CN107827815A - Fluoroquinolones aminoderivative and its purposes for preventing and treating citrus disease - Google Patents

Abstract

Citrus bacterial canker disease and brown spot are citrus Common Diseases, and it is few to prevent and treat the medicine of both diseases at present, and all have the defects of certain.The bit amino of FQNS 7 is connected to obtain the compound as shown in Formulas I or Formula II by the present invention with active fragment by attachment structure, and experiment shows, compound of the invention has preventing and treating citrus bacterial canker disease and the effect with brown spot, has extraordinary application prospect.

Description

Fluoroquinolone amino derivatives and their use in preventing and treating citrus diseases

technical field

The invention relates to fluoroquinolone amino derivatives and uses thereof, in particular to the use of fluoroquinolone amino derivatives in preventing and treating citrus canker and citrus brown spot.

Background technique

As the world's largest fruit, citrus has a dominant position in the whole fruit tree industry due to its output and economic value. In recent years, my country's citrus industry has developed rapidly and has gradually become an important pillar industry to promote the development of the country's rural economy and ensure farmers' income. However, citrus fruits are often susceptible to infection by diseases and insect pests during the process of growth, storage and transportation, and produce and accumulate various biotoxins, which seriously affect the yield and quality of citrus, and greatly reduce the economic value of citrus.

Citrus canker is a widespread bacterial disease caused by Xanthomonas axonopodis pv. Hovenia and Kumquat plants. Xac mainly infects citrus leaves, treetops, fruits, prickles, trunks and other above-ground parts, and can cause ulcers surrounded by chlorosis rings and necrosis on the surface of fruits or leaves, water-soaked lesions on leaves, 2-5 mm in diameter, and leaf cork In addition to leaf symptoms, it can also cause fruit shedding, aging of fruit trees, and loss of commercial value of infected fruits.

Citrus brown spot is a fungal disease caused by Alternaria alternata pathotype tangerine. Alternaria citrus mainly infects the leaves, twigs and fruits of citrus, which not only affects the growth and development of citrus, reduces fruit yield, but also reduces fruit quality, seriously affecting the production of citrus. In recent years, citrus brown spot has occurred in Yunnan, Chongqing, Zhejiang, Hunan and other citrus production areas in my country, and the damage has been increasing year by year, which has attracted great attention from all sides. my country is an important center of origin of citrus. At present, the cultivated area and annual output of citrus ranks first in the world, and there are many varieties of citrus, and the probability of infection is relatively high. Among citrus, red tangerine, Ou tangerine, tribute tangerine and ponkan are all susceptible to disease. However, these varieties account for a large proportion in my country's citrus, so the potential danger of brown spot disease to my country's broad-skinned citrus cannot be ignored.

For citrus canker, due to the high cost of prevention and eradication, and the eradication effect is not obvious, at present foreign and domestic mainly rely on prevention, seedlings in non-citrus canker areas, spraying copper preparations and agricultural antibiotics to prevent and reduce canker the spread. Long-term use of copper preparations is prone to drug resistance, heavy metal tolerance through horizontal transfer between pathogens, and accumulation of copper ions in the soil to become potential phytotoxins and affect the environment. Copper preparations are also a hazard to fruits. , especially in hot and dry environments. Recently, the use of streptomycin in agriculture has been banned, and there is a serious shortage of effective control agents against canker bacteria, and new compounds are urgently needed to replace them.

Citrus brown spot disease is a new disease in my country, and there is a lack of corresponding technical reserves and systematic research, especially the lack of research and accumulation on the types of pathogens that cause damage in different production areas, the law of infestation, effective control technologies, and efficient control agents. Therefore, there is no effective and reliable prevention technology at present. Therefore, it is particularly important and urgent to develop high-efficiency control agents for the prevention and treatment of Alternaria citrus brown spot.

Contents of the invention

The object provided by the present invention is to provide a series of compounds with quinolone structure, their racemates, stereoisomers, tautomers, nitrogen oxides or their pharmaceutically acceptable salts and their effects in preventing and treating citrus canker and brown Use in spot disease. The structure of the compound shown in formula I of the present invention is as follows:

Wherein X is selected from: C1-C6 alkyl; C3-C6 cycloalkyl; substituted or unsubstituted C6-C10 aryl, the substituent on the aryl is one or more, independently selected from: halogen; Amino; Hydroxy; C1-C6 Alkyl; C3-C6 Cycloalkyl;

Z is selected from: N or CR ₇ ; R ₇ is selected from H; C1-C6 alkoxy or halogen;

R ₁ and R ₂ are each independently selected from: H; C1-C6 alkyl; C1-C6 haloalkyl; C1-C6 alkoxy; halogen; hydroxyl; amino or cyano;

Q1 is selected from H _; Halogen;

R ₃ or R ₄ are each independently selected from H; C1-C6 alkyl, said C1-C6 alkyl can be optionally substituted by hydroxyl; amino; halogen; cyano; ureido; thiourea;

Or R ₃ , R ₄ and N together form a 5-8 membered heterocycle, which can be optionally substituted by one or more of the following substituents:

Amino; hydroxyl; tert-butoxycarbonyl; benzyl or C1-C6 alkyl, the C1-C6 alkyl can be optionally replaced by hydroxyl; amino;

Halogen; C1-C4 alkoxy substitution;

A ₁ , A ₂ , A ₃ , A ₄ are independently selected from CR ₅ or N, and at least one of A ₁ and A ₂ is N;

R is selected from H; C1 _- C4 alkyl; C1-C4 alkoxy; amino; amido; hydroxyl; halogen; phenyl or benzyl;

R ₆ is 1-4, independently selected from H, C1-C6 alkyl, the C1-C6 alkyl can be optionally replaced by hydroxyl; amino; halogen or

Cyano substitution; C1-C6 alkoxy, the C1-C6 alkoxy may be optionally substituted by halogen;

B1 is selected from S or NH _;

het is a 5-7 membered heteroaromatic ring containing at least 1 nitrogen atom;

R ₈ is selected from H, C1-C6 alkyl;

n and k are integers of 0 to 3, respectively.

The present invention also provides compounds of the structure shown in formula II:

Wherein X is selected from: C1-C6 alkyl; C3-C6 cycloalkyl; substituted or unsubstituted C6-C10 aryl, the substituent on the aryl is one or more, independently selected from: halogen; Amino; Hydroxy; C1-C6 Alkyl; C3-C6 Cycloalkyl;

Z is selected from: N or CR ₇ ; R ₇ is selected from H; C1-C6 alkoxy or halogen;

R ₁ and R ₂ are each independently selected from: H; C1-C6 alkyl; C1-C6 haloalkyl; C1-C6 alkoxy; halogen; hydroxyl; amino or cyano;

R ₈ is selected from H, C1-C6 alkyl;

Q ₂ from or halogen;

het is a 5-7 membered heteroaromatic ring containing at least 1 nitrogen atom;

n and k are respectively integers from 0 to 3;

m is 1 or 2.

In the compound described in the above formula I or formula II, the substituents can be preferably as follows, and independently freely combined:

het from

The 5-8 membered heterocycle is selected from:

X is selected from: methyl; ethyl; cyclopropyl or 4-fluorophenyl; the claim part does not have this sentence and the next sentence

Z is selected from: N or CR ₇ ; R ₇ is selected from H; methoxy; fluorine or chlorine; the present invention also provides compounds with the following structure:

The present invention also provides a pharmaceutical composition, comprising the above compound, its racemate, stereoisomer, tautomer, nitrogen oxide or their pharmaceutically acceptable salts.

The present invention also provides a pharmaceutical preparation, comprising the above-mentioned compound, its racemate, stereoisomer, tautomer, nitrogen oxide or their pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier and/or auxiliary agent.

The present invention also provides a compound medicine, which is characterized by comprising the above-mentioned compound, its racemate, stereoisomer, tautomer, nitrogen oxide or their pharmaceutically acceptable salt and other active ingredients.

The present invention also provides the use of the above compounds, their racemates, stereoisomers, tautomers, nitrogen oxides or their pharmaceutically acceptable salts in the preparation of medicines for preventing and treating citrus-related diseases. The citrus-related disease is preferably citrus canker or citrus brown spot.

The present invention also provides a method for treating citrus-related diseases, comprising applying the above-mentioned compounds of the present invention, their racemates, stereoisomers, tautomers, nitrogen oxides, or their compounds to citrus fruit trees and/or fruits. Agents of pharmaceutically acceptable salts. Definitions and Explanations of Terms:

Unless otherwise stated, the definitions of groups and terms described in the specification and claims of this application can be combined and combined with each other arbitrarily. Such combinations and combined group definitions and compound structures should fall within the scope of the description of the present application.

The term "tautomer" refers to isomers of functional groups resulting from the rapid movement of an atom in a molecule between two positions. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds can exist in two or more interconvertible species. Prototropic tautomers result from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium and attempts to isolate a single tautomer usually result in a mixture whose physicochemical properties are consistent with the mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates; in phenols, the enol form predominates. The present invention encompasses all tautomeric forms of the compounds.

The term "halogen" refers to F, Cl, Br and I. In other words, F, Cl, Br and I can be described as "halogen" in this specification.

The term "C1-C6" is to be understood as preferably denoting a straight or branched saturated monovalent hydrocarbon group having 1 to 6 carbon atoms, for example methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl , isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl base, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethyl Dimethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl butyl group or 1,2-dimethylbutyl group etc. or their isomers.

For the numerical ranges described in the description and claims of the present application, when the numerical range is defined as an "integer", it should be understood as describing the two endpoints of the range and each integer within the range. For example, "any integer of 0-4" should be understood as describing every integer of 0, 1, 2, 3, and 4.

The pharmaceutically acceptable salts of the compounds mentioned in the present invention may be acid salts or basic salts. Pharmaceutically acceptable salts may be, for example, acid addition salts of sufficiently basic compounds of the invention having nitrogen atoms in the chain or ring, such as acid addition salts with inorganic acids such as hydrochloric acid, hydrofluoric acid, Acids, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric acid, phosphoric acid or nitric acid, or hydrogen sulfate, or acid addition salts with organic acids such as formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid , propionic acid, butyric acid, caproic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)benzoic acid, camphoric acid, cinnamic acid, cyclopentane Propionic acid, gluconic acid, 3-hydroxy-2-naphthoic acid, niacin, paciic acid, pectinic acid, persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid , itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalene disulfonic acid, camphor Sulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, Ascorbic acid, glucoheptanoic acid, glycerophosphate, aspartic acid, sulfosalicylic acid, hemisulfuric acid, or thiocyanic acid.

In addition, another suitable pharmaceutically acceptable salt of a compound of the invention which is sufficiently acidic is an alkali metal salt (such as a sodium or potassium salt), an alkaline earth metal salt (such as a calcium or magnesium salt), an ammonium salt, Or salts formed with organic bases providing physiologically acceptable cations, such as salts formed with sodium ions, potassium ions, N-methylglucamine, dimethylglucamine, ethylglucamine, Lysine, Dicyclohexylamine, Hexamethylenediamine, Ethanolamine, Glucosamine, Meglumine, Sarcosine, Serinol, Trishydroxymethylaminomethane, Aminopropylene Glycol, 1-Amino-2 ,3,4-Butanetriol.

