RELATED APPLICATIONS

The present application is a U.S. National Phase of International Application Number PCT/CN2021/105643 filed Jul. 12, 2021, and claims priority to Chinese Application Number 202110005578.6, filed Jan. 5, 2021, and Chinese Application Number 202110760647.4 filed Jul. 6, 2021.

TECHNICAL FIELD

The present invention relates to the technical field of insecticides, in particular to diphenylpyrazole-based compounds, the preparation method therefor and use thereof.

BACKGROUND ART

Insecticides have long been the primary means of agricultural pest control. Among many insecticides, the ryanodine receptor modulator represented by chlorantraniliprole and the voltage-dependent sodium ion channel blocker represented by indoxacarb have novel structure, unique mechanism of action, quick effect, wide insecticidal spectrum, and environmental friendliness. Thus they have received widespread attention and become a popular variety of insecticides. Chlorantraniliprole belongs to the diamide-based insecticides, and indoxacarb belongs to the oxadiazine-based insecticides. Both types of insecticides can control most pests with chewing mouthparts, particularly, they have good control effects on Pyralidae (Cnaphalocrocis medinalis, Pyrausta nubilalis, Maruca vitrata Fabricius, etc.), Carposinidae (Carposina sasakii, etc.), Noctuidae (Spodoptera frugiperda, Helicoverpa armigera, Spodoptera litura, etc.), Tortricidae (Cydia pomonella, Grapholita molesta, etc.), Gelechiidae (Pectinophora gossypiella, etc.), Plutellidae (Plutella xylostella etc.), Pieridae (Pieris rapae, etc.), Gracilariidae (Lithocolletis ringoniella Mats., etc.), etc. of Lepidoptera, and they also have high activity against Coleoptera (Leptinotarsa decemlineata, etc.), Diptera (Liriomyza sativae Blanchard, etc.), Isoptera (Termite) pests.

The market for diamide-based and oxadiazine-based insecticides has grown rapidly in recent years, and still has a large potential for future growth. The most widely used of these two classes of insecticides are chlorantraniliprole and indoxacarb. In 2019, chlorantraniliprole took the leading position in the global insecticide market, with global sales of US$1.581 billion; indoxacarb's also reached US$206 million. However, with the widespread use of these two insecticides, a variety of pests have developed varying degrees of resistance to them, which has become the main limiting factor for both insecticides. Resistance monitoring shows that the population of Plutella xylostella in the field has developed a high level of resistance to chlorantraniliprole, and the population resistance in some areas has even reached more than 1000 times; the resistance to indoxacarb is generally more than 10 times, and resistance of some population is more than 100 times. The intensification of insecticide resistance not only seriously affects the life cycle of pesticides, but also leads to an increase in the frequency and dosage of pesticides use, resulting in huge economic losses. Effective control of pest resistance has become an urgent problem in crop protection worldwide.

The resistance mechanisms of pests to insecticides can be basically classified into three categories: metabolic resistance, penetration resistance and target resistance, of which metabolic resistance mediated by detoxification enzymes is more common. Many studies have shown that, glutathione S-transferases (GSTs), as an important detoxification enzyme system in insects, can participate in insects' resistance against commonly used insecticides, such as organochlorines, organophosphates, pyrethroids, neonicotinoids, diamides and abamectin through gene mutation, increased activity, and up-regulated expression. For example, silencing LmGSTs5 and LmGSTu1 in Locusta migratoria significantly increased the sensitivity of its nymphs to malathion and chlorpyrifos. There is also evidence that insect GSTs can directly metabolize a variety of insecticides. For example, the recombinant protein from Helicoverpa armigera HaGST-8 has good metabolic activity to chlorpyrifos, dichlorvos and cypermethrin.

GSTs play an important role in the occurrence and development of pest resistance, and reducing their activity will significantly reduce the resistance of pests. Compounds such as S-Hexyl glutathione (GTX) and diethyl maleate (DEM) have been reported to increase control effects by inhibiting GSTs activity and delaying the metabolism of insecticides by pests. As inhibitors of GSTs, these compounds usually have no insecticidal activity by themselves, but when mixed with insecticides, they can significantly improve the toxicity or efficacy of insecticides, and are important pesticide synergists. The research and development of GSTs-targeted inhibitors can not only improve the control of insecticides, but also delay or reduce insecticide resistance and prolong the life cycle of insecticides, which is of great significance to the management of insecticide resistance.

SUMMARY OF THE INVENTION

The present invention provides a novel inhibitor targeting pest GSTs. The inhibitor has broad inhibitory activities against GSTs in various pests, and can effectively delay the in vivo metabolism of insecticides by GSTs in pests, thereby reducing the metabolic resistance of pests to insecticides. The inhibitor effectively decreases the resistance of resistant strains of Plutella xylostella to ryanodine receptor modulators represented by chlorantraniliprole and voltage-dependent sodium ion channel blockers represented by indoxacarb, making it suitable for use as a synergist for insecticides.

It is an object of the present invention to provide a diphenylpyrazole-based compound of following formula (I), or its pesticidally acceptable salts.

It is another object of the present invention to provide a method for preparing the diphenylpyrazole-based compound of following formula (I), or its pesticidally acceptable salts.

It is another object of the present invention to provide use of the diphenylpyrazole-based compound of following formula (I), or its pesticidally acceptable salts in the preparation of pesticides (for example, preferably synergist for insecticides).

The present invention provides a diphenylpyrazole-based compound of following formula (I), or its pesticidally acceptable salts,

[Image Omitted]

- wherein
- R¹can be:
- (1) an amido group —NH—CH(O) which is unsubstituted or substituted by one or more of the following substituents;
  - (a) —C_1˜6alkyl-R³, —C_3˜8cycloalkyl-R³, —C_2˜6alkenyl-R³, —C_2˜6alkynyl-R³, —NH—R³, —N(R³)₂, —C(O)—R³, —NH—C_1˜6alkyl-R³, —C_1˜6alkyl-NH—R³, —C_1˜6alkyl-N(R³)₂, —C_1˜6alkyl-OR³, —C_3˜8cycloalkyl-OR ³, —OC_1˜6alkyl-R³, —C_1˜6alkyl-OC_1˜6alkyl-R³, —C(O)—NH—R³, —C(O)—N(R³)₂, —NH—C(O)—R³, —C_1˜6alkyl-NH—C(O)—R³, —C_1˜6alkyl-NH—C(O)—OR ³, —NH—C(O)—C_1˜6alkyl-R³, —NH—C(O)—C_1˜6alkyl-OR³, —C(O)—NH—C_1˜6alkyl-R³or —C_1˜6alkyl-SR³;
  - R³is each independently selected from: H, O, S, ═NH, amino, halogen, cyano, —C_1˜6alkyl, —C_3˜8cycloalkyl, —S—C_1˜6alkyl, —S—OH, —SO₂—C_1˜6alkyl, -6˜14 membered aryl, -5˜14 membered heterocyclic group, -5˜14 membered heteroaryl and -adamantyl; R³is unsubstituted or substituted by one or more of the following substituents: O, —OH, halogen, cyano, nitro, —CH(O), amino, —C_1˜6alkyl, —C_2˜6alkenyl, —C_2˜6alkynyl, —OC_1˜6alkyl, nitro, sulfonic acid group, —C_1˜6alkyl-halogen, —C_1˜6alkyl-HS, —C_1˜6alkyl-NH₃⁺, —C_1˜6alkyl-OH, —C_1˜6alkyl-NH—C_1˜6alkyl, or —C_1˜6alkyl-N(C_1˜6alkyl)₂;
  - (b) -6˜14 membered aryl, -5˜14 membered heterocyclic group or -5˜14 membered heteroaryl;
  - the 6˜14 membered aryl, the 5˜14 membered heterocyclic group or the 5˜14 membered heteroaryl is unsubstituted or substituted by one or more of the following substituents: O, —OH, halogen, cyano, nitro, —CH(O), amino, —C_1˜6alkyl, —OC_1˜6alkyl, —C_1˜6alkyl-NH—C_1˜6alkyl, —C_1˜6alkyl-N(C_1˜6alkyl)₂, —C(O)—OC_1˜6alkyl, —NH—C(O)—C_1˜6alkyl, —SO₂—C_1˜6alkyl, —SO₂—NH₂, —SO₂—NH—C_1˜6alkyl, —SO₂—N(C_1˜6alkyl)₂, -6˜14 membered aryl, -5˜14 membered heterocyclic group, or -5˜14 membered heteroaryl;
- (2) N atom and C atom of the amido group —NH—CH(O) are connected to form a ring structure via —C_3˜6alkylene-, —NH—C_2˜6alkylene-, —NH—C(O)—C_1˜6alkylene-, —C_1˜6alkylene-NH—C(O)—, —NH—C_1˜6alkylene-C(O)— or —C_1˜6alkylene-C(O)—; the ring structure is optionally substituted by one or more of the following substituents: —C_1˜6alkyl, —C_3˜8cycloalkyl, —C_3˜8cycloalkyl-C_1˜6alkyl, —OC_1˜6alkyl, -6˜14 membered aryl, -5˜14 membered heterocyclic group, -5˜14 membered heteroaryl, —C_1˜6alkyl-6˜14 membered aryl, —C_1˜6alkyl-5˜14 membered heterocyclic group, —C_1˜6alkyl-5˜14 membered heteroaryl or trifluoroethyl; the 6˜14 membered aryl, the 5˜14 membered heterocyclic group and the 5˜14 membered heteroaryl are optionally substituted by —OC_1˜6alkyl; or, the ring structure is further combined with 5˜8 membered aryl, 5˜8 membered heterocyclic group or 5˜8 membered heteroaryl to fuse into a fused ring.
- the heterocyclic group (including 5˜14 members, 5˜10 members and 5˜8 members) contains 1˜4 heteroatoms selected from N, S and O; the heteroaryl (including 5˜14 members, 5˜10 members and 5˜8 members) contains 1˜4 heteroatoms selected from N, S and O.
- R²can be: —H, -halogen, -amino, —NO₂, —CF, —C_1˜6alkyl, —C_1˜6alkyl-OH, —O—R⁴, —C(O)—R⁴, —C(O)—NH₂, —NH—C(O)—R⁴, —C(O)—O—R⁴or —C(O)—O—N(R⁴)₂; R⁴can be: H or C_1˜6alkyl.

The substituent of the amide group is connected to the N atom of the amino group and/or connected to the C atom of the carbonyl group; and when there are two substituents, the two substituents may be the same or different.

The term “halogen” refers to fluorine, chlorine, bromine or iodine as a substituent. When a halogen atom is used as a substituent, the number of substitutions is one or more, including 1, 2 or 3, etc.

The term “alkyl” refers to a straight-chain or branched-chain alkyl group derived from an alkane by removing one hydrogen atom.

The term “alkenyl” refers to a straight-chain or branched-chain or cyclic alkenyl group containing carbon-carbon double bond.

The term “alkynyl” refers to a straight-chain or branched-chain or cyclic alkynyl group containing carbon-carbon triple bond.

The term “cycloalkyl” is a fully hydrogenated non-aromatic ring composed of mono-, bi- or tri-cyclic rings. Thus, a cycloalkyl can typically be a monocyclic ring containing 3 to 7 ring atoms.

Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Also, “cycloalkyl” includes bridged bicycloalkyl systems.

The term “alkyloxy” refers to a group derived from “alkyl” by connecting through —O— to other moieties.

