Preparation method of novel thiourea substance, bacteriostatic action of composition and application of composition in promoting plant growth

Abstract

The invention discloses a novel thiourea substance, which is characterized in that the structural formula of the novel thiourea substance is as follows: # imgabs0 #, wherein R phenyl, substituted phenyl and heterocyclic ring. A biological activity determination result shows that the acylthiourea compound containing the purine part has good inhibitory activity on Xanthomonas oryzae pv. Oryzae, Xanthomonas citri pv. Citri and Xanthomonas syringae pv. Kiwifruit. The compound C19 has high activity, especially has medium to excellent inhibitory activity on rice bacterial leaf streak, the activity of the compound C19 is the best, and the inhibitory activity on the rice bacterial leaf streak reaches 92.0% and is far higher than that of control agents thiediazole copper and zinc thiazole.

Description

Technical Field

[0001] The invention relates to the field of chemical technology, in particular to a thiourea plant growth regulator, a preparation method of the compound and use of the composition thereof on rice bacterial leaf streak pathogen, kiwi fruit bacterial canker pathogen and citrus canker pathogen. Background Art

[0002] Repeated use of a single type of pesticide or pesticides with similar fungicidal mechanisms in large doses throughout the year not only wastes resources and aggravates environmental pollution, but also causes pests to develop resistance, the scope and intensity of which are expanding and increasing. Therefore, it is urgent to speed up the development of green pesticides with high efficiency, low toxicity, low residue and novel mechanism of action.

[0003] Thiourea derivatives have the potential for the development of this type of new pesticides. They all contain the group -HN-CS-NH-, and the chemical properties of this type of structure are active, thereby showing good biological activity. At the beginning of the last century, there were reports on the synthesis of thiourea derivatives. Ammonium thiocyanate was heated to 160 ° C, and after a period of reaction, it re-excluded the simplest thiourea-thiourea. After years of research and development, a large number of thiourea derivatives have been prepared, which have great potential use value in pesticides, medicines, supramolecules, chiral molecules, atmospheric pollutant monitoring, etc. For example: herbicides, bactericides, anti-tuberculosis drugs, anti-tumor drugs, antiviral drugs, regulating plant growth and development, chelating agents, organic synthesis catalysts, etc.

[0004] Pyrimidine compounds are widely present in organisms and are essential substances in life activities. For example, there are five nitrogenous bases in deoxynucleic acid (DNA) and ribonucleic acid (RNA), of which three have pyrimidine structures, namely cytosine (C), uracil (U) and thymine (T). Pyrimidine compounds can not only participate in various activities of the body, but also are an important intermediate in organic synthesis. They have attracted widespread attention because of their various biological activities, especially in the fields of medicine and pesticides. Pyrimidine compounds are new systemic drugs and are widely used as insecticides, fungicides, herbicides and plant growth regulators, such as pirimicarb, pirimiphos-methyl, pyrimethanil, azoxystrobin, etc.; not only that, pyrimidine compounds can also be used as intermediates for the synthesis of a new generation of pyrimidine pesticides. Therefore, their synthetic improvement has high application value and commercial value.

[0005] In summary, this paper introduces the active group of p-hydroxyaniline into halogenated pyrimidine through nucleophilic substitution and other reactions to synthesize pyrimidinyl aniline compounds with potential biological activity. In addition, novel pyrimidinyl thiourea compounds with potential biological activity are synthesized through the condensation reaction of pyrimidinyl aniline compounds with substituted isothiocyanates. The antibacterial and plant growth regulating activities are tested in order to seek efficient pesticide lead substances and screen out highly active antibacterial plant growth regulating drugs. Summary of the invention

[0006] The present invention aims to provide a method for preparing a thiourea biological plant growth regulator having bactericidal activity and a composition thereof.

[0007] Another object of the present invention is to provide a novel compound having an inhibitory effect on kiwi fruit bacterial canker, rice bacterial leaf streak pathogen and citrus canker pathogen.

[0008] The technical solution of the present invention is: a novel ferulic acid derivative containing a sulfonamide structure, wherein the derivative has the following general formula (I):

[0009]

[0010] Wherein: R is phenyl, 2-methylphenyl, 3-methylphenyl, 4-methylphenyl, 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 2-trifluoromethylphenyl, 3-trifluoromethylphenyl, 4-trifluoromethylphenyl, 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2-chlorophenyl, 3-chlorophenyl, 4-chlorophenyl, 2-bromophenyl, 3-bromophenyl, 4-bromophenyl, 4-iodophenyl, 4-biphenyl, 3,4-dimethoxyphenyl, 2,4-dichlorophenyl, 3,4-dichlorophenyl, 2-chloro-4-fluorophenyl, 1-naphthyl, 2-naphthyl or a disubstituted group of any combination of the above substituents.

[0011] Preferably, R is phenyl, 2-methylphenyl, 3-methylphenyl, 4-methylphenyl, 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 2-trifluoromethylphenyl, 3-trifluoromethylphenyl, 4-trifluoromethylphenyl, 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2-chlorophenyl, 3-chlorophenyl, 4-chlorophenyl, 2-bromophenyl, 3-bromophenyl, 4-bromophenyl, 4-iodophenyl, 4-biphenyl, 3,4-dimethoxyphenyl, 2,4-dichlorophenyl, 3,4-dichlorophenyl, 2-chloro-4-fluorophenyl, 1-naphthyl, 2-naphthyl or a disubstituted ...

