Choline amino acid polyion liquid antibacterial agent and preparation method thereof
By preparing choline amino acid polyionic liquids, the problem of traditional antibacterial drugs easily leading to drug resistance and high toxicity has been solved, and a high-efficiency, low-toxicity, green and environmentally friendly antibacterial material with good biocompatibility and mechanical properties has been achieved.
Patent Information
- Application Number
- CN202410341808.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-03-25
- Publication Date
- 2025-09-26
AI Technical Summary
Existing antibacterial drugs are prone to cause bacterial resistance, and traditional imidazole and pyridyl ionic liquids are highly toxic and cumbersome to prepare, which limits their application in the antibacterial field.
Choline amino acid polyionic liquids were prepared by a simple neutralization reaction using choline-amino acid ionic liquids. The reaction between choline and amino acids was utilized, and the preparation process was green and simple. High-purity P[Cho][AA]PILs-1 and P[Cho][AA]PILs-2 were obtained, which destroyed bacterial cell membranes through electrostatic interactions and hydrophobic insertion.
The prepared choline amino acid polyionic liquid has high antibacterial efficiency, anti-drug resistance, good biocompatibility and mechanical properties, reduced toxicity and improved biodegradability.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of antibacterial materials, and particularly relates to a choline amino acid polyionic liquid antibacterial agent and a preparation method thereof. Background Art
[0002] Bacterial infections pose a serious threat to global public health and safety. Since their introduction, antimicrobial drugs have been widely used. While inhibiting bacteria, they have also led to the development of resistance to traditional drugs. Overuse of antibiotics has also stimulated the evolution of multidrug-resistant microorganisms. There is an urgent need to develop new antimicrobial agents that are highly effective, low-toxic, and less susceptible to drug resistance.
[0003] The unique antimicrobial properties of ionic liquids (ILs) offer new avenues for overcoming current challenges associated with antibiotic-resistant pathogens. Compared to traditional small-molecule antibiotics, ILs and their polymers (polyionic liquids, PILs) mediate irreversible physical disruption of microbial cell membranes, thereby reducing the likelihood of pathogens developing drug resistance and offering significant potential for combating multidrug-resistant microorganisms. The development of (P)ILs as antimicrobial materials currently focuses on traditional ILs such as imidazoles and pyridines. However, the high toxicity, poor biodegradability, and cumbersome preparation processes of imidazole and pyridine-based ILs limit their application in antimicrobial applications. Choline-amino acid ionic liquids ([Cho][AA]ILs) are a class of ionic liquids with excellent biocompatibility. Amino acids (AA) are the building blocks of proteins, while choline (Cho) is a naturally nontoxic cationic substance and an essential nutrient for many mammals. [Cho][AA]ILs can be prepared through the neutralization reaction of choline and amino acids. Only water is introduced as a solvent during the synthesis process. The process is green and simple, and high-purity ILs can be produced without complex purification. In addition, since both anions and cations are derived from non-toxic natural raw materials, the safety of ILs can be improved, biodegradability can be increased, and toxicity can be reduced. Therefore, they have broad development prospects as antibacterial materials.
[0004] Polyionic liquids (PILs) are new materials that use ionic liquids as repeating units and have both the functions of ionic liquids and the excellent mechanical properties of polymers.
[0005] The antimicrobial mechanism of cationic polymers is that the cationic groups generate electrostatic interactions with the negatively charged bacterial surface and insert into the cell membrane through the hydrophobic groups, disrupting the membrane's integrity and leading to indirect leakage of cytoplasmic substances and cell lysis. Compared with the cationic moieties covalently bound to the polymer backbone, the relatively free cations in anionic polymers interact more readily with the negatively charged cell membrane. The free choline cations in PILs can interact with microbial cell membranes through electrostatic attraction. Subsequently, the hydrophobic segments of the PILs insert into the hydrophobic regions of the microbial lipid membrane, ultimately leading to cytoplasmic leakage and cell death. In addition to electrostatic interactions, the hydrophobic chains of the polymers play an important role in interacting with the hydrophobic regions within the cell membrane phospholipid bilayer. Therefore, greater hydrophobicity can lead to better antimicrobial efficacy.
[0006] Choline-amino acid polyionic liquid prepared by polymerization reaction of choline-amino acid ionic liquid has the characteristics of high efficiency and broad-spectrum antibacterial activity, excellent mechanical properties and weak toxicity. It is a green, environmentally friendly, high-efficiency, low-toxic and non-resistant antibacterial material. Summary of the Invention
[0007] The main purpose of the present invention is to provide a new type of choline amino acid polyionic liquid antibacterial agent and its preparation method. The choline amino acid polyionic liquid has high antibacterial properties, anti-drug resistance, and good biocompatibility. The preparation method is simple, green, and environmentally friendly.