The term “aryl” refers to a cyclic aromatic group whose ring atoms are carbon atoms, including single-ring aryl and condensed-ring aryl. A single-ring aryl refers to a fully unsaturated aryl, and a condensed-ring aryl refers to a cyclic group formed by two or more ring structures sharing two adjacent carbon atoms with each other, and at least one ring is fully unsaturated aromatic ring.

The term “heterocyclic group” refers to a saturated or unsaturated non-aromatic group consisting of 1 to 3 rings containing 1, 2, 3 or 4 heteroatoms (N, O or S).

The term “heteroaryl” refers to an aromatic ring structure, including monocyclic heteroaryl and condensed ring heteroaryl. wherein at least one of the ring atoms is a heteroatom (N, O or S) and the remaining ring atoms are independently selected from carbon, oxygen, nitrogen and sulfur.

Preferably, R¹can be:

- (1) an amido group —NH—CH(O) which is substituted by one or more of the following substituents,
  - (a) —C_1˜3alkyl-R³, —C_3˜6cycloalkyl-R³, —C_2˜5alkenyl-R³, —C_1˜3alkyl-NH—R³, —C_1˜3alkyl-O-R³, or —O—C_1˜3alkyl-R³; R³are each independently selected from: H, O, S, ═NH, amino, halogen, cyano, —C_1˜3alkyl, —C_3˜6cycloalkyl, -6˜14 membered aryl, -5˜14 membered heterocyclic group, or -5˜14 membered heteroaryl; R³is unsubstituted or substituted by one or more of the following substituents: O, —OH, halogen, cyano, nitro, —CH(O), —S—OH, amino, —C_1˜3alkyl, —C_2˜5alkenyl, —C_2˜5alkynyl, —OC_1˜3alkyl, nitro, sulfonic acid group, —C_1˜3alkyl-halogen or —C_1˜3alkyl-OH;
  - (b) -6˜14 membered aryl, -5˜14 membered heterocyclic group or -5˜14 membered heteroaryl; the 6˜14 membered aryl, the 5˜14 membered heterocyclic group or the 5˜14 membered heteroaryl is unsubstituted or substituted by one or more of the following substituents: O, —OH, halogen, cyano, nitro, —CH(O), amino, —C_1˜3alkyl, —OC_1˜3alkyl, —C_1˜3alkyl-NH—C_1˜3alkyl, —C_1˜3alkyl-N(C_1˜3alkyl)₂, —C(O)—OC_1˜3alkyl, —NH—C(O)—C_1˜3alkyl, —SO₂—C_1˜3alkyl, —S₂—NH₂, —SO₂—NH—C_1˜3alkyl, or —SO₂—N(C_1˜3alkyl)₂;
- (2) N atom and C atom of the amido group —NH—CH(O) are connected to form a ring structure via —C_3˜6alkylene-, —NH—C_2˜4alkylene-, —NH—C(O)—C_1˜3alkylene-, —C_1˜3alkylene-NH—C(O)—, —NH—C_1˜3alkylene-C(O)— or —C_1˜3alkylene-C(O)—; the ring structure is optionally substituted by the following substituents: —C_1˜3alkyl, —C_3˜6cycloalkyl, —C_3˜6cycloalkyl-C_1˜3alkyl, -6˜14 membered aryl, -5˜14 membered heterocyclic group, -5˜14 membered heteroaryl, —C_1˜3alkyl-6˜14 membered aryl, —C_1˜3alkyl-5˜14 membered heterocyclic group, —C_1˜3alkyl-5˜14 membered heteroaryl, or trifluoroethyl; the 6˜14 membered aryl, the 5˜14 membered heterocyclic group and the 5˜14 membered heteroaryl are optionally substituted by —OC_1˜3alkyl;
- the heterocyclic group contains 1˜3 heteroatoms selected from N, S and O; the heteroaryl contains 1˜3 heteroatoms selected from N, S and O.
- R²is: —H or halogen.
- more preferably, R¹can be:
- (1) —NH—C(O)—C_1˜3alkyl-R³; R³is selected from: -6˜10 membered aryl, -5˜10 membered heterocyclic group, -5˜10 membered heteroaryl; R³is unsubstituted or substituted by one or more of the following substituents: O, —OH, halogen, cyano, nitro, —CH(O), —S—OH, amino, —C_1˜3alkyl, —OC_1˜3alkyl, nitro, sulfonic acid group, —C_1˜3alkyl-halogen, —C_1˜3alkyl-HS, —C_1˜3alkyl-NH₃or —C_1˜3alkyl-OH;
- (2) —NH—C(O)-6˜10 membered aryl, —NH—C(O)-5˜10 membered heterocyclic group or —NH—C(O)-5˜10 membered heteroaryl; the 6˜10-membered aryl, the 5˜10-membered heterocyclic group or the 5˜10-membered heteroaryl is unsubstituted or substituted by one or more of the following substituents: O, —OH, halogen, cyano, nitro, —CH(O), amino, —C_1˜3alkyl or —OC_1˜3alkyl;

The heterocyclic group contains 1˜3 heteroatoms selected from N, S and O; the heteroaryl contains 1˜3 heteroatoms selected from N, S and O;

- R²can be: —H.

Specifically, R¹can be:

[Image Omitted]

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Preferably, R¹can be specifically:

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Further preferably, R¹can be specifically:

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Further preferably, R¹can be specifically:

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Further preferably, R¹can be specifically:

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In a specific embodiment, the diphenylpyrazole-based compound o following formula (I) or its pesticidally acceptable salts is the following compound or its pesticidally acceptable salts:

Name
Structure
Number

N′-(1,3-diphenyl-1H-pyrazol-5-yl-carbonyl)-2- oxo-1,2-dihydropyridinyl-4-carbohydrazide

[Image Omitted]

PXG1

N′-[2-(4-methyl-1,3-thiazol-2-yl)acetyl]-1,3- diphenyl-1H-pyrazol-5-yl-carbohydrazide

[Image Omitted]

PXG2

N′-(1,3-diphenyl-1H-pyrazol-5-yl-carbonyl)-2- (phenylamino) acetohydrazide

[Image Omitted]

PXG3

N′-(1,3-diphenyl-1H-pyrazol-5-yl-carbonyl)- 3,4,5-trimethoxybenzohydrazine

[Image Omitted]

PXG4

N′-(1,3-diphenyl-1H-pyrazol-5-yl-carbonyl)-2- phenoxyacetohydrazide

[Image Omitted]

PXG5

N-{2,4-dioxa-1,3-diazaspiro[4.5]dec-3-yl}-1,3- diphenyl-1H-pyrazol-5-yl-carboxamide

[Image Omitted]

PXG6

N′-[3-(4-chlorophenyl)-1-phenyl-1H-pyrazol-5- yl-carbonyl]pyridin-3-yl-carbohydrazide

[Image Omitted]

PXG7

(2S)-N′-(1,3-diphenyl-1H-pyrazol-5-yl- carbonyl)-2,3-dihydro-1,4-benzodioxin-2-yl- carbohydrazide

[Image Omitted]

PXG8

3-[N′-(1,3-diphenyl-1H-pyrazol-5-yl-carbonyl) hydrazinocarbonyl]-N,N-dimethylphenyl-1- sulfonamide

[Image Omitted]

PXG9

N′-(1,3-diphenyl-1H-pyrazol-5-yl-carbonyl)-2- (2-methoxyphenyl) acetohydrazide

[Image Omitted]

PXG10

N′-(1,3-diphenyl-1H-pyrazol-5-yl-carbonyl) benzohydrazine

[Image Omitted]

PXG11

N-(4-methyl-2,5-dioxo-4-phenylimidazolin-1- yl)-1,3-diphenyl-1H-pyrazol-5-yl-carboxamide

[Image Omitted]

PXG12

N-(4-ethyl-4-methyl-2,5-dioxaimidazolin-1-yl)- 1,3-diphenyl-1H-pyrazol-5-yl-carboxamide

[Image Omitted]

PXG13

N-[4-(4-methoxyphenyl)-4-methyl-2,5- dioxaimidazolin-1-yl]-1,3-diphenyl-1H- pyrazol-5-yl-carboxamide

[Image Omitted]

PXG14

N-[4-methyl-2,5-dioxo-4-(2- phenylethyl)imidazolin-1-yl]-1,3-diphenyl-1H- pyrazol-5-yl-carboxamide

[Image Omitted]

PXG15

N-{8-methyl-2,4-dioxo-1,3-diazaspiro[4.5]dec- 3-yl}-1,3-diphenyl-1H-pyrazol-5-yl- carboxamide

[Image Omitted]

PXG16

N-[4-(3-methoxyphenyl)-4-methyl-2,5- dioxaimidazolin-1-yl]-1,3-diphenyl-1H- pyrazol-5-yl-carboxamide

[Image Omitted]

PXG17

N′-(1,3-diphenyl-1H-pyrazol-5-yl-carbonyl)-2- (2-methoxyphenoxy)acetohydrazide

[Image Omitted]

PXG18

2-(4-cyanophenoxy)-N′-(1,3-diphenyl-1H- pyrazol-5-yl-carbonyl)acetohydrazide

[Image Omitted]

PXG19

N′-[3-(4-chlorophenyl)-1-phenyl-1H-pyrazol-5- yl-carbonyl]pyridin-2-yl-carbohydrazide

[Image Omitted]

PXG20

N′-(1-ethyl-2-methyl-1H-1,3-benzodiazol-5-yl- carbonyl)-1,3-diphenyl-1H-pyrazol-5-yl- carbohydrazide

[Image Omitted]

PXG21

N′-(1,3-diphenyl-1H-pyrazol-5-yl-carbonyl)-2- oxo-1,2-dihydroquinoline-4-yl-carbohydrazide

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PXG22

2-(2-cyanophenoxy)-N′-(1,3-diphenyl-1H- pyrazol-5-yl-carbonyl)acetohydrazide

[Image Omitted]

PXG23

N′-(ethoxycarbonyl)-1,3-diphenyl-1H-pyrazol- 5-yl-carbohydrazide

[Image Omitted]

PXG24

N′-(1,3-diphenyl-1H-pyrazol-5-yl-carbonyl)-2- hydroxypyridin-3-yl-carbohydrazide

[Image Omitted]

PXG25

N′-(1,3-diphenyl-1H-pyrazol-5-yl-carbonyl)-2- [(3r)-adamantan-1-yl]acetohydrazide

[Image Omitted]

PXG26

N′-(1,3-diphenyl-1H-pyrazol-5-yl-carbonyl)-6- oxo-1,6-dihydropyridazin-3-yl-carbohydrazide

[Image Omitted]

PXG27

N′-(1,3-diphenyl-1H-pyrazol-5-yl-carbonyl) cyclopentyl carbohydrazide

[Image Omitted]

PXG28

4-[N′-(1,3-diphenyl-1H-pyrazol-5-yl-carbonyl) hydrazinocarbonyl]-N,N-dimethyl phenyl-1- sulfonamide

[Image Omitted]

PXG29

In the most preferably specific embodiment, the diphenylpyrazole-based compound of following formula (I) or its pesticidally acceptable salts is the following compound or its pesticidally acceptable salts:

Num-

Structure
ber

[Image Omitted]

PXG1

PXG2

PXG22

The present invention provides a method for preparing a diphenylpyrazole-based compound of following formula (I), or its pesticidally acceptable salts, which includes steps o:

[Image Omitted]

- hydrolyzing compound A to its carboxylic acid form compound B, and then condensing compound B and compound H₂N—R¹to compound (I) through a condensing agent in a solvent by adding an organic base or an inorganic base, under stirring at room temperature or heating; wherein R¹and R²are defined as above;
- or,

