Method for removing benzyl protecting group by using activated water and application of method in controllable release of medicine

Abstract

The invention relates to a method for removing a benzyl protecting group by using activated water and application of the method in controllable release of drugs. The method can be used for controllable release of salicylic acid (SA) and fenhexamid (FEN), enhancement of plant disease resistance and prevention and control of pathogenic fungi.

Description

Technical Field

[0001] The present invention belongs to the field of agricultural chemical synthesis methods, and specifically relates to a method for removing benzyl and benzyl derivatives by hydrogen transfer using rhodium and borane in activated water. At the same time, the method is applied to salicylic acid derivatives and fenhexamid derivatives containing benzyl and benzyl derivatives to enhance the resistance to salicylic acid immune activation and the pathogen control properties of fenhexamid. Background Art

[0002] Benzyl and benzyl derivatives are widely used protecting groups in synthetic chemistry, but the selective removal of such protecting groups remains a very challenging problem. Currently, the removal of such protecting groups usually requires the use of hydrogen / palladium on carbon and strong bases and strong reducing agents. The above methods are dangerous, have poor chemical selectivity, and are subject to high pressure in practical applications. There is an urgent need to develop new methods for removing benzyl protecting groups that are highly safe, highly selective, and environmentally friendly.

[0003] The search has not found any reports on the removal of benzyl and benzyl derivative protecting groups by chemically activating water. Based on this, we have developed a method of synergistically activating water with borane and rhodium using the tetracoordination characteristics of borane and the extremely high catalytic properties and low toxicity of rhodium to remove the benzyl group by hydrogen transfer. This method provides a green, safe, highly selective and biocompatible method for removing benzyl protecting groups.

[0004] At the same time, due to its excellent biocompatibility, the method is applied to immune activators and bactericides such as salicylic acid (SA) and fenhexamid (FEN), and its release rate in vivo and in vitro can be controlled extremely well, which will not affect plant growth, and can play or improve the effect of immune activation and control of pathogens. Among them, salicylic acid, as an endogenous hormone of plants, regulates its own immune disease resistance. However, since it will inhibit the growth of plants itself while activating immune disease resistance, it seriously affects its application in immune activators. Therefore, by controlling the release of salicylic acid content, while alleviating its growth inhibition, enhancing its immune activation characteristics, and extending the duration of effect have far-reaching effects. And fenhexamid has the advantages of not being easy to produce drug resistance and good systemic absorption, but due to its large field dosage and unstable efficacy, it is limited to its wide application. Therefore, it is of great significance to reduce the dosage and improve the efficacy through sustained release. Summary of the invention

[0005] The purpose of the present invention is to overcome the defects of the prior art and provide a green, highly biocompatible catalyst-induced method for selectively removing benzyl and / or benzyl derivative protecting groups. The method mainly completes water activation by coordination between the catalyst and water, followed by hydrogen atom transfer. After the substrate enters, the benzyl group is cleaved to generate a benzyl intermediate, and after another molecule of water is activated, a selectively deprotected target product is obtained. The second purpose of the present invention is to apply the method to the removal of salicylic acid derivatives and / or fenhexamid derivatives containing benzyl and benzyl derivatives, so as to achieve controlled release of salicylic acid and fenhexamid, and at the same time enhance the resistance to salicylic acid immune activation and the characteristics of fenhexamid in controlling pathogens.

[0006] In order to achieve the above object, the present invention provides a method for removing the benzyl protecting group by hydrogen transfer using a catalyst in coordination with activated water. The method has a chemical reaction formula shown in Formula I:

[0007]

[0008] Where: R 1 Selected from chain alkyl, cycloalkyl, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted heterocyclic aryl, acyl, acyloxy, ester, amide, carboxyl, carbonate, cyano, amino, substituted amino, preferably substituted and unsubstituted aryl, C1-C3 alkyl, amide, more preferably substituted and unsubstituted phenyl, C1-C3 alkyl, more preferably, the substitution refers to substitution by halogen, C1-C3 alkyl, C1-C3 alkyl substituted by halogen, carboxyl, amide or methoxy;

[0009] In addition, R 1 It can also be selected from substituted and unsubstituted glycine, alanine, valine, leucine, isoleucine, methionine (methionine), proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, wherein the substitution refers to substitution by halogen, C1-C3 alkyl, C1-C3 alkyl substituted by halogen, carboxyl, amide or methoxy;

[0010] X is selected from O, NR 2 or S preferably O or NR 2 ;

[0011] R 2Selected from hydrogen, chain alkyl, cycloalkyl, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, acyl, acyloxy, ester, amide, carbonate, cyano, sulfonyl, amino, substituted amino, preferably hydrogen, C1-C3 alkyl, substituted and unsubstituted phenyl, more preferably, the substitution refers to substitution by halogen, C1-C3 alkyl, C1-C3 alkyl substituted by halogen, carboxyl, amide or methoxy;

[0012] R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 are the same or different and are selected from hydrogen, alkyl, cycloalkyl, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, halogen, hydroxyl, sulfhydryl, nitro, cyano, carboxyl, ester, acyl, acyloxy, alkoxy, sulfonyl, amine, substituted amine, and amide;

[0013] Ar is selected from benzene, naphthalene, anthracene, phenanthrene, fluorene, pyridine, quinoline, isoquinoline, indole, pyrrole, pyrazine, pyrimidine, furan, benzofuran, benzothiazole, benzothiophene, and carbazole.

