A morpholine-containing amide compound, a preparation method and application thereof

Abstract

The application belongs to the field of fruit and vegetable preservation and agricultural sterilization, and provides a morpholine-containing amide compound, a preparation method and application thereof. The method mainly utilizes N-(2-aminoethyl)morpholine and organic acids with different heterocyclic structures as raw materials, and through the catalysis of a condensing agent, an amidation reaction occurs in a dry organic solvent, thereby generating the morpholine-containing amide compound. The preparation process is simple, safe and environmentally friendly, the product yield is as high as 87%, and the product exhibits significant tyrosinase inhibition and good sterilization activity.

Description

Technical Field

[0001] This invention belongs to the field of fruit and vegetable preservation and agricultural sterilization, and specifically relates to an amide compound containing morpholine, its preparation method and application. Background Technology

[0002] Tyrosinase, also known as polyphenol oxidase, is widely found in plants and animals such as apples, mushrooms, and humans, as well as in microorganisms such as molds. Tyrosinase is a key rate-limiting enzyme in melanin biosynthesis. It catalyzes the hydroxylation of monophenols to ortho-diphenols, and further catalyzes the oxidation of ortho-diphenols to ortho-quinones, thereby forming melanin. Melanin causes browning in fruits, vegetables, and some aquatic products, thus reducing their shelf life and market value.

[0003] Tyrosinase inhibitors reduce melanin production by decreasing the catalytic efficiency of tyrosinase. Currently, tyrosinase inhibitors are widely available, including natural plant extracts, microbial sources, and chemical synthesis. Commonly used tyrosinase inhibitors include arbutin, vitamin C, ascorbic acid, hydroquinone, polyphenols, and aromatic alcohols. However, these additives suffer from problems such as low efficacy, poor stability, poor safety, and high cost in long-term use. Therefore, developing safe, non-toxic, stable, and highly active tyrosinase inhibitors is of great significance.

[0004] Morpholine rings contain nitrogen and oxygen atoms, making them readily chelated with Cu ions in tyrosinases. They also readily form hydrogen bonds in vivo, participating in physiological processes and exhibiting good biocompatibility. Morpholine compounds possess diverse biological activities, such as bactericidal, antioxidant, anti-browning, and antiviral properties, playing a crucial role in medicine, food, and agricultural sterilization. Summary of the Invention

[0005] To address the above-mentioned technical problems, this invention provides an amide compound containing morpholine. This compound has a simple structure, mild synthesis conditions, is easy to operate, and yields a high amount of product.

[0006] The technical solution of this invention is:

[0007] This invention provides an amide compound containing morpholine, with the following general structural formula:

[0008]

[0009] Wherein, R is any one of the following: pyrazole ring, pyrimidine ring, benzopyran ring and its derivatives;

[0010] Preferably, the general structural formula is:

[0011]

[0012] Among them, R 1 R2 , R 3 , R 4 is selected from any one of -H, -OH, -OR, -R, -CX3, -CHX2, -CH2X, -X.

[0013] The present application also provides a preparation method of the morpholine-containing amide compound, comprising the following steps:

[0014] The reagent I and the reagent II are dissolved in the organic solvent A, then a condensing agent is added, and the reaction is carried out at room temperature for 10-18 hours; after the reaction is completed, washing and drying are carried out, then extraction is carried out with the organic solvent C, and then the morpholine-containing amide compound is obtained after column chromatography purification, which is denoted as compound III;

[0015] The reagent I is a morpholine derivative;

[0016] The reagent II is an organic acid;

[0017] The organic solvent A is selected from any one of dichloromethane, 1,2-dichloroethane, diethyl ether, tetrahydrofuran, acetonitrile, and toluene;

[0018] The organic solvent C is selected from any one of dichloromethane, ethyl acetate, diethyl ether, and cyclohexane.