[Image Omitted]

- reacting compound A with hydrazine hydrate to give hydrazide compound C, or hydrolyzing compound A to compound B, then reacting compound B with hydrazine hydrate to give hydrazide compound C, and then condensing compound C and compound COOH—R⁵to compound (I) through a condensing agent in a solvent by adding an organic base or an inorganic base, under stirring at room temperature or heating; wherein R²is defined as above, and R⁵is: (a) —C_1˜6alkyl-R³, —C_3˜8cycloalkyl-R³, —C_2˜6alkenyl-R³, —C_2˜6alkynyl-R³, —NH—R³, —N(R³)₂, —C(O)—R³, —NH—C_1˜6alkyl-R³, —C_1˜6alkyl-NH—R³, —C_1˜6alkyl-N(R³)₂, —C_1˜6alkyl-OR³, —C_3˜8cycloalkyl-O—R³, —O—C_1˜6alkyl-R³, —C_1˜6alkyl-O—C_1˜6alkyl-R³, —C(O)—NH—R³, —C(O)—N(R³)₂, —NH—C(O)—R³, —C_1˜6alkyl-NH—C(O)—R³, —C_1˜6alkyl- NH—C(O)—O—R³, —NH—C(O)—C_1˜6alkyl-R³, —NH—C(O)—C_1˜6alkyl-O—R³, —C(O)—NH—C_1˜6alkyl-R³or —C_1˜6alkyl-S—R³;
- R³is each independently selected from: H, O, S, ═NH, amino, halogen, cyano, —C_1˜6alkyl, —C_3˜8cycloalkyl, —S—C_1˜6alkyl, —S—OH, —SO₂—C_1˜6alkyl, -6˜14 membered aryl, -5˜14 membered heterocyclic group, -5˜14 membered heteroaryl and -adamantyl; R³is unsubstituted or substituted by one or more of the following substituents: O, —OH, halogen, cyano, nitro, —CH(O), amino, —C_1˜6alkyl, —C_2˜6alkenyl, —C_2˜6alkynyl, —O—C_1˜6alkyl, nitro, sulfonic acid group, —C_1˜6alkyl-halogen, —C_1˜6alkyl-HS, —C_1˜6alkyl-NH₃⁺, —C_1˜6alkyl-OH, —C_1˜6alkyl-NH—C_1˜6alkyl or —C_1˜6alkyl-N(C_1˜6alkyl)₂; (b) -6˜14 membered aryl, -5˜14 membered heterocyclic group or -5˜14 membered heteroaryl; said 6˜14 membered aryl, said 5˜14 membered heterocyclic group or said 5˜14 membered heteroaryl is unsubstituted or substituted by one or more of the following substituents: O, —OH, halogen, cyano, nitro, —CH(O), amino, —C_1˜6alkyl, —O—C_1˜6alkyl, —C_1˜6alkyl-NH—C_1˜6alkyl, —C_1˜6alkyl-N(C_1˜6alkyl)₂, —C(O)—O—C_1˜6alkyl, —NH—C(O)—C_1˜6alkyl, —SO₂—C_1˜6alkyl, —SO₂—NH₂, —SO₂—NH—C_1˜6alkyl, —SO₂—N(C_1˜6alkyl)₂, -6˜14 membered aryl, -5˜14 membered heterocyclic group, or -5˜14 membered heteroaryl; said heterocyclic group contains 1˜4 heteroatoms selected from N, S and O; said heteroaryl contains 1 to 4 heteroatoms selected from N, S and O.

The condensing agents include, but are not limited to: active esters, carbodiimides, onium salts, organic phosphorus, and other condensing agents.

The solvents include, but are not limited to: N,N-dimethylformamide, dichloromethane, acetonitrile, and tetrahydrofuran.

The organic bases include, but are not limited to: triethylamine, diisopropylethylamine and the like.

The inorganic base includes but not limited to: sodium carbonate, potassium carbonate, sodium hydroxide, sodium bicarbonate and the like.

The synthesis of Compound A was carried out according to reference (J. Org. Chem. 2010, 75, 3, 984-987).

The present invention provides a composition comprising a diphenylpyrazole-based compound of following formula (I), or its pesticidally acceptable salts, and pesticidally acceptable excipients.

Preferably, the composition contains one or more insecticides.

The present invention also provides use of the diphenylpyrazole-based compound of following formula (I), or its pesticidally acceptable salts in preparation of GSTs inhibitors.

Preferably, the GSTs include: PxGSTδ1, PxGSTε3, PxGSTσ1, PxGSTσ2, PxGSTω4, PxGSTθ1, PxGSTζ1 and PxGSTμ1; preferably, the GSTs include: PxGSTδ1, PxGSTσ1, PxGSTσ2 and PxGSTε3; more preferably, the GSTs include: PxGSTδ1 and PxGSTε3.

The present invention also provides use of the diphenylpyrazole-based compound of following formula (I), or its pesticidally acceptable salts in the preparation of pesticides, preferably insecticide synergists.

The insecticide synergist delays or reduces pest resistance to insecticides.

The insecticides include: ryanodine receptor modulator insecticides (for example, diamide insecticides); voltage-dependent sodium ion channel blocker insecticides (for example: oxadiazine insecticides).

Specifically, the insecticides include: chlorantraniliprole and indoxacarb.

The pests include: field crop pests and economic crop pests.

Specifically, the pest include: Plutella xylostella, Mythimna separata, Pyrausta nubilalis, Chilo suppressalis, Nilaparvata lugens, Spodoptera frugiperda, Helicoverpa armigera, Carposina sasakii, and Spodoptera litura.

The present invention provides a class of insect GSTs inhibitors with novel structures. It is found that the compounds with such structures have broad inhibitory activity on GSTs in insects, and significantly improve the sensitivity of resistant pest strains to various insecticides and the effectiveness of insecticides. The compounds play an important role in managing the insecticide resistance of pests.

DESCRIPTION OF THE DRAWINGS

FIG. 1 is the mass spectrogram of the compound PXG1 prepared in the present invention.

FIG. 2 is the mass spectrogram of the compound PXG2 prepared in the present invention.

FIG. 3 is the mass spectrogram of the compound PXG3 prepared in the present invention.

FIG. 4 is the ¹H-NMR spectrogram of the compound PXG3 prepared in the present invention dissolved in deuterated DMSO.

FIG. 5 is the mass spectrogram of the compound PXG4 prepared in the present invention.

FIG. 6 is the ¹H-NMR spectrogram of the compound PXG4 prepared in the present invention dissolved in deuterated DMSO.

FIG. 7 is the mass spectrogram of the compound PXG5 prepared in the present invention.

FIG. 8 is the mass spectrogram of the compound PXG6 prepared in the present invention.

FIG. 9 is the ¹H-NMR spectrogram of the compound PXG6 prepared in the present invention dissolved in deuterated DMSO.

FIG. 10 is the mass spectrogram of the compound PXG7 prepared in the present invention.

FIG. 11 is the mass spectrogram of the compound PXG8 prepared in the present invention.

FIG. 12 is the mass spectrogram of the compound PXG9 prepared in the present invention.

FIG. 13 is the mass spectrogram of the compound PXG10 prepared in the present invention.

FIG. 14 is the ¹H-NMR spectrogram of the compound PXG10 prepared in the present invention dissolved in deuterated DMSO.

FIG. 15 is the mass spectrogram of the compound PXG11 prepared in the present invention.

FIG. 16 is the ¹H-NMR spectrogram of the compound PXG11 prepared in the present invention dissolved in deuterated DMSO.

FIG. 17 is the mass spectrogram of the compound PXG12 prepared in the present invention.

FIG. 18 is the mass spectrogram of the compound PXG13 prepared in the present invention.

FIG. 19 is the mass spectrogram of the compound PXG14 prepared in the present invention.

FIG. 20 is the mass spectrogram of the compound PXG15 prepared in the present invention.

FIG. 21 is the mass spectrogram of the compound PXG16 prepared in the present invention.

FIG. 22 is the ¹H-NMR spectrogram of the compound PXG16 prepared in the present invention dissolved in deuterated DMSO.

FIG. 23 is the mass spectrogram of the compound PXG17 prepared in the present invention.

FIG. 24 is the mass spectrogram of the compound PXG18 prepared in the present invention.

FIG. 25 is the ¹H-NMR spectrogram of the compound PXG18 prepared in the present invention dissolved in deuterated DMSO.

FIG. 26 is the mass spectrogram of the compound PXG19 prepared in the present invention.

FIG. 27 is the ¹H-NMR spectrogram of the compound PXG19 prepared in the present invention dissolved in deuterated DMSO.

FIG. 28 is the mass spectrogram of the compound PXG20 prepared in the present invention.

FIG. 29 is the mass spectrogram of the compound PXG21 prepared in the present invention.

FIG. 30 is the mass spectrogram of the compound PXG22 prepared in the present invention.

FIG. 31 is the ¹H-NMR spectrogram of the compound PXG22 prepared in the present invention dissolved in deuterated DMSO.

FIG. 32 is the mass spectrogram of the compound PXG23 prepared in the present invention.

FIG. 33 is the ¹H-NMR spectrogram of the compound PXG23 prepared in the present invention dissolved in deuterated DMSO.

FIG. 34 is the mass spectrogram of the compound PXG24 prepared in the present invention.

FIG. 35 is the ¹H-NMR spectrogram of the compound PXG24 prepared in the present invention dissolved in deuterated DMSO.

FIG. 36 is the mass spectrogram of the compound PXG25 prepared in the present invention.

FIG. 37 is the mass spectrogram of the compound PXG26 prepared in the present invention.

FIG. 38 is the ¹H-NMR spectrogram of the compound PXG26 prepared in the present invention dissolved in deuterated DMSO.

FIG. 39 is the mass spectrogram of the compound PXG27 prepared in the present invention.

FIG. 40 is the ¹H-NMR spectrogram of the compound PXG27 prepared in the present invention dissolved in deuterated DMSO.

FIG. 41 is the mass spectrogram of the compound PXG28 prepared in the present invention.

FIG. 42 is the mass spectrogram of the compound PXG29 prepared in the present invention.

FIGS. 43A-43D are the inhibition curve of some compounds of the present invention against total enzymes of GSTs of Plutella xylostella.

FIGS. 44A-44B are the inhibition curve of some compounds of the present invention against total enzymes of GSTs of Carposina sasakii.

FIGS. 45A-45C are the inhibition curve of some compounds of the present invention against total enzymes of GSTs of Spodoptera litura.

FIGS. 46A-46E are the inhibition curve of some compounds of the present invention against total enzymes of GSTs of Mythimna separata.

FIGS. 47A-47D are the inhibition curve of some compounds of the present invention against GSTs total enzymes of Pyrausta nubilalis.

FIGS. 48A-48G are the inhibition curve of some compounds of the present invention against GSTs total enzymes of Chilo suppressalis.

FIGS. 49A-49E are the inhibition curve of some compounds of the present invention against GSTs total enzymes of Nilaparvata lugens.

FIGS. 50A-50D are the the inhibition curve of some compounds of the present invention against GSTs total enzymes of Spodoptera frugiperda.

FIGS. 51A-51D are the inhibition curve of some compounds of the present invention against GSTs total enzymes of Helicoverpa armigera.

FIGS. 52A-52D are the enzyme kinetics curve of Pxlutella xylostella PxGSTs recombinant protein.

FIGS. 53A-53N are the inhibition curve of some compounds of the present invention against PxGSTδ1.

FIGS. 54A-54B are the inhibition curve of some compounds of the present invention against PxGSTε3.

