Pyrazole derivative as well as preparation method and application thereof

Abstract

The invention relates to a pyrazole derivative as well as a preparation method and application thereof, and belongs to the technical field of medicines. Specifically, the invention discloses a pyrazole derivative as shown in a general formula (I), a pharmaceutical composition containing the pyrazole derivative and application of the pyrazole derivative in preparation of anti-parasitic drugs. The structural general formula (I) of the pyrazole derivative is # imgabs0. The pyrazole derivative disclosed by the invention has a remarkable effect in the aspects of resisting parasites, agricultural and forestry pests or environmental pests and the like, and has a wide application prospect.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and specifically relates to a pyrazole derivative and a preparation method and application thereof. Background Art

[0002] Cryptosporidium belongs to a genus of the order Phylum Coccidiales of the parasitic protozoa, which includes multiple species that infect humans and animals. Among them, "Cryptosporidium parvum" is the main cause of diarrhea in humans and economic animals. Cryptosporidium is a gastrointestinal parasite that is mainly transmitted through the fecal-oral route. The host is infected after ingesting contaminated food or water. Cryptosporidiosis is generally not obvious in people and animals with normal immune function, but in hosts with weak immune function, it causes severe diarrhea and even death.

[0003] So far, there is no specific drug for Cryptosporidium. Nitazoxanide, as a highly effective and broad-spectrum drug against parasites, bacteria and fungal infections in animals and humans, is the first and only drug approved by the U.S. Food and Drug Administration (FDA) for the treatment of Cryptosporidium. However, it is only used to treat children or adults with normal immune function and is ineffective for immunodeficient patients. Therefore, the development of new and effective anti-Cryptosporidium drugs is of great practical significance.

[0004] Pyrazole is a five-membered heterocyclic compound with two nitrogen atoms at positions 1 and 2 of the ring. It exists in three forms and is widely present as a core structure in many compounds with important agrochemical and pharmaceutical activities. It has low toxicity, high efficiency, and diverse structural transformations. Pyrazole can generate a large number of lead compounds after fusion with other cyclic groups. It is reported that pyrazole derivatives have antimicrobial, antitumor, antiviral, antituberculosis, antimalarial, antidiabetic, antidiuretic, anticonvulsant, antidepressant, antiobesity and antipyretic effects. However, the therapeutic effect of pyrazole derivatives on Cryptosporidium has not yet been clarified.

[0005] Therefore, it is necessary to explore and develop new pyrazole derivatives for use as anti-Cryptosporidium drugs. Summary of the invention

[0006] The present invention aims to solve the technical problems in the prior art, design and screen a series of novel pyrazole derivatives through high-throughput screening, and the derivatives have a wide range of anti-Cryptosporidium activity in the screening of new anti-Cryptosporidium drugs, laying a solid foundation for the development of new and highly effective anti-Cryptosporidium drugs.

[0007] To achieve the above purpose, the specific technical solution of the present invention is as follows:

[0008] In the first aspect of the present invention, there is provided a pyrazole derivative represented by the general formula (I), the structure of which is as follows:

[0009]

[0010] Wherein, X1 and X2 are each independently selected from nitrogen, oxygen, and carbon;

[0011] R1 is selected from F, Cl, OCH3, CH3, COOCH3, a benzene ring substituted by R5, a 5-7 membered aromatic heterocycle or condensed ring substituted by R5, a C3-C6 aliphatic ring or aliphatic heterocycle substituted by R5;

[0012] R5 is selected from one or any combination of F, Cl, OCH3, NH2, COONH2, COOH, COOCH3, CN, CF3, C1-C5 alkane or cycloalkane;

[0013] R2 is selected from H, Cl, F, CH3 or OCH3;

[0014] R3 and R4 are each independently selected from H, CN, CF3;

[0015] W is selected from CH2, R6 substituted or unsubstituted benzene ring, pyridine, pyrimidine, pyrazole, imidazole or W is absent;

[0016] R6 is selected from H, Cl, F, CH3, NH2 or CN;

[0017] Z is selected from R7 substituted or unsubstituted benzene ring, 5-7 membered aromatic heterocycle;

[0018] R7 is selected from H, Cl, F, CH3, OCH3, NH2, COONH2, CN or CF3.

[0019] In certain preferred embodiments, R1 is selected from a benzene ring substituted with R5, pyridine, pyrazole, imidazole, indole, quinoline, benzocycloheptane, cyclopropane, cyclobutane, cyclopentane or cyclohexane.

[0020] In certain preferred embodiments, Z is selected from a R7-substituted benzene ring, pyridine, indole, quinoline, benzocyclohexane or benzocycloheptane.