[0008] The present invention is achieved through the following technical solutions:
[0009] In the first aspect of the present invention, a double bond is introduced into the amino acid portion to prepare the choline amino acid polyionic liquid P[Cho][AA]PILs-1; in the other aspect, a double bond is introduced into the choline portion to prepare the choline amino acid polyionic liquid P[Cho][AA]PILs-2.
[0010] The preparation method comprises the following steps:
[0011] Step 1: Synthesis of choline amino acid ionic liquid [Cho][AA]ILs: Dissolve the amino acid in deionized water, then add choline hydroxide aqueous solution dropwise, stir the reaction in an ice-water bath, and finally rotary evaporate to obtain pure choline amino acid ionic liquid [Cho][AA]ILs.
[0012] Step 2: Synthesis of Vinylcholine Amino Acid Ionic Liquid V[Cho][AA]VILs-1: Weigh acrylic acid and dissolve it in deionized water. Add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC.HCl) and N-hydroxysuccinimide (NHS). Adjust the pH with sodium hydroxide solution, and add the [Cho][AA]ILs prepared in Step 1. Add EDC again, and rotary evaporation is performed again to obtain the vinylcholine amino acid ionic liquid monomer V[Cho][AA]VILs-1.
[0013] Synthesis of vinyl choline amino acid ionic liquid V[Cho][AA]VILs-2: Weigh acrylic acid, heat it in a water bath, add [Cho][AA]ILs prepared in step 1, and stir thoroughly to obtain the vinyl choline amino acid ionic liquid monomer V[Cho][AA]VILs-2.
[0014] Step 3: Synthesis of choline amino acid polyionic liquid P[Cho][AA]PILs: Dissolve the V[Cho][AA]VILs obtained in the reaction of step 2 in deionized water, stir magnetically until the V[Cho][AA]VILs are completely dissolved, add ammonium persulfate (APS) aqueous solution and N,N,N′,N′-tetramethylethylenediamine (TEMED), place in a hot water bath and stir, and finally dry in a vacuum drying oven to obtain choline amino acid polyionic liquid P[Cho][AA]PILs.
[0015] The difference between P[Cho][AA]PILs-1 and P[Cho][AA]PILs-2 prepared by the present invention lies in the different preparation methods in step 2.
[0016] The amino acids described in the present invention are divided into two types: L-type and D-type. Choline and amino acids react at a ratio of 1:1.01 (mol / mol).
[0017] The synthetic route of P[Cho][AA]PILs-1 prepared by the present invention is as follows Figure 1 As shown, the synthetic route of the prepared P[Cho][AA]PILs-2 is as follows Figure 2 shown.
[0018] In the above technical solution, in step 1, the choline hydroxide aqueous solution is slowly added to the amino acid aqueous solution at a rate of 2 to 3 drops per second using a constant pressure dropping funnel, the ice water bath temperature is 0 to 5° C., and the reaction time is 36 to 72 hours.
[0019] In the above technical solution, the water bath temperature of the rotary evaporator in step 1 is 50-60° C., and the rotation speed is 110-130 r / min.
[0020] In the above technical solution, in the step 2 for preparing V[Cho][AA]VILs-1, the pH value is adjusted to 7-9 with sodium hydroxide solution, the molar ratio of choline amino acid ionic liquid and acrylic acid is 1:1-1:2, the molar ratio of choline amino acid ionic liquid to EDC is 1:1-1:1.5, and the molar ratio of choline amino acid ionic liquid to NHS is 1:0.2-1:0.5.
[0021] In the above technical solution, in step 2, choline-amino acid ionic liquid is added to prepare V[Cho][AA]VILs-1 and stirred for 0.5 to 1 hour. EDC is added once every 3 to 4 hours for a total of 5 to 7 times. After the addition of EDC is completed, the reaction is continued for 10 to 12 hours.
[0022] In the above technical solution, in step 2 for preparing V[Cho][AA]VILs-1, the water bath temperature of the rotary evaporator is 50-60°C and the rotation speed is 110-140 r / min.
[0023] In the above technical solution, in the step 2 for preparing V[Cho][AA]VILs-2, the molar ratio of choline amino acid ionic liquid to acrylic acid is 1:1 to 1:2, the reaction temperature is 50 to 60°C, and the reaction time is 4 to 6 hours.
[0024] In the above technical solution, the amount of APS aqueous solution (0.5-1.0 wt%) added in step 3 is 90-110 μL, and the amount of TEMED is 20-30 μL.
