Berberine derivative, preparation method and application thereof

Abstract

The application discloses a new berberine derivative, a preparation method and application thereof, and a structure formula is shown as follows: The berberine derivative has good bacteriostatic effect, and the bacteriostatic effect on part of bacteria is obviously better than that of berberine itself, and can be applied in agricultural prevention and treatment of crop diseases in a large amount.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of agrochemical technology, in particular to a novel berberine derivative and a preparation method and application thereof. BACKGROUND

[0002] Chemical control is the main technical means for crop disease control, but large-scale application will lead to increased residues and serious environmental pollution. It not only kills pests, but also kills natural enemies, thereby destroying the ecological balance. However, biological pesticides have high selectivity, are relatively safe to humans and animals, have little effect on non-target organisms, have low or no toxicity, and have little impact on the ecological environment. Therefore, developing various new biological fungicidal compounds is a solution to the problems faced by the pesticide industry.

[0003] Berberine, also known as coptis, is widely distributed in the plant kingdom and is the main alkaloid active ingredient of traditional heat-clearing and detoxifying Chinese medicine coptis. Studies have found that berberine has strong inhibitory activity on a variety of gram-positive bacteria, gram-negative bacteria and fungi in vitro and is widely used in clinical practice. In recent years, due to the green and sustainable development of ecology, the wide application of biological pesticides is advocated, and berberine-like compounds have been widely used in the field of crop disease control. Existing studies have found that berberine has strong fungicidal activity, and in the study of the fungicidal mechanism, it is found that berberine has a significant effect on the permeability of pathogenic fungal cell membranes, reduces the content of soluble proteins, and weakens the activity of intracellular succinate dehydrogenase, thereby blocking the absorption of nutrients.

[0004] By modifying the structure of berberine, novel and better fungicidal activity derivatives are obtained. Therefore, the present application is proposed. SUMMARY

[0005] The present application provides a novel berberine derivative, which is a novel compound. The fungicidal effect of the novel berberine derivative is better than that of berberine itself, and the fungicidal effect of some of the novel berberine derivatives is comparable to or even better than that of chemical pesticides. Therefore, the novel berberine derivative has good fungicidal potential.

[0006] The present application also provides a preparation method of the novel berberine derivative, which can obtain a high yield of the novel berberine derivative.

[0007] The present application further provides an application of the novel berberine derivative, in particular, an application in fungistatic and fungicidal aspects.

[0008] The present application is realized by the following technical scheme:

[0009] A novel berberine derivative, characterized in that the structure is as shown in the following formula:

[0010]

[0011] wherein the R is CH3O-, which is named p-methoxyacetophenone berberine, and the structural formula is as shown below:

[0012]

[0013] wherein the R is -NO2, which is named p-nitroacetophenone berberine, and the structural formula is as shown below:

[0014]

[0015] wherein the R is -H, which is named unsubstituted acetophenone berberine, and the structural formula is as shown below:

[0016]

[0017] wherein the R is -F, which is named p-fluoroacetophenone berberine, and the structural formula is as shown below:

[0018]

[0019] wherein the R is -Cl, which is named p-chloroacetophenone berberine, and the structural formula is as shown below:

[0020]

[0021] wherein the R is -OH, which is named p-hydroxyacetophenone berberine, and the structural formula is as shown below:

[0022]

[0023] The hydrogen spectrum data of the p-methoxyacetophenone berberine is as follows: 1 H NMR (400 MHz, DMSO) δ 10.01 (s, 1H), 8.97 (s, 1H), 8.43-8.38 (m, 1H), 8.27-8.23 (m, 1H), 8.19 (d, J = 9.2 Hz, 1H), 7.99 (t, J = 10.0 Hz, 1H), 7.82 (s, 1H), 7.12 (s, 1H), 6.19 (s, 1H), 5.92 (s, 1H), 4.96 (t, J = 6.3 Hz, 1H), 3.95 (s, 1H), 3.27-3.19 (m, 1H);

[0024] The carbon spectrum data is as follows: 13C NMR (101 MHz, DMSO) δ 194.39 (s), 150.78 (s), 150.39 (s), 149.57 (s), 148.21 (s), 146.21 (s), 142.33 (s), 139.34 (s), 138.11 (s), 133.55 (s), 131.20 (s), 129.87 (s), 127.26 (s), 124.48 (s), 123.75 (s), 121.53 (s), 120.92 (s), 120.62 (s), 108.94 (s), 105.97 (s), 102.59 (s), 75.76 (s), 57.64 (s), 55.95 (s), 26.88 (s).

