Derivative of isoxazoline compound and use thereof

Abstract

The application provides a derivative of an isoxazoline compound and application thereof in preventing and treating parasitic infections, and a structural general formula of the derivative is as follows: The derivative has obvious inhibiting and killing effects on most in-vitro parasites, and is suitable for preparing animal anti-parasite drugs. The derivative has high safety, small irritation, good curative effect and obvious target effect, and has wide application prospects in preventing and treating in-vitro parasites and external environment parasites of animals and agricultural pests.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical compounds, and particularly relates to a derivative of an isoxazoline compound and application thereof. BACKGROUND

[0002] Mammals, domestic animals and the like are often subject to infestation by parasites. Domestic animals such as cats and dogs are often infested with one or more of the following ectoparasites: cat and dog fleas (Ctenocephalides felis, Ctenocephalides spp., etc.), ticks (Rhipicephalus spp., Ixodes spp., Dermacentor spp., etc.), and mites (Demodex spp., Sarcoptes spp., Otodectes spp., etc.), lice (Trichodectes spp., Lingenfeldera spp., etc.), mosquitoes (Aedes spp., Culex spp., Anopheles spp., etc.) and flies (Musca spp., Stomoxys spp., Dermatobia spp., Ophyra spp., etc.). Prevalent parasites in domestic animals are ticks of the genus Boophilus, particularly the species B. microplus (cattle tick) and B. annulatus. Ticks, such as B. microplus, are particularly difficult to control because they live in the pastures where the domestic animals graze. Other important parasites of cattle and sheep, listed in decreasing order of importance, are flies that cause myiasis such as Dermatobia hominis and Ophyra phoebicola; flies that cause flystrike in sheep such as Lucilia sericata and Calliphora erythrocephala. These are flies whose larvae constitute animal parasites; dipterans, i.e. those whose adult form constitutes the parasite, such as Haematobia irritans; lice such as Linognathus vitulorum; and mites such as Psoroptes ovis.

[0003] Therefore, the control of parasitic infections in animal populations has always been an important global task. Isoxazoline drugs are a class of broad-spectrum insecticides that can inhibit L-glutamate and γ-aminobutyric acid ligand-gated chloride channels at different positions, which are ubiquitous in the central nervous system of vertebrates and invertebrates; at the same time, they also exist in the peripheral nervous muscle parts of invertebrates, making them excellent targets for insecticides, and have good insecticidal activity against pests such as ticks, fleas, lice, hemipterans and dipterans. Currently, four isoxazoline insecticides have been marketed: Fluralaner (fluralaner), Sarolaner (sarolaner), Afoxolaner (afoxolaner) and Lotilaner (lotilaner), which all contain a classic isoxazoline ring in their structure, and are mainly used for external insect control in animals such as cats and dogs.

[0004] Due to the excellent insecticidal activity and high safety of isoxazoline compounds, research on their derivatives has increased in recent years. In addition to the four isoxazoline drugs in the following table, other isoxazoline drugs are only used in the field of agriculture, such as Fluxametamide and Isocycloseram as broad-spectrum insecticides and acaricides for fruit trees and vegetables, cereals, rice, corn, soybeans, sugar beets, cotton and the like. Therefore, it is necessary to develop their use in veterinary medicine, i.e. safe administration to effectively control parasites in animals.

[0005] According to relevant patents and literature reports, the LD50 value of fluralaner and lotilaner is high, which proves that the safety of fluralaner and lotilaner is high. However, the higher the safety of a drug is, the larger the dose of the drug needs to be used. Fluralaner and lotilaner need a large amount to maintain an effective insecticidal concentration in the skin and blood, as shown in the following table:

[0006] Name LD50 (Rat) Dosage (Dog) Fluralaner > 2000 mg / kg 25 mg / kg Sarolaner 783 mg / kg 2 mg / kg Afoxolaner > 1000 mg / kg 2.5 mg / kg Lotilaner > 2000 mg / kg 20 mg / kg

[0007] In summary, there is an urgent need in the art to develop an isoxazoline antiparasitic drug with high insecticidal activity and good safety. SUMMARY

[0008] Based on the above, the present application aims to solve the technical problems in the prior art, and provides a derivative of an isoxazoline compound, a preparation method and application thereof. The technical scheme of the present application is as follows:

[0009] In a first aspect of the present application, a derivative of an isoxazoline compound is provided, and the structural general formula of the derivative of the isoxazoline compound is shown as formula (I):

[0010]

[0011] wherein R1 is selected from H, OH, CN, NO2, OCH3, CF3, N(CH3)2, Br or F;

[0012] R2 is selected from

[0013] Preferably, the derivative of the isoxazoline compound is selected from the following compounds 1-23:

[0014]

[0015]

[0016] The preparation method of the derivative of the isoxazoline compound is as follows:

[0017] Method 1: Preparation of compounds 1-10

[0018]

[0019] The reaction initiator A, chloro reagent (oxalyl chloride or sulfurous chloride, 1.1-1.5 eq) is added to the reaction bottle, and the reaction is carried out in a solvent (selected from tetrahydrofuran, dichloromethane, toluene, 5-15V, V is the volume mass ratio). During the reaction, thin layer chromatography (TLC) is used to detect the progress of the reaction. After the reaction is complete, the acyl chloride intermediate is obtained by concentrating to no fraction. Add ammonia water, after the reaction is complete, add 1M hydrochloric acid to terminate the reaction, add a solvent (selected from tetrahydrofuran, dichloromethane, ethyl acetate, 5-15V) to extract the aqueous phase, and the organic phase is concentrated and recrystallized to obtain the intermediate A.

