The invention relates to (E)-Napos; application of-benzylidene nicotinoyl hydrazine compound

Abstract

The invention discloses an application of an (E)-N '-benzylidene nicotinoyl hydrazine compound, and particularly relates to an application of the (E)-N'-benzylidene nicotinoyl hydrazine compound in preparation of drugs for preventing and treating tobacco mosaic viruses. (E)-N '-benzylidene nicotinoyl hydrazine can inhibit infection of TMV to hosts. The (E)-N '-benzylidene nicotinohydrazide pharmaceutical preparation has a good effect of preventing and treating the tobacco mosaic virus disease, and can be used for preparing the pharmaceutical preparation for preventing and treating the tobacco mosaic virus disease.

Description

Technical Field

[0001] The invention belongs to the technical field of biological applications, and further to the technical field of biological pesticides, and specifically relates to the application of an (E)-N'-benzylidene nicotinic acid hydrazide compound. Background Art

[0002] Plant virus diseases are called plant cancers and are the second largest plant diseases after plant fungal diseases. Tobacco mosaic virus (TMV) is the pathogen of tobacco mosaic disease, etc. It has a wide host range and can infect more than 800 species in 65 families, including tobacco, peppers, tomatoes, eggplants, etc. It is difficult to cure after infecting plants, which can easily cause serious economic losses. my country's agricultural production areas are widely distributed and large in area, resulting in frequent mutations of TMV, the emergence of various symptom types, and prevention and control work has become increasingly difficult. Therefore, there is an urgent need to develop new antiviral agents.

[0003] At present, the most reported effects of nicotine derivatives are the treatment of nervous system diseases, in vitro antibacterial activity and insecticidal activity. (E)-N'-benzylidene nicotinic acid hydrazide is a nicotine derivative designed with nicotine as the parent nucleus. So far, there has been no report on the use of (E)-N'-benzylidene nicotinic acid hydrazide compounds as preparations for the prevention and control of tobacco mosaic disease and in the preparation of anti-tobacco mosaic virus drugs. Summary of the invention

[0004] The object of the present invention is to provide an application of a (E)-N'-benzylidene nicotinic acid hydrazide compound.

[0005] The object of the present invention is achieved by using the (E)-N'-benzylidene nicotinic acid hydrazide compound in the preparation of drugs for preventing and treating tobacco mosaic virus.

[0006] The (E)-N'-benzylidene nicotinic acid hydrazide of the present invention is synthesizable.

[0007] The molecular formula of the (E)-N'-benzylidene nicotinic acid hydrazide is C 13 H 11 N3O, the structural formula is: .

[0008] The compound is named: (E)-N'-benzylidene nicotinic acid hydrazide The invention has been proved by experiments that (E)-N'-benzylidene nicotinic acid hydrazide has a high activity of inhibiting TMV infection. The (E)-N'-benzylidene nicotinic acid hydrazide pharmaceutical preparation of the invention has a good effect on preventing and treating tobacco mosaic virus disease, and can be used to prepare a pharmaceutical preparation for preventing and treating tobacco mosaic virus disease.

[0009] When the compound of the present invention is used as a medicine, it can be used directly or in the form of a pharmaceutical composition. The pharmaceutical composition contains 0.1-99%, preferably 0.5-90%, of the compound of the present invention, and the rest is a pharmaceutically acceptable, inert pharmaceutical carrier and / or excipient. The pharmaceutical carrier or excipient is one or more solid, semi-solid and liquid diluents, fillers and pharmaceutical product adjuvants. Various dosage forms are prepared by methods recognized in the pharmaceutical and food fields, such as liquid preparations (aqueous preparations, emulsions, etc.), solid preparations (tablets, granules, etc.), sprays, aerosols, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1 The fluorescence signal intensity in the leaves of Nicotiana benthamiana at 120 h after inoculation of mixtures of (E)-N'-benzylidene nicotinic acid hydrazide compounds and TMV-CP Agrobacterium with different concentrations; Figure 2 is the anti-TMV activity of different concentrations of (E)-N'-benzylidene nicotinic acid hydrazide compounds; Figure 3 (E)-N'-benzylidene nicotinic acid hydrazide compounds induce the expression of plant defense-related genes. DETAILED DESCRIPTION

[0011] The present invention is further described below in conjunction with the embodiments, but the present invention is not limited in any way. Any changes or substitutions made based on the teachings of the present invention belong to the protection scope of the present invention.

[0012] The application of the (E)-N'-benzylidene nicotinic acid hydrazide compound of the present invention is the application of the (E)-N'-benzylidene nicotinic acid hydrazide compound in the preparation of drugs for preventing and treating tobacco mosaic virus.