[0012] A new type of thiourea substance, the specific compounds are as follows:

[0013] Compound C1: N-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)benzamide;

[0014] Compound C2: N-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-2-methylbenzamide;

[0015] Compound C3: N-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-3-methylbenzamide;

[0016] Compound C4: N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-4-methylbenzamide;

[0017] Compound C5: N-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-2-methoxybenzamide;

[0018] Compound C6: N-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-3-methoxybenzamide;

[0019] Compound C7: N-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-4-methoxybenzamide;

[0020] Compound C8: N-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-2-(trifluoromethyl)benzamide;

[0021] Compound C9: N-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-3-(trifluoromethyl)benzamide;

[0022] Compound C10: N-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-4-(trifluoromethyl)benzamide;

[0023] Compound C11: N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-2-fluorobenzamide;

[0024] Compound C12: N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-3-fluorobenzamide;

[0025] Compound C13: N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-4-fluorobenzamide;

[0026] Compound C14: 2-chloro-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)benzamide;

[0027] Compound C15: 3-chloro-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)benzamide;

[0028] Compound C16: 4-chloro-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)benzamide;

[0029] Compound C17: 2-bromo-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)benzamide;

[0030] Compound C18: 3-bromo-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)benzamide;

[0031] Compound C19: 4-bromo-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)benzamide;

[0032] Compound C20: N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-4-iodobenzamide;

[0033] Compound C21: N-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-[1,1′-biphenyl]-4-carboxamide.

[0034] Compound C22: N-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-3,4-dimethoxybenzamide.

[0035] Compound C23: 2,4-dichloro-N-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)benzamide.

[0036] Compound C24: 3,4-dichloro-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)benzamide.

[0037] Compound C25: 2-chloro-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-4-fluorobenzamide.

[0038] Compound C26: N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-1-naphthylamine.

[0039] Compound C27: N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamoyl)-2-naphthamide.

[0040] A method for preparing a purine derivative comprises the following steps:

[0041] (1) 6-chloro-9H-purine: bromomethylcyclopropane: potassium carbonate = 1:1.2:3, DMF 10mL, react at room temperature for 7h. After the reaction is completed, the mixture is poured into water and extracted three times with ethyl acetate to separate the organic phase. The organic layer is then washed with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product is purified by silica gel column chromatography using petroleum ether / ethyl acetate (1:1, v / v) as the eluent to obtain the key intermediate A;

[0042]

[0043] (2) Add p-aminophenol: intermediate A: potassium carbonate = 1:1:2, add 10 mL of acetonitrile, and react at 85°C for 7 hours. After the reaction is completed, add 40 mL of ethyl acetate to the reaction mixture, wash three times with water and saturated brine, respectively, dry in anhydrous Na2SO4 and concentrate. Use petroleum ether / ethyl acetate (1 / 1, v / v) as the eluent to purify the crude product by silica gel column chromatography to obtain a white solid compound B;

[0044]

[0045] (3) Various acyl isothiocyanates: intermediate amine B = 1:1.1 were added to THF 25 mL and reacted at room temperature for 6 hours. After the reaction was completed, the reaction solution was concentrated, poured into cold water, and extracted with dichloromethane three times. The obtained organic phase was dried over anhydrous Na2SO4, the solvent was removed by distillation under reduced pressure, and finally recrystallized from ethyl acetate to obtain the target compounds C1-C27;

[0046]

[0047] The derivative is used in preparing medicines and medicaments for preventing and controlling rice bacterial leaf streak pathogen, kiwi fruit bacterial canker pathogen and citrus canker pathogen.

[0048] A compound pesticide composition, characterized in that: the composition comprises the novel thiourea substance according to claim 1 and allicin. The mass ratio of the derivative to Zhongshengmycin is 1:2-2:1. The derivative is C19.

[0049] The composition is used in the preparation of a drug for preventing and treating plant bacterial diseases, wherein the plant bacterial disease is rice bacterial leaf streak pathogen.

[0050] A novel plant growth regulator is compound C19, a compound of the following formula (II):

[0051]

[0052] Beneficial effects of the present invention: From the results of biological activity assay, it can be seen that the acylthiourea compounds containing purine moieties have good inhibitory activity against rice bacterial leaf streak pathogen, citrus canker pathogen and kiwi fruit bacterial canker pathogen. In particular, they have moderate to excellent inhibitory activity against rice bacterial leaf streak pathogen, among which compound C19 has the best activity, and the inhibitory activity against rice bacterial leaf streak pathogen reaches 92.0%, which is much higher than the control agents thiophanate-copper and thiazole zinc.

[0053] From the results of biological activity determination, it can be seen that the activity of the composite composition against rice bacterial leaf streak pathogen is improved compared with the compound before compounding. The inhibitory activity of composition 3 (C19: allicin wettable powder = 2:1) against rice bacterial leaf streak pathogen is 98.7%. Therefore, the composite composition of C19 and allicin has a synergistic effect on rice bacterial leaf streak pathogen.

[0054] Plant growth activity of target compounds: Preliminary biological activity test results show that the target compounds have certain plant growth activity, among which compound C7 has good auxin activity, but it is less active than kinetin. Within a certain range, the activity of high concentration is better than that of low concentration, such as: the activity of compound C1 and compound C7 is slightly higher than that of indoleacetic acid, which can be further studied. DETAILED DESCRIPTION

[0055] The synthesis method of target compounds C1-C27 is as follows: Different substituted acyl chlorides are first reacted with potassium thiocyanate in acetone to obtain various acyl isothiocyanates. Using potassium carbonate as a catalyst, 6-chloro-9H-purine and bromomethylcyclopropane are reacted in DMF to obtain intermediate A. Intermediate A is coupled with p-aminophenol in acetonitrile, with K2CO3 as a catalyst, to generate key intermediate B. Finally, intermediate B is acylated with various acyl isothiocyanates in THF at room temperature to obtain target compounds C1-C27. All target compounds are prepared by 1 H NMR, 13The structure was confirmed by C NMR and HRMS.

[0056]

[0057] Preparation of intermediate B:

[0058] A mixture of p-aminophenol (3.0 mmol) and potassium carbonate (0.76 g, 5.51 mmol) was placed in an acetonitrile solution and stirred at room temperature for 0.5 h. Then, intermediate A (3.0 mmol) was added to the above solution and refluxed at 85°C for 7 h. After the reaction was completed, 40 mL of ethyl acetate was added to the reaction mixture, which was washed three times with water and saturated brine, dried in anhydrous Na2SO4 and concentrated. The crude product was purified by silica gel column chromatography using petroleum ether / ethyl acetate (1 / 1, v / v) as the eluent to obtain white solid compound B.

