Synthesis and application of a class of phenylacetamide derivatives containing alpha-piperazinyl

Abstract

The application discloses a kind of alpha-piperazinyl containing phenylacetamide derivative, it is characterized in that: its structural formula is as follows: wherein R1 For hydrogen, trifluoromethyl, methoxy or halogen, R2 For hydrogen, acetamidyl, methyl or halogen, R3 For methyl or acetyl.The application designs, synthesizes a series of alpha-piperazinyl containing novel phenylacetamide derivative, and biological activity test finds that the compound not only shows excellent bacteriostatic activity to brown rot pathogen of stone fruit, but also can effectively inhibit spore germination of brown rot pathogen of stone fruit;In addition, it has good protection and prevention and treatment effect to brown rot of picked pear.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, and in particular to a method for preparing a class of phenylacetamide derivatives containing α-piperazine groups and their application in the prevention and control of fungal diseases in plants and in the prevention and control of fungal diseases in fruits after harvesting. Background Technology

[0002] In agricultural production, pesticides are an indispensable component, making a significant contribution to agricultural development and ensuring increased food yields. Many phenylacetamide compounds have been developed for pesticide production. For example, the herbicide bismuth subtilis, introduced in 1989, is mainly used to control annual grasses or certain broadleaf weeds, suitable for ornamental plants such as peanuts, peppers, tomatoes, cotton, and soybeans. Additionally, Syngenta launched a novel fungicide, α-hydroxylated phenylacetamide-mandipropamid, in 2008. Its primary target is oomycete diseases. This fungicide effectively prevents the germination of pathogenic spores, thereby inhibiting mycelial growth and spore formation. Simultaneously, it rapidly adsorbs onto the waxy layer of plant surfaces, providing the most durable protective barrier for fungal disease control. However, the improper and abusive use of pesticides has led to pesticide resistance and pollution problems. There is an urgent need to research and develop more agrochemicals with novel structures, high activity, high selectivity and environmental protection properties, so as to provide a guarantee for the healthy and sustainable development of agricultural production.

[0003] Piperazine rings, as key six-membered heterocyclic compounds, are characterized by low toxicity, ease of forming multiple hydrogen or ionic bonds, and the ability to regulate the lipid-water partition coefficient and pH stability of drugs. They are considered bridging molecules that can effectively connect highly active groups, thereby enhancing biological activity, and their chemical properties can be optimized by introducing different groups. N-substituted piperazine compounds have been reported to possess a wide range of pharmacological activities, such as antibacterial, anticancer, and antituberculosis activities. However, reports on piperazine-containing compounds in pesticide development are relatively few. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to provide a class of phenylacetamide derivatives containing α-piperazine group for the prevention and control of fungal diseases in plants and for the prevention and control of fungal diseases in fruits after harvesting.

[0005] The technical solution of this invention: a class of phenylacetamide derivatives containing α-piperazine groups, the structural formula of which is as follows: R1 is hydrogen, trifluoromethyl, methoxy, or halogen; R2 is hydrogen, acetamido, methyl, or halogen; and R3 is methyl or acetyl.

[0006] Preferably, the substitution position of R1 is a single substitution at the ortho, meta, or para position, and the substituent of the phenyl group of R2 is hydrogen, acetamido, methyl, or halogen.

[0007] The R2-substituted phenyl group is a single or disubstituted group at the ortho, meta, or para position.

[0008] A method for preparing a class of phenylacetamide derivatives containing an α-piperazine group, the reaction formula is as follows:

[0009]

[0010] The preparation method of the phenylacetamide derivative containing α-piperazine group and its application in the prevention and control of fungal diseases in plants and post-harvest fruits.

[0011] The beneficial effects of this invention are as follows: This invention constructs a novel active skeleton using mandelic acid as a starting material. First, an amide structure is constructed by combining it with a substituted amino group. Then, an active "piperazine" structure is introduced after halogenation of the hydroxyl group of mandelic acid. Finally, a series of phenylacetamide derivatives containing α-piperazine groups are designed and synthesized. Highly active compounds are sought by adjusting the structure of three parts of the active skeleton (the benzene ring based on mandelic acid, the formamide-containing compound, and the piperazine moiety). Compared with traditional commercially available amide structures, this invention constructs a novel active phenylacetamide skeleton containing α-piperazine groups using mandelic acid as a starting material, instead of synthesizing thioether compounds through the introduction of "oxadiazole" or "thiadiazole" structures. Furthermore, compared with commercially available structures starting from mandelic acid, this invention does not obtain a series of thioether compounds by substituting hydrogen atoms at the "hydroxyl" group. Instead, it introduces the active unit "piperazine" at this position by halogenating the "hydroxyl" group in the mandelic acid structure. Highly active compounds are discovered by exploring the influence of different structural units in the active skeleton on activity. In vitro experiments revealed that these compounds exhibit good antifungal activity against *Brachys spp.*, the causal agent of brown rot in stone fruits. They also effectively inhibit the germination of *Brachys spp.* Furthermore, they provide good protection and control against brown rot in post-harvest pears. These compounds possess novel structures and wide applications, providing an important scientific basis for the research and development of new pesticides. Attached Figure Description

[0012] Figure 1 The effects of the compound and the commercially available agent picoxystrobin on the germination and morphology of spores of the brown rot fungus *Monilinia fructicola* were investigated.

[0013] Figure 2The compound and the commercially available agent picoxystrobin under different treatments showed good protective and control effects against brown rot in pears after harvest (2A, therapeutic effect; 2B, protective effect). Detailed Implementation

[0014] Synthetic routes for pyrazole hydrazone target compounds:

[0015]

[0016] Starting with amygdalin and substituted aniline, the target compound Z was synthesized through condensation, halogenation, and substitution.

[0017] Preparation of intermediates

[0018] Preparation of substituted phenyl-2-hydroxy-2-acetic acid aryl ester (B)

[0019]

[0020] A mixed solution of substituted mandelic acid (0.8 g), substituted aniline (0.5 g), dichloromethane (10 mL), and 4-dimethylaminopyridine (0.2 g) was reacted at room temperature for 2 h. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.7 g) was added in an ice bath, and the reaction solvent was stirred for 6 h. The reaction was monitored by thin-layer chromatography. After the solvent was dried under vacuum by rotary evaporation, it was washed with ethyl acetate and water. The organic phase was retained, dried overnight with anhydrous sodium sulfate, and purified by silica gel column chromatography (dichloromethane / methanol = 5 / 1 to 3 / 1) to give intermediate B. The physicochemical and spectroscopic data are as follows:

[0021] N-(4-acetamidophenyl)-2-hydroxy-2-phenylacetamide (B1): white solid, yield 72%, melting point 223–225 °C. 1 HNMR(400MHz,DMSO-d6)δ9.87(s,1H,NH),9.85(s,1H,NH),7.61–7.59(m,2H,phenylH),7.52–7.47(m,4H,phenyl H),7.37–7.26(m,2H,phenyl H),7.31–7.27(m,1H,phenylH),6.40–6.39(d,J=4.0Hz,1H,OH),5.09–5.08(d,J=4.0Hz,1H,CH),2.01(s,3H,Ar-NHCOCH3); 13C NMR (101MHz, DMSO-d6) δ170.79,167.95,140.90,135.07,133.71,128.04,127.54,126.54,120.04,119.18,73.92,23.87.

[0022]

[0023] N-(4-acetamidophenyl)-2-hydroxy-2-(4-(trifluoromethyl)phenyl)acetamide (B2): white solid, yield 65%, melting point 227–229 °C. 1 H NMR(500MHz,DMSO-d6)δ9.98(s,1H,NH),9.90(s,1H,NH),7.74(s,4H,phenyl H),7.61–7.57(m,2H,phenyl H),7.50–7.46(m,2H,phenyl H),6.68(s,1H,OH),5.22(s,1H,CH),2.01(s,3H,Ar-NHCOCH3).

[0024]

[0025] N-(4-acetamidophenyl)-2-(3-chlorophenyl)-2-hydroxyacetamide (B3): white solid, yield 70%, melting point 233–235 °C. 1 H NMR(500MHz,DMSO-d6)δ9.92(s,1H,NH),9.89(s,1H,NH),7.59–7.56(m,3H,phenyl H),7.49–7.45(m,3H,phenyl H),7.40–7.34(m,2H,phenyl H),6.59–6.58(d,J=5.0Hz,1H,OH),5.12–5.11(d,J=5.0Hz,1H,CH),2.00(s,3H,Ar-NHCOCH3),

[0026]

[0027] N-(4-acetamidophenyl)-2-(3-fluorophenyl)-2-hydroxyacetamide (B4): white solid, yield 74%, melting point 221–223 °C. 1H NMR(500MHz,DMSO-d6)δ9.87(s,1H,NH),9.86(s,1H,NH),7.59–7.57(m,2H,phenyl H),7.49–7.47(m,2H,phenyl H),7.42–7.34(m,3H,phenyl H),7.14–7.10(m,1H,phenyl H),6.55–6.54(d,J=5.0Hz,1H,OH),5.12–5.11(d,J=5.0Hz,1H,CH),2.01(s,3H,Ar-NHCOCH3),

[0028]

[0029] N-(4-acetamidophenyl)-2-(4-bromophenyl)-2-hydroxyacetamide (B5): white solid, yield 74%, melting point 231–233 °C. 1 H NMR(500MHz,DMSO-d6)δ9.90(s,1H,NH),9.89(s,1H,NH),7.59–7.54(m,4H,phenyl H),7.49–7.45(m,4H,phenyl H),6.54–6.53(d,J=5.0Hz,1H,OH),5.08–5.07(d,J=5.0Hz,1H,CH),2.01(s,3H,Ar-NHCOCH3),

[0030]

[0031] N-(4-acetamidophenyl)-2-(4-fluorophenyl)-2-hydroxyacetamide (B6): white solid, yield 71%, melting point 226–228 °C. 1 H NMR(500MHz,DMSO-d6)δ9.89(s,2H,NH-NH),7.61–7.45(m,6H,phenyl H),7.20–7.16(m,2H,phenyl H),6.48–6.47(d,J=5.0Hz,1H,OH),5.10–5.07(d,J=10.0Hz,1H,CH),2.00(s,3H,Ar-NHCOCH3),

[0032]

[0033] N-(4-acetamidophenyl)-2-(4-chlorophenyl)-2-hydroxyacetamide (B7): white solid, yield 66%, melting point 224–226 °C. 1H NMR(500MHz,DMSO-d6)δ9.89(s,2H,NH-NH),7.61–7.45(m,6H,phenyl H),7.20–7.16(m,2H,phenyl H), 6.48–6.47 (d, J=5.0Hz, 1H, OH), 5.10–5.07 (d, J=10.0Hz, 1H, CH), 2.00 (s, 3H, Ar-NHCOCH3).