[0014] Preferably, the R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 is selected from hydrogen or halogen, wherein the halogen is any one of fluorine, chlorine and bromine.

[0015] Preferably, Ar is benzene.

[0016] The method of the present invention specifically comprises the following steps: dissolving a substrate protected by a benzyl or benzyl derivative protecting group in a solvent, adding a catalyst and an aqueous solution, mixing evenly, and reacting the mixed solution at 25-150° C.; after the reaction is complete, adding water to quench, extracting with ethyl acetate, collecting the ethyl acetate phase, washing with water and a saturated sodium chloride aqueous solution in sequence, drying with anhydrous sodium sulfate, and concentrating by column chromatography to obtain a selectively deprotected target product.

[0017] In addition, in order to achieve the above-mentioned purpose, the present invention provides a method for removing salicylic acid derivatives and fenhexamid derivatives containing benzyl and benzyl derivatives, as shown in Formula II and Formula III,

[0018]

[0019] Wherein, Q is selected from Q1 or Q2:

[0020]

[0021] Specifically, R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 are the same or different and are selected from hydrogen, alkyl, cycloalkyl, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, halogen, hydroxyl, thiol, nitro, cyano, carboxyl, ester, acyl, acyloxy, alkoxy, sulfonyl, amino, substituted amino and amide; Ar is selected from benzene, naphthalene, anthracene, phenanthrene, fluorene, pyridine, quinoline, isoquinoline, indole, pyrrole, pyrazine, pyrimidine, furan, benzofuran, benzothiazole, benzothiophene and carbazole.

[0022] Preferably, the R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 is selected from hydrogen or halogen, wherein the halogen is any one of fluorine, chlorine and bromine.

[0023] Preferably, Ar is selected from benzene.

[0024] Specifically, the catalyst includes a first catalyst and a second catalyst.

[0025] Preferably, the first catalyst is selected from boranes such as tetrakis dimethylaminodiborane, tetrahydroxylated diborane, biboronic acid pinacol ester, bis-catechol borate, bis-(2,4-dimethyl-2,4-pentanediol) borate, bis-(2-methyl-2,4-pentanediol) borate, 9-boranebicyclo[3.3.1]nonane dimer, and tetrakis(pyrrolidino)diborane.

[0026] More preferably, the first catalyst is selected from one or both of tetrakis dimethylaminodiborane and / or tetrahydroxylated diboron.

[0027] Preferably, the second catalyst is selected from (1,5-cyclooctadiene)chlororhodium (I) dimer, methoxy (cyclooctadiene) rhodium (I) dimer, dimerized hydroxy (1,5-cyclooctadiene) rhodium (I), dimerized rhodium acetate, triphenylphosphine rhodium chloride, bis (1,5-cyclooctadiene) rhodium hexafluorophosphate, chlorobis (ethylene) rhodium (I) dimer, acetyl naphthyl bis (cyclooctene) rhodium (I), chloronorbornadiene rhodium dimer, dicyclooctene rhodium chloride dimer, tetrakis (triphenylphosphine) rhodium hydride (I), bis (1,5-cyclooctadiene) rhodium tetrafluoroborate (I), (1,5-hexadiene) rhodium chloride (I) dimer, diphenylphosphine (1,5-cyclooctadiene) rhodium (I) dimer, and the like.

[0028] More preferably, the second catalyst is selected from one or both of (1,5-cyclooctadiene)chlororhodium (I) dimer and / or dipolymerized hydroxy(1,5-cyclooctadiene)rhodium (I).

[0029] Preferably, the first catalyst accounts for 50 mol%-500 mol%, preferably, the first catalyst accounts for 200 mol%; the second catalyst accounts for 0.5 mol%-20 mol%, preferably, the second catalyst accounts for 5 mol%. The above-mentioned proportions are all molar proportions with the starting raw materials.

[0030] Preferably, the solvent is selected from one or more of n-hexane, cyclohexane, heptane, benzene, toluene, xylene, trimethylbenzene, trifluorotoluene, chlorobenzene, carbon tetrachloride, chloroform, dichloromethane, 1,2-dichloroethane, tetrahydrofuran, ether, carbon disulfide, chloropropane, ethyl bromide, isopropyl ether, nitromethane, butyl acetate, n-pentane, dioxane, methyl acetate, methyl tert-butyl ether, petroleum ether, acetone, acetonitrile, methanol, ethanol, isopropanol, n-propanol, n-butanol, isobutanol, tert-butanol, water, pyridine, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, ethyl acetate, trifluoroethanol, and hexafluoroisopropanol.

