg / mL is not less than 80%, which is better than the commercially available hexaflumuron.

[0079] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0080] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A benzo-1,2,4-triazine oxazepine The use of a compound in the preparation of an insecticide is characterized in that: The insecticide is an insect chitinase activity inhibitor.

2. The use according to claim 1, characterized in that: The insect species targeted by the pesticide include diamondback moth and / or Asian corn borer.

3. The use according to claim 1, characterized in that: The insect chitinase includes glycosyl hydrolase 18 family chitinase.

4. The use according to claim 3, characterized in that: The glycosyl hydrolase 18 family chitinase includes Ostrinia furnacalis chitinase.

5. The use according to claim 1, characterized in that: The benzo-1,2,4-triazine oxazepine The structural formula of the compound is shown in formula (I): wherein R1, R2 and R3 are independently selected from H, halogen, C1-C6 alkyl, acetyl, propionyl, butyryl, valeryl or hexanoyl; 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.

6. An insecticide, characterized in that: The pesticide includes benzo-1,2,4-triazine oxazepine Class compounds.

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