Derivative of targeted squalene epoxidase as well as preparation method and application thereof

Abstract

The invention relates to a derivative of targeted squalene epoxidase as well as a preparation method and application thereof, in particular to application of thiazole imine-containing compounds or agrochemically acceptable salts thereof in prevention and treatment of plant fungal diseases, and part of the compounds show relatively good agricultural bactericidal activity and can be used as potential commercial products.

Description

Technical Field

[0001] This invention belongs to the field of pesticide chemistry, specifically relating to a derivative targeting squalene epoxidase, its preparation method, and its uses. Background Technology

[0002] The global population is projected to reach 9.8 billion by 2050, leading to a surge in food demand. Meanwhile, pesticide resistance caused by overuse poses a serious threat to agricultural production. Developing pesticides with novel mechanisms of action is a cost-effective integrated pest management solution.

[0003] Squalene epoxidase (SQLE) belongs to the flavoprotein monooxygenase family and is one of the rate-limiting enzymes in cholesterol synthesis. It primarily catalyzes the oxidation of squalene to 2,3(S)-oxidized squalene. Squalene epoxidase is a validated and crucial target in pesticide chemistry. Fungicides and acaricides developed based on this target play an irreplaceable role in modern agricultural disease management.

[0004] Therefore, developing more pesticide derivatives that target squalene epoxidase is an urgent agricultural problem that needs to be solved. Summary of the Invention

[0005] The object of this invention is to provide the application of compounds of the following formula, or stereoisomers thereof, or agriculturally chemically acceptable salts thereof, in the control of plant fungal diseases. ; Preferably, the plant fungus is selected from one or more of the following: early blight pathogen of tomato, Botrytis cinerea, brown spot pathogen of peanut, Fusarium graminearum, apple ring rot pathogen, Rhizoctonia solani, Sclerotinia sclerotiorum, Fusarium scutellaria, and Fusarium verticillata.

[0006] A further preferred embodiment of the present invention: the compound, or its stereoisomer, or its agriculturally chemically acceptable salt is the active ingredient, and the mass percentage of the active ingredient is from 0.1% to 99.9%.

[0007] Furthermore, the solid or liquid additives comprise 99.9% to 0.1% by mass, and optionally 0% to 25% by mass of surfactant.

[0008] The present invention also includes a pesticide composition comprising at least one compound as described above, or a stereoisomer thereof, or an agriculturally chemically acceptable salt thereof, and a carrier thereof; said carrier being in liquid or solid form.

[0009] The present invention further includes a bactericide containing a bactericidally effective amount of at least one of the compounds described above, or stereoisomers thereof, or agriculturally chemically acceptable salts thereof, and optionally containing excipients.

[0010] More preferably, the formulation of the bactericide is selected from at least one of emulsifiable concentrate, suspension concentrate, wettable powder, powder, granule, aqueous solution, mother liquor, and masterbatch.

[0011] The present invention also includes a method for controlling or preventing the infection of useful plants by plant pathogenic microorganisms, wherein an effective amount of a fungicide according to the compound as described above or a composition containing such compound as an active ingredient is applied to the plant, its parts or the site thereof.

[0012] The following is an explanation and description of the terminology used in this invention: The term "agronomically acceptable salt" refers to acidic and / or basic salts formed by the above-mentioned compounds or their stereoisomers with inorganic and / or organic acids and bases, including zwitterionic salts (internal salts) and quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly during the final separation and purification of the compounds. Alternatively, they can be obtained by mixing the above-mentioned compounds, or their stereoisomers, with an appropriate amount (e.g., equimolar amounts) of an acid or base. These salts may be obtained by precipitating in solution and collecting by filtration, by recovery after solvent evaporation, or by freeze-drying after reaction in an aqueous medium. The salts described in this invention can be hydrochlorides, sulfates, citrates, benzenesulfonates, hydrobromides, hydrofluoric acids, phosphates, acetates, propionates, succinates, oxalates, malates, succinates, fumarates, maleates, tartrates, or trifluoroacetates of the compounds.

