A triazoloquinazolinone compound and its application

Abstract

The invention discloses a triazoloquinazolinone compound and its application, and belongs to the technical field of pesticides. The triazoloquinazolinone compound has excellent inhibitory activity and excellent insecticidal activity on chitinase, especially has high inhibitory activity on chitinase h of Asian corn borer; and has strong insecticidal activity on agricultural pests corn borer and diamondback moth. The compound can be used as chitinase inhibitor or insecticide, has high pesticide research value, and has broad application prospects in the field of pesticide science.

Description

Technical Field

[0001] The invention belongs to the technical field of pesticides, and specifically relates to a triazoloquinazolinone compound and application thereof. Background Art

[0002] Chitin, also known as chitosan, is a type of polysaccharide compound widely found in nature. Chitin is composed of N-acetylglucosamine as a monomer connected by β-1,4 glycosidic bonds. It exists in the shells of crustaceans such as shrimp, crabs, and insects, the cell walls of fungi, and the egg shells of nematodes.

[0003] The hydrolysis and metabolism of chitin is mainly completed by chitinase and β-N-acetylhexosaminidase. The chitinase of family 18 is responsible for hydrolyzing chitin into chito-oligosaccharides, and the β-N-acetylhexosaminidase of family 20 hydrolyzes chito-oligosaccharides into single N-acetylglucosamine. The hydrolysis and metabolism of chitin plays a vital role in the growth and development of insects, nematodes, and fungi. For example, inhibiting the chitinase of insects will cause the insects to die due to the inability to grow and develop normally, molting, and inhibiting the activity of fungal chitinase will affect the division process of fungi. In addition, since chitin is not contained in humans and higher animals and plants, chitinase that catalyzes the degradation of chitin is recognized as an important target for green pesticides.

[0004] Several types of chitinase inhibitors have been reported, including chitosan, rhodanine, TMG-(GlcNAc)4, phlegmacinB1, polonimidesA-C, berberine, thiazole acylhydrazone, bipyridyl pyrimidine, azopyrimidine, aza macrolide, etc. Most of these inhibitor molecules have problems such as difficulty in sourcing and poor drugability.

[0005] Therefore, it is of great significance for the prevention and control of agricultural diseases and insect pests to develop new chitinase inhibitors and obtain chitinase inhibitor molecules with novel skeletons and good drug properties. Summary of the invention

[0006] The purpose of the present invention is to provide a triazoloquinazolinone compound and its application. The triazoloquinazolinone compound is as shown in formula (I), and the structural compound has excellent inhibitory activity on chitinase and excellent insecticidal activity, can be used as a chitinase inhibitor or insecticide, has high pesticide research value, and has broad application prospects in the field of pesticide science.

[0007] To achieve the above object, the present invention provides the following technical solutions:

[0008] One of the technical solutions of the present invention is to provide a triazoloquinazolinone compound, the structural formula of which is shown in formula (I):

[0009]

[0010] In formula (I), R1 is selected from H, C1-C8 alkyl, halogen, OH, NH2, alkyl carboxylic acid or alkyl amide;

[0011] R2 is selected from H, C1-C8 alkyl, substituted phenyl, C1-C8 alkoxy, Wherein m=1-8, n=1-8;

[0012] R3-R6 are independently selected from H, C1-C4 alkyl, halogen, OCH3, CN, NHAc or NO2.

[0013] Preferably, said R1 is Wherein, R1′ is selected from H, C1-C4 alkyl, substituted phenyl or substituted benzyl.

[0014] More preferably, the R1′ is selected from

[0015] Preferably, said R2 is

[0016] The second technical solution of the present invention is to provide a method for preparing the above-mentioned triazoloquinazolinone compound, comprising the following steps:

[0017] A quinazolinone compound, potassium carbonate, anhydrous ethanol and hydrazine hydrate are mixed to obtain a hydrazine quinazolinone compound after reaction; the hydrazine quinazolinone compound and a carboxyl compound are dissolved in toluene to obtain a triazoloquinazolinone compound after reaction;

[0018] The structural formula of the quinazolinone compound is shown in formula (II):

[0019]

[0020] The structural formula of the hydrazine quinazolinone compound is shown in formula (III):

[0021]

[0022] The structural formula of the carboxyl compound is shown in formula (IV):

[0023] R1-COOH

[0024] Formula (IV).

[0025] In formula (II) to formula (IV), the selection of R1 to R6 refers to R1 to R6 of the aforementioned triazoloquinazolinone compounds.

[0026] Preferably, the molar ratio of the quinazolinone compound, potassium carbonate and hydrazine hydrate is 1:0.5-3:0.1-10; the molar ratio of the hydrazine quinazolinone compound to the carboxyl compound is 1:10-1000.

[0027] The reaction conditions of the hydrazine quinazolinone compound are reflux reaction at 30-80° C. for 5-48 hours; the reaction conditions of the triazoloquinazolinone compound are reflux reaction at 80-180° C. for 0.5-72 hours.

