Cyclocarbamide-containing thiazole compound as well as preparation method and application thereof

Abstract

The invention relates to the technical field of agricultural protection, in particular to a cyclamine urea-containing thiazole compound as well as a preparation method and application thereof. The cyclamine urea-containing thiazole compound provided by the invention is a compound as shown in a formula (1), the drug solubility is improved by prolonging a carbon chain structure among nitazoxanide molecules, the activity of the compound is further improved, drug molecules of an azacycloalkane structure have good insecticidal and bacteriostatic effects, the two active molecule fragments are spliced, and the compound has good insecticidal and bacteriostatic effects. The purposes of optimizing the medicine structure and improving the biological activity can be achieved. Formula (1): # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the technical field of agricultural protection, and in particular to a thiazole compound containing cyclamino urea, a preparation method thereof and an application thereof. Background Art

[0002] Cryptosporidiosis is a foodborne and waterborne infectious disease caused by Cryptosporidium parvum, which infects a variety of mammals (J Parasit Dis, 2022, 46(3), 923-939). More than 800 cases of human cryptosporidiosis have been reported annually (Int J Parasitol, 2018, 48(1), 1-12), causing a significant impact worldwide. Currently, more than 40 different Cryptosporidium species have been identified, of which Cryptosporidium hominis and Cryptosporidium parvum are the main causes of disease (Pak Vet J, 2023, 43(2), 213-223). The main host of Cryptosporidium is livestock, resulting in a decline in livestock production and economic downturn (EST, 2017, 51(15), 8663-8671).

[0003] Currently, nitazoxanide is still the only FDA-approved drug for the treatment of cryptosporidiosis in the world (Expert Rev Anti-infe, 2023, 21(2), 167-173). The therapeutic mechanism is still unclear. Studies have shown that its metabolite, nitazoxanide, may inhibit the growth of Cryptosporidium oocysts by blocking the reaction of pyruvate-ferredoxin oxidoreductase (PFOR enzyme) (IJBM, 2023, 247, 125823). Nitazoxanide is only effective for patients with normal immune function, and its efficacy will be greatly reduced for patients with immune deficiency and children (Expert Opin Pharmaco, 2021, 22(17), 2337-2342). Therefore, the development of new veterinary insecticides targeting Cryptosporidium has become an urgent need and research hotspot. In this context, we selected nitazoxanide as the research object to guide the design and synthesis of new drugs.

[0004] Since its development in the last century, nitazoxanide (NTZ) has been widely commercialized worldwide. As the first US Food and Drug Administration (FDA)-approved treatment for Cryptosporidium infection and currently the only commercially available treatment for the disease, NTZ has developed a certain degree of drug resistance after long-term use. Furthermore, due to incomplete research on NTZ, no complete structure-activity relationship has been reported for Cryptosporidium infection. In addition, no such products are currently marketed or imported in my country. Therefore, the development of new and highly effective drug molecules has become a research hotspot both domestically and internationally. Therefore, research on nitazoxanide derivatives holds great promise.

[0005] The inventors discovered that extending the carbon chain structure between nitazoxanide molecules has a positive effect on improving drug solubility and activity. Furthermore, the azacycloalkane structure of the drug molecule has good insecticidal and antibacterial effects. By splicing the two active molecular fragments, they hope to optimize the drug structure and enhance its biological activity. Summary of the Invention

[0006] The existing technology has problems such as drug resistance to nitazoxanide and unclear complete structure-activity relationship for Cryptosporidium. Provided is a thiazole compound containing cyclohexane urea, which has good biological activity and can be used for the prevention and treatment of cryptosporidiosis.

[0007] In order to achieve the above-mentioned object, the present invention provides a thiazole compound containing cyclamine urea, which is a compound represented by formula (1);

[0008] Formula (1):

[0009] Wherein, R1 is -R4-R5-O-R6, and R4 is selected from C3-C 15 Azacycloalkyl; R5 is selected from C1-C 10 Alkylene; R6 is selected from substituted or unsubstituted C6-C 20 aryl, or selected from substituted or unsubstituted C3-C 12 Heteroaryl; substituted C6-C 20 Aryl and substituted C3-C 12 The substituents of the heteroaryl group are selected from C1-C 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 At least one of a haloalkyl group and a halogen;

[0010] R2 is selected from H, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C3-C 20 At least one of the cycloalkyl groups; substituted C1-C 12 Alkyl, substituted C2-C 12 Alkenyl and substituted C3-C 20 The substituent of the cycloalkyl group is selected from at least one of a C1-C6 alkyl group, a C2-C6 alkenyl group, and a C3-C8 cycloalkyl group;

[0011] Each R3 is independently selected from H, C1-C 12 At least one of an alkyl group, a nitro group and a cyano group.