[0034]

[0035] N-(4-acetamidophenyl)-2-(4-methoxyphenyl)-2-hydroxyacetamide (B8): white solid, yield 75%, melting point 223–225 °C. 1 H NMR(500MHz,DMSO-d6)δ9.90(s,1H,NH),9.83(s,1H,NH),7.60–7.58(m,2H,phenyl H),7.49–7.47(m,2H,phenyl H),7.42–7.40(m,2H,phenyl H),6.92–6.89(m,2H,phenyl H),6.29(s,1H,OH),5.02(s,1H,CH),3.73(s,3H,OCH3),2.00(s,3H,Ar-NHCOCH3).

[0036]

[0037] N-(4-acetamidophenyl)-2-(2-chlorophenyl)-2-hydroxyacetamide (B9): white solid, yield 72%, melting point 221–223 °C. 1 H NMR(500MHz,DMSO-d6)δ9.97(s,1H,NH),9.89(s,1H,NH),7.63–7.60(m,2H,phenyl H),7.57–7.55(m,1H,phenyl H),7.50–7.48(m,2H,phenyl H),7.45–7.43(m,1H,phenyl H),7.38–7.31(m,2H,phenyl H),6.64–6.62(d,J=10.0Hz,1H,OH),5.46–5.45(d,J=5.0Hz,1H,CH),2.01(s,3H,Ar-NHCOCH3).

[0038]

[0039] 2-Hydroxy-N,2-Diphenylacetamide (B10): White solid, yield 64%, melting point 193–195 °C. 1 H NMR(500MHz,DMSO-d6)δ9.95(s,1H,NH),7.76–7.54(d,J=5.0Hz,2H,phenyl H),7.44–7.42(m,2H,phenyl H),7.38–7.(m,5H,phenyl H),7.20–7.16(m,1H,phenyl H),6.48(s,1H,OH),5.11(s,1H,CH),

[0040]

[0041] 2-Hydroxy-N-phenyl-2-(4-(trifluoromethyl)phenyl)acetamide (B11): white solid, yield 64%, melting point 191–193 °C. 1 H NMR (500MHz, CDCl3) δ10.05(s,1H,NH),7.75–7.74(d,J=5.0Hz,4H,phenylH),7.70–7.68(m,2H,phenyl H),7.30–7.27(m,2H,phenyl H),7.07–7.04(m,1H,phenylH),6.71–6.70(d,J=5.0Hz,1H,OH),5.25–5.24(d,J=5.0Hz,1H,CH).

[0042]

[0043] 2-(3-Chlorophenyl)-2-hydroxy-N-phenylacetamide (B12): White solid, yield 60%, melting point 191–193 °C. 1 H NMR(500MHz,CDCl3)δ9.98(s,1H,NH),7.70–7.67(m,2H,phenyl H),7.59–7.58(m,1H,phenyl H),7.49–7.47(m,1H,phenyl H),7.41–7.35(m,2H,phenyl H),7.31–7.27(m,2H,phenyl H),7.07–7.04(m,1H,phenyl H),6.62–6.61(d,J=5.0Hz,1H,OH),5.14–5.13(d,J=5.0Hz,1H,CH).

[0044]

[0045] 2-(3-Fluorophenyl)-2-hydroxy-N-phenylacetamide (B13): White solid, yield 65%, melting point 193–195 °C. 1 H NMR(500MHz,CDCl3)δ8.19(s,1H,NH),7.53–7.51(m,2H,phenyl H),7.38–7.29(m,4H,phenyl H),7.23–7.21(m,1H,phenyl H),7.15–7.12(m,1H,phenyl H),7.07–7.03(m,1H,phenyl H),7.20–7.19(d,J=5.0Hz,1H,OH),3.50(s,1H,CH).

[0046]

[0047] 2-(4-Bromophenyl)-2-hydroxy-N-phenylacetamide (B14): white solid, yield 69%, melting point 196–198 °C. 1 H NMR(500MHz,CDCl3)δ9.95(s,1H,NH),7.69–7.66(m,2H,phenyl H),7.57–7.54(m,2H,phenyl H),7.49–7.46(m,2H,phenyl H),7.30–7.26(m,2H,phenyl H),7.06–7.03(m,1H,phenyl H),6.57–6.56(d,J=5.0Hz,1H,OH),5.11–5.10(d,J=5.0Hz,1H,CH),

[0048]

[0049] 2-(4-Fluorophenyl)-2-hydroxy-N-phenylacetamide (B15): White solid, yield 65%, melting point 196–198 °C. 1 H NMR(500MHz,DMSO-d6)δ10.08(s,1H,NH),7.71–7.67(m,2H,phenyl H),7.56–7.53(m,2H,phenyl H),7.43–7.41(m,2H,phenyl H),7.31–7.26(m,2H,phenyl H),7.06–7.03(m,1H,phenyl H),6.64(s,1H,OH),5.15(s,1H,CH).

[0050]

[0051] 2-(4-Chlorophenyl)-2-hydroxy-N-phenylacetamide (B16): White solid, yield 67%, melting point 197–199 °C. 1 H NMR(500MHz,DMSO-d6)δ9.96(s,1H,NH),7.70–7.67(m,2H,phenyl H),7.57–7.55(m,2H,phenyl H),7.49–7.47(m,2H,phenyl H),7.30–7.27(m,2H,phenyl H),7.07–7.03(m,1H,phenyl H),6.57–6.56(d,J=5.0Hz,1H,OH),5.11–5.10(d,J=5.0Hz,1H,CH).

[0052]

[0053] 2-Hydroxy-2-(4-methoxyphenyl)-N-phenylacetamide (B17): White solid, yield 65%, melting point 192–194 °C. 1 HNMR(500MHz,CDCl3)δ9.68(s,1H,NH),7.58–7.55(m,2H,phenyl H),7.34–7.29(m,4H,phenyl H),7.09–7.06(m,1H,phenyl H),6.84–6.85(m,2H,phenyl H),5.63–5.62(d,J=5.0Hz,1H,OH),5.10–5.48(m,1H,CH),3.77(s,3H,Ar-OCH3).

[0054]

[0055] 2-(2-Chlorophenyl)-2-hydroxy-N-phenylacetamide (B18): White solid, yield 71%, melting point 194–196 °C. 1 H NMR(500MHz,DMSO-d6)δ10.09(s,1H,NH),7.73–7.70(m,2H,phenyl H),7.58–7.56(m,2H,phenyl H),7.45–7.43(m,1H,phenyl H),7.36–7.33(m,2H,phenyl H),7.32–7.29(m,2H,phenyl H),7.08–7.05(m,1H,phenyl H),6.77(s,1H,OH),5.48(s,1H,CH).

[0056]

[0057] N,2-Di(4-bromophenyl)-2-hydroxyphenylacetamide (B19): White solid, 74% yield, melting point 204–206 °C. 1 H NMR(500MHz,DMSO-d6)δ10.29(s,1H,NH),7.70–7.66(m,2H,phenyl H),7.57–7.54(m,2H,phenyl H),7.49–7.45(m,4H,phenyl H),6.61–6.60(d,J=5.0Hz,1H,OH),5.11–5.10(d,J=5.0Hz,1H,CH).

[0058]

[0059] 2-(4-Bromophenyl)-N-(4-chloro-2-tolyl)-2-hydroxyacetamide (B20): white solid, yield 77%, melting point 200–202 °C. 1 H NMR(500MHz, CDCl3)δ9.38(s,1H,NH),7.73–7.71(d,J=5.0Hz,1H,phenylH),7.59–7.57(m,2H,phenyl H),7.37–7.35(m,2H,phenyl H),7.18–7.16(m,2H,phenylH),5.48–5.46(m,1H,OH),5.25–5.24(d,J=10.0Hz,1H,CH),2.25(s,3H,Ar-CH3).

[0060]

[0061] 2-(4-Bromophenyl)-N-(2,4-Difluorophenyl)-2-hydroxyacetamide (B21): White solid, yield 74%, melting point 206-208℃. 1 H NMR(500MHz,CDCl3)δ9.60(s,1H,NH),8.04–8.00(m,1H,phenyl H),7.59–7.57(m,2H,phenyl H),7.35–7.33(m,2H,phenyl H),7.05–7.01(m,1H,phenyl H),6.96–6.92(m,1H,phenyl H),5.49–5.47(m,1H,OH),5.29–5.27(d,J=10.0Hz,1H,CH).

[0062]

[0063] 2-(4-Bromophenyl)-N-(4-Fluorophenyl)-2-hydroxyacetamide (B22): White solid, yield 74%, melting point 201–203 °C. 1 HNMR(500MHz,CDCl3)δ8.58(s,1H,NH),8.27–8.22(m,1H,phenyl H),7.54–7.49(m,2H,phenyl H),7.39–7.36(m,2H,phenyl H),7.14–7.04(m,3H,phenyl H),5.19(s,1H,OH),3.56(s,1H,CH).

[0064]

[0065] 2-(4-Bromophenyl)-N-(3-Fluorophenyl)-2-hydroxyacetamide (B23): White solid, yield 68%, melting point 207–209 °C. 1 H NMR(500MHz,CDCl3)δ10.18(s,1H,NH),7.68–7.64(m,1H,phenyl H),7.58–7.55(m,2H,phenyl H),7.52–7.49(m,1H,phenyl H),7.47–7.45(m,2H,phenyl H),7.35–7.19(m,1H,phenyl H),6.91–6.86(m,1H,phenyl H),6.64–6.62(d,J=10.0Hz,1H,OH),5.12–5.11(d,J=5.0Hz,1H,CH).