[0013] The concentration of the (E)-N'-benzylidene nicotinic acid hydrazide compound solution in the preparation of the drug for preventing and treating tobacco mosaic virus is 25-100 μg / mL.

[0014] The molecular formula of the (E)-N'-benzylidene nicotinic acid hydrazide compound is C 13 H 11 N3O, the structural formula is as follows: .

[0015] The (E)-N'-benzylidene nicotinic acid hydrazide compound solution is obtained by dissolving a formulated (E)-N'-benzylidene nicotinic acid hydrazide compound in a solvent and stirring and mixing the mixture to dissolve the mixture.

[0016] The solvent is DMSO solution.

[0017] The present invention is further described below with specific implementation cases: Example 1

[0018] Preparation of 0.5 mg / mL (E)-N'-benzylidene nicotinic acid hydrazide aqueous solution: Weigh 1 mg of (E)-N'-benzylidene nicotinic acid hydrazide compound and dissolve it in a small amount of DMSO solution, stir and mix thoroughly, and after it is fully dissolved, add the remaining amount of distilled water to a final volume of 2 mL to prepare an aqueous solution to obtain a 0.5 mg / mL (E)-N'-benzylidene nicotinic acid hydrazide aqueous solution. Dilute it according to the required concentration when using.

[0019] Example 2

[0020] Preparation of 100 μg / mL (E)-N'-benzylidene nicotinic acid hydrazide aqueous solution: Weigh 100 μg of (E)-N'-benzylidene nicotinic acid hydrazide compound and dissolve it in a small amount of DMSO solution, stir and mix thoroughly, and after it is fully dissolved, add the remaining amount of distilled water to a final volume of 1 mL to prepare an aqueous solution to obtain a 100 μg / mL (E)-N'-benzylidene nicotinic acid hydrazide aqueous solution.

[0021] Example 3

[0022] Preparation of 50 μg / mL (E)-N'-benzylidene nicotinic acid hydrazide aqueous solution: Weigh 50 μg of (E)-N'-benzylidene nicotinic acid hydrazide compound and dissolve it in a small amount of DMSO solution, stir and mix thoroughly, and after it is fully dissolved, add the remaining amount of distilled water to a final volume of 1 mL to prepare an aqueous solution to obtain a 50 μg / mL (E)-N'-benzylidene nicotinic acid hydrazide aqueous solution.

[0023] Example 4

[0024] Preparation of 1000 μg / mL (E)-N'-benzylidene nicotinic acid hydrazide aqueous solution: Weigh 1000 μg of (E)-N'-benzylidene nicotinic acid hydrazide compound and dissolve it in a small amount of DMSO solution, stir and mix thoroughly, and after it is fully dissolved, add the remaining amount of distilled water to a final volume of 1 mL to prepare an aqueous solution to obtain a 1000 μg / mL (E)-N'-benzylidene nicotinic acid hydrazide aqueous solution. Dilute it according to the required concentration when using.

[0025] Example 5

[0026] Preparation of 1 g (E)-N'-benzylidene nicotinic acid hydrazide emulsion: Weigh 0.99 g of (E)-N'-benzylidene nicotinic acid hydrazide compound, add a little DMSO to fully dissolve it; then add 10% sodium dodecylbenzene sulfonate as an emulsifier, mix well to form an emulsion. When using, dilute the emulsion to the required concentration.

[0027] Example 6

[0028] Preparation of 1 g (E)-N'-benzylidene nicotinic acid hydrazide tablets: Weigh 0.4 g of (E)-N'-benzylidene nicotinic acid hydrazide compound, add a little DMSO to fully dissolve it; then add 0.1 g wheat protein (powder), add water to fully stir and mix, then add 40% starch as an excipient, mix well, dry and press into tablets. When using, dissolve the tablets in water and dilute to the required concentration.

[0029] Example 7

[0030] Based on the infectious cloning test, the effect of different concentrations of (E)-N'-benzylidene nicotinic acid hydrazide on TMV was investigated: Use a pipette to pick up the tobacco mosaic virus-infectious cloned Agrobacterium (agro-p35s-30B:GFP) glycerol bacteria stored at -80°C into 2 mL of LB liquid culture medium containing antibiotics (rifampicin and kanamycin), and culture at 28°C and 200 RPM for 24 h. After the first activation, take 100 uL of the infection solution into the LB liquid culture medium, and culture at 28°C and 200 RPM for 24 h. The activation is complete. Centrifuge the activated TMV infection solution at 4000 RPM for 2 min to obtain a bacterial precipitate. Use the infection buffer to adjust the OD value of the bacterial precipitate to about 0.8, and let it stand at 28°C for 8 hours before use.