[0012] The second aspect of the present invention provides a method for preparing the thiazole compound containing cyclanilamide according to the first aspect of the present invention, the method comprising:

[0013] S1: The compound represented by formula (A) and the compound represented by formula (B) undergo a first contact reaction to obtain a compound represented by formula (C);

[0014] S2: subjecting the compound represented by formula (C) to a second contact reaction with the compound represented by formula (D) to prepare the thiazole compound containing cyclic amine urea;

[0015] Formula (A): Formula (B):

[0016] Formula (C): Formula (D): H-R1;

[0017] wherein each R7 is independently selected from H or nitro, and at least one is nitro; and X is halogen.

[0018] The third aspect of the present invention provides a use of the thiazole compound containing cyclanilamide according to the first aspect of the present invention in preventing and controlling Cryptosporidium.

[0019] The fourth aspect of the present invention provides a use of the thiazole compound containing cyclamino urea according to the first aspect of the present invention in preventing and controlling plant pathogens.

[0020] The aforementioned technical solution extends the carbon chain structure of nitazoxanide molecules, positively impacting their solubility and activity. Furthermore, the azacycloalkane-based drug molecule exhibits significant insecticidal and antibacterial effects. By splicing the two active molecular fragments, the drug's structure can be optimized to enhance its biological activity. DETAILED DESCRIPTION

[0021] The endpoints of the ranges and any values disclosed herein are not limited to the precise ranges or values, and these ranges or values should be understood to include values close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

[0022] In one aspect, the present invention provides a thiazole compound containing cycloamine urea, which is a compound represented by formula (1);

[0023] Formula (1):

[0024] Wherein, R1 is -R4-R5-O-R6, and R4 is selected from C3-C 15Azacycloalkyl; R5 is selected from C1-C 10 Alkylene; R6 is selected from substituted or unsubstituted C6-C 20 aryl, or selected from substituted or unsubstituted C3-C 12 Heteroaryl; substituted C6-C 20 Aryl and substituted C3-C 12 The substituents of the heteroaryl group are selected from C1-C 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 At least one of a haloalkyl group and a halogen;

[0025] R2 is selected from H, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C3-C 20 At least one of the cycloalkyl groups; substituted C1-C 12 Alkyl, substituted C2-C 12 Alkenyl and substituted C3-C 20 The substituent of the cycloalkyl group is selected from at least one of a C1-C6 alkyl group, a C2-C6 alkenyl group, and a C3-C8 cycloalkyl group;

[0026] Each R3 is independently selected from H, C1-C 12 At least one of an alkyl group, a nitro group and a cyano group.

[0027] In the present invention, in order to improve the activity of the thiazole compound containing cycloamine urea, preferably, in the compound represented by formula (1), R4 is selected from C3-C 10 R5 is selected from C1-C6 alkylene; R6 is selected from substituted or unsubstituted C6-C 14 aryl and substituted or unsubstituted C3-C 10 Heteroaryl; substituted C6-C 14 Aryl and substituted C3-C 10 The substituent of the heteroaryl is selected from at least one of a C1-C6 alkyl, a C1-C6 alkoxy, a C1-C6 haloalkyl and a halogen;

[0028] R2 is selected from H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl and substituted or unsubstituted C3-C 10 At least one of cycloalkyl; substituted C1-C6 alkyl, substituted C2-C6 alkenyl and substituted C3-C 10 The substituent of the cycloalkyl group is selected from at least one of a C1-C3 alkyl group, a C2-C4 alkenyl group, and a C3-C6 cycloalkyl group;

[0029] Each R3 is independently selected from at least one of H, a C1-C8 alkyl group, a nitro group, and a cyano group.

[0030] In the present invention, preferably, R4 is selected from at least one of azetidinyl, pyrrolidinyl, piperidinyl, azepanyl, azooctanyl, azanonyl, azadecyl and piperazinyl;

[0031] R5 is selected from at least one of -CH2-, -CH2-CH2-, -CH2-CH2-CH2-, -CH(CH3)-CH2-, -CH2-CH(CH3)-, -CH2-(CH2)2-CH2-, -C(CH3)2-CH2- and -CH2-C(CH3)2-;

[0032] R6 is selected from at least one of substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, substituted or unsubstituted pyridyl, substituted or unsubstituted piperidyl and substituted or unsubstituted quinolyl; the substituents of the substituted phenyl, substituted naphthyl, substituted pyridyl, substituted piperidyl and substituted quinolyl are selected from -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -CH2CH2CH2CH3, -CH2CH(CH3)2, -CH(CH3)CH2CH3, -C(CH3) 3, at least one of -OCH3, -OCH2CH3, -OCH2CH2CH3, -OCH(CH3)2, -OCH2CH2CH2CH3, -OCH2CH(CH3)2, -OCH(CH3)CH2CH3, -OC(CH3)3, -CH2F, -CH2Cl, -CH2Br, -CH2I, -CHF2, -CHCl2, -CHBr2, -CHI2, -CF3, -CCl3, -CBr3, -CI3, F, Cl, Br and I;

[0033] R2 is selected from at least one of H, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -CH2CH2CH2CH3, -CH2CH(CH3)2, -CH(CH3)CH2CH3, -C(CH3)3, -CH2CH2CH2CH2CH3, -CH2CH2CH(CH3)2, -CH2C(CH3)3, -CH(C H3)C H2CH2C H3, -C H2C H2C H2C H2C H3, -CH(CH3)CH2CH2CH2CH3, -CH2CH(CH3)CH2CH2CH3 and -C(CH3)2CH2CH2CH3;

[0034] Each R3 is independently selected from at least one of H, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -CH2CH2CH2CH3, -CH2CH(CH3)2, -CH(CH3)CH2CH3, -C(CH3)3 and nitro.