[0031] Preferably, the solvent is selected from one or both of tetrahydrofuran and / or methanol.

[0032] Preferably, the volume ratio of water and solvent can be selected from 0.1%-100%. Preferably, the volume ratio of water and solvent is 50%.

[0033] The reaction conditions include a reaction temperature selected from 20-150°C, preferably 25-150°C, and a reaction time of 0.2-72h.

[0034] Preferably, the reaction temperature is selected from 25-37°C, and the reaction time is 1-6h, preferably 2-5h, such as 2h, 3h, 4h, 5h, etc.

[0035] In a third aspect, the present invention seeks to protect the use of the aforementioned method in the controlled release of salicylic acid and fenhexamid.

[0036] In a fourth aspect, the present invention claims the application of the aforementioned method in the following (A1) or (A2):

[0037] (A1) Application in enhancing plant resistance to disease and insects or inhibiting pathogens;

[0038] (A2) Use in the preparation of pesticide agents for enhancing plant resistance to disease and insects or inhibiting pathogenic bacteria;

[0039] Preferably, the application comprises the following steps: preparing a first catalyst and a second catalyst in any molar ratio of 25 mol:1 mol to 200 mol:1 mol into a fresh mixture and spraying the mixture on plants pretreated with 0.25 mmol-5 mmol SA-1 and / or FEN-1; preferably, the first catalyst is tetrakis dimethylaminodiborane, the second catalyst is (1,5-cyclooctadiene)chlororhodium (I) dimer, and the ratio of the first catalyst to the second catalyst is 40 mol:1 mol;

[0040] Preferably, the configuration of the fresh mixture of the first catalyst and the second catalyst contains 0.1%-3% of a reaction solvent, and the reaction solvent is selected from one or more of n-hexane, cyclohexane, heptane, benzene, toluene, xylene, trimethylbenzene, trifluorotoluene, chlorobenzene, carbon tetrachloride, chloroform, dichloromethane, 1,2-dichloroethane, tetrahydrofuran, ether, carbon disulfide, chloropropane, ethyl bromide, isopropyl ether, nitromethane, butyl acetate, n-pentane, dioxane, methyl acetate, methyl tert-butyl ether, petroleum ether, acetone, acetonitrile, methanol, ethanol, isopropanol, n-propanol, n-butanol, isobutanol, tert-butanol, water, pyridine, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, ethyl acetate, trifluoroethanol, and hexafluoroisopropanol; preferably, the solvent is selected from one or both of tetrahydrofuran or methanol.

[0041] In a fifth aspect, the present invention also protects the use of agents containing the SA-1 and / or FEN-1 described above in immune activators, plant growth regulators and fungicides.

[0042] In a sixth aspect, the present invention also protects the use of a reagent containing the first catalyst and the second catalyst described above in the preparation of agricultural adjuvants and agricultural plant growth regulators.

[0043] Beneficial Effects

[0044] The above method of the present invention can remove benzyl and benzyl derivative protecting groups in a green, efficient, safe, selective and biocompatible manner. The method can be applied to the removal of salicylic acid derivatives containing benzyl and benzyl derivative protecting groups, and can controllably release salicylic acid and fenhexamid, thereby enhancing plant disease resistance and controlling pathogen infection. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 It is the liquid chromatogram of the reaction system of compound 46, 46a, 46. The abscissa is the peak time (min), and the ordinate is the peak height (mv).

[0046] Figure 2 The resistance of Nicotiana benthamiana leaves to pepper phytophthora after treatment with the agent; Figure 2 Figure a: Disease resistance results of Nicotiana benthamiana leaves treated with B-Rh, Bn-SA, Bn-SA+B-Rh, and SA against pepper phytophthora; Figure 2 Figure b: Diameter (mm) of lesion area on Nicotiana benthamiana leaves after treatment with various drugs.

[0047] Figure 3 The resistance of Nicotiana benthamiana plants to pepper phytophthora after treatment with the agent; Figure 3 Figure a: Disease resistance results of Nicotiana benthamiana plants treated with B-Rh, Bn-SA, Bn-SA+B-Rh, and SA against spores of Phytophthora capsici; Figure 3 Figure b: Relative control effect (%) of each drug on Nicotiana benthamiana plants.

[0048] Figure 4 The resistance of Nicotiana benthamiana plants to pepper phytophthora after being treated with the agent for different days; Figure 4 Figure a: Disease resistance results of Nicotiana benthamiana plants to pepper phytophthora spores after being treated with Bn-SA+B-Rh and SA+B-Rh for different days; Figure 4 Figure b: Relative control effect (%) of Bn-SA+B-Rh and SA+B-Rh on Nicotiana benthamiana plants after treatment on different days.