Detailed ways

Example 1

1mmol of gatifloxacin (GAT), 2.5mmol of NaHCO ₃ , and 5mL of DCM were successively added into a 100mL round bottom flask, and a mixed solution of 2mmol of chloroacetyl chloride and 2mL of DCM was added dropwise in an ice bath. After dropping, the reaction was continuously stirred under ice bath, and the reaction progress was monitored by TLC. After the reaction, add a little ice-cold saturated NaCl solution to dissolve the solid, adjust the pH to 4-5 with ice-cold 2N HCl solution, stir evenly, transfer to a separatory funnel for liquid separation, extract twice with DCM, combine the organic phases, and wash with saturated NaCl solution. Dry over Na ₂ SO ₄ , and remove the solvent by rotary evaporation. The pure product was obtained by recrystallization or column chromatography, dried and weighed to obtain the product, [M+H] ⁺ 453.1342.

Example 2

1 mmol of GAT, 2.5 mmol of NaHCO ₃ , and 5 mL of DCM were sequentially added to a 100 mL round bottom flask, and a mixed solution of 2 mmol of chloropropionyl chloride and 2 mL of DCM was added dropwise in an ice bath. After dropping, the reaction was continuously stirred under ice bath, and the reaction progress was monitored by TLC. After the reaction, add a little ice-cold saturated NaCl solution to dissolve the solid, adjust the pH to 4-5 with ice-cold 2N HCl solution, stir evenly, transfer to a separatory funnel for liquid separation, extract twice with DCM, combine the organic phases, and wash with saturated NaCl solution. Dry over Na ₂ SO ₄ , and remove the solvent by rotary evaporation. The pure product was obtained by recrystallization or column chromatography, dried and weighed to obtain the product, [M+H] ⁺ 467.1453.

Example 3

1 mmol of GAT, 2.5 mmol of NaHCO ₃ , and 4 mL of DCM were successively added to a 100 mL round bottom flask, and a mixed solution of 2.5 mmol of 4-chlorobutyryl chloride and 2 mL of DCM was added dropwise in an ice bath. After dropping, the reaction was continuously stirred under ice bath, and the reaction progress was monitored by TLC. After the reaction, add a little ice-cold saturated NaCl solution to dissolve the solid, adjust the pH to 4-5 with ice-cold 2N HCl solution, stir evenly, transfer to a separatory funnel for liquid separation, extract twice with DCM, combine the organic phases, and wash with saturated NaCl solution. Dry over Na ₂ SO ₄ , and remove the solvent by rotary evaporation. The pure product was obtained by recrystallization, dried and weighed to obtain the product, [M+H] ⁺ 481.1679.

Example 4

1.880g (5mmol) of GAT, 20mL of DCM, and 0.601g (6mmol) of succinic anhydride were successively added to a 100mL round bottom flask. After 30 min Na ₂ CO ₃ 0.635 g (6 mmol) was added. The reaction was stirred at room temperature, and the progress of the reaction was monitored by TLC. After the reaction, there was a large amount of yellow oil at the bottom of the reaction bottle. The solvent DCM was removed by rotary evaporation, and 20 mL of H ₂ O was added to dissolve the insoluble matter. Use 2N HCl solution to adjust the pH value to 2, and a large amount of yellow oil appears at the bottom of the bottle. Put it in the refrigerator to refrigerate, and the oil turns into a solid. Suction filtration yields a yellow solid. Natural drying product, [M+H] ⁺ 476.1834.

Example 5

Add 1mmol sarafloxacin and 2mL DCM to a 100mL round bottom flask, cool in an ice bath, stir magnetically, add 3mmolNaHCO ₃ , after 20min, add 2.5mmol 4-chlorobutyryl chloride solution in DCM (2mL) dropwise with a constant pressure dropping funnel ( The dropping speed is about 1d/2s), and the reaction is continued under ice bath after dropping, followed by TLC monitoring until the end of the reaction. Stop stirring, add H ₂ O 15mL and DCM 20mL, adjust pH=3-4 with 1N HCl solution under stirring, if there is solid, let stand, filter with suction, wash the filter cake with DCM 3 times, and keep the filter cake for further purification; the filtrate was transferred to Separating funnel, separation, the aqueous phase was extracted with DCM (15mL×1), the organic phases were combined, washed with saturated NaCl solution (15mL×1), the organic phase was collected, and dried over anhydrous Na ₂ SO ₄ . The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, and dried in vacuum to obtain the product, [M+H] ⁺ 491.1238.

Example 6

Add 1mmol Clinfloxacin and 2mL DCM to a 100mL round bottom flask, cool in an ice bath, stir magnetically, add 3mmolNaHCO ₃ , after 20min, add 2.5mmol 2-chloroacetyl chloride solution in DCM (2mL) dropwise with a constant pressure dropping funnel (drop The acceleration is about 1d/2s), and the reaction was continued under the ice bath after dropping, followed and monitored by TLC until the end of the reaction. Stop stirring, add H ₂ O 15mL and DCM 20mL, adjust pH=3-4 with 1N HCl solution under stirring, if there is solid, let stand, filter with suction, wash the filter cake with DCM 3 times, and keep the filter cake for further purification; the filtrate was transferred to Separating funnel, separation, the aqueous phase was extracted with DCM (15mL×1), the organic phases were combined, washed with saturated NaCl solution (15mL×1), the organic phase was collected, and dried over anhydrous Na ₂ SO ₄ . The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, and dried in vacuum to obtain the product, [M+H] ⁺ 443.0635.

Example 7

Add 1mmol Clinfloxacin and 2mL DCM to a 100mL round bottom flask, cool in an ice bath, stir magnetically, add 3mmolNaHCO ₃ , after 20min, add 2.5mmol 3-chloropropionyl chloride solution in DCM (2mL) dropwise with a constant pressure dropping funnel (drop The acceleration is about 1d/2s), and the reaction was continued under the ice bath after dropping, followed and monitored by TLC until the end of the reaction. Stop stirring, add H ₂ O 15mL and DCM 20mL, adjust pH=3-4 with 1N HCl solution under stirring, if there is solid, let stand, filter with suction, wash the filter cake 3 times with DCM, keep the filter cake for further purification; Transfer to a separatory funnel, separate the layers, extract the aqueous phase with DCM (15 mL×1), combine the organic phases, wash with saturated NaCl solution (15 mL×1), collect the organic phases, and dry over anhydrous Na ₂ SO ₄ . The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, and dried in vacuum to obtain the product, [M+H] ⁺ 457.0814.

Example 8

Add 1mmol Clinfloxacin and 2mL DCM to a 100mL round bottom flask, cool in an ice bath, stir magnetically, add 3mmolNaHCO ₃ , after 20min, add 2.5mmol 4-chlorobutyryl chloride solution in DCM (2mL) dropwise with a constant pressure dropping funnel (drop The acceleration is about 1d/2s), and the reaction was continued under the ice bath after dropping, followed and monitored by TLC until the end of the reaction. Stop stirring, add H ₂ O 15mL and DCM 20mL, adjust pH=3-4 with 1N HCl solution under stirring, if there is solid, let stand, filter with suction, wash the filter cake with DCM 3 times, and keep the filter cake for further purification; the filtrate was transferred to Separating funnel, separation, the aqueous phase was extracted with DCM (15mL×1), the organic phases were combined, washed with saturated NaCl solution (15mL×1), the organic phase was collected, and dried over anhydrous Na ₂ SO ₄ . The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, and dried in vacuum to obtain the product, [M+H] ⁺ 471.0927.

Example 9

Add 1mmol norfloxacin and 2mL DCM to a 100mL round bottom flask, cool in an ice bath, stir magnetically, add 3mmolNaHCO ₃ , after 20min, add 2.5mmol 2-chloroacetyl chloride solution in DCM (2mL) dropwise with a constant pressure dropping funnel (The drop rate is about 1d/2s), and the reaction was continued in an ice bath after the drop was completed, followed by TLC monitoring until the end of the reaction. Stop stirring, add H ₂ O 15mL and DCM 20mL, adjust pH=3-4 with 1N HCl solution under stirring, if there is solid, let stand, filter with suction, wash the filter cake 3 times with DCM, keep the filter cake for further purification; Transfer to a separatory funnel, separate the layers, extract the aqueous phase with DCM (15 mL×1), combine the organic phases, wash with saturated NaCl solution (15 mL×1), collect the organic phases, and dry over anhydrous Na ₂ SO ₄ . The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, and dried in vacuum to obtain the product, [M+H] ⁺ 397.1021.

Example 10

Add 1mmol banoxacin and 2mL DCM to a 100mL round bottom flask, cool in an ice bath, stir magnetically, add 3mmolNaHCO ₃ , after 20min, add 2.5mmol 2-chloroacetyl chloride solution in DCM (2mL) dropwise with a constant pressure dropping funnel (The drop rate is about 1d/2s), and the reaction was continued in an ice bath after the drop was completed, followed by TLC monitoring until the end of the reaction. Stop stirring, add H ₂ O 15mL and DCM 20mL, adjust pH=3-4 with 1N HCl solution under stirring, if there is solid, let stand, filter with suction, wash the filter cake with DCM 3 times, and keep the filter cake for further purification; the filtrate was transferred to Separating funnel, separation, the aqueous phase was extracted with DCM (15mL×1), the organic phases were combined, washed with saturated NaCl solution (15mL×1), the organic phase was collected, and dried over anhydrous Na ₂ SO ₄ . The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, and dried in vacuum to obtain the product, [M+H] ⁺ 467.1428.

Example 11

Add 1mmol sarafloxacin and 2mL DCM to a 100mL round bottom flask, cool in an ice bath, stir magnetically, add 3mmolNaHCO ₃ , after 20min, add 2.5mmol 3-chloropropionyl chloride solution in DCM (2mL) dropwise with a constant pressure dropping funnel ( The dropping speed is about 1d/2s), and the reaction is continued under ice bath after dropping, followed by TLC monitoring until the end of the reaction. Stop stirring, add H ₂ O 15mL and DCM 20mL, adjust pH=3-4 with 1N HCl solution under stirring, if there is solid, let stand, filter with suction, wash the filter cake with DCM 3 times, and keep the filter cake for further purification; the filtrate was transferred to Separating funnel, separation, the aqueous phase was extracted with DCM (15mL×1), the organic phases were combined, washed with saturated NaCl solution (15mL×1), the organic phase was collected, and dried over anhydrous Na ₂ SO ₄ . The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, and dried in vacuum to obtain the product, [M+H] ⁺ 477.1173.

Example 12

To a 100mL round bottom flask, add 2mmol, CHCl ₃ 3mL, K ₂ CO ₃ 2mmol, stirred at room temperature for 30min. Add 1 mmol of intermediate compound Example 1, and move to 35°C water bath for reaction, TLC monitors the reaction progress. After the reaction, 15 mL of DCM, 10 mL of H ₂ O, and 1N HCl solution were added to adjust the pH value to 7-8, and the liquids were separated. Wash with 10 mL of saturated NaCl solution, separate the layers, dry the organic phase over anhydrous Na ₂ SO ₄ , and spin evaporate to obtain a crude product, and column chromatography to obtain a pure product, [M+H] ⁺ 521.2406.