[0035] In the present invention, preferably, the compound is a compound represented by formula (2);

[0036] Formula (2):

[0037] wherein n=1-8, for example, n=1, n=2, n=3, n=4, n=5, n=6, n=7, n=8;

[0038] In the present invention, preferably, n=1-5, for example, n=1, n=2, n=3, n=4, and n=5.

[0039] In the present invention, the inventors have found that when the azacycloalkyl group in the compound is an azetidinyl group, the compound has excellent activity. Therefore, preferably, in the compound represented by formula (2), n=1.

[0040] In the present invention, preferably, the compound represented by formula (2) is selected from at least one of the following compounds:

[0041] Formula (d1): Formula (d2):

[0042] Formula (d3): Formula (d4):

[0043] Formula (d5): Formula (d6):

[0044] Formula (d7): Formula (d8):

[0045] Formula (d9): Formula (d10):

[0046] Formula (d11): Formula (d12):

[0047] Formula (d13): Formula (d14):

[0048] In the present invention, the thiazole compound containing cyclohexane urea also has excellent activity when containing a pyrrolidine group. According to a preferred embodiment, in the compound represented by formula (2), n=2;

[0049] In the present invention, preferably, the compound represented by formula (2) is selected from at least one of the following compounds:

[0050] Formula (h1): Formula (h2):

[0051] Formula (h3): Formula (h4):

[0052] According to another preferred embodiment, in order to prepare thiazole compounds containing cyclic semicarbazide with excellent activity, the nitrogen heterocycloalkyl group can also be a piperidinyl group. For this purpose, in the compound represented by formula (2), n=3.

[0053] In the present invention, preferably, the compound represented by formula (2) is selected from at least one of the following compounds:

[0054] Formula (j1): Formula (j2):

[0055] Formula (j3): Formula (j4):

[0056] The second aspect of the present invention provides a method for preparing the thiazole compound containing cyclanilamide according to the first aspect of the present invention, the method comprising:

[0057] S1: The compound represented by formula (A) and the compound represented by formula (B) undergo a first contact reaction to obtain a compound represented by formula (C);

[0058] S2: subjecting the compound represented by formula (C) to a second contact reaction with the compound represented by formula (D) to prepare the thiazole compound containing cyclic amine urea;

[0059] Formula (A): Formula (B):

[0060] Formula (C): Formula (D): H-R1;

[0061] wherein each R7 is independently selected from H or nitro, and at least one is nitro; and X is halogen.

[0062] In the present invention, the first contact reaction is to connect the amino group in the compound represented by formula (A) with the carbonate group in the compound represented by formula (B) to generate an amino group protected by carbonate, that is, to generate the compound represented by formula (C).

[0063] In the present invention, preferably, the conditions for the first contact reaction include: temperature of 0-30°C, time of 0.5-5h; more preferably, the conditions for the first contact reaction include: temperature of 0-10°C, time of 1-3h.

[0064] In the present invention, in the first contact reaction, the amino group in the compound represented by formula (A) needs to be deprotonated in an alkaline environment to generate an amino anion. To this end, preferably, the first contact reaction is carried out in the presence of a first alkaline reagent, wherein the first alkaline reagent is selected from at least one of triethylamine (Et3N), N,N-diisopropylethylamine (DIPEA), and pyridine. More preferably, the molar ratio of the compound represented by formula (A) to the first alkaline reagent is 1:1-4, preferably 1:1.5-3, for example, 1:1.5, 1:1.8, 1:2, 1:3, and any range therebetween.

[0065] In the present invention, the first contact reaction is carried out in the presence of a first solvent. Preferably, the first solvent is selected from at least one of tetrahydrofuran (THF), diethyl ether, and dichloromethane (DCM). More preferably, the mass ratio of the compound represented by formula (A) to the first solvent is 1:20-40, preferably 1:25-35, for example, 1:25, 1:30, 1:35, and any range therebetween.

[0066] In the present invention, the second contact reaction is the replacement of the carbonate protecting group on the compound represented by formula (C) with the R1 group in the compound represented by formula (D), thereby preparing the thiazole compound containing cyclic semicarbazide. As previously described, the R1 group includes an amino group located on a heterocyclic ring. Therefore, this reaction is essentially the reaction of the amino group on the heterocyclic ring in the compound represented by formula (D) with the carbonate carbonyl carbon in the compound represented by formula (C) to form an amide bond, thereby preparing the thiazole compound containing cyclic semicarbazide.

