Preparation method and application of alpha-phosphoramidate compound containing isopropanolamine substructure

Abstract

The invention relates to a preparation method and application of an alpha-phosphoramidate compound containing an isopropanolamine substructure. The compound has a structure as shown in a general formula (I): # imgabs0 #, alpha-phosphoramidate is taken as a lead skeleton, isopropanolamine-containing fragments are introduced into the system, a series of alpha-phosphoramidate compounds containing isopropanolamine fragments are synthesized, and the compounds have a significant effect on plant virus diseases, plant pathogenic bacterial diseases such as tobacco mosaic virus, plant pathogenic bacterial diseases such as tobacco mosaic virus, plant pathogenic bacterial diseases, plant pathogenic bacterial diseases, plant pathogenic bacterial diseases and the like. The compound has an excellent inhibition effect on virus, cucumber mosaic virus, rice bacterial leaf blight, rice bacterial leaf streak, citrus canker pathogen, kiwi canker pathogen, tomato canker pathogen and the like.

Description

Technical Field

[0001] The invention relates to the technical field of pharmaceutical chemistry, in particular to an alpha-phosphoramidate compound containing an isopropanolamine substructure and a preparation method and application thereof. Background Art

[0002] Plant diseases are one of the main factors affecting global agricultural production, seriously affecting the yield and quality of agricultural products, not only causing great economic losses, but also threatening human health. For example, tobacco mosaic virus, cucumber mosaic virus, potato virus Y, rice bacterial leaf blight, rice bacterial streak disease, citrus canker, kiwi canker, tobacco bacterial wilt, etc., break out to varying degrees every year, causing huge economic losses to farmers. The long-term use of traditional fungicides, such as thiophanate-methyl, chlorothiazide, streptomycin sulfate, etc., not only increases the drug resistance of plant pathogens, but also has a harmful effect on the ecological environment and plant safety. Therefore, there is an urgent need to develop new pesticides with high activity and high selectivity.

[0003] According to literature reports, α-aminophosphoric acid ester derivatives exhibit a broad spectrum of biological activities, such as antibacterial, anti-insecticide, herbicide, plant growth regulation, anti-tumor, anti-inflammatory, etc. According to the previous work of this research group, α-aminophosphoric acid ester compounds show good anti-plant pathogenic activity.

[0004] In order to find an active compound with high efficiency in killing bacteria, the present invention uses α-phosphoramidate as a matrix, connects it to propylene oxide, and then opens the ring to obtain the target α-phosphoramidate compound containing an isopropanolamine fragment. A series of α-phosphoramidate compounds containing an isopropanolamine fragment with novel structures are prepared, and their biological activities are tested, which provides an important scientific basis for the research and development and creation of new pesticides.

[0005] The research progress of biological activity of α-phosphoramidate compounds is as follows:

[0006] In 2017, Zhang et al. [Zhang, GP; Hao, GF; Pan, JK; Zhang, J.; Hu, DY; Song, BA; Asymmetric synthesis and bioselective activities of α-amino-phosphonatesbased on the dufulin motif. J. Agr. Food Chem., 2016, 64(21), 4207-4213] designed, synthesized and studied the bioactivity of chiral phosphonates and chalcone derivatives based on natural products. The target compound (R)-Ib was tested by the half-leaf spot method and its therapeutic activity against cucumber mosaic virus at a concentration of 500 μg / ml was 72.3%.

[0007] In 2023, Yang et al. [Yang, XC; Zeng, CM; Avula, SR; Peng, XM,: Geng RX; Zhou, CH; Novel coumarin aminophosphonates as potential multitargetingantibacterial agents against Staphylococcus aureu. Eur. J. Med. Chem., 2023, 245, 114891] synthesized a series of compounds with α-aminophosphonates as the parent structure, among which compound 6f had a good inhibitory effect on Staphylococcus aureus at a concentration of 0.5 μg / mL. 6f can eradicate Staphylococcus aureus biofilm, thereby inhibiting the infection of Staphylococcus aureus.

[0008] In 2018, Zhang et al. [Zhang, Y.; Tangadanchu, VKR; Cheng, Y.; Yang, RG; Lin, JM; Zhou, CH Potential antimicrobial isopropanol-conjugated carbazole azoles as dual targeting inhibitors of enterococcus faecalis. ACS Med. Chem. Lett., 2018, 9, 244-249] designed and synthesized a series of isopropanolamine compounds and tested their activity against Enterococcus faecalis. The results showed that compound 10 had high activity (MIC = 2 μg / mL). Summary of the invention

[0009] One of the purposes of the present invention is to provide an α-aminophosphoric acid ester compound containing an isopropanolamine fragment or a salt thereof or a solvate thereof.

[0010] Another object of the present invention is to provide an intermediate compound for preparing the above compound or its salt or solvate and a preparation method thereof.

[0011] Another object of the present invention is to provide a composition containing the above compound or its salt or solvate.

[0012] Another object of the present invention is to provide the use of the above compound or its salt or solvate, or the composition.