[0025] The hydrogen spectrum data for p-nitroacetophenone berberine is: 1 H NMR (400 MHz, DMSO) δ 10.02 (s, 1H), 8.96 (s, 1H), 8.18 (d, J = 9.2 Hz, 1H), 7.98 (d, J = 8.7 Hz, 3H), 7.82 (s, 1H), 7.11 (s, 2H), 7.09 (s, 1H), 6.19 (s, 2H), 5.85 (s, 2H), 3.96 (s, 3H), 3.87 (s, 3H), 3.23 (t, J = 6.2 Hz, 2H);

[0026] The carbon spectrum data is: 13 C NMR (101 MHz, DMSO) δ 193.26 (s), 164.14 (s), 150.36 (s), 149.58 (s), 148.20 (s), 146.37 (s), 142.72 (s), 138.01 (s), 133.55 (s), 131.18 (s), 130.61 (s), 127.34 (d, J = 7.5 Hz), 123.44 (s), 121.69 (s), 120.95 (s), 120.58 (s), 114.68 (s), 108.93 (s), 105.96 (s), 102.58 (s), 75.02 (s), 57.66 (s), 56.02 (d, J = 22.9 Hz), 26.90 (s).

[0027] The hydrogen spectrum data for unsubstituted acetophenone berberine is: 1H NMR (400 MHz, DMSO) δ 10.01 (s, 1H), 8.95 (s, 1H), 8.17 (d, J = 9.2 Hz, 1H), 8.02 - 7.95 (m, 3H), 7.82 (s, 1H), 7.75 - 7.68 (m, 1H), 7.59 (t, J = 7.7 Hz, 2H), 7.11 (s, 1H), 6.18 (s, 2H), 5.90 (s, 2H), 5.01 - 4.89 (m, 2H), 3.95 (s, 3H), 3.27 - 3.20 (m, 3H);

[0028] Carbon spectrum data is: 13 C NMR (101 MHz, DMSO) δ 194.99 (s), 150.36 (s), 149.55 (s), 148.20 (s), 146.28 (s), 142.57 (s), 138.04 (s), 134.49 (d, J = 13.5 Hz), 133.55 (s), 131.18 (s), 129.45 (s), 128.24 (s), 127.28 (s), 123.55 (s), 121.63 (s), 120.93 (s), 120.58 (s), 108.93 (s), 105.96 (s), 102.57 (s), 75.34 (s), 57.64 (s), 55.92 (s), 26.89 (s).

[0029] A preparation method of a new berberine derivative, comprising the following steps: 1) berberine hydrochloride as raw material is pyrolyzed under high temperature and vacuum to obtain berberrubine; 2) berberrubine is subjected to electrophilic addition reaction with 2-bromo-4'-R-phenylacetophenone to obtain the berberine derivative.

[0030] The specific preparation steps are as follows:

[0031] 1) berberine hydrochloride is placed in a 56°C oven and dried for 40 min, while an oil bath is heated to 195°C, then the dried berberine hydrochloride (10 g, 29.75 mmol) is put into a round-bottom flask, the round-bottom flask is connected to an explosion-proof ball and a tap, and then connected to a vacuum pump, and then the round-bottom flask is moved to the oil bath at 195°C, and the reaction is carried out for about 25 min, the yellow powder in the round-bottom flask turns into dark red solid, TLC is used to detect the reaction during the reaction, and after the reaction is detected to be completed, the dark red solid is crushed and loaded into a silica gel chromatography column for purification (eluent DCM:MeOH = 20:1) to obtain the intermediate berberrubine, and the yield is about 32.8%, and the synthesis equation of the intermediate berberrubine is as follows:

[0032]

[0033] The hydrogen spectrum data of the intermediate berberrubine is as follows:1 H NMR (400 MHz, DMSO) δ 9.15 (s, 1H), 8.06 (s, 1H), 7.62 (s, 1H), 7.30 (d, J = 7.9 Hz, 1H), 6.97 (s, 1H), 6.49 (d, J = 7.9 Hz, 1H), 6.10 (s, 1H), 4.52 (t, J = 6.0 Hz, 1H), 3.76 (s, 2H), 3.07 (d, J = 6.0 Hz, 1H), confirmed to be consistent with the structure described above.