[0049] Figure 5 This is the control effect of rapeseed sclerotinia after treating rapeseed leaves with different concentrations of pesticides. DETAILED DESCRIPTION

[0050] The specific embodiments of the present invention are described in detail below. It should be understood that the specific embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0051] The endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0052] The reagents and methods used in the examples, unless otherwise specified, all use conventional reagents and conventional methods. In the reactions involved in the present invention, various post-treatment methods conventionally used in the art can be used to post-treat the obtained products. The post-treatment methods are not limited to: extraction, recrystallization, washing, drying, filtration, column chromatography, etc. The present invention will not be repeated here, and the post-treatment methods involved in the examples are only used for illustrative purposes, and do not mean that they are operations that must be adopted. Those skilled in the art can use other conventional means to replace them.

[0053] Example 1

[0054] The removal reaction of compound 29 containing a benzyl derivative protecting group is as follows:

[0055]

[0056] The raw material 29 (1 mmol) was dissolved in 1.5 mL of tetrahydrofuran (THF), and 0.05 mmol of (1,5-cyclooctadiene) rhodium chloride (I) dimer ([RhCl(cod)2]) and 2 mmol of tetramethylaminodiborane (B2(NMe2)4) were weighed and mixed evenly, and 1.5 mL of water was added. After sealing with a rubber stopper, the mixture was reacted at room temperature for 2 h, and the reaction was monitored by TLC until the end. After adding water to quench the reaction, the tetrahydrofuran was removed by vacuum concentration, and ethyl acetate (15 mL×3) was added to extract the reaction three times. The organic phases were combined and washed three times with saturated brine (50 mL×3). The organic phases were dried over anhydrous sodium sulfate, concentrated in vacuo, and column chromatography to obtain a white solid 29a with a yield of 91%. 1 HNMR(500MHz,Chloroform-d)δ6.83–6.75(m,4H),4.86(s,1H),3.76(s,3H).

[0057] Example 2

[0058] The removal reaction of compound 25 containing a benzyl protecting group, the chemical reaction formula is as follows:

[0059]

[0060] The raw material 25 (1 mmol) was dissolved in 1.5 mL of tetrahydrofuran (THF), and 0.1 mmol of (1,5-cyclooctadiene)chlororhodium (I) dimer and 2 mmol of tetramethylaminodiborane were weighed and mixed well. Then, 2 mmol of triethylamine and 1.5 mL of water were added respectively. After sealing with a rubber stopper, the mixture was reacted at room temperature for 5 h. The reaction was monitored by TLC until completion. After adding water to quench the reaction, the tetrahydrofuran was removed by vacuum concentration, and ethyl acetate (15 mL × 3) was added to extract the reaction three times. The organic phases were combined, washed twice with a saturated sodium bicarbonate solution (50 mL × 2), and then washed twice with a saturated saline solution (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, concentrated in vacuo, and column chromatography to obtain a light yellow solid 25a with a yield of 94%. 1 H NMR(500MHz,Chloroform-d)δ7.18(t,J=7.0Hz,2H),6.70(t,J=7.5Hz,1H),6.62(d,J =1.0Hz,1H),6.61(s,1H),3.60(s,1H),3.15(q,J=7.0Hz,2H),1.25(t,J=7.0Hz,3H).

[0061] Example 3

[0062]

[0063] The raw material 46 (1 mmol), (1,5-cyclooctadiene)chlororhodium (I) dimer (0.05 mmol) and tetrakis dimethylaminodiborane (2 mmol) were weighed and mixed evenly, and 3 mL of water was added. After sealing with a rubber stopper, the mixture was reacted at room temperature for 2 h, and the reaction was monitored by HPLC until completion.

[0064] Liquid phase analysis was performed on an Agilent 1240 liquid phase (Agilent Technologies, USA) using an Eclipse XDB-C18 column (5 μm, 4.6 × 250 mm) with a precolumn. The analysis conditions for 46 and 46a were 20-100% methanol: ddH2O, linear gradient for 20 min, followed by 100% methanol: ddH2O for 8 min, and finally 20% methanol: ddH2O for 5 min, with a flow rate of 1 mL / min. Figure 1 As shown, the relative yield calculated based on the peak area was 99%.

