Isoindolinone derivatives, processes for their preparation and use thereof

Abstract

A isoindolinone derivative is prepared by substituted or non-substituted isoindolinone, amino acid, biological amine and the like, and is a series of compounds with the structure shown in formula I, or formula II, or formula III, or formula IV. The novel isoindolinone derivative provided by the application takes an amide bond as a core structure, has superior prevention and treatment effect on various plant diseases, shows good biocompatibility and stability, and can promote the autophagy physiological phenomenon of biological cells. The innovative mechanism creates a new idea and method for the application of isoindolinone structure unit compounds in the agricultural field. The preparation method is relatively simple, raw materials are easy to obtain, cost is low, industrial production is easy, yield is high, economy is good, and the isoindolinone derivative has great popularization and application value.

Description

Technical Field

[0001] This invention relates to an isoindoline ketone derivative, its preparation method, and its application, belonging to the technical field of preparation and application of organic agricultural bactericides. Background Technology

[0002] Isoindolineone skeletons are widely found in natural products and are a class of nitrogen-containing heterocyclic compounds with important biological activities, which have been widely used in the fields of medicine and agriculture.

[0003] In the pharmaceutical field, isoindolone derivatives have anticholinesterase and β-amyloid aggregation activities, thus they can be used to improve cognitive function in Alzheimer's patients (Guzior et al., Bioorg MedChem. 2015, 23(7): 1629-1637; Hassanzadeh et al., Res Pharm Sci 2021, 16(5): 482-492); due to their tumor-associated carbonic anhydrase IX inhibitory activity, they exhibit antitumor effects and reduce cancer cell spread and metastasis (Abdel-Aziz et al., Bioorg Chem. 2018, 80: 706-713); in addition, isoindolone derivatives also have anti-ovalbumin-induced allergic asthma effects (Huang et al., Biomol Ther (Seoul). 2018, 26(6): 539-545), and anti-inflammatory effects by regulating cytokine production in spleen cell populations (Matalka et al.). al.,IntImmunopharmacol.2012,14(3):296-301), as well as antioxidant, anticancer, antiepileptic, antiviral, antibacterial and other biological activities; studies have shown that phthalimide derivatives in isoindolineone skeleton compounds exhibit better biological activity.

[0004] In the agricultural field, compounds containing isoindolinone structural units exhibit inhibitory activity against 4-hydroxyphenylpyruvate dioxygenase (HPPD) or protoporphyrinogen oxidase (PPO), showing potential herbicide applications (Huang et al., JAgric Food Chem. 2005, 53(20): 7908-14; 2009, 57(20): 9585-92; Jiang et al., JAgric Food Chem. 2011, 59(11): 6172-9; He et al., J Agric Food Chem 2019, 67(39): 10844-52). Among them, captan and folpet are derivatives containing isoindolinone skeletons, belonging to the trichloromethylthio (-SCCl3) class of non-systemic fungicides, which can effectively control fungal diseases of fruits, vegetables, rice and other crops (Abbott). It was first discovered in 1951 and has been used commercially for over 60 years. ( et al., Plant Dis. 2018, 102: 231-6).

[0005] However, many studies have reported that existing isoindolinone-based fungicides have genotoxicity, mutagenicity, and ecological risks due to their chemical reactions with thiols (Bridges, Mutat Res. 1975, 32(1): 3-34; Arce et al., Crit Rev Toxicol. 2010, 40(6): 546-74). For these reasons, there are currently almost no innovative reports on the application of isoindolinone structural units as fungicides in the agricultural field, both domestically and internationally. Summary of the Invention

[0006] To overcome the shortcomings of the prior art, this invention provides an isoindolinone derivative, its preparation method, and its application, with the aim of:

[0007] By organically combining isoindolone structural units with amino acids and biogenic amines, a series of novel isoindolone derivatives exhibiting good biocompatibility and promoting autophagy in biological cells were prepared for the prevention and control of various plant diseases. This innovative application of isoindolone structural unit compounds as fungicides in the agricultural field was also demonstrated.

[0008] To achieve the above objectives, the present invention first provides an isoindolinone derivative.

[0009] An isoindolinone derivative, which is a compound having a structure as shown in Formula I, or Formula II, or Formula III, or Formula IV:

[0010]

[0011] in:

[0012] R 1 It is any one of hydrogen, fluorine, chlorine, nitro, methoxy, methyl, or amino, or R 1 For substituted and unsubstituted phenyl groups, or R 1 It can be a halogenated or non-halogenated five- or six-membered heterocyclic group containing oxygen, nitrogen, and sulfur;

[0013] R 2 For C1~C 10 saturated alkyl groups, or R 2 For substituted or unsubstituted phenyl groups, or R 2 It is a five- or six-membered heterocyclic group containing oxygen, sulfur, and nitrogen, or R 2 for Group and R therein a It is any one of halophenyl, benzyl, pyridyl, furanyl, pyrimidinyl or five-membered cycloyl group;

[0014] R 3 It is a thiol group, or R 3 -(CH2) n NH2- group and n is 1 to 5, or R 3 for Group and R therein b It is sulfonyl or trifluoromethyl;

[0015] R c It is methyl or halophenyl.

[0016] Preferred:

[0017] The R 1 In substituted and unsubstituted phenyl groups, the substituted phenyl group is a halophenyl or Alternatively, it could be a trifluoromethyl-substituted phenyl group.

[0018] Preferably, the R 1 In the above, the halophenyl group is...

[0019] Preferred:

[0020] The R 1 It is a halogenated or non-halogenated five- or six-membered heterocyclic group containing oxygen, nitrogen, and sulfur, which is a halogenated or non-halogenated pyridinyl, pyrimidinyl, or furanyl group.

[0021] Preferred:

[0022] The R 1 In pyridinyl, pyrimidinyl, or furanyl groups that are halogenated or non-halogenated, the halogenated pyridinyl group is: Its halopyrimidine group is

[0023] Preferred:

[0024] The R 2 For C1~C 10 The saturated alkyl group is -CH2- or -(CH2)2- or -(CH2). 10 -;

[0025] The R 2 In substituted or unsubstituted phenyl groups, the substituted phenyl group is... Or for Or perhaps

[0026] The R 2 Among five- and six-membered heterocyclic groups containing oxygen, sulfur, and nitrogen, the nitrogen-containing five-membered heterocyclic group is: Its nitrogen-containing six-membered heterocyclic group is

[0027] Preferred:

[0028] The R 3 -(CH2) n The NH2- group is either -CH2NH2- or -(CH2)2NH2-.

[0029] Secondly, the present invention also provides a method for preparing the above-mentioned isoindoline ketone derivative.

[0030] The method for preparing the isoindolinone derivative includes steps a~b~c, or steps a~b~d, or steps a~b~d~e, or steps a~g~h in the following synthetic route, wherein step f is included before step g;

[0031] Synthesis route:

[0032]

[0033] in:

[0034] Step a involves reacting substituted or unsubstituted phthalic anhydride with an amino acid to obtain an aminolysis product;

[0035] Step b involves reacting the amino hydrolysate with thionyl chloride under reflux to form an acylated amino hydrolysate, i.e., intermediate A.

[0036] Step c involves reacting the ammonium chloride hydrolysate, intermediate A, with putrescine to obtain the isoindolinone derivative having the structure shown in Formula I.

[0037] Step d involves reacting the amyl chloride hydrolysate, i.e., intermediate A, with cystamine to obtain the isoindoline ketone derivative having the structure shown in Formula II;

[0038] Step e involves the isoindoline ketone derivative having the structure shown in Formula II undergoing a first group derivatization reaction to obtain the isoindoline ketone derivative having the structure shown in Formula III.

[0039] Step g involves the catalytic reaction of the amino hydrolysate with the tert-butylcarbonyl-protected putrescine obtained in step f (i.e., the amino protection reaction) to obtain intermediate B.

[0040] Step h involves a second group derivatization reaction of intermediate B to obtain the isoindoline ketone derivative having the structure shown in Formula IV.

[0041] Furthermore, step a includes:

[0042] Amine hydrolysis is carried out in glacial acetic acid with substituted or unsubstituted phthalic anhydride and an equimolar amount of amino acid. After the reaction is completed, water is added to form a mixed solution. The mixed solution is then sonicated to produce a solid precipitation. The solid is filtered to obtain a white solid. The solid is dried to obtain the aminohydrolysate.

[0043] Further:

[0044] In step a, the aminolysis reaction is carried out at a temperature of 100°C for 3 hours. The aminolysis reaction is tracked by silica gel thin-layer chromatography. The aminolysis reaction ends when the silica gel thin-layer chromatographic spots of substituted or unsubstituted phthalic anhydrides in the reaction system disappear.

[0045] Furthermore, step b includes:

[0046] The aminohydrolysate was dissolved in thionyl chloride and then heated under reflux to carry out an acylation reaction to obtain a yellow oily intermediate A, wherein the acylation reaction lasted for 2 hours.

[0047] Furthermore, step c includes:

[0048] In putrescine, tetrahydrofuran was added as a solvent, and the acylamine chloride hydrolysate, intermediate A, was added dropwise under ice bath conditions for an ice bath reaction, followed by a room temperature reaction. After the reaction was completed, a room temperature reaction product was obtained. The room temperature reaction product was then purified by a pulping method to obtain a pale yellow powder solid containing the isoindoline ketone derivative with the structure shown in Formula I.