[0025] In the above technical solution, the polymerization temperature in step 3 is 50-60° C., the polymerization time is 4-6 hours, the vacuum drying oven temperature is 40-50° C., and the drying time is 4-6 hours.
[0026] Compared with the prior art, the present invention has the following beneficial effects:
[0027] (1) The polyionic liquid prepared by the present invention is made from natural compounds amino acids and choline. Only water is introduced as a solvent during the reaction process. The process is green and simple. High-purity polyionic liquid can be obtained without complex purification. This can improve the safety of the ionic liquid, increase its biodegradability and reduce its toxicity.
[0028] (2) The choline amino acid polyionic liquid designed and synthesized by the present invention can not only solve the problems of low efficiency, high energy consumption, serious secondary pollution, and strong corrosion to equipment in the traditional ionic liquid esterification reaction, but also can well overcome the problems faced by ionic liquids such as difficult recovery and easy loss;
[0029] (3) The choline amino acid polyionic liquid designed and synthesized in the present invention not only has the high stability and high conductivity of conventional ionic liquids, but also has some unique advantages, such as high antibacterial properties, anti-drug resistance and excellent biocompatibility. BRIEF DESCRIPTION OF THE DRAWINGS
[0030] Figure 1 This is the synthetic route of choline-amino acid polyionic liquid P[Cho][AA]PILs-1.
[0031] Figure 2 This is the synthetic route of choline-amino acid polyionic liquid P[Cho][AA]PILs-2. DETAILED DESCRIPTION
[0032] The present invention will be described in further detail below in conjunction with specific embodiments, which are intended to explain the present invention rather than to limit it. The raw materials to which the present invention relates can be directly purchased from the market. For process parameters not specifically noted, conventional techniques may be used.
[0033] Example 1
[0034] Preparation of choline phenylalanine polyionic liquid LP[Cho][Phe]PILs-1:
[0035] (1) Weigh 1.67 g of L-phenylalanine (Phe) and place it in a round-bottom flask. Add 5 mL of deionized water and stir until the L-phenylalanine is completely dissolved. Then weigh 2.69 g of choline hydroxide aqueous solution and add it dropwise to the L-phenylalanine aqueous solution. The mixture is reacted in an ice-water bath at 3°C for 48 h. After the reaction is complete, the mixture is evaporated using a rotary evaporator. When no water is evaporated from the liquid collection bottle, the evaporation is complete, and the pure choline phenylalanine ionic liquid L-[Cho][Phe]ILs is obtained.
[0036] (2) Weigh 0.72 g of acrylic acid and 5 mL of deionized water into a beaker and mix them evenly with a magnetic stirrer. Add 0.1278 g of EDC and 0.2302 g of NHS and react for 3 h. Adjust the pH value of the resulting solution to 7 with a prepared sodium hydroxide solution. Add 2.68 g of L-[Cho][Phe]ILs and continue stirring. Add 0.1278 g of EDC every 4 h for a total of 5 times. After the addition of EDC is completed, continue the reaction for 12 h. Rotary evaporate the reaction solution twice using a rotary evaporator until no water drops to obtain the vinyl choline-phenylalanine ionic liquid monomer LV[Cho][[Phe]VILs-1.
[0037] (3) The prepared LV[Cho][Phe]VILs-1 was placed in a beaker, 5 mL of deionized water was added, and the mixture was magnetically stirred until completely dissolved. Then, 100 μL of APS aqueous solution (0.8 wt%) and 25 μL of TEMED were added to the solution. The beaker was placed in a 50°C water bath and stirred for 4 h. Finally, the resulting solution was dried in a 40°C vacuum drying oven for 6 h. The resulting product was choline phenylalanine polyionic liquid LP[Cho][Phe]PILs-1.
[0038] Example 2
[0039] Preparation of choline-tryptophan polyionic liquid LP[Cho][Trp]PILs-2:
[0040] (1) Weigh 2.06 g of L-tryptophan (Trp) into a round-bottom flask, add 5 mL of deionized water, and stir until the L-tryptophan is completely dissolved. Then weigh 2.69 g of choline hydroxide aqueous solution and add it dropwise to the L-tryptophan aqueous solution. The mixture is reacted in an ice-water bath at 3°C for 48 h. After the reaction is complete, the mixture is evaporated using a rotary evaporator. When no water is evaporated from the liquid collection bottle, the evaporation is complete, and the pure choline tryptophan ionic liquid L-[Cho][Trp]ILs is obtained.