[0065] Example 4

[0066]

[0067] The raw material Bn-SA (1mmol) was dissolved in 1.5mL tetrahydrofuran (THF), and 0.05mmol of (1,5-cyclooctadiene)chlororhodium (I) dimer and 2mmol of tetramethylaminodiborane were weighed and mixed evenly, and 1.5mL of water was added. After sealing with a rubber stopper, the reaction was allowed to react at room temperature for 2h, and the reaction was monitored by TLC until the end. After adding water to quench the reaction, the tetrahydrofuran was removed by vacuum concentration, and ethyl acetate (15mL×3) was added to extract the reaction three times. The organic phases were combined and washed three times with saturated brine (50mL×3), and the organic phases were dried over anhydrous sodium sulfate, concentrated in vacuo, and column chromatography to obtain white solid SA with a yield of 97%. 1 HNMR(500MHz,Chloroform-d)δ10.40(s,1H),7.94(d,J=8.0Hz,1H),7.54(t,J=7.0Hz,1H),7.02(d,J=8.5Hz,1H),6.95(t,J=7.5Hz,1H).

[0068] Example 5

[0069]

[0070] The raw material Bn-FEN (1mmol) was dissolved in 1.5mL tetrahydrofuran (THF), and 0.05mmol of (1,5-cyclooctadiene)chlororhodium (I) dimer and 2mmol of tetramethylaminodiborane were weighed and mixed evenly, and 1.5mL of water was added. After sealing with a rubber stopper, the mixture was reacted at room temperature for 2h, and the reaction was monitored by TLC until the end. After adding water to quench the reaction, the tetrahydrofuran was removed by vacuum concentration, and ethyl acetate (15mL×3) was added to extract the reaction three times. The organic phases were combined and washed three times with saturated brine (50mL×3), and the organic phases were dried over anhydrous sodium sulfate, concentrated in vacuum, and column chromatography to obtain white solid FEN with a yield of 97%. 1 H NMR(500MHz,Chloroform-d)δ8.14(d,J=9.0Hz,1H),7.77(s,1H),6.97(d,J=9.5Hz,1H) ,5.64(s,1H),2.04(t,J=6.5Hz,2H),1.66–1.61(m,2H),1.56–1.36(m,6H),1.28(s,3H).

[0071] Example 6

[0072] Nicotiana benthamiana seeds were sown in a seedling medium (nutrient soil: vermiculite = 2:1), cultured for 7 days under the conditions of light:darkness = 16h:8h and a temperature of 25±1°C, and then transplanted into a square box with a diameter of 6 cm containing culture soil (nutrient soil: vermiculite = 3:1), and cultured for another 4 weeks under the above conditions before use.

[0073] N. benthamiana plants were treated with 1 mL Bn-SA (1 mM) for 24 h and then sprayed with B-Rh complex (0.025 mM [RhCl (cod) 2] and 1 mM B2 (NMe2) 4). The control group was treated with 1 mL H2O, SA (1 mM), Bn-SA (1 mM) and B-Rh complex (0.025 mM [RhCl (cod) 2] and 1 mM B2 (NMe2) 4) respectively. After 24 h, leaves of uniform size were cut and inoculated with pepper Phytophthora blocks with a diameter of 0.5 mm. After culturing pepper Phytophthora LT263 on V8 plates at 25 ± 1 ° C in the dark for 3-4 days, the outermost mycelial block of the colony was picked. Water treatment was used as a blank control. After inoculation, it was cultured at 25 ° C in the dark for 36 h. The plaque diameter was measured using the cross method, and the initial diameter of the pepper Phytophthora block of 0.5 mm was deducted from the final data.

[0074] like Figure 2 As shown in the figure, the lesion area after treatment with B-Rh (15.4 mm) and Bn-SA (16.03 mm) was basically the same as that after treatment with H2O (16.67 mm), but the joint use of Bn-SA + B-Rh restored the disease resistance of the leaves, with a lesion area of ​​2.03 mm, which was smaller than 3.39 mm after treatment with SA alone. The above results all indicate that the use of B Rh promotes the removal of benzyl in Bn-SA, releases SA, and induces enhanced disease resistance in the leaves, which is better than the disease resistance after SA alone.

[0075] Example 7

[0076] The cultivation method of Nicotiana benthamiana is the same as that in Example 6.

[0077] Method for inducing spores of pepper phytophthora: After culturing pepper phytophthora LT263 on a V8 plate at 25±1℃ in the dark for 3-4 days, pick the outermost mycelium block of the colony and put it into a culture dish containing 15mL V8 liquid culture medium. Cultivate in the dark at 25±1℃ for 3 days, discard the culture solution and wash the mycelium repeatedly with sterilized tap water three times, and then culture in the dark at 25℃ for 24h. Transfer the culture dish to 4℃ for 30min to stimulate spore production by low temperature. After 30min, return it to 25±1℃ for 30min, place the culture dish under a microscope, and observe the mycelium and whether it produces spores. After successful spore production, a platelet counter can be used to count and prepare the corresponding concentration of zoospore liquid according to experimental needs.

[0078] V8 solid culture medium, each liter of culture medium includes: 100mL V8 fruit and vegetable juice, 1g CaCO3, 20g agar, and 900ml water.