[0049] Further:

[0050] In step c, the ice bath reaction lasts for 5 minutes, and the room temperature reaction lasts for 2 hours. The room temperature reaction is tracked by silica gel thin-layer chromatography. The room temperature reaction ends when the silica gel thin-layer chromatographic spot of the ammonium chloride hydrolysate, i.e., intermediate A, in the reaction system disappears.

[0051] Further:

[0052] The purification method described above involves adding ethyl acetate to the reaction product at room temperature to obtain an ethyl acetate mixture, then stirring the ethyl acetate mixture in a high-speed magnetic stirrer, filtering to remove solid impurities, and then concentrating and drying the clarified liquid obtained from the filtration to obtain the purified isoindolone derivative having the structure shown in Formula I.

[0053] Furthermore, step d includes:

[0054] N,N-dimethylformamide was added as a solvent to a 4-dimethylaminopyridine solution of cystamine, and the acylamine chloride hydrolysate, intermediate A, was slowly added dropwise under ice bath conditions. The reaction was first carried out in an ice bath and then at room temperature. After the reaction was completed, a small amount of water was added to quench the reaction. The product was then extracted with ethyl acetate, the organic phase was collected, dried, and concentrated to obtain a deep yellow crude product. The crude product was then purified by silica gel column chromatography to obtain a yellow powdery solid, which is the isoindololinone derivative with the structure shown in Formula II.

[0055] Further:

[0056] In step d, the ice bath reaction lasts for 5 minutes, the room temperature reaction lasts for 3 hours, and both the ice bath reaction and the room temperature reaction are carried out under nitrogen protection.

[0057] The crude product is purified by silica gel column chromatography using a mixed solvent of dichloromethane and methanol in a ratio of 25:1.

[0058] Furthermore, step e, i.e., the first group derivatization reaction, includes:

[0059] Equimolar amounts of the isoindolone derivative having the structure shown in Formula II were mixed with triethylamine and acetonitrile and cooled to -20°C. The reactor was then evacuated, and an equimolar ratio of trifluoromethyliodoline was added. Sodium thiosulfate was then added dropwise to the reactor. The reaction was first carried out in an ice bath and then at room temperature. When the reaction reached its endpoint, an appropriate amount of water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was then collected, dried, and concentrated to obtain the crude product. The pure crude product was separated by column chromatography to obtain the isoindolone derivative having the structure shown in Formula III as a brown solid powder.

[0060] Further:

[0061] In step e, the ice bath reaction lasts for 30 minutes, and the room temperature reaction is tracked by silica gel thin-layer chromatography. The room temperature reaction ends when the silica gel thin-layer chromatographic spot of the isoindoline ketone derivative with the structure shown in Formula II disappears.

[0062] The column chromatography method separates the solid phase as 300-mesh silica gel, and the eluent is a mixture of ethyl acetate and petroleum ether in a ratio of 2:1.

[0063] Furthermore, step f includes:

[0064] Dichloromethane was added to putrescine as a solvent, and 1.5 equivalents of tert-butylcarbonyl (abbreviation: (BOC)2O) were added dropwise under ice bath conditions. The reaction was carried out at 0°C for 12 hours. After the reaction was completed, the mixture was extracted with water and ethyl acetate, the organic phase was collected, dried and concentrated to obtain a yellow oily liquid, namely tert-butylcarbonyl-protected putrescine.

[0065] Furthermore, step g includes:

[0066] Dichloromethane was added to the hydrolysate and an equimolar amount of tert-butylcarbonyl-protected putrescine to completely dissolve it. A catalyst composed of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and 1-hydroxybenzotriazole was added dropwise at 0°C to carry out a catalytic reaction in an ice bath. Then, the mixture was raised to room temperature and stirred for a room temperature stirring reaction. After the reaction was completed, the mixture was washed with a saturated sodium bicarbonate aqueous solution, allowed to stand and separate into layers, and the organic phase was taken. After removing the dichloromethane from the organic phase, a crude product was obtained. The crude product was subjected to column chromatography to obtain a light yellow solid, namely intermediate B.

[0067] Further:

[0068] In step g, the catalytic reaction under ice bath lasts for 30 minutes, and the stirring reaction at room temperature lasts for 18 hours. The stirring reaction at room temperature is tracked by silica gel thin-layer chromatography. The stirring reaction at room temperature ends when the silica gel thin-layer chromatographic spots of the aminohydrolysate in the reaction system disappear.

[0069] The column chromatography uses 300-mesh silica gel as the packing material and a mixture of dichloromethane and methanol in a ratio of 50:1 as the eluent.

[0070] Furthermore, step h includes:

[0071] A dichloromethane solution containing 20% ​​(v / v) trifluoroacetic acid was added to intermediate B, and the second group derivatization reaction was carried out under stirring. After the reaction was completed, the reaction system was adjusted to weak alkalinity with a 15% (w / v) sodium hydroxide aqueous solution. Then, the system was extracted with a mixture of ethyl acetate and water. The organic phase was collected, filtered, dried, and concentrated to obtain a yellow solid product, namely the isoindolone derivative having the structure shown in Formula IV.

[0072] Further:

[0073] The second group derivatization reaction takes 3 to 4 hours and is tracked by silica gel thin-layer chromatography. The second group derivatization reaction ends when the silica gel thin-layer chromatographic spot of intermediate B in the reaction system disappears.

[0074] Furthermore, the present invention also provides an application of isoindolinone derivatives.

[0075] The application involves using compounds having the structures shown in Formula I, or Formula II, or Formula III, or Formula IV for the prevention and control of tomato gray mold, rapeseed sclerotinia stem rot, wheat scab, and wheat sheath blight.

[0076] Compared with the prior art, the beneficial effects and significant progress of the present invention are as follows:

[0077] First, this invention provides a novel series of isoindolinone derivatives with amide bonds as the core structure. These novel isoindolinone derivatives are synthesized from substituted or unsubstituted isoindolinone groups, amino acids, biogenic amines, etc., as raw materials. They not only have excellent control effects on a variety of plant diseases, but also exhibit good biocompatibility because amino acids and biogenic amines are both basic substances of life. In particular, isoindolinone as the parent core structure can enhance its lipid solubility, amino acid derivatives as linking arms can improve its stability, and biogenic amines such as cystamine, putrescine, histamine, etc. as introducing groups can promote the autophagy physiological phenomenon of biological cells. Its innovative mechanism of action opens up new ideas and methods for the application of isoindolinone structural unit compounds in the agricultural field.

[0078] Secondly, since the isoindololinone derivatives provided by this invention are prepared by reacting substituted or unsubstituted phthalic anhydride with amino acids to generate compounds with isoindololinone structural units, and then organically combining them with biogenic amines, the preparation method is relatively simple, the raw materials are readily available and the cost is low, it is easy to industrialize, the yield is high, the economy is good, and it has great value for promotion and application. Detailed Implementation

[0079] To make the objectives, technical solutions, beneficial effects and significant progress of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the reaction formulas provided in the embodiments of the present invention. Obviously, all the described embodiments are only some embodiments of the present invention, and not all embodiments.

[0080] Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without inventive effort are within the scope of protection of this invention.

[0081] It should be noted that:

[0082] The terms "firstly," "secondly," etc., used in the specification, claims, and embodiments of this invention are merely for distinguishing different objects and not for describing a specific order. Furthermore, the term "comprising" and any variations thereof are intended to cover non-exclusive inclusion. For example, a process, method, system, product, or device that includes a series of steps or units is not limited to the listed steps or units, but may optionally include steps or units not listed, or may optionally include other steps or units inherent to these processes, methods, products, or devices.

[0083] It should be understood that some basic operational terms commonly used in the art, such as "heating," "stirring," "mixing," "dissolving," "washing," "filtering," and "drying," are used in the description of the embodiments of the present invention. These terms should be interpreted broadly, and can refer to routine operations performed using various conventional equipment and instruments in the art, as well as operations performed using the latest equipment, such as programmed operations and unmanned automatic operations. Unless otherwise explicitly limited, those skilled in the art can understand the specific meaning of the above terms in the present invention according to the specific circumstances and adopt specific operating methods to achieve their operational objectives.

[0084] It should also be noted that:

[0085] The following specific embodiments can be combined with each other. The same or similar concepts or processes may not be described again in some embodiments. In addition, the raw materials, auxiliary materials and reaction equipment and facilities involved in the following specific embodiments are all commercially available.

[0086] The technical solution of the present invention will now be described in detail with reference to specific embodiments.

[0087] Example 1

[0088] This embodiment provides an isoindolinone derivative.