[0020] The intermediate A, orthoformate reagent (selected from trimethyl orthoformate, triethyl orthoformate, triisopropyl orthoformate, 10.0-15.0 eq), nucleophilic reagent (selected from methylamine hydrochloride, S-methyl thiohydroxylamine hydrochloride, O-cyclopropyl hydroxylamine hydrochloride, methoxyamine hydrochloride, 1.0-1.5 eq) is added to the reaction bottle to carry out the reaction. During the reaction, thin layer chromatography (TLC) is used to detect the progress of the reaction. After the reaction is complete, the organic phase is filtered, concentrated, and recrystallized to obtain compound 1-10.

[0021] Method 2: Preparation of compounds 11-12

[0022]

[0023] The intermediate B, N,N-dimethylformamide (5.0-10.0V), N-bromosuccinimide (1.0-1.5 eq) and purified water (1.0-2.0V) are added to the reaction bottle and stirred to carry out the reaction. During the reaction, thin layer chromatography (TLC) is used to detect the progress of the reaction. After the reaction is complete, dilute with water, separate, and wash with water. The organic phase is concentrated and recrystallized to obtain compounds 11-12.

[0024] Method 3: Preparation of compound 13

[0025]

[0026] The reaction initiator 10, oxalyl chloride (1.1-1.5 eq) is added to the reaction bottle, and the reaction is carried out in a solvent (selected from tetrahydrofuran, dichloromethane, toluene, 5.0-15.0V). During the reaction, thin layer chromatography (TLC) is used to detect the progress of the reaction. After the reaction is complete, the acyl chloride intermediate is obtained by concentrating to no fraction. Add ammonia water, after the reaction is complete, add 1M hydrochloric acid to terminate the reaction, and the organic phase is concentrated and recrystallized to obtain compound 13.

[0027] Method 4: Preparation of compounds 14-23

[0028]

[0029] The reaction initiator B, oxalyl chloride (1.1-1.5 eq) is added to the reaction bottle, and the reaction is carried out in a solvent (selected from tetrahydrofuran, dichloromethane, toluene, 5.0-15.0V), and the reaction progress is monitored by thin layer chromatography (TLC) during the reaction, and after the reaction is completed, the acyl chloride intermediate is obtained by concentrating to no fraction.

[0030] The reaction initiator C (1.2-1.5 eq), triethylamine (3.0-5.0 eq) and a solvent (selected from tetrahydrofuran, dichloromethane, ethyl acetate, 5.0-15.0V) are added to the reaction bottle, and the reaction progress is monitored by thin layer chromatography (TLC) during the reaction, and after the reaction is completed, the reaction is terminated by adding 1M hydrochloric acid, and the organic phase is concentrated and recrystallized to obtain compounds 14-23.

[0031] In a second aspect of the present application, the use of the isoxazoline compound derivative in the preparation of a drug for preventing and treating parasitic infections is provided.

[0032] Preferably, the parasitic animal is an ectoparasite; the ectoparasite is selected from fleas, ticks, Demodex, scabies, ear mites, lice, mosquitoes, flies.

[0033] Preferably, the parasitic animal is an ectoparasite; the ectoparasite is selected from fleas, ticks, Demodex, scabies, ear mites, lice, mosquitoes, flies.

[0034] In a third aspect of the present application, the use of the isoxazoline compound derivative in the preparation of a drug for preventing and treating agricultural pests is provided.

[0035] Preferably, the agricultural pest is selected from Plutella xylostella, Mythimna separata, Spodoptera littoralis, Helicoverpa armigera, Ostrinia nubilalis, mosquito larvae, aphids, Tetranychus cinnabarinus.

[0036] In a fourth aspect of the present application, a pharmaceutical composition for preventing and treating parasitic infections is provided, which contains the isoxazoline compound derivative of the present application, and can further contain one or more pharmaceutically acceptable carriers and / or excipients.

[0037] In a fifth aspect of the present application, a pharmaceutical preparation for preventing and treating parasitic infections is provided, which contains the above-mentioned pharmaceutical composition.

[0038] Preferably, the pharmaceutical preparation is an external pharmaceutical preparation, including but not limited to drops, pouring agents, sprays, smearing agents, external solution, lotions, liniments, ointments, plasters, pastes, patches.

[0039] Preferably, the pharmaceutical preparation is an oral pharmaceutical preparation, including but not limited to tablets, granules, capsules, solutions, suspensions, emulsions.

[0040] Preferably, the pharmaceutical preparation is an injectable pharmaceutical preparation, including but not limited to injections, lyophilized powder.

[0041] Advantages:

[0042] (1) Compared with fluralaner and lotilaner, the 23 isoxazoline compound derivatives of the present application can achieve effective insecticidal effect at a lower dosage.

[0043] (2) Compared with fluralaner and lotilaner, the isoxazoline compound derivatives of the present application are safer.