[0019] Preferably, the reagent I is N-(2-aminoethyl) morpholine;

[0020] The reagent II is selected from any one of 3-(difluoromethyl)-1H-pyrazole-4-carboxylic acid, 3-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid, 4-pyrimidine carboxylic acid, 2-cyclopropyl-6-hydroxy pyrimidine-4-carboxylic acid, 2-chloropyrimidine-4-carboxylic acid, and 2-oxo-2H-benzopyran-3-carboxylic acid, and is not limited to the above-mentioned types, but can also be other organic acids;

[0021] The condensing agent is composed of a condensing agent I and a condensing agent II, wherein the molar ratio of the condensing agent I to the condensing agent II is 1-3:1;

[0022] The condensing agent is selected from any one of the following: the condensing agent I is 4-dimethylaminopyridine, and the condensing agent II is dicyclohexyl carbodiimide;

[0023] Alternatively, the condensing agent I is dicyclohexyl carbodiimide, and the condensing agent II is 4-dimethylaminopyridine;

[0024] Alternatively, the condensing agent I is 1-hydroxybenzotriazole, and the condensing agent II is 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride;

[0025] Alternatively, the condensing agent I is 2-(7-azobenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and the condensing agent II is N,N-diisopropyl ethylamine.

[0026] The organic solvent C is selected from any one of dichloromethane, ethyl acetate, acetone, cyclohexane;

[0027] The developing agent used in column chromatography is selected from any two of methanol, ethyl acetate, dichloromethane, wherein the volume ratio of methanol: ethyl acetate is (1-3):(1-6); or the volume ratio of methanol:dichloromethane is 4:1.

[0028] Preferably, the molar ratio of reagent I to reagent II is 1:1.2-2.

[0029] Preferably, the molar ratio of reagent I: reagent II: condensing agent is 1:1:1.5-4; and the millimolar volume ratio of reagent II to organic solvent A is 1 mmol: 3-10 mL.

[0030] The application of the morpholine-containing amide compound in the preparation of fruit and vegetable fresh-keeping products and the application of the morpholine-containing amide compound in the preparation of agricultural fungicide products are also the focus of the present application.

[0031] The present application has the following advantages and effects relative to the prior art:

[0032] (1) The morpholine-containing amide compound provided by the present application has a simple structure;

[0033] (2) The morpholine-containing amide compound provided by the present application has a simple synthesis method, mild reaction conditions, high yield, and is easy to produce industrially;

[0034] (3) The morpholine-containing amide compound provided by the present application has good tyrosinase inhibitory activity and good application value in fruit and vegetable preservation;

[0035] (4) The morpholine-containing amide compound provided by the present application has good fungicidal activity and potential application value in agricultural fungicides. BRIEF DESCRIPTION OF DRAWINGS

[0036] Figure 1 is a preparation method flow chart;

[0037] Figure 2 is a molecular structure formula of compound IIIa-III f;

[0038] Figure 3 is the hydrogen spectrum of IIIa in Example 1 ( 1 H NMR, 400MHz, CDCl3);

[0039] Figure 4 is the carbon spectrum of IIIa in Example 1 ( 13 C NMR, 101MHz, CDCl3);

[0040] Figure 5 The mass spectra of IIIa in Example 1;

[0041] Figure 6 The proton spectrum of IIIb in Example 1 ( 1 H NMR, 400MHz, CDCl3);

[0042] Figure 7 The carbon spectrum of Ⅲb in Example 1 ( 13 C NMR, 101MHz, CDCl3);

[0043] Figure 8 The mass spectra of IIIb in Example 1;

[0044] Figure 9 The proton spectrum of IIIc in Example 1 ( 1 H NMR, 400MHz, CDCl3);

[0045] Figure 10 The carbon spectrum of IIIc in Example 1 ( 13 C NMR, 101MHz, CDCl3);

[0046] Figure 11 The mass spectra of IIIc in Example 1;

[0047] Figure 12 The proton spectrum of IIId in Example 1 ( 1 H NMR, 400MHz, DMSO);

[0048] Figure 13 The carbon spectrum of IIId in Example 1 ( 13 C NMR, 101MHz, DMSO);

[0049] Figure 14 The mass spectra of IIId in Example 1;

[0050] Figure 15 The proton spectrum of IIIe in Example 1 ( 1 H NMR, 400MHz, DMSO);

[0051] Figure 16 The carbon spectrum of Ⅲe in Example 1 ( 13 C NMR, 101MHz, DMSO);

[0052] Figure 17 The mass spectra of Ⅲe in Example 1;

[0053] Figure 18 The proton spectrum of IIIf in Example 1 ( 1 HNMR, 400MHz, CDCl3);

[0054] Figure 19 The carbon spectrum of Ⅲf in Example 1 is shown in the following table: 13 C NMR, 101 MHz, CDCl3);

[0055] Figure 20 The mass spectrum of Ⅲf in Example 1 is shown in the following table. DETAILED DESCRIPTION

[0056] In order to make the skilled in the art better understand the present application, the present application will be further described in conjunction with the specific embodiments.