[0067] In the present invention, preferably, the conditions for the second contact reaction include: temperature of 20-40°C and time of 3-10 hours; more preferably, the conditions for the second contact reaction include: temperature of 25-30°C and time of 6-10 hours.

[0068] In the present invention, preferably, the second contact reaction is carried out in the presence of a second solvent, wherein the second solvent is selected from at least one of tetrahydrofuran (THF), diethyl ether, and dichloromethane (DCM). More preferably, the mass ratio of the compound represented by formula (C) to the second solvent is 1:20-40, preferably 1:25-35, for example, 1:25, 1:30, 1:35, and any range therebetween.

[0069] In the present invention, preferably, the compound represented by formula (D) can be obtained by sequentially subjecting the compound represented by formula (a) to sulfonylation reaction, substitution reaction and hydrolysis reaction;

[0070] Formula (a):

[0071] In the present invention, preferably, the sulfonylation reaction is carried out by combining the hydroxyl group in the compound represented by formula (a) with the sulfonyl group in the sulfonylation agent to prepare the compound represented by formula (b);

[0072] Formula (b):

[0073] Wherein, M is selected from at least one of Ts, Ms and Tf.

[0074] In the present invention, preferably, the conditions for the sulfonylation reaction include: temperature of 20-40° C., and time of 6-18 h; more preferably, the conditions for the sulfonylation reaction include: temperature of 25-35° C., and time of 12-16 h.

[0075] In the present invention, preferably, the sulfonylating agent is selected from at least one of TsCl, MsCl, and TfCl. More preferably, the molar ratio of the compound represented by formula (a) to the sulfonylating agent is 1:1-3, preferably 1:1.1-2, for example, 1:1.2, 1:1.5, 1:2, and any range therebetween.

[0076] In the present invention, the combination of the hydroxyl group in the compound represented by formula (a) and the sulfonyl group in the sulfonylating agent needs to be carried out in an alkaline environment. To promote the sulfonylation reaction, the sulfonylation reaction is preferably carried out in the presence of a second alkaline agent, wherein the second alkaline agent is selected from at least one of triethylamine (Et3N), N,N-diisopropylethylamine (DIPEA), and pyridine. More preferably, the molar ratio of the compound represented by formula (a) to the second alkaline agent is 1:1-5, preferably 1:1.5-3, for example, 1:1.5, 1:2, 1:3, and any range therebetween.

[0077] In the present invention, the sulfonylation reaction is preferably carried out in the presence of a third solvent, wherein the third solvent is selected from at least one of tetrahydrofuran (THF), diethyl ether, and dichloromethane (DCM). More preferably, the mass ratio of the compound represented by formula (a) to the third solvent is 1:15-40, preferably 1:18-35, for example, 1:18, 1:19, 1:25, 1:30, and any range therebetween.

[0078] In the present invention, preferably, the sulfonylation reaction further comprises purifying the reaction solution to prepare a compound represented by formula (b); the purification treatment comprises: after the sulfonylation reaction is completed, adding 1M NaOH solution for washing, DCM extraction, saturated brine washing, anhydrous sodium sulfate drying, and reduced pressure desolventizing to prepare the compound represented by formula (b).

[0079] In the present invention, preferably, the substitution reaction comprises: in the presence of a catalyst, performing a substitution reaction on the compound represented by formula (b) and the compound represented by formula (c) to prepare a compound represented by formula (d);

[0080] Formula (c): R6-PH; Formula (d):

[0081] In the present invention, the substitution reaction first converts the phenolic group in the compound represented by formula (c) into an oxygen anion in the presence of a catalyst, and then attacks the sulfonyl group in the compound represented by formula (b) to generate the compound represented by formula (d).

[0082] In the present invention, preferably, the conditions for the substitution reaction include: temperature of 50-100° C., and time of 6-15 h; more preferably, the conditions for the substitution reaction include: temperature of 60-90° C., and time of 8-12 h.

[0083] In the present invention, preferably, the catalyst is selected from at least one of CS2CO3, K2CO3, and Rb2CO3. More preferably, the molar ratio of the compound represented by formula (c) to the catalyst is 1:1-3, preferably 1:1.5-2.5, for example, 1:1.5, 1:2, 1:2.5, and any range therebetween.

[0084] In the present invention, the substitution reaction is preferably carried out in the presence of a fourth solvent, the fourth solvent being selected from at least one of dimethylformamide (DMF), N-methylpyrrolidone (NMP), and N,N-dimethylacetamide (DMAC). More preferably, the mass ratio of the compound represented by formula (b) to the fourth solvent is 1:3-20, preferably 1:5-15, for example, 1:5, 1:6, 1:7, 1:8, 1:10, 1:15, and any range therebetween.