[0089] An isoindolinone derivative, which is a compound having a structure as shown in Formula I, or Formula II, or Formula III, or Formula IV:

[0090]

[0091] in:

[0092] R 1 It is any one of hydrogen, fluorine, chlorine, nitro, methoxy, methyl, or amino, or R 1 For substituted and unsubstituted phenyl groups, or R 1 It can be a halogenated or non-halogenated five- or six-membered heterocyclic group containing oxygen, nitrogen, and sulfur;

[0093] R 2 For C1~C 10 saturated alkyl groups, or R 2 For substituted or unsubstituted phenyl groups, or R 2 It is a five- or six-membered heterocyclic group containing oxygen, sulfur, and nitrogen, or R 2 for Group and R therein a It is any one of halophenyl, benzyl, pyridyl, furanyl, pyrimidinyl or five-membered cycloyl group;

[0094] R 3 It is a thiol group, or R 3 -(CH2) n NH2- group and n is 1 to 5, or R 3 for Group and R therein b It is sulfonyl or trifluoromethyl;

[0095] R c It is methyl or halophenyl.

[0096] Preferred, R 1 In substituted and unsubstituted phenyl groups, the substituted phenyl group is a halophenyl or Alternatively, it could be a trifluoromethyl-substituted phenyl group.

[0097] Preferred, R 1 In this context, the halophenyl group is...

[0098] Preferred, R 1 It is a halogenated or non-halogenated five- or six-membered heterocyclic group containing oxygen, nitrogen, and sulfur, which is a halogenated or non-halogenated pyridinyl, pyrimidinyl, or furanyl group.

[0099] Preferred:

[0100] R 1 In pyridinyl, pyrimidinyl, or furanyl groups that are halogenated or non-halogenated, the halogenated pyridinyl group is: Its halopyrimidine group is

[0101] Preferred:

[0102] R 2 For C1~C 10 The saturated alkyl group is -CH2- or -(CH2)2- or -(CH2). 10 -;

[0103] R 2 In substituted or unsubstituted phenyl groups, the substituted phenyl group is... Or for Or perhaps

[0104] R 2 Among five- and six-membered heterocyclic groups containing oxygen, sulfur, and nitrogen, the nitrogen-containing five-membered heterocyclic group is: Its nitrogen-containing six-membered heterocyclic group is

[0105] Preferred:

[0106] R 3 -(CH2) n The NH2- group is either -CH2NH2- or -(CH2)2NH2-.

[0107] From the above description, it can be seen that:

[0108] The isoindolone derivatives provided in this embodiment are a novel series of isoindolone derivatives prepared by organically combining isoindolone structural unit compounds with amino acids and biogenic amines. Experiments have shown that this novel series of isoindolone derivatives exhibits good biocompatibility and excellent properties in promoting autophagy in biological cells. They can be used for the prevention and control of various plant diseases, thus opening up new ways and methods for the application of isoindolone structural unit compounds in the agricultural field.

[0109] Example 2

[0110] This embodiment provides a method for preparing the isoindoline ketone derivative described in Example 1.

[0111] The method for preparing isoindolinone derivatives provided in this embodiment includes steps a~b~c, or steps a~b~d, or steps a~b~d~e, or steps a~g~h in the following synthetic route, wherein step f is included before step g;

[0112] Synthesis route:

[0113]

[0114] in:

[0115] Step a involves reacting substituted or unsubstituted phthalic anhydride with an amino acid to obtain an aminolysis product;

[0116] Step b involves the acylation reaction of the aminohydrolysate with thionyl chloride under reflux to form the acylated aminohydrolysate, i.e., intermediate A;

[0117] Step c involves reacting the acylamine chloride hydrolysate, i.e., intermediate A, with putrescine to obtain an isoindololinone derivative having the structure shown in Formula I.

[0118] Step d involves reacting the ammonium chloride hydrolysate, intermediate A, with cystamine to obtain an isoindololinone derivative with the structure shown in Formula II.

[0119] Step e involves the isoindoline ketone derivative having the structure shown in Formula II undergoing a first group derivatization reaction to obtain an isoindoline ketone derivative having the structure shown in Formula III.

[0120] Step g involves the reaction of the amino hydrolysate with the tert-butylcarbonyl-protected putrescine obtained in step f (i.e., the amino protection reaction) under the action of a catalyst to obtain intermediate B.

[0121] Step h involves the intermediate B undergoing a second group derivatization reaction to obtain an isoindolinone derivative with the structure shown in Formula IV.

[0122] Furthermore, step a above includes:

[0123] Amine hydrolysis was carried out in glacial acetic acid with substituted or unsubstituted phthalic anhydride and an equimolar amount of amino acid. After the reaction was completed, water was added to form a mixed solution. The mixed solution was then sonicated to produce a solid precipitation. The solid was filtered to obtain a white solid. The solid was dried to obtain the aminohydrolysate.

[0124] Further:

[0125] In step a above, the aminolysis reaction is carried out at a temperature of 100°C for 3 hours. The aminolysis reaction is tracked by silica gel thin-layer chromatography. The aminolysis reaction ends when the silica gel thin-layer chromatographic spots of substituted or unsubstituted phthalic anhydrides in the reaction system disappear.

[0126] Furthermore, step b above includes:

[0127] The aminohydrolysate was dissolved in thionyl chloride and then heated under reflux to carry out an acylation reaction to obtain a yellow oily intermediate A. The acylation reaction lasted for 2 hours.

[0128] Furthermore, step c above includes:

[0129] In putrescine, tetrahydrofuran was added as a solvent, and the acylamine chloride hydrolysate, intermediate A, was added dropwise under ice bath conditions to carry out the ice bath reaction, followed by a room temperature reaction. After the reaction was completed, the room temperature reaction product was obtained. Then, the room temperature reaction product was purified by a pulping method to obtain a pale yellow powder solid isoindolone derivative with the structure shown in Formula I.

[0130] Further:

[0131] In step c above, the ice bath reaction lasts for 5 minutes, and the room temperature reaction lasts for 2 hours. The room temperature reaction is tracked by silica gel thin-layer chromatography. The room temperature reaction ends when the silica gel thin-layer chromatographic spot of the ammonium chloride hydrolysate, i.e., intermediate A, disappears.

[0132] Further:

[0133] Purification by pulping involves adding ethyl acetate to the reaction product at room temperature to obtain an ethyl acetate mixture, then stirring the ethyl acetate mixture in a high-speed magnetic stirrer, filtering to remove solid impurities, and then concentrating and drying the clarified liquid obtained by filtration to obtain the purified isoindolone derivative with the structure shown in Formula I.

[0134] Furthermore, step d above includes:

[0135] N,N-dimethylformamide was added as a solvent to a cystamine solution containing 4-dimethylaminopyridine, and the intermediate A, an acylamine chloride hydrolysate, was slowly added dropwise under ice bath conditions. The reaction was first carried out in an ice bath and then at room temperature. After the reaction was completed, a small amount of water was added to quench the reaction. The product was then extracted with ethyl acetate, the organic phase was collected, dried, and concentrated to obtain a deep yellow crude product. The crude product was then purified by silica gel column chromatography to obtain a yellow powdery solid, which is an isoindololinone derivative with the structure shown in Formula II.

[0136] Further:

[0137] In step d above, the ice bath reaction lasts for 5 minutes and the room temperature reaction lasts for 3 hours, and both the ice bath reaction and the room temperature reaction are carried out under nitrogen protection.

[0138] The crude product was purified by silica gel column chromatography using a mixed solvent of dichloromethane and methanol in a ratio of 25:1.

[0139] Furthermore, step e, the first group derivatization reaction, includes:

[0140] An equimolar amount of the isoindolone derivative with the structure shown in Formula II was mixed with triethylamine and acetonitrile and cooled to -20°C. The reactor was then evacuated, and an equimolar amount of trifluoromethyliodoline was added. Sodium thiosulfate was then added dropwise to the reactor. The reaction was first carried out in an ice bath and then at room temperature. When the reaction reached its endpoint, an appropriate amount of water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was then collected, dried, and concentrated to obtain the crude product. The pure crude product was separated by column chromatography to obtain the brown solid powder of the isoindolone derivative with the structure shown in Formula III.

[0141] Further:

[0142] In step e above, the ice bath reaction lasts for 30 minutes, and the room temperature reaction is tracked by silica gel thin-layer chromatography. The room temperature reaction ends when the silica gel thin-layer chromatographic spots of the isoindolone derivative with the structure shown in Formula II disappear.

[0143] The solid phase was separated by column chromatography, which was 300-mesh silica gel. The eluent was a mixture of ethyl acetate and petroleum ether in a ratio of 2:1.

[0144] Furthermore, step f includes:

[0145] Dichloromethane was added to putrescine as a solvent, and 1.5 putrescine equivalents of tert-butylcarbonyl were added dropwise under ice bath conditions. The reaction was carried out at 0°C for 12 hours. After the reaction was completed, the mixture was extracted with water and ethyl acetate, the organic phase was collected, dried and concentrated to obtain a yellow oily liquid, namely putrescine protected by tert-butylcarbonyl.

[0146] Furthermore, step g includes:

[0147] Dichloromethane was added to the aminohydrolysate and an equimolar amount of tert-butylcarbonyl-protected putrescine to completely dissolve the amine. A catalyst composed of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and 1-hydroxybenzotriazole was added dropwise at 0°C to carry out the catalytic reaction under ice bath conditions. The reaction was then carried out by stirring at room temperature. After the reaction was completed, the mixture was washed with a saturated sodium bicarbonate aqueous solution, allowed to stand and separate into layers, and the organic phase was collected. Dichloromethane was removed from the organic phase to obtain the crude product. The crude product was subjected to column chromatography to obtain a light yellow solid, which is intermediate B.