[0066] Preparation of substituted phenyl-2-chloro-2-acetic acid aryl esters (C)

[0067]

[0068] At room temperature, N,N-dimethylformamide (0.5 mL) was reacted with cyanuric chloride (TCT, 0.3 g) until fully reacted. Then, dichloromethane (20 mL) and substituted phenyl-2-hydroxy-2-acetic acid aryl ester (1 g) were added. After reacting for 3 hours at room temperature, the reaction was detected by thin-layer chromatography. Subsequently, ammonium chloride (NH4Cl) solution was added for 5-6 extractions, the organic phase was retained, and dried using anhydrous sodium sulfate. Finally, the intermediate C was purified by silica gel column chromatography (elution: petroleum ether / ethyl acetate = 5 / 1 to 2 / 1). The physicochemical and spectroscopic data are as follows:

[0069]

[0070] N-(4-acetamidophenyl)-2-chloro-2-phenylacetamide (C1): white solid, yield 47%, melting point 207–209 °C. 1 H NMR(500MHz,DMSO-d6)δ10.47(s,1H,NH),9.95(s,1H,NH),7.60–7.58(m,2H,phenylH),7.54–7.49(m,4H,phenyl H),7.44–7.38(m,3H,phenyl H),5.72(s,1H,CH),2.01(s,3H,Ar-NHCO CH3 ); 13 C NMR (101MHz, DMSO-d6) δ170.79,167.95,140.90,135.07,128.04,127.54,126.54,120.04,119.18,73.92,23.87.

[0071]

[0072] N-(4-acetamidophenyl)-2-chloro-2-(4-(trifluoromethyl)phenyl)acetamide (C2): white solid, yield 45%, melting point 206–208 °C. 1 H NMR(500MHz,DMSO-d6)δ10.56(s,1H,NH),9.94(s,1H,NH),7.81(s,4H,phenyl H),7.53–7.51(m,2H,phenyl H),7.50–7.47(m,2H,phenyl H),5.84(s,1H,CH),2.01(s,3H,Ar-NHCOCH3).

[0073]

[0074] N-(4-acetamidophenyl)-2-chloro-2-(3-chlorophenyl)acetamide (C3): white solid, yield 41%, melting point 211–213℃. 1 H NMR(500MHz,DMSO-d6)δ10.52(s,1H,NH),9.95(s,1H,NH),7.65–7.64(m,1H,phenyl H),7.54–7.45(m,7H,phenyl H),5.74(d,J=5.0Hz,1H,CH),2.01(s,3H,Ar-NHCOCH3),

[0075]

[0076] N-(4-acetamidophenyl)-2-chloro-2-(3-fluorophenyl)acetamide (C4): white solid, yield 44%, melting point 205–207 °C. 1 H NMR(500MHz,DMSO-d6)δ10.52(s,1H,NH),9.95(s,1H,NH),7.65–7.64(m,1H,phenyl H),7.54–7.45(m,7H,phenyl H),5.74–5.73(d,J=5.0Hz,1H,CH),2.01(s,3H,Ar-NHCOCH3),

[0077] N-(4-acetamidophenyl)-2-chloro-2-(4-bromophenyl)acetamide (C5): white solid, yield 44%. Melting point 213–215 °C. 1 H NMR(500MHz,DMSO-d6)δ9.90(s,1H,NH),9.88(s,1H,NH),7.61–7.58(m,2H,phenyl H),7.50–7.47(m,2H,phenyl H),7.40–7.33(m,2H,phenyl H),7.30–7.26(m,2H,phenyl H),5.09–5.08(d,J=5.0Hz,1H,CH),2.01(s,3H,Ar-NHCOCH3),

[0078]

[0079] N-(4-acetamidophenyl)-2-chloro-2-(4-fluorophenyl)acetamide (C6): white solid, yield 71%, melting point 211–213 °C. 1 H NMR(500MHz,DMSO-d6)δ10.47(s,1H,NH),9.94(s,1H,NH),7.65–7.62(m,2H,phenyl H),7.54–7.47(m,4H,phenyl H),7.28–7.24(m,2H,phenyl H),5.74(s,1H,CH),2.01(s,3H,Ar-NHCOCH3),

[0080]

[0081] N-(4-acetamidophenyl)-2-chloro-2-(4-chlorophenyl)acetamide (C7): white solid, yield 46%, melting point 206–209 °C. 1H NMR(500MHz,DMSO-d6)δ10.51(s,1H,NH),9.94(s,1H,NH),7.62–7.59(m,2H,phenyl H),7.53–7.50(m,2H,phenyl H),7.49–7.47(m,4H,phenyl H)5.75(s,1H,CH),2.01(s,3H,Ar-NHCOCH3).

[0082]

[0083] N-(4-acetamidophenyl)-2-chloro-2-(4-methoxyphenyl)acetamide (C8): white solid, yield 41%, melting point 206–208 °C. 1 H NMR(500MHz,CDCl3)δ9.81(s,1H,NH),9.70(s,1H,NH),7.59–7.53(m,2H,phenyl H),7.53–7.47(m,2H,phenyl H),7.38–7.31(m,2H,phenyl H),6.94–6.88(m,2H,phenyl H),5.58(t,J=5.0Hz,CH),3.79(s,3H,Ar-OCH3),2.08(s,3H,Ar-NHCOCH3).

[0084]

[0085] N-(4-acetamidophenyl)-2-chloro-2-(2-chlorophenyl)acetamide (C9): white solid, yield 45%, melting point 206–208 °C. 1 H NMR(500MHz,CDCl3)δ9.96(s,1H,NH),9.70(s,2H,NH),7.58–7.56(m,2H,phenyl H),7.52–7.47(m,4H,phenyl H),7.35–7.25(m,2H,phenyl H),6.64–6.62(d,J=10.0Hz,1H,OH),5.62–5.61(d,J=5.0Hz,1H,CH),2.01(s,3H,Ar-NHCOCH3).

[0086]

[0087] 2-Chloro-N,2-Diphenylacetamide (C10): White solid, yield 64%, melting point 187–189 °C. 1H NMR(500MHz,DMSO-d6)δ8.44(s,1H,NH),7.58-7.55(m,2H,phenyl H),7.51–7.49(m,2H,phenylH),7.42–7.40(m,3H,phenyl H),7.38–7.33(m,2H,phenyl H),7.19–7.15(m,1H,phenylH),5.50(s,1H,CH).

[0088]

[0089] 2-Chloro-N-phenyl-2-(4-(trifluoromethyl)phenyl)acetamide (C11): white solid, yield 44%, melting point 185–187 °C. 1 H NMR(500MHz,CDCl3)δ8.51(s,1H,NH),7.67–7.62(m,4H,phenyl H),7.57–7.53(m,2H,phenyl H),7.39–7.33(m,2H,phenyl H),7.21–7.17(m,1H,phenyl H),5.53(d,J=5.0Hz,1H,CH).

[0090]

[0091] 2-Chloro-2-(3-chlorophenyl)-N-phenylacetamide (C12): white solid, yield 46%, melting point 185–187 °C. 1 HNMR (500MHz, CDCl3) δ8.70 (s, 1H, NH), 7.51–7.44 (m, 3H, phenyl H), 7.36–7.28 (m, 5H, phenyl H), 7.16–7.13 (t, J = 5.0Hz, 1H, phenyl H), 5.42 (s, 1H, CH).

[0092]

[0093] 2-Chloro-2-(3-fluorophenyl)-N-phenylacetamide (C13): white solid, yield 46%, melting point 186–188 °C. 1 HNMR(500MHz,CDCl3)δ8.51(s,1H,NH),7.66–7.62(m,4H,phenyl H),7.57–7.54(m,2H,phenyl H),7.38–7.34(m,2H,phenyl H),7.21–7.17(m,1H,phenyl H),5.53(s,1H,CH).

[0094]

[0095] 2-Chloro-2-(4-bromophenyl)-N-phenylacetamide (C14): white solid, yield 45%, melting point 188-190℃. 1 HNMR(500MHz,CDCl3)δ8.60(s,1H,NH),7.61–7.57(m,2H,phenyl H),7.39–7.36(m,2H,phenyl H),7.34–7.30(m,2H,phenyl H),7.13–7.09(m,1H,phenyl H),7.91–7.89(m,2H,phenyl H),4.79(d,J=5.0Hz,1H,CH).

[0096]

[0097] 2-Chloro-2-(4-fluorophenyl)-N-phenylacetamide (C15): white solid, yield 65%, melting point 187–189 °C. 1 HNMR(500MHz,DMSO-d6)δ10.38(s,1H,NH),7.63–7.59(m,2H,phenyl H),7.56–7.54(m,2H,phenyl H),7.26–7.22(m,2H,phenyl H),7.11–7.07(m,2H,phenyl H),7.04–7.01(m,1H,phenyl H),5.68(s,1H,CH).

[0098]

[0099] 2-Chloro-2-(4-chlorophenyl)-N-phenylacetamide (C16): white solid, yield 45%, melting point 181–183 °C. 1 HNMR(500MHz,DMSO-d6)δ10.38(s,1H,NH),7.58–7.53(m,4H,phenyl H),7.37–7.53(m,2H,phenyl H),7.26–7.23(m,2H,phenyl H),7.05–7.01(m,1H,phenyl H),5.66(s,1H,CH).

[0100]

[0101] 2-Chloro-2-(4-methoxyphenyl)-N-phenylacetamide (C17): white solid, yield 47%, melting point 181–183 °C. 1H NMR(500MHz,CDCl3)δ9.78(s,1H,NH),7.59–7.56(m,2H,phenyl H),7.42–7.39(m,2H,phenyl H),7.36–7.32(m,2H,phenyl H),7.08–7.05(m,1H,phenyl H), 6.94–6.92 (m, 2H, phenyl H), 5.58–5.57 (t, J = 5.0Hz, 1H, CH), 3.80 (s, 3H, Ar-OCH3).

[0102]

[0103] 2-Chloro-2-(2-chlorophenyl)-N-phenylacetamide (C18): white solid, yield 47%, melting point 183–185 °C. 1 HNMR(500MHz,DMSO-d6)δ10.76(s,1H,NH),7.79–7.76(m,1H,phenyl H),7.63–7.60(m,2H,phenyl H),7.54–7.52(m,1H,phenyl H),7.45–7.43(m,2H,phenyl H),7.36–7.32(m,2H,phenyl H),7.13–7.09(m,1H,phenyl H),6.11(s,1H,CH).

