Purine plant growth regulating substance, preparation method and application of composition of purine plant growth regulating substance

Abstract

The invention discloses a purine plant growth regulating substance, which is characterized in that the structural formula of the purine plant growth regulating substance is as follows: # imgabs0 #, in which R1 is C3-C4 alkyl and cycloalkyl, R2 is phenyl, substituted phenyl and heterocyclic radical. A biological activity determination result of the compound shows that the acylhydrazone compound containing the purine part has medium to excellent inhibitory activity on rice bacterial leaf streak germs and citrus canker germs, the activity of the compound A7 is the best, and the inhibitory activity on the rice bacterial leaf streak germs and the citrus canker germs is 77.6% and 54.8% respectively; the content is 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 purine plant growth regulator, a preparation method of the compound and use of the composition thereof on kiwifruit bacterial canker, rice bacterial leaf streak pathogen and citrus canker pathogen. Background Art

[0002] Plant diseases caused by bacteria lead to a significant decrease in grain yields of crops. Diseases caused by pathogens such as rice bacterial leaf streak fungus, citrus canker and kiwi bacterial canker have caused huge economic losses to agriculture. At present, the commonly used antibacterial agents on the market not only have poor field prevention effects and are not conducive to environmental sustainable development, but also increase bacterial resistance. Therefore, it is urgent to develop an antibacterial agent that is highly efficient, low-toxic and environmentally friendly.

[0003] Plant hormones refer to trace organic substances naturally present in plants that have significant effects on plant growth and development. They are also called plant natural hormones or plant endogenous hormones. Their presence can affect and effectively regulate plant growth and development, including a series of plant life processes from cell growth and division to rooting, germination, flowering, fruiting, maturity and shedding. Plant growth regulators are a class of substances with similar physiological and biological effects to plant hormones. Through artificial synthesis of substances with similar physiological and biological effects to plant hormones, they are used in agricultural production to effectively regulate the growth process of crops, achieve stable production and increase yield, improve quality, and enhance crop resistance to stress. Plant growth regulators have many advantages in use, such as: (1) wide range of effects and many application fields. Plant growth regulators can be applied to almost all higher and lower plants in the planting industry, and control the growth and development of plants by regulating the photosynthesis, respiration, absorption and movement of substances, signal transduction, stomatal opening and closing, osmotic regulation, transpiration and other physiological processes of plants, improve the interaction between plants and the environment, enhance the stress resistance of crops, increase crop yields, improve the quality of agricultural products, and make the expression of crop agronomic traits develop in the direction required by people. (2) Small dosage, fast speed, high efficiency, and less residual toxicity. (3) Can dual-regulate the external traits and internal physiological processes of plants. (4) Highly targeted and professional.

[0004] Purine itself does not exist in nature, but its derivatives are widely distributed in nature. Purine, together with specific pyrimidine bases, is a component of DNA and RNA and is very important in the process of life activities. In recent years, purine derivatives have become an important class of drugs due to their unique physiological and pharmacological properties. Pharmaceutical chemists have studied and developed a large number of nucleoside analogs with anti-tumor and anti-viral (especially anti-AIDS) effects, which has led to the rapid development of purine chemistry. In addition to the 9-position access to the sugar group of the purine ring to synthesize nucleoside drugs, there are three positions in the purine mother ring that can be used as molecular modifications. After introducing certain substituents at these positions, the resulting purine derivatives have important biomedical activities such as antiviral, anti-cancer, and anti-parasitic activities. For example, a class of purine derivatives represented by "Reversine" with R-NH-substituted derivatives at the 2 and 6 positions show very good biological activity. Therefore, the structural modification of the purine ring and the study of drug activity have always been the focus of people's research on purine compounds.

[0005] In summary, purine derivatives show high biological activity, providing a reference for the creation of new and efficient antibacterial and fungicides. Based on the previous work, the present invention designs and synthesizes a series of purine plant growth regulator compounds, and compound these compounds with D-limonene to form a compound composition. While testing its antibacterial activity, it will also study its plant growth regulator activity, hoping to screen out highly active antibacterial plant growth regulators. Summary of the invention

[0006] The present invention aims to provide a method for preparing a purine plant growth regulator 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 purine derivative compound, the derivative has the general formula of the following formula (I):

[0009]

[0010] Wherein: R1 is n-butyl, isobutyl, cyclopropylmethyl, R2 is phenyl, 4-methylphenyl, 4-methoxyphenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-nitrophenyl, 2-furyl, 2-thienyl, 3-pyridyl or a disubstituted group of any combination of the above substituents.

[0011] Preferably, R1 is n-butyl, isobutyl, cyclopropylmethyl, R2 is phenyl, 4-methylphenyl, 4-methoxyphenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-nitrophenyl, 2-furyl, 2-thienyl, 3-pyridyl or a disubstituted group of any combination of the above substituents.

[0012] A purine derivative compound, the specific compound is as follows:

[0013] Compound A1: (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide;

[0014] Compound A2: (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-methylbenzohydrazide;

[0015] Compound A3: (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-methoxybenzohydrazide;

[0016] Compound A4: (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-fluorobenzohydrazide;

[0017] Compound A5: (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-chlorobenzohydrazide;

[0018] Compound A6: (E)-4-bromo-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide;

[0019] Compound A7: (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-nitrobenzohydrazide;

[0020] Compound A8: (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)furan-2-carboxylic acid hydrazide;

[0021] Compound A9: (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)thiophene-2-carboxylic acid hydrazide;

[0022] Compound A10: (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)nicotinic acid hydrazide;

[0023] Compound A11: (E)-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide;

[0024] Compound A12: (E)-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-methylbenzohydrazide;

[0025] Compound A13: (E)-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-methoxybenzohydrazide;

[0026] Compound A14: (E)-4-fluoro-N′-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide;

[0027] Compound A15: (E)-4-chloro-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide;

[0028] Compound A16: (E)-4-bromo-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide;

[0029] Compound A17: (E)-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-nitrobenzohydrazide;

[0030] Compound A18: (E)-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)furan-2-carboxylic acid hydrazide;

[0031] Compound A19: (E)-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)thiophene-2-methylhydrazine;

[0032] Compound A20: (E)-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)nicotinic acid hydrazide;

[0033] Compound A21: (E)-N′-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide.