[0085] In the present invention, preferably, the substitution reaction includes purifying the reaction solution to prepare the compound represented by formula (d); the purification treatment includes: after the substitution reaction is completed, adding water, DCM extraction, washing with saturated brine, adding anhydrous sodium sulfate for drying, and desolvating the solution under reduced pressure to prepare the compound represented by formula (d).

[0086] In the present invention, preferably, the hydrolysis reaction comprises: removing the amino protecting group in the compound of formula (d) using an acidic reagent to prepare the compound of formula (D).

[0087] In the present invention, preferably, the conditions for the hydrolysis reaction include: temperature of 0-20°C, and time of 3-8 hours; more preferably, the conditions for the hydrolysis reaction include: temperature of 0-10°C, and time of 4-6 hours.

[0088] In the present invention, preferably, the acidic agent is selected from at least one of trifluoroacetic acid (TFA), HCl, and H2SO4. More preferably, the molar ratio of the compound of formula (d) to the acidic agent is 1:10-45, preferably 1:25-40, for example, 1:25, 1:30, 1:35, 1:38, 1:40, and any range therebetween.

[0089] In the present invention, preferably, the hydrolysis reaction is carried out in the presence of a fifth solvent, wherein the fifth solvent is selected from at least one of tetrahydrofuran (THF), diethyl ether, and dichloromethane (DCM). More preferably, the mass ratio of the compound represented by formula (d) to the fifth solvent is 1:10-40, preferably 1:15-30, for example, 1:15, 1:18, 1:22, 1:26, 1:30, and any range therebetween.

[0090] In the present invention, preferably, the hydrolysis reaction includes purifying the reaction solution to prepare the compound represented by formula (D); the purification treatment includes: after the hydrolysis reaction is completed, vacuum desolventizing, adding ice water, adjusting the pH to 8-9, DCM extraction, washing with saturated brine, drying over anhydrous sodium sulfate, and vacuum desolventizing to prepare the compound represented by formula (D).

[0091] The preparation process of the thiazole compound containing cyclanilamide provided in the second aspect of the present invention is as follows:

[0092]

[0093] R1-R3 in the compound represented by the reaction product formula (1) are provided by the corresponding groups in the compound represented by formula (A), the compound represented by formula (B) and the compound represented by formula (D), and the reagents, reaction conditions and amounts of each raw material involved in the reaction formula can be appropriately selected from the reagents, reaction conditions and amounts of each raw material described in the aforementioned preparation method of thiazole compounds containing cyclic amine urea, and the present invention will not be repeated here.

[0094] In the present invention, when the compound represented by formula (A) is a compound represented by formula (A'), and the compound represented by formula (B) is a compound represented by formula (B'), the preparation process of the thiazole compound containing cyclic aminourea can be as follows:

[0095]

[0096] In the present invention, preferably, the preparation method of the thiazole compound containing cyclic aminourea further includes post-treatment of the reaction solution, and the post-treatment includes: filtering the reaction solution, desolventizing the filtrate, and purifying by column chromatography (the eluent is tetrahydrofuran: petroleum ether in a ratio of 1:3-8, for example, the eluent is tetrahydrofuran: petroleum ether = 1:5), to prepare the compound represented by formula (1) / the compound represented by formula (1').

[0097] The third aspect of the present invention provides a use of the thiazole compound containing cyclanilamide according to the first aspect of the present invention in preventing and controlling Cryptosporidium.

[0098] In the present invention, preferably, the Cryptosporidium is Cryptosporidium containing PFOR enzyme.

[0099] In the present invention, preferably, the Cryptosporidium is Cryptosporidium parvum.

[0100] The fourth aspect of the present invention provides a use of the thiazole compound containing cyclamino urea according to the first aspect of the present invention in preventing and controlling plant pathogens.

[0101] In the present invention, preferably, the plant pathogenic bacteria are plant pathogenic bacteria containing PFOR enzyme.

[0102] In the present invention, preferably, the plant pathogenic bacteria include at least one of Brassica rapa, Xanthomonas oryzae var. oryzae, R. solanacearum, Angular leaf spot of cucumber, Xanthomonas citri var. citri and Pseudomonas syringae pv. oryzae bacterial leaf streak.

[0103] The thiazole compounds containing cyclohexane urea prepared by the above technical scheme have excellent activity. According to some preferred embodiments, the thiazole compounds containing cyclohexane urea have a certain anti-Cryptosporidium effect, and the inhibition rate of some compounds is higher than 70%. At a concentration of 50 mg / L, most of the thiazole compounds containing cyclohexane urea have a certain inhibitory effect on bacteria such as rice stripe, rice bacterial blight, rice wilt, cabbage soft rot and citrus ulcer, and the fungicidal activity of some compounds can be higher than 90%.

[0104] The present invention will be described in detail below through examples.