[0057] The pharmaceuticals involved in the present application are purchased from Shanghai Maikelin Biochemical Technology Co., Ltd., Shanghai Annaiji Chemical Co., Ltd., Beijing Bailingwei Technology Co., Ltd., Shanghai Aladdin Biochem Technology Co., Ltd., Tianjin Kaitong Chemical Reagent Co., Ltd.

[0058] Preparation and characterization of the morpholine-containing amide compound in Example 1

[0059] (1) Reagents

[0060] Table 1 Reagent name and abbreviation

[0061]

[0062]

[0063] (2) Yield calculation method:

[0064] Yield = actual yield / theoretical yield x 100%

[0065] Preparation and characterization of the morpholine-containing amide compound in the present application Figure 2 Compounds Ⅲa-Ⅲf), and the product is characterized.

[0066] 1.1 Preparation and characterization of compound Ⅲa

[0067] As shown in the flow chart, Figure 1 In a 100 mL round-bottom flask, 3-(difluoromethyl)-1H-pyrazole-4-carboxylic acid 1.162 g (6.6 mmol) was taken, 0.859 g (6.6 mmol) of N-(2-aminoethyl) morpholine was added, 25 mL of dry dichloromethane, 1.64 g (8.56 mmol) of EDCI, 0.4 g (3.3 mmol) of DMAP were added in turn, and stirred at room temperature for 12 h. The reaction solution was washed with water, dried with anhydrous sodium sulfate, extracted with dichloromethane, and purified by column chromatography after desolvation. The developing agent for column chromatography was methanol: EA (volume ratio 3:1). A light orange solid was obtained, the yield was 61%, the melting range was 84-85℃.

[0068] and the data of the H NMR, C NMR and mass spectrum of compound IIIa are shown as follows: Figures 2 to 4

[0069] 1 H NMR (400 MHz, CDC13) δ (ppm) 7.89 (s, 1H, N-H), 6.85 (t, J = 54.1 Hz, 1H, Pyrazole-H), 3.87 (s, 3H, CH3), 3.67 (t, J = 4.7 Hz, 4H, Nam-CH2), 3.45 (q, J = 5.6 Hz, 2H, CH2), 2.82 (s, 1H, Nam-CH), 2.51 (t, J = 6.0 Hz, 2H, CH2), 2.45 (t, J = 4.7 Hz, 4H, Nam-CH2). 13 C NMR (101 MHz, CDC13) δ (ppm) 161.20, 142.94, 134.83, 116.75, 113.83, 111.51, 109.19, 66.82, 56.51, 53.14, 39.43, 35.92. IR (KBr) v cm-1: 3360 (N-H), 1636 (C=0), 1557 (C=N), 1271 (C-N); HRMS (ESI) calcd for [M+H] -1 : 289.1476, found 289.1480. + : 12 : 19 :

[0070] 1.2 Preparation and characterization of compound IIIb

[0071] Take 3-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid 6.6 mmol in a 100 mL round bottom flask, add N-(2-aminoethyl) morpholine 0.859 g (6.6 mmol) in turn 25 mL of dry dichloromethane, 1.64 g (8.56 mmol) EDCI, 0.4 g (3.3 mmol) DMAP, stirring at room temperature for 12 h. The reaction solution is washed with water, dried with anhydrous sodium sulfate, extracted with dichloromethane, and purified by column chromatography after desolvation. The developing agent of column chromatography is methanol: EA (volume ratio is 3:1). Orange solid, yield: 61%, melting point: 95-96°C;

[0072] and the data of the H NMR, C NMR and mass spectrum of compound IIIb are shown as follows: Figures 5 to 7

[0073] 1 ​​H NMR (400 MHz, CDC13) δ (ppm) 7.96 (s, 1H, N-H), 6.87 (s, 1H, Pyrazole-H), 3.96 (s, 3H, CH3), 3.72 (d, J = 4.6 Hz, 4H, Nam-CH2), 3.51 (s, 2H, CH2), 2.56 (t, J = 5.9 Hz, 2H, CH2), 2.49 (t, J = 4.6 Hz, 4H, Nam-CH2). 13 C NMR (101 MHz, CDC13) δ (ppm) 160.32, 135.73, 122.39, 119.72, 117.46, 66.86 (2C), 56.30, 53.13 (2C), 39.75, 35.94. IR (KBr) v cm -1 : 3242 (N-H), 1639 (C=0), 1573 (C=N), 1232 (C-N); HRMS (ESI) calcd for [M+H] + : 307.1382, found 307.1387. 12 H 18 N4O2F3: 307.1382, found 307.1387.

