Pyrazolopyrimidone derivative containing aniline unit as well as preparation method and application of pyrazolopyrimidone derivative

Abstract

The invention belongs to the field of pesticides, and discloses a compound with a structure shown as a formula I. A preparation method of the compound comprises the following steps: sequentially carrying out hydrolysis, amidation and cyclization on 5-amino-1-methylpyrazole-4-ethyl formate to obtain a pyrazolopyrimidone derivative containing an aniline unit; the pyrazolopyrimidone derivative containing the aniline unit is a novel compound, has an inhibiting effect on gibberellic disease bacteria, rhizoctonia solani and botrytis cinerea, and can be used for preventing and treating plant diseases caused by fungi. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of pesticides, and particularly relates to a pyrazolopyrimidone derivative containing an aniline unit, and a preparation method and application thereof. Background Art

[0002] Plant fungal diseases are an important factor affecting crop yield and quality, and have caused huge agricultural production losses worldwide. Therefore, timely and effective prevention and control of crop fungal diseases is of vital importance to ensure the smooth progress of agricultural production. At present, the means of preventing and controlling plant fungal diseases are still mainly chemical control. However, the long-term use of fungicides such as boscalid, azoxystrobin, and carbendazim has caused serious drug resistance in plant pathogenic fungi. Therefore, there is an urgent need to develop new green fungicides with novel molecular structures and unique mechanisms of action. Summary of the invention

[0003] Purpose of the invention: The first purpose of the present invention is to provide a pyrazolopyrimidone derivative containing an aniline unit which has an inhibitory effect on plant pathogenic fungal diseases; the second purpose of the present invention is to provide a method for preparing the pyrazolopyrimidone derivative containing an aniline unit; the third purpose of the present invention is to provide the application of the pyrazolopyrimidone derivative containing an aniline unit in preventing and controlling plant pathogenic fungal diseases.

[0004] Technical solution: The present invention discloses a pyrazolopyrimidone derivative containing an aniline unit, the structural formula of which is shown in I:

[0005]

[0006] Among them, R 1 is selected from hydrogen, 2-methyl, 3-methyl, 4-methyl, 2,4-dimethyl, 2-chloro, 3-chloro, 4-chloro, 2,4-dichloro, 3,4-dichloro, 3,5-dichloro, 2-fluoro, 4-fluoro, 2,4-difluoro, 2-bromo or 2-methoxy.

[0007] The preparation method of the pyrazolopyrimidone derivative containing an aniline unit of the present invention comprises the following steps: 5-amino-1-methylpyrazole-4-carboxylic acid ethyl ester is hydrolyzed with a 5% sodium hydroxide solution, and then reacted with a substituted phenylhydrazine in dichloromethane containing triethylamine and TBTU; the obtained hydrazide molecule is cyclized with triethyl orthoformate in hot ethanol to obtain the pyrazolopyrimidone derivative containing an aniline unit. The synthesis route is as follows:

[0008]

[0009] The invention discloses an application of the pyrazolopyrimidone derivative containing aniline unit in preventing and controlling plant pathogenic fungal diseases.

[0010] The plant pathogenic fungi are wheat fusarium rust, rice sheath blight and tomato gray mold.

[0011] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: (1) the pyrazolopyrimidone derivatives containing aniline units described in the present invention are a new type of compound, which has an inhibitory effect on wheat fusarium, rice sheath blight or tomato gray mold; (2) the preparation method of the pyrazolopyrimidone derivatives containing aniline units described in the present invention is simple, the raw materials are easily available, and the reaction is novel and efficient; (3) the pyrazolopyrimidone derivatives containing aniline units described in the present invention can significantly inhibit wheat fusarium, and the optimal inhibition rate at a dosage of 50 μg / mL is 91.63%, which has an application prospect for development as a potential agricultural fungicide. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 This is the hydrogen spectrum of the pyrazolopyrimidone derivative containing aniline unit prepared in Example 1;

[0013] Figure 2 This is the carbon spectrum of the pyrazolopyrimidone derivative containing aniline unit prepared in Example 1;

[0014] Figure 3 This is a high-resolution mass spectrum of the pyrazolopyrimidone derivative containing an aniline unit prepared in Example 1;

[0015] Figure 4 This is a graph showing the effect of the pyrazolopyrimidone derivative containing aniline units prepared in Example 13 on inhibiting wheat fusarium at different dosage concentrations. DETAILED DESCRIPTION

[0016] The technical solution of the present invention is further described below in conjunction with embodiments.