[0013] The compounds of this invention are used in related fields as active ingredients, for example, for controlling plant pests, or on non-living materials for controlling putrefactive microorganisms or potentially harmful organisms to humans. These novel compounds of the invention are characterized by excellent activity at low application rates, good plant tolerance, and no environmental harm. They have very useful therapeutic, preventative, and systemic properties and can be used to protect cultivated plants. The compounds of the invention can be used to inhibit or destroy pests appearing on various useful plant crops or plant parts (fruits, flowers, leaves, stems, tubers, roots), while also protecting, for example, later-growing plant parts from plant pathogenic microorganisms.

[0014] The term "plant" refers to all the tangible parts of a plant, including seeds, seedlings, saplings, roots, tubers, stems, culms, leaves, and fruits.

[0015] The compound of the present invention may be the sole active ingredient in a composition, or, where appropriate, it may be mixed with one or more other active ingredients (such as pest control agents, fungicides, synergists, herbicides, or plant growth regulators). In some cases, the additional active ingredients may result in unexpected synergistic activity.

[0016] Other suitable active ingredients include acyclic amino acid fungicides, aliphatic nitrogen fungicides, amide fungicides, aniline fungicides, and antibiotic fungicides.

[0017] A formulation, such as a composition comprising the compound of the present invention, and a solid or liquid adjuvant or a monomer for encapsulating the compound of the present invention, can typically be prepared in a known manner by closely mixing and / or grinding the compound with an additive (e.g., a solvent, a solid carrier, and, optionally, a surfactant compound).

[0018] Typically, formulations comprise from 0.01% to 90% by weight of the active ingredient, from 0% to 20% of an agriculturally acceptable surfactant, and from 10% to 99.99% of solid or liquid formulation inert agents and adjuvants.

[0019] The compositions of the present invention can be used in any conventional form, for example, as a double-pack, emulsion concentrate (EC), suspension concentrate (SC), suspension emulsion (SE), capsule suspension (CS), water-dispersible particles (WG), emulsifiable particles (EG), water-in-oil emulsion (EO), oil-in-water emulsion (EW), microemulsion (ME), dispersible oil suspension (OD), oil suspension (OF), oil-soluble liquid (OL), soluble concentrate (SL), ultra-low volume suspension (SU), ultra-low volume liquid (UL), technical grade (TK), dispersible concentrate (DC), wettable powder (WP), or any technically feasible formulation in combination with agriculturally acceptable adjuvants.

[0020] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.

[0021] The beneficial technical effects of the present invention are as follows: 1. The present invention provides a class of thiazolidinylamine derivatives; the derivatives of the present invention can regulate the biological activity of agricultural, horticultural and health and forestry plant pests and plant pathogens, and can be used in the fields of agriculture, horticulture and forestry for insecticidal, acaricidal, bactericidal, antiviral and induction of plant disease resistance, and have good economic value and application prospects.

[0022] 2. Compounds gw-76, ytf-34, and ytf-36 exhibit good agricultural fungicidal activity and can be considered as potential commercial products. Attached Figure Description

[0023] Figure 1 : Inhibitory activity of compound ytf-34 of the present invention against Rhizoctonia solani at a concentration of 50 μg / mL; Figure 2 : Inhibitory activity of the compound ytf-36 against Rhizoctonia solani at a concentration of 50 μg / mL. Detailed Implementation

[0024] The present invention is further illustrated by the following examples, but these are not intended to limit the invention.

[0025] The preparation method of the present invention adopts the following general formula preparation method: ; Where R 1 R 2 The corresponding groups correspond to the compounds in the intermediate examples and the examples, respectively.

[0026] Intermediate Example 1: Preparation of Compound Ib Phenyl isothiocyanate (18.0 mmol), ammonia (36 mmol), and 40 mL of ethanol were added to a 100 mL round-bottom flask. The mixture was then stirred at 80 °C for 6 hours. The reaction progress was monitored by thin-layer chromatography (TLC). After the reaction was complete, the reaction solvent was removed under reduced pressure, and the crude product was crystallized from ethanol to give intermediate Ib, a white solid, in 92% yield.