Example 13

1mmol of 2-aminoethanol, 3mL of CHCl ₃ , and 1.2mmol of the compound of Example 3 were added to a 100mL round-bottomed flask in sequence, and the mixture was placed in a 60°C water bath for reflux reaction, and the reaction progress was monitored by TLC. When the compound of Example 3 was no longer reduced, the reaction was stopped. The solvent was removed by rotary evaporation, and the pure product was obtained by column chromatography, [M+H] ⁺ 505.2466.

Example 14

To a 100mL round bottom flask, add 1mmol, CHCl ₃ 3mL, 1.2mmol of the compound of Example 3 was added, and moved to reflux reaction in a water bath at 60°C, and the reaction progress was monitored by TLC. When the compound of Example 3 was no longer reduced, the reaction was stopped. The solvent was removed by rotary evaporation, and the pure product was obtained by column chromatography, [M+H] ⁺ 549.2726.

Example 15

Add 2-aminoacetic acid (50mmol) and methanol 100mL to the reaction bottle, slowly add SOCl ₂ (125mmol) dropwise under ice bath, after the drop is complete, transfer to a 60°C water bath to stir and reflux for reaction, monitor by TLC until the reaction is complete. The methanol and most of the SOCl ₂ were removed by rotary evaporation under reduced pressure, 20 mL of methanol was added, the rotary evaporation was repeated to remove as much SOCl ₂ as possible, and the corresponding methyl 2-aminoacetate hydrochloride was obtained by vacuum drying.

Add INA (20mmol of isonicotinic acid), HOBt (24mmol), DCC (24mmol), DCM (20mL) in turn into the reaction flask, stir for 10min, then add Et ₃ N (60mmol), after 0.5~1h, add 2-methylaminoacetate Ester hydrochloride (22 mmol), TLC monitoring until the end of the reaction. Put it in the refrigerator for 2h, filter with suction, wash the filter cake with DCM, wash the filtrate with a saturated Na ₂ CO ₃ aqueous solution (20mL×2), extract the aqueous phase with DCM (30mL×3), combine the organic phases, anhydrous Na ₂ SO ₄ was dried for 1 h, spin-dried by a rotary evaporator, and column chromatographed to obtain methyl 4-pyridylcarbonylaminoacetate.

Add methyl 4-pyridylcarbonylaminoacetate (15mmol), CH ₃ OH-H ₂ O (V _CH3OH :V _H2O = 3:1) solution (15mL) in sequence in the reaction flask, add LiOH·H ₂ under ice-cooling O (45mmol), stirring, TLC monitoring until the end of the reaction. Remove CH ₃ OH with a rotary evaporator, adjust the pH to about 3 with 4N HCl solution in an ice bath, a large amount of white solid precipitates, filter with suction, wash the filter cake twice with the filtrate and twice with acetone, and dry at 50°C to obtain 4-pyridylcarbonylaminoacetic acid.

Add 4-pyridylcarbonylaminoacetic acid (1mmol), HBTU/TBTU (1.2mmol), DCM (3mL), Et ₃ N (3mmol), and Clinfloxacin (1mmol) successively into the reaction flask, stir under temperature control, and monitor to The reaction is over. Add 20 mL of DCM, transfer to a separatory funnel, wash with saturated Na ₂ CO ₃ aqueous solution (20 mL×1), 0.5N HCl solution (20 mL×1), saturated brine (20 mL×1), and dry over anhydrous Na ₂ SO ₄ After 1h, it was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 528.1445.

Example 16

Add 1mmol of the compound of formula 9, 5mL toluene, 2mmol Et ₃ N into a 100mL round bottom flask, stir for 20-30min, add 2mmol INA, and after 10min, move to 110℃～120℃ oil bath for reflux reaction, monitor by TLC until the reaction is complete . Spin the reaction solution to dryness with a rotary evaporator, add 30mL DCM and stir to dissolve, filter with suction, wash the filter cake 3 times with DCM, wash the filtrate with 10% citric acid solution (15mL×1), collect the organic phase, anhydrous _Na2SO4 dry _. The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 483.1642.

Example 17

Add 1mmol norfloxacin and 2mL DCM to a 100mL round bottom flask, cool in an ice bath, stir magnetically, add 3mmolNaHCO ₃ , after 20min, add 2.5mmol 4-chlorobutyryl chloride solution in DCM (2mL) dropwise with a constant pressure dropping funnel (The drop rate is about 1d/2s), and the reaction was continued in an ice bath after the drop was completed, followed by TLC monitoring until the end of the reaction. Stop stirring, add H ₂ O 15mL and DCM 20mL, adjust pH=3-4 with 1N HCl solution under stirring, if there is solid, let stand, filter with suction, wash the filter cake with DCM 3 times, and keep the filter cake for further purification; the filtrate was transferred to Separating funnel, separation, the aqueous phase was extracted with DCM (15mL×1), the organic phases were combined, washed with saturated NaCl solution (15mL×1), the organic phase was collected, and dried over anhydrous Na ₂ SO ₄ . The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, and the intermediate was obtained by vacuum drying.

Add 1 mmol of the above-mentioned intermediate, 5 mL of toluene, and 2 mmol of Et ₃ N into a 100 mL round bottom flask, stir for 20-30 min, add 2 mmol of INA, and after 10 min, transfer to an oil bath at 110°C to 120°C for reflux reaction, and monitor by TLC until the reaction is complete. Spin the reaction solution to dryness with a rotary evaporator, add 30mL DCM and stir to dissolve, filter with suction, wash the filter cake 3 times with DCM, wash the filtrate with 10% citric acid solution (15mL×1), collect the organic phase, anhydrous _Na2SO4 dry _. The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 411.1956.

Example 18

Add 1mmol enoxacin and 2mL DCM to a 100mL round bottom flask, cool in an ice bath, stir magnetically, add 3mmolNaHCO ₃ , after 20min, add 2.5mmol 2-chloroacetyl chloride DCM (2mL) dropwise with a constant pressure dropping funnel Solution (dropping speed is about 1d/2s), continued to react under ice bath after dropping, TLC tracking monitoring until the end of the reaction. Stop stirring, add H ₂ O 15mL and DCM 20mL, adjust pH=3-4 with 1N HCl solution under stirring, if there is solid, let stand, filter with suction, wash the filter cake 3 times with DCM, and keep the filter cake for further purification; filtrate Move to a separatory funnel, separate the layers, extract the aqueous phase with DCM (15 mL×1), combine the organic phases, wash with saturated NaCl solution (15 mL×1), collect the organic phases, and dry over anhydrous Na ₂ SO ₄ . The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, and the intermediate was obtained by vacuum drying.

Add 1 mmol of the above-mentioned intermediate, 5 mL of toluene, and 2 mmol of Et ₃ N into a 100 mL round bottom flask, stir for 20-30 min, add 2 mmol of INA, and after 10 min, transfer to an oil bath at 110°C to 120°C for reflux reaction, and monitor by TLC until the reaction is complete. Spin the reaction solution to dryness with a rotary evaporator, add 30mL DCM and stir to dissolve, filter with suction, wash the filter cake 3 times with DCM, wash the filtrate with 10% citric acid solution (15mL×1), collect the organic phase, anhydrous _Na2SO4 dry _. The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 484.1616.

Example 19

Add 1mmol enoxacin and 2mL DCM to a 100mL round bottom flask, cool in an ice bath, stir magnetically, add 3mmolNaHCO ₃ , after 20min, add 2.5mmol 4-chlorobutyryl chloride in DCM (2mL) dropwise with a constant pressure dropping funnel Solution (dropping speed is about 1d/2s), continued to react under ice bath after dropping, TLC tracking monitoring until the end of the reaction. Stop stirring, add H ₂ O 15mL and DCM 20mL, adjust pH=3-4 with 1N HCl solution under stirring, if there is solid, let stand, filter with suction, wash the filter cake 3 times with DCM, and keep the filter cake for further purification; filtrate Move to a separatory funnel, separate the layers, extract the aqueous phase with DCM (15 mL×1), combine the organic phases, wash with saturated NaCl solution (15 mL×1), collect the organic phases, and dry over anhydrous Na ₂ SO ₄ . The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, and the intermediate was obtained by vacuum drying.

Add 1mmol of the above-mentioned intermediate, 5mL toluene, and 2mmol Et ₃ N into a 100mL round bottom flask, stir for 20-30min, add 2mmol pyrazinecarboxylic acid (POA), after 10min, move to 110℃～120℃ oil bath for reflux reaction, TLC Monitor until the reaction is complete. Spin the reaction solution to dryness with a rotary evaporator, add 30mL DCM and stir to dissolve, filter with suction, wash the filter cake 3 times with DCM, wash the filtrate with 10% citric acid solution (15mL×1), collect the organic phase, anhydrous _Na2SO4 dry _. The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 513.1892.

Example 20

Add 1 mmol of the compound of Example 10, 5 mL of toluene, and 2 mmol of Et ₃ N into a 100 mL round-bottomed flask, stir for 20-30 min, add 2 mmol of pyrazinecarboxylic acid (POA), and transfer it to an oil bath at 110 ° C to 120 ° C for reflux reaction after 10 min. TLC monitored until the reaction was complete. Spin the reaction solution to dryness with a rotary evaporator, add 30mL DCM and stir to dissolve, filter with suction, wash the filter cake 3 times with DCM, wash the filtrate with 10% citric acid solution (15mL×1), collect the organic phase, anhydrous _Na2SO4 dry _. The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 554.2046.

Example 21

Add 1mmol banoxacin and 2mL DCM to a 100mL round-bottomed flask, cool in an ice bath, stir magnetically, add 3mmolNaHCO ₃ , after 20min, add 2.5mmol 4-chlorobutyryl chloride in DCM (2mL) dropwise with a constant pressure dropping funnel Solution (dropping speed is about 1d/2s), continued to react under ice bath after dropping, TLC tracking monitoring until the end of the reaction. Stop stirring, add H ₂ O 15mL and DCM 20mL, adjust pH=3-4 with 1N HCl solution under stirring, if there is solid, let stand, filter with suction, wash the filter cake 3 times with DCM, and keep the filter cake for further purification; filtrate Move to a separatory funnel, separate the layers, extract the aqueous phase with DCM (15 mL×1), combine the organic phases, wash with saturated NaCl solution (15 mL×1), collect the organic phases, and dry over anhydrous Na ₂ SO ₄ . The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, and the intermediate was obtained by vacuum drying.

Add 1mmol of the above-mentioned intermediate, 5mL toluene, and 2mmol Et ₃ N into a 100mL round bottom flask, stir for 20-30min, add 2mmol pyrazinecarboxylic acid (POA), after 10min, move to 110℃～120℃ oil bath for reflux reaction, TLC Monitor until the reaction is complete. Spin the reaction solution to dryness with a rotary evaporator, add 30mL DCM and stir to dissolve, filter with suction, wash the filter cake 3 times with DCM, wash the filtrate with 10% citric acid solution (15mL×1), collect the organic phase, anhydrous _Na2SO4 dry _. The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 582.2359.