[0034] 2) Take the intermediate berberine red obtained by reaction in 1), dissolve in DMF, and place in an 85°C oil bath to react, so that the berberine red is fully activated; 3) Take 2-bromo-4'-R-phenylacetone and add it to the reaction bottle to react, until a large amount of yellow solid suspension appears, and TLC is used to detect the reaction process, and when the reaction is completed, a proper amount of acetone is added to the reaction bottle to precipitate more solids in the reaction solution, and after standing for 3 min, the solid is filtered, and then washed with 30 mL of water, 30 mL of anhydrous ethanol and 20 mL of acetone in sequence, and dried, to obtain the berberine derivative, and the synthesis equation is shown as follows:

[0035]

[0036] The application of a new berberine derivative includes the application of the original drug and the preparation in sterilization and inhibition of growth of bacteria, and can be specifically used for preventing and treating crop diseases.

[0037] Compared with the prior art, the application has the following advantages and beneficial effects:

[0038] The new berberine derivative provided by the embodiment of the application has good bacteriostatic effect and is obviously superior to the bacteriostatic effect of berberine itself, and can be widely applied to the prevention and treatment of crop diseases. BRIEF DESCRIPTION OF DRAWINGS

[0039] In order to more clearly illustrate the technical scheme of the exemplary embodiments of the application, the drawings required in the embodiments will be briefly introduced below, and it should be understood that the following drawings only show some embodiments of the application, and therefore should not be regarded as a limitation on the scope, and other related drawings can also be obtained by those skilled in the art without creative labor on the basis of these drawings.

[0040] Figure 1 The hydrogen spectrum of p-methoxyphenylacetone berberine provided for the embodiment 1 of the application;

[0041] Figure 2 The carbon spectrum of p-methoxyphenylacetone berberine provided for the embodiment 1 of the application;

[0042] Figure 3A hydrogen spectrum of p-nitroacetophenone berberine provided for the embodiment 2 of the present application;

[0043] Figure 4 A carbon spectrum of p-nitroacetophenone berberine provided for the embodiment 2 of the present application;

[0044] Figure 5 A hydrogen spectrum of unsubstituted acetophenone berberine provided for the embodiment 3 of the present application;

[0045] Figure 6 A carbon spectrum of unsubstituted acetophenone berberine provided for the embodiment 3 of the present application;

[0046] Figure 7 A hydrogen spectrum of intermediate berberine red provided for the present application;

[0047] Figure 8 A nuclear magnetic spectrum of p-fluoroacetophenone berberine provided for the embodiment 4 of the present application;

[0048] Figure 9 A mass spectrum of p-fluoroacetophenone berberine provided for the embodiment 4 of the present application;

[0049] Figure 10 A chromatogram of p-fluoroacetophenone berberine provided for the embodiment 4 of the present application;

[0050] Figure 11 A nuclear magnetic spectrum of p-chloroacetophenone berberine provided for the embodiment 5 of the present application;

[0051] Figure 12 A mass spectrum of p-chloroacetophenone berberine provided for the embodiment 5 of the present application;

[0052] Figure 13 A chromatogram of p-chloroacetophenone berberine provided for the embodiment 5 of the present application;

[0053] Figure 14 A nuclear magnetic spectrum of p-hydroxyacetophenone berberine provided for the embodiment 6 of the present application;

[0054] Figure 15 A mass spectrum of p-hydroxyacetophenone berberine provided for the embodiment 6 of the present application;

[0055] Figure 16 A chromatogram of p-hydroxyacetophenone berberine provided for the embodiment 6 of the present application. DETAILED DESCRIPTION

[0056] In order to make the objectives, technical solutions, and advantages of the present application clearer, below, combined with embodiments, the present application is further explained in detail. The illustrative embodiments of the present application and the explanations thereof are only used to explain the present application, and do not limit the present application.

[0057] In the following description, numerous specific details are set forth to provide a thorough understanding of the present application. However, it will be apparent to one skilled in the art that the present application can be practiced without the specific details. In other instances, well-known methods have not been described in detail in order to avoid obscuring the present application.