[0017] Example 1

[0018] The pyrazolopyrimidinone derivative I1 containing an aniline unit of the present invention, R1 = H, has a chemical name of 1-methyl-5-(anilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, an English name of 1-methyl-5-(phenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, and has the following structural formula:

[0019]

[0020] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0021]

[0022] Add 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (18 mmol) and 5% NaOH solution (10 mL) to 100 mL of ethanol, heat to 75-80°C and react for about 4-5 hours. After TLC monitoring, the raw material disappears, and the mixture is allowed to stand overnight to produce a large amount of white floccules. After filtration, the filter cake is dissolved in water, and the pH of the system is adjusted to 5-6 with 5% HCl solution, and a large amount of white solid is precipitated. Filter and dry the obtained filter cake to obtain 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (intermediate A).

[0023] Step 2: Synthesis of 5-amino-1-methyl-N′-phenyl-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B1)

[0024]

[0025] 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (7mmol), phenylhydrazine (7mmol), TBTU condensing agent (8mmol) and dichloromethane (30mL) were added to a 100mL round-bottom bottle in sequence and stirred at room temperature for about 30 minutes. Triethylamine (1mL) was then added dropwise to the above mixture and stirring continued at room temperature. After TLC monitoring of the disappearance of the reaction raw materials, the excess solvent was removed by reduced pressure distillation, and the residue was purified and separated by thin layer chromatography to obtain 5-amino-1-methyl-N′-phenyl-1H-pyrazole-4-carboxylic acid hydrazide (intermediate B1).

[0026] Step 3: Synthesis of 1-methyl-5-(anilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I1)

[0027] 5-Amino-1-methyl-N′-phenyl-1H-pyrazole-4-carboxylic acid hydrazide (2 mmol) and triethyl orthoformate (6 mmol) were dissolved in anhydrous ethanol (30 mL), and then heated to 75-80°C for about 4-5 hours. After the raw material disappeared as monitored by TLC, the mixture was allowed to stand overnight to precipitate a large amount of white solid. The filter cake was filtered and recrystallized with an appropriate amount of ethanol to obtain the target compound I1.

[0028] The target compound I1 is a white solid; yield 61%; mp 216-218°C; 1 H NMR (400MHz, DMSO-d6) δ9.28 (s, 1H), 8.50 (s, 1H), 8.15 (s, 1H), 7.19 (t, J = 7.8Hz, 2H), 6.85 (t, J = 7.3Hz, 1H), 6.64 (d, J = 7.9Hz, 2H), 3.96 (s, 3H); 13C NMR (100MHz, DMSO-d6) δ156.09, 153.34, 151.25, 148.16, 135.00, 129.57, 121.02, 113.18, 106.22, 34.63; HRMS (ESI) m / z [M+H] + Calculate for C 12 H 11 N5O: 242.1036, found: 242.1038.

[0029] Example 2

[0030] The pyrazolopyrimidinone derivative I2 containing an aniline unit of the present invention, R1 = 4-Cl, has a chemical name of 1-methyl-5-(4-chloroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, an English name of 1-methyl-5-(4-chlorophenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, and has the following structural formula:

[0031]

[0032] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0033] As in the first step of Example 1.

[0034] Step 2: Synthesis of 5-amino-1-methyl-N′-(4-chlorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B2)

[0035]

[0036] As in the second step of Example 1, the difference is that 4-chlorophenylhydrazine is used as the reactant.

[0037] Step 3: Synthesis of 1-methyl-5-(4-chloroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I2)

[0038] The same as the third step of Example 1, except that 5-amino-1-methyl-N′-(4-chlorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0039] The target compound I2 is a white solid; yield 67%; mp 242-244°C; 1H NMR (400MHz, DMSO-d6) δ9.43 (s, 1H), 8.50 (s, 1H), 8.15 (s, 1H), 7.23 (d, J = 8.4Hz, 2H), 6.68 (d, J = 8.4Hz, 2H), 3.96 (s, 3H); 13 C NMR (100MHz, DMSO-d6) δ155.97, 153.18, 151.22, 147.13, 135.05.129.35, 124.58, 114.92, 106.17, 34.63; HRMS (ESI) m / z [M+H] + Calculate for C 12 H 10 ClN5O: 276.0647, found: 276.0648.

[0040] Example 3

[0041] The pyrazolopyrimidinone derivative I3 containing an aniline unit of the present invention, R1 = 4-Me, has a chemical name of 1-methyl-5-(4-methylanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, an English name of 1-methyl-5-(4-methylphenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, and has the following structural formula:

[0042]

[0043] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0044] As in the first step of Example 1.

[0045] Step 2: Synthesis of 5-amino-1-methyl-N′-(4-methylphenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B3)

[0046]

[0047] The process is the same as the second step of Example 1, except that 4-methylphenylhydrazine is used as the reactant.

