Nitrothiazole compound containing glycine fragment as well as preparation method and application of nitrothiazole compound

Abstract

The invention relates to a nitrothiazole compound containing a glycine fragment, and particularly discloses the nitrothiazole compound containing the glycine fragment as well as a preparation method and application of the nitrothiazole compound. The nitrothiazole compound containing the glycine fragment provided by the invention is a compound as shown in a formula (1), a glycine structure and a nitazoxanide drug molecule are spliced and designed, the glycine fragment is introduced into an amide part in nitazoxanide, a carbon chain structure is prolonged, molecular rigidity is reduced, and the solubility of the compound is improved; the activity of the nitrothiazole compound containing the glycine fragment is further improved. Formula (1): # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a nitrothiazole compound containing a glycine fragment, and in particular to a nitrothiazole compound containing a glycine fragment, a preparation method and an application thereof. Background Art

[0002] Cryptosporidiosis is a foodborne and waterborne infectious disease caused by Cryptosporidium parvum infection, which infects a variety of mammals (J Parasit Dis, 2022, 46(3), 923-939). More than 800 cases of human cryptosporidiosis have been reported each year (Int J Parasitol, 2018, 48(1), 1-12), which has a huge impact worldwide. Currently, more than 40 different Cryptosporidium species have been confirmed, 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, and cases can be found in most terrestrial livestock species except Antarctica, 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] The inventors discovered that nitazoxanide suffers from low solubility, likely due to its relatively simple molecule and rigid structure, which reduces the compound's availability and compromises its activity. They plan to design a splice between the glycine structure and the nitazoxanide drug molecule, introducing a glycine fragment into the amide portion of nitazoxanide. This will help extend the carbon chain structure and reduce molecular rigidity, thereby increasing the compound's solubility and, consequently, its activity. Summary of the Invention

[0005] The purpose of the present invention is to overcome the problems of nitazoxanide resistance, unclear complete structure-activity relationship for Cryptosporidium, poor solubility and activity in the prior art, and to provide a nitrothiazole compound containing a glycine fragment, which has good biological activity and can be used for the prevention and treatment of cryptosporidiosis.

[0006] In order to achieve the above-mentioned object, the present invention provides a nitrothiazole compound containing a glycine fragment, wherein the compound is a compound represented by formula (1);

[0007] Formula (1):

[0008] Wherein, R1 is selected from R4 is selected from C1-C6 alkylene; R5 is selected from substituted or unsubstituted C6-C 20 Aryl, substituted C6-C 20 The substituents of the aryl group are selected from C1-C 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 At least one of a haloalkyl group, a hydroxyl group, and a halogen group;

[0009] R2 is selected from H, substituted or unsubstituted C1-C 15 Alkyl and substituted or unsubstituted C2-C 15 At least one of the alkenyl groups; substituted C1-C 15 Alkyl and substituted C2-C 15 The substituent of the alkenyl group is selected from at least one of a C1-C6 alkyl group, a C2-C6 alkenyl group and a C3-C8 cycloalkyl group;

[0010] Each R3 is independently selected from H, C1-C 12 At least one of an alkyl group, a nitro group and a cyano group.

[0011] The second aspect of the present invention provides a method for preparing the nitrothiazole compound containing a glycine fragment according to the first aspect of the present invention, the method comprising:

[0012] The compound represented by formula (A) is subjected to a first contact reaction with the compound represented by formula (B) to prepare the nitrothiazole compound containing a glycine fragment;

[0013] Formula (A): Formula (B):

[0014] A third aspect of the present invention provides a method for preparing a nitrothiazole compound containing a glycine fragment, the method comprising:

[0015] (1) subjecting the compound represented by formula (A') to a second contact reaction with the compound represented by formula (B) to obtain the compound represented by formula (C);

[0016] (2) subjecting the compound represented by formula (C) to a substitution reaction with the compound represented by formula (D) to prepare the nitrothiazole compound containing a glycine fragment;

[0017] Formula (A'): Formula (B):

[0018] Formula (C): Formula (D): R2-X;

[0019] Wherein, X is a halogen.

[0020] The fourth aspect of the present invention provides a use of the nitrothiazole compound containing a glycine fragment according to the first aspect of the present invention in preventing and controlling Cryptosporidium.

[0021] The fifth aspect of the present invention provides a use of the nitrothiazole compound containing a glycine fragment according to the first aspect of the present invention in preventing and controlling plant pathogens.

[0022] Through the above technical solution, the glycine structure and the nitazoxanide drug molecule are spliced and designed, and a glycine fragment is introduced into the amide part of nitazoxanide, which is beneficial to extend the carbon chain structure and reduce the molecular rigidity to improve the solubility of the compound, and further improve the activity of the nitrothiazole compound containing the glycine fragment. DETAILED DESCRIPTION

[0023] 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.