[0034] Compound A22: (E)-N′-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-methylbenzohydrazide.

[0035] Compound A23: (E)-N′-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-methoxybenzohydrazide.

[0036] Compound A24: (E)-N′-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-fluorobenzohydrazide.

[0037] Compound A25: (E)-4-chloro-N′-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide.

[0038] Compound A26: (E)-4-bromo-N′-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide.

[0039] Compound A27: ((E)-N′-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-nitrobenzohydrazide.

[0040] Compound A28: (E)-N′-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)furan-2-carbohydrazide.

[0041] Compound A29: ((E)-N′-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)thiophene-2-methylhydrazine.

[0042] Compound A30: (E)-N′-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)nicotinic acid hydrazide.

[0043] A method for preparing a purine derivative comprises the following steps:

[0044] (1) 6-chloro-9(H)-purine: bromoalkane: potassium carbonate = 1:1-1.2:2, 15 mL of DMF, react at room temperature for 5 h, and after the reaction is complete, pour the reaction system into saturated brine, extract with dichloromethane, collect the organic layer, and separate and purify by column chromatography to obtain the target compound 1;

[0045]

[0046] (2) Vanillin: various substituted hydrazides = 1:1-1.2 were added, and 15 mL of ethanol was refluxed for 2 h. After the reaction was completed, the reaction system solvent was concentrated under reduced pressure, the crude product was dissolved and extracted with dichloromethane, the organic layer was collected, and the target compound 2 was separated and purified by column chromatography;

[0047]

[0048] (3) Add intermediate compound 1: compound 2: potassium carbonate = 1:1:1.8, 20 mL of acetonitrile, and reflux for 7 hours. After the reaction is completed, pour the reaction system into saturated brine and extract with ethyl acetate. The organic layer is washed with saturated NaCl solution, dried over anhydrous Na2SO4, concentrated by vacuum distillation, and separated and purified by column chromatography (petroleum ether: ethyl acetate = 1:2 to 1:1) to obtain the target compound.

[0049]

[0050] The derivative is used in preparing medicines and medicaments for preventing and treating kiwi fruit bacterial canker, rice bacterial leaf streak pathogen and citrus canker pathogen.

[0051] A compound pesticide composition, characterized in that the composition comprises the purine derivative according to claim 1 and a D-limonene soluble concentrate. The mass ratio of the derivative to D-limonene is 2:1. The derivative is (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-nitrobenzoylhydrazide.

[0052] The composition is used in the preparation of a drug for preventing and treating plant bacterial diseases. The plant bacterial disease is rice streak disease.

[0053] A novel plant growth regulator is a compound (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-nitrobenzoylhydrazide, which is a compound of the following formula (II):

[0054]

[0055] Beneficial effects of the present invention: From the results of biological activity assays, it can be seen that the acylhydrazone compounds containing purine moieties have moderate to excellent inhibitory activity against rice bacterial leaf streak fungus and citrus canker fungus, among which compound A7 has the best activity, with inhibitory activities against rice bacterial leaf streak fungus and citrus canker fungus of 77.6% and 54.8%, respectively, which are higher than the control agents thiophanate-copper and thiazole zinc.

[0056] In addition, a composite composition was prepared by wettable powder of target compound A7 and D-limonene. From the biological activity test results in Table 2, it can be seen that the anti-rice bacterial leaf streak fungus and anti-citrus canker fungus activities of the composite composition are improved compared with the compounds before compounding. The inhibitory activities of composition 3 (A7: D-limonene soluble concentrate = 2:1) on rice bacterial leaf streak fungus and citrus canker fungus reached 87.6% and 75.4%, respectively. Therefore, the composite composition of A7 and D-limonene has a synergistic effect on rice bacterial leaf streak fungus and citrus canker.

[0057] Finally, the plant growth regulating activity of the compounds of the present application: the target compounds have certain plant growth regulating activity, among which compound A5 has good auxin activity, but the activity is biased compared with that of kinetin. Within a certain range, the activity of high concentration is better than that of low concentration. The activity of compound A4 and compound A5 is slightly higher than that of indoleacetic acid, which provides a basis for further research. DETAILED DESCRIPTION

[0058] Overall embodiment

[0059] 6-Chloro-9(H)-purine reacts with different bromoalkanes in DMF to generate intermediate I. Secondly, under ethanol reflux conditions, vanillin reacts with various substituted hydrazides to generate key intermediate II by Schiff base reaction. Using potassium carbonate (1.8mmol) as an acid binding agent, intermediate I (1.0mmol) and intermediate II (1.0mmol) are heated to reflux in acetonitrile solution for 7h, and the reaction progress is monitored by thin layer chromatography (TLC). After the reaction is completed, the reaction mixture is poured into water and extracted three times with ethyl acetate. The organic layer is washed with saturated NaCl solution, dried over anhydrous Na2SO4, and concentrated by vacuum distillation to obtain a crude product. Finally, the target compounds A1-A30 are purified by silica gel column chromatography using petroleum ether / ethyl acetate (1:2, v / v) as eluent.