[0104]

[0105] N,2-Di(4-bromophenyl)-2-chlorophenylacetamide (C19): White solid, yield 74%, melting point 196–198 °C. 1 HNMR(500MHz,CDCl3)δ8.41(s,1H,NH),7.55–7.52(m,2H,phenyl H),7.46–7.45(m,2H,phenyl H),7.37–7.35(m,4H,phenyl H),5.44(s,1H,CH).

[0106]

[0107] 2-(4-Bromophenyl)-2-chloro-N-(4-chloro-2-tolyl)acetamide (C20): white solid, yield 47%, melting point 196–198 °C. 1H NMR(500MHz,CDCl3)δ9.29(s,1H,NH),7.73–7.71(d,J=5.0Hz,1H,phenylH),7.57–7.54(m,2H,phenyl H),7.33–7.31(m,2H,phenyl H),7.20–7.16(m,2H,phenylH),5.69(t,J=5.0Hz,1H,CH),2.28(s,3H,Ar-CH3).

[0108]

[0109] 2-(4-bromophenyl)-2-chloro-N-(2,4-difluorophenyl)acetamide (C21): white solid, yield 74%, melting point 197–199 °C. 1 H NMR (500MHz, CDCl3) δ8.59(s,1H,NH),8.26–8.16(m,2H,phenyl H),7.40–7.37(m,2H,phenyl H),7.15–7.11(m,3H,phenyl H),5.47(s,1H,CH).

[0110]

[0111] 2-(4-bromophenyl)-2-chloro-N-(4-fluorophenyl)acetamide (C22): white solid, yield 74%, melting point 192–194 °C. 1 HNMR(500MHz,CDCl3)δ8.70(s,1H,NH),8.27–8.22(m,1H,phenyl H),7.56–7.51(m,2H,phenyl H),7.40–7.38(m,2H,phenyl H),7.16–7.08(m,3H,phenyl H),5.47(s,1H,CH).

[0112]

[0113] 2-(4-bromophenyl)-2-chloro-N-(3-fluorophenyl)acetamide (C23): white solid, yield 44%, melting point 194–196 °C. 1HNMR(500MHz,CDCl3)δ9.68(s,1H,NH),7.61–7.53(m,3H,phenyl H),7.38–7.27(m,4H,phenyl H),6.89–6.81(m,1H,phenyl H),5.50–5.48(d,J=10.0Hz,1H,OH),5.35–5.33(d,J=10.0Hz,1H,CH).

[0114] Synthesis of target compound Z

[0115] (1) Taking the synthesis of target compound Z1 as an example

[0116]

[0117] In a flask containing intermediate E, an appropriate amount of N,N-dimethylformamide solvent was added, followed by 1-acetylpiperazine and potassium carbonate reagent. The solvent was stirred at room temperature for 3 hours, and the reaction was monitored by thin-layer chromatography. The reagent was washed 4–5 times with ammonium chloride solvent, and then dried over anhydrous sodium sulfate. Purification by silica gel column chromatography (dichloromethane / methanol = 5 / 1) yielded the target compound Z1. It was a white solid, 44% yield, with a melting point of 226–228 °C. 1 H NMR(500MHz,DMSO-d6)δ10.39(s,1H,NH),10.08(s,1H,NH),7.56–7.53(m,3H,phenyl H),7.51–7.49(m,3H,phenyl H),7.37–7.33(m,2H,phenyl H),7.31–7.27(m,1H,phenyl H),4.15(s,1H,CH),3.44(s,2H,piperazinyl CONCH2),3.38(s,2H,piperazinyl CONCH2),2.40–2.37(m,2H,piperazinylCHN CH2 ),2.33–2.31(m,2H,piperazinyl CHN CH2 ),2.00(s,3H,Ar-NHCO CH3 ),1.95(s,3H,piperazinyl-NCOCH3); 13C NMR(101MHz,DMSO-d6)δ168.36,167.81,167.75,136.82,134.88,133.63,128.37,1 28.03,127.58,119.56,118.81,73.71,50.71,50.18,23.61,20.88; HRMS(ESI):m / z calcd for C 22 H 26 N4O3[M+H] + 395.2077, found 395.2065.

[0118] The synthesis methods for other target compounds are similar to those for target compound Z1.

[0119] (2) Target compound structure and physicochemical data

[0120]

[0121] Z2, yellow solid, yield 43%, melting point 230–232℃. 1 H NMR (500MHz, DMSO-d6) δ10.58(s,1H,NH),10.06(s,1H,NH),7.75–7.74(d,J=5.0Hz,4H,phenyl H),7.56–7.54(d,J=10.0Hz,2H,phenyl H),7.51–7.49(d,J=10.0Hz,2H,phenyl H),4.34(s,1H,CH),2.88(s,2H,piperazinyl CONCH2),2.72(s,2H,piperazinyl CONCH2),2.43–2.38(m,2H,piperazinylCHN CH2 ),2.34-2.32(t,J=10.0Hz,2H,piperazinyl CHN CH2 ),2.00(s,3H,Ar-NHCO CH3 ),1.96(s,3H,piperazinyl-NCOCH3); 13 C NMR(101MHz,DMSO-d6)δ168.31,168.24,167.98,162.52,142.03,135.48,133.87, 129.60,120.13,119.31,73.44,51.04,50.52,45.71,35.97,30.94,24.04,21.28; 1919F NMR (471 MHz, DMSO-d6) δ 62.67 (s, 3F, CF3); HRMS (ESI): m / z calculated for C 23 H 25 F3N4O3 [M+H] + 463.1951, found 463.1937.

[0122]

[0123] Z3, yellow solid, yield 46%, melting point 223–225 °C. 1 1H NMR (500 MHz, DMSO-d6) δ 10.22 (s, 1H, NH), 9.93 (s, 1H, NH), 7.57–7.56 (m, 1H, phenyl H), 7.52–7.48 (m, 4H, phenyl H), 7.46–7.44 (m, 1H, phenyl H), 7.40–7.39 (m, 2H, phenyl H), 4.08 (s, 1H, CH), 3.46–3.44 (m, 4H, piperazinyl CONCH2CH2), 2.88 (s, 1H, piperazinyl CHN CH ), 2.72 (s, 1H, piperazinylCHN CH ), 2.33–2.31 (d, J = 10.0 Hz, 2H, piperazinyl CHN CH2 ), 2.00 (s, 3H, Ar-NHCO CH3 ), 1.96 (s, 3H, piperazinyl-NCOCH3); 13 13C NMR (126 MHz, DMSO-d6) δ 168.23–168.10 (m), 139.57, 135.47, 133.76, 133.18, 130.45, 128.27 (d, J = 27.5 Hz), 127.58, 120.20, 119.31, 73.73, 51.09, 50.61, 45.65, 40.83, 35.95, 30.91, 24.07, 21.31; HRMS (ESI): m / z calculated forC 22 H 25 ClN4O3 [M+H] + 429.1688, found 429.1674.

[0124]

[0125]

[0126] Z4, yellow solid, yield 45%, melting point 228–230℃. 1 H NMR (500MHz, DMSO-d6) δ10.17(s,1H,NH),9.92(s,1H,NH),7.50–7.49(d,J=5.0Hz,3H,phenyl 7H),7.43–7.39(m,1H,phenylH),7.35–7.31(m,3H,phenyl H),7.18–7.13(m,1H,phenyl H),4.07(s,1H,CH),3.47–3.44(m,4H,piperazinyl CONCH2CH2),2.41–2.38(m,2H,piperazinyl CHN CH2 ),2.34–2.32(m,2H,piperazinyl CHN CH2 ),2.00(s,3H,Ar-NHCO CH3 ),1.96(s,3H,piperazinyl-NCO CH3 ); 13 CNMR(101MHz,DMSO-d6)δ168.29,168.20,168.19,163.47,161.05,139.94,139.87,135.46,133.78,130.53,130.45 ,125.00,120.26,119.35,115.39,115.17,114.96,73.87,51.11,50.63,45.68,40.86,24.07,21.32; HRMS(ESI):m / z calcd for C 22 H 25 FN4O3 413.1983, found 413.1973.

[0127]

[0128] Z5, yellow solid, yield 51%, melting point 224–226℃. 11H NMR (500 MHz, DMSO-d6) δ 10.25 (s, 1H, NH), 9.96 (s, 1H, NH), 7.58–7.56 (m, 2H, phenyl H), 7.50–7.49 (m, 4H, phenyl H), 7.46–7.45 (m, 2H, phenyl H), 4.07 (s, 1H, CH), 2.88 (s, 1H, piperazinyl CONCH), 2.73 (s, 1H, piperazinyl CONCH), 2.54 (s, 6H, piperazinyl C3H6), 2.00 (s, 3H, Ar-NHCOCH3), 1.96 (s, 3H, piperazinyl-NCOCH3); 13 13C NMR (101 MHz, DMSO-d6) δ 168.61, 168.52 (d, J = 9.1 Hz), 162.82, 136.80, 135.71, 134.13, 131.76, 131.27, 121.63, 120.49, 119.64, 73.83, 51.35, 50.79, 45.99, 36.27, 31.24, 24.35, 23.45, 21.59; HRMS (ESI): m / z calcd for C 22 H 25 BrN4O3 [M + H] + 473.1182, found 473.1169.

[0129]

[0130] Z6, yellow solid, yield 48%, melting point 228–230 °C. 1 1H NMR (500 MHz, DMSO-d6) δ 10.12 (s, 1H, NH), 9.90 (s, 1H, NH), 7.54–7.49 (m, 6H, phenyl H), 7.22–7.18 (m, 2H, phenyl H), 5.76 (s, 1H, CH), 4.04 (s, 2H, piperazinyl CONCH2), 3.82 (s, 2H, piperazinyl CONCH2), 2.39–2.35 (m, 2H, piperazinyl CHN CH2 ), 2.32–2.30 (m, 2H, piperazinyl CHN CH2 ), 2.0 (s, 3H, Ar-NHCO CH3 ), 1.96 (s, 3H, piperazinyl-NCOCH3); 1313C NMR (101 MHz, DMSO-d6) δ 168.57, 168.22, 168.13, 163.09, 160.66, 135.29, 133.81, 133.16, 133.13, 130.71, 130.62, 120.09, 119.27, 115.36, 115.15, 73.38, 50.99, 50.48, 45.61, 23.97, 21.21. 19 19F NMR (376 MHz, DMSO-d6) δ -114.30–-114.38 (m, 1F, CF); HRMS (ESI): m / z calcd for C 22 H 25 FN4O3 [M+H] + 413.1983, found 413.1969.