[0074] 1.3 Preparation and characterization of compound IIIc

[0075] Take 4-pyrimidine carboxylic acid 3 mmol in 100 mL round bottom flask, add N-(2-aminoethyl) morpholine 0.391 g (3 mmol) in turn 25 mL dry dichloromethane, 0.958 g (5 mmol) EDCI, 0.611 g (5 mmol) DMAP, stirring at room temperature for 16 h. The reaction solution is washed with water, dried with anhydrous sodium sulfate, extracted with dichloromethane, and purified by column chromatography after desolvation. The developing agent of column chromatography is methanol: EA (volume ratio is 1:1). Yellow solid, yield: 55.4%; melting point: 89-90 °C.

[0076] The nuclear magnetic hydrogen spectrum and nuclear magnetic carbon spectrum mass spectrum data of compound IIIc are as follows:

[0077] 1 H NMR (400 MHz, DMSO-d6) δ (ppm) 9.33 (s, 1H, Pyrim-H), 9.06 (s, 1H, Pyrim-H), 8.92 (d, J = 5.9 Hz, 1H, N-H), 8.03 (dd, J = 5.1, 1.4 Hz, 1H, Pyrim-H), 3.57 (t, J = 4.6 Hz, 4H, Nam-CH2), 3.47 (m, 2H, CH2), 2.51 (m, 2H, CH2), 2.41 (t, J = 4.6 Hz, 4H, Nam-CH2).13 C NMR (151 MHz, DMSO-d6) δ (ppm) 162.83, 160.14, 158.33, 156.82, 118.90, 66.67 (2C), 57.40, 53.63 (2C), 36.52. IR (KBr) v cm"1: 3242 (N-H), 1639 (C=O), 1573 (C=N), 1232 (C-N); HRMS (ESI) calcd for [M+H] -1 :3242(N-H),1639(C=O),1573(C=N),1232(C-N);HRMS(ESI)calcd for[M+H] + :C 11 H 16 N4O2:237.1352,found 237.1355.

[0078] 1.4 Preparation and characterization of compound III d

[0079] Into a 100 mL round bottom flask, 2-cyclopropyl-6-hydroxy pyrimidine-4-carboxylic acid 6.6 mmol was added, followed by N-(2-aminoethyl)morpholine 0.859 g (6.6 mmol), 25 mL dry dichloromethane, 1.64 g (8.56 mmol) EDCI, 0.4 g (3.3 mmol) DMAP. The reaction solution was stirred at room temperature for 18 h. The reaction solution was washed with water, dried over anhydrous sodium sulfate, extracted with dichloromethane, and purified by column chromatography after desolvation. The developing agent for column chromatography was methanol: EA (volume ratio 3: 1). Light yellow solid, yield 67.3%; melting point 195-196 °C;

[0080] The nuclear magnetic hydrogen spectrum, nuclear magnetic carbon spectrum and mass spectrum data of compound III d are as follows:

[0081] 1 H NMR (400 MHz, DMSO-d6) δ (ppm) 12.87 (s, 1H, OH) 8.48 (s, 1H, NH), 6.56 (s, 1H, Pyrim-H), 3.64 (d, J = 26.4 Hz, 4H, Nam-CH2), 3.33 (s, 2H, CH2), 2.50 (m, 6H, Nam-CH2), 1.97 (s, 1H, Cpr-CH), 1.49 (m, 4H, Cpr-CH2); 13 CNMR (101 MHz, DMSO) δ (ppm) 165.19, 163.34, 162.63, 154.81, 109.72, 66.81 (2C), 56.92, 53.48 (2C), 39.35, 36.10, 13.92, 10.67. IR (KBr) v cm"1: 3366 (N-H), 1658 (C=O), -1 :3366(N-H),1658(C=O),

[0082] 1509 (C=N), 1113 (C-N); HRMS (ESI) calcd for [M+H] + : C 14 H 21 N4O3: 293.1614; found 293.1618.