[0021] Preferably, the pyrazole derivative includes but is not limited to one of structures 1-35:

[0022]

[0023]

[0024]

[0025] The compounds of the general formula (I) mentioned above can also exist in the form of their salts, which are converted into compounds of the general formula I in vivo. For example, within the scope of the present invention, the compounds of the present invention are converted into pharmaceutically acceptable salts according to processes known in the art, and are used in the form of salts.

[0026] All tautomeric forms of the compounds of the general formula (I) of the present invention are included within the scope of the present invention. The compounds of the present invention may exist in specific geometric or stereoisomer forms. Additional asymmetric carbon atoms may exist in substituents such as alkyl, and all these isomers and their mixtures are included within the scope of the present invention.

[0027] In some preferred embodiments, the pharmaceutically acceptable salts include but are not limited to salts formed by the compound of formula (I) and the following acids: hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, acetic acid, trifluoroacetic acid, pyruvic acid, citric acid, tartaric acid, lactic acid, maleic acid, benzenesulfonic acid, succinic acid, methanesulfonic acid, p-toluenesulfonic acid, fumaric acid, salicylic acid or phenylacetic acid.

[0028] In a second aspect, the present invention provides a method for synthesizing a pyrazole derivative represented by general formula (I), the steps of which are as follows:

[0029]

[0030] Add solvent I, base, and starting material 2 to starting material 1 in sequence, raise the temperature to reflux, cool to room temperature, filter, wash the filter cake twice with water, dry the filter cake, recrystallize with solvent II, and dry to obtain an off-white solid, i.e., a pyrazole derivative (compound 1-35).

[0031] The solvent I is one or more of acetonitrile, toluene, methanol, ethanol, ethyl acetate, N,N-dimethylformamide, acetone, and 1,4-dioxane.

[0032] The base is one or more of triethylamine, N,N-diisopropylethylamine, pyridine, potassium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide, and lithium hydroxide.

[0033] The solvent II is one or more of methanol, ethanol and ethyl acetate.

[0034] The compound of the general formula (I) of the present invention can be obtained by the above or similar preparation methods, and the corresponding starting materials can be selected according to the different substituents and the different positions of the substituents. Those skilled in the art should recognize that the above route is helpful for understanding the present invention, but is not limited to the content of the present invention. Unless otherwise specified, the variables are defined as mentioned in the general formula (I).

[0035] In a third aspect, the present invention provides a pharmaceutical composition comprising the pyrazole derivative represented by the general formula (I) or its enantiomer, diastereomer, racemate, pharmaceutically acceptable salt or a mixture thereof and one or more pharmaceutically acceptable carriers and / or excipients.

[0036] The pharmaceutical composition of the present invention can be administered in various known ways, such as orally, parenterally, by spray or via an implanted reservoir. The pharmaceutical composition of the present invention can be administered alone or in combination with other drugs. Oral compositions can be in any orally acceptable dosage form, including but not limited to tablets, capsules, emulsions, suspensions, dispersions and solutions. Commonly used pharmaceutically acceptable carriers or excipients include stabilizers, diluents, surfactants, lubricants, antioxidants, adhesives, colorants, fillers, emulsifiers, etc.

[0037] The sterile injection composition can be formulated according to the techniques known in the art using suitable dispersants or wetting agents and suspending agents. Pharmaceutically acceptable carriers and solvents that can be used include water, mannitol, sodium chloride solution, and the like.

[0038] In a fourth aspect, the present invention provides the use of the pyrazole derivatives represented by the general formula (I) or the pharmaceutical composition in the preparation of drugs for treating parasitic infections or preventing and controlling agricultural pests or pests in living environments.

[0039] Preferably, the parasite is Cryptosporidium.

[0040] The agricultural pests are one or more of diamondback moth, armyworm, fall armyworm, cotton bollworm, corn borer, mosquito larvae, aphids, spider mites, whiteflies, psyllids, leafhoppers, scale insects and beetles.

[0041] The pests in the living environment are mosquitoes, flies and the like.

[0042] Beneficial effects:

[0043] The present invention reports for the first time a compound with a pyrazole structure having antiparasitic activity. The novel compound with a pyrazole structure has a significant effect on antiprotozoa, especially on Cryptosporidium, and lays a solid foundation for the development of new drugs. DETAILED DESCRIPTION

[0044] The preparation methods of the compounds of the general formula (I) of the present invention are described below in conjunction with specific examples, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be easily performed by those skilled in the art to which the present invention belongs.

[0045] The starting materials, reaction reagents, etc. used in the specific embodiments of the present invention are all commercially available. The present invention can adopt the salt-forming method commonly used in the art to prepare the salt form.

[0046] The experimental methods in the examples of the present invention where specific conditions are not specified are generally carried out under conventional conditions or under conditions recommended by raw material or product manufacturers.

[0047] The structures of all compounds in the examples were determined by waters QDa mass spectrometry and H NMR spectra ( 1 H-NMR) characterization.