[0105] Examples 1-22

[0106] This example is used to illustrate the preparation methods of the compounds represented by formulas d1-d14, j1-j4, and h1-h4. The specific preparation processes are as follows:

[0107]

[0108] The specific preparation method is as follows:

[0109] (1) At room temperature (25°C, the same below), 10 mmol of the compound represented by formula (a), 11 mmol of TsCl, and 20 mmol of triethylamine (Et3N) were placed in 30 mL of dichloromethane (DCM) solution and stirred for 15 h to perform a sulfonylation reaction. After the reaction was completed, the mixture was washed with 1 M NaOH solution, extracted three times with DCM, washed with saturated brine, dried over anhydrous sodium sulfate, and desolvated under reduced pressure to obtain the compound represented by formula (b);

[0110] (2) 10 mmol of the compound represented by formula (b), 10 mmol of the compound represented by formula (c) and 20 mmol of cesium carbonate (Cs2CO3) were placed in 20 mL of dimethylformamide (DMF) and subjected to substitution reaction at 80°C for 10 h. After the reaction was completed, water was added, DCM was extracted three times, saturated brine was washed, anhydrous sodium sulfate was added for drying, and the solution was decompressed to obtain the compound represented by formula (d);

[0111] (3) 10 mmol of the compound represented by formula (d) and 380 mmol of trifluoroacetic acid (TFA) were placed in 30 mL of dichloromethane (DCM) and subjected to a hydrolysis reaction at 0°C for 4 h. After the reaction was completed, the mixture was desolvated under reduced pressure, ice water was added, the pH was adjusted to 9, DCM was extracted three times, washed with saturated brine, dried over anhydrous sodium sulfate, and desolvated under reduced pressure to obtain the compound represented by formula (D);

[0112] (4) 4.6 mmol of 2-amino-5-nitrothiazole (compound represented by formula (A')), 4.6 mmol of nitrophenyl chloroformate (compound represented by formula (B')), and 8.28 mmol of N,N-diisopropylethylamine (DIPEA) were placed in 20 mL of tetrahydrofuran (THF) solution, and a first contact reaction was carried out at 0°C for 1.5 hours to prepare a compound represented by formula (C'), and then 4.6 mmol of the compound represented by formula (D) was added, and a second contact reaction was carried out at room temperature for 10 hours;

[0113] (5) After the reaction in step (4) is completed, the reaction solution is filtered, the filtrate is desolvated under reduced pressure, and purified by column chromatography (eluent is petroleum ether:tetrahydrofuran = 5:1) to prepare the compound represented by formula (1');

[0114] Among them, when the compound represented by formula (a) and the compound represented by formula (c) shown in Table 1 are used, the corresponding compounds represented by formula (1') prepared are compounds represented by formulas d1-d14, compounds represented by formulas j1-j4, and compounds represented by formulas h1-h4, and the yield of step (4) is shown in the following table;

[0115] Table 1

[0116]

[0117]

[0118]

[0119] Test Example 1

[0120] The anti-Cryptosporidium activity of the thiazole compounds containing cyclohexane prepared in the above examples was determined, and nitazoxanide was used as a control drug. The anti-Cryptosporidium activity test results of the thiazole compounds containing cyclohexane and nitazoxanide are shown in Table 2.

[0121] Test method:

[0122] 1. According to 2×10 4 cells and 10 4 The infection model was established with a dose of oocysts per well.

[0123] 2. 3 h after infection, the medium was changed to wash away the uninvaded parasites, and 200 μL of 2% FBS 1640 medium (final concentration of compound 10 μM, paromomycin 160 μM, DMSO 0.5%) was added to each well. The cells were cultured in a 37°C incubator for 41 h.

[0124] 3. Extract RNA and perform qPCR analysis of human and Cryptosporidium 18S. Calculate the inhibition rate based on the results of three replicates.

[0125] 4. The remaining compounds were tested against Cryptosporidium according to the above procedure.

[0126] Table 2

[0127] serial number Inhibition rate serial number Inhibition rate serial number Inhibition rate serial number Inhibition rate Formula d1 39 Formula d6 5 Formula d12 56 Formula j1 0 Formula d2 0 Formula d7 0 Formula d13 0 Formula j2 0 Formula d3 40 Formula d8 26 Formula d14 0 Formula j3 0 Formula d4 74 Formula d9 0 Formula h1 0 Formula d5 0 Formula d10 57 Formula h2 0 Nitazoxanide 42 Formula d11 0 Formula h3 0

[0128] It can be seen from Table 2 that most of the thiazole compounds containing cyclohexane urea prepared in Examples 1-22 have a certain anti-Cryptosporidium effect, and the inhibition rate of some compounds is higher than 70%.

[0129] Test Example 2

[0130] The anti-plant pathogenic fungal activity of the thiazole compounds containing cyclohexane prepared in the above examples was determined. Meanwhile, commercial drugs chloroquine, streptomycin sulfate, and 8-hydroxyquinoline copper were used as control groups. The anti-plant pathogenic fungal activity test results of the thiazole compounds containing cyclohexane and the control group are shown in Table 3.