[0013] Another object of the present invention is to provide a method for controlling agricultural pests and diseases using the above compound or its salt or solvate, or the composition.

[0014] To achieve the above object, the present invention adopts the following technical solutions:

[0015] An α-phosphoramidate compound containing an isopropanolamine substructure or a salt thereof or a solvate thereof, wherein the compound has a structure as shown in general formula (I):

[0016]

[0017] Wherein, R1, R2, R3, R4, and R5 are each independently selected from hydrogen, any substituted or unsubstituted alkyl, any substituted or unsubstituted alkoxy, any substituted or unsubstituted cycloalkyl, any substituted or unsubstituted aryl, any substituted or unsubstituted heteroaryl, any substituted or unsubstituted benzyl, any substituted or unsubstituted α-methyl-benzyl, any substituted or unsubstituted N-methyl-benzyl, and N-ethyl-benzyl.

[0018] The present invention also provides an intermediate compound for preparing the isopropanolamine-containing α-phosphoramidate compound or its salt or solvate:

[0019]

[0020] The present invention also provides a method for preparing the isopropanolamine compound containing α-phosphoramidate or its salt or solvate, comprising the following steps:

[0021]

[0022] The present invention also provides a method for preventing and controlling agricultural pests and diseases, wherein the compound or its salt or solvate, or the composition acts on harmful substances or their living environment; preferably, the agricultural pests and diseases are plant viral diseases, plant bacterial or fungal diseases; more preferably, the agricultural pests and diseases are cucumber mosaic virus, tobacco mosaic virus, potato virus Y, rice bacterial leaf blight, rice bacterial leaf streak disease, tobacco bacterial wilt, cucumber bacterial leaf blight, konjac bacterial leaf blight, citrus canker, grape canker, tomato canker, kiwi canker, apple canker, cucumber gray mold, pepper wilt pathogen, rapeseed sclerotinia, wheat fusarium head blight, and potato late blight.

[0023] The present invention also provides a method for protecting plants from agricultural pests and diseases, which comprises a method step in which the plants are contacted with the compound or its salt or solvate, or the composition.

[0024] Combinations of substituents and variables are permissible only if such combinations result in stable compounds or useful synthetic intermediates. A stable compound or stable structure implies that the compound is sufficiently stable to be isolated in a useful degree of purity from a reaction mixture, and then formulated into an efficacious therapeutic agent.

[0025] By adopting the above technical scheme, the present invention uses aldehyde and amine as starting materials to synthesize a series of α-phosphoramidate compounds containing isopropanolamine fragments, and it is found that the compounds have good inhibitory effects on pathogenic plant viral diseases and pathogenic bacteria, and have good inhibitory effects on tobacco mosaic virus (Tobacco mosaic virus, TMV), plant pathogenic bacteria [such as rice bacterial leaf blight pathogen (Xanthomonasoryzae pv.oryzae, Xoo), rice bacterial leaf streak pathogen (Xanthomonasoryzae pv.oryzicola, Xoc), citrus canker pathogen (Xanthomonas axonopodis pv.citri, Xac) and kiwi canker pathogen (Pseudomonassyringae pv.actinidiae, Psa), tomato canker pathogen (Clavibactermichiganensis subsp.michiganensis, Cmm), etc.], providing an important scientific basis for the research and development and creation of new pesticides. Example

[0026] The present invention is further described below by way of examples. It should be understood that the methods described in the examples of the present invention are only used to illustrate the present invention, rather than to limit the present invention, and simple improvements to the preparation methods of the present invention under the premise of the concept of the present invention belong to the scope of protection claimed in the present invention. All raw materials and solvents used in the examples are commercially available products of corresponding purity.

[0027] Example 1: Preparation of intermediate diethyl (((2,4-dichlorophenyl)amino)(4-hydroxyphenyl)methyl)phosphonate

[0028] 2,4-Dichloroaniline (3.09mmol) and p-hydroxybenzaldehyde (3.09mmol) and diethyl phosphite (3.70mmol) were added to a 50mL round-bottom flask, stirred at 80°C for 10 minutes to terminate the reaction; ethyl acetate (30mL) was used for extraction, and then washed with saturated NH4Cl aqueous solution (3×20mL), dried over anhydrous Na2SO4, desolventized, and column chromatography (PE:EA=3:1, V / V) was performed to obtain a white solid with a yield of 49%. Its NMR data are: 1H NMR (400MHz, DMSO,) δ7.41 (s, 2H, Aniline-H), 7.23 (d, J=6.9Hz, 2H, Benzene-H), 7.11 (d, J=8.0Hz, 1H, A niline-H), 6.79 (d, J=8.6Hz, 1H, Aniline-H), 6.73 (d, J=7.5Hz, 2H, Benzene-H), 5.29 (t, J=8.3Hz, 1H, N H ), 5.09 (dd, J=23.5, 8.8Hz, 1H, C H P), 4.16-3.62 (m, 4H, 2OC H2 CH3), 1.20 (t, 3H, OCH2C H3 ), 1.04(t, J=6.4Hz, 3H, OCH2C H3 ); 31 P NMR (162MHz, DMSO) δ22.68 (s):