[0041] (2) Weigh 0.72 g of acrylic acid and add it to a round-bottom flask, which is placed in a 50°C water bath. Slowly add 3.07 g of L-[Cho][Trp]ILs to the round-bottom flask and stir magnetically for 5 h to obtain vinylcholine tryptophan ionic liquid LV[Cho][Trp]VILs-2.
[0042] (3) The prepared LV[Cho][Trp]VILs-2 was placed in a beaker, 5 mL of deionized water was added, and the mixture was magnetically stirred until completely dissolved. Then, 100 μL of APS aqueous solution (0.8 wt%) and 25 μL of TEMED were added to the solution. The beaker was placed in a 50°C water bath and stirred for 4 h. Finally, the resulting solution was dried in a vacuum drying oven at 40°C for 6 h. The resulting product was the choline-tryptophan polyionic liquid LP[Cho][Trp]PILs-2.
[0043] Example 3
[0044] Preparation of choline arginine polyionic liquid LP[Cho][Arg]PILs-2:
[0045] (1) Weigh 1.76 g of L-arginine (Arg) and place it in a round-bottom flask. Add 5 mL of deionized water and stir until the L-arginine is completely dissolved. Then weigh 2.69 g of choline hydroxide aqueous solution and add it dropwise to the L-arginine aqueous solution. The mixture is reacted in an ice-water bath at 3°C for 48 h. After the reaction is complete, use a rotary evaporator to evaporate the mixture until no water is evaporated from the liquid collection bottle. The pure choline arginine ionic liquid L-[Cho][Arg]ILs is obtained.
[0046] (2) Weigh 0.72 g of acrylic acid and add it to a round-bottom flask, which is placed in a 50°C water bath. Slowly add 2.77 g of L-[Cho][Arg]ILs to the round-bottom flask and stir magnetically for 5 h to obtain vinylcholine arginine ionic liquid LV[Cho][Arg]VILs-2.
[0047] (3) The prepared LV[Cho][Arg]VILs-2 was placed in a beaker, 5 mL of deionized water was added, and the mixture was magnetically stirred until completely dissolved. Then, 100 μL of APS aqueous solution (0.8 wt%) and 25 μL of TEMED were added to the solution. The beaker was placed in a 50°C water bath and stirred for 4 h. Finally, the resulting solution was dried in a vacuum drying oven at 40°C for 6 h. The resulting product was the choline arginine polyionic liquid LP[Cho][Arg]PILs-2.
[0048] Example 4
[0049] Preparation of choline arginine polyionic liquid DP[Cho][Arg]PILs-1:
[0050] (1) Weigh 1.76 g of D-arginine (Arg) and place it in a round-bottom flask. Add 5 mL of deionized water and stir until the D-arginine is completely dissolved. Then weigh 2.69 g of choline hydroxide aqueous solution and add it dropwise to the D-arginine aqueous solution. The mixture is reacted in an ice-water bath at 3°C for 48 h. After the reaction is complete, use a rotary evaporator to evaporate the solution until no water is evaporated from the liquid collection bottle. The pure choline arginine ionic liquid D-[Cho][Arg]ILs is obtained.
[0051] (2) Weigh 0.72 g of acrylic acid and 5 mL of deionized water into a beaker and mix them evenly with a magnetic stirrer. Add 0.1278 g of EDC and 0.2302 g of NHS and react for 3 h. Adjust the pH value of the resulting solution to 7 with a prepared sodium hydroxide solution. Add 2.77 g of D-[Cho][Arg]ILs and continue stirring. Add 0.1278 g of EDC every 4 h for a total of 5 times. After the addition of EDC is complete, continue the reaction for 12 h. Rotary evaporate the reaction solution twice using a rotary evaporator until no more water drips to obtain the vinylcholine arginine ionic liquid monomer DV[Cho][Arg]VILs-1.
[0052] (3) The prepared DV[Cho][Arg]VILs-1 was placed in a beaker, 5 mL of deionized water was added, and the mixture was magnetically stirred until completely dissolved. Then, 100 μL of APS aqueous solution (0.8 wt%) and 25 μL of TEMED were added to the solution. The beaker was placed in a 50°C water bath and stirred for 4 h. Finally, the resulting solution was dried in a vacuum drying oven at 40°C for 6 h. The resulting product was choline arginine polyionic liquid DP[Cho][Arg]PILs-1.
[0053] Example 5
[0054] Preparation of choline-tryptophan polyionic liquid DP[Cho][Trp]PILs-2:
[0055] (1) Weigh 2.06 g of D-tryptophan (Trp) into a round-bottom flask, add 5 mL of deionized water, and stir until the D-tryptophan is completely dissolved. Then weigh 2.69 g of choline hydroxide aqueous solution and add it dropwise to the tryptophan aqueous solution. The mixture is reacted in an ice-water bath at 3°C for 48 h. After the reaction is complete, the mixture is evaporated using a rotary evaporator. When no water is evaporated from the liquid collection bottle, the evaporation is complete, and the pure choline tryptophan ionic liquid D-[Cho][Trp]ILs is obtained.