Example 22

Add 2mmol isoniazid, 4mmol NaHCO ₃ and 5mL DCM to a 100mL round-bottom flask in sequence, and after stirring for 20min at -5°C, add 3mmol chloroacetyl chloride solution dropwise with a constant pressure dropping funnel (the drop rate is about 1d/2s) , The reaction was continued at -5°C, followed by TLC monitoring until the end of the reaction. While stirring, add DCM-CH ₃ OH solution (V _DCM : V CH _3OH = 2: 1) until the solids in the reaction flask no longer decrease, let stand, filter with suction, and wash the filter cake 3 times with DCM-CH ₃ OH solution, The filtrate was dried with anhydrous Na ₂ SO ₄ , spin-dried with a rotary evaporator to obtain the crude product, then added 5 mL of DCM and stirred for 20 min, filtered with suction, the filter cake was washed 3 times with DCM, and dried in vacuo to obtain an intermediate for use.

Add 1mmol norfloxacin, 4mmol Et ₃ N and 2mL DMF to a 100mL round-bottomed flask in turn, stir for 20min, add the above intermediate, move to 60°C for 10min and stir for reaction, monitor by TLC until the reaction is complete. Add 10 mL of ice-cold saturated saline solution while stirring, adjust the pH to about 8 with 1N HCl solution, filter with suction, extract the filtrate with DCM (10 mL×2), combine the organic phases, dry with anhydrous Na ₂ SO ₄ , and use a rotary evaporator Spin dry, and purify by column chromatography together with the filter cake to obtain the product, [M+H] ⁺ 497.1943.

Example 23

Add alanine (50mmol) and methanol 100mL to the reaction flask, slowly add SOCl ₂ (125mmol) dropwise under ice bath, after the drop is complete, transfer to a 60°C water bath to stir and reflux for reaction, monitor by TLC until the reaction is complete. The methanol and most of the SOCl ₂ were removed by rotary evaporation under reduced pressure, 20 mL of methanol was added, the rotary evaporation was repeated to remove as much SOCl ₂ as possible, and the corresponding alanine methyl ester hydrochloride was obtained by vacuum drying.

Add INA (20mmol), HOBt (24mmol), DCC (24mmol), DCM (20mL) in turn to the reaction flask, stir for 10min, add DIPEA under ice bath, add alanine methyl ester hydrochloride (22mmol ), TLC monitoring until the end of the reaction. Put it in the refrigerator for 2h, filter with suction, wash the filter cake with DCM, wash the filtrate with a saturated Na ₂ CO ₃ aqueous solution (20mL×2), extract the aqueous phase with DCM (30mL×3), combine the organic phases, anhydrous Na ₂ SO ₄ was dried for 1 h, spin-dried by a rotary evaporator, and the intermediate was obtained by column chromatography.

Add the intermediate (15mmol), CH ₃ OH-H ₂ O (V _CH3OH :V _H2O = 3:1) solution (15mL) to the reaction flask in turn, add LiOH·H ₂ O (45mmol) under ice-cooling, stir, TLC monitoring until the end of the reaction. Remove CH ₃ OH with a rotary evaporator, adjust the pH to about 3 with 4N HCl solution in an ice bath, a large amount of white solid precipitates, filter with suction, wash the filter cake twice with the filtrate and twice with acetone, and dry at 50°C to obtain intermediate.

Add the above-mentioned intermediate compound (1mmol), HBTU/TBTU (1.2mmol), DCM (3mL) in sequence in the reaction flask, add DIPEA3mmol, gatifloxacin (1mmol) under ice bath, stir under temperature control, TLC monitoring until the end of the reaction . Add 20 mL of DCM, transfer to a separatory funnel, wash with saturated Na ₂ CO ₃ aqueous solution (20 mL×1), 0.5N HCl solution (20 mL×1), saturated brine (20 mL×1), and dry with anhydrous Na ₂ SO ₄ for 1 h , spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 552.2217.

Example 24

Add 3-aminopropionic acid (50mmol) and methanol 100mL to the reaction flask, slowly add SOCl ₂ (125mmol) dropwise under ice bath, after the drop is complete, transfer to a 60°C water bath to stir and reflux for reaction, monitor by TLC until the reaction is complete. The methanol and most of the SOCl ₂ were removed by rotary evaporation under reduced pressure, 20 mL of methanol was added, the rotary evaporation was repeated to remove as much SOCl ₂ as possible, and the corresponding methyl 3-aminopropionate hydrochloride was obtained by vacuum drying.

Add INA (20mmol), HOBt (24mmol), DCC (24mmol), DCM (20mL) in turn into the reaction flask, stir for 10min, then add DIPEA under ice bath, add 3-alanine methyl ester hydrochloride after 0.5~1h (22mmol), TLC monitoring until the end of the reaction. Put it in the refrigerator for 2h, filter with suction, wash the filter cake with DCM, wash the filtrate with a saturated Na ₂ CO ₃ aqueous solution (20mL×2), extract the aqueous phase with DCM (30mL×3), combine the organic phases, anhydrous Na ₂ SO ₄ was dried for 1 h, spin-dried by a rotary evaporator, and the intermediate was obtained by column chromatography.

Add the intermediate (15mmol), CH ₃ OH-H ₂ O (V _CH3OH :V _H2O = 3:1) solution (15mL) to the reaction flask in turn, add LiOH·H ₂ O (45mmol) under ice-cooling, stir, TLC monitoring until the end of the reaction. Remove CH ₃ OH with a rotary evaporator, adjust the pH to about 3 with 4N HCl solution in an ice bath, a large amount of white solid precipitates, filter with suction, wash the filter cake twice with the filtrate and twice with acetone, and dry at 50°C to obtain intermediate.

Add the above-mentioned intermediate compound (1mmol), HBTU/TBTU (1.2mmol), DCM (3mL) in sequence in the reaction flask, add DIPEA3mmol, gatifloxacin (1mmol) under ice bath, stir under temperature control, TLC monitoring until the end of the reaction . Add 20 mL of DCM, transfer to a separatory funnel, wash with saturated Na ₂ CO ₃ aqueous solution (20 mL×1), 0.5N HCl solution (20 mL×1), saturated brine (20 mL×1), and dry with anhydrous Na ₂ SO ₄ for 1 h , spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 552.2253.

Example 25

Add 4-aminobutyric acid (50mmol) and 100mL of methanol into the reaction bottle, slowly add SOCl ₂ (125mmol) dropwise under ice bath, after the drop is complete, transfer to a 60°C water bath to stir and reflux for reaction, monitor by TLC until the reaction is complete. The methanol and most of the SOCl ₂ were removed by rotary evaporation under reduced pressure, 20 mL of methanol was added, the rotary evaporation was repeated to remove as much SOCl ₂ as possible, and the corresponding 4-aminobutyric acid methyl ester hydrochloride was obtained by vacuum drying.

Add INA (20mmol), HOBt (24mmol), DCC (24mmol), DCM (20mL) in turn into the reaction flask, stir for 10min, add DIPEA under ice bath, add 4-aminobutyric acid methyl ester hydrochloride after 0.5~1h (22mmol), TLC monitoring until the end of the reaction. Put it in the refrigerator for 2h, filter with suction, wash the filter cake with DCM, wash the filtrate with a saturated Na ₂ CO ₃ aqueous solution (20mL×2), extract the aqueous phase with DCM (30mL×3), combine the organic phases, anhydrous Na ₂ SO ₄ was dried for 1 h, spin-dried by a rotary evaporator, and the intermediate was obtained by column chromatography.

Add the intermediate (15mmol), CH ₃ OH-H ₂ O (V _CH3OH :V _H2O = 3:1) solution (15mL) to the reaction flask in turn, add LiOH·H ₂ O (45mmol) under ice-cooling, stir, TLC monitoring until the end of the reaction. Remove CH ₃ OH with a rotary evaporator, adjust the pH to about 3 with 4N HCl solution in an ice bath, a large amount of white solid precipitates, filter with suction, wash the filter cake twice with the filtrate and twice with acetone, and dry at 50°C to obtain intermediate.

Add the above-mentioned intermediate compound (1mmol), HBTU/TBTU (1.2mmol), DCM (3mL) in sequence in the reaction flask, add DIPEA3mmol, gatifloxacin (1mmol) under ice bath, stir under temperature control, TLC monitoring until the end of the reaction . Add 20 mL of DCM, transfer to a separatory funnel, wash with saturated Na ₂ CO ₃ aqueous solution (20 mL×1), 0.5N HCl solution (20 mL×1), saturated brine (20 mL×1), and dry with anhydrous Na ₂ SO ₄ for 1 h , spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 566.2409.

Example 26

Add glycine (50mmol) and methanol 100mL to the reaction bottle, slowly add SOCl ₂ (125mmol) dropwise under ice bath, after the drop is complete, transfer to a 60°C water bath to stir and reflux for reaction, monitor by TLC until the reaction is complete. The methanol and most of the SOCl ₂ were removed by rotary evaporation under reduced pressure, 20 mL of methanol was added, the rotary evaporation was repeated to remove as much SOCl ₂ as possible, and the corresponding glycine methyl ester hydrochloride was obtained by vacuum drying.

Add INA (20mmol), HOBt (24mmol), DCC (24mmol) and DCM (20mL) in turn to the reaction flask, stir for 10min, then add DIPEA under ice-cooling, add glycine methyl ester hydrochloride (22mmol) after 0.5-1h, TLC monitoring until the end of the reaction. Put it in the refrigerator for 2h, filter with suction, wash the filter cake with DCM, wash the filtrate with a saturated Na ₂ CO ₃ aqueous solution (20mL×2), extract the aqueous phase with DCM (30mL×3), combine the organic phases, anhydrous Na ₂ SO ₄ was dried for 1 h, spin-dried by a rotary evaporator, and the intermediate was obtained by column chromatography.

Add the intermediate (15mmol), CH ₃ OH-H ₂ O (V _CH3OH :V _H2O = 3:1) solution (15mL) to the reaction flask in turn, add LiOH·H ₂ O (45mmol) under ice-cooling, stir, TLC monitoring until the end of the reaction. Remove CH ₃ OH with a rotary evaporator, adjust the pH to about 3 with 4N HCl solution in an ice bath, a large amount of white solid precipitates, filter with suction, wash the filter cake twice with the filtrate and twice with acetone, and dry at 50°C to obtain intermediate.

Add the above-mentioned intermediate compound (1mmol), HBTU/TBTU (1.2mmol), DCM (3mL), Et ₃ N 3mmol, norfloxacin (1mmol) in sequence in the reaction flask, stir under temperature control, and monitor by TLC until the end of the reaction . Add 20 mL of DCM, transfer to a separatory funnel, wash with saturated Na ₂ CO ₃ aqueous solution (20 mL×1), 0.5N HCl solution (20 mL×1), saturated brine (20 mL×1), and dry over anhydrous Na ₂ SO ₄ After 1h, it was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 482.1853.