[0029] Example 2: Preparation of the intermediate diethyl (((2,4-dichlorophenyl)amino)(4-(2-epoxypropyl)phenyl)phosphonate)

[0030] The intermediate diethyl (((2.4-dichlorophenyl)amino)(4-hydroxyphenyl)methyl)phosphonate (4.94mmol), KOH (5.94mmol) and epibromopropane (7.42mmol) were added to a 25mL round-bottom flask and reacted at room temperature for 6 hours. After solvent removal and column chromatography (CH2Cl2:CH3OH=30:1, V / V), a light yellow oil was obtained with a yield of 66%. Its NMR data are: 1 H NMR (400MHz, CDCl3, ppm) δ7.34 (dd, J=8.7, 2.3Hz, 2H, Benzene-H), 7.25 (d, J=2.5Hz, 1H, Aniline-H), 6.94 (dd, J=8.7, 2.3Hz, 1H, Aniline -H), 6.89 (d, J=8.6Hz, 2H, Benzene-H), 6.36 (d, J=8.8Hz, 1H, Aniline-H), 5.33 (dd, J=10.1, 7.5Hz, 1H.NH), 4.67 (dd, J=23.7, 7.4Hz, 1H, C H P), 4.25-4.19 (m, 1H, OC H 2CH), 4.16-3.87 (m, 4H, 2OC H2 CH3), 3.84-3.73 (m, 1H, OC H2CH), 3.34 (dd, J=6.1, 3.3Hz, 1H, C H O), 2.90 (t, J=4.5Hz, 1H, Epoxy-H), 2.77-2.71 (m, 1H, Epoxy-H), 1.28 (t, J=7.1Hz, 3H, OCH2C H3 ), 1.18(t, J=7.1Hz, 3H, OCH2C H3 ).

[0031] Example 3: Preparation of target compound 2 diethyl (((2,4-dichlorophenyl)amino)(4-(2-hydroxyethyl)amino)propoxy)methyl)phosphonate

[0032] Dissolve diethyl (((2.4-dichlorophenyl)amino)(4-(2-epoxypropyl)phenyl)phosphonate (0.65mmol), 2-hydroxyethylamine (1.30mmol) and K2CO3 (0.65mmol) in 3ml isopropanol and put into a 15mL reaction bottle. Stir and react at 50℃ for 6h. Follow up with TLC. After the reaction is complete, add water to quench the reaction. Then extract with ethyl acetate (50mL). Take the organic phase, dry it with anhydrous Na2SO4, remove the solvent, and then perform column chromatography (CH2Cl2:CH3OH=30:1, V / V) to obtain a yellow solid with a yield of 82%. Its NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ7.32 (d, J=6.5Hz, 2H, Benzene-H), 7.24 (d, J=2.4Hz, 1H, Aniline-H), 6.92 (dd, J=8.7, 2.4Hz, 1H, Aniline-H), 6.87 (d, J=8.6Hz, 2H, Benzene-H), 6.36 (d, J=8.9Hz, 1H, Aniline-H), 5.31 (dd, J=10.2, 7.5Hz, 1H, N H PH), 4.67 (dd, J=23.7, 7.4Hz, 1H, PC H ), 4.15-4.02 (m, 7H, 2OC H2 CH3, OC H2 C H OH), 3.74 (t, 2H, NHCH2C H2 OH), 3.12-2.79 (m, 4H, C H2 NHC H2 ), 1.92(s,1H,CH2CHO H CH2), 1.28(t, J=7.1Hz, 3H, OCH2C H3), 1.18(t, J=7.1Hz, 3H, OCH2C H 3). 13 C NMR (101MHz, CDCl3) δ158.68, 141.29, 129.14, 129.09, 127.86, 127.42, 122.83, 120.66, 115.09, 113.57, 70.41, 67.55, 63.79 (dd, 2 J C -P=11.6, 7.1Hz), 59.79, 55.57 (d, 1 J C-P =152.6Hz), 51.68, 51.20, 16.64 (dd, 3 J C-P =13.8, 5.7 Hz); 31 P NMR (162MHz, CDCl3) δ21.50;

[0033] Example 4: Preparation of target compound 5 diethyl ((4-(3-(cyclopropylamino)-2-propoxy)phenyl)(((2,4-dichlorophenyl)amino)methyl)phosphate