FIGS. 55A-55I are the inhibition curve of some compounds of the present invention against PxGSTσ1.

FIGS. 56A-56C are the inhibition curve of some compounds of the present invention against PxGSTσ2.

DETAILED EMBODIMENTS

The present invention will be described below through examples, but the present invention is not limited thereto. The experimental methods shown in the following examples are conventional methods unless otherwise specified. The reagents and materials shown are all commercially available.

Preparation Example 1 Preparation of Compound PXG1

[Image Omitted]

Method 1:

(1) Preparation of hydrazide compound C1: 10 g of compound A1 (A1 was prepared by reference to J. Org. Chem. 2010, 75, 3, 984-987; HNMR (400 MHz, CDCl₃): δ 3.83 (s, 3H), 7.33-7.36 (m, 2H), 7.41-7.50 (m, 7H), 7.87 (m, 2H). the same below), 100 ml of absolute ethanol, 20 ml of hydrazine hydrate was first added into 100 ml three-necked flask, heated to reflux overnight, cooled and filtered by suction to obtain 9.5 g of compound C1, yield 95%.

(2) Compound F1 (carboxylic acid) (2.5 g, 1 eq), DMF (10 ml), EDCI (4.14 g, 1.2 eq), potassium carbonate (6.21 g, 2.5 eq) were added into 100 ml three-necked flask, stirred for 10 minutes and then added with compound C1 (hydrazide) (5.5 g, 1.1 eq), stirred overnight at room temperature, the reaction was monitored by TLC until completion. The resultant was diluted with water, the formed precipitate was filtered off and dried, the crude compound was purified by reverse phase HPLC without additional treatment with the eluent methanol-water, the selected fractions were combined and concentrated to obtain the target product PXG1.

Or, method 2:

(1) Preparation of compound B1: 10 g of compound A1, 100 ml of absolute ethanol, 28.7 ml (2 eq) of 10% NaOH was first put into 100 ml three-necked flask, heated to reflux for 5 hours, the reaction was completely cooled, and was adjusted to pH=2-3 with 10% hydrochloric acid, filtered by suction to obtain 8 g of compound B1, yield 85%.

(2) Compound B1 (carboxylic acid) (6.5 g, 1 eq), DMF (10 ml), EDCI (5.75 g, 1.2 eq), potassium carbonate (8.63 g, 2.5 eq) were added into 100 ml three-necked flask, stirred for 10 minutes and then added with compound G1 (hydrazide) (4.21 g, 1.1 eq), stirred overnight at room temperature, the reaction was monitored by TLC until completion. The resultant was diluted with water, the formed precipitate was filtered off and dried, the crude compound was purified by reverse phase HPLC without additional treatment with the eluent methanol-water, the selected fractions were combined and concentrated to obtain the target product PXG1.

MS(ES-API) cacld. for C₂₂H₁₇N₅O₃found 400.2 [M+1]⁺. (FIG. 1)

Preparation Example 2 Preparation of Compound PXG2

[Image Omitted]

Method 1: Compound PXG2 was prepared using the same method and steps as in Preparation Example 1, except that compound F2 (carboxylic acid) (2.83 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or,

Method 2: Compound PXG2 was prepared using the same method and steps as in Preparation Example 1, except that compound G2 (hydrazide) (4.70 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₂H₁₉N₅O₂S found 418.2 [M+1]⁺. (FIG. 2)

Preparation Example 3 Preparation of Compound PXG3

[Image Omitted]

Method 1: Compound PXG3 was prepared using the same method and steps as in Preparation Example 1, except that compound F3 (carboxylic acid) (2.72 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or,

Method 2: Compound PXG3 was prepared using the same method and steps as in Preparation Example 1, except that compound G3 (hydrazide) (4.54 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₄H₂₁N₅O₂found 412.2 [M+1]⁺. (FIG. 3); HNMR (400 MHz, DMSO): δ 3.6 (s, 2H), 2.45 (m, 5H), 5.65 (s, 1H), 6.6 (m, 3H), 7.05-7.2 (m, 3H), 7.4-7.5 (m, 12H), 10.0 (s, 2H). (FIG. 4)

Preparation Example 4 Preparation of Compound PXG4

[Image Omitted]

Method 1: Compound PXG4 was prepared using the same method and steps as in Preparation Example 1, except that compound F4 (carboxylic acid) (3.81 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or,

Method 2: Compound PXG4 was prepared using the same method and steps as in Preparation Example 1, except that compound G4 (hydrazide) (6.22 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₆H₂₄N₄O₅found 473.2 [M+1]⁺. (FIG. 5); HNMR (400 MHz, DMSO): δ 3.75 (s, 3H), 3.9 (s, 6H), 7.05 (s, 1H), 7.2-7.5 (m, 12H), 8.05 (s, 1H), 10.0 (s, 1H), 10.4 (s, 1H). (FIG. 6)

Preparation Example 5 Preparation of Compound PXG5

[Image Omitted]

Method 1: Compound PXG5 was prepared using the same method and steps as in Preparation Example 1, except that compound F5 (carboxylic acid) (2.74 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or,

Method 2: Compound PXG5 was prepared using the same method and steps as in Preparation Example 1, except that compound G5 (hydrazide) (4.57 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₄H₂₀N₄O₃found 413.2 [M+1]⁺. (FIG. 7)

Preparation Example 6 Preparation of Compound PXG6

[Image Omitted]

Method 1: Compound PXG6 was prepared using the same method and steps as in Preparation Example 1, except that compound G6 (hydrazide) (5.04 g, 1 eq) was added into the 100 ml three-neck flask in step (2).

MS(ES-API) cacld. for C₂₄H₂₃N₅O₃found 430.2 [M+1]+. (FIG. 8); HNMR (400 MHz, DMSO): δ 1.4 (m, 1H), 1.5-1.9 (m, 9H), 7.05 (s, 1H), 7.2-7.4 (m, 10H), 8.8 (s, 1H), 10.6 (s, 1H). (FIG. 9)

Preparation Example 7 Preparation of Compound PXG7

[Image Omitted]

Method 1:

(1) Preparation of hydrazide compound C2: 10 g of compound A2 (A2 was prepared by reference to J Org. Chem. 2010, 75, 3, 984-987), 100 ml of absolute ethanol, 20 ml of hydrazine hydrate was first added into 100 ml three-necked flask, heated to reflux overnight, cooled and filtered by suction to obtain 7.5 g of compound C2, yield 75%.

(2) Compound F7 (carboxylic acid) (2.10 g, 1 eq), DMF (10 ml), EDCI (4.14 g, 1.2 eq), potassium carbonate (6.21 g, 2.5 eq) were added into 100 ml three-necked flask, stirred for 10 minutes and then added with compound C2 (hydrazide) (6.0 g, 1.1 eq), stirred overnight at room temperature, the reaction was monitored by TLC until completion. The resultant was diluted with water, the formed precipitate was filtered off and dried, the crude compound was purified by reverse phase HPLC without additional treatment with the eluent methanol-water, the selected fractions were combined and concentrated to obtain the target product PXG7.

Method 2:

(1) Preparation of compound B2: 10 g of compound A2, 100 ml of absolute ethanol, 28.7 ml (2 eq) of 10% NaOH was first added into 100 ml three-necked flask, heated to reflux for 5 hours, the reaction was completely cooled, and was adjusted to pH=2-3 with 10% hydrochloric acid, filtered by suction to obtain 6.8 g of compound B2, yield 70%.

(2) Compound B2 (carboxylic acid) (7.0 g, 1 eq), DMF 10 ml, EDCI (5.75 g, 1.2 eq), potassium carbonate (8.63 g, 2.5 eq) were added into 100 ml three-necked flask, stirred for 10 minutes and then added with compound G7 (hydrazide) (3.48 g, 1.1 eq), stirred overnight at room temperature, the reaction was monitored by TLC until completion. The resultant was diluted with water, the formed precipitate was filtered off and dried, the crude compound was purified by reverse phase HPLC without additional treatment with the eluent methanol-water, the selected fractions were combined and concentrated to obtain the target product PXG7.

MS(ES-API) cacld. for C₂₂H₁₆ClN₅O₂found 418.0[M+1]⁺. (FIG. 10)

Preparation Example 8 Preparation of Compound PXG8

[Image Omitted]

Method 1: Compound PXG8 was prepared using the same method and steps as in Preparation Example 1, except that compound F8 (carboxylic acid) (3.24 g, 1 eq) was added into the 100 ml three-neck flask in step (2).

Method 2: Compound PXG8 was prepared using the same method and steps as in Preparation Example 1, except that compound G8 (hydrazide) (5.34 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₅H₂₀N₄O₄found 441.0[M+1]⁺. (FIG. 11)

Preparation Example 9 Preparation of Compound PXG9

[Image Omitted]

Method 1: Compound PXG9 was prepared using the same method and steps as in Preparation Example 1, except that compound F9 (carboxylic acid) (4.12 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or

Method 2: Compound PXG9 was prepared using the same method and steps as in Preparation Example 1, except that compound G9 (hydrazide) (6.68 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₅H₂₃N₅O₄S found 490.2[M+1]⁺. (FIG. 12)

Preparation Example 10 Preparation of Compound PXG10

[Image Omitted]

Method 1: Compound PXG10 was prepared using the same method and steps as in Preparation Example 1, except that compound F10 (carboxylic acid) (3.00 g, 1 eq) was added into the 100 ml three-neck flask in step (2).

Method 2: Compound PXG10 was prepared using the same method and steps as in Preparation Example 1, except that compound G10 (hydrazide) (4.95 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₅H₂₂N₄O₃found 427.2[M+1]⁺. (FIG. 13); HNMR (400 MHz, DMSO): δ 3.5 (s, 2H), 3.8 (s, 3H), 6.8 (m, 2H), 7.05 (s, 1H), 7.2-7.6 (m, 12H), 10.0 (dd, 2H). (FIG. 14)

Preparation Example 11 Preparation of Compound PXG11

[Image Omitted]

Method 1: Compound PXG11 was prepared using the same method and steps as in Preparation Example 1, except that compound F11 (carboxylic acid) (2.20 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or

Method 2: Compound PXG11 was prepared using the same method and steps as in Preparation Example 1, except that compound G11 (hydrazide) (3.74 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₃H₁₈N₄O₂found 383.2[M+1]⁺. (FIG. 15); HNMR (400 MHz, DMSO): δ 7.05 (s, 1H), 7.2-7.6 (m, 13H), 7.95 (s, 2H), 10.05 (s, 1H), 10.45 (s, 1H). (FIG. 16)

Preparation Example 12 Preparation of Compound PXG12

[Image Omitted]

Method 1: Compound PXG12 was prepared using the same method and steps as in Preparation Example 1, except that compound G12 (hydrazide) (5.64 g, 1 eq) was added into the 100 ml three-neck flask in step (2).

MS(ES-API) cacld. for C₂₆H₂₁N₅O₃found 452.2[M+1]⁺. (FIG. 17)

Preparation Example 13 Preparation of Compound PXG13

[Image Omitted]

Method 1: Compound PXG13 was prepared using the same method and steps as in Preparation Example 1, except that compound G13 (hydrazide) (4.32 g, 1 eq) was added into the 100 ml three-neck flask in step (2).

MS(ES-API) cacld. for C₂₂H₂₁N₅O₃found 404.2[M+1]⁺. (FIG. 18)

Preparation Example 14 Preparation of Compound PXG14

[Image Omitted]

Method 1: Compound PXG14 was prepared using the same method and steps as in Preparation Example 1, except that compound G14 (hydrazide) (6.47 g, 1 eq) was added into the 100 ml three-neck flask in step (2).