[0044] (3) The isoxazoline compound derivatives of the present application have significant effects on combating animal ectoparasites, external environment and site pests, and agricultural pests. DETAILED DESCRIPTION

[0045] The above will be further described in detail through the specific embodiments in the form of examples. However, this should not be understood as limiting the scope of the above subject matter to the following examples. Any technology realized based on the content of the present application is within the scope.

[0046] The instruments, reagents, materials, etc. involved in the following examples, if not specifically stated, are conventional instruments, reagents, materials, etc. already existing in the prior art, which can be obtained through regular commercial channels. The experimental methods, detection methods, etc. involved in the following examples, if not specifically stated, are conventional experimental methods, detection methods, etc. already existing in the prior art.

[0047] Preparation method of compound 1 in example 1

[0048]

[0049] The starting material 1, oxalyl chloride (1.5 eq) (eq in the following examples means molar ratio) was added to the reaction bottle, and the reaction was carried out at 20-30°C in the solvent dichloromethane (10.0V, volume-mass ratio is 10ml:1g, V in the following examples is volume-mass ratio) for 12 hours. The reaction progress was detected by thin layer chromatography (TLC) during the reaction. After the reaction was completed, the acyl chloride intermediate was obtained by concentrating to no fraction, then ammonia water was added, and after the reaction was completed, 1M hydrochloric acid was added to terminate the reaction. The organic phase was extracted with dichloromethane (10.0V), and the organic phase was concentrated and recrystallized to obtain the intermediate 1.

[0050] The intermediate 1, triethyl orthoformate (15.0 eq), methylamine hydrochloride (1.5 eq) were added to the reaction bottle and the reaction was carried out at 50-60°C. The reaction progress was detected by thin layer chromatography (TLC) during the reaction. After the reaction was completed, the organic phase was filtered, concentrated and recrystallized to obtain compound 1.

[0051] Process for the preparation of compound 2 of example 2

[0052]

[0053] To the reaction flask was added starting material 1, oxalyl chloride (1.5 eq) and the reaction was carried out in dichloromethane (10.0 V) at 20-30 °C for 12 hours. The progress of the reaction was monitored by thin layer chromatography (TLC) during the reaction. After the reaction was complete, the acyl chloride intermediate was obtained by concentrating to dryness. Ammonia was added to the reaction flask and the reaction was allowed to complete. The reaction was quenched by adding 1 M hydrochloric acid. The aqueous phase was extracted with dichloromethane (10.0 V). The organic phase was concentrated and recrystallized to obtain intermediate 1.

[0054] To the reaction flask was added intermediate 1, trimethyl orthoformate (15.0 eq), S-methyl thiohydroxylamine hydrochloride (1.2 eq) and the reaction was carried out at 30-40 °C. The progress of the reaction was monitored by thin layer chromatography (TLC) during the reaction. After the reaction was complete, the organic phase was filtered, concentrated, and recrystallized to obtain compound 2.

[0055] Process for the preparation of compound 3 of example 3

[0056]

[0057] To the reaction flask was added starting material 1, oxalyl chloride (1.5 eq) and the reaction was carried out in dichloromethane (10.0 V) at 20-30 °C for 12 hours. The progress of the reaction was monitored by thin layer chromatography (TLC) during the reaction. After the reaction was complete, the acyl chloride intermediate was obtained by concentrating to dryness. Ammonia was added to the reaction flask and the reaction was allowed to complete. The reaction was quenched by adding 1 M hydrochloric acid. The aqueous phase was extracted with dichloromethane (10.0 V). The organic phase was concentrated and recrystallized to obtain intermediate 1.

[0058] To the reaction flask was added intermediate 1, triethyl orthoformate (13.0 eq), O-cyclopropylhydroxylamine hydrochloride (1.5 eq) and the reaction was carried out at 50-60 °C. The progress of the reaction was monitored by thin layer chromatography (TLC) during the reaction. After the reaction was complete, the organic phase was filtered, concentrated, and recrystallized to obtain compound 3.

[0059] Process for the preparation of compound 4 of example 4

[0060]

[0061] To the reaction flask was added starting material 4, thionyl chloride (1.5 eq) and the reaction was carried out in toluene (15.0 V) at 60-70 °C for 8 hours. The progress of the reaction was monitored by thin layer chromatography (TLC) during the reaction. After the reaction was complete, the acyl chloride intermediate was obtained by concentrating to dryness. Ammonia was added to the reaction flask and the reaction was allowed to complete. The reaction was quenched by adding 1 M hydrochloric acid. The aqueous phase was extracted with ethyl acetate (10.0 V). The organic phase was concentrated and recrystallized to obtain intermediate 4.

[0062] The intermediate 4, trimethyl orthoformate (13.0 eq), methoxyamine hydrochloride (1.3 eq) were added to the reaction bottle at 30-40 °C for reaction, the reaction progress was detected by thin layer chromatography (TLC) during the reaction, and after the reaction was completed, the organic phase was filtered, concentrated, and recrystallized to obtain compound 4.

[0063] Preparation method of compound 5 of example 5

[0064]

[0065] The starting material 5, thionyl chloride (1.5 eq) was added to the reaction bottle, and the reaction was carried out in toluene (10.0V) at 60-70 °C for 8 hours, the reaction progress was detected by thin layer chromatography (TLC) during the reaction, and after the reaction was completed, the acyl chloride intermediate was obtained by concentrating to no fraction, then adding ammonia water, after the reaction was completed, 1M hydrochloric acid was added to terminate the reaction, and the organic phase was extracted with ethyl acetate (10.0V), and the organic phase was concentrated and recrystallized to obtain intermediate 5.