[0024] In one aspect, the present invention provides a nitrothiazole compound containing a glycine fragment, wherein the compound is a compound represented by formula (1);

[0025] Formula (1):

[0026] Wherein, R1 is selected from R4 is selected from C1-C6 alkylene; R5 is selected from substituted or unsubstituted C6-C 20 Aryl, substituted C6-C 20 The substituents of the aryl group are selected from C1-C 10 Alkyl, C1-C10 Alkoxy, C1-C 10 At least one of a haloalkyl group, a hydroxyl group, and a halogen group;

[0027] R2 is selected from H, substituted or unsubstituted C1-C 15 Alkyl and substituted or unsubstituted C2-C 15 At least one of the alkenyl groups; substituted C1-C 15 Alkyl and substituted C2-C 15 The substituent of the alkenyl group is selected from at least one of a C1-C6 alkyl group, a C2-C6 alkenyl group and a C3-C8 cycloalkyl group;

[0028] Each R3 is independently selected from H, C1-C 12 At least one of an alkyl group, a nitro group and a cyano group.

[0029] In the present invention, in order to further improve the solubility and activity of the nitrothiazole compound containing the glycine fragment, preferably, R4 is selected from a C1-C3 alkylene group; R5 is selected from a substituted or unsubstituted C6-C 15 Aryl, substituted C6-C 15 The substituent of the aryl group is selected from at least one of a C1-C6 alkyl group, a C1-C6 alkoxy group, a C1-C6 haloalkyl group, a hydroxyl group and a halogen group;

[0030] R2 is selected from H, substituted or unsubstituted C1-C 12 Alkyl and substituted or unsubstituted C2-C 12 At least one of the alkenyl groups; substituted C1-C 12 Alkyl and substituted C2-C 12 The substituent of the alkenyl group is selected from at least one of a C1-C6 alkyl group, a C2-C6 alkenyl group and a C3-C8 cycloalkyl group;

[0031] Each R3 is independently selected from at least one of H, a C1-C6 alkyl group, a nitro group, and a cyano group.

[0032] In the present invention, preferably, R4 is selected from at least one of -CH2-, -CH2-CH2-, -CH2-CH2-CH2-, -CH(CH3)-CH2- and -CH2-CH(CH3)-;

[0033] R5 is selected from substituted or unsubstituted phenyl, or from substituted or unsubstituted naphthyl, the substituents of the substituted phenyl and substituted naphthyl being selected from at least one of -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -CH2CH2CH2CH3, -CH2CH(CH3)2, -CH(CH3)CH2CH3, -C(CH3)3, -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, I and hydroxyl;

[0034] R2 is selected from H, substituted or unsubstituted C1-C 10 Alkyl and substituted or unsubstituted C2-C 10 At least one of the alkenyl groups, substituted C1-C 10 Alkyl and substituted C2-C 10 The substituents in the alkenyl group are selected from at least one of cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl;

[0035] 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.

[0036] According to a preferred embodiment, in the compound represented by formula (1), R2 is H.

[0037] In the present invention, preferably, the compound represented by formula (1) is selected from at least one of the following compounds:

[0038] Formula (c1): Formula (c2):

[0039] Formula (c3): Formula (c4):

[0040] Formula (c5): Formula (c6):

[0041] Formula (c7): Formula (c8):

[0042] Formula (c9): Formula (c10):

[0043] Formula (c11): Formula (c12):

[0044] Formula (c13):

[0045] According to another preferred embodiment, in the compound represented by formula (1), R2 is selected from substituted or unsubstituted C1-C8 alkyl or substituted or unsubstituted C2-C8 alkenyl, and the substituents in the substituted C1-C8 alkyl and the substituted C2-C8 alkenyl are selected from at least one of cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0046] In the present invention, preferably, the compound represented by formula (1) is selected from at least one of the following compounds:

[0047] Formula (d1): Formula (d2)

[0048] Formula (d3): Formula (d4):

[0049] The second aspect of the present invention provides a method for preparing the nitrothiazole compound containing a glycine fragment according to the first aspect of the present invention, the method comprising:

[0050] The compound represented by formula (A) is subjected to a first contact reaction with the compound represented by formula (B) to prepare the nitrothiazole compound containing a glycine fragment;

[0051] Formula (A): Formula (B):

[0052] In the present invention, the first contact reaction is carried out by subjecting the amino group in the compound represented by formula (A) to an amidation reaction with the hydroxy acid group in the compound represented by formula (B) to form an amide bond, so as to prepare the compound represented by formula (1), that is, the nitrothiazole compound containing a glycine fragment.

[0053] In the present invention, preferably, the conditions for the first contact reaction include: temperature of 20-40°C and time of 1-6 hours; more preferably, the conditions for the first contact reaction include: temperature of 25-35°C and time of 2-4 hours.