[0148] Furthermore, in step g above, the catalytic reaction under ice bath lasts for 30 minutes, and the reaction under room temperature stirring lasts for 18 hours. The room temperature stirring reaction is tracked by silica gel thin-layer chromatography. When the silica gel thin-layer chromatographic spots of the aminohydrolysate in the reaction system disappear, the room temperature stirring reaction ends. The column chromatography uses 300-mesh silica gel as the packing material and a mixture of dichloromethane and methanol in a ratio of 50:1 as the eluent.

[0149] Furthermore, step h includes:

[0150] A dichloromethane solution containing 20% ​​(v / v) trifluoroacetic acid was added to intermediate B, and the second group derivatization reaction was carried out under stirring. After the reaction was completed, the reaction system was adjusted to weak alkalinity with a 15% (w / v) sodium hydroxide aqueous solution. The mixture of ethyl acetate and water was then used for extraction. The organic phase was collected, filtered, dried, and concentrated to obtain a yellow solid product, which is an isoindololinone derivative with the structure shown in Formula IV.

[0151] Furthermore, the second group derivatization reaction takes 3 to 4 hours, and the second group derivatization reaction is tracked by silica gel thin-layer chromatography. When the silica gel thin-layer chromatographic spot of intermediate B in the reaction system disappears, the second group derivatization reaction is considered to be over.

[0152] From the above description, it can be seen that:

[0153] The isoindolone derivatives provided in this embodiment are prepared by reacting substituted or unsubstituted phthalic anhydride with amino acids to generate compounds with isoindolone structural units, and then organically combining them with biogenic amines. Therefore, the preparation method is relatively simple, the raw materials are readily available and the cost is low, it is easy to industrialize, the yield is high, the economy is good, and it has great value for promotion and application.

[0154] To further aid in understanding the technical solution provided in this embodiment, as well as the specific operation process and the effects that can be obtained, the preparation method will be further explained below through specific examples.

[0155] Case 1: Synthesis of N,N'-(butane-1,4-diyl)bis(2-(1,3-dioxoisoindol-2-yl)-3-phenylpropionamide)

[0156] Synthesis route:

[0157]

[0158] Preparation process:

[0159] Weigh 3 mmol of phthalic anhydride and glycine into a 100 ml round-bottom flask, add 50 ml of glacial acetic acid (AcOH) to dissolve them, set the temperature to 100 °C and heat for 3 hours. Monitor the reaction using silica gel thin-layer chromatography (TLC). When the starting material spot disappears, add 3 times the solvent volume of deionized water to obtain a mixed solution. Sonicate the mixed solution until a solid product appears, filter to obtain a white solid, and dry the solid with an infrared drying instrument to obtain the aminolysis product, i.e., intermediate A, which is 2-(1,3-dioxoisoindol-2-yl)-3-phenylpropionic acid as shown in the above synthetic route. Its molar yield relative to phthalic anhydride is 70%.

[0160] Weigh 3 mmol (0.885 g) of the above intermediate A into a single-necked flask, add an appropriate amount of thionyl chloride (molecular formula: SOCl2), reflux at 80 °C for 3-4 hours, monitor the reaction with silica gel thin-layer chromatography plate, after the reaction is completed, a brown oily liquid is obtained, evaporate the solvent and set aside for later use.

[0161] Another 1.5 mmol (0.132 g) of putrescine was weighed into a three-necked flask, and 50 mL of tetrahydrofuran was added as solvent. 2-(1,3-dioxoisoindol-2-yl)-3-phenylpropionyl chloride was slowly added dropwise under ice bath conditions. After reacting for 5 minutes, the ice bath was removed, and the reaction was carried out at room temperature for 2 hours. The reaction was monitored by TLC. After the reaction was completed, the mixture was purified by slurry to obtain a pale yellow powder solid, which is an isoindolone derivative with the structure shown in Formula I. The molar yield of this derivative relative to intermediate A, 2-(1,3-dioxoisoindol-2-yl)-3-phenylpropionic acid, was 38%.

[0162] Testing and verification:

[0163] 1 H NMR(600MHz,DMSO-d6)δ8.30(s,1H),7.83~7.75(m,9H),7.11(q,J=6.9Hz,8H),4.94(dd,J=12.0,4.6Hz,2H ),3.54(dd,J=14.0,4.5Hz,3H),3.31(dd,J=14.0,12.0Hz,2H),3.08(dp,J=18.4,6.3Hz,6H),1.39(s,5H);

[0164] 13C NMR (101MHz, DMSO-d6) δ167.96,138.25,134.98,129.66,129.20,128.75,128.59,126.95,123.56,54.90,39.27,34.44,26.95;

[0165] HRMS(ESI): m / z caled for C 38 H 34 N4O6[M+H] + 642.2478, found 642.2489.

[0166] Case 2: Synthesis of [2-(1,3-dioxoisoindol-2-yl)-N-(2-mercaptoethyl)acetamide]

[0167] Synthesis route:

[0168]

[0169] Preparation process:

[0170] Weigh 3 mmol of phthalic anhydride and glycine into a 100 ml round-bottom flask, add 50 ml of glacial acetic acid (AcOH) to dissolve them, set the temperature to 100 °C and heat for 3 hours. Monitor the reaction using silica gel thin-layer chromatography (TLC). When the starting material spot, i.e., the phthalic anhydride spot, disappears, add 3 times the solvent volume of deionized water and sonicate the mixture until a solid product appears. Filter to obtain a white solid, dry the solid with an infrared drying instrument to obtain the aminohydrolysate. The molar yield relative to phthalic anhydride is 70%.

[0171] Take 0.707 g (3.41 mmol) of the aminolysis product and add it to a 50 ml single-necked flask. Add an appropriate amount of thionyl chloride (SOCl2, the same below) to dissolve it. Heat to 80 °C and react for 2 hours to obtain a yellow oily substance, i.e., intermediate A.

[0172] Take a 100mL three-necked flask, add 6.82mmol (1.054g) of 2,2'-dithiodimethylbis(ethane-1-amine) (i.e., cystamine) and 2.4mmol (1.6g) of 4-dimethylaminopyridine (DMAP, hereinafter the same), and dissolve it in 10mL of N,N-dimethylformamide (DMF, hereinafter the same). Intermediate A was slowly added dropwise under ice bath conditions. After reacting for 5 minutes, the ice bath was removed and the reaction was carried out at a constant temperature for 3 hours. The reaction required nitrogen protection to prevent interference from oxygen or carbon dioxide in the air, and the reaction system had to be carried out under anhydrous conditions. After the reaction was completed, a small amount of water was added to quench the reaction. The product was extracted with ethyl acetate, the organic phase was collected, dried, and concentrated to obtain a deep yellow crude product. The crude product was purified by silica gel column chromatography with dichloromethane (DCM) and methanol (MeOH) as eluents at a ratio of 25:1 to obtain a yellow powdery solid product, namely an isoindololinone derivative with the structure shown in Formula II. The molar yield was 50% compared to intermediate A.

[0173] Testing and verification:

[0174] 1 H NMR (400MHz, DMSO-d6) δ8.44(s,1H),7.89(ddt,J=14.5,5.7,3.3Hz,8H),4.19(s,2H),3.42~3.35(m,4H),2.77(t,J=6.7Hz,3H),1.23(s,1H);

[0175] 13 C NMR (101MHz, DMSO) δ168.02,166.75,135.09,132.27,123.73,40.62,38.65,37.39;

[0176] HRMS(ESI):m / z caled for C12H 12 N₂O₃S[M+H] + 264.0569, found 264.0571.

[0177] In Cases 3-14 below, the starting materials (not indicated in some synthetic routes) are all intermediates A, which are acyl chloride amino acid derivatives with different groups obtained by aminolysis of phthalic anhydride and different amino acids. Since the synthetic operation steps of intermediate A are similar to those of the corresponding intermediates A in Cases 1 and 2 above, except that the amino acids involved in the reaction are different, the synthetic process of the starting material intermediate A will not be described again in the following cases. Only the synthetic steps of reacting different intermediates A with cystamine to obtain the corresponding isoindolinone derivatives with the structure shown in Formula II will be described.

[0178] Case 3: Synthesis of 11-(1,3-dioxoisoindoline-2-yl)-N-(2-mercaptoethyl)undecanoamide

[0179] Partial synthetic route:

[0180]

[0181] Partial preparation process:

[0182] By replacing intermediate A in Case 2 with 11-(1,3-dioxoisoindoline-2-yl)undecanoyl chloride, and then following the same reaction process and operation steps as in Case 2, 11-(1,3-dioxoisoindoline-2-yl)-N-(2-mercaptoethyl)undecanoamide, which is a pure light yellow solid, can be obtained with a molar yield of 56% relative to intermediate A.