[0131]

[0132] Z7, yellow solid, yield 48%, melting point 221–223 °C. 1 1H NMR (500 MHz, DMSO-d6) δ 10.54 (s, 1H, NH), 10.10 (s, 1H, NH), 7.56–7.54 (m, 4H, phenyl H), 7.51–7.50 (m, 2H, phenyl H), 7.43–7.41 (m, 2H, phenyl H), 4.24 (s, 1H, CH), 3.44 (s, 2H, piperazinyl CONCH2), 3.38 (s, 2H, piperazinyl CONCH2), 2.41–2.36 (m, 2H, piperazinyl CHN CH2) , 2.32–2.28 (m, 2H, piperazinyl CHN CH2 ), 2.01 (s, 3H, Ar-NHCO CH3 ), 1.95 (s, 3H, piperazinyl-NCOCH3); 13 13C NMR (101 MHz, DMSO-d6) δ 168.37, 168.24, 168.20, 136.35, 135.40, 134.00, 132.60, 130.62, 128.46, 120.01, 119.24, 73.06, 55.12, 51.03, 50.50, 45.69, 24.05, 21.31; HRMS (ESI): m / z calcd for C 22 H 25 ClN4O3 [M+H]+ 429.1687, found 429.1674.

[0133]

[0134] Z8, yellow solid, yield 43%, melting point 224–226℃. 1 H NMR(500MHz,DMSO-d6)δ10.03(s,1H,NH),9.89(s,1H,NH),7.52–7.50(m,2H,phenyl H),7.48–7.47(m,2H,phenyl H),7.41–7.40(d,J=5.0Hz,2H,phenyl H),6.93–6.91(d,J=10.0Hz,2H,phenyl H),3.93(s,1H,CH),3.73(s,3H,Ar-COCH3),3.43(s,2H,piperazinyl CONCH2),2.54(s,2H,piperazinylCONCH2),2.38–2.35(m,2H,piperazinyl CHN CH2 ),2.31–2.29(m,2H,piperazinyl CHN CH2 ),2.00(s,3H,Ar-NHCO CH3 ),1.96(s,3H,piperazinyl-NCOCH3); 13 C NMR (126MHz, DMSO-d6) δ169.02, 168.09 (d, J = 10.9Hz), 159.01, 135.16, 133.91, 129.85, 128. 76,120.01,119.21,113.81,73.81,55.10,51.06,50.55,45.59,23.96,21.21; HRMS(ESI):m / z calcd forC 23 H 28 N4O4[M+H] + 425.2183, found 425.2167.

[0135]

[0136] Z9, yellow solid, yield 43%, melting point 223–225℃. 11H NMR (500 MHz, DMSO-d6) δ 10.18 (s, 1H, NH), 9.91 (s, 1H, NH), 7.79–7.77 (m, 1H, phenyl H), 7.54–7.51 (m, 2H, phenyl H), 7.50–7.47 (m, 2H, phenyl H), 7.46–7.45 (m, 1H, phenyl H), 7.40–7.32 (m, 2H, phenyl H), 5.76 (s, 1H, CH), 3.47–3.40 (m, 6H, piperazinyl CONC3H6), 2.44–2.36 (m, 2H, piperazinyl CHN CH2 ), 2.01 (s, 3H, Ar-NHCO CH3 ), 1.96 (s, 3H, piperazinyl-NCOCH3); 13 13C NMR (126 MHz, DMSO-d6) δ 169.87–167.65 (m), 135.58, 134.83, 133.55, 132.16, 131.41–129.20 (m), 127.24, 120.62 (d, J = 49.9 Hz), 70.24, 51.01 (d, J = 57.8 Hz), 24.55, 21.41; HRMS (ESI): m / z calcd for C 22 H 26 ClN4O3 [M + H] + 429.1687, found 429.1673.

[0137]

[0138] Z10, yellow solid, yield 49%, melting point 211–213 °C. 1 1H NMR (500 MHz, DMSO-d6) δ 10.23 (s, 1H, NH), 9.99 (s, 1H, NH), 7.54–7.51 (m, 3H, phenyl H), 7.50–7.47 (m, 3H, phenyl H), 7.36–7.33 (m, 2H, phenyl H), 7.31–7.27 (m, 1H, phenyl H), 4.04 (s, 1H, CH), 2.88 (s, 2H, piperazinylCONCH2), 2.72 (s, 2H, piperazinyl CONCH2), 2.54–2.52 (m, 2H, piperazinyl CHN CH2 ), 2.47–2.44 (m, 2H, piperazinyl CHN CH2 ),2.29(s,3H,Ar-NHCO CH3 ),2.00(s,3H,piperazinyl-NCH3); 13 CNMR(126MHz,DMSO-d6)δ168.61(d,J=25.5Hz),168.41,136.57,135.43,134.01,132.74,130.63,128.58,11 9.80(d,J=90.8Hz),73.84,57.73,54.43,50.34,45.32,23.71(d,J=103.1Hz),19.44,13.77; HRMS(ESI):m / z calcd for C 21 H 26 N4O2[M+H] + 367.2128, found 367.2116.

[0139]

[0140] Z11, white solid, yield 49%, melting point 216–218℃. 1 H NMR(400MHz,DMSO-d6)δ10.26(s,1H,NH),9.97(s,1H,NH),7.71(s,4H,phenyl H),7.47–7.44(m,4H,phenyl H),4.06(s,1H,CH),2.34–2.12(m,8H,piperazinyl C4H8),2.12(s,3H,Ar-NHCO CH3 ),1.99(s,3H,piperazinyl-NCH3); 13 C NMR (126MHz, DMSO-d6) δ168.09(d,J=5.0Hz),142.29,135.39,133.75(d,J=45.36Hz),129.45,125.30,125.27,120.10,119.32 74.31,54.65,50.73,45.75,23.98; 19 FNMR(376MHz,DMSO-d6)δ-60.95(s,3F,CF3); HRMS(ESI):m / z calcd for C 22 H 25 F3N4O2[M+H] + 435.1988, found 435.2002.

[0141]

[0142] Z12, yellow solid, yield 46%, melting point 207–209℃. 1 H NMR(500MHz,DMSO-d6)δ10.19(s,1H,NH),9.96(s,1H,NH),7.52–7.51(m,3H,phenyl H),7.50–7.49(m,3H,phenyl H),7.42–7.40(m,2H,phenyl H),3.99(s,1H,CH),2.41–2.17(m,8H,piperazinyl C4H8),2.13(s,3H,Ar-NHCO CH3 )2.00(s,3H,piperazinyl-NCH3); 13 C NMR(126MHz,DMSO-d6)δ168.41(d,J=44.2Hz),136.64,135.38,133.94,132.62,13 0.54,128.50,120.10,119.35,74.14,54.70,50.80,45.81,24.08; HRMS(ESI):m / z calcd for C 21 H 26 ClN4O2[M+H] + 401.1738, found 401.1728.

[0143]

[0144] Z13, yellow solid, yield 48%, melting point 215–218℃. 1 H NMR(500MHz,DMSO-d6)δ10.48(s,1H,NH),10.12(s,1H,NH),7.57–7.54(m,2H,phenyl H),7.52–7.50(m,2H,phenyl H),7.38–7.34(m,3H,phenyl H),7.13–7.09(m,1H,phenyl H),4.15(s,1H,CH),2.44–2.29(m,8H,piperazinyl C4H8),2.15(s,3H,Ar-NHCO CH3 ),2.01(s,3H,piperazinyl-NCH3); 1313C NMR (126 MHz, DMSO-d6) δ 168.38 (d, J = 21.2 Hz), 140.69 (d, J = 7.2 Hz), 135.42, 134.06, 130.36 (d, J = 8.1 Hz), 124.93, 119.68 (d, J = 89.5 Hz), 73.99, 54.61, 50.63, 45.65, 24.07; HRMS (ESI): m / z calcd for C 21 H 25 O2N4F [M+H] + 385.2034, found 385.2020.

[0145]

[0146]

[0147] Z14, yellow solid, yield 43%, melting point 214–216 °C. 1 1H NMR (500 MHz, DMSO-d6) δ 10.12 (s, 1H, NH), 9.93 (s, 1H, NH), 7.52–7.50 (m, 3H, phenyl H), 7.49–7.47 (m, 3H, phenyl H), 7.34–7.33 (m, 1H, phenyl H), 7.30–7.27 (m, 1H, phenyl H), 3.97 (s, 1H, CH), 2.88 (s, 2H, piperazinyl CONCH2), 2.72 (s, 2H, piperazinyl CONCH2), 2.38 (s, 2H, piperazinyl CHN CH2 ), 2.19 (s, 2H, piperazinyl CHN CH2 ), 2.00 (s, 3H, Ar-NHCO CH3 ), 1.90 (s, 3H, piperazinyl-NCH3); 13 13C NMR (101 MHz, DMSO-d6) δ 169.08, 168.28, 137.66, 135.31, 134.13, 128.44, 127.97, 120.03, 119.37, 75.00, 56.23, 54.71, 50.88, 45.81, 24.06, 18.73; HRMS (ESI): m / z calcd for C 21 H 25 N4O2Br [M+H] + 445.1233, found 445.1216.

[0148]

[0149] Z15, yellow solid, yield 43%, melting point 216–218℃. 1 H NMR(500MHz,DMSO-d6)δ10.35(s,1H,NH),10.09(s,1H,NH),7.56–7.49(m,6H,phenyl H),7.17–7.13(m,2H,phenyl H),4.05(s,1H,CH),2.33(s,8H,piperazinyl C4H8),2.11(s,3H,Ar-NHCO CH3 ),2.01(s,1H,piperazinyl-NCH3); 13 C NMR(101MHz,DMSO-d6)δ168.95,168.34,163.10,160.67,135.30,134.06,133.83,133.8 0,130.68,130.60,120.00,119.36,115.31,115.10,73.88,54.66,50.74,45.75,24.02; 19 FNMR(376MHz,DMSO-d6)δ-114.49–-114.57(m,1F,CF).HRMS(ESI):m / z calcd for C 21 H 25 FN4O2[M+H] + 385.2034, found 385.2022.