[0054] In the present invention, to improve the efficiency of the first contact reaction, the hydroxy acid groups in the compound represented by formula (B) can be activated using a condensing agent. To this end, the first contact reaction is preferably carried out in the presence of a first condensing agent; the first condensing agent is selected from at least one of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), hydroxybenzotriazole (HOBt), and diisopropylcarbodiimide (DIC). More preferably, the molar ratio of the compound represented by formula (B) to the first condensing agent is 1:1-4, preferably 1:2.5-3.5, and can be, for example, 1:1, 1:2.5, 1:3, 1:3.5, and any range therebetween.

[0055] In the present invention, to neutralize the acidic substances generated in the first contact reaction and simultaneously provide an alkaline environment to facilitate nucleophilic attack of the amino group in the compound represented by formula (A) and promote the formation of an amide bond, the first contact reaction is preferably carried out in a first alkaline reagent, wherein the first alkaline reagent is at least one selected from N,N-diisopropylethylamine (DIEA), triethylamine (EtN), and pyridine. More preferably, the molar ratio of the compound represented by formula (A) to the first alkaline reagent is 1:2-4, preferably 1:2.5-3.5, and can be, for example, 1:1, 1:2.5, 1:3, 1:3.5, and any range therebetween.

[0056] In the present invention, to accelerate the formation of an amide bond between the amino group in the compound represented by formula (A) and the hydroxy acid group in the compound represented by formula (B), the first contact reaction is carried out in the presence of a first catalyst. Preferably, the first catalyst is selected from 4-dimethylaminopyridine (DMAP) and / or 4-pyridylpyridine (4-PPY). More preferably, the molar ratio of the compound represented by formula (A) to the first catalyst is 1:0.05-0.5, preferably 1:0.08-0.15, and can be, for example, 1:0.08, 1:0.1, 1:1.5, and any range therebetween.

[0057] In the present invention, the first contact reaction is preferably carried out in a first solvent, wherein the first solvent is selected from at least one of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), and tetrahydrofuran (THF). More preferably, the mass ratio of the compound represented by formula (A) to the solvent is 1:5-35, preferably 1:15-30, for example, 1:15, 1:20, 1:25, 1:27, 1:35, and any range therebetween.

[0058] In the present invention, preferably, the compound represented by formula (A) is prepared by a condensation reaction and a hydrolysis reaction, comprising: subjecting the compound represented by formula (a) and the compound represented by formula (b) to a condensation reaction to prepare a compound represented by formula (c), and then subjecting the compound represented by formula (c) to a hydrolysis reaction in an acidic reagent to prepare a compound represented by formula (A);

[0059] Formula (a): Formula (b):

[0060] Formula (c)

[0061] In the present invention, the condensation reaction is carried out by connecting the amino group of the compound represented by formula (a) with the carboxylic acid group of the compound represented by formula (b) to form an amide bond, and simultaneously using a tert-butyloxycarbonyl protecting group to protect the amino group to prepare the compound represented by formula (c).

[0062] In the present invention, preferably, the condensation reaction conditions include: temperature of 20-40°C, time of 2-6 hours. More preferably, the condensation reaction conditions include: temperature of 25-35°C, time of 3-5 hours.

[0063] In the present invention, to activate the carboxylic acid group of the compound represented by formula (b), accelerate the formation of the amide bond, and prevent the carboxylic acid esterification side reaction, the condensation reaction is preferably carried out in the presence of a second condensing agent, wherein the second condensing agent is selected from at least one of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), hydroxybenzotriazole (HOBt), and diisopropylcarbodiimide (DIC). More preferably, the molar ratio of the compound represented by formula (b) to the second condensing agent is 1:1-4, preferably 1:1.5-3.5, for example, 1:1.5, 1:3, 1:3.5, and any range therebetween.

[0064] In the present invention, to neutralize the acidic substances in the condensation reaction and provide an alkaline environment to connect the aminocarboxylic acid groups in the compound represented by formula (a) to form an amide bond, the condensation reaction is preferably carried out in the presence of a second alkaline reagent, wherein the second alkaline reagent is selected from at least one of N,N-diisopropylethylamine (DIPEA), triethylamine (EtN), and pyridine. More preferably, the molar ratio of the compound represented by formula (a) to the second alkaline reagent is 1:2-4, preferably 1:1.5-3.5, for example, 1:1.5, 1:3, 1:3.5, and any range therebetween.

[0065] In the present invention, to accelerate the formation of the amide bond in the condensation reaction, a catalyst can be added. Preferably, the condensation reaction is carried out in the presence of a second catalyst. Preferably, the second catalyst is selected from 4-dimethylaminopyridine (DMAP) and / or 4-pyridylpyridine (4-PPY). More preferably, the molar ratio of the compound represented by formula (a) to the second catalyst is 1:0.05-0.5, preferably 1:0.08-0.15, for example, 1:0.08, 1:0.1, 1:0.15, and any range therebetween.