[0066] The intermediate 5, triethyl orthoformate (14.0 eq), methoxyamine hydrochloride (1.3 eq) were added to the reaction bottle at 50-60 °C for reaction, the reaction progress was detected by thin layer chromatography (TLC) during the reaction, and after the reaction was completed, the organic phase was filtered, concentrated, and recrystallized to obtain compound 5.

[0067] Preparation method of compound 6 of example 6

[0068]

[0069] The starting material 6, oxalyl chloride (1.3 eq) was added to the reaction bottle, and the reaction was carried out in tetrahydrofuran (5.0V) at 20-30 °C for 8 hours, the reaction progress was detected by thin layer chromatography (TLC) during the reaction, and after the reaction was completed, the acyl chloride intermediate was obtained by concentrating to no fraction, then adding ammonia water, after the reaction was completed, 1M hydrochloric acid was added to terminate the reaction, and the organic phase was extracted with tetrahydrofuran (10.0V), and the organic phase was concentrated and recrystallized to obtain intermediate 6.

[0070] The intermediate 6, trimethyl orthoformate (10.0 eq), methoxyamine hydrochloride (1.1 eq) were added to the reaction bottle at 30-40 °C for reaction, the reaction progress was detected by thin layer chromatography (TLC) during the reaction, and after the reaction was completed, the organic phase was filtered, concentrated, and recrystallized to obtain compound 6.

[0071] Preparation method of compound 7 of example 7

[0072]

[0073] Into a reaction flask was placed starting material 7, thionyl chloride (1.5 eq) and the reaction was carried out in toluene (15.0 V) at 60-70 °C for 8 hours. The progress of the reaction was monitored by thin layer chromatography (TLC) during the reaction. After the reaction was completed, the acyl chloride intermediate was obtained by concentrating to dryness. Ammonia was added to the intermediate, and after the reaction was completed, the reaction was terminated by adding 1 M hydrochloric acid. The aqueous phase was extracted with ethyl acetate (10.0 V), and the organic phase was concentrated and recrystallized to obtain intermediate 7.

[0074] Into a reaction flask was placed intermediate 7, triisopropyl orthoformate (13.0 eq), and methoxyamine hydrochloride (1.2 eq), and the reaction was carried out at 60-70 °C. The progress of the reaction was monitored by thin layer chromatography (TLC) during the reaction. After the reaction was completed, the organic phase was filtered, concentrated, and recrystallized to obtain compound 7.

[0075] Method for preparing compound 8 of Example 8

[0076]

[0077] Into a reaction flask was placed starting material 8, oxalyl chloride (1.5 eq) and the reaction was carried out in tetrahydrofuran (5.0 V) at 20-30 °C for 8 hours. The progress of the reaction was monitored by thin layer chromatography (TLC) during the reaction. After the reaction was completed, the acyl chloride intermediate was obtained by concentrating to dryness. Ammonia was added to the intermediate, and after the reaction was completed, the reaction was terminated by adding 1 M hydrochloric acid. The aqueous phase was extracted with tetrahydrofuran (15.0 V), and the organic phase was concentrated and recrystallized to obtain intermediate 8.

[0078] Into a reaction flask was placed intermediate 8, triethyl orthoformate (15.0 eq), and methoxyamine hydrochloride (1.5 eq), and the reaction was carried out at 50-60 °C. The progress of the reaction was monitored by thin layer chromatography (TLC) during the reaction. After the reaction was completed, the organic phase was filtered, concentrated, and recrystallized to obtain compound 8.

[0079] Method for preparing compound 9 of Example 9

[0080]

[0081] Into a reaction flask was placed starting material 9, oxalyl chloride (1.3 eq) and the reaction was carried out in dichloromethane (10.0 V) at 20-30 °C for 12 hours. The progress of the reaction was monitored by thin layer chromatography (TLC) during the reaction. After the reaction was completed, the acyl chloride intermediate was obtained by concentrating to dryness. Ammonia was added to the intermediate, and after the reaction was completed, the reaction was terminated by adding 1 M hydrochloric acid. The aqueous phase was extracted with dichloromethane (10.0 V), and the organic phase was concentrated and recrystallized to obtain intermediate 9.

[0082] The intermediate 9, triethyl orthoformate (12.0 eq), methoxyamine hydrochloride (1.1 eq) were added to the reaction bottle at 50-60 °C for reaction, the reaction progress was detected by thin layer chromatography (TLC) during the reaction, and the organic phase was filtered, concentrated and recrystallized to obtain compound 9 after the reaction was completed.

[0083] Example 10 Preparation method of compound 10

[0084]

[0085] The starting material 10, oxalyl chloride (1.1 eq) was added to the reaction bottle, and the reaction was carried out in dichloromethane (5.0V) at 0-10 °C, the reaction progress was detected by thin layer chromatography (TLC) during the reaction, and the acyl chloride intermediate reaction solution was added to ammonia water after the reaction was completed, the reaction was terminated after 1M hydrochloric acid was added, and the organic phase was extracted with dichloromethane (5.0V), and the organic phase was concentrated and recrystallized to obtain intermediate 10.