[0048] Example 1

[0049] Preparation of compound 1

[0050]

[0051] 2.1 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 1.9 g of the starting reactant 2. The temperature was raised to reflux for 3 hours, cooled to room temperature, filtered, the filter cake was washed twice with water, dried, and recrystallized with methanol to obtain an off-white solid (i.e., compound 1). [M+H] + :381.2, 1 H-NMR(DMSO-d6,300MHz)δ:11.95(s,1H),7.92(s,1H),7.64-7.74(d,2H),7 .42-7.78(m,6H),7.38-7.40(m,2H),6.31(s,2H),2.12(s,3H),2.38(s,3H).

[0052] Example 2

[0053] Preparation of compound 2

[0054]

[0055] 2.2 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.3 g of N,N-diisopropylethylamine, and 1.9 g of the starting reactant 2. The reaction was heated to reflux for 4 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized with methanol to obtain an off-white solid (compound 2). [M+H] + :411.1, 1 H-NMR (DMSO-d6, 300MHz) δ: 13.95 (s, 1H), 8.24 (s, 1H), 7.95 (s, 1H), 7.64-7.75 (m, 3H), 7.45-7.63 (m, 3H), 7.38-7.40 (m, 2H), 2.42 (s, 3H).

[0056] Example 3

[0057] Preparation of compound 3

[0058]

[0059] 2.3 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.2 g of the starting reactant 2. The reaction was heated to reflux for 3.5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized with ethanol to obtain an off-white solid (compound 3). [M+H] + :409.2, 1 H-NMR(DMSO-d6,300MHz)δ:12.67(s,1H),7.85(s,1H),7.65-7.79(m,4H),7.45-7.72(m,4H),7.15-7.21(d,1H),2.46(s,3H),2.39(s,3H).

[0060] Example 4

[0061] Preparation of compound 4

[0062]

[0063] 2.4 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.2 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 3.5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized with ethanol to obtain an off-white solid (compound 4). [M+H] + :443.1, 1 H-NMR(DMSO-d6,300MHz)δ:12.65(s,1H),7.85(s,1H),7.65-7.79(m,2H),7 .45-7.62(m,4H),7.15-7.30(d,2H),7.11(s,1H),2.43(s,3H),2.47(s,3H).

[0064] Example 5

[0065] Preparation of compound 5

[0066]

[0067] 2.5 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.6 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 3.5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 5). [M+H] + :456.2, 1H-NMR(DMSO-d6,300MHz)δ:12.65(s,1H),7.87(s,1H),7.35-7.51(m,2H),7 .15-7.27(m,4H),6.91-7.05(m,3H),6.21(s,2H),3.82(s,3H),2.80(s,3H).

[0068] Example 6

[0069] Preparation of compound 6

[0070]

[0071] 2.5 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.5 g of the starting reactant 2. The reaction was heated to reflux for 4 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 6). [M+H] + :442.2, 1 H-NMR(DMSO-d6,300MHz)δ:13.45(s,1H),8.23(s,1H),7.87-8.02(d,2H),7.30 -7.48(d,2H),7.25-3.30(d,2H),6.91-7.05(d,2H),6.27(s,2H),3.82(s,3H).

[0072] Example 7

[0073] Preparation of compound 7

[0074]

[0075] 2.5 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 3.6 g of the starting reactant 2. The reaction was heated to reflux for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol:ethyl acetate (V:V=1:2) to obtain an off-white solid (Compound 7). [M+H] + :456.2, 1 H-NMR(DMSO-d6,300MHz)δ:12.55(s,1H),7.94(s,1H),7.48-7.60(m,3H),7.24-7. 32(d,2H),7.25(d,1H),6.89-7.05(d,2H),6.23(s,2H),3.86(s,3H),4.04(s,3H).

[0076] Example 8

[0077] Preparation of compound 8

[0078]

[0079] 2.4 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 3.4 g of the starting reactant 2. The reaction was heated to reflux for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol:ethyl acetate (V:V=1:3) to obtain an off-white solid (Compound 8). [M+H] + :484.2, 1 H-NMR(DMSO-d6,300MHz)δ:12.57(s,1H),8.60(s,1H),8.02-8.13(d,2H),7.65(m, 1H),7.41-7.52(m,2H),7.11-7.20(m,2H),6.24(s,2H),3.86(s,3H),4.05(s,3H).

[0080] Example 9

[0081] Preparation of compound 9

[0082]

[0083] 2.5 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.7 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 9). [M+H] + :463.2, 1 H-NMR(DMSO-d6,300MHz)δ:12.57(s,1H),8.15(s,1H),8.02-8.13(m,2H),7.85(s,1H),7 .72(m,1H),7.21-7.33(m,2H),6.85-6.97(m,2H),6.27(s,2H),3.87(s,3H),4.06(s,3H).