[0027] Example 1: Preparation of compound gw-51 Intermediate Ib (2.0 mmol) was added to a 50 mL round-bottom flask along with 1-cyclopropyl-2-bromoethylone (2.0 mmol) and 20 mL of ethanol, and the mixture was reacted at 80°C for 4 hours. After the reaction was complete, the reaction mixture was evaporated to dryness, dissolved in dichloromethane, and extracted twice (50 mL × 2) with saturated sodium bicarbonate solution. The aqueous phase was then extracted twice (50 mL × 2) with dichloromethane. The combined organic layers were washed twice with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The residue was filtered under reduced pressure and then treated with petroleum ether / ethyl acetate (1:1 to 3:1) with a boiling range of 60–90°C. yes / yes Using 100-200 mesh silica gel column chromatography as the eluent, the target compound gw-51 was purified as a white solid with a yield of 87%.

[0028] 1H NMR (500 MHz, CDCl3) δ 11.50 (s, 1H), 7.46 (dd, J = 8.3, 7.6 Hz,2H), 7.38 – 7.30 (m, 3H), 6.14 (s, 1H), 2.00 – 1.91 (m, 1H), 1.11 (dt, J =7.0, 5.0 Hz, 2H), 1.05 – 0.98 (m, 2H). 13 C NMR (126 MHz, CDCl3) δ 167.65 (s), 143.85 (s), 136.50 (s), 130.19 (s), 127.45 (s), 121.09 (s), 97.43 (s), 9.45 (s), 8.05 (s). Example 2: Preparation of compound GW-76 Following the preparation steps of Example 1, compound GW-76 was obtained. 1 H NMR (500 MHz, CDCl3) δ 9.17(s, 1H), 7.34 – 7.30 (m, 2H), 7.28 – 7.23 (m, 2H), 6.16 (d, J = 0.4 Hz, 1H), 2.86 (dt, J = 13.6, 6.8 Hz, 1H), 1.24 (d, J = 6.9 Hz, 6H). 13 C NMR (126 MHz, CDCl3) δ 165.04 (s), 157.03 (s), 138.57 (s), 129.46 (s), 128.39 (s), 119.97 (s), 99.49 (s), 30.37 (s), 21.86 (s). Example 3: Preparation of compound GW-77 Following the preparation steps of Example 1, compound GW-77 was obtained. 1 H NMR (500 MHz, CDCl3) δ 9.70(s, 1H), 7.33 (d, J = 9.0 Hz, 2H), 7.31 – 7.27 (m, 2H), 6.20 (s, 1H), 2.64(dd, J = 14.5, 7.2 Hz, 2H), 1.26 (t, J= 7.4 Hz, 3H). Example 4: Preparation of compound GW-78 Following the preparation steps of Example 1, compound GW-78 was obtained. 1 H NMR (500 MHz, CDCl3) δ 8.37(s, 1H), 7.29 – 7.22 (m, 2H), 7.21 – 7.15 (m, 2H), 6.11 (s, 1H), 1.23 (s,9H). 13 C NMR (126 MHz, CDCl3) δ 163.83 (s), 161.12 (s), 138.79 (s), 129.39 (s), 127.81 (s), 119.34 (s), 99.33 (s), 34.63 (s), 29.67 (s). Example 5: Preparation of compound GW-91 Following the preparation steps of Example 1, compound GW-91 was obtained. 1 H NMR (500 MHz, CDCl3) δ 7.29(t, J = 7.4 Hz, 2H), 7.21 (t, J = 8.6 Hz, 3H), 6.06 (s, 1H), 5.35 (s, 1H), 3.45(t, J = 6.6 Hz, 2H), 2.91 (t, J = 7.2 Hz, 2H), 1.26 (s, 9H). 13 C NMR (126 MHz, CDCl3) δ 169.19 (s), 162.96 (s), 138.65 (s), 128.85 (s), 128.75 (s), 126.66 (s), 97.90 (s), 47.36 (s), 35.60 (s), 34.66 (s), 29.80 (s). Example 6: Preparation of compound GW-92 Following the preparation steps of Example 1, compound GW-92 was obtained. 1 H NMR (500 MHz, DMSO) δ 10.33(s, 1H), 7.97 (t, J = 1.8 Hz, 1H), 7.46 (dd, J = 8.2, 1.3 Hz, 1H), 7.31 (t, J=8.1 Hz, 1H), 6.96 (dd, J = 7.9, 1.2 Hz, 1H), 6.47 (s, 1H), 1.29 (s, 9H). 13 C NMR (126 MHz, DMSO) δ 162.41 (s), 162.05 (s), 143.28 (s), 133.80 (s), 130.89 (s), 120.67 (s), 116.44 (s), 115.39 (s), 100.50 (s), 34.87 (s), 30.05 (s). Example 7: Preparation of compound GW-94 Following the preparation steps of Example 1, compound GW-94 was obtained. 1 H NMR (500 MHz, DMSO) δ 9.62(s, 1H), 8.49 (d, J = 8.4 Hz, 1H), 7.58 (s, 1H), 7.40 (d, J = 8.2 Hz, 1H), 6.51(s, 1H), 1.26 (s, 9H). 