MS(ES-API) cacld. for C₂₇H₂₃N₅O₄found 482.2[M+1]⁺. (FIG. 19)

Preparation Example 15 Preparation of Compound PXG15

[Image Omitted]

Method 1: Compound PXG15 was prepared using the same method and steps as in Preparation Example 1, except that compound G15 (hydrazide) (6.41 g, 1 eq) was added into the 100 ml three-neck flask in step (2).

MS(ES-API) cacld. for C₂₈H₂₅N₅O₃found 480.2[M+1]+. (FIG. 20)

Preparation Example 16 Preparation of Compound PXG16

[Image Omitted]

Method 1: Compound PXG16 was prepared using the same method and steps as in Preparation Example 1, except that compound G16 (hydrazide) (5.42 g, 1 eq) was added into the 100 ml three-neck flask in step (2).

MS(ES-API) cacld. for C₂₅H₂₅N₅O₃found 444.2[M+1]⁺. (FIG. 21); HNMR (400 MHz, DMSO): δ 1.0 (s, 3H), 1.1-1.8 (m, 8H), 7.05 (s, 1H), 7.2-7.4 (m, 10H), 9.05 (s, 1H), 10.5 (s, 1H). (FIG. 22)

Preparation Example 17 Preparation of Compound PXG17

[Image Omitted]

Method 1: Compound PXG17 was prepared using the same method and steps as in Preparation Example 1, except that compound G17 (hydrazide) (6.47 g, 1 eq) was added into the 100 ml three-neck flask in step (2).

MS(ES-API) cacld. for C₂₇H₂₃N₅O₄found 482.2[M+1]⁺. (FIG. 23)

Preparation Example 18 Preparation of Compound PXG18

[Image Omitted]

Method 1: Compound PXG18 was prepared using the same method and steps as in Preparation Example 1, except that compound F18 (carboxylic acid) (3.28 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or

Method 2: Compound PXG18 was prepared using the same method and steps as in Preparation Example 1, except that compound G18 (hydrazide) (5.39 g, 1.1 eq) added put after stirring for 10 minutes in step (21.

MS(ES-API) cacld. for C₂₅H₂₂N₄O₄found 443.2[M+1]⁺. (FIG. 24); HNMR (400 MHz, DMSO): δ 3.85 (s, 3H), 4.45 (s, 2H), 6.8-7.25 (m, 5H), 7.2-7.5 (m, 10H), 8.05 (s, 1H), 10.25 (s, 1H). (FIG. 25)

Preparation Example 19 Preparation of Compound PXG19

[Image Omitted]

Method 1: Compound PXG19 was prepared using the same method and steps as in Preparation Example 1, except that compound F19 (carboxylic acid) (3.19 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or

Method 2: Compound PXG19 was prepared using the same method and steps as in Preparation Example 1, except that compound G19 (hydrazide) (5.25 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₅H₁₉N₅O₃found 438.2[M+1]+. (FIG. 26); HNMR (400 MHz, DMSO): δ 4.75 (s, 2H), 7.05-7.5 (m, 15H), 7.8 (dd, 2H), 8.05 (s, 1H), 10.5 (s, 1H). (FIG. 27)

Preparation Example 20 Preparation of Compound PXG20

[Image Omitted]

Method 1: Compound PXG20 was prepared using the same method and steps as in Preparation Example 7, except that compound F20 (carboxylic acid) (2.09 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or

Method 2: Compound PXG20 was prepared using the same method and steps as in Preparation Example 7, except that compound G20 (hydrazide) (3.47 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₂H₁₆ClN₅O₂found 418.2[M+1]⁺. (FIG. 28)

Preparation Example 21 Preparation of Compound PXG21

[Image Omitted]

Method 1: Compound PXG21 was prepared using the same method and steps as in Preparation Example 1, except that compound F21 (carboxylic acid) (3.67 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or

Method 2: Compound PXG21 was prepared using the same method and steps as in Preparation Example 1, except that compound G21 (hydrazide) (6.00 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₂H₁₆ClN₅O₂found 465.2[M+1]⁺. (FIG. 29)

Preparation Example 22 Preparation of Compound PXG22

[Image Omitted]

Method 1: Compound PXG22 was prepared using the same method and steps as in Preparation Example 1, except that compound F22 (carboxylic acid) (3.40 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or

Method 2: Compound PXG22 was prepared using the same method and steps as in Preparation Example 1, except that compound G22 (hydrazide) (5.58 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₆H₁₉N₅O₃found 450.2[M+1]⁺. (FIG. 30); HNMR (400 MHz, DMSO): δ 1.05 (s, 1H), 2.45 (m, 5H), 3.2 (s, 5H), 6.6 (s, 1H), 7.05-7.5 (m, 15H), 8.05 (s, 1H), 10.0 (s, 1H), 10.4 (s, 1H), 12.05 (s, 1H). (FIG. 31)

Preparation Example 23 Preparation of Compound PXG23

[Image Omitted]

Method 1: Compound PXG23 was prepared using the same method and steps as in Preparation Example 1, except that compound F23 (carboxylic acid) (3.19 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or

Method 2: Compound PXG23 was prepared using the same method and steps as in Preparation Example 1, except that compound G23 (hydrazide) (5.25 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₅H₁₉N₅O₃found 438.2[M+1]⁺. (FIG. 32); HNMR (400 MHz, DMSO): δ 4.8 (s, 2H), 7.05-7.8 (m, 17H), 10.3 (s, 1H). (FIG. 33)

Preparation Example 24 Preparation of Compound PXG24

[Image Omitted]

Method 1: Compound PXG24 was prepared using the same method and steps as in Preparation Example 1, except that compound F24 (carboxylic acid) (1.62 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or

Method 2: Compound PXG24 was prepared using the same method and steps as in Preparation Example 1, except that compound G24 (hydrazide) (2.86 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₁₉H₁₈N₄O₃found 351.2[M+1]⁺. (FIG. 34); HNMR (400 MHz, DMSO): δ 1.25 (s, 3H), 4.05 (s, 2H), 7.05 (s, 1H), 7.2-7.4 (in, 10H). (FIG. 35)

Preparation Example 25 Preparation of Compound PXG25

[Image Omitted]

Method 1: Compound PXG25 was prepared using the same method and steps as in Preparation Example 1, except that compound F25 (carboxylic acid) (2.50 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or

Method 2: Compound PXG25 was prepared using the same method and steps as in Preparation Example 1, except that compound G25 (hydrazide) (4.21 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₂H₁₇N₅O₃found 400.2[M+1]⁺. (FIG. 36)

Preparation Example 26 Preparation of Compound PXG26

[Image Omitted]

Method 1: Compound PXG26 was prepared using the same method and steps as in Preparation Example 1, except that compound F26 (carboxylic acid) (3.49 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or

Method 2: Compound PXG26 was prepared using the same method and steps as in Preparation Example 1, except that compound G26 (hydrazide) (5.72 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₈H₃₀N₄O₂found 455.2[M+1]⁺. (FIG. 37); HNMR (400 MHz, DMSO): δ 1.6-2.0 (m, 19H), 7.1 (s, 1H), 7.2-7.4 (m, 10H), 9.6 (d, 2H). (FIG. 38)

Preparation Example 27 Preparation of Compound PXG27

[Image Omitted]

Method 1: Compound PXG27 was prepared using the same method and steps as in Preparation Example 1, except that compound F27 (carboxylic acid) (2.52 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or

Method 2: Compound PXG27 was prepared using the same method and steps as in Preparation Example 1, except that compound G27 (hydrazide) (4.24 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₁H₁₆N₆O₃found 455.2[M+1]+. (FIG. 39); HNMR (400 MHz, DMSO): δ 6.8-7.3 (m, 13H), 7.8 (m, 1H), 10.2 (m, 2H), 13.5 (s, 1H). (FIG. 40)

Preparation Example 28 Preparation of Compound PXG28

[Image Omitted]

Method 1: Compound PXG28 was prepared using the same method and steps as in Preparation Example 1, except that compound F28 (carboxylic acid) (2.05 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or

Method 2: Compound PXG28 was prepared using the same method and steps as in Preparation Example 1, except that compound G28 (hydrazide) (3.52 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₂H₂₂N₄O₂found 375.2 [M+1]⁺. (FIG. 41)

Preparation Example 29 Preparation of Compound PXG29

[Image Omitted]

Method 1: Compound PXG29 was prepared using the same method and steps as in Preparation Example 1, except that compound F29 (carboxylic acid) (4.12 g, 1 eq) was added into the 100 ml three-neck flask in step (2). Or

Method 2: Compound PXG29 was prepared using the same method and steps as in Preparation Example 1, except that compound G29 (hydrazide) (6.68 g, 1.1 eq) was added after stirring for 10 minutes in step (2).

MS(ES-API) cacld. for C₂₅H₂₃N₅O₄S found 490.2[M+1]⁺. (FIG. 42)

Experimental Example 1: Inhibitory Activity Assay of the Compounds PXG1-PXG29 (29 Compounds) of the Present Invention Against the Total GSTs Enzyme of 6 Kinds of Field Crop Pests and 3 Kinds of Economic Crop Pests

Experiment: About 50 mg of larvae/nymphs of nine major pests such as Plutella xylostella, Mythimna separata, Pyrausta nubilalis, Chilo suppressalis, Nilaparvata lugens, Spodoptera frugiperda, Helicoverpa armigera, Carposina sasakii, Spodoptera litura were taken, grounded in a mortar pre-cooled at −80° C., and then 1 mL of 100 mM sodium phosphate buffer (1 mM EDTA, pH 7.2) was added and fully homogenized. The homogenate was transferred to a 1.5 mL centrifuge tube at 4° C., and centrifuged at 14,000 g for 30 minutes, and the supernatant was transferred to a new centrifuge tube as the GSTs total enzyme. The protein concentrations of the above total enzymes were determined using the BCA method.

100 μL of PBS buffer (100 mM, pH 7.2) containing 1 mM GSH was taken and placed in a 96-well plate, 5 μg of insect total GSTs enzymes and an appropriate concentration of compounds PXG1-PXG29 (29 compounds) of the present invention were added to each well, after uniformly mixing, and allowed to stand at 30° C. for 10 min. Then 100 μL of PBS buffer (100 mM, pH 7.2) containing 1 mM CDNB was added to each well. Immediately after mixing, the mixture was placed in a microplate reader to measure the absorbance at a wavelength of 340 nm (A 340) once every 1 min, the measurement was carried out continuously for 5 min, the light absorption values of each measuring point were recorded and the inhibition rate in vitro of the compounds with different concentrations against insects total GSTs enzymes were calculated. Each group of assays was repeated three times, and total GSTs enzymes inactivated at 100° C. for 5 min were used as negative controls, and GTX was used as positive controls. GraphPad Prism 5 software was used for analysis and calculation of IC₅₀. Relative inhibition was calculated using the following formula:

Relative
⁢

inhibition
⁢

(
%
)

=

Δ
⁢

A

340
⁢
c

-

Δ
⁢

A

340
⁢
t

Δ
⁢

OD

340
⁢
c

×
100

- wherein, A₃₄₀is the change of A₃₄₀within 5 minutes for the control group, and A_340tis the change of A₃₄₀within 5 minutes for the experimental group.