[0031] 50 μL of (E)-N'-benzylidene nicotinic acid hydrazide compound with a concentration of 25 μg / mL, 50 μg / mL, and 100 μg / mL was mixed with 50 μL of bacterial solution as an injection agent. After mixing, it was allowed to stand at room temperature for 30 minutes before injection. The mixture of 1% DMSO and bacterial solution was used as the control group, and 8% Ningnanmycin aqueous solution diluted 1000 times was used as the positive control. Select Nicotiana benthamiana with similar growth, select leaves from the same layer and the same part of each plant, select an injection point on both sides of the leaf vein, and use a 1mL sterile syringe to quantitatively inject from the back of the leaf (100 μL / injection point). Each group treated 3 tobacco plants, and repeated three times.

[0032] 120 hours after injection, the treated tobacco leaves were placed in the iBright 1500 instrument to take photos and record the fluorescence signals. The fluorescence intensity on the leaves was then analyzed based on the iBright Analysis Software software to calculate the inhibition rate of the compound. Inhibition rate (%) = [(fluorescence intensity of the control group - fluorescence intensity of the treatment group) / fluorescence intensity of the control group] × 100% Test results: Figure 1The effect of different concentrations of (E)-N'-benzylidene nicotinic acid hydrazide compounds on TMV shows that the fluorescence intensity gradually weakened with the increase of concentration when treated with (E)-N'-benzylidene nicotinic acid hydrazide compounds, and was significantly weaker than that of the 1% DMSO treatment group and close to that of the Ningnanmycin group. Figure 2 ), the inhibition rate of the Ningnanmycin treatment group was 59.8%, and the inhibition rates of the 25μg / mL, 50μg / mL, and 100μg / mL (E)-N'-benzylidene nicotinic acid hydrazide compound groups were 47%, 48.8%, and 60.9%, respectively. This shows that the ability of (E)-N'-benzylidene nicotinic acid hydrazide compounds to resist TMV gradually increases with the increase in concentration, and 100μg / mL (E)-N'-benzylidene nicotinic acid hydrazide is most suitable for preparing a pharmaceutical preparation for the prevention and treatment of tobacco mosaic virus disease.

[0033] Example 8

[0034] Effects of (E)-N'-benzylidene nicotinic acid hydrazide on plant-related defense genes: Select 5-6 leaves of K326 tobacco with similar growth and health, apply 200μL of 100μg / mL (E)-N'-benzylidene nicotinic acid hydrazide, and use the group applied with 1% DMSO as the control. Three tobacco plants were treated in each group. Samples were collected after 12 hours and ground with liquid nitrogen in a precooled mortar. After RNA extraction, reverse transcription was performed, and relative quantitative analysis of plant defense-related genes was performed using fluorescent quantitative PCR technology.

[0035] The test results are as follows Figure 3 As shown in the data, after treatment with 100 μg / mL of (E)-N'-benzylidene nicotinic acid hydrazide, the relative expression levels of four plant defense-related genes increased, among which RBOHB, PR5, POR1, and PR1 increased by 7.63, 2.15, 2.52, and 2.11 times, respectively. These results indicate that (E)-N'-benzylidene nicotinic acid hydrazide can up-regulate defense-related genes in plants and enhance plant disease resistance, thereby inhibiting the replication and proliferation of TMV in plants.

Claims

1. An application of a (E)-N'-benzylidene nicotinic acid hydrazide compound, characterized in that: The (E)-N'-benzylidene nicotinic acid hydrazide compound is used in the preparation of drugs for preventing and treating tobacco mosaic virus.

2. The use of the (E)-N'-benzylidene nicotinic acid hydrazide compound according to claim 1, characterized in that: The concentration of the (E)-N'-benzylidene nicotinic acid hydrazide compound solution in the preparation of the drug for preventing and treating tobacco mosaic virus is 25-100 μg / mL.

3. The use of the (E)-N'-benzylidene nicotinic acid hydrazide compound according to claim 1 or 2, characterized in that: The molecular formula of the (E)-N'-benzylidene nicotinic acid hydrazide compound is C 13 H 11 N3O, the structural formula is as follows: 。 4. The use of the (E)-N'-benzylidene nicotinic acid hydrazide compound according to claim 2, characterized in that: The (E)-N'-benzylidene nicotinic acid hydrazide compound solution is obtained by dissolving the (E)-N'-benzylidene nicotinic acid hydrazide compound of a formula ratio in a solvent and stirring and mixing the mixture thoroughly to dissolve the mixture.

5. The use of the (E)-N'-benzylidene nicotinic acid hydrazide compound according to claim 4, characterized in that: The solvent is DMSO solution.

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