[0079] V8 liquid culture medium, each liter of culture medium includes: 100mL V8 fruit and vegetable juice, 1g CaCO3, 900ml water. When preparing the liquid culture medium, mix the V8 fruit and vegetable juice with calcium carbonate, centrifuge at 8000rpm for 5min, discard the precipitate, keep the supernatant, and add water to make up to 1L.

[0080] Weigh the above weight ratio, make up to volume with ultrapure water, dispense into Erlenmeyer flasks, sterilize at 121℃ for 25min, and prepare the required culture medium.

[0081] Treat N. benthamiana plants with 1 mL Bn-SA (1 mM) for 24 h, then spray with B-Rh complex (0.025 mM [RhCl (cod) 2] and 1 mM B2 (NMe2) 4). The control group was treated with 1 mL H2O, SA (1 mM), Bn-SA (1 mM) and B-Rh complex (0.025 mM [RhCl (cod) 2] and 1 mM B2 (NMe2) 4) respectively. The blank control was sprayed with water. After 24 h, the plants were sprayed with zoospore liquid. After spraying, the plants were placed in a moisturizing environment for 4 days, and the disease index was counted and photographed when the plants became sick.

[0082] The results are as follows Figure 3As shown, the control effect of B-Rh (14.81%) and Bn-SA (9.26%) after drug treatment is basically the same as that after H2O treatment. The plants wilt, and the leaves have water-soaked spots and curl. This shows that the use of B-Rh and Bn-SA alone cannot cause disease resistance in plants. After treatment with 1mM SA, the plants remained upright, without wilting, and the leaves did not curl. The disease resistance effect was excellent (91.67%). However, the joint use of Bn-SA + B-Rh enhanced the disease resistance of the plants, and the disease resistance effect reached 94.44%, which is better than the disease resistance effect of SA alone. The above results show that Bn-SA + B-Rh can be used to activate the disease resistance of living plants, and its disease resistance effect is better than the use of SA alone.

[0083] Example 8

[0084] The cultivation method of Nicotiana benthamiana is the same as that in Example 6.

[0085] The method for inducing spores of Phytophthora capsici is the same as that in Example 7.

[0086] SA (1mM), Bn-SA (1mM) and Bn-SA (3mM) solutions were used for treatment respectively. After 24h, B-Rh complex (0.005mM [RhCl (cod)]2 and 0.5mM B2 (NMe2)4) was sprayed on Nicotiana benthamiana plants. The same procedure was repeated every other day on new Nicotiana benthamiana plants for a total of 10 times. The cultivation was continued in a light incubator with a cycle of 16 hours of light (25℃) / 8 hours of darkness (25℃). After the 10th group was sprayed with B-Rh complex, 1mL of pepper spore solution (1*10^5 spores / mL) was sprayed on all treated plants at the same time at an interval of 24h, and 1mL was maintained for each plant. After spraying, the plants were placed in a moisturizing environment for 4d, and the disease index was counted and photographed when the plants became sick. Each group was repeated three times.

[0087] The results are as follows Figure 4 As shown, in order to exclude the effect of B-Rh on the persistence period, B-Rh was also added to the SA treatment as a control. On the 9th day after the 1mM SA+B-Rh treatment, the disease resistance of the plants disappeared. Correspondingly, the disease resistance of the Nicotiana benthamiana plants treated with 1mM Bn-SA+B-Rh disappeared on the 13th day. Relative to the SA persistence period, the co-use of Bn-SA and B-Rh extended the persistence period by 4 days. The disease resistance persistence period of Nicotiana benthamiana plants treated with 3mM Bn-SA+B-Rh was extended by 8 days. In short, the co-use of Bn-SA+B-Rh can not only restore the disease resistance of the plants, but also occasionally extend the disease resistance persistence period of the plants.

[0088] Example 9

[0089] The petioles of freshly picked rapeseed leaves (from the Baima Experimental Base of Nanjing Agricultural University, susceptible) were treated with water for 24 hours. Spray 1mL Bn-FEN, and 24 hours later, spray 1mL B-Rh complex (0.025mM [RhCl (cod) 2] and 1mM B2 (NMe2) 4), and inoculate 0.5mm outermost circle of rapeseed Sclerotinia sclerotiorum at symmetrical positions of the leaves (avoiding the veins). Cultured in the dark at 25℃ for 2 days, the diameter of the lesions (mm) was measured by the cross method. Meizu treatment was repeated 3 times. Rapeseed Sclerotinia sclerotiorum was cultured on PDA medium for 2 days.

[0090] PDA medium: 200 g potato, 20 g glucose, 20 g agar, and make up to 1 L with water.