[0131] Test method:

[0132] 1. Activation of bacterial strains: Take out the bacterial strains stored at -80℃ and streak them on the prepared NA medium plates. Then, invert and culture them in a constant temperature box at 28℃ in the dark until good, pure and single colonies grow.

[0133] 2. Prepare bacterial suspension: Transfer a single activated colony to sterilized NB liquid medium that has been cooled to room temperature. Incubate in the dark on a shaker at 28°C and 180 rpm. Adjust the incubation time based on the growth characteristics of the individual bacteria. Measure the OD value using a spectrophotometer and adjust the suspension to 1.0 using liquid NB medium. Shake thoroughly and set aside.

[0134] 3. Preparation of the drug stock solution: Weigh 5 mg of the drug to be tested using an analytical balance, add 1000 μL of DMSO to dissolve it, and use a vortex to mix to make a 5000 mg / L drug stock solution for use.

[0135] 4. Experimental Preparation: Spray a sterile 24-well plate with alcohol and place it in a clean bench. Turn on the UV sterilizer for at least 30 minutes. Sterilize the required pipette tips and liquid NB culture medium in an autoclave at 121°C for 25 minutes. After sterilization, spray the plate with alcohol and place it in a clean bench under UV sterilization for at least 30 minutes.

[0136] 5. Primary Screening Inhibition Rate Determination: Add a bacterial suspension with an OD value of 1.0 to liquid NB medium at a 1% inoculum volume. Shake well, then pipette 990 μL into a 24-well plate. After pipetting, transfer 10 μL of the test drug stock solution to three parallel wells. Set up three blank wells per 24-well plate with DMSO alone. After addition, seal the wells and incubate in a shaker at 28°C, 180 rpm, in the dark. Incubate for varying times depending on the growth characteristics of different bacteria. After incubation, measure absorbance using a microplate reader. Calculate the inhibition rate based on three replicates.

[0137] 6. Data processing: Corrected OD value = OD value of bacterial culture medium - OD value of sterile culture medium

[0138] Inhibition rate = (corrected OD value of the control culture medium - corrected OD value of the toxic culture medium) / corrected OD value of the control culture medium × 100%.

[0139] Table 3

[0140]

[0141] As can be seen from Table 3, at a concentration of 50 mg / L, most of the thiazole compounds containing cyclohexane urea prepared in Examples 1-22 have a certain inhibitory effect on bacteria such as rice leaf streak, rice bacterial blight, rice wilt, cabbage soft rot and citrus ulcer, and the fungicidal activity of some compounds can be higher than 90%.

[0142] In order to intuitively illustrate the performance and characterization process of the thiazole compounds containing cyclic amine urea prepared by the method of the present invention, the present invention exemplarily provides the identification results of the thiazole compounds containing cyclic amine urea prepared in Examples 1-22, including nuclear magnetic resonance hydrogen spectrum, nuclear magnetic resonance carbon spectrum, nuclear magnetic resonance fluorine spectrum, high-resolution data, ee value of single-configuration compound, appearance properties and melting point, as shown in Table 4.

[0143] Table 4

[0144]

[0145]

[0146]

[0147] The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, various simple variations of the technical solution of the present invention may be made, including combining the various technical features in any other appropriate manner. These simple variations and combinations should also be regarded as disclosed in the present invention and fall within the scope of protection of the present invention.

Claims

1. A thiazole compound containing cyclohexane urea, characterized in that: The compound is a compound represented by formula (1); Formula (1): Wherein, R1 is -R4-R5-O-R6, and R4 is selected from C3-C 15 Azacycloalkyl; R5 is selected from C1-C 10 Alkylene; R6 is selected from substituted or unsubstituted C6-C 20 aryl, or selected from substituted or unsubstituted C3-C 12 Heteroaryl; substituted C6-C 20 Aryl and substituted C3-C 12 The substituents of the heteroaryl group are selected from C1-C 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 At least one of a haloalkyl group and a halogen; R2 is selected from H, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C3-C 20 At least one of the cycloalkyl groups; substituted C1-C 12 Alkyl, substituted C2-C 12 Alkenyl and substituted C3-C 20 The substituent of the cycloalkyl group is selected from at least one of a C1-C6 alkyl group, a C2-C6 alkenyl group, and a C3-C8 cycloalkyl group; Each R3 is independently selected from H, C1-C 12 At least one of an alkyl group, a nitro group and a cyano group.