[0150]

[0151] Z16, yellow solid, yield 53%, melting point 212–214℃. 1 H NMR(500MHz,DMSO-d6)δ10.21(s,1H,NH),10.08(s,1H,NH),7.54–7.48(m,4H,phenyl H),7.42–7.41(d,J=5.0Hz,2H,phenylH),6.90–6.88(m,2H,phenyl H),3.94(s,1H,CH),3.74–3.68(m,4H,piperazinyl COCH2),2.41–2.31(m,4H,piperazinyl CHN CH2) ,2.17(s,3H,Ar-NHCO CH3), 2.01 (s, 1H, piperazinyl - NCH3); 13 C NMR (126 MHz, DMSO - d6) δ 169.24, 168.09, 158.91, 135.13, 134.08, 129.59 (d, J = 45.0 Hz), 119.81, 119.16, 113.71, 74.05, 54.80 (d, J = 75.0 Hz), 50.56, 45.51, 23.95; HRMS (ESI): m / z calcd for C 21 H 25 ClN4O2 [M + H] + 400.1738, found 401.2805.

[0152]

[0153] Z17, yellow solid, yield 47%, melting point 213–215 °C. 1 H NMR (500 MHz, DMSO - d6) δ 9.44 (s, 1H, NH), 7.74 (s, 1H, NH), 7.51–7.48 (m, 2H, phenyl H), 7.46–7.44 (m, 2H, phenyl H), 7.42–7.40 (m, 1H, phenyl H), 7.33–7.31 (m, 1H, phenyl H), 7.24–7.23 (m, 2H, phenyl H), 4.84 (s, 1H, CH), 2.62–2.50 (m, 8H, piperazinyl C4H8), 2.29 (s, 3H, Ar - NHCO CH3 ), 2.08 (s, 3H, piperazinyl - NCH3); 13 C NMR (126 MHz, DMSO - d6) δ 169.07, 168.70, 135.78, 134.75, 133.69, 132.56, 130.73, 130.21, 129.63, 127.00, 120.85, 120.35, 70.11, 55.62, 45.96, 24.47; HRMS (ESI): m / z calcd for C 21 H 25 ClN4O2 [M + H] + 401.1738, found 401.1734.

[0154]

[0155] Z18, white solid, yield 72%, melting point 212–214℃. 1 H NMR (500MHz, CDCl3) δ9.01(s,1H,NH),7.64–7.62(d,J=10.0Hz,2H,phenyl H),7.56–7.54(d,J=10.0Hz,2H,phenyl H),7.49–7.47(d,J=10.0Hz,2H,phenyl H),7.36–7.32(t,J=10.0Hz,3H,phenyl H),7.16–7.12(m,1H,phenyl H),4.13(s,1H,CH),3.69–3.68(d,J=5.0Hz,2H,piperazinylCONCH2),3.55–3.53(t,J=10.0Hz,2H,piperazinyl CONCH2),2.53–2.48(m,4H,piperazinyl CHN CH2CH2 ),2.06(s,3H,piperazinyl-NCOCH3); 13 C NMR (101MHz, CDCl3) δ169.11,167.96,137.26,129.56,129.30,129.24,129.01,1 25.88,124.88,119.69,75.53,51.48,51.22,46.33,41.48,21.38; HRMS(ESI):m / z calcd for C 20 H 23 N3O2[M+H] + 338.1863, found 338.1849.

[0156]

[0157] Z19, white solid, yield 65%, melting point 216–218℃. 1 H NMR(500MHz,CDCl3)δ9.03(s,1H,NH),7.57–7.55(d,J=10.0Hz,2H,phenyl H),7.36–7.33(m,6H,phenyl H),7.14–7.10(t,J=10.0Hz,1H,phenyl H),4.00(s,1H,CH),3.66–3.63(m,2H,piperazinyl CONCH2),3.53–3.50(t,J=10.0Hz,2H,piperazinyl CONCH2),2.50–2.45(m,4H,piperazinyl CHN CH2CH2 ), 2.06 (s, 3H, piperazinyl-NCOCH3); 13 C NMR (101 MHz, CDCl3) δ 169.09, 168.93, 137.51, 134.62, 129.20, 128.97, 128.79, 124.58, 119.64, 76.16, 51.59, 51.30, 46.39, 41.54, 21.39; 19 F NMR (376 MHz, CDCl3) δ –62.67 (s, 3F, CF3); HRMS (ESI): m / z calcd for C 21 H 22 F3N3O2 [M+H] + 406.1736, found 406.1722.

[0158]

[0159] Z20, yellow solid, yield: 50%, melting point 211–213 °C. 1 H NMR (500 MHz, CDCl3) δ 9.02 (s, 1H, NH), 7.56–7.54 (d, J = 10.0 Hz, 2H, phenyl H), 7.35–7.30 (m, 4H, phenyl H), 7.29–7.27 (m, 1H, phenyl H), 7.24–7.21 (m, 1H, phenyl H), 4.00 (s, 1H, CH), 3.69–3.65 (m, 2H, piperazinyl CON CH2 ), 3.53–3.50 (m, 2H, piperazinyl CON CH2 ), 2.50–2.45 (m, 4H, piperazinyl CHN CH2CH2 ), 2.06 (s, 3H, piperazinyl-NCOCH3); 13 C NMR (101 MHz, CDCl3) δ 169.11, 168.18, 137.33, 136.43, 134.84, 132.11, 130.71, 130.20, 119.26, 119.10, 119.02, 127.33, 124.79, 119.72, 77.48, 75.49, 51.49, 51.23, 46.35, 41.50, 21.39; HRMS (ESI): m / z calcd for C 20 H 22 ClN3O2 [M+H] +372.1473, found 372.1460.

[0160]

[0161] Z21, yellow solid, 70% yield, melting point 214–216℃. 1 H NMR(500MHz,CDCl3)δ8.98(s,1H,NH),7.56–7.54(d,J=10.0Hz,2H,phenyl H),7.36–7.31(m,3H,phenyl H),7.15–7.11(m,2H,phenyl H),7.09–7.01(m,2H,phenyl H),4.02(s,1H,CH),3.70–3.66(m,2H,piperazinyl CONCH2),3.54–3.52(t,J=5.0Hz,2H,piperazinyl CONCH2),2.52–2.46(m,4H,piperazinyl CHN CH2CH2 ),2.07(s,3H,piperazinyl-NCOCH3); 13 C NMR (101MHz, CDCl3) δ169.12,168.26,164.18,161.73,137.34,136.90,130.58,130.49,129.26,124.90,1 24.87,124.78,119.69,116.00,115.97,115.78,115.76,75.59,75.58,51.53,51.26,46.36,41.51,21.39; 19 F NMR (376MHz, CDCl3) δ-111.84–-111.91 (m, 1F, CF); HRMS (ESI): m / z calcdfor C 20 H 22 N3O2F[M+H] + 356.1768, found 356.1763.

[0162]

[0163] Z22, yellow solid, yield 72%, melting point 212–214℃. 11H NMR (500 MHz, CDCl3) δ 8.97 (s, 1H, NH), 7.55–7.54 (d, J = 5.0 Hz, 2H, phenyl H), 7.51–7.49 (m, 2H, phenyl H), 7.35–7.32 (t, J = 10.0 Hz, 3H, phenyl H), 7.23–7.21 (m, 2H, phenyl H), 7.15–7.12 (t, J = 10.0 Hz, 1H, phenyl H), 4.01 (s, 1H, CH), 3.69–3.66 (m, 2H, piperazinyl CONCH2), 3.53–3.51 (t, J = 5.0 Hz, 2H, piperazinyl CONCH2), 2.50–2.46 (m, 4H, piperazinyl CHN CH2CH2 ), 2.06 (s, 3H, piperazinyl-NCOCH3); 13 13C NMR (126 MHz, CDCl3) δ 169.11, 168.31, 137.36, 133.46, 132.15, 130.73, 129.29, 124.79, 122.95, 119.68, 75.43, 51.50, 51.24, 6.34, 41.53, 21.39; HRMS (ESI): m / z calcd for C 20 H 22 BrN3O2 [M + H] + 416.0968, found 416.0954.

[0164]

[0165] Z23, yellow solid, yield 50%, melting point 216–218 °C. 1 1H NMR (500 MHz, CDCl3) δ 9.02 (s, 1H, NH), 7.56–7.54 (m, 2H, phenyl H), 7.35–7.30 (m, 4H, phenyl H), 7.15–7.11 (m, 1H, phenylH), 7.07–7.03 (m, 2H, phenyl H), 4.02 (s, 1H, CH), 3.68–3.66 (m, 2H, piperazinyl CONCH2), 3.53–3.51 (t, J = 5.0 Hz, 2H, piperazinyl CONCH2), 2.49–2.45 (m, 4H, piperazinylCHN CH2CH2 ), 2.06 (s, 3H, piperazinyl-NCOCH3);13 13C NMR (101 MHz, CDCl3) δ 169.08, 168.60, 161.62, 137.34, 130.76, 130.68, 129.24, 124.70, 119.60, 116.05, 115.84, 77.42, 75.22, 51.48, 51.19, 46.34, 41.49, 21.35; 19 19F NMR (376 MHz, CDCl3) δ -112.79–-112.86 (m, 1F, CF); HRMS (ESI): m / z: calcd for C 20 H 22 FN3O2 [M+H] + 356.1768, found 356.1760.