[0084] Example 10

[0085] Preparation of compound 10

[0086]

[0087] 2.6 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.7 g of the starting reactant 2. The reaction was heated to reflux for 4 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 10). [M+H] + :510.2, 1 H-NMR(DMSO-d6,300MHz)δ:12.57(s,1H),8.10(s,1H),7.94(s,1H),7.62-7.78(m, 2H),7.45-7.54(m,2H),7.40(m,1H),7.22-7.35(m,2H),6.27(s,2H),4.06(s,3H).

[0088] Embodiment 11

[0089] Preparation of compound 11

[0090]

[0091] 2.6 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.4 g of the starting reactant 2. The reaction was heated to reflux for 4 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 11). [M+H] + :469.2, 1 H-NMR(DMSO-d6,300MHz)δ:12.57(s,1H),12.20(s,1H),8.62(s,1H),8.12-8.25(m,2H),7.9 4(s,1H),7.71-7.80(m,1H),7.43-7.55(m,2H),7.22-7.35(m,2H),2.41(s,3H),2.45(s,3H).

[0092] Example 12

[0093] Preparation of compound 12

[0094]

[0095] 2.6 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 3.0 g of the starting reactant 2. The reaction was heated to reflux for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 12). [M+H] + :443.2, 1H-NMR(DMSO-d6,300MHz)δ:12.57(s,1H),7.94(s,1H),7.42-7.60(m,3H),7.44-7.51(d,1H),7.02-7.22(m,4H),5.32(s,2H),2.45(s,3H).

[0096] Embodiment 13

[0097] Preparation of compound 13

[0098]

[0099] 3.0 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.0 g of the starting reactant 2. The reaction was heated to reflux for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethyl acetate to obtain an off-white solid (Compound 13). [M+H] + :450.2, 1 H-NMR(DMSO-d6,300MHz)δ:11.84(s,1H),7.47-7.56(m,1H),7.31-7.44(m,3H),7.28(s, 1H),6.41-6.53(m,1H),3.20(m,1H),2.47(s,3H),1.48-1.53(m,1H),1.13-1.25(m,1H).

[0100] Embodiment 14

[0101] Preparation of compound 14

[0102]

[0103] 2.7 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.2 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethyl acetate to obtain an off-white solid (Compound 14). [M+H] + :439.2, 1 H-NMR(DMSO-d6,300MHz)δ:11.80(s,1H),7.94(s,1H),7.76(s,2H),7.31-7.48(m,2H),7.18-7.28(m,2H),3.75 -3.85(m,1H),3.11-3.23(m,1H),2.34-2.45(m,1H),3.20(m,1H),2.47(s,3H),2.13-2.21(m,1H),1.82(s,3H).

[0104] Embodiment 15

[0105] Preparation of compound 15

[0106]

[0107] 2.2 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.5 g of the starting reactant 2. The reaction was heated to reflux for 4 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethyl acetate to obtain an off-white solid (Compound 15). [M+H] + :436.2, 1 H-NMR(DMSO-d6,300MHz)δ:11.80(s,1H),7.94(s,1H),7.82-7.95(m,2H),7.43-7.58(m,2H) ,6.71(s,1H),3.42(m,1H),2.90(s,3H),2.71(m,1H),1.65-1.86(m,4H),1.50-1.70(m,4H).

[0108] Example 16

[0109] Preparation of compound 16

[0110]

[0111] 1.3 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.4 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 6 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethyl acetate to obtain an off-white solid (Compound 16). [M+H] + :352.2, 1 H-NMR(DMSO-d6,300MHz)δ:12.55(s,1H),7.94(s,1H),7.41-7.53(m,2H),7 .29(s,1H),7.03-7.29(m,4H),6.43-6.65(m,2H),5.28(s,2H),3.81(s,3H).

[0112] Embodiment 17

[0113] Preparation of compound 17

[0114]

[0115] 1.8 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.4 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 4 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 17). [M+H] + :366.2, 1 H-NMR(DMSO-d6,300MHz)δ:12.55(s,1H),8.79-8.84(d,1H),8.31-8.39(d,1H),7.94(s,1H),7.90 (s,1H),7.71-7.82(m,2H),7.45-7.50(m,1H),6.67(s,1H),5.80(s,2H),2.48(s,3H),2.43(s,3H).

[0116] Embodiment 18

[0117] Preparation of compound 18

[0118]

[0119] 1.7 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.4 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 18). [M+H] + :358.2, 1 H-NMR(DMSO-d6,300MHz)δ:12.55(s,1H),7.94(s,1H),7.42-7.61(m,3H),7 .12-7.22(m,2H),7.69(s,1H),6.48-6.55(d,1H),6.41(s,1H),2.48(s,3H).