[0034] Dissolve diethyl (((2.4-dichlorophenyl)amino)(4-(2-epoxypropyl)phenyl)phosphonate (0.65mmol), cyclopropylamine (1.30mmol) and K2CO3 (0.65mmol) in 3ml isopropanol and put into a 15mL reaction bottle. Stir and react at 50℃ for 6h. Follow up with TLC. After the reaction is complete, add water to quench the reaction. Then extract with ethyl acetate (50mL). Take the organic phase, dry it over anhydrous Na2SO4, remove the solvent, and obtain a light yellow oil by column chromatography (CH2Cl2:CH3OH=30:1, V / V). The yield is 86%. Its NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ7.33 (dd, J=8.8, 2.3Hz, 2H, Benzene-H), 7.25 (d, J=2.4Hz, 1H, Aniline-H), 6.94 (dd, J=8.7, 2.4H z, 1H, Aniline-H), 6.89 (d, J=8.5Hz, 2H. Benzene-H), 6.37 (d, J=8.8Hz, 1H, Aniline-H), 5.33 (dd, J=10.2, 7.4Hz, 1H, N H PH), 4.67 (dd, J=23.7, 7.4Hz, 1H, PC H ), 4.18-4.08 (m, 1H, C HOH), 4.08-3.90 (m, 5H, 2OC H2 CH3, OC H 2CHOH), 3.86-3.72(m, 1H, OC H 2CHOH), 2.90 (ddd, J=20.5, 12.3, 6.0Hz, 2H, NHC H2 ), 2.18 (ddd, J=10.1, 6.7, 3.6Hz, 1H, Cyclopropylamine-H), 1.28 (t, J=7.1Hz, 3H, OCH2C H3 ), 1.18(t, J=7.1Hz, 3H, OCH2C H 3), 0.47 (t, J=6.9Hz, 2H, Cyclopropylamine-H), 0.43-0.34 (m, 2H, Cyclopropylamine-H).; 13 C NMR (101MHz, CDCl3) δ158.62, 141.25, 128.83, 128.78, 127.60, 127.02, 122.53, 120.42, 114.83, 113.31, 70.47, 67.95, 63.47 (t, 2 J C -P=6.8Hz), 56.17(d, 1 J C -P=152.3Hz), 51.74, 30.49, 16.43 (dd, 3 J C -P=14.6, 5.6Hz), 6.07 (d, J=62.3Hz); 31 PNMR (162MHz, CDCl3) δ21.57;

[0035] Other target compounds were synthesized by using corresponding raw materials or substituents according to the steps of the above examples.

[0036] The structures, hydrogen, carbon, fluorine and phosphorus spectra of the synthesized isopropanolamine compounds containing α-aminophosphoric acid ester are shown in Table 1, and the physicochemical properties are shown in Table 2.

[0037] Table 1 Nuclear magnetic resonance hydrogen spectrum and carbon spectrum data of the compounds of the present application

[0038]

[0039]

[0040]

[0041]

[0042]

[0043]

[0044]

[0045]

[0046]

[0047]

[0048]

[0049]

[0050]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056]

[0057]

[0058] Table 2 Physicochemical properties of the compounds of this application

[0059]

[0060]

[0061]

[0062] Pharmacological Example 1:

[0063] EC 50Median effective concentration (MEC) is an important indicator for evaluating the sensitivity of plant pathogens to compounds. It is also an important parameter for setting compound concentration when studying the mechanism of action of target compounds. In the concentration gradient experiment, the two-fold dilution method was used to set 5 appropriate concentrations. Finally, the inhibition rate of the agent on plant pathogens and the concentration of the agent were converted into logarithmic values. The toxicity curve was obtained through regression analysis using SPSS software, and the EC was calculated. 50 .

[0064] The effective medium concentration EC of the target compound against plant pathogens was tested by turbidity method. 50 The test subjects were Xanthoceras oryzae (Xoo), Xanthoceras oryzae (Xoc), Xanthoceras citri (Xac), and Psa. DMSO was dissolved in the culture medium as a blank control. Xanthoceras oryzae (Xanthoceras oryzae in M210 solid culture medium) was placed in NB culture medium, and cultured in a constant temperature shaker at 28°C and 180rpm until the logarithmic growth phase for use; Xanthoceras citri (on M210 solid culture medium) was placed in NB culture medium, and cultured in a constant temperature shaker at 28°C and 180rpm until the logarithmic growth phase for use. 5 mL of NB liquid medium containing different concentrations (e.g., 100, 50, 25, 12.5, 6.25 μg / mL) of the drug (compound) was added to the test tube, and 40 μL of NB liquid medium containing plant pathogenic bacteria was added respectively. The mixture was shaken in a constant temperature shaker at 28°C and 180 rpm. The rice bacterial leaf blight pathogen was cultured for 36 hours, the rice bacterial leaf streak pathogen was cultured for 36 hours, the citrus canker pathogen was cultured for 48 hours, and the kiwi canker pathogen was cultured for 36 hours. The OD of the bacterial solution of each concentration was measured on a spectrophotometer. 595 The OD of the corresponding concentration of toxic sterile NB liquid culture medium was also measured. 595 value.

[0065] Corrected OD value = OD value of bacterial culture medium - OD value of sterile culture medium

[0066] Inhibition rate % = [(corrected control culture medium bacterial solution OD value - corrected toxic culture medium OD value) / corrected control culture medium bacterial solution OD value] × 100

[0067] The examples of the present invention are used to illustrate the technical solutions of the present invention, but the contents of the examples are not limited thereto. The experimental results of the target compounds are shown in Table 3.