13 C NMR (126 MHz, DMSO) δ 162.82 (s), 161.54 (s), 137.56 (s), 129.21 (s), 128.11 (s), 125.61 (s), 122.59 (s), 121.36 (s), 101.94 (s),34.80 (s), 30.07 (s). Example 8: Preparation of compound GW-98 Following the preparation steps of Example 1, compound GW-98 was obtained. 1 H NMR (500 MHz, CDCl3) δ 8.07 –8.02 (m, 1H), 7.84 (dd, J = 6.4, 2.8 Hz, 1H), 7.75 (d, J = 7.5 Hz, 1H), 7.64(d, J = 8.2 Hz, 1H), 7.50 – 7.45 (m, 2H), 7.45 – 7.40 (m, 1H), 6.13 (s, 1H), 1.30 (s, 9H). 13C NMR (126 MHz, CDCl3) δ 166.93 (s), 161.94 (s), 136.40 (s), 134.53 (s), 128.64 (s), 127.32 (s), 126.48 (s), 126.38 (s), 125.89 (s),125.14 (s), 121.36 (s), 117.72 (s), 99.17 (s), 34.62 (s), 29.80 (s). Example 9: Preparation of compound YTF-34 Following the preparation steps of Example 1, compound YTF-34 was obtained. 1 H NMR (500 MHz, CDCl3) δ 11.44(s, 1H), 7.46 (dd, J = 8.3, 7.6 Hz, 2H), 7.36 – 7.30 (m, 3H), 6.22 (s, 1H), 1.42 (s, 9H). 13 C NMR (126 MHz, CDCl3) δ 168.15 (s), 150.76 (s), 136.55 (s), 130.18 (s), 127.37 (s), 121.03 (s), 97.48 (s), 33.79 (s), 29.05 (s). Example 10 Preparation of compound YTF-36 Following the preparation steps of Example 1, compound YTF-36 was obtained. 1 H NMR (500 MHz, CDCl3) δ 11.45(s, 1H), 7.46 (t, J = 7.7 Hz, 2H), 7.37 – 7.32 (m, 3H), 6.26 (s, 1H), 3.02(dq, J = 13.8, 6.8 Hz, 1H), 1.38 (d, J = 7.0 Hz, 6H). 13 C NMR (126 MHz, CDCl3) δ168.03 (s), 147.42 (s), 136.53 (s), 130.18 (s), 127.38 (s), 121.01 (s), 98.19(s), 28.47 (s), 21.17 (s). Example 11 Preparation of compound YTF-43 Following the preparation steps of Example 1, compound YTF-43 was obtained. 1 H NMR (500 MHz, CDCl3) δ 8.06(dd, J = 8.2, 1.2 Hz, 1H), 7.72 (s, 1H), 7.37 (dd, J = 8.0, 1.4 Hz, 1H), 7.29 –7.21 (m, 1H), 6.94 (td, J = 7.8, 1.3 Hz, 1H), 6.25 (s, 1H), 2.95 (dt, J = 13.7, 6.8 Hz, 1H), 1.29 (d, J = 6.9 Hz, 6H). 13 C NMR (126 MHz, CDCl3) δ 162.92 (s),159.65 (s), 137.16 (s), 129.52 (s), 127.87 (s), 122.70 (s), 121.85 (s),117.81 (s), 100.77 (s), 30.97 (s), 22.06 (s). Example 12 Preparation of compound YTF-48 Following the preparation steps of Example 1, compound YTF-48 was obtained. 1 H NMR (500 MHz, CDCl3) δ 9.94(s, 1H), 7.45 – 7.40 (m, 2H), 7.37 (d, J = 7.5 Hz, 2H), 7.24 (dd, J = 10.4, 4.2Hz, 1H), 6.24 (s, 1H), 2.35 (s, 3H). 13 C NMR (126 MHz, CDCl3) δ 167.19 (s), 139.50 (s), 137.49 (s), 129.99 (s), 126.40 (s), 120.54 (s), 100.71 (s), 14.71 (s). Example 1 The bactericidal or bacteriostatic activity of the thiazole imine derivatives of this invention is determined by the mycelial growth rate method. This method utilizes the rate of growth of pathogenic mycelia on culture media containing different concentrations of the agent to determine the bactericidal toxicity. The specific steps are as follows: (1) Preparation of drug-loaded plates. Take 10 mg of the test compound and use 400 μL of... N , N Dissolve the reagent in dimethylformamide and bring the volume to 20 mL with a sterile aqueous solution containing 0.1% Tween 80 emulsifier, obtaining a test reagent solution with a concentration of 500 μg / mL. Prepare the culture medium containing the reagent in a laminar flow hood. Pipette 4 mL of the test reagent solution into a sterile 50 mL centrifuge tube, add 36 mL of potato dextrose agar (PDA) medium, mix well, pour into sterile petri dishes, and after cooling, obtain a culture medium plate containing 50 μg / mL of the test reagent. Add the appropriate volume... N , N - Dimethylformamide and PDA medium containing 0.1% Tween 80 in sterile water were used as controls.