[0048] Step 3: Synthesis of 1-methyl-5-(4-methylanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I3)

[0049] The process is the same as the third step of Example 1, except that 5-amino-1-methyl-N′-(4-methylphenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0050] The target compound I3 is a white solid; yield 63%; mp 243-245°C; 1 H NMR (400MHz, DMSO-d6) δ9.13 (s, 1H), 8.48 (s, 1H), 8.13 (s, 1H), 6.99 (d, J = 7.4Hz, 2H), 6.55 (d, J = 7.5Hz, 2H), 3.96 (s, 3H), 2.18 (s, 3H); 13 C NMR (100MHz, DMSO-d6) δ156.11, 153.32, 151.25, 145.84, 134.96, 129.93, 129.83, 113.41, 106.23, 34.61, 20.59; HRMS (ESI) m / z [M+H] + Calculate for C 13 H 13 N5O: 256.1193, found: 256.1195.

[0051] Example 4

[0052] The pyrazolopyrimidinone derivative I4 containing an aniline unit of the present invention, R1 = 2-Cl, has a chemical name of 1-methyl-5-(2-chloroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, an English name of 1-methyl-5-(2-chlorophenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, and has the following structural formula:

[0053]

[0054] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0055] As in the first step of Example 1.

[0056] Step 2: Synthesis of 5-amino-1-methyl-N′-(2-chlorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B4)

[0057]

[0058] As in the second step of Example 1, the difference is that 2-chlorophenylhydrazine is used as the reactant.

[0059] Step 3: Synthesis of 1-methyl-5-(2-chloroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I4)

[0060] The same as the third step of Example 1, except that 5-amino-1-methyl-N′-(2-chlorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0061] The target compound I4 is a white solid; yield 59%; mp 237-239°C; 1 H NMR (400MHz, DMSO-d6) δ8.96 (s, 1H), 8.53 (s, 1H), 8.16 (s, 1H), 7.41 (d, J=7.8Hz, 1H ), 7.11 (t, J=7.6Hz, 1H), 6.88 (t, J=7.5Hz, 1H), 6.44 (d, J=8.0Hz, 1H), 3.97 (s, 3H); 13 C NMR (100MHz, DMSO-d6) δ155.67, 153.28, 151.18, 143.67, 135.10, 130.07, 128.51, 121.90, 118.40, 113.72, 106.11, 34.66; HRMS (ESI) m / z [M+H] + Calculate for C 12 H 10 ClN5O: 276.0647, found: 276.0645.

[0062] Example 5

[0063] The pyrazolopyrimidinone derivative I5 containing an aniline unit of the present invention, R1 = 2-Br, has a chemical name of 1-methyl-5-(2-bromoanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, an English name of 1-methyl-5-(2-bromophenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, and has the following structural formula:

[0064]

[0065] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0066] As in the first step of Example 1.

[0067] Step 2: Synthesis of 5-amino-1-methyl-N′-(2-bromophenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B5)

[0068]

[0069] The process is the same as the second step of Example 1, except that 2-bromophenylhydrazine is used as the reactant.

[0070] Step 3: Synthesis of 1-methyl-5-(2-bromoanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I5)

[0071] The same as the third step of Example 1, except that 5-amino-1-methyl-N′-(2-bromophenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0072] The target compound I5 is a white solid; yield 51%; mp 205-207°C; 1 H NMR (400MHz, DMSO-d6) δ8.74 (s, 1H), 8.52 (s, 1H), 8.16 (s, 1H), 7.56 (d, J=7.7Hz, 1H ), 7.15 (t, J=7.6Hz, 1H), 6.82 (t, J=7.4Hz, 1H), 6.40 (d, J=8.0Hz, 1H), 3.97 (s, 3H); 13 C NMR (100MHz, DMSO-d6) δ155.66, 153.27, 151.18, 144.69, 135.10, 133.27, 129.08, 122.43, 113.84, 107.90, 106.13, 34.66; HRMS (ESI) m / z [M+H] + Calculate for C 12 H 10 BrN5O: 320.0142, found: 320.0143.

[0073] Example 6

[0074] The pyrazolopyrimidinone derivative I6 containing an aniline unit of the present invention, R1 = 2,4-di-Cl, has a chemical name of 1-methyl-5-(2,4-dichloroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, an English name of 1-methyl-5-(2,4-dichlorophenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, and has the following structural formula:

[0075]

[0076] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0077] As in the first step of Example 1.

[0078] Step 2: Synthesis of 5-amino-1-methyl-N′-(2,4-dichlorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B6)

[0079]

[0080] The process is the same as the second step of Example 1, except that 2,4-dichlorophenylhydrazine is used as the reactant.