[0068] Table 3 EC values ​​of the compounds of the present application against plant pathogenic bacteria 50

[0069]

[0070]

[0071]

[0072] As can be seen from Table 3, in the in vitro test, the target compounds showed good inhibitory activity against plant pathogenic bacteria (such as Xanthoceras oryzae, Xanthoceras oryzae, Xanthoceras citri and Xanthoceras kiwifruit). Most of the compounds containing benzyl and phenyl groups in their structures showed good inhibitory activity against Xanthoceras oryzae, Xanthoceras citri and Xanthoceras citri. 50 All of them are within 10, especially compound 29 has excellent activity against Xanthomonas oryzae, EC 50 At the same time, compound 25 also showed excellent inhibitory activity against rice bacterial leaf streak, EC 50 The compound 69 showed excellent inhibitory activity against citrus canker, EC 50 The compound 47 showed excellent inhibitory activity against kiwifruit canker, EC 50 It is 1.96 μg / mL. Therefore, it can be seen that this type of compound has great research prospects and can be used to prepare pesticides against plant pathogenic bacteria.

[0073] Pharmacological Example 2:

[0074] In order to study the anti-plant virus activity of some of the synthesized target compounds, we used the half-leaf spot method with Nicotiana tabacum as the model plant, cyfluthrin, ningnanmycin and ribavirin as control agents, and tested the biological activity of some of the synthesized compounds against tobacco mosaic virus (TMV) at a test compound concentration of 500 μg / mL.

[0075] The half-leaf spot method was used to test the inhibitory effect of the target compound on plant virus diseases. The test object was tobacco mosaic virus (TMV). Accurately weigh 2 mg of the target compound in a 15 mL centrifuge tube, and add 25 μL of DMSO to the centrifuge tube to fully dissolve the compound. Then use double distilled water (containing 1% Tween 80) to prepare a compound solution at a mass concentration of 500 mg / L. Take another 25 μL of 8% Ningnanmycin aqueous solution and 2 mg of fluazifop, and repeat the above method as a control.

[0076] The agent's living protective effect on plants infected with tobacco mosaic virus, select heart-leaf tobacco with consistent growth, remove the top and old leaves, leave 4-5 leaves of equivalent leaves per plant, use a brush to dip the test compound solution and gently apply it on the right side of the leaf, apply an equal dose of solvent on the left half of the leaf as a control, and then place the plant after the application in a light incubator for cultivation, and inoculate the virus on the whole leaf 24 hours later. First, sprinkle corundum evenly on the leaves, and then use a brush to dip the virus juice and inoculate the virus on the whole leaf. After 30 minutes, rinse off the corundum on the surface of the leaves and dry naturally. Place the plant in a constant temperature incubator with a temperature of 25±1℃ and a light of 10000Lux for moisturizing cultivation, and count the number of dead spots after the leaves have spots after 2-4 days. Repeat 3 times for each agent according to the above method, and calculate the inhibition rate.

[0077] The drug is used to treat the plants infected with tobacco mosaic virus in vivo. The action is to select the heart leaf tobacco with the same growth, remove the top and old leaves, and leave about 4-5 leaves of the same size for each plant. Sprinkle a layer of corundum on the surface of the leaves evenly, then dip a brush in the virus solution, and inoculate the virus on the leaf surface with a surface dish underneath along the direction of its branch veins. After 30 minutes, rinse off the corundum on the surface of the leaves and dry naturally. Dip a brush in the solution of the test compound and gently apply it on the right side of the leaf. Apply an equal amount of solvent to the left half of the leaf as a control, and then place the plant in a constant temperature incubator with a temperature of 25±1℃ and a light of 10000Lux for moisturizing and cultivation. After two to four days, count the number of dead spots after the leaves have spots. Repeat 3 times for each drug according to the above method, and calculate the inhibition rate.

[0078] Calculation of the necrosis inhibition rate (half-leaf necrosis method): After 2-4 days of greenhouse cultivation, when lesions are clearly seen, the number of lesions on the left and right halves of the leaves is recorded, and then the inhibition rate of the compound on the virus is calculated according to the following formula. The other half of the leaf is used as a control for each treatment group.

[0079] Inhibition rate % = [(number of shifts in the control group - number of shifts in the treatment group) / number of shifts in the control group] × 100

[0080] The examples of the present invention are used to illustrate the technical solutions of the present invention, but the contents of the examples are not limited thereto. The experimental results of the target compounds are shown in Table 4.

[0081] Table 4: The protective effect of some compounds of the present application on tobacco mosaic virus (TMV) at 500 μg / mL

[0082]

[0083]

[0084] As can be seen from Table 4, the target compounds showed good inhibitory activity against plant virus diseases (such as tobacco mosaic virus). In particular, compounds 16 and 33 had good therapeutic activity against tobacco mosaic virus, with inhibition rates of 75.4% and 68.4%, respectively. At the same time, compounds 27 and 66 had good protective activity against tobacco mosaic virus, with inhibition rates of 68.0% and 66.4%, respectively. It can be seen that this type of compound has good research prospects in resisting plant virus diseases and can be used to prepare pesticides against plant virus diseases.

