Biological activator based on amino acid structure modification and synthesis method and application thereof

Abstract

This invention belongs to the field of agrochemical technology, specifically relating to the synthesis and application of a class of bioactivators based on amino acid structure modification. Using readily available and inexpensive naturally derived glycine, L-alanine, L-valine, and L-leucine as raw materials, this invention synthesizes the corresponding target compound, 2-(diethylamino)ethyl amino acid ester, through a one-step esterification reaction for structural modification. Bioactivity experiments show that this type of compound can significantly promote plant germination and enhance root and stem growth at low concentrations. Germination rate, root length, and stem length are significantly better than the blank control, and compared with existing products, it achieves higher growth promotion efficiency at even lower concentrations. Furthermore, this type of compound has a simple production process, low raw material costs, excellent water solubility, and is easy to scale up for industrial production. It is of great significance for reducing agricultural production costs, improving crop yield and quality, and developing green and environmentally friendly bioactivators.

Description

Technical Field

[0001] This invention relates to the field of agrochemical technology, specifically to bioactivators based on amino acid structure modification, their synthesis methods, and applications. Background Technology

[0002] DA-6, a widely used commercial plant growth regulator, is primarily synthesized via a classic esterification reaction using diethylaminoethanol and hexanoic acid as raw materials. However, limited by its basic structural design, the DA-6 molecule contains only two core functional segments: a tertiary amine group and an ester group. Furthermore, the spatial arrangement and electronic effects of these two segments are fixed, resulting in a limited number of modifiable sites and poor structural flexibility. As modern agriculture transitions towards "precision regulation, green and efficient" practices, existing simple structural regulators are insufficient to meet diverse needs. Introducing new active units is a crucial pathway to expand regulatory functions (such as stress resistance and quality improvement).

[0003] Amino acids, as a naturally occurring class of compounds, have the following advantages: 1. Widely available from natural sources (can be extracted from plants or prepared through fermentation), with low raw material costs; 2. The molecular structure contains amino (-NH2) and carboxyl (-COOH) groups, making it easy to undergo structural modifications such as esterification and amination; 3. Excellent water solubility; aqueous solutions can be prepared without the addition of co-solvents, reducing application costs. 4. It has good biocompatibility and can be degraded into harmless substances by microorganisms in the soil, with no risk of residue.

[0004] Currently, there is limited research on ester derivatives with amino acid-guided structures in the field of plant growth regulation, and there are no reports on plant regulation-related derivatives and their biological activities. Developing such compounds has important theoretical and applied value. Summary of the Invention

[0005] To address the problems of existing plant growth regulators such as "low regulation efficiency, high cost, and residue risk," the present invention aims to synthesize 2-(diethylamino)ethyl amino acid esters through a one-step esterification reaction using naturally derived glycine, L-alanine, L-valine, and L-leucine as raw materials. These compounds contain both amino and diethylaminoethoxy groups for specific effects; the diethylaminoethoxy group is a known active group (similar to the active structure of DA-6), which can synergistically enhance regulation efficiency, ultimately achieving high-efficiency growth promotion at low concentrations.

[0006] Another objective of this invention is to provide a method for synthesizing the above-mentioned compounds, as well as their application in crops such as wheat and soybeans, to provide technical support for the development of green agrochemical products.

[0007] To achieve the above-mentioned technical objectives, the technical solution of the present invention is as follows: The structural formula of the bioactivator based on amino acid structure modification is as follows: ; The structural formula of the bioactivator provided by this invention is as follows: ; A method for synthesizing a bioactivator based on amino acid structure modification, the reaction formula is as follows: ; This is achieved through the following preparation steps: N,N-diethylethanolamine and amino acids were mixed and dissolved in toluene. Then, the catalyst was added to the reaction system and heated to react. Water produced in the reaction system was separated by a water separator. After the reaction was completed, the heating was turned off and the reaction system was cooled to room temperature. The organic phase was filtered to remove insoluble matter, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain the target product 2-(diethylamino)ethyl amino acid ester.

