N-aryl amino-acid ester derivative and application thereof in resisting tobacco mosaic virus

Abstract

The invention discloses an N-aryl amino-acid ester derivative and a preparation method and application thereof, the structural formula of the N-aryl amino-acid ester derivative is shown in the specification. Practice proves that the compound has good anti-plant virus activity, the preparation method is simple, raw materials are cheap and easy to obtain, and the N-aryl amino-acid ester derivative is suitable for industrial amplification; along with the increase of the drug resistance of the existing tobacco mosaic virus resisting drug, the compound can be used as an efficient substitute product for preparing an active component of a novel tobacco mosaic virus resisting inhibitor, is suitable for preparing the tobacco mosaic virus resisting drug and the active component thereof, and has obvious application prospects in the related fields of biological drug technology and the like.

Description

Technical Field

[0001] This invention belongs to the field of fine chemical preparation technology, and more specifically, relates to a... N - Aromatic amino acid ester derivatives and their application against tobacco mosaic virus. Background Technology

[0002] Plant viral diseases are a major category of crop diseases. In agricultural production, viral diseases are the second largest category of plant diseases after fungal diseases, and they are devastating diseases affecting crops such as rice, tobacco, and vegetables, often referred to as "plant cancer." Tobacco mosaic virus (TMV) is one of the most prevalent and damaging viruses globally, occurring in all provinces of my country. It can be transmitted through sap, is stable in vitro, and severely affects crop quality after infection, posing a huge threat to agricultural production. Globally, TMV causes hundreds of millions of dollars in losses annually. Because plants lack an immune system like animals, the control of plant viral diseases is relatively difficult and has always been a hot and challenging issue in plant disease and virology research. Currently, compared to the control of pests, weeds, fungi, and bacteria, the effective control and treatment of plant viral diseases is relatively lagging. Besides screening for disease-resistant genes and breeding disease-resistant varieties, antiviral agents remain the most direct and effective means of controlling plant viral diseases. To date, only a few commercially available agents are used to control plant viral diseases. Figure 1 Furthermore, the environmental pollution, increased pathogen resistance, and agricultural product quality and safety problems caused by the long-term irrational use of commercial plant virus inhibitors are becoming increasingly serious. Therefore, given the increasingly severe occurrence of plant viral diseases and the continued scarcity of antiviral agents, in-depth research on antiviral drugs for plants is urgently needed.

[0003]

[0004] Representative commercial plant virus inhibitors Therefore, the industry urgently needs to find a drug molecule that has a short process, high yield, and good activity against tobacco mosaic virus. Summary of the Invention

[0005] One object of the present invention is to provide N - An aryl amino acid ester derivative, the structure of which is shown in structural formula I:

[0006] in: R 1 It is one of -Bn, -Ph, or -H; R 2 For -Me, -Et, - iOne of Pr-, -Bu, -Bn, -CH2CH2OMe; R 3 For -Me, -Et, - i One of Pr-, -Bu, -Bn, or -CH2CH=CH2; Its preparation method includes the following steps:

[0007] Preferably, in the preparation method, an amino acid ester is used as the starting material, which is reacted with bromoacetaldehyde diethanol and a 1,3-dicarbonyl compound to obtain carbazole ester intermediate 2. The reaction solvent is one of anhydrous methanol, anhydrous ethanol, toluene, and acetonitrile, and the reaction catalyst is at least one of aluminum chloride, elemental iodine, zinc chloride, and zirconium chloride. Preferably, in the preparation method, the material ratio of amino acid ester to bromoacetaldehyde diethanol and 1,3-dicarbonyl compound is 1:1:1, the reaction temperature is 50-80℃, and the reaction time is 2-8h.