[0183] Testing and verification:

[0184] 1 H NMR (400MHz, DMSO-d6) δ7.97 (t, J=5.5Hz, 1H), 7.91~7.76 (m, 4H), 3.55 (t, J=7.1Hz, 2H), 3.32~3.2 7(m,2H),2.75(t,J=6.7Hz,2H),2.04(t,J=7.3Hz,2H),1.63~1.41(m,3H),1.22(d,J=17.2Hz,11H);

[0185] 13 C NMR (101MHz, DMSO-d6) δ168.37,167.86,138.11,135.03,131.80,129.23,128.76,126.98,123.61,54.71,39.12,37.2434.37,30.12;

[0186] HRMS(ESI): m / z caled for C 21H 30 N₂O₃S[M+H] + 390.1977, found 390.1978.

[0187] Case 4: Synthesis of N-(2-mercaptoethyl)-11-(4-nitro-1,3-dioxoisoindol-2-yl)undecanoamide

[0188] Partial synthetic route:

[0189]

[0190] Partial preparation process:

[0191] By replacing intermediate A in Case 2 with 11-(4-nitro-1,3-dioxoisoindol-2-yl)undecanoyl chloride, and then following the same reaction process and operation steps as in Case 2, a pure light yellow solid can be obtained with a molar yield of 49% relative to intermediate A.

[0192] Testing and verification:

[0193] 1 H NMR (400MHz, DMSO-d6) δ8.26(d,J=8.0Hz,1H),8.15(d,J=7.2Hz,1H),8.05(d,J=7.8Hz,1H),3.54(d,J=7.1Hz,2 H),3.29(s,1H),2.75(t,J=6.6Hz,2H),2.04(t,J=7.3Hz,2H),1.51(d,J=46.5Hz,4H),1.24(d,J=17.9Hz,13H);

[0194] 13 C NMR (101MHz, DMSO) δ172.83,166.54,163.90,144.80,136.61,134.13,128.67,127.23, 123.57,38.49,38.38,37.91,35.86,29.37,29.27,29.15,29.07,28.17,26.74,25.74;

[0195] HRMS(ESI): m / z caled for C 21 H 29 N3O5S[M+H] + 435.1828, found 435.1831.

[0196] Case 5: Synthesis of 3-(1,3-dioxoisoindol-2-yl)-N-(2-mercaptoethyl)propionamide

[0197] Partial synthetic route:

[0198]

[0199] Partial preparation process:

[0200] By replacing intermediate A in Case 2 with 3-(1,3-dioxoisoindol-2-yl)propionyl chloride, and then following the same reaction process and operation steps as in Case 2, a pure product in dark yellow solid with a molar yield of 65% relative to intermediate A can be obtained.

[0201] Testing and verification:

[0202] 1 H NMR (400MHz, DMSO-d6) δ8.18(s,1H),7.97~7.68(m,10H),3.77(t,J=7.2Hz,4H),3.25(q,J=6.3Hz,4H),2.68(t,J=6.7Hz,4H),2.43(t,J=7.2Hz,4H);

[0203] 13 C NMR (101MHz, DMSO) δ170.22,168.15,134.83,132.22,123.49,38.37,37.48,34.90,34.51;

[0204] HRMS(ESI): m / z caled for C 13 H 14 N₂O₃S[M+H] + 278.0725, found 278.0716.

[0205] Case 6: Synthesis of 2-(1,3-dioxoisoindol-2-yl)-N-(2-mercaptoethyl)-3-phenylpropionamide

[0206] Partial synthetic route:

[0207]

[0208] Partial preparation process:

[0209] By replacing intermediate A in Case 2 with 2-(1,3-dioxoisoindol-2-yl)-3-phenylpropionyl chloride, and then following the same reaction process and operation steps as in Case 2, a pure yellow solid can be obtained with a molar yield of 70% relative to intermediate A.

[0210] Testing and verification:

[0211] 1 H NMR (600MHz, DMSO-d6) δ8.36 (t, J = 5.6Hz, 1H), 7.92~7.73 (m, 7H), 7.21~7.01 (m, 5H), 4. 95(dd,J=11.9,4.6Hz,1H),3.31~3.27(m,2H),2.86~2.70(m,J=6.3Hz,2H),1.24(s,1H);

[0212] 13 C NMR (101MHz, DMSO) δ168.37,167.86,138.12,135.03,131.81,129.23,128.76,126.98,123.61,54.72,39.13,37.27,34.38;

[0213] HRMS(ESI): m / z caled for C 19 H 18 N₂O₃S[M+H] + 354.1038, found 354.1042.

[0214] Case 7: Synthesis of 2-(4-chloro-1,3-dioxoisoindol-2-yl)-N-(2-mercaptoethyl)acetamide

[0215] Partial synthetic route:

[0216]

[0217] Partial preparation process:

[0218] By replacing intermediate A in Case 2 with 2-(4-chloro-1,3-dioxoisoindol-2-yl)acetyl chloride, and then following the same reaction process and operation steps as in Case 2, a pure yellow solid can be obtained with a molar yield of 72% relative to intermediate A.

[0219] Testing and verification:

[0220] 1 H NMR (400MHz, DMSO-d6) δ8.44(t,J=5.5Hz,1H),7.87(tt,J=7.3,4.4Hz,4H),4.18(s,2H),3.37(d,J=6.3Hz,2H),2.77(t,J=6.7Hz,2H);

[0221] 13C NMR (101MHz, DMSO) δ166.61,166.56,165.64,136.59,136.28,134.55,130.11,127.99,122.68,40.79,38.66,37.37,30.93;

[0222] HRMS(ESI): m / z caled for C 12 H 11 N₂O₃S[M+H] + 298.0179, found 298.0185.

[0223] Case 8: Synthesis of N-(2-mercaptoethyl)-2-(4-nitro-1,3-dioxoisoindol-2-yl)acetamide

[0224] Partial synthetic route:

[0225]

[0226] Partial preparation process:

[0227] By replacing intermediate A in Case 2 with 2-(4-nitro-1,3-dioxoisoindol-2-yl)acetyl chloride, and then following the same reaction process and operation steps as in Case 2, a pure brownish-yellow solid can be obtained with a molar yield of 56% relative to intermediate A.

[0228] Testing and verification:

[0229] 1 H NMR(600MHz,DMSO-d6)δ8.45(s,1H),8.33(d,J=8.1Hz,1H),8.22(d,J=7.4Hz,2H) ,8.11(d,J=7.8Hz,1H),3.36(d,J=6.4Hz,5H),2.77(t,J=6.8Hz,4H),1.23(s,1H);

[0230] 13 C NMR (101MHz, DMSO-d6) δ169.02,165.78,163.18,144.98,137.30,133.70,129.37,127.83,123.27;

[0231] HRMS(ESI): m / z caled for C 12 H 11 N3O5S[M+H] + 309.0419, found 309.0421.

[0232] Case 9: Synthesis of 2-(4-chloro-1,3-dioxoisoindol-2-yl)-N-(2-mercaptoethyl)-3-phenylpropionamide

[0233] Partial synthetic route:

[0234]

[0235] Partial preparation process:

[0236] By replacing intermediate A in Case 2 with 2-(4-chloro-1,3-dioxoisoindol-2-yl)-3-phenylpropionyl chloride, and then following the same reaction process and operation steps as in Case 2, a pure product in the form of a deep yellow powder solid can be obtained, with a molar yield of 77% relative to intermediate A.

[0237] Testing and verification:

[0238] 1 H NMR(600MHz, DMSO-d6)δ8.40(t,J=5.6Hz,1H),7.92~7.74(m,3H),7.27~6.99(m,5H),4.95(dd,J=11.7,4.7Hz,1H), 3.52(dd,J=14.1,4.7Hz,1H),3.41(s,0H),3.32(s,0H),3.25(dd,J=14.0,11.8Hz,1H),2.82~2.68(m,J=6.4Hz,2H);

[0239] 13 C NMR(101MHz,DMSO-d6)δ168.14,166.40,165.38,138.05,136.64,136.37,134.03,130.01, 129.24,128.81,127.42,127.03,122.64,56.54,54.81,39.11,37.24,34.29,30.12,19.08;

[0240] HRMS(ESI): m / z caled for C 19 H 17 N₂O₃S[M+H] + 388.0648, found 388.0655.

[0241] Case 10: Synthesis of 4-chloro-3-(1,3-dioxoisoindol-2-yl)-N-(2-mercaptoethyl)benzamide

[0242] Partial synthetic route:

[0243]

[0244] Partial preparation process:

[0245] By replacing intermediate A in Case 2 with 2-(4-chloro-1,3-dioxoisoindol-2-yl)-3-phenylformyl chloride, and then following the same reaction process and operation steps as in Case 2, a pure pink powder solid can be obtained with a molar yield of 43% relative to intermediate A.

[0246] Testing and verification:

[0247] 1 H NMR (600MHz, DMSO-d6) δ9.09(t,J=5.4Hz,1H),8.16(d,J=2.1Hz,1H),8.10(dd,J=8.5,2.1Hz,1H),8.04(dd,J=5 .5,3.0Hz,2H),7.97(td,J=5.2,2.0Hz,2H),7.83(d,J=8.5Hz,1H),3.60(q,J=6.4Hz,2H),2.98(t,J=6.8Hz,2H);

[0248] 13 C NMR (101MHz, DMSO-d6) δ166.76,164.96,135.71,134.80,131.86,131.14,130.43,130.23,130.05,124.39,39.36,37.44;

[0249] HRMS(ESI): m / z caled for C 17 H 23 N₂O₃S[M+H] + 360.0335, found 360.0336.