[0085] Pharmacological Example 3:

[0086] Compound 26 showed the best activity against Xanthomonas oryzae (EC 50 The in vivo pot experiment of compound 29 on rice bacterial blight was conducted. The specific experimental steps are as follows:

[0087] Protective activity: Compound 26 and control drug thiophanate-methyl (20% preparation) were prepared into 200 μg / mL drug-containing solutions with less than 1% Tween20 solution, and two more 200 μg / mL solutions of compound 29 were prepared. The prepared solutions were sprayed on the surface of rice leaves that had grown for 8 weeks until droplets fell. After 24 hours, the leaves were sprayed with OD 595 = =0.6-0.8 range of rice bacterial blight pathogenic bacteria with scissors cut off the leaf tips, set the same amount of DMSO and bacterial leaves without adding drugs as controls, each treatment had three replicates, checked the disease situation after 14 days, recorded the length of the lesions and the total length of the rice leaves, and calculated the disease index and prevention effect.

[0088] First, the spot area and the entire leaf area of ​​each leaf were measured, and then the percentage of the entire spot area was measured to calculate the leaf area. Secondly, the leaves were classified according to the following grade standards: Grade 1, the spot area is less than 5% of the entire leaf area. Grade 3, the spot area accounts for 6-10% of the entire leaf area; Grade 5, the spot area accounts for 11-20% of the entire leaf area; Grade 7, the spot area accounts for 21-50% of the entire leaf area; Grade 9, the spot area accounts for more than 50% of the entire leaf area;

[0089] The calculation method of the disease index is as follows:

[0090] Disease index = ∑ (number of leaves in each level × corresponding level) / (total number of leaves × highest level)

[0091] The calculation method of protection effect is as follows:

[0092] Prevention effect % = (disease index of control group - disease index of treatment group) / disease index of control group × 100%

[0093] Therapeutic activity: Apply a solution of OD 595 = =0.6-0.8 range of rice bacterial blight pathogens with scissors; 24h later, the prepared liquid medicine was sprayed on the surface of rice leaves that had grown for 8 weeks until droplets fell, and an equal amount of DMSO and bacterial leaves without the addition of the agent were set as controls. Each treatment had three replicates. After 14 days, the disease situation was checked, the lesion length and total length of the rice leaves were recorded, and the disease index and prevention effect were calculated, and the calculation method was the same as above.

[0094] The examples of the present invention are used to illustrate the technical solutions of the present invention, but the contents of the examples are not limited thereto. The experimental results of the target compounds are shown in Table 5.

[0095] Table 5 Protective and therapeutic activities of the compounds of the present application against Xanthomonas oryzae

[0096]

[0097]

[0098] As can be seen from Table 4, in the in vivo test of the target compound 29 at a concentration of 200 μg / mL, compound 26 showed good therapeutic activity (30.76%) and protective activity (41.61%) against rice bacterial blight pathogen. It is better than the control drug thiophanate-methyl (therapeutic activity 26.57%; protective activity 38.92%). Therefore, it can be seen that this type of compound has great research prospects, and compound 26 is developed for the preparation of highly resistant pesticides to plant pathogenic bacteria.

[0099] Pharmacological Example 4:

[0100] Compound 62 showed the best activity against kiwifruit canker (EC 50 The compound 62 was tested on the leaves of kiwifruit canker pathogen in vivo. The specific experimental steps are as follows:

[0101] Protective activity: Select leaves of kiwifruit plants with consistent growth and darker color, use a hole puncher to take a disc, soak the leaf disc with a specific concentration of compound 62, use DMSO as a positive control, and thiophanate-methyl (TC) as a negative control; place the drug-treated plants in an incubator under light conditions and culture at 10-15℃ for 16h; then inoculate OD 595 = 0.08-0.1 of kiwifruit canker bacteria liquid, put it into the incubator, culture it in the dark, 10-15℃ for 8h, culture it for 5 days, and observe the spotting. The incubator conditions are changed to a day-night switching mode (daytime 16h, 15-20℃, humidity greater than 80%; nighttime 8h, 10-15℃, humidity greater than 80%).

[0102] The control efficiency is calculated as follows:

[0103] Control efficiency % = (chlorophyll content of water treatment group - chlorophyll content) / (chlorophyll content of water treatment group - chlorophyll content of CK treatment group)

[0104] Amount) × 100%

[0105] Therapeutic activity: Inoculation of OD on kiwi leaf discs 595 = 0.08-0.1 of kiwifruit canker bacteria liquid, placed indoors to dry, then placed in an incubator, cultured in dark conditions, 10-20°C for 24h; then a specific concentration of compound 62 was added to the wound, DMSO was used as a positive control, and TC was used as a negative control; then placed in an incubator, cultured in light conditions, 10-20°C for 24h; finally, the incubator conditions were changed to a day-night switching mode (10-15h during the day, 10-15°C, humidity greater than 80%; 10-15h at night, 10-15°C, humidity greater than 80%). The control efficiency calculation method is the same as above.