[0008] The present invention also provides the above. N The application of aryl amino acid ester derivatives in anti-tobacco mosaic virus drugs is characterized by the following: N - An aryl amino acid ester derivative dissolved in a diluent yields an antiviral drug for plants; the antiviral preparation for tobacco mosaic virus contains... N The aryl amino acid ester derivative comprises 0.001–99.99% by weight, wherein the diluent is one or more of propanol, isopropanol, glycerol, toluene, xylene, chlorobenzene, 1,2-dichloroethane, DMSO, ethyl acetate, acetone, butanone, and paraffin; or one or more of talc, dolomite, quartz, kaolin, bentonite, clay, diatomaceous earth, montmorillonite, activated clay, calcium carbonate, oxide film, and magnesia silicate; or one or more of alkyl sulfonates, alkyl sulfonates, alkyl aryl sulfonates, sorbitol polyoxyethylene ester, arylalkyl polyethylene glycol ether, fluoroalkyl sulfonates, alkyl sulfates, and lignin sulfonates; or one or more of polyvinyl alcohol, carboxymethyl cellulose, and gum arabic. Preferably, the N The application of aryl amino acid ester derivatives in the preparation of drugs against tobacco mosaic virus, characterized in that: the formulation contains... N -The weight percentage of aryl amino acid ester derivatives is 5-40%.

[0009] More preferably, the N - The application of aryl amino acid ester derivatives in the preparation of drugs against tobacco mosaic virus is as follows: water emulsion, suspension, wettable powder or water-dispersible granules.

[0010] This invention provides a novel type of N These compounds are aryl amino acid ester derivatives and exhibit excellent anti-tobacco mosaic virus activity. Their preparation methods are simple, the raw materials are inexpensive and readily available, making them suitable for industrial scale-up. With the increasing resistance to existing anti-tobacco mosaic virus drugs, these compounds can serve as highly efficient alternatives for preparing novel plant virus inhibitors. They are suitable for preparing anti-plant virus drugs and their active ingredients, showing significant application prospects in biopharmaceutical technology and related fields. Attached Figure Description

[0011] Figure 1 The 1H NMR spectrum of the product obtained from 4a is shown.

[0012] Figure 2 The 1H NMR spectrum of the product obtained from 4a is shown.

[0013] Figure 3 The high-resolution mass spectra of the product obtained for 4a.

[0014] Figure 4 The 1H NMR spectrum of the product obtained from 4b is shown.

[0015] Figure 5 The 1H NMR spectrum of the product obtained from 4b is shown.

[0016] Figure 6 The high-resolution mass spectra of the product obtained for 4b.

[0017] Figure 7 The 1H NMR spectrum of the product obtained from 4C is shown.

[0018] Figure 8 The 1H NMR spectrum of the product obtained from 4C is shown.

[0019] Figure 9 High-resolution mass spectrometry of the product obtained from 4C. Detailed Implementation

[0020] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention. Furthermore, the technical features involved in the various embodiments of this invention described below can be combined with each other as long as they do not conflict with each other.

[0021] The above scheme will be described in detail below under different reaction conditions.