[0060]

[0061] Specifically as follows: A method for preparing a purine derivative comprises the following steps:

[0062] (1) 6-chloro-9(H)-purine: bromoalkane: potassium carbonate = 1:1-1.2:2, 15 mL of DMF, react at room temperature for 5 h, and after the reaction is complete, pour the reaction system into saturated brine, extract with dichloromethane, collect the organic layer, and separate and purify by column chromatography to obtain the target compound 1;

[0063]

[0064] (2) Vanillin: various substituted hydrazides = 1:1-1.2 were added, and 15 mL of ethanol was refluxed for 2 h. After the reaction was completed, the reaction system solvent was concentrated under reduced pressure, the crude product was dissolved and extracted with dichloromethane, the organic layer was collected, and the target compound 2 was separated and purified by column chromatography;

[0065]

[0066] (3) Add intermediate compound 1: compound 2: potassium carbonate = 1:1:1.8, 20 mL of acetonitrile, and reflux for 7 hours. After the reaction is completed, pour the reaction system into saturated brine and extract with ethyl acetate. The organic layer is washed with saturated NaCl solution, dried over anhydrous Na2SO4, concentrated by vacuum distillation, and separated and purified by column chromatography (petroleum ether: ethyl acetate = 1:2 to 1:1) to obtain the target compound.

[0067]

[0068] Spectral data of the target compound prepared as in the general example

[0069]

[0070] (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide(A1)white solid, mp 191–192℃, yield: 61.0%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.92(s,1H),8.54(s,1H),8.52(s,1H),8.42(s,1H),7.95–7.93(m,2H),7.63–7.52(m,4H ),7.37(s,2H),4.28(t,J=7.1Hz,2H),3.77(s,3H),1.85(p,J=7.3Hz,2H),1.28(h,J=7.4Hz,2H),0.91(t,J=7.4Hz,3H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:163.2,159.1,153.2,151.7,151.2,147.3,144.9,142.4,133.4,133.2, 131.8,128.5,127.7,123.5,120.8,120.3,110.1,55.8,43.2,31.3,19.3,13.4.HRMS(ESI)m / z:[M+Na] +calcd forC 24 H 24 N6O3Na:467.1802,found:467.1789.

[0071]

[0072] (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-methylbenzohydraz ide(A2)

[0073] white solid,mp 181–182℃,yield:64.0%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.85(s,1H),8.54(s,1H),8.51(s,1H),8.42(s,1H),7.85(d,J=7.8Hz,2H),7.53(s,1H),7.36–7.33(m,4H),4.28(t,J=7.1Hz,2H),3.76(s,3H),2.39(s,3H),1.85(p,J=7.2Hz,2H),1.28(h,J=7.4Hz,2H),0.91(t,J=7.4Hz,3H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:163.0,159.1,153.2,151.7,151.2,147.1,144.9,142.4,141.9,133.2,130.5,129.0,127.7,123.5,120.8,120.3,110.1,55.8,43.2,31.3,21.1,19.3,13.4.HRMS(ESI)m / z:[M+Na] + calcd for C 25 H 26 N6O3Na:481.1959,found:481.1957.

[0074]

[0075] (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-methoxybenzohydrazide(A3)

[0076] white solid,mp 207–208℃,yield:66.4%.1 H NMR(400MHz,DMSO-d6,ppm)δ:11.80(s,1H),8.54(s,1H),8.50(s,1H),8.42(s,1H),7.95–7.92(m,2H),7.52(s,1H),7.35(s,2H),7.09–7.05(m,2H),4.28(t,J=7.1Hz,2H),3.84(s,3H),3.76(s,3H),1.85(p,J=7.3Hz,2H),1.28(h,J=7.4Hz,2H),0.91(t,J=7.4Hz,3H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:162.0,159.1,153.2,151.7,151.2,146.7,144.9,142.3,133.3,131.0,129.6,125.4,123.5,120.7,120.3,113.7,110.1,55.8,55.4,43.2,31.3,19.3,13.4.HRMS(ESI)m / z:[M+Na] + calcd for C 25 H 26 N6O4Na:497.1908,found:497.1898.

[0077]

[0078] (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-fluorobenzohydrazide(A4)

[0079] white solid,mp 192–193℃,yield:68.9%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.94(s,1H),8.54(s,1H),8.50(s,1H),8.42(s,1H),8.02(dd,J=8.7,5.6Hz,2H),7.54(s,1H),7.41–7.37(m,4H),4.28(t,J=7.1Hz,2H),3.76(s,3H),1.85(p,J=7.3Hz,2H),1.28(h,J=7.4Hz,2H),0.91(t,J=7.4Hz,3H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:164.2(d,1 J C-F =248.0Hz),162.1,159.1,153.2,151.7,151.1,147.4,144.9,142.4,133.1,130.4(d, 3 J C-F =9.0Hz),129.9(d, 4 J C-F =3.0Hz),123.5,120.8,120.3,115.5(d, 2 J C-F =22.0Hz),110.2,55.8,43.2,31.3,19.3,13.4.HRMS(ESI)m / z:[M+Na] + calcd for C 24 H 23 FN6O3Na:485.1708,found:485.1707.

[0080]

[0081] (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-chlorobenzohydrazide(A5)

[0082] white solid,mp 219–220℃,yield:65.8%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:12.03(s,1H),8.54(s,1H),8.53(s,1H),8.42(s,1H),7.97(dd,J=8.9,2.2Hz,2H),7.62(d,J=8.5Hz,2H),7.53(s,1H),7.39–7.35(m,2H),4.28(t,J=7.1Hz,2H),3.76(s,3H),1.85(p,J=7.3Hz,2H),1.28(h,J=7.4Hz,2H),0.90(t,J=7.4Hz,3H); 13C NMR(100MHz,DMSO-d6,ppm)δ:162.1,159.1,153.2,151.7,151.1,147.6,144.9,142.5,136.6,133.0,132.1,129.6,128.6,123.5,120.8,120.3,110.2,55.8,43.2,31.3,19.3,13.4.HRMS(ESI)m / z:[M+Na] + calcd for C 24 H 23 ClN6O3Na:501.1412,found:501.1405.