Results: The experimental results are shown in Table 1, Table 2, and FIGS. 43 to 51. The compounds PXG1-PXG29 (29 compounds) of the present invention have a certain inhibitory activity on the total GSTs enzymes of field crop pests and economic crop pests. Among them, IC₅₀of the compound PXG1 of the present invention on the total GSTs enzymes of field crop pests including Mythimna separata, Pyrausta nubilalis, Chilo suppressalis, Nilaparvata lugens, Spodoptera frugiperda and Helicoverpa armigera, and economic crop pests such as Plutella xylostella, Carposina sasakii and Spodoptera litura is 27.57 μM, 7.2 μM, 12.11 μM, 8.24 μM, 3.62 μM, 62.38 μM, 52.78 μM, 52.63 μM, 17.67 μM, respectively; IC₅₀of the compound PXG2 of the present invention on the total GSTs enzymes of field crop pests such as Mythimna separata, Pyrausta nubilalis, Chilo suppressalis, Nilaparvata lugens, Spodoptera frugiperda and Helicoverpa armigera, and economic crop pests such as Plutella xylostella is 18.09 μM, 15.69 μM, 42.01 μM, 47.88 μM, 2.18 μM, 40.16 μM and 52.13 μM, respectively. IC₅₀of the compound PXG2 of the present invention on the total GSTs enzyme of economic crop pests such as Carposina sasakii and Spodoptera litura is greater than 100 μM, and the inhibitory rates at 100 μM are respectively 35.12% and 57.13%; IC₅₀of the compound PXG22 of the present invention on the total GSTs enzyme of field crop pests such as Mythimna separata, Chilo suppressalis, Nilaparvata lugens, Spodoptera frugiperda and Helicoverpa armigera, and economic crop pests such as Plutella xylostella and Spodoptera litura is 52.98 μM, 19.01 μM, 59.29 μM, 1.72 μM, 77.05 μM, 32.8 μM, 43.1 μM, respectively. IC₅₀of the compound PXG22 of the present invention on the total GSTs enzyme of field crop pests Pyrausta nubilalis, and economic crop pests such as Carposina sasakii are greater than 100 μM, and the inhibitory rates at 100 μM are respectively 42.38% and 42.76%; IC₅₀of the positive control GSTs inhibitor GTX (S-Hexylglutathione) on the total GSTs enzymes of field crop pests such as Mythimna separata, Pyrausta nubilalis, Chilo suppressalis, Nilaparvata lugens, Spodoptera frugiperda and Helicoverpa armigera, and economic crop pests such as Plutella xylostella, Carposina sasakii and Spodoptera litura is 12.9 μM, 16.33 μM, 23.46 μM, 6.65 μM, 2.31 μM, 14.41 μM, 14.05 μM, 6.87 μM, 15.19 μM, respectively.

TABLE 1

The inhibitory effect of compounds PXG1-PXG29 of the present invention on the total

GSTs enzymes of 3 kinds of economic crop pests

Plutella xylostella GSTs

Spodoptera litura GSTs

100 μM

Carposina sasakii GSTs
100 μM

Inhibition
IC₅₀
100 μM
IC₅₀
Inhibition
IC₅₀

Index
rate %
(μM)
Inhibition rate %
(μM)
rate %
(μM)

PXG1
70.90
27.57
71.91
52.63
80.06
17.67

PXG2
69.27
18.09
35.12
>100
57.13
>100

PXG3
25.47
>100
23.07
>100
17.09
>100

PXG4
−10.64
>100
2.34
>100
−17.52
>100

PXG5
11.58
>100
23.22
>100
37.80
>100

PXG 6
21.15
>100
18.21
>100
6.90
>100

PXG 7
15.46
>100
48.15
>100
57.87
>100

PXG 8
35.73
>100
19.83
>100
44.25
>100

PXG 9
3.94
>100
12.55
>100
12.58
>100

PXG 10
−17.71
>100
32.54
>100
3.87
>100

PXG 11
22.90
>100
76.09
>100
24.52
>100

PXG 12
15.08
>100
17.94
>100
4.60
>100

PXG 13
26.60
>100
25.90
>100
9.91
>100

PXG 14
10.33
>100
20.64
>100
2.49
>100

PXG 15
16.83
>100
17.01
>100
−5.22
>100

PXG 16
8.64
>100
6.98
>100
−0.15
>100

PXG 17
−1.63
>100
12.40
>100
−4.14
>100

PXG 18
32.94
>100
13.69
>100
7.95
>100

PXG 19
23.03
>100
19.42
>100
31.70
>100

PXG 20
−1.50
>100
10.99
>100
7.09
>100

PXG 21
16.96
>100
35.52
>100
5.00
>100

PXG 22
60.50
52.98
42.77
>100
56.09
43.1

PXG 23
26.35
>100
19.76
>100
12.89
>100

PXG 24
29.72
>100
22.33
>100
28.38
>100

PXG 25
31.66
>100
48.54
>100
22.37
>100

PXG 26
25.09
>100
18.11
>100
15.89
>100

PXG 27
42.68
>100
43.24
>100
45.08
>100

PXG 28
22.40
>100
49.51
>100
46.09
>100

PXG 29
9.32
>100
33.57
>100
4.60
>100

GTX
73.72
12.9
95.39
6.87
85.61
15.19

TABLE 2

The inhibitory effect of compounds PXG1-PXG29 of the present invention on the total

GSTs enzymes of 9 kinds of field crop pests

Mythimna
Pyrausta
Chilo
Nilaparvata
Spodoptera
Helicoverpa

separata
nubilalis
suppressalis
lugens
frugiperda
armigera

GSTs
GSTs
GSTs
GSTs
GSTs
GSTs

100 μM

100 μM

100 μM

100 μM

100 μM

100 μM

Inhibition
IC₅₀
Inhibition
IC₅₀
Inhibition
IC₅₀
Inhibition
IC₅₀
Inhibition
IC₅₀
Inhibition
IC₅₀

Index
rate %
(μM)
rate %
(μM)
rate %
(μM)
rate %
(μM)
rate %
(μM)
rate %
(μM)

PXG1
95.19
7.20
87.80
12.11
92.32
8.24
100.60
3.62
58.40
62.38
77.22
52.78

PXG2
90.46
15.69
64.92
42.01
52.60
47.88
91.51
2.18
64.72
40.16
57.53
52.13

PXG3
42.96
>100
17.35
>100
26.63
>100
35.05
>100
35.46
>100
20.74
>100

PXG4
0.00
>100
−7.23
>100
−10.90
>100
−5.99
>100
−7.57
>100
4.07
>100

PXG5
47.24
>100
14.08
>100
28.17
>100
23.90
>100
31.42
>100
9.57
>100

PXG6
38.85
>100
13.70
>100
15.98
>100
−1.46
>100
11.37
>100
23.21
>100

PXG7
61.50
36.12
63.59
47.19
70.09
18.3
25.89
>100
45.78
>100
46.85
>100

PXG8
38.06
>100
14.10
>100
29.11
>100
53.31
>100
35.01
>100
24.81
>100

PXG9
2.72
>100
5.71
>100
34.79
>100
−10.48
>100
5.56
>100
5.25
>100

PXG10
−2.50
>100
−3.85
>100
5.85
>100
19.24
>100
20.93
>100
9.75
>100

PXG11
31.76
>100
23.84
>100
54.79
>100
8.16
>100
20.95
>100
8.09
>100

PXG12
19.42
>100
19.48
>100
29.54
>100
−3.30
>100
12.52
>100
−5.00
>100

PXG13
40.68
>100
34.93
>100
46.18
>100
22.14
>100
19.41
>100
0.86
>100

PXG14
13.21
>100
10.91
>100
28.65
>100
−17.12
>100
9.38
>100
12.41
>100

PXG15
4.29
>100
4.69
>100
18.72
>100
2.97
>100
0.41
>100
−1.48
>100

PXG16
10.50
>100
0.41
>100
10.53
>100
−1.00
>100
−1.72
>100
2.28
>100

PXG17
−4.94
>100
12.14
>100
−5.11
>100
−16.13
>100
2.26
>100
−9.63
>100

PXG18
−2.16
>100
23.84
>100
23.00
>100
45.31
>100
28.67
>100
−3.02
>100

PXG19
47.16
>100
15.64
>100
22.03
>100
10.00
>100
19.64
>100
6.30
>100

PXG20
30.36
>100
41.65
>100
25.43
>100
−19.64
>100
32.18
>100
−2.90
>100

PXG21
18.46
>100
17.75
>100
11.87
>100
−32.48
>100
0.49
>100
5.68
>100

PXG22
60.97
19.01
42.38
>100
58.55
59.29
95.14
1.72
62.87
77.05
84.32
32.8

PXG23
22.92
>100
15.55
>100
22.83
>100
24.74
>100
22.33
>100
0.43
>100

PXG24
35.70
>100
44.66
>100
41.15
>100
42.77
>100
19.70
>100
35.93
>100

PXG25
31.23
>100
36.57
>100
51.03
>100
41.85
>100
15.80
>100
33.02
>100

PXG26
42.48
>100
−0.99
>100
17.07
>100
28.56
>100
16.01
>100
24.75
>100

PXG27
45.15
>100
46.93
>100
68.13
34.48
66.99
39.08
46.58
>100
44.63
>100

PXG28
55.24
>100
55.49
>100
58.02
60.53
45.44
>100
45.15
>100
23.64
>100

PXG29
4.90
>100
10.83
>100
13.81
>100
−10.71
>100
4.82
>100
10.99
>100

GTX
87.20
16.33
86.35
23.46
99.97
6.65
95.14
2.31
95.20
14.41
96.30
14.05

Conclusion: the compounds PXG1, PXG2 and PXG22 of the present invention have obvious inhibitory activity on the total GSTs enzymes of various field crop pests and economic crop pests, and can reduce metabolic activity towards insecticides of the total GSTs enzymes of field crop pests such as Mythimna separata, Pyrausta nubilalis, Chilo suppressalis, Nilaparvata lugens, Spodoptera frugiperda and Helicoverpa armigera, and economic crop pests such as Plutella xylostella, Carposina sasakii and Spodoptera litura.

Experimental Example 2: Enzyme Kinetic Assay of 8 Kinds of Known GSTs Recombinant Proteins in Plutella xylostella

Experiment: There are currently 8 known Plutella xylostella GSTs (PxGSTs), namely PxGSTδ1, PxGSTε3, PxGSTσ1, PxGSTσ2, PxGSTω4, PxGSTθ1, PxGSTζ1 and PxGSTμ1. The recombinant proteins of the 8 PxGSTs were obtained by prokaryotic expression, and the enzyme kinetics of the recombinant PxGSTs proteins were determined by the CDNB method. 1 μg of PxGST recombinant protein was added to 200 μL of PBS buffer (100 mM, pH 7.2) containing gradient concentrations of CDNB (0.05-1.60 mM) and 1 mM GSH. The absorbance at a wavelength of 340 nm (A₃₄₀) was measured in every 1 min at 30° C. using a microplate reader, and the measurement was continued for 5 min. The obtained light absorption value was converted into molar CDNB conjugated/min/mg, and the extinction coefficient ε340=9600 M⁻¹cm⁻¹. Michaelis-Menten plots were generated according to different concentrations of CDNB to obtain enzyme kinetic parameters.