[0059] Preparation of target compounds C1-C27

[0060] Various acyl isothiocyanates (2.0 mmol) were slowly added to the intermediate amine B (0.56 g, 2.2 mmol) dissolved in THF (25 mL) and stirred at room temperature for 6 h. After the reaction was completed (TLC monitoring), the reaction solution was concentrated, poured into cold water, and extracted three times with dichloromethane. The obtained organic phase was dried over anhydrous Na2SO4, the solvent was removed by distillation under reduced pressure, and finally recrystallized in ethyl acetate to obtain the target compound C1-C27.

[0061] Spectral data of target compounds

[0062]

[0063] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)benzamide(C1): white solid, mp 194–195℃, yield: 53.6%. 1 H NMR (400 MHz, DMSO-d6, ppm) δ: 12.62 (s, 1H), 11.62 (s, 1H), 8.60 (s, 1H), 8.49 (s, 1H), 8.00 (d, J = 7.0 Hz, 2H), 7.78 (d, J = 8.9 Hz, 2H), 7.67 (t, J = 7.4 Hz,1H),7.55(t,J=7.7 Hz,2H),7.39–7.35(m,2H),4.15(d,J=7.3 Hz,2H),1.36(hept,J=7.5 Hz,1H),0.57–0.46(m,4H); 13C NMR(100 MHz,DMSO-d6,ppm)δ:179.4,168.3,159.3,153.2,151.2,150.1,144.8,135.4,133.2,132.2,128.7,128.5,125.8,122.1,120.7,47.9,11.3,3.9.HRMS(ESI)m / z:[M+Na] + calcd forC 23 H 20 N6O2SNa:467.1261,found:467.1261.

[0064]

[0065] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-2-methylbenzamide(C2):white solid,mp 171–173℃,yield:55.4%. 1 HNMR(400 MHz,DMSO-d6,ppm)δ:12.55(s,1H),11.75(s,1H),8.60(s,1H),8.49(s,1H),7.81–7.78(m,2H),7.53(d,J=7.2 Hz,1H),7.45(t,J=8.3 Hz,1H),7.38–7.29(m,4H),4.16(d,J=7.3 Hz,2H),2.44(s,3H),1.36(hept,J=7.6 Hz,1H),0.58–0.46(m,4H); 13 CNMR(100MHz,DMSO-d6,ppm)δ:179.2,170.5,159.3,153.2,151.2,150.1,144.8,136.1,135.3,134.0,131.0,130.6,128.2,125.7,125.5,122.1,120.7,47.9,19.5,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd for C 24 H 22 N6O2SNa:481.1417,found:481.1422.

[0066]

[0067] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-3-methylbenzamide(C3):white solid,mp 195–196℃,yield:60.5%. 1 HNMR(400 MHz,DMSO-d6,ppm)δ:12.64(s,1H),11.55(s,1H),8.61(s,1H),8.48(s,1H),7.85(s,1H),7.81–7.77(m,3H),7.49–7.41(m,2H),7.38–7.34(m,2H),4.15(d,J=7.3Hz,2H),2.40(s,3H),1.35(hept,J=7.5 Hz,1H),0.57–0.46(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:179.3,168.3,159.3,153.2,151.2,150.1,144.8,137.9,135.3,133.8,132.0,129.1,128.4,125.9,125.8,122.1,120.7,47.9,20.8,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd for C 24 H 22 N6O2SNa:481.1417,found:481.1412.

[0068]

[0069] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-4-methylbenzamide(C4):white solid,mp 195–196℃,yield:62.8%. 1HNMR(400 MHz,DMSO-d6,ppm)δ:12.67(s,1H),11.51(s,1H),8.60(s,1H),8.48(s,1H),7.93(d,J=8.0 Hz,2H),7.79–7.76(m,2H),7.38–7.34(m,4H),4.15(d,J=7.3 Hz,2H),2.40(s,3H),1.36(hept,J=7.6 Hz,1H),0.57–0.46(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:179.4,168.1,159.3,153.2,151.2,150.1,144.8,143.7,135.4,129.2,129.1,128.8,125.8,122.1,120.7,47.9,21.2,11.3,3.9.HRMS(ESI)m / z:[M+Na] + calcd forC 24 H 22 N6O2SNa:481.1417,found:481.1433.

[0070]

[0071] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-2-methoxybenzamide(C5):white solid,mp 182–184℃,yield:53.7%. 1 H NMR(400 MHz,DMSO-d6,ppm)δ:12.56(s,1H),11.26(s,1H),8.60(s,1H),8.48(s,1H),7.94(dd,J=7.8,1.8 Hz,1H),7.81–7.77(m,2H),7.67(t,J=8.8 Hz,1H),7.39–7.35(m,2H),7.30(d,J=8.4 Hz,1H),7.17(t,J=7.4 Hz,1H),4.15(d,J=7.3 Hz,2H),4.03(s,3H),1.36(hept,J=7.5 Hz,1H),0.57–0.46(m,4H); 13C NMR(100 MHz,DMSO-d6,ppm)δ:178.3 165.3,159.3,157.6,153.2,151.2,150.2,144.8,135.2,135.1,131.2,125.7,122.2,121.3,120.7,119.5,112.9,56.7,47.9,11.3,3.9.HRMS(ESI)m / z:[M+Na] + calcdfor C 24 H 22 N6O3SNa:497.1366,found:497.1380.

[0072]

[0073] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-3-methoxybenzamide(C6):white solid,mp 192–193℃,yield:59.6%. 1 H NMR(400 MHz,DMSO-d6,ppm)δ:12.62(s,1H),11.62(s,1H),8.60(s,1H),8.49(s,1H),7.78(d,J=8.4 Hz,2H),7.60–7.56(m,2H),7.46(t,J=7.9 Hz,1H),7.38–7.35(m,2H),7.22(dd,J=8.3,2.6 Hz,1H),4.15(d,J=7.3 Hz,2H),3.86(s,3H),1.35(hept,J=7.5Hz,1H),0.57–0.46(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:179.3,167.9,159.3,159.0,153.2,151.2,150.1,144.8,135.3,133.4,129.7,125.8,122.1,121.0,120.7,119.5,113.3,55.5,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd forC 24 H 22 N6O3SNa:497.1366,found:497.1367.