Example 27

Add alanine (50mmol) and methanol 100mL to the reaction flask, slowly add SOCl ₂ (125mmol) dropwise under ice bath, after the drop is complete, transfer to a 60°C water bath to stir and reflux for reaction, monitor by TLC until the reaction is complete. The methanol and most of the SOCl ₂ were removed by rotary evaporation under reduced pressure, 20 mL of methanol was added, the rotary evaporation was repeated to remove as much SOCl ₂ as possible, and the corresponding alanine methyl ester hydrochloride was obtained by vacuum drying.

Add INA (20mmol), HOBt (24mmol), DCC (24mmol), DCM (20mL) in turn to the reaction flask, stir for 10min, add DIPEA under ice bath, add alanine methyl ester hydrochloride (22mmol ), TLC monitoring until the end of the reaction. Put it in the refrigerator for 2h, filter with suction, wash the filter cake with DCM, wash the filtrate with a saturated Na ₂ CO ₃ aqueous solution (20mL×2), extract the aqueous phase with DCM (30mL×3), combine the organic phases, anhydrous Na ₂ SO ₄ was dried for 1 h, spin-dried by a rotary evaporator, and the intermediate was obtained by column chromatography.

Add the intermediate (15mmol), CH ₃ OH-H ₂ O (V _CH3OH :V _H2O = 3:1) solution (15mL) to the reaction flask in turn, add LiOH·H ₂ O (45mmol) under ice-cooling, stir, TLC monitoring until the end of the reaction. Remove CH ₃ OH with a rotary evaporator, adjust the pH to about 3 with 4N HCl solution in an ice bath, a large amount of white solid precipitates, filter with suction, wash the filter cake twice with the filtrate and twice with acetone, and dry at 50°C to obtain intermediate.

Add the above-mentioned intermediate compound (1mmol), HBTU/TBTU (1.2mmol), DCM (3mL) in sequence in the reaction flask, add DIPEA3mmol, norfloxacin (1mmol) under ice bath, stir under temperature control, TLC monitoring until the end of the reaction . Add 20 mL of DCM, transfer to a separatory funnel, wash with saturated Na ₂ CO ₃ aqueous solution (20 mL×1), 0.5N HCl solution (20 mL×1), saturated brine (20 mL×1), and dry with anhydrous Na ₂ SO ₄ for 1 h , spin-dried on a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 496.2015.

Example 28

Add 3-aminopropionic acid (50mmol) and methanol 100mL to the reaction flask, slowly add SOCl ₂ (125mmol) dropwise under ice bath, after the drop is complete, transfer to a 60°C water bath to stir and reflux for reaction, monitor by TLC until the reaction is complete. The methanol and most of the SOCl ₂ were removed by rotary evaporation under reduced pressure, 20 mL of methanol was added, the rotary evaporation was repeated to remove as much SOCl ₂ as possible, and the corresponding methyl 3-aminopropionate hydrochloride was obtained by vacuum drying.

Add INA (20mmol), HOBt (24mmol), DCC (24mmol), DCM (20mL) in turn into the reaction flask, stir for 10min, then add DIPEA under ice bath, add 3-alanine methyl ester hydrochloride after 0.5~1h (22mmol), TLC monitoring until the end of the reaction. Put it in the refrigerator for 2h, filter with suction, wash the filter cake with DCM, wash the filtrate with a saturated Na ₂ CO ₃ aqueous solution (20mL×2), extract the aqueous phase with DCM (30mL×3), combine the organic phases, anhydrous Na ₂ SO ₄ was dried for 1 h, spin-dried by a rotary evaporator, and the intermediate was obtained by column chromatography.

Add the intermediate (15mmol), CH ₃ OH-H ₂ O (V _CH3OH :V _H2O = 3:1) solution (15mL) to the reaction flask in turn, add LiOH·H ₂ O (45mmol) under ice-cooling, stir, TLC monitoring until the end of the reaction. Remove CH ₃ OH with a rotary evaporator, adjust the pH to about 3 with 4N HCl solution in an ice bath, a large amount of white solid precipitates, filter with suction, wash the filter cake twice with the filtrate and twice with acetone, and dry at 50°C to obtain intermediate.

Add the above-mentioned intermediate compound (1mmol), HBTU/TBTU (1.2mmol), DCM (3mL) in sequence in the reaction flask, add DIPEA3mmol, norfloxacin (1mmol) under ice bath, stir under temperature control, TLC monitoring until the end of the reaction . Add 20 mL of DCM, transfer to a separatory funnel, wash with saturated Na ₂ CO ₃ aqueous solution (20 mL×1), 0.5N HCl solution (20 mL×1), saturated brine (20 mL×1), and dry with anhydrous Na ₂ SO ₄ for 1 h , spin-dried on a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 496.2064.

Example 29

Add 4-aminobutyric acid (50mmol) and 100mL of methanol into the reaction bottle, slowly add SOCl ₂ (125mmol) dropwise under ice bath, after the drop is complete, transfer to a 60°C water bath to stir and reflux for reaction, monitor by TLC until the reaction is complete. The methanol and most of the SOCl ₂ were removed by rotary evaporation under reduced pressure, 20 mL of methanol was added, the rotary evaporation was repeated to remove as much SOCl ₂ as possible, and the corresponding 4-aminobutyric acid methyl ester hydrochloride was obtained by vacuum drying.

Add INA (20mmol), HOBt (24mmol), DCC (24mmol), DCM (20mL) in turn into the reaction flask, stir for 10min, add DIPEA under ice bath, add 4-aminobutyric acid methyl ester hydrochloride after 0.5~1h (22mmol), TLC monitoring until the end of the reaction. Put it in the refrigerator for 2 hours, filter with suction, wash the filter cake with DCM, wash the filtrate with saturated Na ₂ CO ₃ aqueous solution (20mL×2), extract the aqueous phase with DCM (30mL×3), combine the organic phases, and anhydrous Na ₂ SO ₄ was dried for 1 h, spin-dried by a rotary evaporator, and the intermediate was obtained by column chromatography.

Add the intermediate (15mmol), CH ₃ OH-H ₂ O (V _CH3OH :V _H2O = 3:1) solution (15mL) to the reaction flask in turn, add LiOH·H ₂ O (45mmol) under ice-cooling, stir, TLC monitoring until the end of the reaction. Remove CH ₃ OH with a rotary evaporator, adjust the pH to about 3 with 4N HCl solution in an ice bath, a large amount of white solid precipitates, filter with suction, wash the filter cake twice with the filtrate and twice with acetone, and dry at 50°C to obtain intermediate.

Add the above-mentioned intermediate compound (1mmol), HBTU/TBTU (1.2mmol), DCM (3mL) in sequence in the reaction flask, add DIPEA3mmol, norfloxacin (1mmol) under ice bath, stir under temperature control, TLC monitoring until the end of the reaction . Add 20 mL of DCM, transfer to a separatory funnel, wash with saturated Na ₂ CO ₃ aqueous solution (20 mL×1), 0.5N HCl solution (20 mL×1), saturated brine (20 mL×1), and dry with anhydrous Na ₂ SO ₄ for 1 h , spin-dried on a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 510.2148.

Example 30

Add glycine (50mmol) and methanol 100mL to the reaction bottle, slowly add SOCl ₂ (125mmol) dropwise under ice bath, after the drop is complete, transfer to a 60°C water bath to stir and reflux for reaction, monitor by TLC until the reaction is complete. The methanol and most of the SOCl ₂ were removed by rotary evaporation under reduced pressure, 20 mL of methanol was added, the rotary evaporation was repeated to remove as much SOCl ₂ as possible, and the corresponding glycine methyl ester hydrochloride was obtained by vacuum drying.

Add INA (20mmol), HOBt (24mmol), DCC (24mmol) and DCM (20mL) in turn to the reaction flask, stir for 10min, then add DIPEA under ice-cooling, add glycine methyl ester hydrochloride (22mmol) after 0.5-1h, TLC monitoring until the end of the reaction. Put it in the refrigerator for 2h, filter with suction, wash the filter cake with DCM, wash the filtrate with a saturated Na ₂ CO ₃ aqueous solution (20mL×2), extract the aqueous phase with DCM (30mL×3), combine the organic phases, anhydrous Na ₂ SO ₄ was dried for 1 h, spin-dried by a rotary evaporator, and the intermediate was obtained by column chromatography.

Add the intermediate (15mmol), CH ₃ OH-H ₂ O (V _CH3OH :V _H2O = 3:1) solution (15mL) to the reaction flask in turn, add LiOH·H ₂ O (45mmol) under ice-cooling, stir, TLC monitoring until the end of the reaction. Remove CH ₃ OH with a rotary evaporator, adjust the pH to about 3 with 4N HCl solution in an ice bath, a large amount of white solid precipitates, filter with suction, wash the filter cake twice with the filtrate and twice with acetone, and dry at 50°C to obtain intermediate.

Add the above-mentioned intermediate compound (1mmol), HBTU/TBTU (1.2mmol), DCM (3mL), Et ₃ N 3mmol, and enoxacin (1mmol) in sequence in the reaction flask, stir under temperature control, and monitor by TLC until the reaction is complete . Add 20 mL of DCM, transfer to a separatory funnel, wash with saturated Na ₂ CO ₃ aqueous solution (20 mL×1), 0.5N HCl solution (20 mL×1), saturated brine (20 mL×1), and dry over anhydrous Na ₂ SO ₄ After 1h, it was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 483.1754.

Example 31

Add alanine (50mmol) and methanol 100mL to the reaction flask, slowly add SOCl ₂ (125mmol) dropwise under ice bath, after the drop is complete, transfer to a 60°C water bath to stir and reflux for reaction, monitor by TLC until the reaction is complete. The methanol and most of the SOCl ₂ were removed by rotary evaporation under reduced pressure, 20 mL of methanol was added, the rotary evaporation was repeated to remove as much SOCl ₂ as possible, and the corresponding alanine methyl ester hydrochloride was obtained by vacuum drying.

Add INA (20mmol), HOBt (24mmol), DCC (24mmol), DCM (20mL) in turn to the reaction flask, stir for 10min, add DIPEA under ice bath, add alanine methyl ester hydrochloride (22mmol ), TLC monitoring until the end of the reaction. Put it in the refrigerator for 2h, filter with suction, wash the filter cake with DCM, wash the filtrate with a saturated Na ₂ CO ₃ aqueous solution (20mL×2), extract the aqueous phase with DCM (30mL×3), combine the organic phases, anhydrous Na ₂ SO ₄ was dried for 1 h, spin-dried by a rotary evaporator, and the intermediate was obtained by column chromatography.

Add the intermediate (15mmol), CH ₃ OH-H ₂ O (V _CH3OH :V _H2O = 3:1) solution (15mL) to the reaction flask in turn, add LiOH·H ₂ O (45mmol) under ice-cooling, stir, TLC monitoring until the end of the reaction. Remove CH ₃ OH with a rotary evaporator, adjust the pH to about 3 with 4N HCl solution in an ice bath, a large amount of white solid precipitates, filter with suction, wash the filter cake twice with the filtrate and twice with acetone, and dry at 50°C to obtain intermediate.

