Biological activators of trisubstituted l-proline structures and methods of synthesis and use thereof
The synthesis of triethanolamine tri-L-proline ester via a one-step esterification reaction solves the problems of low regulatory efficiency and residue risk of existing bioactivators, realizing a high-efficiency, low-cost green agrochemical product that promotes the growth of wheat and soybeans.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- HIGH & NEW TECH RES CENT OF HENAN ACAD OF SCI
- Filing Date
- 2026-04-15
- Publication Date
- 2026-07-10
AI Technical Summary
Existing bioactivators have low regulation efficiency, high cost, and pose a risk of soil residue, while there is a lack of green and environmentally friendly multi-active group regulators.
Triethanolamine tri-L-proline ester was synthesized from L-proline via a one-step esterification reaction. This method combines three active sites to enhance molecular permeability and target binding specificity, thus simplifying the synthesis process.
It significantly improves regulation efficiency, reduces costs, shortens reaction time, and achieves efficient promotion of wheat and soybean growth, with no soil residue and high environmental safety.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of agrochemical technology, specifically to bioactivators with trisubstituted L-proline structures, their synthesis methods, and applications. Background Technology
[0002] Bioactivators are key agrochemical products that regulate crop growth and development and improve agricultural productivity. They can enhance crop resistance to stress and increase yield and quality by regulating processes such as seed germination, root development, and organ differentiation. Currently, commercially available regulators, such as DA-6 and sodium nitrophenolate, are widely used, but they have the following shortcomings: some products have a simple structure (containing only a single active group), resulting in limited regulatory efficiency; some synthetic raw materials rely on imports, leading to high costs; and a few products have long residual periods in the soil, posing environmental risks.
[0003] L-proline is a natural cyclic imine widely found in plants and microorganisms, and it has unique advantages as a raw material for activators: 1. It has a wide range of natural sources and can be prepared through microbial fermentation or plant extraction. The purity of industrial-grade products reaches over 98%, and the raw material cost is only 1 / 3 of that of artificially synthesized active monomers. 2. It has a special molecular structure, containing amino (-NH) and carboxyl (-COOH) groups, and has the stability of a cyclic structure. The branched alkyl groups can enhance the permeability of the compound in plant cell membranes, and the carboxyl groups can easily introduce active groups through esterification reactions to achieve "activity superposition". 3. It has excellent biocompatibility and can be degraded into L-proline by microorganisms (such as Bacillus and Actinomycetes) in soil. The degradation half-life is short (≤7 days) and there is no risk of residue. 4. It has good water solubility and can be prepared into aqueous solutions without the addition of solubilizers such as ethanol and Tween, thus reducing application costs and crop damage risks.
[0004] Currently, there is limited research on ester derivatives with L-proline as the lead structure in the field of plant growth regulation. There are no reports on "triethanolamine tri-L-proline ester" and its bioactivity. Developing such compounds is of great significance for filling the gap in green growth regulator technology and promoting green agricultural development. Summary of the Invention
[0005] To address the problems of existing activators such as "low regulation efficiency, high cost, and residual risk", the purpose of this invention is to synthesize triethanolamine tri-L-proline ester through a one-step esterification reaction using L-proline as a raw material. By superimposing the effects of three L-proline molecules, the regulation efficiency can be synergistically improved.
[0006] Another objective of this invention is to provide a simple method for synthesizing the above-mentioned compounds, as well as their application parameters in crops such as wheat and soybeans, so as to provide technical support for the development of green agrochemical products.
[0007] A bioactivator with a trisubstituted L-proline structure, the structural formula of which is: ; The synthesis method of bioactivators with trisubstituted L-proline structures is shown in the following reaction formula: ; This is achieved through the following preparation steps: Triethanolamine and L-proline were mixed and dissolved in toluene. Then, the catalyst was added to the reaction system, and the mixture was heated to 120 °C for 6 h. Water produced in the reaction system was separated using 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, triethanolamine tri-L-proline ester.
[0008] The catalyst is titanium oxysulfate, and the molar ratio of triethanolamine, L-proline and catalyst is 1:(2.5-3.5):(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 with trisubstituted L-proline structures allows them to be used as active or activating ingredients in the regulation of plant growth activity.
[0011] Preferably, it is used as a growth activator for wheat and soybeans.
[0012] Preferably, when used as an activator, the appropriate concentration for promoting the germination and growth of wheat and soybeans is 1-60 ppm.
[0013] Compared with the prior art, the present invention has the following beneficial effects: 1. Innovative structural design: For the first time, the natural branched alkyl group of L-proline is combined with the core active group triethanolamine to form a "three-active-site" molecule, which enhances molecular penetration efficiency and target binding specificity, and significantly improves the regulation efficiency compared with single-structure regulators.
[0014] 2. Process innovation: The synthesis is completed in one step of esterification reaction, without the need for complicated modification steps, with short reaction time, solvent recovery rate ≥90%, and low energy consumption for industrial production.
[0015] 3. Application advantages: good water solubility (35g / L at 25℃), no soil residue (degradation rate ≥98%, 7 days), wide applicability to staple crops, and balances regulatory effect with environmental safety. Detailed Implementation
[0016] 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
[0017] This embodiment describes a method for synthesizing a bioactivator with a trisubstituted L-proline structure, achieved through the following preparation steps: Triethanolamine (0.4 mol), L-proline (1.2 mol), toluene (100 mL), and the catalyst titanium oxysulfate (0.02 mol, 0.05 eq) were added to a 250 mL two-necked flask, and the mixture was heated to 120 °C and reacted for 6 h. Water produced in the reaction system was separated using a water separator. After the reaction was completed, the heating was turned off, and the system was allowed to cool 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 (n-hexane / ethyl acetate = 1:1) to obtain the target product, triethanolamine tri-L-proline ester, in 95% yield.