[0106] The examples of the present invention are used to illustrate the technical solutions of the present invention, but the contents of the examples are not limited thereto. The experimental results of the target compounds are shown in Table 6.

[0107] Table 6 Protective and therapeutic activities of the compounds of the present invention against kiwifruit canker pathogens

[0108]

[0109] As can be seen from Table 5, in the in vivo test, the target compound 62 at a concentration of 200 μg / mL showed good therapeutic activity (32.64%) and protective activity (52.98%) against kiwifruit canker pathogens. It was better than the control drug thiophanate-methyl (therapeutic activity 15.44%; protective activity 22.26%), indicating that this series of compounds has made great breakthroughs in in vitro and in vivo activities and can be used to prepare pesticides against plant pathogenic bacteria.

[0110] Pharmacological Example 5:

[0111] Compound 25 showed the best activity against Xanthomonas oryzae (EC 50 The experimental results were as follows:

[0112] Protective activity: Compound 25 and control drug thiophanate-methyl (20% content preparation) were prepared into 200 μg / mL drug-containing solutions with less than 1% Tween20 solution, and then a 200 μg / mL solution of compound 25 was prepared. The prepared drug solutions were sprayed on the surface of rice leaves that had grown for 8 weeks until droplets fell. After 24 hours, the leaves were inoculated at OD 1.50 at the top of the leaves. 595 = Rice bacterial leaf streak pathogen in the range of 0.6-0.8, with equal amounts of DMSO and bacterial solution without adding any agent as controls, and each treatment had three replicates. The disease situation was checked after 14 days, the length of the lesions on the rice leaves was recorded, and the disease situation and prevention efficacy were calculated.

[0113] First, measure the length of the spots on each leaf, measure the length of the diseased leaves, and calculate the prevention effect.

[0114] The calculation method of protection effect is as follows:

[0115] Prevention effect % = (lesion length of control group - lesion length of treatment group) / lesion length of control group × 100%

[0116] Curative activity: Inoculate OD at the top of leaves 595 = Rice bacterial leaf streak pathogen in the range of 0.6-0.8; 24h later, the prepared liquid medicine was sprayed on the surface of rice leaves that had grown for 8 weeks until droplets fell, and an equal amount of DMSO and bacterial solution without adding the medicine were set as controls. Each treatment had three replicates. After 14 days, the disease situation was checked, the length of the lesions on the rice leaves was recorded, and the disease index and prevention effect were calculated, and the calculation method was the same as above.

[0117] The examples of the present invention are used to illustrate the technical solutions of the present invention, but the contents of the examples are not limited thereto. The experimental results of the target compounds are shown in Table 7.

[0118] Table 7 Protective and therapeutic activities of the compounds of the present invention against rice bacterial leaf streak

[0119]

[0120] As can be seen from Table 6, in the in vivo test, compound 25 showed good therapeutic activity (42.46%) and protective activity (45.03%) against rice bacterial leaf streak, which was better than the control drug thiophanate-methyl (therapeutic activity 31.73%; protective activity 26.70%); thus, it can be seen that this type of compound has great research prospects and can be used to prepare pesticides with high resistance to plant pathogenic bacteria.

[0121] Pharmacological Example 6:

[0122] Compound 67 showed the best activity against X. citri 50The compound 67 was tested in a potted plant in vivo against citrus canker bacteria. The specific experimental steps are as follows:

[0123] Protective activity: Select citrus plants with uniform leaf growth, pierce holes in the leaves with a needle, add compound 67 at a concentration of 200 μg / ml, adsorb it on filter paper, and attach it to the wound. DMSO and clean water are used as negative controls, and thiophanate-methyl (TC) is used as a positive control. The leaves with the added drugs are naturally dried and then placed in an incubator under light conditions at 25°C for 24 hours. Then, OD 595 =0.01 of citrus canker bacteria solution, put it into an incubator, culture conditions are dark conditions, and culture at 20℃ for 24h; finally, the incubator conditions are changed to a day-night switching mode (10-15h during the day, 10-15℃, humidity greater than 80%; 10-15h at night, 10-15℃, humidity greater than 80%).

[0124] The control efficiency is calculated as follows:

[0125] Prevention effect % = (lesion length of control group - lesion length of treatment group) / lesion length of control group × 100%

[0126] Therapeutic activity: Punch holes in citrus leaves and inoculate OD 595 = 0.05-0.1 of citrus canker bacteria liquid, after adding bacteria, wait for it to dry, then put it into the incubator, the culture condition is dark condition, 20-25 ℃ culture for 24h; then a specific concentration of compound 67 is invaded into the filter paper and adhered to the leaves, DMSO is used as a positive control, TC is used as a negative control; then put it into the incubator, the culture condition is light condition, 10-20 ℃ culture for 24h; finally, the incubator condition is changed to a day-night switching mode (daytime 10-15h, 25 ℃, humidity greater than 80%; nighttime 10-15h, 20 ℃, humidity greater than 80%). The control efficiency calculation method is the same as above.