[0081] Step 3: Synthesis of 1-methyl-5-(2,4-dichloroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I6)

[0082] The process is the same as the third step of Example 1, except that 5-amino-1-methyl-N′-(2,4-dichlorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0083] The target compound I6 is a white solid; yield 56%; mp 217-219°C; 1 H NMR (400MHz, DMSO-d6) δ9.12 (s, 1H), 8.53 (s, 1H), 8.16 (s, 1H), 7.56 (d, J = 2.2Hz, 1H), 7.17 (dd, J = 8.8, 2.1Hz, 1H), 6.51 (d, J = 8.8Hz, 1H), 3.97 (s, 3H); 13 C NMR (100MHz, DMSO-d6) δ155.58, 153.10, 151.14, 142.91, 135.14, 129.35, 128.42, 124.68, 119.05, 115.13, 106.06, 34.66; HRMS (ESI) m / z [M+H] + Calculate for C 12 H9Cl2N5O: 310.0257, found: 310.0257.

[0084] Example 7

[0085] The pyrazolopyrimidinone derivative I7 containing an aniline unit of the present invention, R1 = 2-F, has a chemical name of 1-methyl-5-(2-fluoroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, an English name of 1-methyl-5-(2-fluorophenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, and has the following structural formula:

[0086]

[0087] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0088] As in the first step of Example 1.

[0089] Step 2: Synthesis of 5-amino-1-methyl-N′-(2-fluorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B7)

[0090]

[0091] The same as the second step of Example 1, except that 2-fluorophenylhydrazine is used as the reactant.

[0092] Step 3: Synthesis of 1-methyl-5-(2-fluoroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I7)

[0093] The same as the third step of Example 1, except that 5-amino-1-methyl-N′-(2-fluorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0094] The target compound I7 is a white solid; yield 51%; mp 235-237°C; 1 H NMR (400MHz, DMSO-d6) δ9.23 (s, 1H), 8.57 (s, 1H), 8.16 (s, 1H), 7.24-7.15 (m, 1H), 6. 98 (t, J=7.5Hz, 1H), 6.88 (dd, J=12.5, 6.4Hz, 1H), 6.56 (t, J=7.9Hz, 1H), 3.97 (s, 3H); 13 C NMR (100MHz, DMSO-d6) δ155.82, 153.25, 150.98 (d, J=240.1Hz), 135.52 (d, J=10.8Hz), 135.04, 125.26 (d, J= 3.0Hz), 121.62 (d, J=6.9Hz), 115.95 (d, J=17.8Hz), 114.96 (d, J=2.5Hz), 106.05, 34.63; HRMS (ESI) m / z [M+H] + Calculate for C 12 H 10 FN5O: 260.0942, found: 260.0943.

[0095] Example 8

[0096] The pyrazolopyrimidinone derivative I8 containing an aniline unit of the present invention, R1 = 2-Me, has a chemical name of 1-methyl-5-(2-methylanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, an English name of 1-methyl-5-(2-methylphenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, and has the following structural formula:

[0097]

[0098] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0099] As in the first step of Example 1.

[0100] Step 2: Synthesis of 5-amino-1-methyl-N′-(2-methylphenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B8)

[0101]

[0102] The same as the second step of Example 1, except that 2-methylphenylhydrazine is used as the reactant.

[0103] Step 3: Synthesis of 1-methyl-5-(2-methylanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I8)

[0104] The same as the third step of Example 1, except that 5-amino-1-methyl-N′-(2-methylphenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0105] The target compound I8 is a white solid; yield 72%; mp 222-224°C; 1 H NMR (400MHz, DMSO-d6) δ8.65 (s, 1H), 8.51 (s, 1H), 8.16 (s, 1H), 7.12 (d, J=7.3Hz, 1H), 6.9 7 (t, J=7.6Hz, 1H), 6.78 (t, J=7.3Hz, 1H), 6.17 (d, J=8.0Hz, 1H), 3.97 (s, 3H), 2.30 (s, 3H); 13 C NMR (100MHz, DMSO-d6) δ155.94, 153.35, 151.25, 145.65, 135.02, 130.94, 127.16, 123.17, 120.92, 111.37, 106.16, 34.64, 17.76; HRMS (ESI) m / z [M+H]+ Calculate for C 13 H 13 N5O: 256.1193, found: 256.1195.

[0106] Example 9

[0107] The pyrazolopyrimidinone derivative I9 ​​containing an aniline unit of the present invention, R1 = 3,5-di-Cl, has a chemical name of 1-methyl-5-(3,5-dichloroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, an English name of 1-methyl-5-(3,5-dichlorophenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, and has the following structural formula:

[0108]

[0109] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0110] As in the first step of Example 1.

[0111] Step 2: Synthesis of 5-amino-1-methyl-N′-(3,5-dichlorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B9)

[0112]

[0113] The process is the same as the second step of Example 1, except that 3,5-dichlorophenylhydrazine is used as the reactant.