[0120] Embodiment 19

[0121] Preparation of compound 19

[0122]

[0123] 2.8 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.3 g of the starting reactant 2. The reaction was heated to reflux for 4 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 19). [M+H] + :464.2, 1H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),7.96(s,1H),7.94(s,1H),7.55-7.66(m,2H),7.25-7. 42(m,3H),6.87(s,1H),6.68-6.72(d,1H),6.25(s,2H),4.61(s,2H),2.48(s,3H),2.10(s,3H).

[0124] Embodiment 20

[0125] Preparation of compound 20

[0126]

[0127] 2.5 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.2 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 20). [M+H] + :439.2, 1 H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),8.74(s,1H),8.12(s,1H),7.80(s,1H),7.69-7.74(m, 1H),7.30-7.50(m,2H),6.90-7.19(m,2H),6.90(s,1H),3.87(s,3H),2.45(s,3H),2.14(s,3H).

[0128] Embodiment 21

[0129] Preparation of compound 21

[0130]

[0131] 2.1 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 3.2 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 4.5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 21). [M+H] + :450.3, 1H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),7.94(s,1H),7.48-7.60(m,5H),6.80(s,1H),6.68(s,1H ),6.56(s,2H),6.27(s,2H),2.50-2.63(m,4H),2.50-2.63(m,4H),2.45(s,3H),1.62-1.75(m,4H).

[0132] Embodiment 22

[0133] Preparation of compound 22

[0134]

[0135] 2.6 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 3.2 g of the starting reactant 2. The reaction was heated to reflux for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 22). [M+H] + :494.2, 1 H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),8.12(s,1H),7.48-7.60(m,5H),7.45(s,2H),7.41(s,1H),6.31(s,1H ),6.27(s,2H),6.21(s,1H),2.93-3.04(m,2H),2.63-2.73(m,2H),2.15(s,3H),2.11(s,3H),1.82-1.95(m,2H).

[0136] Embodiment 23

[0137] Preparation of compound 23

[0138]

[0139] 2.1 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 3.4 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 23). [M+H] + :465.3, 1H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),7.64(s,1H),7.48-7.60(m,5H),7.45(s,2H),6.31(s,1H),6.27(s,2H ),6.21(s,1H),5.04(s,1H),2.93-3.04(m,2H),2.63-2.73(m,2H),2.15(s,3H),2.11(s,3H),1.74-1.93(m,4H).

[0140] Embodiment 24

[0141] Preparation of compound 24

[0142]

[0143] 2.1 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.5 g of the starting reactant 2. The reaction was heated to reflux for 4 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 24). [M+H] + :434.2, 1 H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),8.34-8.39(d,1H),7.74-7.84(d,1H),7.45-7.62(m,5H),7. 32(s,1H),6.92-7.03(d,1H),6.31-6.42(d,1H),6.25(s,2H),3.74(s,3H),2.47(s,3H),2.12(s,3H).

[0144] Embodiment 25

[0145] Preparation of compound 25

[0146]

[0147] 2.1 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 1.9 g of the starting reactant 2. The reaction was heated to reflux for 6 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 25). [M+H] + :323.2, 1 H-NMR (DMSO-d6, 300MHz) δ: 11.85 (s, 1H), 7.94 (s, 1H), 7.45-7.62 (m, 5H), 6.28 (s, 2H), 2.47 (s, 3H), 2.12 (s, 3H).

[0148] Embodiment 26

[0149] Preparation of compound 26

[0150]

[0151] 2.3 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.2 g of the starting reactant 2. The reaction was heated to reflux for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 26). [M+H] + :361.1, 1 H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),9.29(s,1H),8.76(s,1H),7.94(s,1H),7.78-7.90(m ,2H),7.48-7.61(d,1H),7.23-7.35(m,1H),6.72-6.87(m,1H),6.42-6.54(d,1H),2.47(s,3H).

[0152] Embodiment 27

[0153] Preparation of compound 27

[0154]

[0155] 1.9 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.3 g of the starting reactant 2. The reaction was heated to reflux for 4.5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 27). [M+H] + :314.1, 1 H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),7.94(s,1H),7.76-7.87(d,1H),7.55 -7.68(d,1H),7.01-7.27(m,2H),6.27-6.39(d,1H),4.06(s,3H),2.47(s,3H).

[0156] Embodiment 28

[0157] Preparation of Compound 28

[0158]

[0159] 2.3 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 1.6 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 28). [M+H] + :339.1, 1 H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),7.94(s,1H),7.83(s,1H),7.16-7.34(m,3H),6.89(s,1H),2.44-2.51(m,6H),2.27(s,3H).

[0160] Embodiment 29

[0161] Preparation of compound 29

[0162]

[0163] 2.9 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 1.9 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 29). [M+H] + :442.1, 1 H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),8.09(s,1H),7.94(s,1H),7.62-7.72(d ,1H),7.46-7.58(m,2H),7.35-7.41(t,1H),7.13-7.24(d,1H),3.77-3.82(m,6H).