[0086] The intermediate 10, triisopropyl orthoformate (15.0 eq), methoxyamine hydrochloride (1.5 eq) were added to the reaction bottle at 60-70 °C for reaction, the reaction progress was detected by thin layer chromatography (TLC) during the reaction, and the organic phase was filtered, concentrated and recrystallized to obtain compound 10 after the reaction was completed.

[0087] Example 11 Preparation method of compound 11

[0088]

[0089] The intermediate 1 described in Example 1, N,N-dimethylformamide (10.0V), N-bromosuccinimide (1.5 eq) and purified water (2.0V) were added to the reaction bottle at 60-70 °C for reaction, the reaction progress was detected by thin layer chromatography (TLC) during the reaction, and the reaction was diluted with water after the reaction was completed, the water phase was extracted with ethyl acetate (10.0V), and the organic phase was concentrated and recrystallized to obtain compound 11.

[0090] Example 12 Preparation method of compound 12

[0091]

[0092] The intermediate 10 described in Example 10, N,N-dimethylformamide (8.0V), N-bromosuccinimide (1.3 eq) and purified water (1.0V) were added to the reaction bottle at 60-70 °C for reaction, the reaction progress was detected by thin layer chromatography (TLC) during the reaction, and the reaction was diluted with water after the reaction was completed, the water phase was extracted with ethyl acetate (10.0V), and the organic phase was concentrated and recrystallized to obtain compound 12.

[0093] Method for preparing compound 13 of example 13

[0094]

[0095] The starting material 10, oxalyl chloride (1.1 eq) was added to the reaction bottle, and the reaction was carried out in dichloromethane (10.0V) at 0-10°C. The reaction progress was monitored by thin layer chromatography (TLC) during the reaction. After the reaction was completed, the acyl chloride intermediate reaction solution was added to ammonia water. After the reaction was completed, 1M hydrochloric acid was added to terminate the reaction. The water phase was extracted by dichloromethane (10.0V). The organic phase was concentrated and recrystallized to obtain compound 13 (intermediate 10).

[0096] Method for preparing compound 14 of example 14

[0097]

[0098] The starting material 1, oxalyl chloride (1.5 eq) was added to the reaction bottle, and the reaction was carried out in dichloromethane (10.0V) at 20-30°C for 12 hours. The reaction progress was monitored by thin layer chromatography (TLC) during the reaction. After the reaction was completed, it was concentrated to no fraction to obtain the acyl chloride intermediate. Methoxyamine was added, and after the reaction was completed, 1M hydrochloric acid was added to terminate the reaction. The water phase was extracted by dichloromethane (10.0V). The organic phase was concentrated and recrystallized to obtain compound 14.

[0099] Method for preparing compound 15 of example 15

[0100]

[0101] The starting material 15-1, oxalyl chloride (2.0 eq) was added to the reaction bottle, and the reaction was carried out in dichloromethane (15.0V) at 20-30°C for 12 hours. The reaction progress was monitored by thin layer chromatography (TLC) during the reaction. After the reaction was completed, it was concentrated to no fraction to obtain the acyl chloride intermediate.

[0102] The starting material 15-2 (1.3 eq), triethylamine (4.0 eq), and ethyl acetate (15.0V) were added to the reaction bottle and stirred until uniform. The acyl chloride intermediate was added to the reaction bottle. The reaction progress was monitored by thin layer chromatography (TLC) during the reaction. After the reaction was completed, 1M hydrochloric acid was added to terminate the reaction. The organic phase was concentrated and recrystallized to obtain compound 15.

[0103] Method for preparing compound 16 of example 16

[0104]

[0105] Into a reaction flask was placed starting material 15-1, oxalyl chloride (2.0 eq), and dichloromethane (15.0 V). The reaction was stirred at 20-30 °C for 12 hours. The reaction progress was monitored by thin layer chromatography (TLC). After the reaction was completed, the reaction mixture was concentrated to dryness to obtain the acyl chloride intermediate.

[0106] Into a reaction flask was placed starting material 16-2 (1.3 eq), triethylamine (5.0 eq), and tetrahydrofuran (5.0 V). The reaction was stirred until the starting material was dissolved. The acyl chloride intermediate was added to the reaction flask. The reaction progress was monitored by thin layer chromatography (TLC). After the reaction was completed, 1 M hydrochloric acid was added to quench the reaction. The organic phase was concentrated and recrystallized to obtain compound 16.

[0107] Example 17 Method of making compound 17

[0108]

[0109] Into a reaction flask was placed starting material 15-1, oxalyl chloride (2.0 eq), and dichloromethane (15.0 V). The reaction was stirred at 20-30 °C for 12 hours. The reaction progress was monitored by thin layer chromatography (TLC). After the reaction was completed, the reaction mixture was concentrated to dryness to obtain the acyl chloride intermediate.

[0110] Into a reaction flask was placed starting material 17-2 (1.3 eq), triethylamine (3.0 eq), and tetrahydrofuran (10.0 V). The reaction was stirred until the starting material was dissolved. The acyl chloride intermediate was added to the reaction flask. The reaction progress was monitored by thin layer chromatography (TLC). After the reaction was completed, 1 M hydrochloric acid was added to quench the reaction. The organic phase was concentrated and recrystallized to obtain compound 17.