[0250] Case 11: Synthesis of 2-(5-chloro-1,3-dioxoisoindol-2-yl)-N-(2-mercaptoethyl)acetamide

[0251] Partial synthetic route:

[0252]

[0253] Partial preparation process:

[0254] By replacing intermediate A in Case 2 with 2-(5-chloro-1,3-dioxoisoindol-2-yl)acetyl chloride, and then following the same reaction process and operation steps as in Case 2, a pure yellow solid can be obtained with a molar yield of 78% relative to intermediate A.

[0255] Testing and verification:

[0256] 1 H NMR (600MHz, DMSO-d6) δ8.43(s,1H),8.01(s,1H),7.92(s,3H),4.19(s,3H),3.36(d,J=6.3Hz,3H),2.77(t,J=6.8Hz,4H),1.23(s,1H);

[0257] 13 C NMR (101MHz, DMSO-d6) δ167.09,166.75,166.55,139.94,134.87,134.23,130.78,125.49,123.90,40.79,38.63,37.36.;

[0258] HRMS(ESI): m / z caled for C 12 H 11 N₂O₃S[M+H] + 298.0179, found 298.0182.

[0259] Case 12: Synthesis of 6-(1,3-dioxoisoindol-2-yl)-N-(2-mercaptoethyl)nicotinamide

[0260] Partial synthetic route:

[0261]

[0262] Partial preparation process:

[0263] By replacing intermediate A in Case 2 with 6-(1,3-dioxoisoindoline-2-yl)nicotinamide chloride, and then following the same reaction process and operation steps as in Case 2, a pure light yellow solid can be obtained with a molar yield of 57% relative to intermediate A.

[0264] Testing and verification:

[0265] 1H NMR (400MHz, DMSO-d6) δ9.07~9.04(m,1H),9.01(t,J=5.5Hz,1H),8.42(dd,J=8.3,2.4Hz,1H),8.11(d,J=1.6Hz,1 H), 8.06~7.96 (m, 2H), 7.67 (d, J = 8.4Hz, 1H), 3.65 (q, J = 6.4Hz, 2H), 3.01 (t, J = 6.8Hz, 2H), 1.26 (d, J = 7.0Hz, 1H);

[0266] 13 C NMR (101MHz, DMSO-d6) δ166.71,164.83,148.71,148.33,138.09,135.64,131.88,130.38,124.31,122.80,39.37,37.53

[0267] HRMS(ESI): m / z caled for C 16 H 13 N3O3S[M+H] + 327.0678, found 327.0688.

[0268] Case 13: Synthesis of 6-(5-chloro-1,3-dioxoisoindol-2-yl)-N-(2-mercaptoethyl)nicotinamide

[0269] Partial synthetic route:

[0270]

[0271] Partial preparation process:

[0272] By replacing intermediate A in Case 2 with 6-(5-chloro-1,3-dioxoisoindol-2-yl)nicotinic acid chloride, and then following the same reaction process and operation steps as in Case 2, a pure light yellow solid can be obtained with a molar yield of 68% relative to intermediate A.

[0273] Testing and verification:

[0274] 1 H NMR (400MHz, DMSO-d6) δ9.05(d,J=2.3Hz,1H),9.01(t,J=5.5Hz,1H),8.42(dd,J=8.3,2.4Hz,1H),8.02~ 7.91(m,3H),7.68(d,J=8.3Hz,1H),3.65(q,J=6.4Hz,2H),3.01(t,J=6.8Hz,2H),1.26(d,J=5.8Hz,1H);

[0275] 13 C NMR(101MHz,DMSO-d6)δ165.35,164.79,164.36,148.71,148.01,138.14,137.94,13 6.93,136.73,134.22,130.67,130.47,127.61,123.17,122.81,53.17,39.34,37.48;

[0276] HRMS(ESI): m / z caled for C 16 H 12 N3O3S[M+H] + 361.0288, found 361.0293.

[0277] Case 14: Synthesis of 6-(4-chloro-1,3-dioxoisoindol-2-yl)-N-(2-mercaptoethyl)nicotinamide

[0278] Partial synthetic route:

[0279]

[0280] Partial preparation process:

[0281] By replacing intermediate A in Case 2 with 6-(4-chloro-1,3-dioxoisoindol-2-yl)nicotinic acid chloride, and then following the same reaction process and operation steps as in Case 2, a pure light yellow solid can be obtained with a molar yield of 53% relative to intermediate A.

[0282] Testing and verification:

[0283] 1 H NMR (400MHz, DMSO-d6) δ9.05(d,J=2.3Hz,1H),9.01(t,J=5.6Hz,1H),8.42(dd,J=8.3,2.4Hz,1H),8.02~ 7.91(m,3H),7.68(d,J=8.3Hz,1H),3.65(q,J=6.4Hz,2H),3.01(t,J=6.8Hz,2H),1.26(d,J=5.8Hz,1H);

[0284] HRMS(ESI): m / z caled for C 16 H 12 N3O3S[M+H] + 361.0288, found 361.0299.

[0285] Furthermore, by following the preparation methods described in Examples 2-14, substituted or unsubstituted phthalic anhydrides and different amino acid raw materials can be used to prepare the structures listed in Table 1 with different R values. 1 R 2 R 3 The resulting isoindolinone derivatives 19–33, 100–132, 199–231, and 298–330 have structures as shown in Formula II.

[0286] Table 1

[0287]

[0288]

[0289]

[0290]

[0291]

[0292]

[0293]

[0294]

[0295]

[0296] In Case 15, the synthesis of 2-(1,3-dioxoisoindol-2-yl)-N-(2-(trifluoromethyl)thio)ethyl)acetamide, since the preparation of the starting material, i.e., the isoindolone derivative with the structure shown in Formula II, is the same as the preparation method of the corresponding products in Cases 2 to 13, it will not be repeated. In this case, only the first group derivatization reaction in the process of preparing the isoindolone derivative with the structure shown in Formula III will be described.

[0297] First group derivatization reaction route:

[0298]

[0299] Preparation process:

[0300] Equal molar amounts of the isoindololinone derivative with the structure shown in Formula II were weighed and reacted with a solution of triethylamine (Et3N) and acetonitrile (MeCN). The mixture was cooled to -20°C, and the reactor was evacuated. An equimolar ratio of trifluoromethyliodoline (CF3I) was added, and sodium thiosulfate (Na2S2O4) was added dropwise to the reactor. After reacting for 0.5 hours, the mixture was allowed to react at room temperature, and the reaction process was monitored by TLC. After the starting material spot disappeared, an appropriate amount of water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was collected, dried, and concentrated to obtain the crude product. The crude product was purified by column chromatography (solid phase: 300-mesh silica gel). The eluent was ethyl acetate (EA): petroleum ether (PE) = 2:1, yielding a brown solid powder, which is the isoindololinone derivative with the structure shown in Formula III. The molar yield relative to the reactants was 76%.

[0301] Testing and verification:

[0302] 1 H NMR (400MHz, DMSO-d6) δ8.44(s,1H),7.89(ddt,J=14.5,5.7,3.3Hz,8H),4.19(s,2H),3.42~3.35(m,4H),2.77(t,J=6.7Hz,3H),1.23(s,1H);

[0303] 13 C NMR (101MHz, DMSO) δ168.02, 166.75, 135.09, 132.27, 123.73, 40.62, 38.65, 37.39, 30.74;

[0304] HRMS(ESI):m / z caled for C12H 12 N₂O₃S[M+H] + 333.2972, found 333.2974.

[0305] Case 16: Synthesis of N-(4-aminobutyl)-2-(1,3-dioxoisoindol-2-yl)acetamide

[0306] Synthesis route:

[0307]

[0308] Preparation process:

[0309] In this case, the preparation of the aminohydrolysate, namely 2-(1,3-dioxoisoindol-2-yl)acetic acid, is the same as in Case 2, therefore, it will not be described again.

[0310] 10 mmol (1.023 ml) of putrescine was pipetted into a 250 ml round-bottom flask. Dichloromethane (DCM) was used as the solvent. 1.5 equivalents (3.273 g) and 15 mmol of tert-butylcarbonyl ((BOC)₂O) were added dropwise under ice bath conditions as a protecting group. The reaction was carried out at 0 °C for 12 hours. After the reaction was completed, the mixture was extracted with water and ethyl acetate. The organic phase was collected, dried, and concentrated to obtain a yellow oily liquid, namely tert-butylcarbonyl-protected putrescine. The molar yield was 90% relative to putrescine.

[0311] Weigh 3.98 mmol (0.8 g) of 2-(1,3-dioxoisoindol-2-yl)acetic acid and an equivalent amount (0.815 g) of tert-butylcarbonyl-protected putrescine. Dissolve them completely in 20 mL of dichloromethane (DCM). At 0 °C, slowly add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) / 1-hydroxybenzotriazole (HOBt) as a catalyst. After reacting for 30 minutes, transfer... The mixture was stirred at room temperature for 18 hours. The reaction was monitored by TLC. After the reaction was stopped, the mixture was washed three times with a saturated sodium bicarbonate (NaHCO3) aqueous solution, allowed to stand for separation, and the DCM phase was evaporated to dryness to obtain the crude product. The crude product was eluted by column chromatography with a mixture of dichloromethane (DCM):methanol (MeOH) = 50:1 to obtain a light yellow solid, which is intermediate B. The molar yield of intermediate B relative to 2-(1,3-dioxoisoindol-2-yl)acetic acid was 85%.