[0058] Reference throughout this specification to "one embodiment", "an embodiment", "one example", or "an example" means that a particular feature, structure, or characteristic described in connection with the embodiment or example is included in at least one embodiment of the application. Thus, the appearances of the phrases "in one embodiment", "in an embodiment", "in one example", or "in an example" in various places throughout this specification are not necessarily all referring to the same embodiment or example. Furthermore, the particular features, structures, or characteristics can be combined in any suitable

[0059] Embodiment 1

[0060] The present application provides a preparation method of 4-methoxyacetophenone berberine (hereinafter referred to as compound 1) and the method comprises the following steps:

[0061] 1) 500 mg, 1.55 mmol of the intermediate berberine red obtained in the first step of the reaction and 429.19 mg, 3.25 mmol of anhydrous potassium carbonate were weighed into 10 mL of DMF, and after the temperature of the oil bath pot was raised to 85°C, the reaction bottle was moved into the oil bath pot for reaction for 10 min to fully activate the berberine red;

[0062] 2) 2.120 g, 9.3 mmol of 2-bromo-4-methoxyacetophenone was weighed into the reaction bottle and reacted for about 25 min, and a large amount of yellow solid suspension appeared in the reaction bottle. TLC was used to detect the reaction process, and after the completion of the reaction was detected, an appropriate amount of acetone was added to the reaction bottle to precipitate more solids in the reaction solution. After standing for 3 min, the solid was filtered and washed with 30 mL of water, 30 mL of anhydrous ethanol and 20 mL of acetone in sequence, and dried to obtain the target product with a yield of 89.5%;

[0063] The synthesis equation is as follows:

[0064]

[0065] Crop disease activity determination:

[0066] The growth rate method is used to determine the control effect of the above prepared compound 1 on 17 fungal diseases, specifically including: strawberry anthracnose, citrus gray mold, potato dry rot, cucumber leaf spot, watermelon wilt, fusarium oxysporum watermelon special type, fusarium solani, special fusarium, oilseed rape sclerotinia, cotton wilt, dematium, fusarium oxysporum gourd special type, tomato wilt, fusarium oxysporum (albizia pathogenic bacteria), eggplant root rot, colletotrichum gloeosporioides, and cucumber gray mold.

[0067] The determination steps are as follows:

[0068] (1) First, compound 1 is prepared into a 1% suspension, and the prepared methoxyphenylacetophenone berberine is prepared into a mother liquor with a test design concentration reduced by 10 times;

[0069] (2) 1 mL of the mother liquor and 9 mL of PDA medium are respectively added to a culture dish with a diameter of 9 cm, thoroughly mixed, cooled, and standby; the blank control is 1 ml of sterile water, and the lead compound berberine and the positive control are added;

[0070] (3) Under sterile conditions, a punch is used to take a 4mm diameter fungus cake at the edge of the target bacteria colony, with the mycelium facing down, inoculated in the center of the drug-containing plate, with sterile water containing the corresponding mass concentration and the lead compound as the blank control and the positive control, 3 repeated groups for each treatment, 26℃ dark culture for 4d, the cross method is used to measure the colony diameter, the inhibition rate of the compound on each strain is calculated, and the average value is taken for 3 times.

[0071] (4) Chemical control: pesticide jinda (21.2% pyraclostrobin + 21.2% fluazinam suspension agent).

[0072] The inhibition rate calculation formula is: I =

(D0-Dt) / (D0-4)

[0073] Human enzyme activity determination:

[0074] The activity of compound 2 and compound 4 on human enzymes IDO and TDO is determined, and the determination steps are as follows:

[0075] 1) Preparation of the agent:

[0076] The sample concentration of the compound is 10mM, mixed with DMSO at a ratio of 1:1, and shaken uniformly using a vortex shaker to obtain a 5mM compound solution. Take 5mM of the compound solution and mix with DMSO at a ratio of 1:10, and shake uniformly using a vortex shaker to obtain a 0.5mM compound solution.

[0077] 2) Add compound, enzyme, and substrate

[0078] Add 1.2 μL of compound solution of different concentrations in 96-well plates, 1.2 μL of positive compound in positive wells, 1.2 μL of DMSO in control wells, after adding the compound, add 40 μL of IDO, TDO buffer, 10 μL of protein (IDO 40 nM, TDO 80 nM), 10 μL of substrate (400 μM L-Trp).