[0022] Example 1: N Synthesis of aryl amino acid ester derivative 4a 1 mol (0.3 mmol) of amino acid ester L-Val-OMe and 0.45 mmol of diethyl bromoacetaldehyde were dissolved in dichloroethane, an organic solvent. Then, 0.3 mmol of ethyl acetoacetate and 0.15 mol of elemental iodine were added. After purging the reaction system with nitrogen three times, the reaction solution was reacted at 50°C for 5 hours in a reactor equipped with a magnetic stirrer. After the raw materials were consumed as monitored by liquid chromatography, the solvent was removed by vacuum distillation, and the product was separated by column chromatography to obtain a yellow oily product 4a (36.4 mg, 64% yield, 97% ee). (eluting solution: petroleum ether / ethyl acetate = 40 / 1 (v / v)); 1 H NMR (600 MHz, CDCl) 3, TMS, 25 °C δ = 6.77 (s, 1H), 6.26 (s, 1H), 4.41 – 4.31 (m, 4H), 3.88 (d, J = 5.8 Hz, 1H), 3.72 (s, 3H), 2.35 (s, 3H), 2.24 (s, 3H), 2.21 – 2.16(m, 1H), 1.41 – 1.34 (m, 6H), 1.05 (d, J = 6.8 Hz, 3H), 1.01 (d, J = 6.8 Hz, 3H)ppm. 13 C{1H} NMR (151 MHz, CDCl3) δ 173.4, 170.1, 169.1, 148.4, 139.5, 137.2,125.0, 113.4, 110.7, 62.1, 60.9, 60.8, 52.0, 31.4, 20.6, 19.7, 19.1, 18.7,14.3, 14.2. HRMS (ESI): calcd for C 20 H 30 NO6 + [M+H] + : 380.2068; found: 380.2071. Example 2 1 mol (0.3 mmol) of amino acid ester L-Val-OEt and 0.45 mmol of diethyl bromoacetaldehyde were dissolved in the organic solvent dichloroethane. Then, 0.3 mmol of ethyl acetoacetate and 0.15 mol of elemental iodine were added. After purging the reaction system with nitrogen three times, the reaction solution was reacted at 50°C for 2 hours in a reactor equipped with a magnetic stirrer. After the raw materials were consumed as monitored by liquid chromatography, the solvent was removed by vacuum distillation, and the product was separated by column chromatography to obtain a yellow oily product 4b (45.7 mg, 69% yield). (45.7 mg, eluting solution: petroleum ether / ethyl acetate = 40 / 1 (v / v)). 1 HNMR (600 MHz, CDCl 3, TMS, 25 °C δ = 7.29 – 7.25 (m, 3H), 7.24 – 7.21 (m, 1H), 7.18 – 7.15 (m, 2H), 6.65 (d, J = 7.5 Hz, 1H), 6.25 (s, 1H), 4.35 – 4.30 (m,5H), 4.19 – 4.15 (m, 1H), 4.12 – 4.07 (m, 1H), 3.17 (dd, J = 13.7, 6.0 Hz, 1H), 3.09 (dd, J = 13.7, 6.6 Hz, 1H), 2.33 (s, 3H), 2.22 (s, 3H), 1.36 (t, J = 7.1 Hz, 3H), 1.32 (t, J = 7.1 Hz, 3H), 1.19 (t, J = 7.1 Hz, 3H) ppm. 13 C{1H} NMR (151 MHz, CDCl3) δ 172.4, 170.1, 168.7, 147.1, 139.3, 137.0, 136.2, 129.4, 128.5,127.0, 125.1, 114.1, 110.8, 61.2, 60.9, 60.8, 57.5, 38.6, 20.5, 19.5, 14.3,14.2, 14.1. HRMS (ESI): calcd for C 25 H 32 NO6 + [M+H]+ : 442.2224; found: 442.2220. Example 3 Take amino acid ester L-Val- i 1 mol (0.3 mmol) of Pr and 0.45 mmol of diethyl bromoacetaldehyde were dissolved in the organic solvent dichloroethane. Then, 0.3 mmol of ethyl acetoacetate and 0.15 mol of elemental iodine were added. After purging the reaction system with nitrogen three times, the reaction solution was reacted at 50°C for 2 hours in a reactor equipped with a magnetic stirrer. After the raw material was consumed as monitored by liquid chromatography, the solvent was removed by vacuum distillation, and the product was separated by column chromatography to obtain a yellow oily product 4c (50.5 mg, 74% yield). (50.5 mg, eluting solution: petroleum ether / ethyl acetate = 45 / 1 (v / v)). 1 HNMR (600 MHz, CDCl 3, TMS, 25 °C δ = 7.20 – 7.18 (m, 2H), 7.16 – 7.13 (m, 1H), 7.12 – 7.09 (m, 2H), 6.53 (s, 1H), 6.19 (s, 1H), 4.92 – 4.88 (m, 1H), 4.28 –4.23 (m, 4H), 4.21 (s, 1H), 3.07 (dd, J = 13.6, 6.4 Hz, 1H), 3.01 (dd, J = 13.6,6.5 Hz, 1H), 2.26 (s, 3H), 2.14 (s, 3H), 1.28 (t, J = 7.1 Hz, 3H), 1.25 (t, J =7.1 Hz, 3H), 1.12 (d, J = 6.3 Hz, 3H), 1.02 (d, J = 6.3 Hz, 3H) ppm. 13C{1H} NMR(151 MHz, CDCl3) δ 171.9, 168.7, 167.0,147.2, 139.2, 137.0, 136.3, 129.4,128.4, 126.9, 125.1, 114.3, 110.9, 68.9, 60.8, 60.7, 57.7, 38.6, 21.7, 21.5,20.5, 19.3, 14.2, 14.1. HRMS (ESI): calcd for C 26 H 34 NO6 + [M+H] + : 456.2381;found: 456.2397. Example 4: Anti-tobacco mosaic virus activity of compounds 4a, 4b and 4c After accurately weighing compounds 4a, 4b, and 4c, dissolve them in DMSO solution to prepare a 5 × 10 μg / mL stock solution. Then, dilute with an aqueous solution containing 0.1% Tween 80 to 100 μg / mL and 500 μg / mL respectively (i.e., 100 μg / mL each of compounds 4a, 4b, or 4c; 500 μg / mL each of compounds 4a, 4b, or 4c). Ribavirin is used by directly diluting the formulation with water. 1. In vivo passivation Select uniformly growing *Symplocos spp.* at the 4-6 leaf stage. Mix the 100 μg / mL and 500 μg / mL concentrations of compounds 4a, 4b, and 4c prepared above with TMV virus particle solution and incubate for 30 min, then inoculate by friction. The virus concentration is 20 μg / mL. The half-leaf method is used for determination. The right side of the same leaf is inoculated with the mixture of compounds and virus, while the left side is inoculated with the same concentration of virus and 0.1% Tween 80 aqueous solution as a control. After inoculation, rinse with running water. Each treatment is repeated three times. After 4-5 days, the number of leaf lesions is counted, and the inhibition rate is calculated: Inhibition rate (%) = [(Number of control lesions - Number of treated lesions) / Number of control lesions] × 100%. The treatment methods for ribavirin, ningnanmycin, and compounds 4a, 4b, and 4c at 100 μg / mL and 4a, 4b, and 4c at 500 μg / mL are the same.