[0111] Example 18 Method of making compound 18

[0112]

[0113] Into a reaction flask was placed starting material 15-1, oxalyl chloride (2.0 eq), and dichloromethane (15.0 V). The reaction was stirred at 20-30 °C for 12 hours. The reaction progress was monitored by thin layer chromatography (TLC). After the reaction was completed, the reaction mixture was concentrated to dryness to obtain the acyl chloride intermediate.

[0114] Into a reaction flask was placed starting material 18-2 (1.5 eq), triethylamine (5.0 eq), and tetrahydrofuran (5.0 V). The reaction was stirred until the starting material was dissolved. The acyl chloride intermediate was added to the reaction flask. The reaction progress was monitored by thin layer chromatography (TLC). After the reaction was completed, 1 M hydrochloric acid was added to quench the reaction. The organic phase was concentrated and recrystallized to obtain compound 18.

[0115] Example 19 Method of making compound 19

[0116]

[0117] Into a reaction flask was placed starting material 15-1, oxalyl chloride (2.0 eq) in dichloromethane (15.0 V) at 20-30 °C for 12 hours, the reaction progress was monitored by thin layer chromatography (TLC), after the reaction was completed, concentrated to no fraction to get acyl chloride intermediate.

[0118] Into a reaction flask was placed starting material 19-2 (1.2 eq), triethylamine (5.0 eq), tetrahydrofuran (5.0 V) and stirred to mix well, acyl chloride intermediate was added to the reaction flask, the reaction progress was monitored by thin layer chromatography (TLC), after the reaction was completed, 1M hydrochloric acid was added to terminate the reaction, the organic phase was concentrated and recrystallized to obtain compound 19.

[0119] Preparation method of compound 20 of example 20

[0120]

[0121] Into a reaction flask was placed starting material 4, sulfurous dichloride (1.5 eq) in toluene (10.0 V) at 60-70 °C for 8 hours, the reaction progress was monitored by thin layer chromatography (TLC), after the reaction was completed, concentrated to no fraction to get acyl chloride intermediate.

[0122] Into a reaction flask was placed starting material 20-2 (1.2 eq), triethylamine (5.0 eq), dichloromethane (15.0 V) and stirred to mix well, acyl chloride intermediate was added to the reaction flask, the reaction progress was monitored by thin layer chromatography (TLC), after the reaction was completed, 1M hydrochloric acid was added to terminate the reaction, the organic phase was concentrated and recrystallized to obtain compound 20.

[0123] Preparation method of compound 21 of example 21

[0124]

[0125] Into a reaction flask was placed starting material 7, sulfurous dichloride (1.5 eq) in toluene (15.0 V) at 60-70 °C for 8 hours, the reaction progress was monitored by thin layer chromatography (TLC), after the reaction was completed, concentrated to no fraction to get acyl chloride intermediate.

[0126] Into a reaction flask was placed starting material 20-2 (1.2 eq), triethylamine (4.0 eq), tetrahydrofuran (5.0 V) and stirred to mix well, acyl chloride intermediate was added to the reaction flask, the reaction progress was monitored by thin layer chromatography (TLC), after the reaction was completed, 1M hydrochloric acid was added to terminate the reaction, the organic phase was concentrated and recrystallized to obtain compound 21.

[0127] Preparation method of compound 22 of example 22

[0128]

[0129] Into a reaction flask was added starting material 5, thionyl chloride (1.5 eq) and toluene (15.0 V) and the reaction was allowed to proceed at 60-70 °C for 8 hours. The reaction progress was monitored by thin layer chromatography (TLC) and upon completion of the reaction, the reaction mixture was concentrated to dryness to obtain the acyl chloride intermediate.

[0130] Into a reaction flask was added starting material 20-2 (1.3 eq), triethylamine (5.0 eq), and tetrahydrofuran (5.0 V) and the mixture was stirred until homogeneous. The acyl chloride intermediate was added to the reaction flask and the reaction progress was monitored by thin layer chromatography (TLC). Upon completion of the reaction, 1 M hydrochloric acid was added to quench the reaction and the organic phase was concentrated and recrystallized to obtain compound 22.

[0131] Example 23 Method of preparing compound 23

[0132]

[0133] Into a reaction flask was added starting material 8, oxalyl chloride (1.5 eq) and tetrahydrofuran (15.0 V) and the reaction was allowed to proceed at 20-30 °C for 8 hours. The reaction progress was monitored by thin layer chromatography (TLC) and upon completion of the reaction, the reaction mixture was concentrated to dryness to obtain the acyl chloride intermediate.

[0134] Into a reaction flask was added starting material 20-2 (1.2 eq), triethylamine (3.0 eq), and tetrahydrofuran (5.0 V) and the mixture was stirred until homogeneous. The acyl chloride intermediate was added to the reaction flask and the reaction progress was monitored by thin layer chromatography (TLC). Upon completion of the reaction, 1 M hydrochloric acid was added to quench the reaction and the organic phase was concentrated and recrystallized to obtain compound 23.