[0091] In order to test whether the control effect of Bn-FEN+B-Rh on pathogens is better than that of FEN alone, we conducted an experiment on rapeseed leaves to control rapeseed sclerotinia. Figure 5 As shown in Table 1, B-Rh and 1 mg / mL Bn-FEN alone cannot prevent the occurrence of rapeseed sclerotinia, and the lesion area is basically the same as that of the control group. However, the combined use of Bn-FEN + B-Rh restored the control of rapeseed leaves against rapeseed sclerotinia, and its control effect was improved compared with the use of FEN alone. It can be found from Table 1 that the combined use of 1 mg / mL, 0.50 mg / mL, 0.25 mg / mL Bn-FEN and B-Rh can improve their control effect, and the improvement is close to 20%. In short, the combined use of Bn-FEN + B-Rh can not only restore the control effect of plants against pathogens, but also improve the control effect to a certain extent compared with FEN.

[0092] Table 1 Relative control effect of rapeseed leaves treated with different concentrations of pesticides on rapeseed sclerotinia (%)

[0093]

[0094] In conclusion, it can be seen from the above results that the above method of the present invention can not only selectively remove benzyl and benzyl derivatives, but also has the advantages of being green, environmentally friendly, highly safe, and highly biocompatible. At the same time, applying this method to known immunohormones or fungicides can significantly improve their efficacy and prolong the duration of the drug, which provides great help in the subsequent drug synthesis and the development of agricultural drug adjuvants.

[0095] The preferred embodiments of the present invention are described in detail above, but within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention. The various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further explain various possible combinations.

Claims

1. A method for removing benzyl and benzyl derivatives, characterized in that: The removal method has a reaction route shown in Formula I, which is as follows: water activation is completed after the catalyst and water are coordinated, followed by hydrogen atom transfer, and after the substrate enters, the benzyl group is cleaved to generate a benzyl intermediate, which is then activated by another molecule of water to obtain a selectively deprotected target product; Where: R 1 is selected from the group consisting of alkyl, cycloalkyl, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, acyl, acyloxy, ester, amide, carboxyl, carbonate, cyano, amino, substituted amino, preferably substituted and unsubstituted aryl, C1-C3 alkyl, amide, more preferably substituted and unsubstituted phenyl, C1-C3 alkyl, and / or, R 1 Selected from substituted and unsubstituted glycine, alanine, valine, leucine, isoleucine, methionine (methionine), proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, wherein the substitution refers to substitution by halogen, C1-C3 alkyl, C1-C3 alkyl substituted by halogen, carboxyl, amide or methoxy; X is selected from O, NR 2 or S is preferably O or NR 2 ; R 2 Selected from hydrogen, alkanyl, cycloalkyl, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, acyl, acyloxy, ester, amide, carbonate, cyano, sulfonyl, amino, substituted amino, preferably hydrogen, C1-C3 alkyl, substituted and unsubstituted phenyl; R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 are the same or different and are selected from hydrogen, alkyl, cycloalkyl, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, halogen, hydroxyl, sulfhydryl, nitro, cyano, carboxyl, ester, acyl, acyloxy, alkoxy, sulfonyl, amine, substituted amine, and amide; Ar is selected from benzene, naphthalene, anthracene, phenanthrene, fluorene, pyridine, quinoline, isoquinoline, indole, pyrrole, pyrazine, pyrimidine, furan, benzofuran, benzothiazole, benzothiophene, and carbazole, and is preferably benzene.

2. The method according to claim 1, characterized in that: The specific steps are as follows: (1) dissolving a substrate protected by a benzyl or benzyl derivative protecting group in a reaction solvent, and reacting in the presence of a catalyst and an aqueous solution; (2) After the reaction in step (1) is completed, water is added to quench the reaction and then the mixture is extracted with ethyl acetate. The ethyl acetate phase is collected, washed, dried, concentrated, and subjected to column chromatography to obtain the target product with selective deprotection.

3. A method for removing the protecting group of benzyl or benzyl derivative in a salicylic acid derivative or fenhexamid derivative containing benzyl or benzyl derivative, characterized in that: The reaction route of the method is shown in Formula II or Formula III: Wherein, Q is selected from Q1 or Q2; Among them, R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 are the same or different and are selected from hydrogen, alkyl, cycloalkyl, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, halogen, hydroxyl, sulfhydryl, nitro, cyano, carboxyl, ester, acyl, acyloxy, alkoxy, sulfonyl, amine, substituted amine, and amide; Ar is selected from benzene, naphthalene, anthracene, phenanthrene, fluorene, pyridine, quinoline, isoquinoline, indole, pyrrole, pyrazine, pyrimidine, furan, benzofuran, benzothiazole, benzothiophene, and carbazole; The specific steps of the method are as follows: (1) dissolving a substrate salicylic acid derivative or fenhexamid derivative protected by a benzyl or benzyl derivative protecting group in a reaction solvent, and reacting in the presence of a catalyst and an aqueous solution; (2) After the reaction in step (1) is completed, water is added to quench the reaction and then the mixture is extracted with ethyl acetate. The ethyl acetate phase is collected, washed, dried, concentrated, and subjected to column chromatography to obtain the target product with selective deprotection.