[0023] 2. In vivo protection Selected uniformly grown *Symplocos spp.* at the 4-6 leaf stage. Using the half-leaf method, the right side of the same leaf was sprayed with the same agent (i.e., 100 μg / mL of compound 4a, 100 μg / mL of compound 4b, and 100 μg / mL of compound 4c; and 500 μg / mL of compound 4a, 500 μg / mL of compound 4b, and 500 μg / mL of compound 4c), while the right side was sprayed with a 0.1% Tween 80 aqueous solution as a blank control. 24 hours later, 500-mesh emery was sprinkled on the leaf surface, and the entire leaf was inoculated with TMV virus using a brush dipped in the virus solution. The virus concentration was 10 μg / mL. The leaves were rinsed with running water after inoculation. Each treatment was repeated three times. The number of leaf lesions was counted after 4-5 days, and the inhibition rate was calculated. 3. In vivo therapeutic effects Select uniformly growing *Nicotiana sambac* plants at the 4-6 leaf stage and inoculate the entire leaf with TMV virus using a brush at a concentration of 10 μg / mL. Rinse with running water 2 hours after inoculation. After the leaves have dried, use the half-leaf method for determination. Spray the right side of the same leaf with the appropriate agent (i.e., spray 100 μg / mL of compound 4a, 100 μg / mL of compound 4b, 100 μg / mL of compound 4c, 500 μg / mL of compound 4a, 500 μg / mL of compound 4b, and 500 μg / mL of compound 4c), and spray the right side with a 0.1% Tween 80 aqueous solution as a control. Each treatment is repeated three times. After 4-5 days, count the number of leaf lesions and calculate the inhibition rate.

[0024] Table 1 N Results of anti-tobacco mosaic virus activity of aryl amino acid ester derivative 4a

[0025] Note: The testing methods for products 4b and 4c are the same as those for product 4a in Example 1.

[0026] As shown in Table 1, compounds 4a, 4b, and 4c all exhibited good inhibitory activity against tobacco mosaic virus (TMV) under all three modes of action. Compounds 4a, 4b, and 4c showed the strongest inactivation activity against TMV, followed by therapeutic activity. Furthermore, at the same concentration, compounds 4a, 4b, and 4c demonstrated superior in vivo inactivation and therapeutic activity against TMV compared to the commercially available agent ribavirin, and slightly higher than the commercially available agent ningnanmycin. In summary, N -Aryl amino acid ester derivatives 4a, 4b and 4c are novel viral inhibitors with great potential for development and application.