[0114] Step 3: Synthesis of 1-methyl-5-(3,5-dichloroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I9)

[0115] The process is the same as the third step of Example 1, except that 5-amino-1-methyl-N′-(3,5-dichlorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0116] The target compound I9 is ​​a white solid; yield 55%; mp 237-239°C; 1 H NMR (400MHz, DMSO-d6) δ9.66 (s, 1H), 8.47 (s, 1H), 8.15 (s, 1H), 7.01 (s, 1H), 6.72 (d, J=1.6Hz, 2H), 3.95 (s, 3H); 13CNMR (100MHz, DMSO-d6) δ155.82, 152.86, 151.23, 150.69, 135.18, 135.13, 120.09, 111.76, 106.11, 34.64; HRMS (ESI) m / z[M+H] + Calculate for C 12 H9Cl2N5O: 310.0257, found: 310.0259.

[0117] Example 10

[0118] The pyrazolopyrimidone derivative containing aniline unit I of the present invention 10 , R1=2,4-di-Me, the chemical name is 1-methyl-5-(2,4-dimethylanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, the English name is 1-methyl-5-(2,4-dimethylphenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, the structural formula is as follows:

[0119]

[0120] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0121] As in the first step of Example 1.

[0122] Step 2: 5-amino-1-methyl-N′-(2,4-dimethylphenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B 10 )

[0123]

[0124] The process is the same as the second step of Example 1, except that 2,4-dimethylphenylhydrazine is used as the reactant.

[0125] Step 3: 1-methyl-5-(2,4-dimethylanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I 10 )

[0126] The process is the same as the third step of Example 1, except that 5-amino-1-methyl-N′-(2,4-dimethylphenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0127] Target compound I 10 White solid; yield 58%; mp 220-222℃;1 H NMR (400MHz, DMSO-d6) δ8.52 (s, 1H), 8.49 (s, 1H), 8.15 (s, 1H), 6.94 (s, 1H), 6.7 7 (d, J=8.1Hz, 1H), 6.07 (d, J=8.1Hz, 1H), 3.96 (s, 3H), 2.26 (s, 3H), 2.16 (s, 3H); 13 C NMR (100MHz, DMSO-d6) δ155.97, 153.31, 151.25, 143.34, 134.96, 131.60, 129. 67, 127.39, 123.29, 111.72, 106.17, 34.62, 20.51, 17.68; HRMS(ESI)m / z[M+H] + Calculate for C 14 H 15 N5O: 270.1349, found: 270.1351.

[0128] Embodiment 11

[0129] The pyrazolopyrimidone derivative containing aniline unit I of the present invention 11 , R1=2,4-di-F, the chemical name is 1-methyl-5-(2,4-difluoroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, the English name is 1-methyl-5-(2,4-difluorophenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, the structural formula is as follows:

[0130]

[0131] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0132] As in the first step of Example 1.

[0133] Step 2: 5-amino-1-methyl-N′-(2,4-difluorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B 11 )

[0134]

[0135] The process is the same as the second step of Example 1, except that 2,4-difluorophenylhydrazine is used as the reactant.

[0136] Step 3: 1-methyl-5-(2,4-difluoroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I 11 )

[0137] The same as the third step of Example 1, except that 5-amino-1-methyl-N′-(2,4-difluorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0138] Target compound I 11 White solid; yield 48%; mp 230-232°C; 1 H NMR (400MHz, DMSO-d6) δ9.19 (s, 1H), 8.57 (s, 1H), 8.14 (s, 1H), 7.27 (ddd, J=11.7 , 8.9, 2.6Hz, 1H), 6.88 (t, J=8.5Hz, 1H), 6.67 (td, J=9.3, 5.6Hz, 1H), 3.96 (s, 3H); 13 C NMR (100MHz, DMSO-d6) δ156.67 (d, J=239.6Hz), 155.85, 153.14, 151.17, 150.75 (d, J=243.9Hz), 135.01, 132.22 (dd, J=10.7, 3. 1Hz), 116.24 (dd, J=9.6, 3.5Hz), 111.66 (dd, J=22.0, 3.8Hz), 106.02, 104.72 (dd, J=26.7, 22.7Hz), 34.61; HRMS (ESI) m / z [M+H] + Calculate for C 12 H9F2N5O: 278.0848, found: 278.0850.

[0139] Example 12

[0140] The pyrazolopyrimidone derivative containing aniline unit I of the present invention 12 , R1=4-F, the chemical name is 1-methyl-5-(4-fluorophenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, the English name is 1-methyl-5-(4-fluorophenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, and the structural formula is as follows:

[0141]

[0142] Step 1: Synthesis of 5-amino-1-methyl-1H-1-pyrazole-4-carboxylic acid (Intermediate A)

[0143] As in the first step of Example 1.