4. The method according to claims 1-3, characterized in that: The catalyst is composed of a first catalyst and a second catalyst; the first catalyst is selected from one or more of tetrakis dimethylaminodiborane, tetrahydroxylated diborane, biboric acid pinacol ester, bis-catechol borate, bis-(2,4-dimethyl-2,4-pentanediol) borate, bis-(2-methyl-2,4-pentanediol) borate, 9-boronobicyclo[3.3.1]nonane dimer, and tetrakis(pyrrolidino)diborane; the second catalyst is selected from one or more of (1,5-cyclooctadiene)chlororhodium(I) dimer, methoxy(cyclooctadiene)rhodium(I) dimer, One or more of diisocyanate, ... Preferably, the first catalyst is selected from one or both of tetrakis dimethylaminodiborane and / or tetrahydroxylated diboron, and the second catalyst is selected from one or both of (1,5-cyclooctadiene)chlororhodium (I) dimer and / or dipolymerized hydroxy(1,5-cyclooctadiene)rhodium (I); Preferably, the proportion of the first catalyst is selected from 50mol%-500mol%, preferably, the proportion of the first catalyst is 200mol%; the proportion of the second catalyst is selected from 0.5mol%-20mol%, preferably, the proportion of the second catalyst is 5mol%, and the above-mentioned proportions are all molar proportions with the content of the starting raw material.

5. The method according to claims 1-3, characterized in that: The solvent is selected from one or more of n-hexane, cyclohexane, heptane, benzene, toluene, xylene, trimethylbenzene, trifluorotoluene, chlorobenzene, carbon tetrachloride, chloroform, dichloromethane, 1,2-dichloroethane, tetrahydrofuran, ether, carbon disulfide, chloropropane, ethyl bromide, isopropyl ether, nitromethane, butyl acetate, n-pentane, dioxane, methyl acetate, methyl tert-butyl ether, petroleum ether, acetone, acetonitrile, methanol, ethanol, isopropanol, n-propanol, n-butanol, isobutanol, tert-butanol, water, pyridine, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, ethyl acetate, trifluoroethanol, and hexafluoroisopropanol; preferably, the solvent is selected from one or two of tetrahydrofuran and / or methanol. Preferably, the volume ratio of water and solvent is selected from 0.1%-100%; preferably, the volume ratio of water and solvent is 50%.

6. The method according to claims 1-3, characterized in that: The reaction conditions are as follows: the reaction temperature is selected from 25-150° C., and the reaction time is 0.2-72 h; preferably, the reaction temperature is selected from 25-37° C., and the reaction time is 1-6 h.

7. Use of the method according to any one of claims 1 to 6 in the controlled release of salicylic acid and / or fenhexamid.

8. Use of the method according to any one of claims 1 to 6 in the following (A1) or (A2): (A1) Application in enhancing plant resistance to disease and insects or inhibiting pathogens; (A2) Use in the preparation of pesticide agents for enhancing plant resistance to disease and insects or inhibiting pathogenic bacteria; Preferably, the application comprises the following steps: preparing a first catalyst and a second catalyst in any molar ratio of 25 mol:1 mol to 200 mol:1 mol into a fresh mixture and spraying the mixture on plants pretreated with 0.25 mmol-5 mmol SA-1 and / or FEN-1; preferably, the first catalyst is tetrakis dimethylaminodiborane, the second catalyst is (1,5-cyclooctadiene)chlororhodium (I) dimer, and the ratio of the first catalyst to the second catalyst is 40 mol:1 mol; Preferably, the configuration of the fresh mixture of the first catalyst and the second catalyst contains 0.1%-3% of a reaction solvent, and the reaction solvent is selected from one or more of n-hexane, cyclohexane, heptane, benzene, toluene, xylene, trimethylbenzene, trifluorotoluene, chlorobenzene, carbon tetrachloride, chloroform, dichloromethane, 1,2-dichloroethane, tetrahydrofuran, ether, carbon disulfide, chloropropane, ethyl bromide, isopropyl ether, nitromethane, butyl acetate, n-pentane, dioxane, methyl acetate, methyl tert-butyl ether, petroleum ether, acetone, acetonitrile, methanol, ethanol, isopropanol, n-propanol, n-butanol, isobutanol, tert-butanol, water, pyridine, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, ethyl acetate, trifluoroethanol, and hexafluoroisopropanol; preferably, the solvent is selected from one or both of tetrahydrofuran or methanol.

9. Use of the reagent containing SA-1 and / or FEN-1 according to claim 3 in immune activators, plant growth regulators and fungicides.

10. Use of a reagent containing the first catalyst and the second catalyst according to claim 4 in agricultural adjuvants and agricultural plant growth regulators.

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