[0079] Carry out enzyme reaction, incubate at 37°C for 1 h 10 min, after incubation, add 30 μL of 30% trichloroacetic acid per well, incubate at 50°C for 30 min.

[0080] 3) Color development and determination of absorbance

[0081] After incubation, cool at room temperature, add 90 μL of 2% p-diaminobenzaldehyde acid solution, incubate on a shaker for 5 min, determine absorbance at 480 nm.

[0082] 4) Blank absorbance determination

[0083] Blank absorbance determination is the absorbance of the determination system without adding enzyme and substrate, which is used to exclude the influence of the compound itself and other reagents.

[0084] 5) Data processing

[0085] For each well, the inhibition rate is the difference between the absorbance of the compound in the well and the blank absorbance of the compound, divided by the difference between the control absorbance and the control blank absorbance, in percentage form.

[0086]

[0087] Table 1 Inhibition rate % of p-methoxyacetophenone berberine on two strains at different concentrations

[0088]

[0089] The results in Table 1 show that the inhibition rate of compound 1 on Rhizoctonia solani and Sclerotinia sclerotiorum increases with the decrease of concentration gradient, and the inhibition rate at each concentration is higher than that of the lead compound, which is comparable to the fungicide. Therefore, in view of its application, the activity of other strains at the corresponding concentration is determined, and the determination results are shown in Table 2:

[0090] Table 2 Inhibition rate % of p-methoxyacetophenone berberine on various strains

[0091]

[0092]

[0093] Table 2 shows that compound 1 has a certain inhibitory effect on various strains. Except for the inhibitory rate against specific schistosomes which is lower than that against the lead compound, the inhibitory rate against other pathogens is higher than that against the lead compound. Among them, the inhibitory effect is better against three strains: citrus gray mold, tomato wilt, and cucumber gray mold. The inhibitory effect is the best against rapeseed sclerotium, with an inhibition rate of 82.81%.

[0094] Example 2

[0095] This invention provides a p-nitroacetophenone berberine (hereinafter referred to as compound 2) and its preparation method, comprising the following steps:

[0096] (1) Weigh 500 mg and 1.55 mmol of berberine, the intermediate obtained in the first step reaction, and 429.19 mg and 3.25 mmol of anhydrous potassium carbonate and dissolve them in 10 mL of DMF. After the oil bath temperature is raised to 85 °C, the reaction flask is transferred to the oil bath and reacted for 10 min to fully activate berberine.

[0097] (2) Weigh 2.259 g and 9.3 mmol of 2-bromo-4'-nitroacetophenone and add them to the reaction flask. React for about 25 min. A large amount of yellow solid suspension appears in the reaction flask. During the reaction, TLC is used to detect the reaction. After the reaction is completed, add an appropriate amount of acetone to the reaction flask to precipitate more solid in the reaction solution. Let stand for 3 min, filter, and wash the solid with 30 mL of water, 30 mL of anhydrous ethanol and 20 mL of acetone in sequence. Dry to obtain the target product with a yield of 90%.

[0098] The synthesis equation is as follows:

[0099]

[0100] Crop disease activity assay:

[0101] The bacterial strains and experimental methods used were the same as in Example 1. The results are shown in Tables 3 and 4.

[0102] Table 3. Inhibition rate (%) of compound 2 against different concentrations of two bacterial strains

[0103]

[0104] Table 3 shows that the inhibition rate of compound 2 against Rhizoctonia solani and Sclerotium sclerotium of rapeseed increased with decreasing concentration gradient, reaching 96.02% and 82.46% respectively at a 50-fold dilution. This compound exhibits good antibacterial activity at high concentrations. Therefore, to assess its applicability, activity tests were conducted on other bacterial species at corresponding concentrations, and the results are shown in Table 4.

[0105] Table 4. Inhibition rate (%) of compound 2 against various bacterial strains

[0106]

[0107]

[0108] Table 5 Inhibition rate of compound 2 on two human enzymes IDO and TDO

[0109]

[0110] The results in Table 4 show that compound 2 has certain antibacterial effect on each strain, and the antibacterial rate of compound 2 on other strains is higher than that of the lead compound, and the antibacterial effect of compound 2 on strawberry anthracnose, potato dry rot, tomato wilt, gelatinous anthracnose and cucumber gray mold is better. Compound 2 has better inhibition effect on human enzymes IDO and TDO, and the inhibition rate of compound 2 is higher than that of the lead compound, and the inhibition effect of compound 2 is better than that of the positive compound at 100 μM.