Results: By deterring the enzyme kinetics of the recombinant protein with CDNB as substrate, it was found that only PxGSTδ1, PxGSTσ1, PxGSTσ2 and PxGSTε3 have catalytic activity on CDNB, and the remaining four PxGSTs (PxGSTω4, PxGSTθ1, PxGSTζ1 and PxGSTμ1) have no catalytic activity on CDNB. The experimental results are shown in Table 3 and FIG. 52. The K_mof the recombinant proteins of PxGSTδ1, PxGSTε3, PxGSTσ1 and PxGSTσ2 are 0.032 mM, 0.37 mM, 0.12 mM and 0.19 mM, respectively, V_maxis 7.64 mM/min/mg, 0.49 mM/min/mg, 1.70 mM/min/mg and 0.26 mM/min/mg, respectively.

TABLE 3

Kinetic parameters of Pxlutella xylostella

PxGSTs recombinant protease

PxGSTs

recombinant
K_m
V_max

protein
(mM)
(μmol min⁻¹mg⁻¹)

PxGSTδ1
0.032
7.64

PxGSTε3
0.37
0.49

PxGSTσ1
0.12
1.7

PxGSTσ2
0.19
0.26

PxGSTo4
N/A
N/A

PxGSTθ1
N/A
N/A

PxGSTu1
N/A
N/A

Conclusion: Among the 8 kinds of PxGSTs, only 4 kinds of PxGSTs of PxGSTδ1, PxGSTσ1, PxGSTσ2 and PxGSTε3 have GSH binding sites, which belong to the GST type with insecticide metabolic activity in Plutella xylostella.

Experimental Example 3: Inhibitory Activity Assay of Compound PXG1-PXG29 of the Present Invention to 4 Kinds of GSTs Having GSH Binding Site of Plutella xylostella

Experiment: 100 μL of PBS buffer (100 mM, pH 7.2) was taken and added to a 96-well plate, 1 μg of PxGSTs recombinant protein and an appropriate concentration of the compounds PXG1-PXG29 of the present invention were added to each well, after uniformly mixing, and allowed to stand at 30° C. for 10 min. Then 100 μL of PBS buffer (100 mM, pH 7.2) containing 1 mM CDNB and 1 mM GSH was successively added to each well. Immediately after mixing, the mixture was placed in a microplate reader to measure the absorbance at a wavelength of 340 nm (A₃₄₀) once every 1 min, the measurement was continued for 5 min, the light absorption values of each measuring point were recorded and the inhibition rate in vitro of the compounds PXG1-PXG29 of the present invention with different concentrations against PxGSTs recombinant protein were calculated. Each group of assays was repeated three times, and PxGSTs recombinant protein inactivated at 100° C. for 5 min was used as negative control, and GTX was used as positive control. GraphPad Prism 5 software was used for analysis and calculation of IC₅₀. Relative inhibition was calculated using the following formula:

Relative
⁢

inhibition
⁢

(
%
)

=

Δ
⁢

A

340
⁢
c

-

Δ
⁢

A

340
⁢
t

Δ
⁢

OD

340
⁢
c

×
100

- wherein, A_340cis the change of A₃₄₀within minutes or the control group, an A_340tis the change of A₃₄₀within 5 minutes for the experimental group.

Results: The experimental results are shown in Table 4 and FIG. 53 to FIG. 56. The IC₅₀of compounds PXG1, PXG2, PXG3, PXG4, PXG5, PXG7, PXG8, PXG10, PXG18, PXG20, PXG22, PXG23 and PXG28 of the present invention against the PxGSTδ1 recombinant protein of Plutella xylostella is 40.25 μM, 31.41 μM, 31.58 μM, 50.56 μM, 17.78 μM, 65.37 μM, 30.64 μM, 24.25 μM, 29.9 μM, 40.79 μM, 25.03 μM, 32.92 μM and 63.53 μM, respectively, and the IC₅₀of the other compounds of the present invention against PxGSTδ1 recombinant protein are all greater than 100 μM; the IC₅₀of compounds PXG11, PXG14, PXG15, PXG16, PXG19, PXG22, PXG26 and PXG29 of the present invention against the PxGSTσ1 recombinant protein of Plutella xylostella are 38.25 μM, 53.23 μM, 62.13 μM, 49.3 μM, 82.12 μM, 80.61 μM, 35.95 μM and 54.96 μM, respectively, and the IC₅₀of the other compounds of the present invention against PxGSTσ1 recombinant protein are all greater than 100 μM; The IC₅₀of compounds PXG1 and PXG22 of the present invention against the PxGSTσ2 recombinant protein of Plutella xylostella are 57.77 μM and 27.15 μM, respectively, and the IC₅₀of the other compounds of the present invention against PxGSTσ2 recombinant protein are all greater than 100 μM; The IC₅₀of compound PXG22 of the present invention against the PxGSTε3 recombinant protein of Plutella xylostella is 36.89 μM, and the IC₅₀of the other compounds of the present invention to PxGSTε3 recombinant protein are all greater than 100 μM. The IC₅₀of the positive control GSTs inhibitor GTX against PxGSTδ1, PxGSTσ1, PxGSTσ2 and PxGSTε3 recombinant proteins are 36.02 μM, 66.85 μM, 7.75 μM and 95.41 μM, respectively.

TABLE 4

The inhibitory effect of compounds PXG1-PXG29 of the present invention on 4 kinds of

PxGSTs recombinant protein

PxGSTδ1
PxGSTσl
PxGSTσ2
PxGSTε3

100 μM

100 μM

100 μM

100 μM

Inhibition
IC₅₀
Inhibition
IC₅₀
Inhibition
IC₅₀
Inhibition
IC₅₀

Index
rate %
(μM)
rate %
(μM)
rate %
(μM)
rate %
(μM)

PXG1
68.91
40.25
25.54
>100
58.07
57.77
17.64
>100

PXG2
87.75
31.41
25.12
>100
14.56
>100
−4.00
>100

PXG3
72.39
31.58
36.06
>100
27.59
>100
−1.11
>100

PXG4
18.41
50.56
43.63
>100
21.57
>100
8.88
>100

PXG5
81.66
17.78
35.57
>100
24.51
>100
−2.97
>100

PXG6
12.21
>100
49.08
>100
33.65
>100
4.85
>100

PXG7
55.74
65.37
35.15
>100
20.06
>100
5.93
>100

PXG8
69.47
30.64
37.78
>100
42.39
>100
−3.30
>100

PXG9
30.42
>100
28.59
>100
31.45
>100
−3.88
>100

PXG10
91.02
24.25
29.36
>100
19.17
>100
2.03
>100

PXG11
35.34
>100
60.66
38.25
22.64
>100
−7.28
>100

PXG12
40.17
>100
48.30
>100
33.65
>100
10.90
>100

PXG13
14.11
>100
34.22
>100
22.76
>100
18.43
>100

PXG14
14.93
>100
52.24
53.23
36.41
>100
15.32
>100

PXG15
7.86
>100
42.48
62.13
25.58
>100
4.62
>100

PXG16
5.35
>100
70.49
49.3
39.29
>100
4.12
>100

PXG17
2.22
>100
48.92
>100
43.85
>100
4.43
>100

PXG18
82.05
29.9
31.74
>100
13.48
>100
−2.47
>100

PXG19
58.74
40.79
45.88
82.12
32.46
>100
5.57
>100

PXG20
23.01
>100
71.80
>100
29.56
>100
−15.67
>100

PXG21
7.82
>100
44.93
>100
11.65
>100
−2.67
>100

PXG22
78.78
25.03
45.71
80.61
76.68
27.15
65.84
36.89

PXG23
77.66
32.92
35.86
>100
26.95
>100
6.04
>100

PXG24
23.09
>100
14.94
>100
19.66
>100
28.32
>100

PXG25
23.75
>100
−1.47
>100
15.27
>100
2.07
>100

PXG26
21.58
>100
67.92
35.95
24.71
>100
7.12
>100

PXG27
41.66
>100
25.10
>100
42.39
>100
17.56
>100

PXG28
70.89
63.53
19.53
>100
23.38
>100
0.96
>100

PXG29
16.73
>100
53.27
54.96
44.66
>100
0.03
>100

GTX
66.50
36.02
67.29
66.85
84.82
7.75
50.71
95.41

Conclusion: The compound PXG22 of the present invention has obvious inhibitory activity against 4 kinds of PxGSTs recombinant proteins with GSH binding sites, wherein the inhibitory activity of the compound PXG22 of the present invention against PxGSTδ1 and PxGSTε3 recombinant proteins exceeds the inhibitory activity of the positive control GSTs inhibitor GTX. The remaining compounds of the present invention have different degrees of inhibitory activity against PxGSTδ1 and PxGSTσ1 recombinant proteins, and only the compound PXG1 of the present invention has obvious inhibitory activity against PxGSTσ2 recombinant protein.

Experimental Example 4: Insecticidal Synergistic Activity Assay of the Compound PXG22 of the Present Invention with Chlorantraniliprole

Experiment: 96% chlorantraniliprole (Chl) original drug was prepared into mother liquor with a concentration of 50000 mg/L. During the test, 6 treatment solutions with concentrations of 500, 250, 125, 62.5, 31.25 and 15.63 mg/L were prepared. 200 mg/L of the compound PXG22 of the present invention was added to each treatment solution, 200 mg/L of diethyl maleate (DEM) was used as the control synergist, and water was used as another control. The Plutella xylostella (Linnaeus) larvae was collected from a vegetable field in Yunnan Province. Healthy 3rd instar larvae of the same age were selected for indoor biological activity assay.

Clean wild cabbage leaves were cut into discs with a diameter of 6.5 cm (avoiding main veins). The leaves were soaked in the pesticide solution for 10 seconds, then taken out, dried at 25° C., and placed in a petri dish with a diameter of 6.5 cm. 10 larvae of the resistant strains of Plutella xylostella at the early stage of 3rd instar were placed in each petri dish, which was covered with double-layer absorbent roll paper, and covered by the upper cover of the petri dish. It was placed face up in an incubator at a temperature of 25±1° C., a relative humidity of 65%-70%, and a light ratio (L: D) of 16: 8 h. Each treatment was replicated 4 times with 10 larvae per replicate. After 48 hours, mortality was checked, and the LC₅₀and resistance ratio were calculated. During observation, the worm body was gently touched with a small brush or sharp tweezers, if the worm body did not respond or could not coordinate movements, it was considered dead. The test process is strictly in accordance with the standard: NY/T 1154.7-2006.

Experimental data analysis was carried out using practical statistical analysis and its computer processing platform (DPS) (TANG Qiyi et al., 1997). The toxicity regression equation of each agent was established, LC₅₀value and 95% confidence limit were calculated.

Results: The experimental results are shown in Table 5. After the addition of the compound PXG22 of the present invention, the LC₅₀of the chlorantraniliprole solution to the resistant strains of Plutella xylostella decreases from 110.17 mg/L to 14.04 mg/L, and the resistance ratio of resistant strains of Plutella xylostella to chlorantraniliprole falls from 479 to 61.04. For the chlorantraniliprole solution added with the control synergist DEM, its LC₅₀and resistance ratio decreases to 50.02 mg/L and 217.48, respectively.

TABLE 5

Synergistic effect of compounds of the present invention and DEM with chlorantraniliprole

Correlation

treatment
regression
LC₅₀
confidence
coefficient
Resistance

group
equation
(mg/L)
interval
R²
ratio

Chl
Y = 0.95X − 1.94
110.17
67.72-172.48
0.927
479

Chl + DEM
Y = 1.15X − 1.95
50.02
34.53-75.11
0.984
217.48

Chl + PXG22
Y = 1.75X − 2.01
14.04
0.45-22.73
0.881
61.04

Note:

Chl: Chlorantraniliprole

Conclusion: The compound PXG22 of the present invention has obvious insecticidal synergistic activity with chlorantraniliprole, and its synergistic activity exceeds that of the control synergist DEM.