[0066] In the present invention, the condensation reaction is preferably carried out in the presence of a second solvent, wherein the second solvent is selected from at least one of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), and tetrahydrofuran (THF). More preferably, the mass ratio of the compound represented by formula (a) to the solvent is 1:5-30, preferably 1:10-25, for example, 1:10, 1:15, 1:18, 1:20, and any range therebetween.

[0067] In the present invention, preferably, the condensation reaction further includes purifying the reaction solution to prepare a compound shown in formula (c); the purification treatment includes: after the condensation reaction is completed, the reaction solution is sequentially quenched with water, extracted with EA (ethyl acetate), washed with 1M hydrochloric acid solution, washed with saturated NaHCO3 solution, washed with saturated brine, dried with anhydrous sodium sulfate, desolvated under reduced pressure, and slurried with EA-PE (ethyl acetate-petroleum ether) to prepare a compound shown in formula (c).

[0068] In the present invention, the hydrolysis reaction is performed by removing the tert-butyloxycarbonyl protecting group from the compound represented by formula (c) using an acidic reagent to prepare the compound represented by formula (A). Preferably, the acidic reagent is selected from at least one of HCl, H2PO4, and HClO4. More preferably, the molar ratio of the compound represented by formula (c) to the acidic reagent is 1:5-20, preferably 1:8-15, for example, 1:8, 1:10, 1:12, 1:15, and any range therebetween.

[0069] In the present invention, preferably, the conditions for the hydrolysis reaction include: temperature of 20-40°C and time of 6-12 hours; more preferably, the conditions for the hydrolysis reaction include: temperature of 25-35°C and time of 8-11 hours.

[0070] In the present invention, the hydrolysis reaction is preferably carried out in the presence of a third solvent, wherein the third solvent is selected from at least one of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), and tetrahydrofuran (THF). More preferably, the mass ratio of the compound represented by formula (c) to the solvent is 1:5-20, preferably 1:8-15, for example, 1:8, 1:10, 1:12, 1:15, and any range therebetween.

[0071] In the present invention, preferably, the hydrolysis reaction further comprises purifying the reaction solution to prepare the compound represented by formula (A); the purification treatment comprises: after the hydrolysis reaction is completed, desolventizing, adding THF solution for ultrasonic washing, filtering, collecting the filter residue and drying it to prepare the compound represented by formula (A).

[0072] The specific process of the preparation method of the nitrothiazole compound containing a glycine fragment provided in the second aspect of the present invention is as follows:

[0073]

[0074] R1-R3 in the compound represented by the reaction product formula (1) are provided by corresponding groups in the compound represented by formula (a), the compound represented by formula (b), the compound represented by formula (c), the compound represented by formula (A) and the compound represented by formula (B), 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 preparation method of the nitrothiazole compound containing a glycine fragment, and the present invention will not be repeated here.

[0075] In the present invention, preferably, when the compound represented by formula (a) is the compound represented by formula (a-1), and the compound represented by formula (b) is the compound represented by formula (b-1), the specific process of the preparation method of the nitrothiazole compound containing a glycine fragment provided in the second aspect of the present invention is as follows:

[0076]

[0077] A third aspect of the present invention provides a method for preparing a nitrothiazole compound containing a glycine fragment, the method comprising:

[0078] (1) subjecting the compound represented by formula (A') to a second contact reaction with the compound represented by formula (B) to obtain the compound represented by formula (C);

[0079] (2) subjecting the compound represented by formula (C) to a substitution reaction with the compound represented by formula (D) to prepare the nitrothiazole compound containing a glycine fragment;

[0080] Formula (A'): Formula (B):

[0081] Formula (C): Formula (D): R2-X;

[0082] Wherein, X is a halogen.

[0083] The preparation method provided in the third aspect of the present invention is for preparing a nitrothiazole compound containing a glycine fragment when R2 is not H. Preferably, the compound represented by formula (A') is also prepared by the condensation reaction and hydrolysis reaction described in the second aspect of the present invention. Except for using the compound represented by formula (a') instead of the compound represented by formula (a), the remaining reaction raw materials and conditions are consistent with the requirements of the second aspect of the present invention.

[0084] Formula (a'):

[0085] In the present invention, preferably, the second contact reaction is carried out by amidation reaction of the amino group in the compound represented by formula (A') with the hydroxy acid group in the compound represented by formula (B) to form an amide bond to prepare the compound represented by formula (C).

[0086] In the present invention, preferably, the conditions for the second contact reaction include: temperature of 20-40°C and time of 1-6 hours; more preferably, the conditions for the second contact reaction include: temperature of 25-35°C and time of 2.5-4.5 hours.