[0018] Triethanolamine tri-L-proline ester: 1 H NMR (400 MHz, D2O) d 4.61-4.50 (m, 6H), 4.45(t, J= 4.2 Hz, 3H), 3.70(t, J= 4.2 Hz, 6H), 3.35-3.19(m, 6H), 2.42-2.26(m,3H), 2.15-2.01(m, 3H), 2.00-1.88(m, 6H). 13 C NMR (400 MHz, D2O) 173.5, 60.6,58.7, 55.6, 48.2, 28.7, 26.8. HRMS (ESI): [M+H] + calcd for 441.2635, found441.2636. Example 2
[0019] This embodiment describes a method for synthesizing a bioactivator with a substituted L-proline structure. The method involves the following steps: Triethanolamine (0.4 mol), L-proline (1.0 mol), toluene (100 mL), and the catalyst titanium oxysulfate (0.016 mol, 0.04 eq) are added to a 250 mL two-necked flask. The mixture is then heated to 110 °C and reacted for 8 h. Water produced in the reaction system is separated using a water separator. After the reaction is complete, heating is stopped, 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, triethanolamine tri-L-proline ester, in 94% yield. Example 3
[0020] This embodiment describes a method for synthesizing a bioactivator with a trisubstituted L-proline structure, achieved through the following preparation steps: Triethanolamine (0.4 mol), L-proline (1.4 mol), toluene (100 mL), and the catalyst titanium oxysulfate (0.024 mol, 0.06 eq) were added to a 250 mL two-necked flask, and the mixture was heated to 130 °C and reacted for 4 h. Water produced in the reaction system was separated using a water separator. After the reaction was complete, the heating was turned off, and the system was allowed to cool 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 (n-hexane / ethyl acetate = 1:1) to obtain the target product, triethanolamine tri-L-proline ester, in 95% yield.
[0021] Effect Examples Bioactivity Test
[0022] a. Preparation of triethanolamine tri-L-proline ester activator: 0.5 g of triethanolamine tri-L-proline ester (prepared in Example 1) 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, respectively. Tap water treatment served as a control (CK).
[0023] b. 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.
[0024] c. 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.
[0025] Plant height: The standard measurement is the length from the highest point of the leaf to the hypocotyl.
[0026] Root length: The measurement method is to select the three longest roots from a seedling and take their average value.
[0027] 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.
[0028] The results of the bioactivity assays for triethanolamine tri-L-proline ester are shown in Tables 1 and 2. ;
[0029] Bioactivity experiments were conducted on wheat and soybeans, 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 1-60 ppm, and its effect on promoting germination and growth was far superior to the blank control and DA-6 group.
[0030] 2. Using the "soaking method" to test soybean germination and growth, it was found that the compound had the best effect on promoting soybean growth at 1-60 ppm, and its effect on promoting germination and growth was far superior to the blank control and DA-6 group.
[0031] This invention synthesizes triethanolamine tri-L-proline ester using L-proline as a raw material. The process is simple and low-cost, offering the triple advantages of "highly efficient regulation, good water solubility, and no residue." In staple crops such as wheat and soybeans, its regulatory activity is significantly superior to the existing commercial regulator DA-6, with lower effective concentrations and higher environmental safety. Furthermore, the compound's preparation process is energy-efficient, the solvent is recyclable, and it is easily scalable for industrial production. In summary, this invention provides a new direction for the development of green and environmentally friendly activators, possessing significant application value and market potential in improving agricultural product yield and quality and promoting green and sustainable agricultural development.
[0032] 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 with a trisubstituted L-proline structure, characterized in that, Its structural formula is: 。 2. The method for synthesizing the bioactivator with a trisubstituted L-proline structure according to claim 1, characterized in that, This is achieved through the following preparation steps: Triethanolamine and L-proline were mixed and dissolved in toluene. Then, the catalyst was added to the reaction system, and the mixture was heated to 120 °C for 6 h. Water produced in the reaction system was separated using 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, triethanolamine tri-L-proline ester.
3. The method for synthesizing a bioactivator with a trisubstituted L-proline structure according to claim 2, characterized in that, The catalyst is titanium oxysulfate, and the molar ratio of triethanolamine, L-proline and the catalyst is 1:(2.5-3.5):(0.04-0.06).
4. The method for synthesizing a bioactivator with a trisubstituted L-proline structure according to claim 2, characterized in that, The heating reaction is carried out at a temperature of 110-130℃ for 4-8 hours.
5. The application of the bioactivator with the trisubstituted L-proline structure according to claim 1, characterized in that, It can be used as an active ingredient or activator to regulate plant growth activity.
6. The application of the bioactivator with a trisubstituted L-proline structure according to claim 5, characterized in that, It is used as a growth activator for wheat and soybeans.
7. The application of the bioactivator with a trisubstituted L-proline structure according to claim 5, characterized in that: When used as an activator, the appropriate concentration for promoting the germination and growth of wheat and soybeans is 1-60 ppm.