[0074]

[0075] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-4-methoxybenzamide(C7):white solid,mp 191–192℃,yield:64.3%. 1 H NMR(400 MHz,DMSO-d6,ppm)δ:12.73(s,1H),11.43(s,1H),8.60(s,1H),8.48(s,1H),8.06–8.02(m,2H),7.78–7.75(m,2H),7.38–7.34(m,2H),7.10–7.06(m,2H),4.15(d,J=7.3Hz,2H),3.86(s,3H),1.35(hept,J=7.6 Hz,1H),0.57–0.46(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:179.5,167.5,163.3,159.3,153.2,151.2,150.0,144.7,135.4,131.0,125.8,123.8,122.1,120.7,113.8,55.6,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcdfor C 24 H 22 N6O3SNa:497.1366,found:497.1372.

[0076]

[0077] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-2-(trifluoromethyl)benzamide(C8):white solid,mp 185–186℃,yield:60.2%. 1H NMR(400 MHz,DMSO-d6,ppm)δ:12.29(s,1H),12.15(s,1H),8.60(s,1H),8.49(s,1H),7.87(d,J=7.6 Hz,1H),7.83–7.74(m,5H),7.39–7.35(m,2H),4.16(d,J=7.3Hz,2H),1.36(hept,J=7.5 Hz,1H),0.58–0.46(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:178.8,168.5,159.2,153.2,151.2,150.2,144.8,135.1,133.3(q, 4 J C-F =2.0Hz),132.5,131.0,128.9,126.3(q, 3 J C-F =5.0 Hz),125.9,125.8(q, 2 J C-F =31.0 Hz),123.6(q, 1 J C-F =272.0 Hz),122.1,120.7,47.92,11.32,3.84.HRMS(ESI)m / z:[M+Na] + calcd forC 24 H 19 F3N6O2SNa:535.1134,found:535.1135.

[0078]

[0079] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-3-(trifluoromethyl)benzamide(C9):white solid,mp 190–191℃,yield:62.3%. 1H NMR(400 MHz,DMSO-d6,ppm)δ:12.50(s,1H),11.97(s,1H),8.60(s,1H),8.49(s,1H),8.36(s,1H),8.26(d,J=8.0 Hz,1H),8.03(d,J=7.9 Hz,1H),7.81–7.76(m,3H),7.39–7.35(m,2H),4.15(d,J=7.3 Hz,2H),1.36(hept,J=7.6 Hz,1H),0.57–0.46(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:179.2,166.9,159.3,153.2,151.2,150.2,144.8,135.3,133.3,132.9,129.7,129.5,129.0(q, 2 J C-F =32.0 Hz),125.8,125.5(q, 3 J C-F =4.0Hz),123.8(q, 1 J C-F =271.0 Hz),122.1,120.7,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd for C 24 H 19 F3N6O2SNa:535.1134,found:535.1140.

[0080]

[0081] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-4-(trifluoromethyl)benzamide(C10):white solid,mp 211–212℃,yield:68.6%. 1H NMR(400 MHz,DMSO-d6,ppm)δ:12.46(s,1H),11.89(s,1H),8.60(s,1H),8.49(s,1H),8.17(d,J=8.1 Hz,2H),7.92(d,J=8.3 Hz,2H),7.79–7.76(m,2H),7.39–7.35(m,2H),4.15(d,J=7.3 Hz,2H),1.36(hept,J=7.5 Hz,1H),0.57–0.46(m,4H); 13 CNMR(150 MHz,DMSO-d6,ppm)δ:179.1,167.2,159.3,153.2,151.2,150.2,144.8,136.3,135.3,132.4(q, 2 J C-F =33.0 Hz),129.7,125.9,125.4(q, 3 J C-F =4.5 Hz),123.8(q, 1 J C-F =271.5Hz),122.2,120.7,47.9,11.3,3.9.HRMS(ESI)m / z:[M+Na] + calcdfor C 24 H 19 F3N6O2SNa:535.1134,found:535.1139.

[0082]

[0083] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-2-fluorobenzamide(C11):white solid,mp 193–194℃,yield:59.0%. 1 H NMR(400 MHz,DMSO-d6,ppm)δ:12.36(s,1H),11.75(s,1H),8.60(s,1H),8.49(s,1H),7.79–7.63(m,4H),7.40–7.34(m,4H),4.15(d,J=7.3 Hz,2H),1.36(hept,J=7.5 Hz,1H),0.58–0.46(m,4H); 13C NMR(150 MHz,DMSO-d6,ppm)δ:178.8,165.3,159.3(d, 1 J C-F =249.0Hz),159.2,153.2,151.2,150.2,144.8,135.2,134.2(d, 3 J C-F =8.0 Hz),130.4(d, 4 J C-F =2.0 Hz),125.9,124.6(d, 3 J C-F =4.0 Hz),122.3,122.1,120.7,116.2(d, 2 J C-F =21.0 Hz),47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd forC 23 H 19 FN6O2SNa:485.1166,found:485.1169.