[0083]

[0084] (E)-4-bromo-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide(A6)

[0085] white solid,mp 179–180℃,yield:65.3%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:12.01(s,1H),8.54(s,1H),8.52(s,1H),8.42(s,1H),7.90(d,J=8.3Hz,2H),7.78–7.75(m,2H),7.53(s,1H),7.37(s,2H),4.28(t,J=7.1Hz,2H),3.76(s,3H),1.85(p,J=7.3Hz,2H),1.28(dq,J=14.5,7.4Hz,2H),0.91(t,J=7.4Hz,3H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:162.2,159.1,153.2,151.7,151.1,147.7,144.9,142.5,133.0,132.5,131.5,129.7,125.5,123.5,120.8,120.3,110.2,55.8,43.2,31.2,19.2,13.3.HRMS(ESI)m / z:[M+Na] + calcd for C 24 H 23 BrN6O3Na:545.0907,found:545.0903.

[0086]

[0087] (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-nitrobenzohydrazide(A7)

[0088] yellow solid,mp 220–221℃,yield:67.1%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:12.30(s,1H),8.57(s,1H),8.55(s,1H),8.42–8.37(m,3H),8.20(d,J=8.8Hz,2H),7.55(s,1H),7.41–7.36(m,2H),4.28(t,J=7.1Hz,2H),3.77(s,3H),1.85(p,J=7.4Hz,2H),1.28(h,J=7.4Hz,2H),0.91(t,J=7.4Hz,3H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:161.5,159.0,153.2,151.7,151.1,149.3,148.5,144.9,142.6,139.1,132.9,129.2,123.6,123.5,120.9,120.3,110.3,55.8,43.2,31.2,19.2,13.3.HRMS(ESI)m / z:[M+Na] + calcd forC 24 H 23 N7O5Na:512.1653,found:512.1647.

[0089]

[0090] (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)furan-2-carbohydrazide(A8)

[0091] white solid,mp 206–207℃,yield:62.8%. 1H NMR(400MHz,DMSO-d6,ppm)δ:11.90(s,1H),8.54(s,1H),8.50(s,1H),8.41(s,1H),7.96(d,J=1.0Hz,1H),7.50(s,1H),7.37–7.32(m,3H),6.71(dd,J=3.5,1.7Hz,1H),4.28(t,J=7.1Hz,2H),3.76(s,3H),1.85(p,J=7.3Hz,2H),1.28(h,J=7.4Hz,2H),0.91(t,J=7.4Hz,3H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:159.1,154.2,153.2,151.7,151.2,147.4,146.6,145.9,144.9,142.4,133.1,123.6,120.8,120.3,115.0,112.1,110.1,55.8,43.2,31.3,19.3,13.4.HRMS(ESI)m / z:[M+Na] + calcd for C 22 H 22 N6O4Na:457.1595,found:457.1594.

[0092]

[0093] (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)thiophene-2-carbohydrazide(A9)

[0094] white solid,mp 186–188℃,yield:62.6%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.95(s,1H),8.56–7.89(m,5H),7.58(d,J=42.0Hz,1H),7.53–7.37(m,2H),7.23(s,1H),4.28(t,J=7.1Hz,2H),3.78(s,3H),1.85(p,J=7.3Hz,2H),1.28(h,J=7.4Hz,2H),0.91(t,J=7.4Hz,3H); 13C NMR(100MHz,DMSO-d6,ppm)δ:161.4,159.1,153.2,151.7,151.2,147.1,144.9,143.2,135.1,134.9,133.0,132.8,131.9,126.6,123.7,120.3,111.0,55.8,43.2,31.3,19.3,13.4.HRMS(ESI)m / z:[M+Na] + calcd for C 22 H 22 N6O3SNa:473.1366,found:473.1357.

[0095]

[0096] (E)-N'-(4-((9-butyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)nicotinohydrazide(A10)

[0097] white solid,mp 150–151℃,yield:58.6%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:12.22(s,1H),9.11(d,J=2.3Hz,1H),8.77(dd,J=4.8,1.7Hz,1H),8.56(d,J=6.5Hz,2H),8.42(s,1H),8.31(dt,J=8.0,2.0Hz,1H),7.60–7.54(m,2H),7.40–7.36(m,2H),4.28(t,J=7.1Hz,2H),3.77(s,3H),1.85(p,J=7.3Hz,2H),1.28(h,J=7.4Hz,2H),0.90(t,J=7.4Hz,3H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:161.8,159.1,153.2,152.3,151.7,151.1,148.7,148.0,144.9,142.5,135.5,133.0,129.2,123.6,123.5,120.9,120.3,110.3,55.8,43.2,31.3,19.3,13.4.HRMS(ESI)m / z:[M+Na] + calcd for C 23 H 23N7O3Na:468.1755,found:468.1751.

[0098]

[0099] (E)-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide(A11)

[0100] white solid,mp 191–192℃,yield:56.8%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.92(s,1H),8.52(d,J=3.6Hz,2H),8.42(s,1H),7.94(d,J=7.2Hz,2H),7.63–7.52(m,4H),7.37(s,2H),4.11(d,J=7.2Hz,2H),3.77(s,3H),2.25(hept,J=6.8Hz,1H),0.88(d,J=6.7Hz,6H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:163.2,159.1,153.4,151.7,151.2,147.3,145.2,142.4,133.4,133.2,131.8,128.5,127.6,123.5,120.8,120.2,110.1,56.0,50.5,28.5,19.6.HRMS(ESI)m / z:[M+Na] + calcd for C 24 H 24 N6O3Na:467.1802,found:467.1805.