Add the above-mentioned intermediate compound (1mmol), HBTU/TBTU (1.2mmol), DCM (3mL) in turn to the reaction flask, add DIPEA3mmol, lomefloxacin (1mmol) under ice bath, stir under temperature control, and monitor by TLC until the reaction is complete. Add 20 mL of DCM, transfer to a separatory funnel, wash with saturated Na ₂ CO ₃ aqueous solution (20 mL×1), 0.5N HCl solution (20 mL×1), saturated brine (20 mL×1), and dry with anhydrous Na ₂ SO ₄ for 1 h , spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 528.2053.

Example 32

Add 5mmol of GAT, 15mL of DCM, and 7.5mmol of NaHCO ₃ into a 100mL round-bottomed flask in sequence, and after stirring evenly, slowly add a solution of 2.5mmol of solid phosgene in 5mL of dichloromethane (DCM) dropwise under an ice bath. After the dropwise addition, move to room temperature to stir the reaction, and monitor the reaction progress by TLC. After the reaction was completed, 10 mL of ice-cold saturated NaCl solution was added, and 2N HCl solution was added to adjust the pH value to 4-5, and the liquids were separated. Wash with 10 mL of saturated NaCl solution, separate the layers, dry the organic phase over anhydrous Na ₂ SO ₄ , and remove the solvent by rotary evaporation to obtain an intermediate.

Add 2mmol of piperidine, 3mL of CHCl ₃ , and 2mmol of K ₂ CO ₃ into a 100mL round bottom flask in sequence, and stir at room temperature for 30min. Add 1 mmol of intermediate, and move to 35°C water bath for reaction, TLC monitors the reaction progress. After the reaction, if the product is easily soluble in water, filter with suction, and wash the filter cake with a mixture of DCM and CH ₃ OH. The filtrate was adjusted to pH 7-8 with HCl-EA solution, and dried with Na ₂ SO ₄ ; if the water solubility of the product was low, add 15mL DCM, 10mL H ₂ O, 2N HCl solution to adjust the pH value to 7-8, and separate the liquids. Wash with 10 mL of saturated NaCl solution, separate the layers, and dry the organic phase over anhydrous Na ₂ SO ₄ ; remove the solvent by rotary evaporation to obtain a crude product, and column chromatography (DCM/CH ₃ OH) to obtain a pure product, [M+Na] ⁺ 509.2171.

Example 33

Add 5mmol of GAT, 15mL of DCM, and 7.5mmol of NaHCO ₃ into a 100mL round-bottomed flask in sequence, and after stirring evenly, slowly add a solution of 2.5mmol of solid phosgene in 5mL of dichloromethane (DCM) dropwise under an ice bath. After the dropwise addition, move to room temperature to stir the reaction, and monitor the reaction progress by TLC. After the reaction was completed, 10 mL of ice-cold saturated NaCl solution was added, and 2N HCl solution was added to adjust the pH value to 4-5, and the liquids were separated. Wash with 10 mL of saturated NaCl solution, separate the layers, dry the organic phase over anhydrous Na ₂ SO ₄ , and remove the solvent by rotary evaporation to obtain an intermediate.

Add 2mmol of 1-methylpiperazine, 3mL of CHCl ₃ and 2mmol of K ₂ CO ₃ to a 100mL round bottom flask in sequence, and stir at room temperature for 30min. Add 1 mmol of intermediate, and move to 35°C water bath for reaction, TLC monitors the reaction progress. After the reaction, if the product is easily soluble in water, filter with suction, and wash the filter cake with a mixture of DCM and CH ₃ OH. The filtrate was adjusted to pH 7-8 with HCl-EA solution, and dried with Na ₂ SO ₄ ; if the water solubility of the product was low, add 15mL DCM, 10mL H ₂ O, 2N HCl solution to adjust the pH value to 7-8, and separate the liquids. Wash with 10 mL of saturated NaCl solution, separate the layers, and dry the organic phase with anhydrous Na ₂ SO ₄ ; the solvent was removed by rotary evaporation to obtain a crude product, and column chromatography (DCM/CH ₃ OH) obtained a pure product, [M+H] ⁺ 502.2460.

Example 34

Add 5mmol of GAT, 15mL of DCM, and 7.5mmol of NaHCO ₃ into a 100mL round-bottomed flask in sequence, and after stirring evenly, slowly add 2.5mmol of solid phosgene in 5mL of DCM solution dropwise under an ice bath. After the dropwise addition, move to room temperature to stir the reaction, and monitor the reaction progress by TLC. After the reaction was completed, 10 mL of ice-cold saturated NaCl solution was added, and 2N HCl solution was added to adjust the pH value to 4-5, and the liquids were separated. Wash with 10 mL of saturated NaCl solution, separate the layers, dry the organic phase over anhydrous Na ₂ SO ₄ , and remove the solvent by rotary evaporation to obtain an intermediate.

Add 2 mmol of thiosemicarbazide, _{3 mL of CHCl 3} , and 2 mmol of K ₂ CO ₃ to a 100 mL round-bottom flask in sequence, and stir at room temperature for 30 min. Add 1 mmol of intermediate, and move to 35°C water bath for reaction, TLC monitors the reaction progress. After the reaction, if the product is easily soluble in water, filter with suction, and wash the filter cake with a mixture of DCM and CH ₃ OH. The filtrate was adjusted to pH 7-8 with HCl-EA solution, and dried with Na ₂ SO ₄ ; if the water solubility of the product was low, add 15mL DCM, 10mL H ₂ O, 2N HCl solution to adjust the pH value to 7-8, and separate the liquids. Wash with 10 mL of saturated NaCl solution, separate the layers, and dry the organic phase with anhydrous Na ₂ SO ₄ ; the solvent was removed by rotary evaporation to obtain a crude product, and column chromatography (DCM/CH ₃ OH) obtained a pure product, [M+H] ⁺ 493.1673.

Example 35

Add 5mmol of GAT, 15mL of DCM, and 7.5mmol of NaHCO ₃ into a 100mL round-bottomed flask in sequence, and after stirring evenly, slowly add 2.5mmol of solid phosgene in 5mL of DCM solution dropwise under an ice bath. After the dropwise addition, move to room temperature to stir the reaction, and monitor the reaction progress by TLC. After the reaction was completed, 10 mL of ice-cold saturated NaCl solution was added, and 2N HCl solution was added to adjust the pH value to 4-5, and the liquids were separated. Wash with 10 mL of saturated NaCl solution, separate the layers, dry the organic phase over anhydrous Na ₂ SO ₄ , and remove the solvent by rotary evaporation to obtain an intermediate.

Add 2mmol of 4-benzylpiperazine, 3mL of CHCl ₃ , and 2mmol of K ₂ CO ₃ to a 100mL round bottom flask in sequence, and stir at room temperature for 30min. Add 1 mmol of intermediate, and move to 35°C water bath for reaction, TLC monitors the reaction progress. After the reaction, if the product is easily soluble in water, filter with suction, and wash the filter cake with a mixture of DCM and CH ₃ OH. The filtrate was adjusted to pH 7-8 with HCl-EA solution, and dried with Na ₂ SO ₄ ; if the water solubility of the product was low, add 15mL DCM, 10mL H ₂ O, 2N HCl solution to adjust the pH value to 7-8, and separate the liquids. Wash with 10 mL of saturated NaCl solution, separate the layers, and dry the organic phase over anhydrous Na ₂ SO ₄ ; the solvent was removed by rotary evaporation to obtain a crude product, and column chromatography (DCM/CH ₃ OH) obtained a pure product, [M+H] ⁺ 578.2773.

Example 36

Add 2-aminoethanol 2mmol, CHCl ₃ 3mL, K ₂ CO ₃ 2mmol in sequence to a 100mL round bottom flask, and stir at room temperature for 30min. Add 1 mmol of the compound of Example 1, and move to 35° C. water bath for reaction, and monitor the reaction progress by TLC. After the reaction, 15 mL of DCM, 10 mL of H ₂ O, and 1N HCl solution were added to adjust the pH value to 7-8, and the liquids were separated. Wash with 10 mL of saturated NaCl solution, separate the layers, dry the organic phase over anhydrous Na ₂ SO ₄ , and spin evaporate to obtain a crude product, and column chromatography to obtain a pure product, [M+H] ⁺ 477.2144.

Example 37

To a 100mL round bottom flask, add 2mmol, CHCl ₃ 3mL, K ₂ CO ₃ 2mmol, stirred at room temperature for 30min. Add 1 mmol of the compound of Example 1, and move to 35° C. water bath for reaction, and monitor the reaction progress by TLC. After the reaction, 15 mL of DCM, 10 mL of H ₂ O, and 1N HCl solution were added to adjust the pH value to 7-8, and the liquids were separated. Wash with 10 mL of saturated NaCl solution, separate the layers, dry the organic phase over anhydrous Na ₂ SO ₄ , and spin evaporate to obtain a crude product, and column chromatography to obtain a pure product, [M+H] ⁺ 521.2406.

Example 38

Add 2-(methylamino)ethyl-1-ol 2mmol, CHCl ₃ 3mL, K ₂ CO ₃ 2mmol in sequence to a 100mL round bottom flask, and stir at room temperature for 30min. Add 1 mmol of the compound of Example 1, and move to 35° C. water bath for reaction, and monitor the reaction progress by TLC. After the reaction, 15 mL of DCM, 10 mL of H ₂ O, and 1N HCl solution were added to adjust the pH value to 7-8, and the liquids were separated. Wash with 10 mL of saturated NaCl solution, separate the layers, dry the organic phase over anhydrous Na ₂ SO ₄ , and spin evaporate to obtain a crude product, and column chromatography to obtain a pure product, [M+H] ⁺ 491.2300.

Example 39

To a 100mL round bottom flask, add 2mmol, CHCl ₃ 3mL, NaHCO ₃ 5mmol, stirred at room temperature for 30min. Add 1 mmol of the compound of Example 2, and move it to a 50°C water bath for reaction, and monitor the progress of the reaction by TLC. After the reaction, 15 mL of DCM, 10 mL of H ₂ O, and 2N HCl solution were added to adjust the pH value to 7-8, and the liquids were separated. Washed with 10 mL of saturated NaCl solution, separated, dried over anhydrous Na ₂ SO ₄ , and rotary evaporated to obtain a crude product, which was purified by column chromatography, [M+H] ⁺ 535.2563.

Example 40

1-Methylpiperazine 2mmol, CHCl ₃ 3mL, NaHCO ₃ 5mmol were sequentially added into a 100mL round bottom flask, and stirred at room temperature for 30min. Add 1 mmol of the compound of Example 2, and move it to a 50°C water bath for reaction, and monitor the progress of the reaction by TLC. After the reaction, 15 mL of DCM, 10 mL of H ₂ O, and 2N HCl solution were added to adjust the pH value to 7-8, and the liquids were separated. Washed with 10 mL of saturated NaCl solution, separated, dried over anhydrous Na ₂ SO ₄ , and rotary evaporated to obtain a crude product, which was purified by column chromatography, [M+H] ⁺ 530.2773.