[0087] In the present invention, to improve the efficiency of the second contact reaction, the hydroxy acid groups in the compound represented by formula (B) can be activated using a condensing agent. To this end, the second contact reaction is preferably carried out in the presence of a third condensing agent; the third condensing agent is selected from at least one of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), hydroxybenzotriazole (HOBt), and diisopropylcarbodiimide (DIC). More preferably, the molar ratio of the compound represented by formula (B) to the third condensing agent is 1:1-4, preferably 1:1.5-3.5, and can be, for example, 1:1.5, 1:2, 1:3.5, and any range therebetween.

[0088] In the present invention, to neutralize the acidic substances generated in the second contact reaction and simultaneously provide an alkaline environment to facilitate nucleophilic attack of the amino group in the compound represented by Formula (A') and promote amide bond formation, the second contact reaction is preferably carried out in the presence of a third alkaline reagent, wherein the third alkaline reagent is at least one selected from N,N-diisopropylethylamine (DIEA), triethylamine (EtN), and pyridine. More preferably, the molar ratio of the compound represented by Formula (A') to the third alkaline reagent is 1:2-4, preferably 1:1.5-3.5, and can be, for example, 1:1.5, 1:2, 1:3.5, and any range therebetween.

[0089] In the present invention, to accelerate the formation of an amide bond between the amino group in the compound represented by formula (A') and the hydroxy acid group in the compound represented by formula (B), the second contact reaction is carried out in the presence of a third catalyst. Preferably, the third catalyst is selected from 4-dimethylaminopyridine (DMAP) and / or 4-pyridylpyridine (4-PPY). More preferably, the molar ratio of the compound represented by formula (A) to the third catalyst is 1:0.05-0.5, preferably 1:0.08-0.15, for example, 1:0.08, 1:0.1, 1:0.15, and any range therebetween.

[0090] In the present invention, the second contact reaction is preferably carried out in a fourth solvent, wherein the fourth solvent is selected from at least one of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), and tetrahydrofuran (THF). More preferably, the mass ratio of the compound represented by formula (A') to the fourth solvent is 1:5-30.

[0091] In the present invention, the substitution reaction is carried out by deprotonating the amino group in the compound represented by formula (C) to generate an amino anion, which then undergoes a nucleophilic substitution reaction with the compound represented by formula (D) to form a new CN bond, thereby preparing the compound represented by formula (1), that is, a nitrothiazole compound containing a glycine fragment.

[0092] In the present invention, preferably, the conditions for the substitution reaction include: temperature of 20-80° C., time of 0.5-4 h. More preferably, the conditions for the substitution reaction include: temperature of 40-60° C., time of 1-3 h.

[0093] In the present invention, the deprotonation of the amino group in the compound represented by formula (C) to form an amino anion requires the participation of an alkaline reagent. Preferably, the substitution reaction is carried out in the presence of a fourth alkaline reagent, wherein the fourth alkaline reagent is selected from at least one of NaH, diisopropylethylamine (DIPEA), and triethylamine (EtN). More preferably, the molar ratio of the compound represented by formula (C) to the fourth alkaline reagent is 1:1-3, preferably 1:1.2-2, and can be, for example, 1:1.2, 1:1.5, 1:2, and any range therebetween.

[0094] In the present invention, preferably, the substitution reaction is carried out in a fifth solvent, wherein the fifth solvent is selected from at least one of dimethylformamide (DMF), dimethyl sulfoxide (DMSO), and tetrahydrofuran (THF). More preferably, the mass ratio of the compound represented by formula (C) to the fifth solvent is 1:5-30.

[0095] The specific process of the preparation method of the nitrothiazole compound containing a glycine fragment provided in the third aspect of the present invention is as follows:

[0096]

[0097] R1-R3 in the compound represented by the reaction product formula (1) are provided by corresponding groups in the compound represented by formula (a'), the compound represented by formula (b), the compound represented by formula (c'), the compound represented by formula (A'), the compound represented by formula (B) and the compound represented by formula (C), 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 preparation method of the nitrothiazole compound containing a glycine fragment, and the present invention will not be repeated here.

[0098] In the present invention, preferably, when the compound represented by formula (a') is the compound represented by formula (a'-1), and the compound represented by formula (b) is the compound represented by formula (b-1), the specific process of the preparation method of the nitrothiazole compound containing a glycine fragment provided by the third aspect of the present invention is as follows:

[0099]

[0100] In the present invention, preferably, the preparation method of the nitrothiazole compound containing a glycine fragment further includes post-treatment of the reaction solution, wherein the post-treatment includes: quenching the reaction solution with water, adding a solution (e.g., tetrahydrofuran), extracting with ethyl acetate, washing the organic phase with a washing liquid (e.g., at least one of a 1M hydrochloric acid solution, a saturated NaHCO3 solution, and a saturated saline solution as a washing liquid), adding anhydrous sodium sulfate for drying, desolvating under reduced pressure, slurrying with ethyl acetate-petroleum ether, filtering, and drying.