[0029] (2) Prepare the mycelium. Inoculate the long-term stored mycelium onto drug-free PDA medium and activate it 1-2 times. After the mycelium grows well, use a 4 mm diameter punch to cut the mycelium at the outer 1 / 3 of the colony.

[0030] (3) Inoculation of mycelial cakes. Use an inoculation needle to transfer the mycelial cakes to the center of the drug-coated plate, with the aerial mycelial side facing down. After sealing with sealing film, place the plate in an inverted incubator at a constant temperature of 24±1℃. Each treatment is replicated 3 times.

[0031] (4) Data collection. When the colonies of the blank control were close to the edge of the petri dish, the colony diameter was measured twice using the "cross-cross method", and the average value was used to represent the colony size.

[0032] (5) Data Analysis. Substitute the data into the growth rate calculation formula to calculate the mycelial growth inhibition rate of different compounds. The tested strains represent most of the typical plant pathogens actually occurring in the field in my country's agricultural production; their codes and names are as follows: As The Latin name of the tomato early blight pathogen is: Alternaria solani , Bc The fungus causing gray mold in cucumbers has the Latin name: Botrytis cinerea , Ca The fungus causing brown spot in peanuts has the Latin name: Cercospora arachidicola , Fg Fusarium graminearum, the causal agent of the disease, has the Latin name: Fusarium gramineae , Pp Apple ring rot fungus, its Latin name is: Physalospora piricola , Rs. The fungus causing rice sheath blight has the Latin name: Rhizoctonia solani Ss: Sclerotinia sclerotiorum, the causal agent of rapeseed disease, has the Latin name: Sclerotinia sclerotiorum Fusarium scutellatus, Fusarium oxysporum ( F. o ) and Fusarium pseudoverticum, Fusarium verticillium ( F. v ) 。