[0084]

[0085] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-3-fluorobenzamide(C12):white solid,mp 196–197℃,yield:61.8%. 1 H NMR(400 MHz,DMSO-d6,ppm)δ:12.49(s,1H),11.71(s,1H),8.60(s,1H),8.48(s,1H),7.86–7.81(m,2H),7.78–7.73(m,2H),7.60(td,J=8.0,5.8 Hz,1H),7.52(t,J=7.7 Hz,1H),7.39–7.35(m,2H),4.15(d,J=7.3 Hz,2H),1.36(hept,J=7.6 Hz,1H),0.57–0.46(m,4H); 13 C NMR(150 MHz,DMSO-d6,ppm)δ:179.2,166.8,161.7(d, 1 J C-F=243.0 Hz),159.3,153.2,151.2,150.1,144.8,135.3,134.5(d, 3 J C-F =7.5 Hz),130.7(d, 3 J C-F =7.5 Hz),125.9,125.0(d, 4 J C-F =3.0 Hz),122.1,120.7,120.0(d, 2 J C-F =21.0Hz),115.6(d, 2 J C-F =24.0 Hz),47.9,11.3,3.9.HRMS(ESI)m / z:[M+Na] + calcd forC 23 H 19 FN6O2SNa:485.1166,found:485.1169.

[0086]

[0087] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-4-fluorobenzamide(C13):white solid,mp 184–185℃,yield:64.2%. 1 H NMR(400 MHz,DMSO-d6,ppm)δ:12.56(s,1H),11.67(s,1H),8.60(s,1H),8.48(s,1H),8.09(dd,J=8.8,5.6 Hz,2H),7.77(d,J=8.5 Hz,2H),7.41–7.35(m,4H),4.15(d,J=7.3Hz,2H),1.36(hept,J=7.6 Hz,1H),0.57–0.46(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:179.3,167.1,165.0(d, 1 J C-F =250.0 Hz),159.3,153.2,151.2,150.1,144.8,135.3,131.7(d, 3 J C-F =9.0 Hz),128.7(d, 4J C-F =3.0 Hz),125.8,122.1,120.7,115.5(d, 2 J C-F =22.0Hz),47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd forC 23 H 19 FN6O2SNa:485.1166,found:485.1163.

[0088]

[0089] 2-chloro-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)benzamide(C14):white solid,mp 193–194℃,yield:57.9%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:12.36(s,1H),12.05(s,1H),8.60(s,1H),8.49(s,1H),7.80–7.76(m,2H),7.66(dd,J=7.6,1.5 Hz,1H),7.59–7.53(m,2H),7.47(td,J=7.2,6.8,2.0Hz,1H),7.39–7.35(m,2H),4.15(d,J=7.3 Hz,2H),1.36(hept,J=7.5 Hz,1H),0.57–0.46(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:178.9,167.7,159.2,153.2,151.2,150.2,144.8,135.2,134.3,132.1,130.0,129.6,129.4,127.1,125.9,122.1,120.7,47.9,11.3,3.9.HRMS(ESI)m / z:[M+Na] + calcd forC 23 H 19 ClN6O2SNa:501.0871,found:501.0875.

[0090]

[0091] 3-chloro-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)benzamide(C15):white solid,mp 160–161℃,yield:60.6%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:12.48(s,1H),11.77(s,1H),8.60(s,1H),8.49(s,1H),8.05(s,1H),7.93(d,J=7.9 Hz,1H),7.78–7.75(m,2H),7.59(d,J=7.9 Hz,1H),7.49(t,J=7.9 Hz,1H),7.39–7.35(m,2H),4.15(d,J=7.3 Hz,2H),1.36(hept,J=7.6 Hz,1H),0.57–0.46(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:179.2,166.4,159.3,153.2,151.2,150.1,144.8,136.3,134.3,133.1,131.1,130.4,128.5,127.4,125.8,122.1,120.7,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd forC 23 H 19 ClN6O2SNa:501.0871,found:501.0871.

[0092]

[0093] 4-chloro-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)benzamide(C16):white solid,mp 197–198℃,yield:61.7%. 1H NMR(400MHz,DMSO-d6,ppm)δ:12.52(s,1H),11.72(s,1H),8.60(s,1H),8.48(s,1H),8.02–8.00(m,2H),7.77(d,J=8.6 Hz,2H),7.64–7.60(m,2H),7.38–7.34(m,2H),4.15(d,J=7.2 Hz,2H),1.35(hept,J=7.6 Hz,1H),0.57–0.46(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:179.2,167.2,159.3,153.2,151.2,150.1,144.8,138.0,135.3,131.0,130.7,128.5,125.8,122.1,120.7,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd for C 23 H 19 ClN6O2SNa:501.0871,found:501.0866.

[0094]

[0095] 2-bromo-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)benzamide(C17):white solid,mp 180–181℃,yield:56.9%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:12.37(s,1H),12.05(s,1H),8.60(s,1H),8.49(s,1H),7.81–7.78(m,2H),7.73(dd,J=7.7,1.4 Hz,1H),7.63(dd,J=7.4,1.9 Hz,1H),7.53–7.44(m,2H),7.39–7.35(m,2H),4.16(d,J=7.3 Hz,2H),1.36(hept,J=7.5 Hz,1H),0.58–0.46(m,4H); 13CNMR(150 MHz,DMSO-d6,ppm)δ:178.9,168.6,159.3,153.2,151.2,150.2,144.8,136.5,135.2,132.7,132.2,129.3,127.6,125.9,122.1,120.7,118.9,47.9,11.3,3.9.HRMS(ESI)m / z:[M+Na] + calcd for C 23 H 19 BrN6O2SNa:545.0366,found:545.0368.

[0096]

[0097] 3-bromo-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)benzamide(C18):white solid,mp 198–199℃,yield:59.6%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:12.48(s,1H),11.78(s,1H),8.60(s,1H),8.48(s,1H),8.18(s,1H),7.97(d,J=8.0 Hz,1H),7.86(d,J=8.0 Hz,1H),7.78–7.75(m,2H),7.51(t,J=7.9 Hz,1H),7.38–7.35(m,2H),4.15(d,J=7.3 Hz,2H),1.36(hept,J=7.6 Hz,1H),0.57–0.46(m,4H); 13 C NMR(150 MHz,DMSO-d6,ppm)δ:179.2,166.8,159.3,153.2,151.2,150.1,144.8,135.7,135.3,134.4,131.3,130.6,127.8,125.8,122.1,121.5,120.7,47.9,11.3,3.9.HRMS(ESI)m / z:[M+Na] + calcd forC 23 H 19 BrN6O2SNa:545.0366,found:545.0370.