Experimental Example 5: Insecticidal Synergistic Activity Assay of the Compounds PXG1, PXG2 and PXG22 of the Present Invention with Indoxacarb

Experiment: 95% indoxacarb (Ind) original drug was prepared into mother liquor with a concentration of 50000 mg/L. During the test, 6 treatment solutions with concentrations of 500, 250, 125, 62.5, 31.25 and 15.63 mg/L were prepared. 200 mg/L of the compounds PXG1, PXG2 and PXG22 of the present invention were added to each treatment solution, 200 mg/L of diethyl maleate (DEM) was used as the control synergist, and water was used as another control. The Plutella xylostella (Linnaeus) larvae was collected from a vegetable field in Huizhou city, Guangdong Province. Healthy 3rd instar larvae of the same age were selected for indoor biological activity assay.

Clean wild cabbage leaves were cut into discs with a diameter of 6.5 cm (avoiding main veins). The leaves were soaked in the pesticide solution for 10 seconds, then taken out, dried at 25° C., and placed in a petri dish with a diameter of 6.5 cm. 10 larvae of the resistant strains of Plutella xylostella at the early stage of 3rd instar were placed in each petri dish, which was covered with double-layer absorbent roll paper, and covered by the upper cover of the petri dish. It was placed face up in an incubator at a temperature of 25±1° C., a relative humidity of 65%-70%, and a light ratio (L: D) of 16: 8 h. Each treatment was replicated 4 times with 10 larvae per replicate. After 48 hours, mortality was checked, and the LC₅₀and resistance ratio were calculated. During observation, the worm body was gently touched with a small brush or sharp tweezers, if the worm body did not respond or could not coordinate movements, it was considered dead. The test process is strictly in accordance with the standard: NY/T 1154.7-2006.

Experimental data analysis was carried out using practical statistical analysis and its computer processing platform (DPS) (TANG Qiyi et al., 1997). The toxicity regression equation of each agent was established, LC₅₀value and 95% confidence limit were calculated.

Results: The experimental results are shown in Table 6. After the addition of the compound PXG1 of the present invention, the LC₅₀of the indoxacarb to the resistant strains of Plutella xylostella decreases from 74.72 mg/L to 59.13 mg/L, and the resistance ratio of resistant strains of Plutella xylostella to indoxacarb falls from 143.70 to 113.71; After the addition of the compound PXG2 of the present invention, the LC₅₀of the indoxacarb to the resistant strains of Plutella xylostella decreases from 74.72 mg/L to 39.93 mg/L, and the resistance ratio of resistant strains of Plutella xylostella to indoxacarb falls from 143.70 to 76.78; After the addition of the compound PXG22 of the present invention, the LC₅₀of the indoxacarb to the resistant strains of Plutella xylostella decreases from 74.72 mg/L to 51.21 mg/L, and the resistance ratio of resistant strains of Plutella xylostella to indoxacarb falls from 143.70 to 98.48; For the indoxacarb solution added with the control synergist DEM, its LC₅₀and resistance ratio decrease to 54.11 mg/L and 104.06, respectively.

TABLE 6

Synergistic effect of compounds of the present invention and DEM with indoxacarb

Correlation

treatment
regression
LC₅₀
confidence
coefficient
Resistance

group
equation
(mg/L)
interval
R²
ratio

Ind
Y = 1.32X − 2.47
74.72
48.82-106.92
0.860
143.70

Ind + DEM
Y = 1.16X − 2.01
54.11
29.03-79.18
0.903
104.06

Ind + PXG1
Y = 1.111X − 1.97
59.13
32.67-86.20
0.955
113.71

Ind + PXG2
Y = 1.17X − 1.88
39.93
18.05-61.39
0.997
76.78

Ind + PXG22
Y = 1.14X − 1.94
51.21
26.43-76.19
0.948
98.48

Note:

Ind: Indoxacarb

Conclusion: tresistance ratiohe compounds PX1, PXG2 and PXG22 of the present invention have obvious synergistic activity with indoxacarb, and PXG2 and PXG22 have significant effects, which can make the resistance of resistant strains of Plutella xylostella to indoxacarb from high resistance to moderate resistance.

Experimental Example 6: Insecticidal Synergistic Activity Assay of the Compound PXG22 of the Present Invention with Chlorantraniliprole Using the Ultra-High Resistant Strains of Plutella xylostella (Huizhou Insect Source) as the Tested Insect

Experiment: The method was the same as in Experimental Example 4. The tested Plutella xylostella was ultra-high resistance strains to chlorantraniliprole (Huizhou insect source). The compound PXG22 of the present invention was added at a concentration of 200 mg/L. The control synergist diethyl maleate (DEM) was added at a concentration of 200 mg/L.

Results: The experimental results are shown in Table 7. After the addition of compound PXG22 of the present invention, the LC₅₀of the chlorantraniliprole solution to the ultra-high resistant strains of Plutella xylostella (Huizhou insect source) decreases from 503.38 mg/L to 148.45 mg/L, and the resistance ratio of ultra-high resistant strains of Plutella xylostella (Huizhou insect source) to chlorantraniliprole falls from 2188.61 to 645.43. For the chlorantraniliprole solution added with the control synergist DEM, its LC₅₀and resistance ratio decrease to 442.81 mg/L and 1925.26, respectively.

TABLE 7

Synergistic effect of compound PXG22 of the present invention and DEM with chlorantraniliprole

Correlation

treatment
regression
LC₅₀
confidence
coefficient
Resistance

group
equation
(mg/L)
interval
R²
ratio

Chl
Y = 2.39X − 6.46
503.38
375.19-719.83
0.860
2188.61

Chl + DEM
Y = 2.87X − 7.86
442.81
369.82-540.47
0.903
1925.26

Chl + PXG22
Y = 1.53X − 3.33
148.45
72.60-235.24
0.918
645.43

Note:

Chl: Chlorantraniliprole

Conclusion: The compound PXG22 of the present invention significantly enhances the insecticidal activity of chlorantraniliprole to ultra-high resistant strains of Plutella xylostella (Huizhou insect source), and its synergistic activity with chlorantraniliprole is significantly higher than that of the control synergist DEM.

Experimental Example 7: Insecticidal Synergistic Activity Assay of the Compound PXG22 of the Present Invention Having Different Concentrations with Chlorantraniliprole Using the High Resistant Strains I of Plutella xylostella (Lianzhou Insect Source) as the Tested Insect

Experiment: The method was the same as in Experimental example 4, and the tested Plutella xylostella was the high resistant strains I against chlorantraniliprole (Lianzhou source). The added concentrations of the compound PXG22 of the present invention were 200 mg/L, 100 mg/L and 50 mg/L, and the added concentration of diethyl maleate (DEM) of the control synergist was maintained at 200 mg/L.

Results: The experimental results are shown in Table 8. After the addition of 200 mg/L, 100 mg/L and 50 mg/L of the compound PXG22 of the present invention, the LC₅₀of the chlorantraniliprole solution to the high resistant strains I of Plutella xylostella (Lianzhou insect source) decreases from 134.26 mg/L to 40.44 mg/L, 21.16 mg/L and 49.89 mg/L, respectively, and the resistance ratio of high resistant strains I of Plutella xylostella (Lianzhou insect source) to chlorantraniliprole falls from 583.74 to 175.82, 92 and 216.91 respectively. For the chlorantraniliprole solution added with the control synergist DEM, its LC₅₀and resistance ratio decrease to 53.36 mg/L and 232, respectively.

TABLE 8

Synergistic effect of compound PXG22 of the present invention and

DEM with chlorantraniliprole

Correlation

regression
LC₅₀
confidence
coefficient
Resistance

treatment group
equation
(mg/L)
interval
R²
ratio

Chl
Y = 0.99X − 2.10
134.26
69.30-208.45
0.983
583.74

Chl + 200 mg/L PXG22
Y = 1.26X − 2.03
40.44
6.45-75.75
0.989
175.82

Chl + 100 mg/L PXG22
Y = 1.04X − 1.38
21.16
8.37-33.80
0.979
92

Chl + 50 mg/L PXG22
Y = 1.30X − 2.20
49.89
28.11-71.00
0.901
216.91

Chl + DEM
Y = 0.79X − 1.36
53.36
16.57-95.03
0.967
232

Note:

Chl: Chlorantraniliprole

Conclusion: The compound PXG22 of the present invention significantly enhances the insecticidal activity of chlorantraniliprole against high resistant strains I of Plutella xylostella (Lianzhou insect source), and when the compound PXG22 of the present invention is added with a concentration of 100 mg/L, its synergistic activity with chlorantraniliprole is the strongest, which is significantly higher than that of the control synergist DEM.

Experimental Example 8: Insecticidal Synergistic Activity Assay of the Compound PXG22 of the Present Invention Having Different Concentrations with Chlorantraniliprole Using the High Resistant Strains II of Plutella xylostella (Lianzhou Insect Source) as the Tested Insect

Experiment: The method was the same as in Experimental example 4, and the tested Plutella xylostella was the high resistant strains II against chlorantraniliprole (Lianzhou source). The added concentrations of the compound PXG22 of the present invention were 200 mg/L, 100 mg/L, 50 mg/L and 25 mg/L, and the added concentration of diethyl maleate (DEM) of the control synergist was maintained at 200 mg/L.

Results: The experimental results are shown in Table 9. After the addition of 200 mg/L, 100 mg/L, 50 mg/L and 25 mg/L of the compound PXG22 of the present invention, the LC₅₀of the chlorantraniliprole solution to the high resistant strains II of Plutella xylostella (Lianzhou insect source) decrease from 145.19 mg/L to 55.92 mg/L, 44.86 mg/L, 71.88 mg/L and 78.54 mg/L, respectively, and the resistance ratio of high resistant strains II of Plutella xylostella (Lianzhou insect source) to chlorantraniliprole fall from 631.26 to 243.13, 195.04, 312.52 and 341.48 respectively. For the chlorantraniliprole solution added with the control synergist DEM, its LC₅₀and resistance ratio decrease to 107.99 mg/L and 469.52, respectively.

TABLE 9

Synergistic effect of compound PXG22 of the present invention and

DEM with chlorantraniliprole

Correlation

regression
LC₅₀
confidence
coefficient
Resistance

treatment group
equation
(mg/L)
interval
R²
ratio

Chl
Y = 1.54X − 3.33
145.19
111.97-192.13
0.940
631.26

Chl + 200 mg/L PXG22
Y = 1.67X − 2.91
55.92
38.60-73.32
0.964
243.13

Chl + 100 mg/L PXG22
Y = 1.56X − 2.58
44.86
27.78-61.46
0.984
195.04

Chl + 50 mg/L PXG22
Y = 1.76X − 3.26
71.88
48.99-95.51
0.993
312.52

Chl + 25 mg/L PXG22
Y = 1.31X − 2.47
78.54
48.23-111.47
0.958
341.48

Chl + DEM
Y = 1.49x − 3.032
107.99
82.46-140.86
0.932
469.52

Note:

Chl: Chlorantraniliprole

Conclusion: The compound PXG22 of the present invention significantly enhances the insecticidal activity of chlorantraniliprole against high resistant strains II of Plutella xylostella (Lianzhou insect source), and when the compound PXG22 of the present invention is added with a concentration of 100 mg/L, its synergistic activity with chlorantraniliprole is the strongest, which is significantly higher than that of the control synergist DEM.

Based on the above description of the summary of the invention, those skilled in the art will be able fully apply the present invention, and all the same principles or similar modifications should be considered to be within the scope of the present invention.