[0135] The nuclear magnetic resonance hydrogen spectrum data and mass spectrum data of compounds 1-23 are shown in the following table:

[0136] Table 1 Nuclear magnetic resonance hydrogen spectrum data and mass spectrum data of compounds 1-23

[0137]

[0138]

[0139]

[0140]

[0141] Example 24 In vitro insecticidal toxicity test of the isoxazoline drug against parasitic infections of the present application

[0142] In vitro ticks: Compounds 1-23 were dissolved in DMSO and aliquots were added to citrated bovine blood in a petri dish covered with a membrane. The petri dish was placed on a warming tray. The fed ticks were removed and placed in a petri dish containing sand. Tick paralysis and / or death was observed at approximately 24, 48, and 72 hours. End point data was recorded as LC90 in μg / mL. The positive comparator was fluralaner and DMSO was used as the negative comparator.

[0143] In vitro mites: Compounds 1-23 were dissolved in DMSO and aliquots were added to citrated bovine blood in a petri dish covered with a membrane that was pre-warmed to 37°C. The fed mites were placed on the petri dish. Mortality of the fed mites was observed at approximately 24, 48, and 72 hours. End point data was recorded as LC90 in μg / mL. The positive comparator was fluralaner and DMSO was used as the negative comparator.

[0144] In vitro mosquitoes: Compounds 1-23 were dissolved in DMSO and aliquots were added to citrated bovine blood in a petri dish covered with a membrane. Approximately 10 mosquitoes were placed on each petri dish and covered. The mosquitoes were then allowed to feed on the treated bovine blood. The mosquitoes were maintained at approximately 80°F and a minimum of approximately 50% relative humidity. Knockdown and mortality of the mosquitoes was checked at approximately 2 and 24 hours. End point data was recorded as LC90 in μg / mL. The positive comparator was fluralaner and DMSO was used as the negative comparator.

[0145] In vitro fleas: Compounds 1-23 were dissolved in DMSO and aliquots were added to citrated bovine blood in a petri dish covered with a membrane that was pre-warmed to 37°C. The fed fleas were placed in a container on one side of an artificial membrane and the treated bovine blood was added to the container on the other side of the artificial membrane. Mortality of the fed fleas was observed at approximately 48, 96, and 128 hours. End point data was recorded as LC90 in μg / mL. The positive comparator was fluralaner and DMSO was used as the negative comparator.

[0146] Table 2 In vitro toxicity of isoxazoline compound derivatives - drug experiments

[0147]

[0148]

[0149] The results show that the isoxazoline compound derivatives of Examples 1-23, which were obtained by chemical synthesis, have a lower in vitro lethal concentration for mosquitoes, ticks, mites, and fleas compared to the marketed isoxazoline drug fluralaner, demonstrating a higher in vitro insecticidal activity.

[0150] Example 25 Oral toxicity experiment

[0151] Acute toxicity: the mice were observed and adaptively fed, the number of animals was determined according to the number of groups, the number of test animals (half of male and half of female) was selected for formal test, and 20 animals (half of male and half of female) were selected for each group.

[0152] Table 3 Isoxazoline derivative mouse acute toxicity experiment

[0153]

[0154]

[0155] Subchronic toxicity: the rats were observed and adaptively fed, the number of test animals (half of male and half of female) was selected for formal test, and 20 animals (half of male and half of female) were selected for each group, and 25mg of mixed beverage was given per day according to the dose of fluorofenirane, and the administration was continuously given for 30 days.

[0156] Table 4 Isoxazoline derivative experiment on the influence of body weight, food intake and water intake of rats

[0157]

[0158]

[0159] Table 5 Isoxazoline derivative experiment on the influence of blood routine index

[0160]

[0161] Table 6 Isoxazoline derivative experiment on the influence of organ coefficient of rats

[0162]

[0163]

[0164] The results show that the derivatives of the compounds prepared in examples 1-23 have higher safety and higher LD50; at the same time, no adverse reactions occur in the medication group in the subchronic toxicity test, the blood routine index and the organ coefficient have no obvious change compared with the non-medication group, which proves that the safety in the veterinary field is higher.

[0165] Example 26 Anti-parasite external preparation 1-23

[0166] Active ingredient: 500mg of compound 1-23 respectively

[0167] Solvent: 9ml of diethylene glycol monoethyl ether

[0168] Co-solvent: 1ml of dimethyl sulfoxide

[0169] Preparation method: take the prescribed amount of active ingredient, disperse in 80% diethylene glycol monoethyl ether, control the temperature at 30-40℃, add 1ml dimethyl sulfoxide, after clarification, add the remaining 20% diethylene glycol monoethyl ether, fill into low molecular weight polyethylene sealed tank, and it is obtained.

[0170] Example 27 Experiment on the effect of expelling ticks in vitro

[0171] According to the in vitro experimental data and the literature reports, the LD50 of ticks among the in vitro parasites is the highest, and the expelling and killing difficulty is the greatest, so ticks are selected for in vitro experiments on target animals.

[0172] In this evaluation, mixed gender beagles were used and assigned to a blank control group, a commercially available positive group (an external preparation of the active ingredient fluralaner) and Example 26 of the present application (external preparations 1-23 of the active ingredient compounds 1-23), and the experimental dogs were invaded by 50 un-fed adult ticks (Haemaphysalis longicornis).