[0101] The fourth aspect of the present invention provides a use of the nitrothiazole compound containing a glycine fragment according to the first aspect of the present invention in preventing and controlling Cryptosporidium.

[0102] In the present invention, preferably, the Cryptosporidium is Cryptosporidium containing PFOR enzyme. More preferably, the Cryptosporidium is Cryptosporidium parvum.

[0103] The fifth aspect of the present invention provides a use of the nitrothiazole compound containing a glycine fragment according to the first aspect of the present invention in preventing and controlling plant pathogens.

[0104] In the present invention, preferably, the plant pathogen is a plant pathogen containing PFOR enzyme. More preferably, the plant pathogen includes at least one of Brassica rapa, Xanthomonas oryzae, Ralstonia solani, Cucumber angular leaf spot, Citrus canker, and Psoralea corylifolia.

[0105] The nitrothiazole compounds containing glycine fragments prepared by the above technical scheme have excellent activity. According to some preferred embodiments, the nitrothiazole compounds containing glycine fragments have a certain anti-Cryptosporidium effect, and the inhibition rate of some compounds is higher than 50%. At a concentration of 50 mg / L, most nitrothiazole compounds containing glycine fragments 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 50%.

[0106] The present invention will be described in detail below through examples.

[0107] Examples 1-13

[0108] This example is used to illustrate the preparation method of the compounds represented by formula (c1)-(c13). The specific preparation process is as follows:

[0109]

[0110] The specific preparation method is as follows:

[0111] (1) At room temperature (25°C, the same below), 10 mmol of the compound represented by formula (a), 10 mmol of the compound represented by formula (b), 15 mmol of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 15 mmol of hydroxybenzotriazole (HOBt), 1 mmol of 4-dimethylaminopyridine (DMAP), and 30 mmol of N,N-diisopropylethylamine (DIPEA) were placed in 30 mL of dimethylformamide (DMF), stirred and mixed at 25°C for 4 hours to carry out condensation reaction. After the reaction was completed, water was added to quench the mixture, and the mixture was extracted with EA (ethyl acetate) three times. The mixture was washed with 1 M hydrochloric acid solution, saturated NaHCO3 solution, and saturated brine, respectively, and dried over anhydrous sodium sulfate. After desolvation under reduced pressure, the mixture was slurried with EA-PE (ethyl acetate-petroleum ether) to prepare the compound represented by formula (c);

[0112] (2) At room temperature, 10 mmol of the compound represented by formula (c) and 100 mmol of HCl were placed in 30 mL of tetrahydrofuran (THF), stirred and hydrolyzed at 25°C for 10 h. After the reaction was completed, the solvent was removed under reduced pressure, a THF solution was added for ultrasonic washing, and the filter residue was collected and dried after filtration to prepare the compound represented by formula (A);

[0113] (3) At room temperature, 10 mmol of the compound represented by formula (A), 10 mmol of the compound represented by formula (B), 15 mmol of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 15 mmol of hydroxybenzotriazole (HOBt), 1 mmol of 4-dimethylaminopyridine (DMAP), and 30 mmol of N,N-diisopropylethylamine (DIEA) were placed in 40 mL of dimethylformamide (DMF) and stirred for 3 h to perform a first contact reaction;

[0114] (4) After the reaction of step (3) is completed, the reaction solution is quenched by adding water, 100 mL of tetrahydrofuran (THF) solution is added, and the organic phase is extracted three times with ethyl acetate (EA), washed with 1 M hydrochloric acid solution, saturated NaHCO3 solution, and saturated brine, and then dried over anhydrous sodium sulfate. After desolvation under reduced pressure, EA-PE pulping is performed, and the mixture is filtered and dried to obtain the compound represented by formula (1-1);

[0115] Among them, when the compound represented by formula (B) as shown in Table 1 is used, the corresponding compound represented by formula (1-1) prepared is the compound represented by formula (c1)-(c13), and the yield of step (4) is shown in the following table.