[0028] Technical solution three of the present invention: Use of the triazoloquinazolinone compound described in any one of claims 1 to 4 in the preparation of drugs for inhibiting chitinase activity or in the preparation of drugs for controlling agricultural pests.

[0029] Preferably, the chitinase is a glycosyl hydrolase family 18 chitinase.

[0030] Preferably, the chitinase of glycosyl hydrolase family 18 is Ostrinia furnacalis chitinase H. When inhibiting Ostrinia furnacalis chitinase H, the required concentration of the triazoloquinazolinone compound is not less than 0.1 μM.

[0031] Preferably, the agricultural pests include Ostrinia nubilalis or Plutella xylostella.

[0032] The beneficial technical effects of the present invention are as follows:

[0033] The present invention finds that the triazoloquinazolinone compound with the structure of formula (I) has excellent inhibitory activity and excellent insecticidal activity on chitinase, especially has high inhibitory activity on Asian corn borer chitinase h (OfChi-h); and has strong insecticidal activity on agricultural pests corn borer and diamondback moth. The compound with this structure can be used as chitinase inhibitor or insecticide, has high pesticide research value, and has broad application prospects in the field of pesticide science. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 This is the H NMR spectrum (400 MHz, DMSO) of compound DB-2.

[0035] Figure 2 IC of compound DB20 on OfChi-h 50 value.

[0036] Figure 3 IC of compound DB22 against OfChi-h 50 value. DETAILED DESCRIPTION

[0037] Now, various exemplary embodiments of the present invention are described in detail, and this detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics and embodiments of the present invention. It should be understood that the terms described in the present invention are only for describing specific embodiments and are not used to limit the present invention.

[0038] In addition, for the numerical range in the present invention, it is understood that each intermediate value between the upper and lower limits of the range 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 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.

[0039] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the invention pertains. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention.

[0040] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0041] The chitinase OfChi-h in the embodiment of the present invention is given in the following references:

[0042] Liu, T.; Chen, L.; Zhou, Y.; Jiang,

[0043] Examples 1-2 of the present invention disclose the synthesis methods of representative compounds DB-1 to DB-26. The specific structures of DB-1 to DB-26 are shown in Table 1.

[0044] Taking compound DB-2 as an example, the preparation and identification of the compound of the present invention are described.

[0045] Example 1

[0046] Compound DB-2( The synthesis of R2=isobutyl, R3=R4=R5=R6=H) is as follows:

[0047]

[0048] The specific preparation steps are:

[0049] (1) In a 50 mL flask, a quinazolinone compound (234 mg, 1 mmol), potassium carbonate (138 mg, 1 mmol) and anhydrous ethanol (10 mL) were added, and hydrazine hydrate (625 mg, 10 mmol) was added. The reaction system was refluxed at 78 ° C for 24 h, cooled to room temperature, poured into ice water, filtered, concentrated, and recrystallized from ethanol to obtain 174 mg of a hydrazine quinazolinone compound with a yield of 75%. ESI-MS: 233.56 (M+H).

[0050] (2) A hydrazinoquinazolinone compound (100 mg, 0.43 mmol) and a carboxyl compound (89 mg, 0.43 mmol) were dissolved in toluene and reacted at 110° C. under reflux for 48 h. The reaction solution was concentrated in vacuo and purified by column chromatography (DCM / MeOH, 30:1 v / v) to obtain 55.4 mg of the target compound with a yield of 32%.

[0051] The structural confirmation data are as follows: 1 HNMR(400MHz,DMSO)δ8.52(t,J=8.0Hz,1H,NH),8.20–8.29(m,1H,ArH),8.04–8.08 (m,1H,ArH),7.98–7.89(m,1H,ArH),7.64–7.53(m,1H,ArH),7.37–7.16(m,5H,ArH) ,4.45–4.40(m,2H,CH2),4.09–4.01(m,2H,CH2),3.58–3.41(m,2H,CH2),2.81–2.7 3(m,2H,CH2),2.41–2.20(m,1H,CH),1.10–0.94(m,6H,2CH3); HRMS(ESI)calcdforC 23 H 26 N5O2(M+H + )404.2087,found404.2091.

[0052] Figure 1 This is the H NMR spectrum (400 MHz, DMSO) of compound DB-2.

[0053] Example 2

[0054] The synthesis of compound DB-1 and compounds DB-3 to DB-26 was carried out in accordance with Example 1, except that the starting materials, quinazolinone compound and carboxyl compound, were selected differently.

[0055] Example 3

[0056] The inhibition rate of compounds DB-1 to DB-26 on chitinase OfChi-h was determined by the following specific steps:

[0057] Experimental group: 3 parallel experimental groups were set up. At a reaction temperature of 30°C, 2nmol / L chitinase, 10μM compound (or 1μM compound) and 50μM substrate 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 at 450nm wavelength was measured.