[0008] The catalyst is titanium oxysulfate, and the molar ratio of N,N-diethylethanolamine, amino acid and catalyst is 1:(0.8-1.2):(0.04-0.06).

[0009] The heating reaction is carried out at a temperature of 110-130℃ for 4-8 hours.

[0010] The application of bioactivators based on amino acid structure modification allows them to be used as active or activating ingredients to regulate plant growth activity.

[0011] Preferably, it is used for growth regulation of monocotyledonous or dicotyledonous plants.

[0012] Preferably, when used as a bioactivator, the appropriate concentration for promoting the germination and growth of wheat and soybeans is 5-40 ppm.

[0013] Compared with the prior art, the present invention has the following beneficial effects: 1. Based on the principle of superposition of active substances and combined with the structural characteristics of amino acids and amino esters, the compound of this invention contains the active group diethylaminoethoxy of amino esters, and combined with the structural characteristics of amino acids, its structure is mutually supportive and has high atom utilization.

[0014] 2. 2-(diethylamino)ethyl amino acid esters have good water solubility, with a solubility of ≥30 g / L in water at 25℃.

[0015] 3. It has a good effect on the germination and growth of wheat and soybeans, and significantly improves the efficiency of plant regulation activity.

[0016] 4. The preparation method of this invention is simple, easy to scale up, uses inexpensive and readily available raw materials, and has low production and usage costs. Compared with existing plant growth regulators, it requires less dosage, can significantly improve seed activity, promote rooting and germination, and enhance seedling resistance, providing a new option for the development of plant growth regulators and has excellent application prospects. Detailed Implementation

[0017] The technical solution of the present invention will be clearly and completely described below with reference to specific embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Example 1

[0018] This embodiment provides a method for synthesizing a bioactivator based on amino acid structure modification, which is achieved through the following preparation steps: Glycine (0.4 mol), N,N-diethylethanolamine (0.4 mol), toluene (100 mL), and the catalyst titanium oxysulfate (0.02 mol, 0.05 eq) are added to a 250 mL two-necked flask, and then heated to 120 °C for 6 h. Water produced in the reaction system is separated using a water separator. After the reaction is complete, the heating is turned off, and the system is allowed to cool to room temperature. The organic phase is filtered to remove insoluble matter, and the solvent is removed by vacuum distillation. The crude product is purified by silica gel column chromatography (n-hexane / ethyl acetate = 1:1) to obtain the target product 2-(diethylamino)ethylglycine ester, with a yield of 94%. Example 2

[0019] This embodiment provides a method for synthesizing a bioactivator based on amino acid structure modification, which is achieved through the following preparation steps: L-valine (0.32 mol), N,N-diethylethanolamine (0.4 mol), toluene (100 mL), and the catalyst titanium oxysulfate (0.016 mol, 0.04 eq) are added to a 250 mL two-necked flask, and then heated to 120 °C for 6 h. Water produced in the reaction system is separated using a water separator. After the reaction is complete, the heating is turned off, and the system is allowed to cool to room temperature. The organic phase is filtered to remove insoluble matter, and the solvent is removed by vacuum distillation. The crude product is purified by silica gel column chromatography (n-hexane / ethyl acetate = 1:1) to obtain the target product 2-(diethylamino)ethyl-L-valine ester, with a yield of 96%. Example 3

[0020] This embodiment provides a method for synthesizing a bioactivator based on amino acid structure modification, which is achieved through the following preparation steps: L-alanine (0.48 mol), N,N-diethylethanolamine (0.4 mol), toluene (100 mL), and the catalyst titanium oxysulfate (0.024 mol, 0.06 eq) are added to a 250 mL two-necked flask, and then heated to 120 °C for 6 h. Water produced in the reaction system is separated using a water separator. After the reaction is complete, the heating is turned off, and the system is allowed to cool to room temperature. The organic phase is filtered to remove insoluble matter, and the solvent is removed by vacuum distillation. The crude product is purified by silica gel column chromatography (n-hexane / ethyl acetate = 1:1) to obtain the target product 2-(diethylamino)ethyl-L-alanine ester, with a yield of 92.5%. Example 4