[0116] Table 1

[0117]

[0118]

[0119]

[0120] Examples 14-17

[0121] This example is used to illustrate the preparation method of the compounds represented by formula (d1)-(d4). The specific preparation process is as follows:

[0122]

[0123] The specific preparation method is as follows:

[0124] (1) At room temperature (25°C, the same below), 10 mmol of the compound represented by formula (a'), 10 mmol of the compound represented by formula (b), 15 mmol of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 15 mmol of hydroxybenzotriazole (HOBt), 1 mmol of 4-dimethylaminopyridine (DMAP), and 30 mmol of N,N-diisopropylethylamine (DIPEA) were placed in 30 mL of dimethylformamide (DMF) and stirred for 4 h to carry out a condensation reaction. After the condensation reaction was completed, water was added to quench the mixture, and the mixture was extracted with EA three times. The mixture was washed with 1 M hydrochloric acid solution, saturated NaHCO3 solution, and saturated brine, respectively, and dried with anhydrous sodium sulfate. After desolvation under reduced pressure, the mixture was slurried with EA-PE to prepare the compound represented by formula (c');

[0125] (2) At room temperature, 10 mmol of the compound represented by formula (c') and 100 mmol of HCl were placed in 30 mL of tetrahydrofuran (THF), stirred and mixed for 10 hours for hydrolysis reaction. After the reaction was completed, the solution was removed under reduced pressure, ultrasonically washed with THF solution, filtered, and the residue was collected and dried to prepare the compound represented by formula (A');

[0126] (3) At room temperature, 10 mmol of the compound represented by formula (A'), 10 mmol of the compound represented by formula (B), 15 mmol of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 15 mmol of hydroxybenzotriazole (HOBt), 1 mmol of 4-dimethylaminopyridine (DMAP), and 30 mmol of N,N-diisopropylethylamine (DIEA) were placed in 40 mL of dimethylformamide (DMF), stirred and mixed, and a second contact reaction was carried out for 3 hours to prepare a compound represented by formula (C);

[0127] (4) 1 mmol of the compound represented by formula (C), 1.5 mmol of the compound represented by formula (D), and 1.2 mmol of NaH were placed in 5 mL of dimethylformamide (DMF), stirred, and subjected to substitution reaction at 60°C for 1 h;

[0128] (5) After the reaction in step (4) is completed, the reaction solution is quenched by adding water, extracted three times with ethyl acetate (EA), and the organic phase is washed with saturated brine, then dried over anhydrous sodium sulfate, desolvated under reduced pressure, and purified by column chromatography (PE:EA=4:1), filtered, and dried to obtain compounds represented by formulas (d1) to (d4);

[0129] Among them, when the compound represented by formula (B) and the compound represented by formula (D) as shown in Table 2 are used, the corresponding compounds represented by formula (1-2) prepared are compounds represented by formula (d1)-(d4), and the yield of step (5) is shown in the following table.

[0130] Table 2

[0131]

[0132]

[0133] Test Example 1

[0134] The anti-Cryptosporidium activity of the glycine fragment-containing nitrothiazole compounds prepared in the above examples was determined, while 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 3.

[0135] Test method:

[0136] 1. According to 2×10 4 cells and 10 4 The infection model was established with a dose of oocysts per well.

[0137] 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.

[0138] 3. Extract RNA and perform qPCR analysis of human and Cryptosporidium 18S. Calculate the inhibition rate based on the results of three replicates.

[0139] 4. The remaining compounds were tested against Cryptosporidium according to the above procedure.

[0140] Table 3

[0141] serial number Inhibition rate serial number Inhibition rate serial number Inhibition rate serial number Inhibition rate Formula c1 0 Formula c6 0 Formula c11 32 Formula d3 27 Formula c2 31 Formula c7 45 Formula c12 0 Formula d4 57 Formula c3 54 Formula c8 45 Formula c13 0 Nitazoxanide 42 Formula c4 52 Formula c9 40 Formula d1 41 Formula c5 38 Formula c10 0 Formula d2 0

[0142] As shown in Table 3, most of the nitrothiazole compounds containing glycine fragments prepared in Examples 1-17 have a certain anti-Cryptosporidium effect, and some compounds have an inhibition rate higher than 50%.

[0143] Test Example 2

[0144] The anti-plant pathogenic fungal activity of the nitrothiazole compounds containing glycine fragments 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 nitrothiazole compounds containing glycine fragments and the control group are shown in Table 4.

[0145] Test method:

[0146] 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.

[0147] 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.

[0148] 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.

[0149] 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.

[0150] 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.

[0151] 6. Data processing: Corrected OD value = OD value of bacterial culture medium - OD value of sterile culture medium

[0152] 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%.

[0153] Table 4

[0154]

[0155] As shown in Table 4, at a concentration of 50 mg / L, most of the nitrothiazole compounds containing glycine fragments prepared in Examples 1-17 had a certain inhibitory effect on bacteria such as rice leaf streak, rice bacterial blight, rice wilt, cabbage soft rot and citrus canker, and the fungicidal activity of some compounds was higher than 50%.