[0164] Embodiment 30

[0165] Preparation of compound 30

[0166]

[0167] 2.2 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.3 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 30). [M+H] + :407.2, 1H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),8.82(m,1H),8.21(s,1H),7.94(s,1H),7.63-7.79(m ,3H),7.46-7.58(m,3H),7.35-7.41(d,1H),7.13-7.24(m,2H),3.79(s,3H),2.43-2.52(m,6H).

[0168] Embodiment 31

[0169] Preparation of compound 31

[0170]

[0171] 2.7 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 1.8 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 6 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 31). [M+H] + :377.2, 1 H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),7.94(s,1H),6.73-6.91(m,3H),6.02 (s,2H),2.75(m,2H),2.70(m,2H),2.45(s,3H),2.10(s,3H),1.65-1.77(m,4H).

[0172] Embodiment 32

[0173] Preparation of compound 32

[0174]

[0175] 2.8 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 2.1 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 32). [M+H] + :390.2, 1 H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),7.94(s,1H),7.03-7.26(m,3H),2.73-2.86 (m,4H),2.75(s,3H),2.47(s,3H),2.12(s,3H),1.79-1.85(m,2H),1.63-1.76(m,4H).

[0176] Embodiment 33

[0177] Preparation of compound 33

[0178]

[0179] 2.3 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 1.8 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 33). [M+H] + :341.2, 1 H-NMR (DMSO-d6, 300MHz) δ: 11.85 (s, 1H), 7.94 (s, 1H), 8.38 (d, 2H), 6.88-6.99 (m, 2H), 2.75 (s, 3H), 2.47 (s, 3H), 2.12 (s, 3H).

[0180] Embodiment 34

[0181] Preparation of compound 34

[0182]

[0183] 2.5 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 1.6 g of the starting reactant 2. The reaction was heated to reflux for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 34). [M+H] + :361.2, 1 H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),10.84(s,1H),7.94(s,1H),7.85(d,1H),7 .64(s,1H),7.52(d,1H),6.84-7.03(m,2H),2.75(s,3H),2.47(s,3H),2.12(s,3H).

[0184] Embodiment 35

[0185] Preparation of compound 35

[0186]

[0187] 2.6 g of the starting reactant 1 was added to a three-necked flask, followed by 20 mL of acetonitrile, 3.8 g of N,N-diisopropylethylamine, and 1.8 g of the starting reactant 2. The reaction was refluxed at elevated temperature for 5 hours, cooled to room temperature, filtered, and the filter cake was washed twice with water. The filter cake was dried and recrystallized from ethanol to obtain an off-white solid (Compound 35). [M+H] + :373.2, 1 H-NMR(DMSO-d6,300MHz)δ:11.85(s,1H),8.91(d,1H),8.35(d,1H),7.94(s, 1H),7.85-7.97(m,3H),7.53(d,1H),2.75(s,3H),2.47(s,3H),2.12(s,3H).

[0188] Example 36 Human colon cancer (HCT-8) cells were tested for anti-Cryptosporidium in vitro activity

[0189] 1. Reagents: RPMI1640, trypsin, sodium deoxycholate, fetal bovine serum and other reagents were purchased from commercial sources.

[0190] 2. Experimental Cryptosporidium species: Bovine Cryptosporidium oocysts were collected from cow feces, sieved through 60 mesh, and stored in 2.5% potassium dichromate (K2Cr2O7) preservation solution at 4°C for use.

[0191] 3. Experimental Methods

[0192] 3.1 Oocyst purification

[0193] 10 mL of fecal suspension was drawn from 2.5% K2Cr2O7 preservation solution, washed twice with 3 times the volume of purified water, and then made into fecal suspension with 2 times the volume of purified water to purify the oocysts. 30 mL of saturated sucrose solution was added to a 50 mL centrifuge tube, 10 mL of fecal resuspension was slowly dripped on it, centrifuged at 1500 g for 20 minutes at 4°C, 5 mL of surface liquid was drawn, and 3 times the volume of purified water was added to wash twice, and centrifuged at 1500 g for 20 minutes, and set aside.

[0194] 3.2 Sterilization of oocysts

[0195] 2×10 purified Cryptosporidium andersoni oocysts were drawn from 2.5% K2Cr2O7 storage solution. 5 oocysts, washed twice with purified water to remove K2Cr2O7, resuspended to 900 μL in sterile purified water in a 2 mL EP tube, cooled on ice and sterilized. During sterilization, 100 μL of ice-cold 1.05% sodium hypochlorite solution was added to the oocyst suspension, shaken vigorously and placed on ice for 5 minutes, washed twice with sterile PBS buffer and resuspended to 500 μL.