[0021] This embodiment provides a method for synthesizing a bioactivator based on amino acid structure modification, which is achieved through the following preparation steps: L-leucine (0.4 mol), N,N-diethylethanolamine (0.4 mol), toluene (100 mL), and the catalyst titanium oxysulfate (0.02 mol, 0.05 eq) are added to a 250 mL two-necked flask, and then heated to 120 °C for 6 h. Water produced in the reaction system is separated using a water separator. After the reaction is complete, the heating is turned off, and the system is allowed to cool to room temperature. The organic phase is filtered to remove insoluble matter, and the solvent is removed by vacuum distillation. The crude product is purified by silica gel column chromatography (n-hexane / ethyl acetate = 1:1) to obtain the target product 2-(diethylamino)ethyl-L-leucine ester, with a yield of 93%.

[0022] 2-(Diethylamino)ethylglycine ester: 1 H NMR (400 MHz, D2O) 4.45(t, J= 4.2 Hz, 2H), d 3.84(s, 2H), 3.38(t, J= 4.2 Hz, 2H), 3.18-3.05(m, 4H), 1.14 (t, J= 4.2 Hz, 6H). 13 C NMR (400 MHz, D2O) 166.7, 60.4, 47.9, 39.6, 8.2. HRMS (ESI): [M+H] + calcd for 175.1368, found 175.1372. 2-(Diethylamino)ethyl-L-alanine ester: 1 H NMR (400 MHz, D2O) d4.46-4.35(m, 2H), 4.35-4.25(m, 1H), 3.4(t, J= 4.2 Hz, 2H), 3.15-3.05(m, 4H), 1.4 (d, J= 4.2 Hz,2H), 1.12 (t, J= 5.2 Hz, 6H). 13 C NMR (400 MHz, D2O) 170.2, 60.7, 55.4, 53.4,49.8, 48.5, 47.3, 15.0, 8.7. HRMS (ESI): [M+H] + calcd for 189.1525, found189.1526. 2-(Diethylamino)ethyl-L-valine ester: 1 H NMR (400 MHz, D2O) d 0.86 (dd, J= 5.2 Hz, 6H). 13 C NMR (400 MHz, D2O)169.2, 60.7, 55.4, 53.4, 50.1, 48.5, 47.6, 15.2, 8.5. HRMS (ESI): [M+H] + calcdfor 217.1838, found 217.1836. 2-(Diethylamino)ethyl-L-leucine ester: 1 H NMR (400 MHz, D2O) d 4.53-4.32(m, 2H),4.08-3.98(m, 1H), 3.44-3.32(m, 2H), 3.22-3.04(m, 4H), 1.76-1.64(m, 1H), 1.63-1.48(m, 2H), 1.14 (t, J= 4.2 Hz, 6H), 0.71 (dd, J= 5.2 Hz, 6H). 13 C NMR (400MHz, D2O) 169.9, 66.4, 60.4, 51.2, 49.7, 47.9, 36.5, 23.8, 21.6, 8.0. HRMS(ESI): [M+H] +calcd for 231.1994, found 231.1995.

[0023] Effect Examples Bioactivity Test

[0024] Preparation of amino acid ester activators: 0.5 g of amino acid ester and DA-6 were weighed and diluted with tap water to 500 mL to obtain a 1 g / L activator stock solution. This stock solution was then diluted to 1 ppm, 5 ppm, 10 ppm, 20 ppm, 40 ppm, 60 ppm, and 80 ppm activator solutions, respectively. Tap water treatment served as the control (CK).

[0025] Seedling cultivation: Wheat and soybean seeds of uniform size and plumpness were selected and placed in 25 mL beakers, with 30 seeds in each beaker. The prepared activator was poured into the beakers containing the seeds, and the seeds were soaked for 6 hours. After soaking, the seeds were evenly arranged on a moist paper bed for germination until the taproot showed white and the lateral roots just began to emerge. During this process, 1.0 g of agar was accurately weighed using an analytical balance and placed in a beaker containing 2000 mL of water. The water was heated in a microwave oven until completely dissolved, cooled, and then poured evenly into four 500 mL beakers until solidified, preparing the agar medium. The germinated seeds were then planted on the solidified agar and cultured in a 27 ℃ incubator. When the taproot of the control group (CK group) seeds touched the bottom of the beaker, all seedlings were removed and measured.