[0312] The pure intermediate B was added to a DCM solution containing 20% ​​trifluoroacetic acid (TFA) and stirred for 3-4 hours. The reaction was monitored by TLC. After the starting material spot disappeared, the reaction system was adjusted to weakly alkaline with 15% sodium hydroxide (NaOH) aqueous solution. Then, it was extracted with ethyl acetate (EA) and water. The organic phase was dried, filtered, and concentrated to obtain a yellow solid product, which is an isoindololinone derivative with the structure shown in Formula IV. The molar yield of this product relative to intermediate B was 63%.

[0313] Testing and verification:

[0314] 1 H NMR (600MHz, DMSO-d6) δ8.24 (s, 1H), 7.97 (s, 2H), 7.89 (ddt, J = 21.3, 5.6, 3.1Hz, 4H), 4.1 7(s,2H),3.06(d,J=5.6Hz,2H),2.79(t,J=6.7Hz,2H),1.60(t,J=6.9Hz,2H),1.39(s,2H);

[0315] HRMS(ESI): m / z caled for C 14 H 17 N3O3[M+H] + 275.1270, found 275.1265.

[0316] In Cases 17-19 below, the starting materials (not indicated in some synthetic routes) are different intermediates B. Since the synthetic operation steps of different intermediates B are similar to those of intermediate B in Case 15 above, the only difference is that different amino acids are involved in the reaction, resulting in different aminohydrolysates. Therefore, in the following cases, the synthetic process of the aminohydrolysates will not be described again, but only the synthetic steps of reacting different intermediates B with cystamine to obtain the corresponding isoindolinone derivatives with the structure shown in Formula II will be explained.

[0317] Case 17: Synthesis of N-(4-aminobutyl)-2-(4-chloro-1,3-dioxoisoindol-2-yl)acetamide

[0318] Partial synthetic route:

[0319]

[0320] Partial preparation process:

[0321] Intermediate B from Example 14 was replaced with tert-butyl(4-(2-(4-chloro-1,3-dioxoisoindol-2-yl)acetamyl)carbamate, and other reaction conditions were the same as in Example 14. The final product was a pale yellow solid powder with a yield of 80%.

[0322] Testing and verification:

[0323] 1 H NMR (400MHz, DMSO-d6) δ8.27 (t, J = 5.8Hz, 1H), 7.99~7.82 (m, 3H), 7.67 (s, 3H), 4. 16(s,2H),3.08(q,J=6.3Hz,2H),2.79(s,2H),1.48(dq,J=20.5,7.7,6.9Hz,4H);

[0324] 13 C NMR (101MHz, DMSO-d6) δ166.70,166.32,165.73,136.64,136.31,134.61,130.11,128.04,122.69,40.85,39.06,38.66,26.56,25.01;

[0325] HRMS(ESI): m / z caled for C 14 H 16 N3O3[M+H] + 309.0880, found 309.0891.

[0326] Case 18: Synthesis of N-(4-aminobutyl)-2-(5-nitro-1,3-dioxoisoindole-2-yl)acetamide

[0327] Partial synthetic route:

[0328]

[0329] Partial preparation process:

[0330] Intermediate B from Example 14 was replaced with tert-butyl(4-(2-(4-nitro-1,3-dioxoisoindole-2-yl)acetamido)carbamate, and other reaction conditions were the same as in Example 14. The final product was a pale yellow solid powder with a yield of 69%.

[0331] Testing and verification:

[0332] 1 H NMR (400MHz, DMSO-d6) δ8.37(d,J=8.0Hz,1H),8.33(t,J=5.7Hz,1H),8.25(d,J=7.5Hz,1H),8.14(t,J=7.8Hz ,1H),7.79~7.66(m,3H),4.22(s,2H),3.12(q,J=6.4Hz,2H),2.82(s,2H),1.51(dp,J=21.1,7.8,7.2Hz,5H);

[0333] 13 C NMR (101MHz, DMSO-d6) δ166.07,163.49,144.86,137.03,134.05,129.00,127.51,123.51,41.11,39.02,38.64,26.52,24.97;

[0334] HRMS(ESI): m / z caled for C 14 H 16 N4O5[M+H] + 320.1121, found 320.1117.

[0335] Case 19: Synthesis of N-(4-aminobutyl)-2-(5-nitro-1,3-dioxoisoindole-2-yl)acetamide

[0336] Partial synthetic route:

[0337]

[0338] Partial preparation process:

[0339] Intermediate B from Example 14 was replaced with tert-butyl(4-(2-(5-nitro-1,3-dioxoisoindole-2-yl)acetamyl)carbamate, and other reaction conditions were the same as in Example 14. The final product was a pale yellow solid powder with a yield of 78%.

[0340] Testing and verification:

[0341] 1 H NMR (400MHz, DMSO-d6) δ8.69(dd,J=8.2,2.1Hz,1H),8.56(d,J=2.1Hz,1H),8.36(t,J=5.8Hz,1H),8.20(d,J= 8.1Hz,1H),7.76(s,3H),4.25(s,2H),3.11(q,J=6.4Hz,2H),2.81(s,2H),1.51(dp,J=21.8,7.8,7.3Hz,5H);

[0342] 13 C NMR (101MHz, DMSO-d6) δ166.43,166.17,166.09,152.09,136.81,133.57,130.36,125.25,118.57,41.13,39.03,38.66,26.52,24.98;

[0343] HRMS(ESI): m / z caled for C 14 H 16 N4O5[M+H] + 320.1121, found 320.1143.

[0344] Furthermore, by imitating the preparation methods described in Examples 16-19, different intermediates B can be used to prepare the R structures listed in Table 2. 1 R 2 R 3 The isoindololinone derivatives 34–99, 133–198, 232–297, and 331–396 with structures as shown in Formula IV, wherein α in the table represents Group, β represents Group.

[0345] Table 2

[0346]

[0347]

[0348]

[0349]

[0350]

[0351]

[0352]

[0353]

[0354]

[0355]

[0356] Example 3

[0357] This embodiment provides an application of the isoindolinone derivative described in Example 1.

[0358] The application described in this embodiment is to use the isoindoline ketone derivatives with the structures shown in Formula I, or Formula II, or Formula III, or Formula IV as described in Example 1 for the prevention and control of tomato gray mold, rapeseed sclerotinia rot, wheat scab, and wheat sheath blight.

[0359] To further aid in understanding the technical solution provided in this embodiment, as well as the specific operation process and technical effects that can be obtained, the application will be further explained below through specific effect examples. Specifically, experiments will be conducted using isoindolinone derivatives with different product numbers listed in Tables 1 and 2 obtained by the preparation method described in Example 2.

[0360] Example 1: The control effect of isoindoline ketone derivatives with structures shown in Formula I, Formula II, Formula III, or Formula IV on tomato leaves, taking tomato gray mold as an example.

[0361] Test method:

[0362] Weigh 10 mg of each of the isoindolinone derivatives listed in Tables 1 and 2 with different product numbers and dissolve them in 1 ml of dimethyl sulfoxide to prepare a stock solution of 10,000 μg / ml. Then dilute the stock solution with an aqueous solution containing 0.1% Triton X-100 to prepare a solution of 200 μg / ml.

[0363] In the treatment trial, fresh tomato leaves were inoculated with 5mm gray mold cakes that had been cultured for 3 days. After the leaves were fully infected, they were immersed in the drug solution and then placed in a light incubator for cultivation. Each treatment was repeated 3 times. The control group was an aqueous solution containing 0.1% TritiumX-100. The area of ​​the fungal patch was measured 5 days after the application of the drug, and the treatment efficacy (%) was calculated.

[0364] In the protection test, fresh tomato leaves were immersed in the above-mentioned solution for 5 seconds, then removed and allowed to air dry naturally. After inoculating with 5mm gray mold cakes that had been cultured for 3 days, the leaves were placed in a constant temperature and light incubator at 25℃ for 8 hours of light exposure. The protective efficacy (%) was then observed.

[0365] Experimental results:

[0366] The experimental results are listed in Table 3, where the values ​​before and after “ / ” correspond to the therapeutic or protective efficacy of products with two different substituents in the product with that number in Table 2 or Table 3.

[0367] Table 3

[0368]

[0369]

[0370]

[0371]

[0372] The data listed in Table 3 shows that:

[0373] The isoindoline ketone derivatives with different product numbers listed in Tables 1 and 2 all showed certain therapeutic and protective effects against tomato gray mold.

[0374] Example 2: The control effect of isoindoline ketone derivatives with structures shown in Formula I, Formula II, Formula III, or Formula IV on rapeseed leaves, taking Sclerotinia sclerotiorum as an example.