[0056] (2) Weigh 0.72 g of acrylic acid and add it to a round-bottom flask, which is placed in a 50°C water bath. Slowly add 3.07 g of D-[Cho][Trp]ILs to the round-bottom flask and stir magnetically for 5 h to obtain vinylcholine tryptophan ionic liquid DV[Cho][Trp]VILs-2.
[0057] (3) The prepared DV[Cho][Trp]VILs-2 was placed in a beaker, 5 mL of deionized water was added, and the mixture was magnetically stirred until completely dissolved. Then, 100 μL of APS aqueous solution (0.8 wt%) and 25 μL of TEMED were added to the solution. The beaker was placed in a 50°C water bath and stirred for 4 h. Finally, the resulting solution was dried in a vacuum drying oven at 40°C for 6 h. The resulting product was the choline-tryptophan polyionic liquid DP[Cho][Trp]PILs-2.
Claims
1. A choline amino acid polyionic liquid, characterized in that: The polyionic liquid preparation process is divided into two methods: (1) introducing a double bond in the amino acid part and preparing the choline amino acid polyionic liquid P[Cho][AA]PILs-1 through free radical polymerization; (2) introducing a double bond in the choline part and preparing the choline amino acid polyionic liquid P[Cho][AA]PILs-2 through free radical polymerization.
2. The method for preparing the choline amino acid polyionic liquid according to claim 1, comprising the following steps: Step 1: Synthesis of choline amino acid ionic liquid [Cho][AA]ILs: Dissolve the amino acid in deionized water, then add choline hydroxide aqueous solution dropwise, stir the reaction in an ice-water bath, and finally rotary evaporate to obtain pure choline amino acid ionic liquid [Cho][AA]ILs; Step 2: Synthesis of vinylcholine amino acid ionic liquid V[Cho][AA]VILs-1: Weigh acrylic acid and dissolve it in deionized water. Add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl) and N-hydroxysuccinimide (NHS). Adjust the pH with sodium hydroxide solution and add the [Cho][AA]ILs prepared in Step 1. Add EDC again and perform a second rotary evaporation to obtain the vinylcholine amino acid ionic liquid monomer V[Cho][AA]VILs-1. Synthesis of vinyl choline amino acid ionic liquid V[Cho][AA]VILs-2: Weigh acrylic acid, heat it in a water bath, add [Cho][AA]ILs prepared in step 1, and stir thoroughly to obtain the vinyl choline amino acid ionic liquid monomer V[Cho][AA]VILs-2; Step 3: Synthesis of choline amino acid polyionic liquid P[Cho][AA]PILs: Dissolve the V[Cho][AA]VILs obtained in the reaction of step 2 in deionized water, stir magnetically until the V[Cho][AA]VILs are completely dissolved, add ammonium persulfate (APS) aqueous solution and N,N,N′,N′-tetramethylethylenediamine (TEMED), place in a hot water bath and stir, and finally dry in a vacuum drying oven to obtain choline amino acid polyionic liquid P[Cho][AA]PILs.
3. The method for preparing the choline amino acid polyionic liquid according to claim 1, wherein In step 1, the temperature of the ice-water bath is 0-5°C, the reaction time is 36-72 hours, and the water bath temperature of the rotary evaporator is 50-60°C.
4. The method for preparing the choline amino acid polyionic liquid according to claim 1, wherein In the step 2 for preparing V[Cho][AA]VILs-1, the pH value is adjusted to 7-9 with a sodium hydroxide solution, the molar ratio of the choline amino acid ionic liquid to acrylic acid is 1:1-1:2, the molar ratio of the choline amino acid ionic liquid to EDC is 1:1-1:1.5, and the molar ratio of the choline amino acid ionic liquid to NHS is 1:0.2-1:0.
5.
5. The method for preparing the choline amino acid polyionic liquid according to claim 1, wherein In the step 2 for preparing V[Cho][AA]VILs-2, the molar ratio of choline amino acid ionic liquid to acrylic acid is 1:1 to 1:2, the reaction temperature is 50° C. to 60° C., and the reaction time is 4 to 6 hours.
6. The method for preparing the choline amino acid polyionic liquid according to claim 1, wherein In the step 3, the concentration of the APS aqueous solution added is 0.5-1.0 wt %, the polymerization temperature is 50° C.-60° C., and the polymerization time is 4-6 h.