[0156] To intuitively illustrate the performance and characterization process of the glycine-containing nitrothiazole compounds prepared by the method of the present invention, the present invention exemplarily provides the identification results of the glycine-containing nitrothiazole compounds prepared in Examples 1-17, including H NMR spectra, C NMR spectra, F NMR spectra, high-resolution data, ee values of single-configuration compounds, appearance properties, and melting points, as shown in Table 5.

[0157] Table 5

[0158]

[0159]

[0160]

[0161] 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 nitrothiazole compound containing a glycine fragment, wherein: The compound is a compound represented by formula (1); Formula (1): Wherein, R1 is selected from R4 is selected from C1-C6 alkylene; R5 is selected from substituted or unsubstituted C6-C 20 Aryl, substituted C6-C 20 The substituents of the aryl group are selected from C1-C 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 At least one of a haloalkyl group, a hydroxyl group, and a halogen group; R2 is selected from H, substituted or unsubstituted C1-C 15 Alkyl and substituted or unsubstituted C2-C 15 At least one of the alkenyl groups; substituted C1-C 15 Alkyl and substituted C2-C 15 The substituent of the alkenyl 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 nitrothiazole compound containing a glycine fragment according to claim 1, wherein R4 is selected from C1-C3 alkylene; R5 is selected from substituted or unsubstituted C6-C 15 Aryl, substituted C6-C 15 The substituent of the aryl group is selected from at least one of a C1-C6 alkyl group, a C1-C6 alkoxy group, a C1-C6 haloalkyl group, a hydroxyl group and a halogen group; R2 is selected from H, substituted or unsubstituted C1-C 12 Alkyl and substituted or unsubstituted C2-C 12 At least one of the alkenyl groups; substituted C1-C 12 Alkyl and substituted C2-C 12 The substituent of the alkenyl 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 at least one of H, C1-C6 alkyl, nitro and cyano; Preferably, R4 is selected from at least one of -CH2-, -CH2-CH2-, -CH2-CH2-CH2-, -CH(CH3)-CH2- and -CH2-CH(CH3)-; R5 is selected from substituted or unsubstituted phenyl, or from substituted or unsubstituted naphthyl, the substituents of the substituted phenyl and substituted naphthyl being selected from at least one of -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -CH2CH2CH2CH3, -CH2CH(CH3)2, -CH(CH3)CH2CH3, -C(CH3)3, -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, I and hydroxyl; R2 is selected from H, substituted or unsubstituted C1-C 10 Alkyl and substituted or unsubstituted C2-C 10 At least one of the alkenyl groups, substituted C1-C 10 Alkyl and substituted C2-C 10 The substituents in the alkenyl group are selected from at least one of cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl; 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 nitrothiazole compound containing a glycine fragment according to claim 1 or 2, wherein In the compound represented by formula (1), R2 is H; Preferably, the compound represented by formula (1) is selected from at least one of the following compounds:

4. The nitrothiazole compound containing a glycine fragment according to claim 1 or 2, wherein In the compound represented by formula (1), R2 is selected from substituted or unsubstituted C1-C8 alkyl or substituted or unsubstituted C2-C8 alkenyl, and the substituents in the substituted C1-C8 alkyl and substituted C2-C8 alkenyl are selected from at least one of cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl; Preferably, the compound represented by formula (1) is selected from at least one of the following compounds:

5. A method for preparing the nitrothiazole compound containing a glycine fragment according to any one of claims 1 to 4, wherein: The method includes: The compound represented by formula (A) is subjected to a first contact reaction with the compound represented by formula (B) to prepare the nitrothiazole compound containing a glycine fragment; 6. The method for preparing a nitrothiazole compound containing a glycine fragment according to claim 5, wherein: The conditions of the first contact reaction include: temperature of 20-40° C. and time of 1-6 h.

7. A method for preparing a nitrothiazole compound containing a glycine fragment, characterized in that: The method includes: (1) subjecting the compound represented by formula (A') to a second contact reaction with the compound represented by formula (B) to obtain the compound represented by formula (C); (2) subjecting the compound represented by formula (C) to a substitution reaction with the compound represented by formula (D) to prepare the nitrothiazole compound containing a glycine fragment; Wherein, X is a halogen.

8. The preparation method according to claim 5 or 6, wherein The conditions of the second contact reaction include: temperature of 20-40°C and time of 1-6h; Preferably, the conditions of the substitution reaction include: temperature of 20-80° C. and time of 0.5-4 h.

9. Use of the nitrothiazole compound containing a glycine fragment according to any one of claims 1 to 4 in controlling Cryptosporidium; Preferably, the Cryptosporidium is Cryptosporidium containing PFOR enzyme; More preferably, the Cryptosporidium is Cryptosporidium parvum.

10. Use of the nitrothiazole compound containing a glycine fragment according to any one of claims 1 to 4 in controlling plant pathogens; Preferably, the plant pathogenic bacteria are plant pathogenic bacteria 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.

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