[0098]

[0099] 4-bromo-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)benzamide(C19):white solid,mp 209–210℃,yield:60.3%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:12.51(s,1H),11.72(s,1H),8.60(s,1H),8.48(s,1H),7.95–7.91(m,2H),7.78–7.74(m,4H),7.38–7.34(m,2H),4.15(d,J=7.3 Hz,2H),1.35(hept,J=7.6 Hz,1H),0.57–0.46(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:179.2,167.3,159.3,153.2,151.2,150.1,144.8,135.3,131.5,131.4,130.8,127.1,125.8,122.1,120.7,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd forC 23 H 19 BrN6O2SNa:545.0366,found:545.0367.

[0100]

[0101] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-4-iodobenzamide(C20):white solid,mp 216–217℃,yield:48.5%. 1 HNMR(400 MHz,DMSO-d6,ppm)δ:12.52(s,1H),11.69(s,1H),8.60(s,1H),8.48(s,1H),7.96–7.92(m,2H),7.77–7.75(m,4H),7.38–7.34(m,2H),4.15(d,J=7.3 Hz,2H),1.36(hept,J=7.6 Hz,1H),0.57–0.46(m,4H); 13C NMR(100 MHz,DMSO-d6,ppm)δ:179.2,167.7,159.3,153.2,151.2,150.1,144.8,137.3,135.3,131.6,130.5,125.8,122.1,120.7,101.6,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd forC 23 H 19 IN6O2SNa:593.0227,found:593.0226.

[0102]

[0103] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-[1,1'-biphenyl]-4-carboxamide(C21):white solid,mp 210–211℃,yield:64.2%. 1 H NMR(400 MHz,DMSO-d6,ppm)δ:12.67(s,1H),11.66(s,1H),8.60(s,1H),8.49(s,1H),8.13–8.10(m,2H),7.87–7.84(m,2H),7.81–7.75(m,4H),7.52(t,J=7.4 Hz,2H),7.44(t,J=7.3 Hz,1H),7.39–7.35(m,2H),4.15(d,J=7.3 Hz,2H),1.36(hept,J=7.5 Hz,1H),0.57–0.46(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:179.3,167.9,159.3,153.2,151.2,150.1,144.8,144.6,138.7,135.3,130.9,129.5,129.1,128.5,127.0,126.6,125.8,122.1,120.7,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd forC 29 H 24 N6O2SNa:543.1574,found:543.1568.

[0104]

[0105] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-3,4-dimethoxybenzamide(C22):white solid,mp 200–201℃,yield:58.9%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:12.79(s,1H),11.48(s,1H),8.60(s,1H),8.48(s,1H),7.78–7.71(m,3H),7.65(d,J=2.2 Hz,1H),7.38–7.34(m,2H),7.11(d,J=8.7 Hz,1H),4.15(d,J=7.3 Hz,2H),3.87(d,J=5.3 Hz,6H),1.35(hept,J=7.5 Hz,1H),0.57–0.46(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:179.5,167.5,159.3,153.2,153.1,151.2,150.1,148.2,144.8,135.4,125.8,123.7,123.0,122.1,120.7,111.5,111.0,55.8,55.7,47.9,11.3,3.9.HRMS(ESI)m / z:[M+Na] + calcd forC 25 H 24 N6O4SNa:527.1472,found:527.1476.

[0106]

[0107] 2,4-dichloro-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)benzamide(C23):white solid,mp 197–198℃,yield:61.3%. 1H NMR(400MHz,DMSO-d6,ppm)δ:12.27(s,1H),12.07(s,1H),8.60(s,1H),8.49(s,1H),7.78–7.75(m,3H),7.70(d,J=8.3 Hz,1H),7.57(dd,J=8.3,2.0 Hz,1H),7.39–7.35(m,2H),4.15(d,J=7.3 Hz,2H),1.36(hept,J=7.6 Hz,1H),0.57–0.46(m,4H); 13 C NMR(150 MHz,DMSO-d6,ppm)δ:178.8,166.8,159.3,153.2,151.2,150.2,144.8,136.0,135.1,133.3,131.4,130.8,129.2,127.4,125.9,122.1,120.7,47.9,11.3,3.9.HRMS(ESI)m / z:[M+Na] + calcdfor C 23 H 18 Cl2N6O2SNa:535.0481,found:535.0488.

[0108]

[0109] 3,4-dichloro-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)benzamide(C24):white solid,mp 197–198℃,yield:57.9%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:12.42(s,1H),11.83(s,1H),8.59(s,1H),8.48(s,1H),8.26(d,J=2.1 Hz,1H),7.94(dd,J=8.4,2.1 Hz,1H),7.82(d,J=8.4 Hz,1H),7.77–7.74(m,2H),7.38–7.34(m,2H),4.15(d,J=7.3 Hz,2H),1.35(hept,J=7.6 Hz,1H),0.57–0.46(m,4H); 13C NMR(100 MHz,DMSO-d6,ppm)δ:179.1,166.0,159.3,153.2,151.2,150.2,144.8,135.9,135.3,132.7,131.3,130.8,130.7,129.0,125.8,122.1,120.7,47.9,11.3,3.9.HRMS(ESI)m / z:[M+Na] + calcd forC 23 H 18 Cl2N6O2SNa:535.0481,found:535.0476.

[0110]

[0111] 2-chloro-N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-4-fluorobenzamide(C25):white solid,mp 202–203℃,yield:60.9%. 1 H NMR(400 MHz,DMSO-d6,ppm)δ:12.31(s,1H),12.04(s,1H),8.60(s,1H),8.49(s,1H),7.79–7.73(m,3H),7.61(dd,J=9.0,2.5 Hz,1H),7.39–7.34(m,3H),4.15(d,J=7.3 Hz,2H),1.36(hept,J=7.6 Hz,1H),0.57–0.46(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:178.8,166.9,162.9(d, 1 J C-F =250.0 Hz),159.2,153.2,151.2,150.2,144.8,135.1,131.7(d, 3 J C-F =11.0 Hz),131.5(d, 3 J C-F =9.0 Hz),131.0(d, 4 J C-F =3.0 Hz),125.9,122.1,120.7,117.1(d, 2 J C-F =26.0 Hz),114.5(d,2 J C-F =21.0Hz),47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd for C 23 H 18 ClFN6O2SNa:519.0777,found:519.0773.