[0127] The examples of the present invention are used to illustrate the technical solutions of the present invention, but the contents of the examples are not limited thereto. The experimental results of the target compounds are shown in Table 8.

[0128] Table 8 Protective and therapeutic activities of the compounds of the present application against citrus canker bacteria

[0129]

[0130] As can be seen from Table 7, in the in vivo test, the target compound 67 at a concentration of 200 μg / mL showed good therapeutic activity (38.55%) and protective activity (56.46%) against citrus canker bacteria, which was better than the control drug thiophanate-methyl (therapeutic activity 21.63%; protective activity 26.72%), indicating that this series of compounds can be used to prepare pesticides against plant pathogenic bacteria.

Claims

1. An isopropanolamine compound containing α-phosphoramidate, or a salt, stereoisomer, or solvate thereof, characterized in that: The compound has a structure as shown in general formula (I): Wherein, R1, R2, R3, R4, and R5 are each independently selected from one or more of hydrogen, any substituted or unsubstituted alkyl, any substituted or unsubstituted alkoxy, any substituted or unsubstituted cycloalkyl, any substituted or unsubstituted aryl, and any substituted or unsubstituted heteroaryl.

2. The α-phosphoramidate compound containing isopropanolamine according to claim 1, or its salt, stereoisomer, solvate, characterized in that: R1 is selected from hydrogen, methyl, ethyl, propyl, phenyl, 1,3-dichlorophenyl, 1,4-dichlorophenyl, 1,3-difluorophenyl, 1,4-difluorophenyl, tert-butylphenyl, chlorophenyl, bromophenyl, iodophenyl, fluorophenyl, trifluorophenyl, methoxyphenyl; R2 and R3 are each independently selected from hydrogen, methyl, ethyl, propyl, butyl, cyclopropyl, -CH2CH2OH, phenyl, benzyl, R2 and R3 may be connected to each other to form a ring selected from R4, R5 are each independently selected from hydrogen, methyl, ethyl, and propyl.

3. The α-phosphoramidate compound containing isopropanolamine according to claim 1, or a salt, stereoisomer or solvate thereof, characterized in that Selected from the following compounds:

4. A method for preparing an α-phosphoramidate compound containing isopropanolamine according to any one of claims 1 to 3, or an intermediate of a salt, stereoisomer or solvate thereof, characterized in that As shown below: Wherein R1, R4 and R5 are as shown in any one of claims 1-3.

5. The method for preparing the isopropanolamine compound containing α-phosphoramidate according to any one of claims 1 to 3, or its salt, stereoisomer, or solvate, characterized in that The steps include: Preferably, the method further comprises the following steps: Most preferably, the method further comprises the following steps: Wherein R1, R2, R3, R4 and R5 are as described in claim 1.

6. A composition characterized in that Contains the compound according to any one of claims 1 to 3, or a salt, stereoisomer, solvate thereof, and an agriculturally usable adjuvant or fungicide, insecticide or herbicide; preferably, the composition is in the form of emulsifiable concentrate, dust, wettable powder, granule, aqueous solution, suspension, ultra-low volume spray, soluble powder, microcapsule, smoke agent, aqueous emulsion, water-dispersible granule.

7. Use of the compound according to any one of claims 1 to 3, or its salt, stereoisomer, solvate, or the composition according to claim 6 for preventing and controlling agricultural pests and diseases, preferably, the agricultural pests and diseases are viral diseases, plant bacterial or fungal diseases; more preferably, the agricultural pests and diseases are plant viruses, plant leaf blight and plant canker; most preferably, the agricultural pests and diseases are cucumber mosaic virus, tobacco mosaic virus, potato virus Y, rice bacterial leaf blight, cucumber bacterial leaf blight, konjac bacterial leaf blight, citrus canker, kiwi canker, grape canker, tomato canker, apple canker, rice bacterial leaf streak disease, cucumber gray mold, pepper wilt pathogen, rapeseed sclerotinia, wheat fusarium rust, potato late blight.

8. A method for preventing and controlling agricultural pests and diseases, characterized in that: The compound according to any one of claims 1 to 3, or its salt, stereoisomer, solvate, or the composition according to claim 6 is allowed to act on a harmful substance or its living environment; preferably, the agricultural pests and diseases are plant viruses, plant bacterial or fungal diseases; more preferably, the agricultural pests and diseases are cucumber mosaic virus, tobacco mosaic virus, potato virus Y, rice bacterial blight, tobacco bacterial wilt, cucumber bacterial blight, konjac bacterial blight, citrus canker, kiwi canker, grape canker, tomato canker, apple canker, rice bacterial leaf streak disease, cucumber gray mold, pepper wilt pathogen, rapeseed sclerotinia, wheat fusarium head blight, potato late blight.

9. A method for protecting plants from agricultural pests, comprising a method step in which the plant is contacted with a compound according to any one of claims 1 to 3, or a salt, stereoisomer, solvate thereof, or a composition according to claim 6.

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