2. The thiazole compound containing cyclohexane according to claim 1, wherein In the compound represented by formula (1), R4 is selected from C3-C 10 R5 is selected from C1-C6 alkylene; R6 is selected from substituted or unsubstituted C6-C 14 aryl and substituted or unsubstituted C3-C 10 Heteroaryl; substituted C6-C 14 Aryl and substituted C3-C 10 The substituent of the heteroaryl is selected from at least one of a C1-C6 alkyl, a C1-C6 alkoxy, a C1-C6 haloalkyl and a halogen; R2 is selected from H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl and substituted or unsubstituted C3-C 10 At least one of cycloalkyl; substituted C1-C6 alkyl, substituted C2-C6 alkenyl and substituted C3-C 10 The substituent of the cycloalkyl group is selected from at least one of a C1-C3 alkyl group, a C2-C4 alkenyl group, and a C3-C6 cycloalkyl group; Each R3 is independently selected from at least one of H, C1-C8 alkyl, nitro and cyano; Preferably, R4 is selected from at least one of azetidinyl, pyrrolidinyl, piperidinyl, azepanyl, azooctanyl, azanonyl, azadecyl and piperazinyl; R5 is selected from at least one of -CH2-, -CH2-CH2-, -CH2-CH2-CH2-, -CH(CH3)-CH2-, -CH2-CH(CH3)-, -CH2-(CH2)2-CH2-, -C(CH3)2-CH2- and -CH2-C(CH3)2-; R6 is selected from at least one of substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, substituted or unsubstituted pyridyl, substituted or unsubstituted piperidyl and substituted or unsubstituted quinolyl; the substituents of the substituted phenyl, substituted naphthyl, substituted pyridyl, substituted piperidyl and substituted quinolyl are selected from -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -CH2CH2CH2CH3, -CH2CH(CH3)2, -CH(CH3)CH2CH3, -C(CH3) 3, at least one of -OCH3, -OCH2CH3, -OCH2CH2CH3, -OCH(CH3)2, -OCH2CH2CH2CH3, -OCH2CH(CH3)2, -OCH(CH3)CH2CH3, -OC(CH3)3, -CH2F, -CH2Cl, -CH2Br, -CH2I, -CHF2, -CHCl2, -CHBr2, -CHI2, -CF3, -CCl3, -CBr3, -CI3, F, Cl, Br and I; R2 is selected from at least one of H, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -CH2CH2CH2CH3, -CH2CH(CH3)2, -CH(CH3)CH2CH3, -C(CH3)3, -CH2CH2CH2CH2CH3, -CH2CH2CH(CH3)2, -CH2C(CH3)3, -CH(CH3)CH2CH2CH3, -CH2CH2CH2CH2CH2CH3, -CH(CH3)CH2CH2CH2CH3, -CH2CH(CH3)CH2CH2CH3, -CH2CH(CH3)CH2CH2CH3, and -C(CH3)2CH2CH2CH3; Each R3 is independently selected from at least one of H, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -CH2CH2CH2CH3, -CH2CH(CH3)2, -CH(CH3)CH2CH3, -C(CH3)3 and nitro.

3. The thiazole compound containing cyclohexane according to claim 1 or 2, wherein The compound is a compound represented by formula (2); Formula (2): Wherein, n=1-8; Preferably, n=1-5.

4. The thiazole compound containing cyclanilamide according to any one of claims 1 to 3, wherein In the compound represented by formula (2), n=1; Preferably, the compound represented by formula (2) is selected from at least one of the following compounds:

5. The thiazole compound containing cyclanamide according to any one of claims 1 to 3, wherein In the compound represented by formula (2), n=2; Preferably, the compound represented by formula (2) is selected from at least one of the following compounds:

6. The thiazole compound containing cyclohexane according to any one of claims 1 to 3, wherein In the compound represented by formula (2), n=3; Preferably, the compound represented by formula (2) is selected from at least one of the following compounds:

7. A method for preparing a thiazole compound containing cycloamine urea according to any one of claims 1 to 6, wherein: The method includes: S1: The compound represented by formula (A) and the compound represented by formula (B) undergo a first contact reaction to obtain a compound represented by formula (C); S2: subjecting the compound represented by formula (C) to a second contact reaction with the compound represented by formula (D) to prepare the thiazole compound containing cyclic amine urea; wherein each R7 is independently selected from H or nitro, and at least one is nitro; and X is halogen.

8. The preparation method according to claim 7, wherein The conditions of the first contact reaction include: temperature of 0-30°C and time of 0.5-5h; Preferably, the conditions for the second contact reaction include: temperature of 20-40° C. and time of 3-10 h.

9. Use of the thiazole compound containing cyclanilamide according to any one of claims 1 to 6 in preventing and controlling Cryptosporidium; Preferably, the Cryptosporidium is Cryptosporidium containing PFOR enzyme; More preferably, the Cryptosporidium is Cryptosporidium parvum.

10. Use of the thiazole compound containing cyclaminozide according to any one of claims 1 to 6 in controlling plant pathogens; Preferably, the plant pathogen is a plant pathogen containing PFOR enzyme; More preferably, the plant pathogenic bacteria include at least one of Brassica rapa, Xanthophyllomyces oryzae, Rhizoctonia solani, Angular leaf spot of cucumber, Xanthophyllomyces citri, and Psoralea corylifolia of oryzae.

Images