[0083] 1.5 Preparation and characterization of compound Ⅲe

[0084] Take 2-chloropyrimidine-4-carboxylic acid 3 mmol in a 100 mL round-bottom flask, add N-(2-aminoethyl) morpholine 0.391 g (3 mmol) in turn 25 mL of dry dichloromethane, 0.611 g (5 mmol) EDCI, 0.611 g (5 mmol) DMAP, stirring at room temperature for 14 h. The reaction solution is washed with water, dried with anhydrous sodium sulfate, extracted with dichloromethane, and purified by column chromatography after desolvation. The developing agent of column chromatography is methanol: EA (volume ratio is 1:6). Light yellow solid, yield 51.8%; melting point 86-87℃;

[0085] And the data of the nuclear magnetic hydrogen spectrum, nuclear magnetic carbon spectrum and mass spectrum of compound Ⅲe are as follows:

[0086] 1 H NMR (400 MHz, DMSO-d6) δ (ppm) 9.01 (s, 1H, Pyrim-H), 8.92 (s, 1H, NH), 8.02 (d, J = 6.5 Hz, 1H, Pyrim-H), 3.56 (s, 4H, Nam-CH2), 3.18 (s, 2H, CH2), 2.49 (s, 4H, Nam-CH2), 2.42 (s, 2H, CH2). 13 C NMR (101 MHz, DMSO-d6) δ (ppm) 163.42, 161.67, 160.68, 160.34, 118.21, 66.73, 66.61, 57.31, 57.12, 53.59, 53.51. HRMS (ESI) calcd for [M+H] + : C 11 H 15 ClN4O2: 291.0962; found 291.0965.

[0087] 1.6 Preparation and characterization of compound Ⅲf

[0088] Into a 100 mL round bottom flask, 2-oxo-2H-chromen-3-carboxylic acid 6 mmol was taken, to this N-(2-aminoethyl)morpholine 0.781 g (6 mmol) was added. To this 25 mL of dry dichloromethane, 1.917 g (10 mmol) of EDCI, 0.773 g (6 mmol) of DMAP was added successively and stirred for 18 h at room temperature. The reaction solution was washed with water, dried over anhydrous sodium sulphate, extracted with ethyl acetate and purified by column chromatography after desolventizing. The eluent used for column chromatography was methanol:EA (1:1 by volume). Pale yellow solid, yield 87.3%; melting range 147-148°C;

[0089] The data of1H NMR,13C NMR and mass spectrum of compound III f are as follows:

[0090] 1 H NMR (400 MHz, CDC13) δ (ppm) 9.01 (s, 1H, Ccr-1H), 8.90 (s, 1H, NH), 7.74-7.65 (m, 2H, Ph-2H), 7.39 (dd, J = 12.2, 7.9 Hz, 2H, Ph-2H), 3.77-3.74 (m, 4H, Nam-CH2), 3.59 (q, J = 6.0 Hz, 2H, CH2), 2.61 (t, J = 6.3 Hz, 2H, CH2), 2.53 (t, J = 4.6 Hz, 4H, Nam-CH2). 13 C NMR (101 MHz, CDC13) δ (ppm) 161.46, 161.28, 154.46, 148.13, 134.00, 129.80, 125.26, 118.67, 118.64, 116.63, 67.07 (2C), 56.81 (2C), 53.43, 36.66. IR (KBr) v cm-1: 3323 (N-H), 1689 (C=0), 1540 (C=N), 1240 (C-N); HRMS (ESI) calcd for [M+H] -1 : 291.0962; found 303.1350. + : 291.0962; found 303.1350. 16 : 291.0962; found 303.1350. 18 : 291.0962; found 303.1350.

[0091] Example 2

[0092] 2.1 Tyrosinase inhibitory activity experiment

[0093] The compounds IIIa-IIIf in Example 1 were dissolved in DMSO respectively, and diluted with PBS to prepare solutions with different concentrations (the concentrations of the solutions are shown in Table 1). 20 μL of the solution, 30 μL of tyrosinase (200 U / mL) and 100 μL of phosphate buffer (50 mmol / L, pH = 6.8) were added to each well of a 96-well plate. After incubation at 25°C for 20 min, 50 μL of L-DOPA (2 mmol / L) was added as a substrate. The solution was mixed thoroughly, and then incubated at 25°C for 10 min. Finally, the absorbance was measured at 475 nm using a microplate reader. DMSO was used as a blank control, and kojic acid was used as a positive control. The experiment was repeated three times. The inhibition rate was calculated according to the following formula:

[0094] Inhibition rate (%) = (1 - ΔAsample / ΔAcontrol) x 100%,

[0095] wherein ΔAsample is the absorbance of the sample containing the target compound;

[0096] ΔAcontrol is the absorbance of the DMSO control.