[0166]

[0167] Z24, yellow solid, yield 71%, melting point 212–214 °C. 1 1H NMR (500 MHz, CDCl3) δ 8.98 (s, 1H, NH), 7.56–7.54 (m, 2H, phenyl H), 7.36–7.32 (m, 4H, phenyl H), 7.30–7.27 (m, 2H, phenyl H), 7.15–7.11 (t, J = 5.0 Hz, 1H, phenyl H), 4.02 (s, 1H, CH), 3.70–3.66 (m, 2H, piperazinyl CONCH2), 3.54–3.51 (t, J = 5.0 Hz, 2H, piperazinyl CONCH2), 2.50–2.44 (m, 4H, piperazinyl CHN CH2CH2 ), 2.06 (s, 3H, piperazinyl-NCOCH3); 13 13C NMR (101 MHz, CDCl3) δ 169.10, 168.39, 137.35, 134.76, 132.93, 130.41, 129.28, 129.18, 124.77, 119.66, 75.34, 51.49, 51.22, 46.37, 41.52, 21.40; HRMS (ESI): m / z calcd for C 20 H 22 ClN3O2 [M+H]+ 372.1473, found 372.1459.

[0168]

[0169] Z25, white solid, yield 68%, melting point 214–216℃. 1 H NMR (400MHz, CDCl3) δ9.04(s,1H),7.60–7.56(m,2H,phenyl H),7.37–7.33(m,2H,phenyl H),7.29–7.26(m,2H,phenyl H),7.16–7.11(m,1H,phenyl H),6.91–6.88(m,2H,phenyl H),3.97(s,1H,CH),3.80(s,3H,Ar-OCH3),3.69–3.66(m,2H,piperazinyl CONCH2),3.53(t,J=4.0Hz,2H,piperazinylCONCH2),2.51–2.46(m,4H,piperazinyl CHN CH2CH2 ),2.08(s,3H,piperazinyl-NCOCH3); 13 CNMR (101MHz, CDCl3) δ169.50,168.27,159.15,138.87,129.99,128.83,123.71, 119.68,113.93,73.91,55.21,51.15,50.64,45.71,40.90,21.32HRMS(ESI):m / z calcd forC 21 H 25 N3O3[M+H] + 368.1968, found 368.1961.

[0170]

[0171] Z26, yellow solid, 70% yield, melting point 213–215℃. 11H NMR (500 MHz, CDCl3) δ 9.21 (s, 1H, NH), 7.58–7.55 (m, 2H, phenyl H), 7.42–7.37 (m, 2H, phenyl H), 7.34–7.30 (m, 3H, phenyl H), 7.26–7.24 (m, 2H, phenyl H), 7.13–7.10 (m, 1H, phenyl H), 4.82 (s, 1H, CH), 3.71–3.59 (m, 2H, piperazinyl CONCH2), 3.51–3.49 (t, J = 5.0 Hz, 2H, piperazinyl CONCH2), 2.58–2.50 (m, 4H, piperazinyl CHN CH2CH2 ), 2.04 (s, 3H, piperazinyl-NCOCH3); 13 13C NMR (126 MHz, CDCl3) δ 169.04, 168.28, 137.45, 135.52, 132.20, 130.64, 130.03 (d, J = 52.5 Hz), 129.21, 127.17, 124.66, 119.59, 70.25, 51.17, 50.72, 46.58, 41.72, 21.35; HRMS (ESI): m / z calcd for C 20 H 22 ClN3O2 [M + H] + 372.1473, found 372.1469.

[0172]

[0173] Z27, white solid, yield 67%, melting point 216–218 °C. 1 1H NMR (500 MHz, CDCl3) δ 9.02 (s, 1H, NH), 7.50–7.49 (m, 2H, phenyl H), 7.44–7.43 (d, J = 5.0 Hz, 4H, phenyl H), 7.20–7.19 (m, 2H, phenyl H), 4.01 (s, 1H, CH), 3.68–3.63 (m, 2H, piperazinyl CONCH2), 3.53–3.51 (t, J = 5.0 Hz, 2H, piperazinyl CONCH2), 2.48–2.44 (m, 4H, piperazinyl CHN CH2CH2 ]), 2.06 (s, 3H, piperazinyl-NCOCH3); 13C NMR (126MHz, CDCl3) δ169.10, 168.39, 136.44, 133.10, 132.25 (d, J = 4.3Hz), 132.21, 130 .75,123.09,121.22,117.36,75.35,51.48,51.23,46.35,41.50,21.40; HRMS(ESI):m / z calcd for C 20 H 21 Br2N3O2[M+H] + 493.2219, found 494.0479.

[0174]

[0175] Z28, white solid, yield 65%, melting point 220–222℃. 1 H NMR(500MHz,CDCl3)δ8.95(s,1H,NH),7.52–7.49(m,3H,Phenyl H),7.22–7.20(m,2H,Phenyl H),7.05–7.01(m,2H,PhenylH),4.01(s,1H,CH),3.71–3.63(m,2H,piperazinyl CON CH2 ),3.55–3.49(m,2H,piperazinyl CON CH2 ),2.53–2.43(m,4H,piperazinyl CHN CH2CH2 ),2.06(s,3H,piperazinyl-NCOCH3),1.25(s,3H,Ar CH3 ); 13 C NMR (126MHz, CDCl3) δ169.12, 168.29, 160.54, 158.60, 133.28 (d, J = 17.3Hz), 132.18, 130.72, 123 .02,121.38(d,J=7.9Hz),115.96(d,J=22.5Hz),75.27,51.47,51.23,46.33,41.46,29.84,21.46.

[0176]

[0177] Z29, white solid, yield 64%, melting point 226–228℃. 11H NMR (500 MHz, CDCl3) δ 9.47 (s, 1H, NH), 8.26–8.23 (m, 1H, phenyl H), 7.51–7.48 (m, 2H, phenyl H), 7.21–7.19 (m, 2H, phenyl H), 7.13–7.09 (m, 2H, phenyl H), 4.08 (s, 1H, CH), 3.72–3.62 (m, 2H, piperazinyl CONCH2), 3.57–3.49 (m, 2H, piperazinyl CONCH2), 2.52–2.42 (m, 4H, piperazinyl CHN CH2CH2 ), 2.05 (s, 3H, piperazinyl-NCOCH3); 13 13C NMR (126 MHz, CDCl3) δ 169.02, 168.38, 153.60, 151.67, 132.90, 132.03, 130.79, 125.75 (d, J = 10.1 Hz), 124.73 (d, J = 8.0 Hz), 122.89, 121.47, 75.11, 51.26, 51.01, 46.27, 41.42, 21.28.

[0178]

[0179] Z30, white solid, yield 71%, melting point 216–218 °C. 1 1H NMR (500 MHz, CDCl3) δ 8.26–8.23 (m, 1H, phenyl H), 7.52–7.49 (m, 4H, phenyl H), 7.22–7.20 (m, 2H, phenyl H), 7.05–7.01 (m, 2H, phenyl H), 4.01 (s, 1H, CH), 3.69–3.65 (m, 2H, piperazinyl CONCH2), 3.54–3.51 (m, 2H, piperazinyl CONCH2), 2.48–2.43 (m, 4H, piperazinyl CHN CH2CH2 ), 2.06 (s, 3H, piperazinyl-NCOCH3); 1313C NMR (126 MHz, CDCl3) δ 173.59, 169.10, 168.29, 160.52, 158.58, 133.24, 132.16, 130.71, 122.99, 121.37 (d, J = 7.9 Hz), 115.94 (d, J = 22.6 Hz), 75.26, 55.55, 46.32, 41.45, 21.45; HRMS (ESI): m / z calcd for C 20 H 21 BrFN3O2 [M+H] + 433.0869, found 434.1728.

[0180]

[0181] Z31, white solid, yield 54%, melting point 216–218 °C. 1 1H NMR (500 MHz, CDCl3) δ 9.07 (s, 1H, NH), 7.50–7.48 (m, 3H, phenyl H), 7.29–7.24 (m, 1H, phenyl H), 7.21–7.16 (m, 3H, phenylH), 6.84–6.80 (m, 1H, Phenyl H), 4.01 (s, 1H, CH), 3.70–3.62 (m, 2H, piperazinyl CONCH2), 3.52–3.50 (t, J = 5.0 Hz, 2H, piperazinyl CONCH2), 2.51–2.41 (m, 4H, piperazinylCHN CH2CH2 ), 2.05 (s, 3H, piperazinyl-NCOCH3); 13 13C NMR (126 MHz, CDCl3) δ 169.05, 168.39, 164.09, 162.14, 138.80 (d, J = 10.8 Hz), 133.00, 132.17, 130.94–130.07 (m), 123.06 114.82 (d, J = 2.7 Hz), 114.47 (d, J = 21.3 Hz), 107.18 (d, J = 26.3 Hz), 75.29, 46.29, 41.44, 21.35; 19 19F NMR (376 MHz, CDCl3) δ -110.83– -110.88 (m, 1F, CF); HRMS (ESI): m / z calcd forC 20 H 21 BrFN3O2 [M+H] +433.0869, found 434.1728.

[0182] Bioactivity assay method for target compounds

[0183] Fungal antibacterial activity test

[0184] The mycelial growth rate method was used to test the activity of the target compound against Monilinia fructicola at a concentration of 50 mg / L for preliminary screening.

[0185] EC 50 Determination of value

[0186] Based on the preliminary screening data, the fungal inhibition rates of the target compounds with good activity were further determined at concentrations of 50, 25, 12.5, 6.25, and 3.125 mg / L. The diameter of the hyphae was measured using the cross-multiplication method, and the inhibition rate was calculated using the same method as above. Finally, the virulence regression equation y = a + bx and EC were calculated using SPSS 26.0 software. 50 Values ​​(x represents the inverse ratio of concentration, y represents the statistical probability of inhibition rate at the corresponding concentration) and correlation coefficients (R²) 2 ).

[0187] Inhibition test on spore germination of brown rot fungus in stone fruit

[0188] Under aseptic conditions, the mycelial cake of *Pseudomonas aeruginosa*, the causal agent of brown rot of stone fruits, was inoculated into a petri dish containing PDA medium using a sterile needle. The petri dish was then incubated in an incubator with alternating light and dark conditions for 7 days to induce the production of conidia from the conidial-bearing mycelia. Once the conidia covered the entire petri dish, the conidial-bearing mycelia were transferred into a 10 mL centrifuge tube using a sterile syringe, and 5 mL of sterile water was added to the tube. The centrifuge tube was then shaken for 15 minutes to dislodge a large number of conidia into the water. The mycelia were then filtered through gauze multiple times to obtain a pure spore suspension. The concentration of the spore suspension was then adjusted by centrifugation or dilution to obtain a final concentration of 1 × 10⁻⁶. 5 Spore / mL suspension. On a clean bench, add 3 mL of PDB solution to a clean test tube, and add the prepared target compound at different concentrations and the positive control drug (with 1% DMSO as a blank control). Incubate the centrifuge tubes at 25℃ (180 rpm) for 3–4 h. Observe the germination number of brown rot spores from the control and different concentration treatments under a stereomicroscope, calculate the spore germination rate, and perform three replicates.