[0111] Example 3

[0112] The embodiment of the present application provides a non-substituted phenylacetone berberine (hereinafter referred to as compound 3) and a preparation method thereof, which comprises the following steps:

[0113] (1) 500 mg, 1.55 mmol of the intermediate berberine obtained in the first step reaction and 429.19 mg, 3.25 mmol of anhydrous potassium carbonate are dissolved in 10 mL of DMF, and the oil bath pot temperature is raised to 85°C, then the reaction bottle is moved to the oil bath pot for reaction for 10 min, so that the berberine is fully activated;

[0114] (2) 1.841 g, 9.3 mmol of 2-bromoacetophenone is added to the reaction bottle and reacted for about 25 min, a large amount of yellow solid suspension appears in the reaction bottle, and TLC is used to detect the reaction during the reaction. After detecting that the reaction is completed, an appropriate amount of acetone is added to the reaction bottle to precipitate more solids in the reaction solution, and after standing for 3 min, the solids are filtered and washed with 30 mL of water, 30 mL of anhydrous ethanol and 20 mL of acetone in sequence, and dried to obtain the target product, and the yield is 89%;

[0115] The synthesis equation is as follows:

[0116]

[0117] Crop disease activity determination: the strains used are 15 strains used in Example 1, and the experimental method is the same as that in Example 1, and the determination results are shown in Tables 6 and 7:

[0118] Table 6 Inhibition rate of compound 3 on two strains at different concentrations

[0119]

[0120] The results in Table 6 show that the antibacterial rate of compound 3 on Rhizoctonia solani and Sclerotinia sclerotiorum increases with the decrease of concentration gradient, and the antibacterial effect on Sclerotinia sclerotiorum is better, and when the dilution concentration is 50 times, the antibacterial rate reaches 92.89%. The compound has good antibacterial activity at high concentration. Therefore, the application is balanced, and the activity of other strains at the corresponding concentration is determined, and the determination results are shown in Table 7:

[0121] Table 7: Antibacterial rate of compound 3 on each strain

[0122]

[0123]

[0124] The results in Table 7 show that compound 3 has certain antibacterial effect on each strain, and the antibacterial rate of the compound on special pointed spores, eggplant root rot and watermelon wilt is lower than that of the lead compound, and the antibacterial rate of the compound on other pathogens is higher than that of the lead compound.

[0125] Example 4

[0126] The embodiment of the present application provides a compound of p-fluoroacetophenone berberine (hereinafter referred to as compound 4) and a preparation method thereof, which comprises the following steps:

[0127]

[0128] Berberine red (500 mg, 1.55 mmol, 1 eq) is dissolved in DMF, potassium carbonate (427.8 mg, 3.1 mmol, 2 eq) and p-fluoroacetophenone (1.4 g, 6.4 mmol, 4 eq) are added, and the reaction is carried out at 85°C. After TLC detection, 10 ml of acetone is added to the reaction solution, and after cooling to room temperature, filtration is carried out, and the target compound 4 is obtained after drying with 20 ml of water, 20 ml of ethanol and 20 ml of acetone.

[0129] Crop disease activity determination: the strains used are cotton wilt, verticillium dahliae and tomato early blight, and the experimental method is the same as in Example 1, and the determination results are shown in Table 8:

[0130] Table 8: Antibacterial rate of compound 4 on each strain

[0131]

[0132]

[0133] Table 5: Inhibition rate of compound 4 on two human enzymes IDO and TDO

[0134]

[0135] The results in Table 8 show that the compound 4 has certain fungistasis on each strain, has good inhibitory effect on 3 disease bacteria and 2 human enzymes, and the inhibitory rate is higher than that of the lead compound, and the inhibitory effect on human enzyme TDO is better, and the inhibitory rate is higher than that of the positive compound.