[0144] Step 2: 5-amino-1-methyl-N′-(4-fluorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B 12 )

[0145]

[0146] The same as the second step of Example 1, except that 4-fluorophenylhydrazine is used as the reactant.

[0147] Step 3: 1-methyl-5-(4-fluoroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I 12 )

[0148] The same as the third step of Example 1, except that 5-amino-1-methyl-N′-(4-fluorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0149] Target compound I 12 White solid; yield 57%; mp 224-226℃; 1 H NMR (400MHz, DMSO-d6) δ9.27 (s, 1H), 8.51 (s, 1H), 8.15 (s, 1H), 7.07-7.00 (m, 2H), 6.72-6.67 (m, 2H), 3.96 (s, 3H); 13 C NMR (100MHz, DMSO-d6) δ157.36 (d, J=235.8Hz), 156.08, 153.22, 151.25, 144.57 (d, J=2.0H z), 134.99, 116.05 (d, J=22.6Hz), 114.83 (d, J=7.8Hz), 106.21, 34.60; HRMS (ESI) m / z [M+H] + Calculate for C 12 H 10 FN5O: 260.0942, found: 260.0944.

[0150] Example 13

[0151] The pyrazolopyrimidone derivative containing aniline unit I of the present invention 13, R1 = 2-OMe, chemical name is 1-methyl-5-(2-methoxyphenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, English name is 1-methyl-5-(2-methoxyphenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, the structural formula is as follows:

[0152]

[0153] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0154] As in the first step of Example 1.

[0155] Step 2: 5-amino-1-methyl-N′-(2-methoxyphenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B 13 )

[0156]

[0157] The process is the same as the second step of Example 1, except that 2-methoxyphenylhydrazine is used as the reactant.

[0158] Step 3: 1-methyl-5-(2-methoxyanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I 13 )

[0159] The same as the third step of Example 1, except that 5-amino-1-methyl-N′-(2-methoxyphenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0160] Target compound I 13 White solid; yield 56%; mp 219-221°C; 1 H NMR (400MHz, DMSO-d6) δ8.55 (s, 1H), 8.48 (s, 1H), 8.12 (s, 1H), 7.01-6.98 (m, 1H), 6.84 (td, J= 7.9, 1.4Hz, 1H), 6.73 (t, J=7.6Hz, 1H), 6.29 (dd, J=7.8, 1.2Hz, 1H), 3.96 (s, 3H), 3.87 (s, 3H); 13C NMR (100MHz, DMSO-d6) δ155.82, 153.50, 151.23, 147.29, 136.75, 134.94, 121.33, 121.21, 112.24, 111.50, 106.21, 56.05, 34.60; HRMS (ESI) m / z [M+H] + Calculate for C 13 H 13 N5O2: 272.1142, found: 272.1144.

[0161] Embodiment 14

[0162] The pyrazolopyrimidone derivative containing aniline unit I of the present invention 14 , R1=3-Cl, the chemical name is 1-methyl-5-(3-chlorophenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, the English name is 1-methyl-5-(3-chlorophenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, and the structural formula is as follows:

[0163]

[0164] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0165] As in the first step of Example 1.

[0166] Step 2: 5-amino-1-methyl-N′-(3-chlorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B 14 )

[0167]

[0168] The process is similar to the second step of Example 1, except that 3-chlorophenylhydrazine is used as the reactant.

[0169] Step 3: 1-methyl-5-(3-chloroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I 14 )

[0170] The same as the third step of Example 1, except that 5-amino-1-methyl-N′-(3-chlorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0171] Target compound I 14 White solid; yield 56%; mp 229-231°C;1 H NMR (400MHz, DMSO-d6) δ9.49 (s, 1H), 8.49 (s, 1H), 8.15 (s, 1H), 7.21 (t, J=8.1Hz, 1H), 6.8 9 (dd, J=7.9, 1.1Hz, 1H), 6.70 (t, J=1.8Hz, 1H), 6.62 (dd, J=8.2, 1.6Hz, 1H), 3.96 (s, 3H); 13 C NMR (100MHz, DMSO-d6) δ155.93, 153.12, 151.22, 149.76, 135.07, 134.21, 131.29, 120.68, 112.83, 111.90, 106.14, 34.64; HRMS (ESI) m / z [M+H] + Calculate for C 12 H 10 ClN5O: 276.0647, found: 276.0649.

[0172] Embodiment 15

[0173] The pyrazolopyrimidone derivative containing aniline unit I of the present invention 15 , R1=3-Me, the chemical name is 1-methyl-5-(3-methylphenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, the English name is 1-methyl-5-(3-methylphenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, and the structural formula is as follows:

[0174]

[0175] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0176] As in the first step of Example 1.