[0112]

[0113] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-1-naphthamide(C26):white solid,mp 210–211℃,yield:64.7%. 1 HNMR(400 MHz,DMSO-d6,ppm)δ:12.64(s,1H),12.03(s,1H),8.61(s,1H),8.51(s,1H),8.26(d,J=8.3 Hz,1H),8.14(d,J=8.4 Hz,1H),8.05(dd,J=8.3,1.4 Hz,1H),7.87–7.83(m,3H),7.69–7.60(m,3H),7.41–7.37(m,2H),4.16(d,J=7.3 Hz,2H),1.36(hept,J=7.6 Hz,1H),0.58–0.47(m,4H); 13 C NMR(100 MHz,DMSO-d6,ppm)δ:179.3,169.9,159.3,153.2,151.2,150.1,144.8,135.3,133.0,131.6,131.4,129.5,128.5,127.4,127.2,126.5,125.8,124.8,124.7,122.1,120.7,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd forC 27 H 22 N6O2SNa:517.1417,found:517.1419.

[0114]

[0115] N-((4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)phenyl)carbamothioyl)-2-napht hamide(C27): white solid, mp 205–206℃, yield: 63.1%. 1 HNMR(400MHz,DMSO-d6,ppm)δ:12.68(s,1H),11.78(s,1H),8.74(s,1H),8.60(s,1H),8.49(s,1H),8.13(d,J=8.0Hz,1H),8.08–8.00 (m,3H),7.82–7.80(m,2H),7.72–7.63(m,2H),7.40–7.36(m,2H),4.15(d,J=7.3Hz,2H),1.36(hept,J=7.6Hz,1H),0.57–0.46(m,4H); 13 CNMR(100MHz,DMSO-d6,ppm)δ:179.4,168.3,159.3,153.2,151.2,150.1,144.8,135.4,134.9,131.7,130.1, 129.4,129.2,128.7,128.1,127.7,127.1,125.8,124.5,122.1,120.7,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + Calculate for C 27 H 22 N6O2SNa:517.1417,found:517.1413.

[0116] 3. Antimicrobial activity data of target compounds

[0117] Test method: The in vitro antibacterial effect of the target compounds C1-C27 on three plant pathogens (Xoc, Psa, Xac) was determined by the classical turbidity method. The test compounds were prepared at two concentrations of 100 and 50 μg / mL. The blank control was pure dimethyl sulfoxide in sterile distilled water, and the positive control agents were commercially available fungicides zinc thiazole and thiophanate-methyl. About 40 μL NB solvent (3.0 g beef extract, 5.0 g peptone, 1.0 g yeast powder, 10.0 g glucose, 1000 mL distilled water, pH 7.0-7.2) containing bacteria Xoc, Xac or Psa was added to a mixed solvent system containing 4 mL NB solvent and 1 mL 0.1% Tween-20 (containing the test compound or BMT). The test tubes were incubated at 28±1°C and shaken continuously at 180 rpm for 1 to 2 days. The optical density at 595 nm (OD 595 ) to monitor bacterial growth.

[0118] I=(C tur -T tur ) / C tur ×100%

[0119] Among them, C tur is the corrected turbidity value of bacterial growth in untreated NB (blank control), T tur is the corrected turbidity value for bacterial growth treated with the test compound. Finally, I represents the inhibition rate of the test compound on bacteria.

[0120] Table 1. In vitro antibacterial activity of compounds C1-C27 against Xanthomonas oryzae, Xanthomonas syringae and Xanthomonas citri

[0121]

[0122] From the results of biological activity assay in Table 1, it can be seen that the acylthiourea compounds containing purine moieties have good inhibitory activity against rice bacterial leaf streak pathogen, citrus canker pathogen and kiwi fruit bacterial canker pathogen. In particular, they have moderate to excellent inhibitory activity against rice bacterial leaf streak pathogen, among which compound C19 has the best activity, with an inhibitory activity against rice bacterial leaf streak pathogen reaching 92.0%, which is much higher than the control agents thiophanate-copper and thiazole zinc.

[0123] 4. Preparation of target compound C19 composition

[0124] The wettable powder of the target compound C19 and allicin is used to prepare a composite composition. In each composition, the ratio of the target compound C19 and allicin is calculated according to the mass ratio. The following preparations of the target compound C19 and allicin are prepared as needed.

[0125] Composition 1: C19: Allicin wettable powder = 1:1

[0126] Composition 2: C19: Allicin wettable powder = 1:2

[0127] Composition 3: C19: Allicin wettable powder = 2:1

[0128] Anti-rice bacterial leaf streak activity of the composition of target compound C19

[0129] The antibacterial activity of the combination of target compound C19 and allicin against oryzae oryzae was tested according to the turbidity method (the antibacterial activity test method mentioned previously).

[0130] Table 2 Inhibitory activity of drugs against bacterial leaf streak pathogen of rice

[0131]

[0132] The in vitro growth rate method was used to test the activity of the composition against rice bacterial leaf streak pathogen at a concentration of 100 μg / mL. From the biological activity test results in Table 2, it can be seen that the activity of the composite composition against rice bacterial leaf streak pathogen is improved compared with the compounds before compounding. The inhibitory activity of composition 3 (C19: allicin wettable powder = 2:1) against rice bacterial leaf streak pathogen is 98.7%. Therefore, the composite composition of C19 and allicin has a synergistic effect on rice bacterial leaf streak pathogen.