[0058] Positive control: 3 sets of parallel positive controls were set up. At 30°C, 2 nmol / L chitinase and 50 μM substrate Mu-GlcNAc were added to 100 μL of the reaction system and incubated in 20 mM pH 6.0 phosphate buffer for 30 min; 0.5 M sodium carbonate solution was added to terminate the reaction, and the reaction solution was excited with 360 nm wavelength excitation light and the absorbance at 450 nm wavelength was measured.

[0059] The enzyme inhibitory activity of the compounds was calculated according to the following formula:

[0060] Inhibition percentage = (positive control - experimental group) / positive control × 100

[0061] The results are shown in Table 1, which lists the numbers, chemical structures and enzyme inhibitory activities of 26 representative compounds against OfChi-h.

[0062] Table 1

[0063]

[0064]

[0065]

[0066]

[0067] Example 4

[0068] IC50 of compounds DB-1 to DB-26 against OfChi-h 50 The specific steps for determining the value are as follows:

[0069] At 30°C, 2nmol / L chitinase, compounds with appropriate concentration gradients, and 50μM substrate 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 at 450nm wavelength was measured. GraphpadPrism was used to process the data and draw the graph. The IC values ​​of compounds DB-5, DB-7, DB-20, and DB-22 against OfChi-h in Table 1 are shown in Table 1. 50 The values ​​were 1.152μM, 0.799μM, 1.333μM, and 0.670μM, respectively. Half inhibition concentration IC 50 The values ​​showed that the triazoloquinazolinone compounds DB-5, DB-7, DB-20, and DB-22 had excellent inhibitory activity against OfChi-h. Figure 2 IC of compound DB20 on OfChi-h 50 value. Figure 3 IC of compound DB22 against OfChi-h 50 value.

[0070] Example 5

[0071] Insecticidal activity assay of compounds DB-1 to DB-26:

[0072] The insecticidal activity of triazoloquinazolinone compounds against corn borer and diamondback moth was tested by insect immersion method. The third-instar corn borer and diamondback moth larvae were immersed in the compound solution with a concentration of 500μg / mL for 5-10 seconds, and the excess solution was absorbed with filter paper. Then the test insects were transferred to normal conditions for 48 hours, the results were checked, and the mortality rate was calculated. Hexaflumuron was selected as the positive control. The formula for correcting the mortality rate is as follows, and the calculation results are shown in Table 2.

[0073] Corrected mortality rate (%) = (sample mortality rate - blank control mortality rate) / (1 - blank control mortality rate) × 100.

[0074] Table 2 Insecticidal activity test results of compounds DB-1 to DB-26

[0075]

[0076]

[0077] It can be seen from the above experiments that the triazoloquinazolinone compounds represented by the structure of formula (I) of the present invention have excellent enzyme inhibitory activity against chitinase, and the inhibitory activity data on chitinase (including the inhibition rate of the inhibitor, the half inhibition concentration IC 50) showed that the 26 representative triazoloquinazolinone compounds shown in Table 1 exhibited inhibitory activity against chitinase OfChi-h. In particular, the inhibition rates of compounds DB-3, DB-5, DB-6, DB-7, DB-8, DB-12, DB-13, DB-14, DB-20, and DB-21 in Table 1 were all greater than 70% at a test concentration of 10 μM against OfChi-h; the inhibitory activities of compounds DB-5, DB-6, DB-7, DB-9, DB-20, and DB-22 in Table 1 against OfChi-h were all greater than 50% at a test concentration of 1 μM. The IC values ​​of compounds DB-5, DB-6, DB-7, DB-9, DB-20, and DB-22 in Table 1 against OfChi-h were 50 The values ​​are 1.152 μM, 2.100 μM, 4.324 μM, 0.799 μM, 1.333 μM, and 0.670 μM, respectively.

[0078] In addition, the compounds shown in the structural formula (I) have strong insecticidal activity against agricultural pests. The insecticidal activity of compounds DB-1, DB-4, DB-6, DB-8, DB-9, DB-10, and DB-18 in Table 2 against corn borers is greater than 50%, which is better than the control drug hexaflumuron; the insecticidal activity of compounds DB-1, DB-3, DB-6, DB-14, DB-25, and DB-26 in Table 1 against diamondback moth at a concentration of 500 μg / mL is 100%, which is much better than the control drug hexaflumuron. The results all show that the compounds shown in the structural formula (I) have good application prospects in the field of plant protection.

[0079] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Use of triazoloquinazolinone compounds in the preparation of drugs for inhibiting chitinase activity or drugs for preventing and controlling corn borers or diamondback moths, characterized in that: The structural formula of the triazoloquinazolinone compound is shown in formula (I): In formula (I), R1 is Wherein, R1′ is selected from R2 is R3-R6 are each independently selected from H.

2. The use according to claim 1, characterized in that: The chitinase is a glycosyl hydrolase 18 family chitinase.

3. The use according to claim 2, characterized in that: The glycosyl hydrolase 18 family chitinase is Ostrinia nubilalis chitinase h.

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