[0375] The experimental method and the concentration of the pesticide used were the same as those in Example 1 above. The rapeseed leaves used were collected from the plants in the field. Five days after the rapeseed leaves were treated with the pesticide, the area of ​​lesions on the leaves was measured and the control effect (%) was calculated.

[0376] The experimental results are listed in Table 4.

[0377] Table 4

[0378]

[0379] As can be seen from the data listed in Table 4:

[0380] Some of the isoindoline ketone derivatives with different product numbers listed in Tables 1 and 2 also showed certain therapeutic and protective effects against sclerotinia stem rot in rapeseed.

[0381] It should be noted that:

[0382] Although Table 4 only lists the test results of some of the products in Tables 1 and 2, based on the actual test results, it can be seen that other isoindoline ketone derivatives with different numbers listed in Tables 1 and 2 also show certain therapeutic and protective effects against sclerotinia stem rot in rapeseed. For the sake of brevity, these are not explained in more detail here.

[0383] Example 3: The control effect of isoindololinone derivatives with structures shown in Formula I, Formula II, Formula III, or Formula IV on wheat seedlings, taking Fusarium graminearum as an example.

[0384] The experimental method and the concentration of the agent used were the same as those in Example 1 above. The difference was that the mycelial cake of wheat scab was inoculated at the base of the wheat seedling stem. Five days after the application, the length of the lesion at the base of the wheat seedling stem was measured, and the control effect (%) was calculated.

[0385] The experimental results are listed in Table 5.

[0386] Table 5

[0387]

[0388] As can be seen from the data listed in Table 5:

[0389] Some of the isoindoline ketone derivatives with different product numbers listed in Tables 1 and 2 also showed certain therapeutic and protective effects against wheat scab.

[0390] It should be noted that:

[0391] Although Table 5 only lists the test results of some of the numbered products in Tables 1 and 2, it can be seen from the actual test results that other isoindolinone derivatives with different numbers listed in Tables 1 and 2 also showed certain therapeutic and protective effects against wheat scab. For the sake of brevity, they are not described in more detail here.

[0392] Example 3: The control effect of isoindololinone derivatives with structures shown in Formula I, Formula II, Formula III, or Formula IV on wheat seedlings, taking wheat sheath blight pathogen as an example.

[0393] The experimental method and the concentration of the agent used were the same as those in Example 1 above. The difference was that the mycelium of wheat sheath blight was inoculated at the base of the wheat seedling stem. Five days after the application, the length of the lesion at the base of the wheat seedling stem was measured, and the control effect (%) was calculated.

[0394] The experimental results are listed in Table 6.

[0395] Table 6

[0396]

[0397] As can be seen from the data listed in Table 6:

[0398] Some of the isoindoline ketone derivatives with different product numbers listed in Tables 1 and 2 also have certain therapeutic and protective effects against wheat sheath blight.

[0399] It should be noted that:

[0400] Although Table 5 only lists the test results of some of the numbered products in Tables 1 and 2, it can be seen from the actual test results that other isoindolinone derivatives with different numbers listed in Tables 1 and 2 also showed certain therapeutic and protective effects against wheat sheath blight. For the sake of brevity, they are not described in more detail here.

[0401] In conclusion, it can be seen that:

[0402] First, this invention provides a novel series of isoindolinone derivatives with an amide bond as the core structure. These novel series of isoindolinone derivatives are synthesized from substituted or unsubstituted isoindolinone groups, amino acids, biogenic amines, etc.

[0403] Secondly, since the isoindolinone derivatives provided by this invention are prepared by reacting substituted or unsubstituted phthalic anhydride with amino acids to generate compounds with isoindolinone structural units, and then organically combining them with biogenic amines, the preparation method is relatively simple, the raw materials are readily available and the cost is low, it is easy to industrialize, the yield is high, the economy is good, and it has great value for promotion and application.

[0404] Furthermore, the isoindololinone derivatives provided by this invention not only have superior control effects on a variety of plant diseases, but also exhibit good biocompatibility and can promote autophagy in biological cells. Their innovative mechanism of action opens up new ideas and methods for the application of isoindololinone structural unit compounds in the agricultural field.

[0405] In the description process of the above instruction manual:

[0406] The terms “this embodiment,” “an embodiment of the present invention,” “as shown,” “further,” etc., are used to indicate that the specific features, structures, materials, or characteristics described in the embodiment or example are included in at least one embodiment or example of the present invention.

[0407] In this specification, the illustrative expressions of the above terms are not necessarily directed to the same embodiments or examples, and the specific features, structures, materials or characteristics described may be combined or combined in any suitable manner in one or more embodiments or examples;

[0408] Furthermore, without creating contradictions, those skilled in the art can combine or integrate the different embodiments or examples described in this specification, as well as the features of the different embodiments or examples.

[0409] Finally, it should be noted that:

[0410] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention. Non-essential improvements, adjustments or substitutions made by those skilled in the art based on the content of this specification are all within the scope of protection claimed by the present invention.

Claims

1. The application of an isoindolinone derivative in agricultural fungicides, characterized in that: The application involves using compounds having the structure shown in Formula I or Formula II for the prevention and control of gray mold in tomatoes, sclerotinia stem rot in rapeseed, scab in wheat, and sheath blight in wheat. in: The structure of the compound of formula I is: In the compound of formula II: R 1 It is any one of hydrogen, fluorine, chlorine, nitro, methoxy, methyl, or amino; R 2 It can be -CH2-, -(CH2)2-, or -(CH2). 10 -; R 3 It is a thiol group or -(CH2). n NH2, where n is 1 to 5.

2. The application as described in claim 1, characterized in that: The preparation method of the isoindolinone derivative includes steps a~b~c or a~b~d in the following synthetic route; in: Step a involves reacting substituted or unsubstituted phthalic anhydride with an amino acid to obtain an aminolysis product; Step b involves reacting the amino hydrolysate with thionyl chloride under reflux to form an acylated amino hydrolysate, i.e., intermediate A. Step c involves reacting intermediate A with putrescine to obtain an isoindololinone derivative having the structure shown in Formula I. Step d involves reacting intermediate A with cystamine to obtain an isoindolone derivative having the structure shown in Formula II.

3. The application as described in claim 2, characterized in that, Step a includes: Amine hydrolysis is carried out in glacial acetic acid with substituted or unsubstituted phthalic anhydride and an equimolar amount of amino acid. After the reaction is completed, water is added to form a mixed solution. The mixed solution is then sonicated to produce a solid precipitation. The solid is filtered to obtain a white solid. The solid is dried to obtain the aminohydrolysate.

4. The application as described in claim 3, characterized in that: The aminolysis reaction is carried out at a temperature of 100°C for 3 hours. The reaction is monitored by silica gel thin-layer chromatography. The aminolysis reaction ends when the silica gel thin-layer chromatographic spots of substituted or unsubstituted phthalic anhydrides in the reaction system disappear.

5. The application as described in claim 2, characterized in that, Step b includes: The aminohydrolysate was dissolved in thionyl chloride and then heated under reflux to undergo an acylation reaction to obtain a yellow oily intermediate A, wherein the acylation reaction lasted for 2 hours.

6. The application as described in claim 2, characterized in that, Step c includes: In putrescine, tetrahydrofuran was added as a solvent, and the acylamine chloride hydrolysate, intermediate A, was added dropwise under ice bath conditions for an ice bath reaction, followed by a room temperature reaction. After the reaction was completed, a room temperature reaction product was obtained. The room temperature reaction product was then purified by a pulping method to obtain a pale yellow powder solid containing the isoindoline ketone derivative with the structure shown in Formula I.

7. The application as described in claim 6, characterized in that: The ice bath reaction lasts for 5 minutes, and the room temperature reaction lasts for 2 hours. The room temperature reaction is tracked by silica gel thin-layer chromatography. The room temperature reaction ends when the silica gel thin-layer chromatographic spot of the ammonium chloride hydrolysate, i.e., intermediate A, disappears.

8. The application as described in claim 6, characterized in that: The purification method described above involves adding ethyl acetate to the reaction product at room temperature to obtain an ethyl acetate mixture, then stirring the ethyl acetate mixture in a high-speed magnetic stirrer, filtering to remove solid impurities, and then concentrating and drying the clarified liquid obtained from the filtration to obtain the purified isoindolone derivative having the structure shown in Formula I.

9. The application as described in claim 2, characterized in that, Step d includes: N,N-dimethylformamide was added as a solvent to a 4-dimethylaminopyridine solution of cystamine, and the acylamine chloride hydrolysate, intermediate A, was slowly added dropwise under ice bath conditions. The reaction was first carried out in an ice bath and then at room temperature. After the reaction was completed, a small amount of water was added to quench the reaction. The product was then extracted with ethyl acetate, the organic phase was collected, dried, and concentrated to obtain a deep yellow crude product. The crude product was then purified by silica gel column chromatography to obtain a yellow powdery solid, which is the isoindololinone derivative with the structure shown in Formula II.

10. The application as described in claim 9, characterized in that: The ice bath reaction lasts for 5 minutes, and the room temperature reaction lasts for 3 hours, both of which are carried out under nitrogen protection. The crude product is purified by silica gel column chromatography using a mixed solvent of dichloromethane and methanol in a ratio of 25:1.

Images