[0033] The formula for calculating the mycelial growth inhibition rate is as follows: ; Table 1. Antibacterial activity of the compounds of the present invention (inhibition rate at 50 μg / mL / %) ; Note: The pathogen causing early blight of tomatoes. A. solani ( A. s Botrytis cinerea, B. cinerea ( B. c Peanut brown spot pathogen, C. arachidicola ( C. a Fusarium graminearum, F. graminearum ( F. g Apple ring rot pathogen, P. piricola ( P. p Rhizoctonia solani, R. solani ( R.s ); Sclerotium sclerotiorum, S. sclerotiorum ( S. s Fusarium scutellatus, Fusarium oxysporum ( F. o ); Fusarium pseudoverticum, Fusarium verticillium ( F. v ).

[0034] Commercially available agents thifluzamide and triamcinolone were used as positive controls. Table 1 shows the in vitro bactericidal activity at a concentration of 50 μg / mL, indicating that the target compound in this invention exhibits a certain degree of in vitro bactericidal activity against all nine tested pathogenic fungi.

[0035] Compound GW-78 A. solani The inhibition rate was 65.1±0%, showing antibacterial activity comparable to thifluzamide (67.2%).

[0036] Compounds GW-78, YTF-34, YTF-36, and YTF-43 all exhibited inhibition rates greater than 70% against *B. cinerea*, superior to thifluzamide (32.4%). Among them, compounds GW-76, GW-78, and YTF-36 showed inhibition rates of 94.3%, 92.2%, and 95.4%, respectively, comparable to fenbendazole (96.1%). Compound YTF-34... F. graminearum The inhibition rate was 95.4%, which was superior to thifluzamide (43.6%). The series of compounds of this invention showed [resistance to...]. R. solani They exhibit excellent inhibitory activity. Compounds GW-76, YTF-34, YTF-36, and YTF-43 showed excellent inhibitory activity. R. solani The inhibitory effect was greater than 90%, showing superior or comparable inhibitory activity compared to or better than that of triamcinolone (95.3%) and thifluzamide (90.2%).

[0037] Therefore, compounds gw-76, gw-78, ytf-34, ytf-36, and ytf-43 exhibited good agricultural fungicidal activity.

[0038] The above description is merely a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. The application of the compound shown in the following formula, or its stereoisomer, or its agrochemically acceptable salt, in the control of plant fungal diseases. 。 2. The application according to claim 1, characterized in that: The plant fungi are selected from one or more of the following: early blight pathogens of tomato, Botrytis cinerea, brown spot pathogens of peanut, Fusarium graminearum, apple ring rot pathogens, Rhizoctonia solani, Sclerotinia sclerotiorum, Fusarium scutellaria, and Fusarium verticillata.

3. The application according to any one of claims 1-2, characterized in that: The compound, or its stereoisomer, or its agriculturally acceptable salt, is the active ingredient, and the mass percentage of the active ingredient is from 0.1% to 99.9%.

4. The application according to claim 3, characterized in that: The solid or liquid additives are present in a mass percentage of 99.9% to 0.1%, and optionally in a mass percentage of 0 to 25% surfactants.

5. A pesticide composition comprising at least one compound of claim 1, or a stereoisomer thereof, or an agriculturally chemically acceptable salt thereof, and a carrier thereof; said carrier being in liquid or solid form.

6. A bactericide, characterized in that, The disinfectant contains at least one of the following: a bactericidal effective amount of the compound of claim 1, or a stereoisomer thereof, or an agriculturally chemically acceptable salt thereof, and optionally contains excipients.

7. The bactericide according to claim 6, characterized in that, The formulation of the bactericide is selected from at least one of emulsifiable concentrate, suspension concentrate, wettable powder, powder, granule, aqueous solution, mother liquor, and masterbatch.

8. A method for controlling or preventing the infection of useful plants by plant pathogenic microorganisms, wherein an effective amount of the fungicide according to claim 1 or a composition comprising such compound as an active ingredient is applied to the plant, its parts or the site thereof.

Images