Example 41

Into a 100 mL round bottom flask, 2 mmol of piperidine, _{3 mL of CHCl 3} , and 5 mmol of NaHCO ₃ were sequentially added, and stirred at room temperature for 30 min. Add 1 mmol of the compound of Example 2, and move it to a 50°C water bath for reaction, and monitor the progress of the reaction by TLC. After the reaction, 15 mL of DCM, 10 mL of H ₂ O, and 2N HCl solution were added to adjust the pH value to 7-8, and the liquids were separated. Wash with 10 mL of saturated NaCl solution, separate the layers, dry over anhydrous Na ₂ SO ₄ , and rotary evaporate to obtain a crude product, and column chromatography to obtain a pure product, [M+H] ⁺ 543.2613.

Example 42

2mmol of 4-benzylpiperidine, 3mL of CHCl ₃ , and 5mmol of NaHCO ₃ were sequentially added into a 100mL round bottom flask, and stirred at room temperature for 30min. Add 1 mmol of the compound of Example 2, and move it to a 50°C water bath for reaction, and monitor the progress of the reaction by TLC. After the reaction, 15 mL of DCM, 10 mL of H ₂ O, and 2N HCl solution were added to adjust the pH value to 7-8, and the liquids were separated. Washed with 10 mL of saturated NaCl solution, separated, dried over anhydrous Na ₂ SO ₄ , and rotary evaporated to obtain a crude product, which was purified by column chromatography, [M+H] ⁺ 634.3035.

Example 43

Add 3mmol of 1H-1,2,4-triazole and 2mL of DMF to a 100mL round bottom flask successively, stir at room temperature, add 4mmol of NaH, stir at room temperature for 30min, add 1mmol of the compound of Example 2, and transfer to 80°C water bath for reaction. The progress of the reaction was monitored by TLC. After the reaction, 20 mL of ice-cold saturated NaCl solution was added, and 2N HCl solution was added to adjust the pH value to 5-6, and a large amount of solids precipitated out. After suction filtration, the filter cake was washed with ice-cold saturated NaCl solution, washed once with ice water, and dried to obtain a crude product. The pure product was obtained by column chromatography and thin layer chromatography, [M+Na] ⁺ 521.19192.

Example 44

Add 3mmol of 1H-1,2,4-triazol-3-amine and 2mL of DMF to a 100mL round bottom flask successively, stir at room temperature, add 4mmol of NaH, stir at room temperature for 30min, add 1mmol of the compound of Example 2, and transfer to 80 ℃ water bath reaction. The progress of the reaction was monitored by TLC. After the reaction, 20 mL of ice-cold saturated NaCl solution was added, and 2N HCl solution was added to adjust the pH value to 5-6, and a large amount of solids precipitated out. After suction filtration, the filter cake was washed with ice-cold saturated NaCl solution, washed once with ice water, and dried to obtain a crude product. The pure product was obtained by column chromatography and thin layer chromatography, [M+Na] ⁺ 536.20282.

Example 45

Add 3mmol of 1H-tetrazolium-5-amine and 2mL of DMF to a 100mL round bottom flask, stir at room temperature, add 4mmol of NaH, stir for 30min at room temperature, add 1mmol of the compound of Example 2, and transfer to 80°C water bath for reaction. The progress of the reaction was monitored by TLC. After the reaction, 20 mL of ice-cold saturated NaCl solution was added, and 2N HCl solution was added to adjust the pH value to 5-6, and a large amount of solids precipitated out. After suction filtration, the filter cake was washed with ice-cold saturated NaCl solution, washed once with ice water, and dried to obtain a crude product, which was purified by column chromatography and thin-layer chromatography, [M+H] ⁺ 515.2135.

Example 46

Add 3mmol of 5-(difluoromethoxy)-1H-benzo[d]imidazole-2-thiol and 2mL of DMF in sequence to a 100mL round bottom flask, stir at room temperature, add 4mmol of NaH, stir at room temperature for 30min, add Example 2 compound 1mmol, and moved to 80 ℃ water bath reaction. The progress of the reaction was monitored by TLC. After the reaction, 20 mL of ice-cold saturated NaCl solution was added, and 2N HCl solution was added to adjust the pH value to 5-6, and a large amount of solids precipitated out. After suction filtration, the filter cake was washed with ice-cold saturated NaCl solution, washed once with ice water, and dried to obtain a crude product. The pure product was obtained by column chromatography and thin layer chromatography, [M+H] ⁺ 646.1942.

Example 47

Add 3mmol of benzo[d]thiazole-2-thiol and 2mL of DMF to a 100mL round bottom flask successively, stir at room temperature, add 4mmol of NaH, stir at room temperature for 30min, add 1mmol of the compound of Example 2, and transfer to 80°C water bath for reaction. The progress of the reaction was monitored by TLC. After the reaction, 20 mL of ice-cold saturated NaCl solution was added, and 2N HCl solution was added to adjust the pH value to 5-6, and a large amount of solids precipitated out. After suction filtration, the filter cake was washed with ice-cold saturated NaCl solution, washed once with ice water, and dried to obtain a crude product. The pure product was obtained by column chromatography and thin layer chromatography, [M+H] ⁺ 597.1636.

Example 48

Add 3mmol of 5-methyl-1,3,4-thiadiazole-2-thiol and 2mL of DMF to a 100mL round bottom flask in turn, stir at room temperature, add 4mmol of NaH, stir at room temperature for 30min, add 1mmol of the compound of Example 2, And moved to 80 ℃ water bath reaction. The progress of the reaction was monitored by TLC. After the reaction, 20 mL of ice-cold saturated NaCl solution was added, and 2N HCl solution was added to adjust the pH value to 5-6, and a large amount of solids precipitated out. After suction filtration, the filter cake was washed with ice-cold saturated NaCl solution, washed once with ice water, and dried to obtain a crude product. The pure product was obtained by column chromatography and thin layer chromatography, [M+Na] ⁺ 584.1408.

Example 49

0.204 g (3 mmol) of 1H-imidazole, 10 mL of toluene, and 0.484 g (1 mmol) of the compound of Example 3 were added to a 100 mL round bottom flask. The reaction was carried out under temperature control, and the progress of the reaction was detected by TLC. After the reaction was completed, the solvent toluene was removed by rotary evaporation. The pure product was obtained by solid column chromatography and thin layer chromatography (DCM:CH ₃ OH=30:1), [M+H] ⁺ 512.2304.

Example 50

3 mmol of 2-methyl-1H-imidazole, 10 mL of toluene, and 0.484 g (1 mmol) of the compound of Example 3 were added to a 100 mL round bottom flask. The reaction was carried out under temperature control, and the progress of the reaction was detected by TLC. After the reaction was completed, the solvent toluene was removed by rotary evaporation, and the pure product was obtained by solid column chromatography and thin layer chromatography (DCM:CH ₃ OH=30:1), [M+H] ⁺ 526.2460.

Example 51

3 mmol of 1H-pyrazole, 10 mL of toluene, and 0.484 g (1 mmol) of the compound of Example 3 were added to a 100 mL round bottom flask. The reaction was carried out under temperature control, and the progress of the reaction was detected by TLC. After the reaction was completed, the solvent toluene was removed by rotary evaporation, and the pure product was obtained by solid column chromatography and thin layer chromatography (DCM:CH ₃ OH=30:1), [M+Na] ⁺ 534.2123.

Example 52

3,5-Dimethyl-1H-pyrazole 3mmol, toluene 10mL, and Example 3 compound 0.484g (1mmol) were added to a 100mL round bottom flask. The reaction was carried out under temperature control, and the progress of the reaction was detected by TLC. After the reaction was completed, the solvent toluene was removed by rotary evaporation, and the pure product was obtained by solid column chromatography and thin layer chromatography (DCM:CH ₃ OH=30:1), [M+H] ⁺ 540.2617.

Example 53

Into a 100 mL round bottom flask were added 3 mmol of 1H-benzo[d]imidazole, 10 mL of toluene, and 0.484 g (1 mmol) of the compound of Example 5. Temperature control reaction, TLC detection reaction progress. After the reaction was completed, the solvent toluene was removed by rotary evaporation, and the pure product was obtained by solid column chromatography and thin layer chromatography (DCM:CH ₃ OH=30:1), [M+H] ⁺ 562.2460.

Example 54

3 mmol of 1H-1,2,3-triazole, 10 mL of toluene, and 0.484 g (1 mmol) of the compound of Example 5 were added to a 100 mL round bottom flask. The reaction was carried out under temperature control, and the progress of the reaction was detected by TLC. After the reaction was completed, the solvent toluene was removed by rotary evaporation. The pure product was obtained by solid column chromatography and thin layer chromatography (DCM:CH ₃ OH=30:1), [M+Na]+535.2076.

Example 55

3 mmol of 1H-tetrazole, 10 mL of toluene, and 0.484 g (1 mmol) of the compound of Example 5 were added to a 100 mL round bottom flask. The reaction was carried out under temperature control, and the progress of the reaction was detected by TLC. After the reaction was completed, the solvent toluene was removed by rotary evaporation, and the pure product was obtained by solid column chromatography and thin layer chromatography (DCM:CH ₃ OH=30:1), [M+Na] ⁺ 536.2028.

Example 56

3 mmol of 5-methyl-1H-tetrazole, 10 mL of toluene, and 0.484 g (1 mmol) of the compound of Example 5 were added to a 100 mL round bottom flask. The reaction was carried out under temperature control, and the progress of the reaction was detected by TLC. After the reaction was completed, the solvent toluene was removed by rotary evaporation, and the pure product was obtained by solid column chromatography and thin layer chromatography (DCM:CH ₃ OH=30:1), [M+Na] ⁺ 550.2185.

Example 57

3 mmol of 5-amino-1H-tetrazole, 10 mL of toluene, and 0.484 g (1 mmol) of the compound of Example 5 were added to a 100 mL round bottom flask. The reaction was carried out under temperature control, and the progress of the reaction was detected by TLC. After the reaction was completed, the solvent toluene was removed by rotary evaporation, and the pure product was obtained by solid column chromatography and thin layer chromatography (DCM:CH ₃ OH=30:1), [M+Na] ⁺ 551.2137.

Example 58

Into a 100 mL round bottom flask were added 3 mmol of 5-(difluoromethoxy)-1H-benzo[d]imidazole-2-thiol, 10 mL of toluene, and 0.484 g (1 mmol) of the compound of Example 5. The reaction was carried out under temperature control, and the progress of the reaction was detected by TLC. After the reaction was completed, the solvent toluene was removed by rotary evaporation, and the pure product was obtained by solid column chromatography and thin layer chromatography (DCM:CH ₃ OH=30:1), [M+Na] ⁺ 682.1918.

Example 59

Into a 100 mL round bottom flask were added 3 mmol of 1-methyl-1H-tetrazole-5-thiol, 10 mL of toluene, and 0.484 g (1 mmol) of the compound of Example 5. The reaction was carried out under temperature control, and the progress of the reaction was detected by TLC. After the reaction was completed, the solvent toluene was removed by rotary evaporation, and the pure product was obtained by solid column chromatography and thin layer chromatography (DCM:CH ₃ OH=30:1), [M+Na] ⁺ 582.1905.