[0101]

[0102] (E)-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-methylbenzohydrazide(A12)

[0103] white solid,mp 169–170℃,yield:55.6%. 1H NMR(400MHz,DMSO-d6,ppm)δ:12.02(s,1H),8.59(s,1H),8.53(s,1H),8.41(s,1H),7.91–7.88(m,2H),7.52(s,1H),7.38–7.32(m,4H),4.11(d,J=7.2Hz,2H),3.76(s,3H),2.38(s,3H),2.24(hept,J=6.8Hz,1H),0.88(d,J=6.7Hz,6H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:163.0,159.1,153.4,151.7,151.2,147.1,145.2,142.3,141.8,133.3,130.5,129.0,127.8,123.5,120.7,120.2,110.1,55.8,50.5,28.5,21.1,19.7.HRMS(ESI)m / z:[M+Na] + calcd for C 25 H 26 N6O3Na:481.1959,found:481.1964.

[0104]

[0105] (E)-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-methoxybenzohydrazide(A13)

[0106] white solid,mp 150–151℃,yield:59.6%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.79(s,1H),8.51(d,J=8.3Hz,2H),8.42(s,1H),7.95–7.92(m,2H),7.52(s,1H),7.36(s,2H),7.09–7.06(m,2H),4.11(d,J=7.2Hz,2H),3.84(s,3H),3.76(s,3H),2.25(hept,J=6.8Hz,1H),0.88(d,J=6.6Hz,6H); 13C NMR(100MHz,DMSO-d6,ppm)δ:162.0,159.1,153.4,151.7,151.2,146.7,145.2,142.3,136.8,133.3,129.6,125.4,123.5,120.7,120.2,113.7,110.1,55.8,55.4,50.5,28.5,19.6.HRMS(ESI)m / z:[M+Na] + calcd forC 25 H 26 N6O4Na:497.1908,found:497.1902.

[0107]

[0108] (E)-4-fluoro-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide(A14)

[0109] white solid,mp 204–205℃,yield:62.5%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.94(s,1H),8.51(d,J=7.9Hz,2H),8.42(s,1H),8.02(dd,J=8.6,5.6Hz,2H),7.54(s,1H),7.41–7.36(m,4H),4.11(d,J=7.2Hz,2H),3.77(s,3H),2.25(hept,J=6.8Hz,1H),0.88(d,J=6.7Hz,6H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:164.2(d, 1 J C-F =246.0Hz),162.1,159.1,153.4,151.7,151.2,147.4,145.2,142.4,133.1,130.4(d, 3 J C-F =9.0Hz),129.9(d, 4 J C-F =3.0Hz),123.5,120.8,120.2,115.5(d, 2 J C-F=21.0Hz),110.2,56.0,50.5,28.5,19.6.HRMS(ESI)m / z:[M+Na] + calcd for C 24 H 23 FN6O3Na:485.1708,found:485.1703.

[0110]

[0111] (E)-4-chloro-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide(A15)

[0112] white solid,mp 149–150℃,yield:61.0%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.99(s,1H),8.51(d,J=7.7Hz,2H),8.42(s,1H),7.97(d,J=8.2Hz,2H),7.62(d,J=8.2Hz,2H),7.54(s,1H),7.37(s,2H),4.11(d,J=7.2Hz,2H),3.77(s,3H),2.25(hept,J=6.8Hz,1H),0.88(d,J=6.7Hz,6H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:162.1,159.1,153.4,151.7,151.2,147.6,145.2,142.5,136.6,133.0,132.1,129.6,128.6,123.5,120.8,120.2,110.2,55.8,50.5,28.5,19.6.HRMS(ESI)m / z:[M+Na] + calcd for C 24 H 23 ClN6O3Na:501.1412,found:501.1408.

[0113]

[0114] (E)-4-bromo-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide(A16)

[0115] white solid,mp 158–159℃,yield:60.7%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.98(s,1H),8.51(d,J=8.6Hz,2H),8.41(s,1H),7.89(d,J=8.4Hz,2H),7.77(d,J=8.4Hz,2H),7.54(s,1H),7.37(s,2H),4.11(d,J=7.2Hz,2H),3.76(s,3H),2.25(hept,J=6.8Hz,1H),0.88(d,J=6.7Hz,6H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:162.2,159.1,153.4,151.7,151.1,147.6,145.2,142.5,133.0,132.5,131.5,129.7,125.5,123.5,120.8,120.2,110.2,55.8,50.5,28.4,19.6.HRMS(ESI)m / z:[M+Na] + calcd for C 24 H 23 BrN6O3Na:545.0907,found:545.0901.

[0116]

[0117] (E)-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-nitrobenzohydrazide(A17)

[0118] yellow solid,mp 233–234℃,yield:56.9%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:12.28(s,1H),8.55(d,J=13.2Hz,2H),8.42–8.38(m,3H),8.19(d,J=8.6Hz,2H),7.55(s,1H),7.39(s,2H),4.11(d,J=7.1Hz,2H),3.77(s,3H),2.25(hept,J=6.8Hz,1H),0.88(d,J=6.7Hz,6H); 13C NMR(100MHz,DMSO-d6,ppm)δ:161.6,159.1,153.4,151.7,151.2,149.3,148.4,145.2,142.6,139.1,132.9,129.2,123.7,123.6,121.0,120.2,110.3,55.8,50.5,28.5,19.6.HRMS(ESI)m / z:[M+Na] + calcd for C 24 H 23 N7O5Na:512.1653,found:512.1651.

[0119]

[0120] (E)-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)furan-2-carbohydrazide(A18)

[0121] white solid,mp 211–213℃,yield:59.0%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.92(s,1H),8.51(d,J=6.3Hz,2H),8.41(s,1H),7.96(s,1H),7.51(s,1H),7.38–7.33(m,3H),6.71(dd,J=3.5,1.8Hz,1H),4.11(d,J=7.2Hz,2H),3.76(s,3H),2.24(hept,J=6.8Hz,1H),0.88(d,J=6.7Hz,6H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:159.1,154.3,153.4,151.7,151.2,147.4,146.6,145.9,145.2,142.4,133.1,123.5,120.8,120.2,115.0,112.1,110.1,55.8,50.5,28.5,19.6.HRMS(ESI)m / z:[M+Na] + calcd for C 22 H 22 N6O4Na:457.1595,found:457.1584.