[0027] Those skilled in the art will readily understand that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A kind N -Aryl amino acid ester derivative, characterized in that: N - the structure of the aryl amino acid ester derivative is shown in structural formula I: Wherein: R 1 is one of -Bn, -Ph, -H; R 2 is -Me, -Et, - i one of -Pr, -Bu, -Bn, -CH2CH2OMe; R 3 is -Me, -Et, - i one of -Pr, -Bu, -Bn, -CH2CH=CH2.

2. The method of claim 1, wherein the method further comprises: N A method for preparing an aryl amino acid ester derivative, characterized by: In the preparation method, an amino acid ester is used as a starting material, reacted with bromoacetaldehyde diethyl acetal and a 1,3-dicarbonyl compound in a solvent and a catalyst to obtain N - arylamino acid ester derivatives, the reaction formula being as follows: 。 3. The method of claim 2, wherein the method further comprises: N A method for preparing an aryl amino acid ester derivative, characterized by: In the structural formula, R 1 R is any one of -Bn, -Ph, and -H. 2 For -Me, -Et, - i Any one of Pr-, -Bu, -Bn, -CH2CH2OMe; R3 is -Me, -Et, - i Any one of Pr-, -Bu, -Bn, or -CH2CH=CH2.

4. The method of claim 2, wherein the method further comprises: N A method for preparing an aryl amino acid ester derivative, characterized by: The reaction solvent is one of anhydrous methanol, anhydrous ethanol, toluene, acetonitrile, and the reaction catalyst is at least one of aluminum chloride, elemental iodine, zinc chloride, and zirconium chloride.

5. The method of claim 3 N - aryl amino acid ester derivatives characterized by: In the preparation method, the material ratio of amino acid ester, bromoacetaldehyde diethyl acetal and 1,3-dicarbonyl compound is 1:1:1, the reaction temperature is 50-80 DEG C, and the reaction time is 2-8h.

6. An antitobamovirus medicament, characterized in that, The medicament is as defined in claim 1 N - aryl amino acid ester derivatives.

7. The antitobamovirus drug according to claim 6, characterized in that, The medicaments include the compound of claim 1 N - aryl amino acid ester derivatives; The tobacco leaf is dipped into the solution of the aryl amino acid ester derivative, and then dried to obtain the tobacco leaf containing the aryl amino acid ester derivative. N The aryl amino acid ester derivative is dissolved in a diluent to obtain an anti-tobacco mosaic virus drug, and the anti-tobacco mosaic virus preparation contains N The aryl amino acid ester derivative accounts for 0.001-99.99% by weight, and the diluent includes any one of a solvent, a filler, a surfactant and a thickening agent. The tobacco mosaic virus includes TMV virus.

8. The anti-tobacco mosaic virus drug according to claim 7, characterized in that, The solvent includes one or more of propanol, isopropanol, glycerol, toluene, xylene, chlorobenzene, 1,2-dichloroethane, DMSO, ethyl acetate, acetone, butanone, and paraffin wax; The filler includes one or more of talc, dolomite, quartz, kaolin, bentonite, clay, diatomite, montmorillonite, activated white clay, calcium carbonate, oxide film, and silicon-magnesium earth; The surface active agent includes one or more of alkyl sulfonate, alkyl sulfonate, alkyl aryl sulfonate, sorbitol polyoxyethylene ester, aralkyl polyethylene glycol ether, fluorinated alkyl sulfonate, alkyl sulfate, and lignin sulfonate; The thickening agent includes one or more of polyvinyl alcohol, carboxymethyl cellulose, and gum arabic.

9. The antitobamovirus drug according to claim 7, characterized in that: The preparation contains N The aryl amino acid ester derivative accounts for 5-40% by weight; the anti-plant virus drug is an emulsion in water, a suspension, a wettable powder or a water dispersible granule.

10. The method of claim 1 N - Use of an aryl amino acid ester derivative or a tobacco mosaic virus drug according to any one of claims 6 to 9 for the preparation of an agent against plant viruses.

Images