[0097] The results of the activity test are shown in Table 1.

[0098] Table 2 Tyrosinase inhibition activity of the target compounds IIIa-IIIf

[0099]

[0100] The results show that the compounds in the series all exhibit good tyrosinase inhibition activity, and have a concentration inhibition effect, and can be used for fruit and vegetable preservation, and for the production and preparation of related products.

[0101] 2.2 Bactericidal activity

[0102] Experimental method: Each of the test compounds IIIa-IIIf in Example 1 was dissolved in DMSO to prepare a 1% Tween 80-DMSO stock solution for use. The mycelial growth method was used to evaluate the indoor bactericidal activity of the test compounds IIIa-IIIf at doses of 50 mg / L and 100 mg / L on the test targets, with DMSO as a control.

[0103] The bacteriostatic rate was calculated according to the following formula:

[0104]

[0105] The results of the activity test are shown in Table 2.

[0106] Table 3 Bactericidal activity of the compounds IIIa-IIIf

[0107]

[0108] Experiments show that the six compounds IIIa-IIIf all show certain fungicidal activity, for example, at 50 mg / L, the inhibition rates of the compounds IIIa, IIIe and IIIf on Sclerotinia sclerotiorum, Botrytis cinerea, Penicillium citrinum and Sclerotinia homoeocarpa are all above 50%; at 100 mg / L, the inhibition rates of IIIa, IIIe and IIIf on Sclerotinia sclerotiorum are above 70%, the inhibition rates on Botrytis cinerea are about 60%, the inhibition rates on Penicillium citrinum are all above 50%, and the inhibition rates on Sclerotinia homoeocarpa are about 60%, which can be used to prepare fungicides and other products.

[0109] Examples 3-6

[0110] The compound IIIa is prepared according to the reagents in the following table, and other reagents and dosages in the preparation process are the same as the preparation steps of the compound IIIa in Example 1.

[0111] Table 4 Reagents participating in the reaction in Examples 3-6

[0112]

[0113] In summary, the present application provides a morpholine-containing amide compound and a preparation method thereof. The method mainly uses N-(2-aminoethyl) morpholine and organic acids with different heterocyclic structures as raw materials, and through the catalysis of a condensing agent, an amide reaction occurs in a dry organic solvent, thereby generating a morpholine-containing amide compound. The preparation process is simple, safe and environmentally friendly, the product yield is high up to 87%, and the product shows significant tyrosinase inhibition effect and good fungicidal activity.

[0114] The above is only the preferred embodiment of the present application, and does not limit the patent scope of the present application, and any equivalent changes and modifications made within the scope of the present application shall still fall within the scope of the present application.

Claims

1. The application of morpholine-containing amide compounds in the preparation of fruit and vegetable preservation products and / or agricultural fungicide products, characterized in that, The structure of the morpholine-containing amide compound is as follows: 。 2. The application as described in claim 1, characterized in that, The preparation process of the IIId is as follows: 6.6 mmol of 2-cyclopropyl-6-hydroxypyrimidine-4-carboxylic acid was placed in a 100 mL round-bottom flask, and 0.859 g of N-(2-aminoethyl)morpholine was added. 25 mL of dry dichloromethane, 1.64 g of EDCI, and 0.4 g of DMAP were added sequentially. The mixture was stirred at room temperature for 18 h. The reaction solution was washed with water, dried over anhydrous sodium sulfate, extracted with dichloromethane, and purified by column chromatography after solvent removal. The developing solvent for column chromatography was methanol:EA, with a volume ratio of 3:

1. The preparation process of the Ⅲe is as follows: Take 3 mmol of 2-chloropyrimidine-4-carboxylic acid into a 100 mL round-bottom flask, add 0.391 g of N-(2-aminoethyl)morpholine, and then add 25 mL of dry dichloromethane, 0.611 g of EDCI, and 0.611 g of DMAP. Stir at room temperature for 14 h. Wash the reaction solution with water, dry it with anhydrous sodium sulfate, extract it with dichloromethane, and purify it by column chromatography after solvent removal. The developing solvent for column chromatography is methanol:EA, with a volume ratio of methanol:EA of 1:6.

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