[0173] At a dose of 40mg / kg of fluralaner and 10mg / kg of compounds 1-23, the dogs were treated on day 0. The preparation was administered using a pipette. The dose was applied in a line on the dorsal neck at the base of the skull.

[0174] The average detection rate of each group of drugs on the in vitro ticks of dogs was counted on day 0, 5, 10, 20 and 30 after administration, and on day 1, 2 and 7 after treatment, any immediate reaction of the dogs to the treatment was observed, and adverse reactions, skin irritation and properties of the test preparation after treatment were observed.

[0175] Table 7 Average detection rate of drugs on in vitro ticks

[0176]

[0177] The above experimental data show that the external preparations 1-23 prepared using the compounds 1-23 of the present application have better in vitro expelling effect than the commercially available products.

[0178] Example 28 Experiment on the effect of killing agricultural pests

[0179] (1) Plutella xylostella

[0180] To evaluate the control of the compounds of the present application on Plutella xylostella, the test unit is composed of a small open container with 12-14 day old radish plants inside. It is pre-infested with about 50 newly hatched larvae distributed in the test unit via corn cob grits. After being distributed into the test unit, the larvae move on the test plants.

[0181] Test compounds were prepared using a formulation comprising 20% dimethyl sulfoxide and 80% water. All test compounds in this test were sprayed at a concentration of 50 ppm, and the test was replicated three times in parallel. Test units were kept in a growth chamber at 25°C and 70% relative humidity for 6 days. Then, the plant feeding damage was visually evaluated according to the leaves eaten, and the mortality rate of the pests was also counted and calculated for each test unit.

[0182] Table 8 Average detection rate of the test compounds against Plutella xylostella

[0183] 0 days 6 days Blank 100％ 100％ Compound 1 100％ 1％ Compound 2 100％ 9％ Compound 3 100％ 6％ Compound 4 100％ 8％ Compound 5 100％ 10％ Compound 6 100％ 5％ Compound 7 100％ 14％ Compound 8 100％ 10％ Compound 9 100％ 17％ Compound 10 100％ 4％ Compound 11 100％ 3％ Compound 12 100％ 14％ Compound 13 100％ 9％ Compound 14 100％ 13％ Compound 15 100％ 5％ Compound 16 100％ 6％ Compound 17 100％ 8％ Compound 18 100％ 11％ Compound 19 100％ 4％ Compound 20 100％ 3％ Compound 21 100％ 20％ Compound 22 100％ 17％ Compound 23 100％ 13％

[0184] (2) Pseudaletia separata

[0185] To evaluate the control of the present compounds against Pseudaletia separata, test units were composed of a small open container with 4-5 day-old corn plants inside. They were pre-infested with 10 to 15 one-day-old larvae on a piece of insect food. Test compounds were prepared as described in Test (1) and sprayed at a concentration of 50 ppm, and the test was replicated three times in parallel. After spraying, the test units were left in a growth chamber, and then the control efficacy was evaluated for each test unit as described in Test (1).

[0186] Table 9 Average detection rate of the test compounds against Pseudaletia separata

[0187]

[0188]

[0189] (3) Aphis glycines

[0190] To evaluate the control effect of the present compounds against Aphis glycines, test units were composed of a small open container with 12 to 15 day-old radish plants inside. The plants were pre-infested by placing 30 to 40 aphids on a piece of leaf cut from the cultivated plants (cut leaf method) on a piece of leaf of the test plants.

[0191] Test compounds were prepared and sprayed as described in Test (1). All test compounds in these tests were sprayed at a concentration of 100 ppm, and the test was replicated three times in parallel. Test units were kept in a growth chamber at 19-21°C and 50-70% relative humidity for 6 days. Then, the mortality rate of the insects was visually evaluated for each test unit.

[0192] Table 10 Average detection rate of the test compounds against Aphis glycines

[0193]

[0194]

[0195] The above experiments show that compounds 1-23 of the present invention have good effects in controlling agricultural pests.

[0196] The above description is merely a specific 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. A derivative of an isoxazoline compound, characterized in that, The derivative is one of the following compounds: 。 2. Use of the derivative of the isoxazoline compound according to claim 1 in the preparation of a medicament for preventing and treating parasitic infection.

3. Use of the derivative of the isoxazoline compound according to claim 1 in the preparation of a medicament for preventing and treating agricultural pests.

4. Use according to claim 2, characterized in that, The parasitic is an ectoparasite of animals.

5. Use according to claim 4, characterized in that, The ectoparasite is one or more of fleas, ticks, Demodex, Sarcoptes, Otodectes, lice, mosquitoes, flies.

6. Use according to claim 2, characterized in that, The parasitic is a parasite in the place or environment where the animals live.

7. Use according to claim 3, characterized in that, The agricultural pest is one or more of Plutella xylostella, Mythimna separata, Spodoptera exigua, Helicoverpa armigera, Ostrinia nubilalis, mosquito larvae, aphids, Tetranychus cinnabarinus.

8. A pharmaceutical composition for preventing and treating parasitic infection, comprising the derivative of the isoxazoline compound according to claim 1, and one or more pharmaceutically acceptable carriers and / or excipients.

9. Use of the derivative of the compound according to claim 1 or the pharmaceutical composition according to claim 8 in the preparation of a pharmaceutical preparation for preventing and treating parasitic infection.

Images