[0122]

[0123] (E)-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)thiophene-2-carbohydrazide(A19)

[0124] white solid,mp 181–182℃,yield:58.9%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.95(s,1H),8.52(s,1H),8.41(s,1H),8.16–7.89(m,3H),7.58(d,J=41.3Hz,1H),7.44–7.37(m,2H),7.23(t,J=4.3Hz,1H),4.11(d,J=7.2Hz,2H),3.78(s,3H),2.25(hept,J=6.8Hz,1H),0.88(d,J=6.8Hz,6H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:159.1,153.4,151.7,151.2,147.1,145.2,143.2,142.2,135.1,133.0,132.0,129.0,126.6,123.6,120.8,120.2,111.1,55.8,50.5,28.5,19.6.HRMS(ESI)m / z:[M+Na] + calcd for C 22 H 22 N6O3SNa:473.1366,found:473.1356.

[0125]

[0126] (E)-N'-(4-((9-isobutyl-9H-purin-6-yl)oxy)-2-methoxybenzylidene)nicotinohydrazide(A20)

[0127] white solid,mp 187–188℃,yield:54.6.0%. 1H NMR(400MHz,DMSO-d6,ppm)δ:12.09(s,1H),9.09(d,J=2.2Hz,1H),8.78(dd,J=4.8,1.7Hz,1H),8.51(d,J=9.8Hz,2H),8.42(s,1H),8.28(dt,J=8.1,2.0Hz,1H),7.60–7.55(m,2H),7.39(s,2H),4.11(d,J=7.1Hz,2H),3.77(s,3H),2.25(hept,J=6.8Hz,1H),0.88(d,J=6.7Hz,6H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:161.8,159.1,153.4,152.3,151.7,151.2,148.6,147.9,145.2,142.5,135.5,132.9,129.2,123.6,123.6,120.9,120.2,110.3,55.8,50.5,28.5,19.6.HRMS(ESI)m / z:[M+Na] + calcd for C 23 H 23 N7O3Na:468.1755,found:468.1745.

[0128]

[0129] (E)-N'-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide(A21)

[0130] white solid,mp 189–190℃,yield:61.8%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.93(s,1H),8.59(s,1H),8.52(s,1H),8.43(s,1H),7.95–7.93(m,2H),7.63–7.52(m,4H),7.37(s,2H),4.15(d,J=7.3Hz,2H),3.77(s,3H),1.41–1.31(m,1H),0.58–0.52(m,2H),0.52–0.46(m,2H); 13C NMR(100MHz,DMSO-d6,ppm)δ:163.2,159.1,153.1,151.7,151.2,147.3,144.6,142.4,133.4,133.2,131.8,128.5,127.6,123.5,120.8,120.2,110.1,55.8,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd for C 24 H 22 N6O3Na:465.1646,found:465.1644.

[0131]

[0132] (E)-N'-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-methylbenzohydrazide(A22)

[0133] white solid,mp 203–204℃,yield:62.9%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.85(s,1H),8.58(s,1H),8.51(s,1H),8.43(s,1H),7.85(d,J=7.9Hz,2H),7.53(s,1H),7.36–7.33(m,4H),4.15(d,J=7.3Hz,2H),3.77(s,3H),2.39(s,3H),1.41–1.31(m,1H),0.58–0.50(m,2H),0.52–0.46(m,2H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:163.0,159.1,153.1,151.7,151.2,147.0,144.6,142.3,141.8,133.2,130.5,129.0,127.6,123.5,120.7,120.2,110.1,55.8,47.9,21.0,11.2,3.8.HRMS(ESI)m / z:[M+Na] + calcd forC 25 H 24 N6O3Na:479.1802,found:479.1798.

[0134]

[0135] (E)-N'-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-methoxybenzohydrazide(A23)

[0136] white solid,mp 181–183℃,yield:60.3%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.98(s,1H),8.59(s,2H),8.42(s,1H),7.98(d,J=7.9Hz,2H),7.51(s,1H),7.35(s,2H),7.08–7.05(m,2H),4.15(d,J=7.3Hz,2H),3.84(s,3H),3.76(s,3H),1.41–1.31(m,1H),0.57–0.52(m,2H),0.51–0.46(m,2H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:162.0,159.1,153.1,151.7,151.2,146.7,144.7,142.2,138.3,133.3,129.7,125.4,123.5,120.6,120.2,113.7,110.0,55.8,55.4,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd forC 25 H 24 N6O4Na:495.1751,found:495.1744.

[0137]

[0138] (E)-N'-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-fluorobenzohydrazide(A24)

[0139] white solid,mp 187–189℃,yield:66.0%. 1H NMR(400MHz,DMSO-d6,ppm)δ:12.10(s,1H),8.59(d,J=3.6Hz,2H),8.42(s,1H),8.06(dd,J=8.7,5.5Hz,2H),7.53(s,1H),7.40–7.35(m,4H),4.15(d,J=7.3Hz,2H),3.77(s,3H),1.40–1.32(m,1H),0.57–0.52(m,2H),0.50–0.46(m,2H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:164.2(d, 1 J C-F =247.0Hz),162.1,159.1,153.1,151.7,151.2,147.4,144.7,142.4,133.1,130.4(d, 3 J C-F =9.0Hz),129.8,123.5,120.8,120.2,115.4(d, 2 J C-F =22.0Hz),110.1,55.8,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd for C 24 H 21 FN6O3Na:483.1551,found:483.1550.