Example 60

A 100 mL round bottom flask was charged with 3 mmol of benzo[d]thiazole-2-thiol, 10 mL of toluene, and 0.484 g (1 mmol) of the compound of Example 5. Temperature control reaction, TLC detection reaction progress. After the reaction was completed, the solvent toluene was removed by rotary evaporation, and the pure product was obtained by solid column chromatography and thin layer chromatography (DCM:CH ₃ OH=30:1), [M+Na] ⁺ 633.1612.

Example 61

3 mmol of 5-amino-1,3,4-thiadiazole-2-thiol, 10 mL of toluene, and 0.484 g (1 mmol) of the compound of Example 5 were added to a 100 mL round bottom flask. Temperature control reaction, TLC detection reaction progress. After the reaction was completed, the solvent toluene was removed by rotary evaporation. The pure product was obtained by solid column chromatography and thin layer chromatography (DCM:CH ₃ OH=30:1), [M+H] ⁺ 577.1698.

Example 62

Add 1mmol Clinfloxacin and 2mL DCM to a 100mL round bottom flask, cool in an ice bath, stir magnetically, add 3mmolNaHCO ₃ , after 20min, add 2.5mmol acetyl chloride solution in DCM (2mL) dropwise with a constant pressure dropping funnel (the dropping rate is about 1d/2s), the reaction was continued in an ice bath after dripping, followed by TLC monitoring until the end of the reaction. Stop stirring, add H ₂ O 15mL and DCM 20mL, adjust the pH=3-4 with 1N HCl solution under stirring, separate the layers, extract the aqueous phase with DCM (15mL×1), combine the organic phases, wash with saturated NaCl solution (15mL×1 1), the organic phase was collected and dried over anhydrous Na ₂ SO ₄ . The crude product was spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, and dried in vacuum to obtain the product, [M+H] ⁺ 409.1119.

Example 63

Add CF ₃ COOH (1.2mmol), HBTU/TBTU (1.44mmol), DCM (3mL), Et ₃ N 3mmol, and Clinfloxacin (1mmol) sequentially into the reaction flask, stir under temperature control, and monitor by TLC until the reaction is complete. Add 20 mL of DCM, transfer to a separatory funnel, wash with saturated Na ₂ CO ₃ aqueous solution (20 mL×1), 0.5N HCl solution (20 mL×1), saturated brine (20 mL×1), and dry over anhydrous Na ₂ SO ₄ , spin-dried by a rotary evaporator, and the pure product was obtained by column chromatography, [M+H] ⁺ 462.0845.

Determination of Antibacterial Activity of Xerosis citrus

Determination method: Weigh 1 mg of sample and dissolve it in 50 μL of DMSO, and dilute to 550 μL with ultrapure water as sample mother solution (1.82 mg/mL). Take 10 μL of mother liquor in 1 mL of ultrapure water (0.02% Tween) as sample solution a (0.0182 mg/mL), and then prepare sample solutions b (0.0091 mg/mL) and c (0.00455 mg/mL) sequentially by doubling dilution method ), d (0.002275 mg/mL), e (0.0011375 mg/mL), f (0.00056875 mg/mL).

Canker sore bacteria that have been cultured on PDA medium for 3 days were washed with 5 mL LB liquid medium, added to 195 mL LB liquid medium, oscillated and mixed for later use. Add 450 μL of X. citrus bacteria liquid and 50 μL of the above-mentioned sample solutions of different concentrations (a to f) into each 2 mL centrifuge tube, so that the final concentrations of the samples in each mixed bacterial liquid are A (0.00182 mg/mL), B ( 0.00091mg/mL), C (0.000455mg/mL), D (0.0002275mg/mL), E (0.00011375mg/mL), F (0.000056875mg/mL), 28 ℃, 200r min ^-1 constant temperature shaking culture for 14h Then measure the OD value of each mixed bacterial solution at OD ₆₀₀ and calculate the inhibition rate (inhibition rate%=(OD _blank -OD _sample )/OD _blank ×100%). Each treatment was replicated three times.

The inhibitory effect of table 1 compound on citrus canker bacterium

The PBT data processing system was used to statistically analyze the test results of each agent, and the inhibitory rate, toxicity regression equation, EC ₅₀ , EC ₉₀ and correlation coefficient (r) of each agent against X. citri were obtained.

The inhibitory effect of table 2 compound on citrus canker bacterium

Determination of antibacterial activity of Citrus brown spot fungus

Determination method: wash the conidia of brown spot fungus that has been cultured on PDA medium for 7 days with 0.05% Tween 80, and filter it with four layers of sterile lens-cleaning paper for later use; weigh 1 mg of the compound sample and dissolve it in 50 μL DMSO, Dilute to 550 μL with ultrapure water as sample mother solution (1.82 mg/mL). Take 2 μL of the mother solution in 1 mL of PDA (0.02% Tween) as sample solution a (0.00364 mg/mL), and sequentially prepare sample solution b (0.00182 mg/mL) by doubling dilution method. And add 0.5mL sample solution and 1.5mLPDA culture medium in each well of 48-well plate, after making test final concentration be A (0.000910mg/mL) and B (0.000455mg/mL) respectively, inoculate 2 μ L conidia suspension, The colony diameter was measured after 3 days of light incubation at 28°C. Taking the colony inoculated on the PDA medium without sample solution as the blank control, the inhibition rate of different samples to pathogenic fungi was calculated (inhibition rate%=(colony diameter _blank -colony diameter _sample )/colony diameter _blank ×100%).

Table 3 Compound anti-citrus brown spot fungus activity result

As can be seen from the above table, even at relatively low concentrations (0.00182mg/mL), the compounds of the embodiments of the present invention still show excellent antibacterial activity to X. citri, while the compounds of the present invention also have a certain brown The antibacterial activity of spot fungus plays a certain role in the prevention and treatment of citrus brown spot while preventing and treating citrus canker.

Finally, it is noted that the above embodiments are only used to illustrate the technical solutions of the present invention without limitation. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be carried out Modifications or equivalent replacements without departing from the spirit and scope of the technical solution of the present invention shall be covered by the claims of the present invention.

Claims (10)

1. compound, its raceme, stereoisomer, dynamic isomer, nitrogen oxides or their pharmacy can connect shown in formula I By salt：

Wherein X is selected from：C1-C6 alkyl；C3-C6 cycloalkyl；Substituted or unsubstituted C6-C10 aryl, taking on the aryl Dai Jiwei is one or more, is independently selected from：Halogen；Amino；Hydroxyl；C1-C6 alkyl；C3-C6 cycloalkyl；

Z is selected from：N or C-R₇；R₇Selected from H；C1-C6 alkoxy or halogens；

R₁And R₂It is each independently selected from：H；C1-C6 alkyl；C1-C6 haloalkyls；C1-C6 alkoxies；Halogen；Hydroxyl；Amino or Cyano group；

Q₁Selected from H；Halogen；

R₃Or R₄It is each independently selected from H；C1-C6 alkyl, the C1-C6 alkyl can be optionally by hydroxyls；Amino；Halogen；Cyano group；Urea Base；Ghiourea group substitutes；

Or R₃, R₄5-8 circle heterocycles are collectively forming with N, the heterocycle optionally can be substituted by one or more following substituents：

Amino；Hydroxyl；Tertbutyloxycarbonyl；Benzyl or C1-C6 alkyl, the C1-C6 alkyl can be optionally by hydroxyls；Amino；Halogen； C1-C4 alkoxies substitute；

A₁, A₂, A₃, A₄It is independently selected from C-R₅Or N, and A₁, A₂In at least one be N；

R₅Selected from H；C1-C4 alkyl；C1-C4 alkoxies；Amino；Acylamino-；Hydroxyl；Halogen；Phenyl or benzyl；

R₆For 1-4, H is independently selected from, C1-C6 alkyl, the C1-C6 alkyl can be optionally by hydroxyl；Amino；Halogen or cyano group take Generation；C1-C6 alkoxies, the C1-C6 alkoxies can be optionally optionally substituted by halogen；

B₁Selected from S or NH；

Het is the 5-7 member hetero-aromatic rings containing at least one nitrogen-atoms；

R₈Selected from H, C1-C6 alkyl；

N, k is respectively 0 to 3 integer.

2. compound, its raceme, stereoisomer, dynamic isomer, nitrogen oxides or their pharmacy can as shown in Formula II Receive salt：

Wherein X is selected from：C1-C6 alkyl；C3-C6 cycloalkyl；Substituted or unsubstituted C6-C10 aryl, taking on the aryl Dai Jiwei is one or more, is independently selected from：Halogen；Amino；Hydroxyl；C1-C6 alkyl；C3-C6 cycloalkyl；

Z is selected from：N or C-R₇；R₇Selected from H；C1-C6 alkoxy or halogens；

R₁And R₂It is each independently selected from：H；C1-C6 alkyl；C1-C6 haloalkyls；C1-C6 alkoxies；Halogen；Hydroxyl；Amino or Cyano group；

R₈Selected from H, C1-C6 alkyl；

Q₂It is selected fromOr halogen or H；

Het is the 5-7 member hetero-aromatic rings containing at least one nitrogen-atoms；

N, k is respectively 0 to 3 integer；

M is 1 or 2.

3. compound as claimed in claim 1 or 2, its raceme, stereoisomer, dynamic isomer, nitrogen oxides or they Pharmaceutically acceptable salt be characterized in that：

The het is selected from

The 5-8 circle heterocycles are selected from：

4. the compound of following structure, its raceme, stereoisomer, dynamic isomer, nitrogen oxides or their pharmacy can Receive salt：

5. a kind of pharmaceutical composition, it is characterised in that including compound, its raceme, alloisomerism described in claim 1-4 Body, dynamic isomer, nitrogen oxides or their pharmaceutically acceptable salt.

A kind of 6. pharmaceutical preparation, it is characterised in that including compound described in claim 1-4, its raceme, stereoisomer, Dynamic isomer, nitrogen oxides or their pharmaceutically acceptable salt, and pharmaceutically acceptable carrier and/or auxiliary agent.

A kind of 7. compound medicine, it is characterised in that including compound described in claim 1-4, its raceme, stereoisomer, Dynamic isomer, nitrogen oxides or their pharmaceutically acceptable salt and other active components.

8. compound, its raceme, stereoisomer, dynamic isomer, nitrogen oxides or them as described in claim 1-4 Purposes of the pharmaceutically acceptable salt in the medicine for preparing preventing and treating citrus correlation disease.

9. purposes as claimed in claim 8, it is characterised in that the citrus correlation disease is citrus bacterial canker disease or citrus foxiness Disease.

A kind of 10. method for treating citrus correlation disease, it is characterised in that contain including being applied to citrus fruit tree and/or fruit Compound, its raceme, stereoisomer, dynamic isomer, nitrogen oxides or their pharmacy can as described in claim 1-4 Receive the medicament of salt.