[0026] Measurement indicators and methods Seed germination rate: Number of germinated seeds / Total number of seeds Seedling sampling method: All seedlings were randomly sampled, with 10 seedlings randomly selected from each treatment as test samples, and the samples were repeated 3 times to detect the morphological and physiological indicators of the seedlings.

[0027] Plant height: The standard measurement is the length from the highest point of the leaf to the hypocotyl.

[0028] Root length: The measurement method is to select the three longest roots from a seedling and take their average value.

[0029] Dry weight of above-ground and underground parts: The measurement method is to wash the seedlings clean, absorb the surface moisture with filter paper, and divide them into above-ground and underground parts. They are blanched at 105 ℃ for 20 minutes, and then dried at 80 ℃ to constant weight. The dry weight is then measured.

[0030] The results of the bioactivity tests of amino acid esters are shown in Tables 1-2. ; ; ;

[0031] Bioactivity experiments were conducted on wheat and soybean, respectively, and the results showed that: 1. Using the "seed soaking method" to test wheat germination and growth, it was found that this compound had a better effect on promoting wheat growth at 5-40 ppm, and its effect on promoting germination and growth was far superior to the blank control and DA-6 group; 2. Soybean germination and growth experiments were conducted using the "soaking method". It was found that the compound had a better effect on promoting soybean growth at concentrations of 5-40 ppm, and its effect on promoting germination and growth was far superior to that of the blank control and the DA-6 group.

[0032] This invention synthesizes 2-(diethylamino)ethyl amino acid esters from glycine, L-alanine, L-valine, and L-leucine, respectively. The process is simple and low-cost, and it has the advantages of "high efficiency regulation, good water solubility, and no residue". It has shown better growth regulation effects than existing commercial growth regulators (DA-6) in crops such as wheat and soybeans, providing a new direction for the development of green agrochemical products and has significant agricultural application value and market potential.

[0033] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A bioactivator based on amino acid structure modification, characterized in that, Its structural formula is:

2. The bioactivator based on amino acid structure modification according to claim 1, characterized in that... The structural formula is:

3. The method for synthesizing the bioactivator based on amino acid structure modification as described in claim 1, characterized in that... The preparation process is as follows: N,N-diethylethanolamine and amino acids are mixed and dissolved in toluene. Then, the catalyst is added to the reaction system and heated to react. Water produced in the reaction system is separated by a water separator. After the reaction is completed, the heating is turned off and the reaction system is cooled to room temperature. The organic phase is filtered to remove insoluble matter, the solvent is removed by vacuum distillation, and the crude product is purified by silica gel column chromatography to obtain the target product 2-(diethylamino)ethyl amino acid ester.

4. The method for synthesizing bioactivators based on amino acid structure modification according to claim 3, characterized in that: The catalyst is titanium oxysulfate, and the molar ratio of N,N-diethylethanolamine, amino acid and catalyst is 1:(0.8-1.2):(0.04-0.06).

5. The method for synthesizing bioactivators based on amino acid structure modification according to claim 3, characterized in that: The heating reaction is carried out at a temperature of 110-130℃ for 4-8 hours.

6. The application of the bioactivator based on amino acid structure modification as described in claim 1, characterized in that: It can be used as an active ingredient or activator to regulate plant growth activity.

7. The application of the bioactivator based on amino acid structure modification according to claim 6, characterized in that, It can be used to regulate the growth of monocotyledonous or dicotyledonous plants.

8. The application of the bioactivator based on amino acid structure modification according to claim 6, characterized in that, When used as a bioactivator, the appropriate concentration for promoting the germination and growth of wheat and soybeans is 5-40 ppm.

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