[0140]

[0141] (E)-4-chloro-N'-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide(A25)

[0142] white solid,mp 170–171℃,yield:62.0%. 1H NMR(400MHz,DMSO-d6,ppm)δ:12.01(s,1H),8.58(s,1H),8.51(s,1H),8.42(s,1H),7.97(d,J=8.2Hz,2H),7.62(d,J=8.2Hz,2H),7.54(s,1H),7.37(s,2H),4.15(d,J=7.3Hz,2H),3.77(s,3H),1.41–1.31(m,1H),0.57–0.53(m,2H),0.50–0.46(m,2H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:162.2,159.1,153.1,151.7,151.2,147.6,144.6,142.4,136.5,133.1,132.2,129.6,128.5,123.5,120.8,120.2,110.2,55.8,47.9,11.2,3.8.HRMS(ESI)m / z:[M+Na] + calcd forC 24 H 21 ClN6O3Na:499.1256,found:499.1251.

[0143]

[0144] (E)-4-bromo-N'-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)benzohydrazide(A26)

[0145] white solid,mp 225–226℃,yield:61.1%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.97(s,1H),8.58(s,1H),8.51(s,1H),8.42(s,1H),7.89(d,J=8.2Hz,2H),7.76(d,J=8.3Hz,2H),7.54(s,1H),7.37(s,2H),4.15(d,J=7.3Hz,2H),3.77(s,3H),1.41–1.31(m,1H),0.57–0.52(m,2H),0.52–0.46(m,2H); 13C NMR(100MHz,DMSO-d6,ppm)δ:162.2,159.1,153.1,151.7,151.2,147.7,144.6,142.5,133.0,132.5,131.5,129.7,125.5,123.5,120.8,120.2,110.2,55.8,47.9,11.2,3.8.HRMS(ESI)m / z:[M+Na] + calcd forC 24 H 21 BrN6O3Na:543.0751,found:543.0739.

[0146]

[0147] (E)-N'-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)-4-nitrobenzohydrazide(A27)

[0148] yellow solid,mp 229–230℃,yield:63.9%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:12.22(s,1H),8.59(s,1H),8.53(s,1H),8.43(s,1H),8.39(d,J=8.9Hz,2H),8.18(d,J=8.8Hz,2H),7.56(d,J=1.6Hz,1H),7.42–7.37(m,2H),4.15(d,J=7.3Hz,2H),3.78(s,3H),1.41–1.31(m,1H),0.58–0.52(m,2H),0.52–0.46(m,2H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:161.6,159.1,153.1,151.7,151.2,149.3,148.4,144.7,142.6,139.1,132.9,129.2,123.7,123.6,121.0,120.2,110.3,55.8,47.9,11.3,3.9.HRMS(ESI)m / z:[M+Na] + calcdfor C 24 H 21 N7O5Na:510.1496,found:510.1493.

[0149]

[0150] (E)-N'-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)furan-2-carbohydrazide(A28)

[0151] white solid,mp 214–216℃,yield:62.6%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:11.90(s,1H),8.58(s,1H),8.50(s,1H),8.42(s,1H),7.96(d,J=1.8Hz,1H),7.51(s,1H),7.34(d,J=13.6Hz,3H),6.71(dd,J=3.6,1.8Hz,1H),4.14(d,J=7.3Hz,2H),3.77(s,3H),1.41–1.31(m,1H),0.57–0.52(m,2H),0.52–0.46(m,2H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:159.1,154.2,153.1,151.7,151.2,147.4,146.6,145.9,144.7,142.4,133.1,123.6,120.8,120.2,115.0,112.1,110.1,55.8,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcdfor C 22 H 20 N6O4Na:455.1438,found:455.1440.

[0152]

[0153] (E)-N'-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)thiophene-2-carbohydrazide(A29)

[0154] white solid,mp 221–222℃,yield:64.1%. 1H NMR(400MHz,DMSO-d6,ppm)δ:11.96(s,1H),8.59(s,1H),8.42(s,1H),8.16–7.88(m,3H),7.58(d,J=43.4Hz,1H),7.44–7.37(m,2H),7.23(t,J=4.4Hz,1H),4.15(d,J=7.3Hz,2H),3.79(s,3H),1.41–1.31(m,1H),0.57–0.52(m,2H),0.52–0.46(m,2H); 13 C NMR(100MHz,DMSO-d6,ppm)δ:161.4,159.1,153.1,151.7,151.2,147.1,144.7,143.2,135.1,133.0,132.8,129.1,128.2,126.6,123.6,120.2,111.0,55.8,47.9,11.3,3.8.HRMS(ESI)m / z:[M+Na] + calcd forC 22 H 20 N6O3SNa:471.1210,found:471.1207.

[0155]

[0156] (E)-N'-(4-((9-(cyclopropylmethyl)-9H-purin-6-yl)oxy)-2-methoxybenzylidene)nicotinohydrazide(A30)

[0157] white solid,mp 202–203℃,yield:60.8%. 1 H NMR(400MHz,DMSO-d6,ppm)δ:12.09(s,1H),9.09(s,1H),8.78(dd,J=4.9,1.6Hz,1H),8.59(s,1H),8.50(s,1H),8.43(s,1H),8.28(d,J=8.0Hz,1H),7.60–7.56(m,2H),7.41–7.36(m,2H),4.15(d,J=7.3Hz,2H),3.77(s,3H),1.41–1.31(m,1H),0.58–0.52(m,2H),0.52–0.46(m,2H); 13C NMR(100MHz,DMSO-d6,ppm)δ:161.8,159.1,153.1,152.3,151.7,151.2,148.6,147.9,144.7,142.5, 135.5,132.9,129.2,123.6,123.6,120.9,120.2,110.3,55.8,47.9,11.3,3.9.HRMS(ESI)m / z:[M+Na] + Calculate for C 23 H 21 N7O3Na:466.1598,found:466.1592.