[0189] Tests on the prevention and control of brown rot in pear fruit after harvest.

[0190] Take 10 mg of compound Z 31The azoxystrobin compound was added to 1 mL of DMSO to prepare a stock solution, and then different amounts of distilled water were added to obtain solvents with final concentrations of 100 and 200 mg / L, respectively. Pears that were uniform in size, had no blemishes on the peel, had uniform color, and were naturally ripe were selected, washed with sterile distilled water, air-dried, and then sprayed evenly with 75% alcohol solution.

[0191] Protective activity of compounds on fruit

[0192] After drying, the pesticide was evenly sprayed onto the surface of the pears until it dripped. Simultaneously, the blank control group was sprayed with the same dose of sterile distilled water containing 1% DMSO. The sprayed pears were stored in an incubator for 24 hours. After 24 hours, holes with a diameter of 5 mm and a depth of 1–2 mm were formed on the surface of the pears using a sterile punch. A 5 mm diameter piece of drupe rot fungus was then inserted into the hole using a sterile needle. The inoculated pears were placed in a constant temperature incubator at 25℃ and 90% humidity for 5 days. The diameter of the lesions was measured using a cross-hatching method, with each group's experiment repeated three times.

[0193] Control effect (%) = (Control spot diameter - Experimental spot diameter) / Control spot diameter.

[0194] Therapeutic activity of compounds on fruits

[0195] The fungal cake was inoculated onto the fruit surface in a clean bench and incubated at 25°C for 24 hours. Then, the reagent and sterile distilled water containing 1% DMSO were sprayed using the same administration method. Other experimental procedures and the calculation methods for control effects were the same as those for protective effects.

[0196] Antibacterial activity data of the target compound

[0197] Based on the experimental steps described above, the in vitro bioactivity of all target compounds against the brown rot pathogen of stone fruit was tested using the growth rate assay method. The results are shown in Tables 1 and 2.

[0198] Table 1. Antimicrobial activity test results of compounds Z1-Z31 against *Brachys spp.* at a concentration of 50 mg / L. a

[0199]

[0200]

[0201] “a” indicates that each experiment is repeated three times.

[0202] Table 2 shows the EC50 activity of some compounds against *Pseudomonas aeruginosa*, the causal agent of brown rot in stone fruits. 50 a

[0203] serial number <![CDATA[EC 50 (μg / mL)]]> Poisoning equation <![CDATA[R 2 ]]> <![CDATA[Z4]]> 49.4±6.9 y = 1.2946x + 2.8074 0.953 <![CDATA[Z5]]> 25.0±0.6 y = 1.6804x + 2.6499 0.912 <![CDATA[Z9]]> 90.0±13.6 y = 0.9617x + 3.1207 0.949 <![CDATA[Z 10 ]]> 117.2±5.4 y = 1.1687x + 2.5819 0.922 <![CDATA[Z 13 ]]> 53.0±6.6 y = 1.4184x + 2.5542 0.939 <![CDATA[Z 26 ]]> 44.1±2.6 y = 1.406x + 2.6877 0.925 <![CDATA[Z 14 ]]> 50.3±1.2 y = 1.292x + 2.8013 0.938 <![CDATA[Z 18 ]]> 76.5±7.5 y = 1.3371x + 2.4812 0.957 <![CDATA[Z 19 ]]> 42.4±1.0 y = 1.4074x + 2.7102 0.925 <![CDATA[Z 20 ]]> 43.3±2.4 y = 1.4336x + 2.6542 0.979 <![CDATA[Z 21 ]]> 18.0±0.8 y = 1.3328x + 3.3277 0.935 <![CDATA[Z 22 ]]> 12.5±0.6 y = 1.6242x + 3.2167 0.967 <![CDATA[Z 23 ]]> 41.2±2.4 y = 1.9053x + 1.9239 0.967 <![CDATA[Z 28 ]]> 22.0±0.4 y = 1.4026x + 3.1162 0.915 <![CDATA[Z 29 ]]> 16.2±0.6 y = 1.4006x + 3.3078 0.895 <![CDATA[Z 30 ]]> 18.8±2.2 y = 1.3567x + 3.2716 0.992 <![CDATA[Z 31 ]]> 11.8±0.2 y = 1.5422x + 3.3454 0.994

[0204] “a” indicates that each experiment is repeated three times.

[0205] As shown in Table 1, in preliminary in vitro antifungal tests, these compounds, at a concentration of 50 mg / L, exhibited significant inhibitory effects against *Brachys sclerotium*, the causal agent of brown rot in stone fruits. For example, compound Z... 21 Z 22 The activities of these compounds all reached over 90%, so they were selected for further activity screening against *Pseudomonas aeruginosa*, the causal agent of brown rot in stone fruits. Furthermore, some compounds showed inhibition rates of up to 64.3% against other fungal species. When diamide compounds were replaced with monoamide compounds, the activity of the compounds was correspondingly improved, indicating that the optimization of the compound structure was effective. However, when electron-donating groups were attached to the piperazine group, as shown in Table 1, there was no significant improvement in the activity of the compounds; the activity of most compounds was not ideal. According to Z... 22 EC 50 =12.5mg / L, in Z 22 Based on the structure, di-substitution and mono-substitution are carried out to obtain compounds with higher activity. For mono- and di-substituted compounds Z... 27 –Z 31 Preliminary screening for resistance to brown rot pathogens of stone fruit is shown in Table 1. At a concentration of 50 mg / L, Z... 27 –Z 31 The inhibitory activity ranged from 62.1% to 91.0%, further demonstrating the effectiveness of compound structure optimization. While some compounds exhibited good inhibition rates at 50 mg / L, the EC50 was lower due to the lower concentrations. 50 The value is high, and the inhibition rate is significantly reduced. As shown in Table 2, Z 31 EC50 against brown rot fungus of stone fruit 50 The value was 11.8 mg / L. 21 Z 22 Z 28 and Z 29 EC50 against brown rot fungus of stone fruit 50 The values ​​were 18.0, 12.5, 22.0 and 16.2 mg / L, respectively.

[0206] Table 3. Compound Z at different concentrations 31 Information related to the treatment of brown rot spores of stone fruit a

[0207] CK <![CDATA[1×EC 50 ]]> <![CDATA[2×EC 50 ]]> <![CDATA[4×EC 50 ]]> Number of germinating spores 328 279 155 68 Total number of spores 379 412 370 373 Spore germination rate 86.49％ 67.66％ 41.87％ 18.11％ Inhibition rate -- 21.69％ 51.54％ 79.06％

[0208] a The value is the average of three repetitions; "--" indicates no data.

[0209] Table 4. Information on spores of brown rot fungus in stone fruits treated with different concentrations of the commercial pesticide azoxystrobin. a

[0210] CK <![CDATA[1×EC 50 ]]> <![CDATA[2×EC 50 ]]> <![CDATA[4×EC 50 ]]> Number of germinating spores 328 216 81 57 Total number of spores 379 434 302 424 Spore germination rate 86.49％ 49.79％ 26.85％ 13.48％ Inhibition rate -- 43.32％ 68.92％ 84.40％

[0211] a The value is the average of three repetitions; "--" indicates no data.

[0212] As shown in Table 3, different concentrations (1×EC) 50 2×EC 50 and 4×EC 50 )Z 31 The germination rate of spores treated for 4 hours was significantly lower than that of the blank control group (86.5%). The germination rates of spores at different concentrations were 67.6%, 41.8%, and 18.1%, respectively, which were higher than those of the positive control (49.7%, 26.9%, and 13.5%) (Table 4). These results confirm that Z... 31 It has good in vitro antifungal effects.

[0213] Table 5 Compound Z 31 The protective and curative effects of azoxystrobin on brown rot of pear fruit after harvest. f

[0214]

[0215] f Values ​​are the mean ± standard deviation of three replicates; "--" indicates no data. abcde IBM SPSS Statistics 20 data processing software was used, and Duncan's test was performed to analyze statistical significance. Different letters in the same column indicate statistically significant differences (p<0.05).

[0216] As shown in Table 5, the incidence and diameter of brown rot in the untreated control fruit were significantly higher than those in the Z control. 31 Processed pears. (Z) 31 The protective activities at concentrations of 100 and 200 mg / L were 60.6% and 79.4%, respectively, while the therapeutic activities at concentrations of 100 and 200 mg / L were 55.1% and 70.5%, respectively. At the same concentration, Z... 31 The therapeutic and protective activities against pears were slightly lower than those of azoxystrobin; however, spore germination rate data showed that Z... 31 It exhibited better inhibition rates than azoxystrobin at both low and high concentrations. Systematic analysis revealed compound Z. 31 These active compounds exhibit strong comprehensive activity in both in vitro and in vivo tests, and show potential application value, particularly in the prevention and control of diseases in post-harvest fruits.

Claims

1. The application of a class of phenylacetamide derivatives containing α-piperazine groups in the control of brown rot in stone fruits, wherein the derivatives have the following structural formula: R1 is hydrogen, trifluoromethyl, methoxy or halogen, R2 is hydrogen, acetamido, methyl or halogen, and R3 is methyl or acetyl.

2. Application of a class of phenylacetamide derivatives containing α-piperazine groups in the prevention and control of brown rot in stone fruits after harvest, wherein the derivatives have the following structural formula: R1 is hydrogen, trifluoromethyl, methoxy or halogen, R2 is hydrogen, acetamido, methyl or halogen, and R3 is methyl or acetyl.

3. Use according to claim 1 or 2, characterized in that: The substituent position of R1 is single substitution at ortho, meta or para position, and the substituent of R2 is hydrogen, acetamido, methyl or halogen.

4. Use according to claim 3, characterized in that: The R2 substituted phenyl is single substitution at ortho, meta or para position.

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