[0133] 5. Determination of plant growth regulating activity of some target compounds

[0134] (1) Test method

[0135] The wheat sheath cutting test method and the radish cotyledon expansion method were used. The plant growth regulating activity of the target compound was determined. Preparation of buffered nutrient solution: Weigh 1.019g of citric acid, 2.350g of potassium hydrogen phosphate trihydrate, and 20g of sugar, mix and dilute to 1L, and a citric acid-phosphate buffer solution with a pH of -5 is obtained. In the wheat sheath test method, indoleacetic acid (IAA) is used as the standard control, and in the radish cotyledon expansion method, kinetin (KT) is used as the standard control. Preparation of sample solution and standard control solution: Weigh an appropriate amount of the target compound and the control sample, add a few drops of DMF to dissolve, then add - drops of emulsifier (Tween-80), and use the buffer solution to prepare sample solutions with a concentration of 100μg / mL, and then dilute to 50, 10, and 1μg / mL in sequence. Preparation of germination bed: Weigh 2.4g of agar, heat 400mL of water until dissolved, pour into a porcelain plate and cool to form a germination bed.

[0136] (1) Wheat bud sheath cutting test method:

[0137] First, weigh about 50g of wheat seeds, soak them for 4 hours, then rinse them repeatedly with clean water, drain the water, evenly place them in the germination bed, seal them with a film, and then place them in an artificial climate box at a temperature of 25±1℃ for cultivation in a dark environment. When the wheat bud sheath grows to 2.5-3cm, select the bud sheaths with the same growth trend, use a blade to cut out their sensitive sections, and only take one section from each bud sheath. After cutting, shade them and rinse them in distilled water for one hour. Then, measure 8mL of each sample solution and place them in a culture dish with a diameter of 9cm. Two filter paper sheets are placed in each culture dish. Then, the cut 10 bud sheath segments are evenly placed in each treated culture dish, and then placed in an artificial climate box at 25±1℃ for dark cultivation again. After 48 hours, measure the total length of the 10 bud sheaths, compare them with the blank buffer solution control group, and compare them with the standard solution control group of the same concentration. The inhibitory or promoting effect is calculated and used as an evaluation index for the function of drug auxin. Calculate according to the following formula:

[0138] Effect = (treatment - blank) / blank × 100%

[0139] If the calculation result is positive, it means there is a promoting effect; if the calculation result is negative, it means there is an inhibitory effect. Finally, the efficacy is evaluated. The evaluation criteria are:

[0140] 40% ≤ Effect Grade A

[0141] 25% ≤ Effect < 40% Grade B

[0142] 10% ≤ effect < 25% Grade C

[0143] Effect <10% D

[0144] (2) Radish cotyledon expansion test method:

[0145] First weigh about 30g of radish seeds, soak them in warm water until the seeds turn white, then wash them with clean water, drain the water and put them in the germination bed, seal them with a film, and then place them in an artificial climate box at 25±1℃ for dark culture for about 72 hours, and set aside after completion. Measure 8mL of each sample solution and place it in a culture dish with a diameter of 9cm, and put two pieces of paper in each culture dish. Select radish seedlings with uniform growth and cotyledon size, cut off the cotyledons without petioles, place 10 cotyledons for each treatment, and weigh the fresh weight of the cotyledons before treatment in each group and record the data, then cover the culture dish cover and place it in an artificial climate box for dark culture. After 72 hours, take out the cotyledons of each group, use absorbent paper to absorb the moisture on the surface of the cotyledons, weigh the fresh weight of the cotyledons after treatment in each group and record the data, calculate the percentage of fresh weight gain of the cotyledons after culture, and use this as the evaluation index of the function of the drug sample cytokinin. Its calculation method and evaluation criteria are the same as those of the wheat bud sheath cutting test method.

[0146] (3) Biological test results

[0147] Table 3 Plant growth regulator activity of some target compounds

[0148]

[0149] The plant growth activity test results of this series of target compounds are shown in Table 3. The preliminary biological activity test results show that the target compounds have certain plant growth activity, among which compound C7 has good auxin activity, but it is less active than kinetin. Within a certain range, the activity of high concentration is better than that of low concentration, such as: the activity of compound C1 and compound C7 is slightly higher than that of indoleacetic acid, which can be further studied.

Claims

1. A novel thiourea substance, characterized in that: The novel thiourea substance has the following structural formula: , wherein R is phenyl, substituted phenyl, or heterocycle.

2. A novel thiourea substance according to claim 1, characterized in that: The R is phenyl, substituted phenyl, or heterocyclic ring, the substituent of the substituted phenyl is methyl, methoxy, trifluoromethyl, or halogen, and the heterocyclic ring is naphthalene.

3. The method for preparing a novel thiourea substance according to claim 1, characterized in that: The reaction formula is as follows:

4. The method for preparing a novel thiourea substance according to claim 3, characterized in that: The method comprises the following steps: firstly, acyl chlorides with different substitutions react with potassium thiocyanate in acetone to obtain various acyl isothiocyanates; with potassium carbonate as a catalyst, 6-chloro-9H-purine reacts with bromomethylcyclopropane in DMF to obtain an intermediate A; intermediate A is coupled with p-aminophenol in acetonitrile with K2CO3 as a catalyst to generate a key intermediate B; finally, intermediate B is subjected to an acylation reaction with various acyl isothiocyanates in THF at room temperature to obtain a target compound.

5. Use of a novel thiourea substance as claimed in claim 1 or 2 in the preparation of drugs and medicaments for preventing and controlling bacterial leaf streak pathogen of rice, bacterial canker pathogen of kiwi fruit and bacterial canker pathogen of citrus.

6. A compound pesticide composition, characterized in that: The composition comprises the novel thiourea substance according to claim 1 and allicin.

7. The composition according to claim 6, characterized in that: The mass ratio of the novel thiourea substance to allicin is 1:2-2:

1.

8. Use of the composition according to claim 6 in the preparation of a medicament for controlling bacterial leaf streak pathogen of rice.

9. The composition according to claim 6, characterized in that: The derivative is C19.

10. Application of the novel thiourea substance as claimed in claim 1 or 2 in the preparation of plant growth regulators.

Images