[0158] 3. Antimicrobial activity data of target compounds

[0159] The inhibitory activity of the target compounds against kiwifruit bacterial canker, rice bacterial leaf streak and citrus canker was determined by turbidimetric method. Pure dimethyl sulfoxide (DMSO) was used as negative control, and agricultural fungicides zinc thiazole and thiophanate-copper were used as positive controls. The compounds were tested at 100 and 50 μg / mL concentrations. Preparation of NB medium: 15 g glucose, 7.5 g peptone, 1.5 g yeast powder and 4.5 g beef extract were dissolved in 1500 mL secondary water, pH was adjusted to 7.0-7.2, and sterilized in an autoclave at 121°C for 30 min. 40 μL NB medium (containing bacteria Psa, Xoc or Xac) was pipetted and mixed with a mixture containing 4 mL NB medium and 1 mL 0.1% tween-20 (containing the compound). The above solution was incubated at 28°C / 180rpm for 1-3 days, and the bacterial growth was monitored. When the OD value of the blank control is between 0.6 and 0.8, the OD value of the compound is measured at a wavelength of 595 nm (200 μL of the solution is added to a 96-well plate enzyme counter and the absorbance is measured), and the inhibition rate is calculated according to the following formula:

[0160] I=(C tur -T tur ) / C tur ×100%

[0161] Among them, C tur The blank control group, T tur The drug-treated group, I tur is the inhibition rate.

[0162] Table 1. In vitro antibacterial activity of compounds A1-A30 against Xanthomonas oryzae, Xanthomonas citri var. citri and Xanthomonas avium

[0163]

[0164]

[0165] From the biological activity test results in Table 1, it can be seen that the acylhydrazone compounds containing purine moieties have moderate to excellent inhibitory activity against rice bacterial leaf streak fungus and citrus canker fungus, among which compound A7 has the best activity, with inhibitory activities against rice bacterial leaf streak fungus and citrus canker fungus of 77.6% and 54.8%, respectively, which are higher than the control agents thiophanate-copper and thiazole zinc.

[0166] 4. Preparation of target compound A7 composition

[0167] The wettable powder of target compound A7 and D-limonene is used to prepare a composite composition. In each composition, the ratio of target compound A7 and D-limonene is calculated according to the mass ratio. The following preparations of target compound A7 and D-limonene are prepared as needed.

[0168] Composition 1: A7: D-limonene soluble solution = 1:1

[0169] Composition 2: A7: D-limonene soluble solution = 1:2

[0170] Composition 3: A7: D-limonene soluble solution = 2:1

[0171] Anti-rice bacterial leaf streak disease and citrus canker disease activity of the composition of target compound A7

[0172] The antimicrobial activity of the composition of target compound A7 and D-limonene against Psoralea oryzae oryzae and Psoralea citri citri were tested according to the turbidity method (the antimicrobial activity test method mentioned previously).

[0173] Table 2 Inhibitory activity of drugs against rice bacterial leaf streak pathogen and citrus canker

[0174]

[0175]

[0176] The in vitro growth rate method was used to test the activity of the composition against rice bacterial leaf streak and citrus canker 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 and citrus canker was improved compared with the compounds before compounding. The inhibitory activity of composition 3 (A7: D-limonene soluble solution = 2:1) against rice bacterial leaf streak and citrus canker reached 87.6% and 75.4%, respectively. Therefore, the composite composition of A7 and D-limonene has a synergistic effect on rice bacterial leaf streak and citrus canker.

[0177] 5. Determination of plant growth regulating activity of some target compounds

[0178] (1) Test method

[0179] 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.

[0180] (2) Wheat bud sheath cutting test method:

[0181] 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 1 hour. Then, measure 8mL of each sample solution and place them in a culture dish with a diameter of 9cm. Place two filter paper pieces in each culture dish, then evenly place the cut 10 bud sheath segments in each treated culture dish, and then place them 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:

[0182] Effect = (treatment - blank) / blank × 100%

[0183] 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:

[0184]

[0185] (3) Radish cotyledon expansion test method:

[0186] 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.

[0187] (3) Biological test results

[0188] Table 3 Plant growth regulator activity of some target compounds

[0189]

[0190]

[0191] 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 A5 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. The activity of compound A4 and compound A5 is slightly higher than that of indoleacetic acid, which can be further studied.

Claims

1. A purine plant growth regulator, characterized in that: The structural formula of the purine plant growth regulator is as follows: , wherein R1 is C3-C4 alkyl and cycloalkyl, and R2 is phenyl, substituted phenyl, or heterocyclic.

2. A purine plant growth regulator according to claim 1, characterized in that: The R1 is C4 alkyl, isopropyl and cyclopropylmethyl, R2 is phenyl, substituted phenyl, heterocyclic group, the substituent of the substituted phenyl is methyl, halogen or nitro, and the heterocyclic group is thiophene, furan or pyridine.

3. The method for preparing a purine plant growth regulator according to claim 1, characterized in that: The reaction formula is as follows:

4. The method for preparing a purine plant growth regulator according to claim 3, characterized in that: The method comprises the following steps: firstly, using potassium carbonate as an acid-binding agent, 6-chloro-9(H)-purine reacts with different bromoalkanes in DMF to generate an intermediate I; secondly, under ethanol reflux conditions, vanillin reacts with various substituted hydrazides to generate a key intermediate II by Schiff base reaction; finally, using potassium carbonate as an acid-binding agent, the intermediate I is combined with different Schiff bases (II) in acetonitrile to obtain a target compound.

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

6. A compound pesticide composition, characterized in that: The composition comprises the purine plant growth regulator according to claim 1 and D-limonene.

7. The composition according to claim 6, characterized in that: The mass ratio of the derivative to D-limonene is 1:2-2:

1.

8. Use of the composition according to claim 6 in the preparation of a medicament for preventing and controlling oxysporum leaf streak of rice and citrus canker.

9. The composition according to claim 6, characterized in that: The derivative is A7.

10. Application of a purine plant growth regulator as claimed in claim 1 or 2 in the preparation of a plant growth regulator.

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