[0177] Step 2: 5-amino-1-methyl-N′-(3-methylphenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B 15 )

[0178]

[0179] The same as the second step of Example 1, except that 3-methylphenylhydrazine is used as the reactant.

[0180] Step 3: 1-methyl-5-(3-methylanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I 15 )

[0181] The same as the third step of Example 1, except that 5-amino-1-methyl-N′-(3-methylphenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0182] Target compound I 15 White solid; yield 59%; mp 223-225℃; 1 H NMR (400MHz, DMSO-d6) δ9.19 (s, 1H), 8.48 (s, 1H), 8.14 (s, 1H), 7.07 (t, J=7.6H z, 1H), 6.66 (d, J=7.4Hz, 1H), 6.44 (d, J=7.7Hz, 2H), 3.96 (s, 3H), 2.19 (s, 3H); 13 C NMR (100MHz, DMSO-d6) δ156.10, 153.35, 151.24, 148.19, 138.91, 135.00, 129.44, 121.85, 113.59, 110.49, 106.22, 34.62, 21.61; HRMS (ESI) m / z [M+H] + Calculate for C 13 H 13 N5O: 256.1193, found: 256.1195.

[0183] Example 16

[0184] The pyrazolopyrimidone derivative containing aniline unit I of the present invention 16 , R1=34-di-Cl, the chemical name is 1-methyl-5-(3,4-dichlorophenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, the English name is 1-methyl-5-(3,4-dichlorophenylamino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, the structural formula is as follows:

[0185]

[0186] Step 1: Synthesis of 5-amino-1-methyl-1H-pyrazole-4-carboxylic acid (Intermediate A)

[0187] As in the first step of Example 1.

[0188] Step 2: 5-amino-1-methyl-N′-(3,4-dichlorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide (Intermediate B 16 )

[0189]

[0190] The process is the same as the second step of Example 1, except that 3,4-dichlorophenylhydrazine is used as the reactant.

[0191] Step 3: 1-methyl-5-(3,4-dichloroanilino)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (target compound I 16 )

[0192] The same as the third step of Example 1, except that 5-amino-1-methyl-N′-(3,4-dichlorophenyl)-1H-pyrazole-4-carboxylic acid hydrazide is used as the reactant.

[0193] Target compound I 16 White solid; yield 71%; mp 262-264°C; 1 H NMR (400MHz, DMSO-d6) δ9.57 (s, 1H), 8.49 (s, 1H), 8.15 (s, 1H), 7.41 (d, J = 8.8Hz, 1H), 6.92 (d, J = 2.6Hz, 1H), 6.69 (dd, J = 8.8, 2.7Hz, 1H), 3.95 (s, 3H); 13 C NMR (100MHz, DMSO-d6) δ155.87, 152.98, 151.22, 148.37, 135.11, 132.03, 131.40, 122.34, 114.73, 113.75, 106.12, 34.64; HRMS (ESI) m / z [M+H] + Calculate for C 12 H9Cl2N5O: 310.0257, found: 310.0260.

[0194] Application performance testing

[0195] Test method for inhibiting activity against plant pathogenic fungi:

[0196] The mycelium growth rate method was used to determine the inhibitory effect of the target molecule on wheat fusarium graminearum, rice sheath blight (Rhizoctonia solanis), tomato gray mold (Botrytis cinerea) and tomato early blight (Alternaria solani) at a concentration of 50 μg / mL. Add 200 g of potato slices to 800 mL of boiling secondary water, continue boiling until it can be easily broken with a glass rod, filter with gauze, and set aside the filtrate. Measure another 200 mL of secondary water, add 20 g of agar powder, stir evenly, add the filtrate obtained in the previous step, add secondary water to 1 L, boil, add 20 g of glucose, stir evenly, and dispense into conical flasks to obtain PDA culture medium. After the newly prepared PDA medium is sterilized at 121°C for 20 minutes, use a 200μL pipette to measure 100μL of drug-containing DMSO and add it to 45mL of sterilized and uncooled PDA medium. After shaking, transfer the drug-containing medium evenly to three disposable culture dishes and cool naturally under ultraviolet light. After cooling, inoculate a 5mm diameter bacterial cake into the center of the disposable culture dish, and then transfer it to a 25°C constant temperature incubator and culture it upside down under dark conditions. When the colony diameter (C) of the blank control grows to 70-80mm, measure the colony diameter (T) of each treatment group using the cross method. Then calculate the inhibition rate (I) of the corresponding treatment group on plant pathogenic fungi based on the colony diameters obtained in the blank group and the drug group. The calculation formula is as follows:

[0197]

[0198] The target molecules I1-I were determined by the mycelial growth rate method using the agricultural fungicide oxazolidinone as a positive control. 16 The inhibitory activity of the agent against plant pathogenic fungi such as wheat fusarium rust, rice sheath blight, tomato gray mold and tomato early blight at a concentration of 50 μg / mL is shown in Table 1.