[0196] 3.3 Excystation of Sterile Cryptosporidium Oocysts

[0197] Add 12 times the amount of oocyst excystation solution (50 mg pancreatin and 150 mg sodium taurocholate, dissolved in 10 mL PBS, sterilized with a 0.22 μm filter) to the sterile 500 μL oocyst suspension and place in a 37°C water bath for 40 minutes. After excystation, wash twice with sterile PBS buffer, resuspend to 1 mL with RPMI-1640 basal culture medium containing 10% FCS, and set aside.

[0198] 3.4 Cell culture

[0199] HCT-8 cells were cultured in RPMI-1640 medium containing 5% fetal calf serum (FCS) in a 37°C 5% CO2 cell culture incubator. After the cells covered the bottom of the culture dish, they were digested with cell digestion solution and transferred to new culture dishes.

[0200] 3.5 Infection of HCT-8 cells

[0201] HCT-8 cells were cultured in 6-well plates using RPMI-1640 medium containing 5% fetal bovine serum and placed in a 37°C 5% CO2 cell culture incubator for 48 hours. 3 Add the excysted oocysts and continue culturing for 8 hours, and incubate with complete medium for 48 hours.

[0202] 3.6 Anti-Cryptosporidium Activity of Test Compounds against HCT-8 Cells Infected with Cryptosporidium

[0203] Nitazoxanide was used as a control drug, and a positive control group and a blank control group were set up. The test compound was administered according to 6 concentration gradients (200 μg / mL, 40 μg / mL, 8 μg / mL, 1.6 μg / mL, 0.32 μg / mL, 0.064 μg / mL) and added to the infected HCT-8 cells. The cells were incubated for 48 hours to evaluate the pharmacodynamic activity.

[0204] Table 1 Results of in vitro anti-Cryptosporidium activity test of HTC-8 cells

[0205] Serial number <![CDATA[IC 50 (μM)]]> Serial number <![CDATA[IC 50 (μM)]]> Serial number <![CDATA[IC 50 (μM)]]> Nitazoxanide 0.11±0.021 Compound 12 0.09±0.056 Compound 24 0.052±0.019 Compound 1 0.082±0.015 Compound 13 0.12±0.09 Compound 25 0.054±0.031 Compound 2 0.079±0.020 Compound 14 0.14±0.10 Compound 26 0.084±0.031 Compound 3 0.052±0.032 Compound 15 0.15±0.11 Compound 27 0.075±0.015 Compound 4 0.053±0.018 Compound 16 0.15±0.09 Compound 28 0.09±0.023 Compound 5 0.12±0.09 Compound 17 0.06±0.023 Compound 29 0.087±0.019 Compound 6 0.10±0.089 Compound 18 0.05±0.012 Compound 30 0.076±0.024 Compound 7 0.13±0.033 Compound 19 0.12±0.086 Compound 31 0.081±0.038 Compound 8 0.12±0.023 Compound 20 0.18±0.11 Compound 32 0.054±0.011 Compound 9 0.13±0.068 Compound 21 0.089±0.03 Compound 33 0.043±0.014 Compound 10 0.11±0.072 Compound 22 0.067±0.028 Compound 34 0.076±0.021 Compound 11 0.10±0.021 Compound 23 0.037±0.02 Compound 35 0.062±0.026

[0206] Experimental conclusion: The in vitro anti-Cryptosporidium activity of the compounds in the examples of the present invention is better than that of nitazoxanide or equivalent to that of nitazoxanide, and has development value.

[0207] Example 37 In vivo anti-cryptosporidiosis activity test in animals

[0208] 1. Experimental animals: 3-week-old ICR mice were purchased from Shanghai JST Laboratory Animal Co., Ltd.

[0209] 2. Test methods:

[0210] 2.1 Grouping and inoculation of mice: The experimental mice were divided into groups, and nitazoxanide was used as a control drug. A positive control group and a blank control group were set up, and the sterilized Cryptosporidium oocysts prepared in Example 36 were inoculated.

[0211] 2.2 Dosing regimen for mice: Four days after inoculation, mice were gavaged with 200 μL per mouse. The dose of nitazoxanide was 100 mg / kg, and the dose of the test compound was 100 mg / kg and 200 mg / kg. The drugs were administered for 15 consecutive days. The blank control group was not treated.

[0212] 2.3 Collection of mouse fecal samples:

[0213] One day after inoculation, mouse feces were collected and collected continuously for 20 days. Two portions of feces were collected from each group every day, each portion weighing 1 g. The feces were transferred into 5 mL centrifuge tubes and a small amount of water was added. One portion was stored at 4°C for subsequent oocyst counting, and the other was stored at -20°C for future use.