[0136] Example 5

[0137] The present application provides a kind of p-chloroacetophenone berberine (hereinafter referred to as compound 5) and preparation method thereof, comprising the following steps:

[0138]

[0139] Berberine red (500mg, 1.55mmol, 1eq) is dissolved in DMF, potassium carbonate (427.8mg, 3.1mmol, 2eq), p-chloroacetophenone (1.5g, 6.4mmol, 4eq) is added, and the reaction is carried out at 85 DEG C.TLC detects that the reaction is complete, then 10ml of acetone is added to the reaction solution, and after cooling to room temperature, it is filtered, washed with 20ml of water, 20ml of ethanol and 20ml of acetone, and then dried to obtain the target compound 5.

[0140] Crop disease activity determination: the strains used are cotton wilt, verticillium dahliae and tomato early blight, and the experimental method is the same as in Example 1, and the determination results are shown in Table 9:

[0141] Table 9 Fungistasis rate % of compound 5 on each strain

[0142]

[0143]

[0144] The results in Table 9 show that the compound 5 has certain fungistasis on each strain, and the fungistasis rate on 3 disease bacteria is higher than that of the lead compound.

[0145] Example 6

[0146] The present application provides a kind of p-chloroacetophenone berberine (hereinafter referred to as compound 5) and preparation method thereof, comprising the following steps:

[0147]

[0148] Berberine red base (500 mg, 1.55 mmol, 1 eq) was dissolved in MeCN, potassium carbonate (427.8 mg, 3.1 mmol, 2 eq), p-hydroxyacetophenone (1.8 g, 6 mmol, 4 eq) were added, and the reaction was carried out at 70°C. After the reaction was completed by TLC detection, 10 ml of acetone was added to the reaction solution, and after cooling to room temperature, it was filtered, washed with 20 ml of water, 20 ml of ethanol, and 20 ml of acetone, and dried to obtain the target compound 6.

[0149] Crop disease activity determination: The strains used were cotton wilt, verticillium wilt, and tomato early blight, and the experimental method was the same as in Example 1, and the determination results are as follows:

[0150] Table 10 Inhibition rate % of compound 6 on each strain

[0151]

[0152] Table 10 results show that compound 6 has a certain inhibition effect on each strain, and the inhibition rates on two pathogens (cotton wilt and verticillium wilt) are higher than those of the lead compound, and has no inhibition effect on tomato early blight.

[0153] The above specific embodiments further illustrate the purpose, technical solutions, and beneficial effects of the present application. It should be understood that the above description is only a specific embodiment of the present application and is not intended to limit the protection scope of the present application. Any modification, equivalent replacement, improvement, etc. within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. A berberine derivative, characterized in that, The structural formula is as follows: ； R is CH3O-, named p-methoxyacetophenone berberine, with the following structural formula: ； Alternatively, R is -NO2, named p-nitroacetophenone berberine, with the following structural formula: ； Alternatively, R may be -H, and the name of the product is unsubstituted acetophenone berberine, with the following structural formula: ； Alternatively, R may be -F, and its name is p-fluoroacetophenone berberine, with the following structural formula: ； Alternatively, R may be -Cl, named p-chloroacetophenone berberine, with the following structural formula: ； Alternatively, R may be -OH, and its name is p-hydroxyacetophenone berberine, with the following structural formula: 。 2. A method for preparing the berberine derivative as described in claim 1, characterized in that, The process includes the following steps: 1) Berberine hydrochloride is used as a raw material and is subjected to high-temperature vacuum pyrolysis to obtain berberine red alkaloid; 2) Berberine red alkaloid undergoes an electrophilic addition reaction with 2-bromo-4'-R-acetophenone to obtain berberine derivatives.

3. The method for preparing berberine derivatives according to claim 2, characterized in that, The specific preparation steps are as follows: 1) Dry berberine hydrochloride, put it into a reaction flask, draw a vacuum, and then place it in an oil bath at 195°C to react until the yellow powder turns into a dark red solid, thus obtaining the intermediate berberine; 2) Dissolve the intermediate berberine obtained in 1) and anhydrous potassium carbonate in DMF, and place it in an oil bath at 85°C to fully activate the berberine; 3) Add 2-bromo-4'-R-acetophenone to the reaction flask to react until a large amount of yellow solid suspension appears. After the reaction is completed, filter, wash, and dry to obtain the berberine derivative.

4. The application of the berberine derivative as described in claim 1 or the berberine derivative prepared by the method described in claim 2 or 3 in the sterilization and inhibition of bacterial growth of agricultural crops.

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