[0199] Table 1 Compounds I1-I 16 Inhibitory activity against plant pathogenic fungi at a concentration of 50 μg / mL

[0200] Compound Gibberella graminearum Rhizoctonia solani Tomato Botrytis cinerea Tomato early blight <![CDATA[I1]]> 5.95±1.18 4.56±1.01 7.15±0.46 6.16±0.49 <![CDATA[I2]]> 39.05±1.22 53.19±2.34 17.84±0.55 30.57±0.88 <![CDATA[I3]]> 15.29±1.09 6.97±1.53 11.02±1.29 8.27±0.88 <![CDATA[I4]]> 81.52±2.55 2.03±0.84 53.34±0.74 5.79±1.00 <![CDATA[I5]]> 83.11±0.45 1.17±1.27 70.98±2.51 4.72±0.58 <![CDATA[I6]]> 68.23±1.46 52.98±2.77 58.41±1.01 30.12±1.05 <![CDATA[I7]]> 20.75±2.06 5.53±0.82 7.54±1.18 3.81±0.65 <![CDATA[I8]]> 30.38±2.24 15.36±1.31 21.02±0.97 1.05±0.86 <![CDATA[I9]]> 7.64±1.03 39.22±3.33 10.88±1.43 3.40±0.46 <![CDATA[I 10 ]]> 13.06±0.75 7.66±1.31 5.76±1.43 4.58±0.44 <![CDATA[I 11 ]]> 8.91±1.05 10.35±1.16 7.10±1.29 8.71±0.44 <![CDATA[I 12 ]]> 35.67±2.16 23.68±2.11 12.25±0.99 23.02±0.35 <![CDATA[I 13 ]]> 91.63±1.65 20.02±2.32 12.25±0.99 1.00±0.63 <![CDATA[I 14 ]]> 10.02±1.09 12.37±1.12 11.36±1.22 6.65±0.42 <![CDATA[I 15 ]]> 8.14±0.54 16.23±0.99 17.54±1.22 6.37±0.28 <![CDATA[I 16 ]]> 25.86±1.52 55.58±1.25 12.63±0.23 14.22±0.33 Methadone 48.66±3.15 36.90±2.30 64.01±2.49 37.05±2.17

[0201] As shown in Table 1, some target molecules showed significant antibacterial activity against wheat fusarium, rice sheath blight and tomato gray mold at 50 μg / mL. 13At 50 μg / mL, it can significantly inhibit the growth of wheat fusarium mycelium, and the inhibition rates are 81.52%, 83.11%, 68.23% and 91.63%, respectively, which is better than the inhibition rate of oxadipazole on wheat fusarium at this concentration (48.66%). 16 At 50 μg / mL, they also showed significant inhibitory activity against rice sheath blight, with inhibition rates of 53.19%, 52.98% and 55.58%, respectively. In addition, target molecules I4, I5 and I6 also showed significant inhibitory activity against tomato gray mold at 50 μg / mL, with inhibition rates of 53.34%, 70.98% and 58.41%, respectively. It is worth noting that compound I 13 The inhibition rate against wheat fusarium rust reached 91.63% at 50 μg / mL, and it has the potential to be developed as a potential agricultural fungicide.

Claims

1. A pyrazolopyrimidone derivative containing an aniline unit and a preparation method and application thereof, characterized in that: The structural formula is shown in I: Among them, R 1 is selected from hydrogen, 2-methyl, 3-methyl, 4-methyl, 2,4-dimethyl, 2-chloro, 3-chloro, 4-chloro, 2,4-dichloro, 3,4-dichloro, 3,5-dichloro, 2-fluoro, 4-fluoro, 2,4-difluoro, 2-bromo or 2-methoxy.

2. The method for preparing the pyrazolopyrimidone derivative containing aniline unit according to claim 1, characterized in that: After 5-amino-1-methylpyrazole-4-carboxylic acid ethyl ester is hydrolyzed with 5% sodium hydroxide solution, it reacts with substituted phenylhydrazine in dichloromethane containing triethylamine and TBTU. The obtained hydrazide molecule undergoes cyclization reaction with triethyl orthoformate in hot ethanol to obtain the pyrazolopyrimidone derivative I containing aniline unit. The synthetic route is as follows:

3. Use of the pyrazolopyrimidone derivatives containing aniline units according to claim 1 in preventing and controlling plant pathogenic fungal diseases.

4. The use according to claim 3, characterized in that: The plant pathogenic fungi are wheat fusarium rust, rice sheath blight or tomato gray mold.

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