[0214] 2.4 Cryptosporidium oocyst count:

[0215] Dissolve the mouse feces collected from each group every day in 40 mL of water, stir evenly with a tongue depressor, filter through four layers of gauze, take 5 mL of the filtrate, centrifuge at 3000 r / min for 10 minutes, and discard the supernatant. Add 10 mL of saturated sugar water to the precipitate, mix evenly, centrifuge at 1500 r / min for 10 minutes. Take 1 drop of the liquid surface and add it to a slide, cover it with a slide, and examine it under a 400x microscope. Use a counter to record the number of oocysts.

[0216] Calculate the oocyst reduction rate of each experimental group as follows: oocyst reduction rate (%) = (total number of oocysts expelled in the infection group - total number of oocysts expelled in the experimental group) ÷ total number of oocysts expelled in the infection group × 100%.

[0217] Table 2 Oocyst reduction rate in mice

[0218]

[0219]

[0220] Experimental conclusion: The anti-Cryptosporidium activity of the compounds in the examples of the present invention in vivo is better than that of nitazoxanide or equivalent to that of nitazoxanide.

[0221] Example 38 In vitro toxicity test on Plutella xylostella

[0222] The diamondback moth larvae were randomly divided into blank control, flurellana control drug group and experimental group. The concentration of the drug to be tested was set to 100ug / mL. Thirty diamondback moth larvae were placed on 12-day-old radish plants in each experimental group, and each group of radish plants was medicated by spraying. After 48 hours, the plant feeding damage was assessed based on the leaves ingested and the mortality of diamondback moth was counted. The diamondback moth toxicity results are shown in Table 3.

[0223] Table 3 Detection rate of drug toxicity to Plutella xylostella

[0224]

[0225]

[0226] The results show that the compounds of the present invention have activity in killing Plutella xylostella, and the activity of the compounds in the examples is better than that of flurella or is equivalent to that of flurella.

Claims

1. A pyrazole derivative represented by general formula (I): in, X1 and X2 are each independently selected from nitrogen, oxygen, and carbon; R1 is selected from F, Cl, OCH3, CH3, COOCH3, a benzene ring substituted by R5, a 5-7 membered aromatic heterocycle or condensed ring substituted by R5, a C3-C6 aliphatic ring or aliphatic heterocycle substituted by R5; R2 is selected from H, Cl, F, CH3 or OCH3; R3 and R4 are each independently selected from H, CN, CF3; W is selected from CH2, R6 substituted or unsubstituted benzene ring, pyridine, pyrimidine, pyrazole, imidazole or W is absent; Z is selected from a benzene ring or a 5-7 membered aromatic heterocyclic ring substituted or unsubstituted by R7.

2. The pyrazole derivative according to claim 1, characterized in that: The R5 is selected from one or any combination of F, Cl, OCH3, NH2, COONH2, COOH, COOCH3, CN, CF3, C1-C5 alkanes or cycloalkanes; the R6 is selected from H, Cl, F, CH3, NH2 or CN; the R7 is selected from H, Cl, F, CH3, OCH3, NH2, COONH2, CN or CF3.

3. The pyrazole derivative according to claim 1, characterized in that: R1 is selected from a benzene ring substituted with R5, pyridine, pyrazole, imidazole, indole, quinoline, benzocycloheptane, cyclopropane, cyclobutane, cyclopentane or cyclohexane.

4. The pyrazole derivative according to claim 1, characterized in that: Z is selected from a benzene ring substituted with R7, pyridine, indole, quinoline, benzocyclohexane or benzocycloheptane.

5. The pyrazole derivative according to claim 1, characterized in that: The derivative is one of structures 1-35:

6. The pyrazole derivative according to claim 1, characterized in that: The salts of the pyrazole derivatives represented by the general formula (I) include salts formed by the pyrazole derivatives represented by the general formula (I) and the following acids: hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, acetic acid, trifluoroacetic acid, pyruvic acid, citric acid, tartaric acid, lactic acid, maleic acid, benzenesulfonic acid, succinic acid, methanesulfonic acid, p-toluenesulfonic acid, fumaric acid, salicylic acid or phenylacetic acid.

7. A pharmaceutical composition comprising the pyrazole derivative represented by the general formula (I) according to any one of claims 1 to 6 and one or more pharmaceutically acceptable carriers and / or excipients.

8. Use of the pyrazole derivative represented by the general formula (I) as described in any one of claims 1 to 6 or the pharmaceutical composition as described in claim 7 in the preparation of a medicament for treating parasitic infections in humans and / or animals, agricultural pests or pest infestations in living environments.

9. The use according to claim 8, characterized in that The parasite is Cryptosporidium.

10. The use according to claim 8, characterized in that The agricultural pests are one or more of diamondback moth, armyworm, fall armyworm, cotton bollworm, corn borer, mosquito larvae, aphids, spider mites, whiteflies, psyllids, leafhoppers, scale insects and beetles.

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