Supramolecular nanohydrogel, preparation method and application thereof

Abstract

The present invention provides a supramolecular nanohydrogel, a preparation method, and an application thereof. The supramolecular nanohydrogel is formed by polymerizing a modified supramolecular monomer and a functional monomer under the action of a crosslinker. The modified supramolecular monomer selected by the present invention is based on a 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)urea group, and a long alkane chain containing a carbon-carbon double bond is introduced into its molecular structure to impart stronger hydrophobicity and enhance non-covalent interaction, thereby enabling complementation with the hydrophobic domain of the Art v1 protein. Furthermore, both the modified supramolecular monomer and the functional monomer contain double bonds and can undergo free radical polymerization to form the supramolecular nanohydrogel. The functional groups on the functional monomer are integrated into the supramolecular nanohydrogel through a polymerization reaction, so that the newly formed supramolecular nanopolymer binds to the target protein through non-covalent interactions such as electrostatic, hydrophobic, and hydrogen bonds, thereby specifically binding to the Artemisia pollen allergen protein Art v1.

Description

Technical Field

[0001] The present invention relates to the technical field of supramolecular polymers and preparation thereof, in particular to a supramolecular nano hydrogel, a preparation method and application thereof. Background Art

[0002] Pollen allergy is an immune system disorder with the primary symptoms being sneezing (96.78%) and nasal itching (91.6%). Itchy eyes are particularly prominent in 68.58% of cases. Other symptoms include fatigue (31.07%) and lethargy (30.49%). If left untreated, severe cases can develop asthma and shock. Studies have shown that Artemisia pollen is a major source of pollen allergy. In Europe, 10%-14% of hay fever sufferers experience an allergic reaction to Artemisia pollen. In China, pollen from Artemisia plants is a significant allergenic factor in northern China, with the highest allergy rate occurring between August and October. The rate of Artemisia pollen allergy in northern China is 24.1%. The allergenic proteins in Artemisia are Artv1, Artv2, Artv3, Artv4, Artv5, and Artv6. Further characterization has revealed that Artv1 is the primary allergen, affecting 95% of patients with Artemisia pollen allergy.

[0003] Currently, for Artemisia pollen allergy, symptoms are relieved by taking general anti-allergic drugs. However, since the drugs are toxic and long-term use can easily lead to drug resistance, the effect is poor. Although specific treatment can solve the problem from the source to a certain extent, patients need to start treatment 4 to 6 months before the pollen season. The current application range is narrow and the price is expensive. The medication cycle is long, and improper dosage control is prone to occur during the medication process, leading to allergic symptoms and reduced patient compliance. Therefore, it is not suitable for most patients.

[0004] Therefore, it is particularly important to develop an Artemisia pollen allergen protein therapeutic agent in the field of biomedicine that is simple to prepare, economical and practical, easy to implement and fast-acting. Summary of the Invention

[0005] In response to the above-mentioned problems existing in the prior art, the present invention provides a supramolecular nanohydrogel and its preparation method and application, so as to provide a specific adsorption material for the Artemisia pollen allergen protein Art v1, thereby solving the problems of high cost, long medication cycle and unstable effect of Artemisia pollen allergy treatment.

[0006] The specific content of the invention is as follows:

[0007] In a first aspect, the present invention provides a supramolecular nanohydrogel microsphere, wherein the supramolecular nanohydrogel is formed by polymerization of 10-55% modified supramolecular monomer, 35-89% functional monomer and 1-10% crosslinking agent, based on a molar weight of 100; wherein,

[0008] The functional monomers include one or more of N-isopropylacrylamide, N-tert-butylacrylamide, N-hydroxymethylacrylamide, N-hydroxyethylacrylamide, methacrylic acid, acrylic acid, crotonic acid, 2-chloroacrylic acid and 2-bromoacrylic acid;

[0009] The modified supramolecular monomers include: 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]butanecarbamoyloxy}hydroxypropyl acrylamide, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]hexanecarbamoyloxy}hydroxyethyl methacrylate, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]octanecarbamoyloxy}hydroxyethyl methacrylate, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]octanecarbamoyloxy}hydroxyethyl methacrylate, one or more of 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]hexanecarbamoyloxy}hydroxyethyl acrylamide, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]butanecarbamoyloxy}butyl acrylate, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]hexanecarbamoyloxy}butyl acrylate and 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]octanecarbamoyloxy}methyl acrylate

[0010] The cross-linking agent is N,N-methylenebisacrylamide.

[0011] Optionally, the functional monomer is one or more of N-hydroxymethyl acrylamide, crotonic acid, methacrylic acid, 2-bromoacrylic acid, acrylic acid and 2-chloroacrylic acid.

[0012] Optionally, the modified supramolecular monomer is one or more of 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]butanecarbamoyloxy}hydroxypropyl acrylamide, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]octanecarbamoyloxy}methyl acrylate and 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]hexanecarbamoyloxy}hydroxypropyl acrylamide.

[0013] Optionally, the supramolecular nanohydrogel is a nanosphere with a particle size of 30 to 150 nm.

[0014] In a second aspect, the present invention provides a method for preparing the supramolecular nanohydrogel described in the first aspect, wherein the supramolecular nanohydrogel microspheres are prepared by the following method:

[0015] S1, dissolving the modified supramolecular monomer in an organic solvent to form a first solution;

[0016] S2, dissolving the functional monomer and the cross-linking agent in ultrapure water to obtain a second solution;

[0017] S3. Under a nitrogen atmosphere, the first solution and the second solution are mixed, and an appropriate amount of initiator and surfactant are added. The resulting mixed system is continuously stirred at 40-95° C. to allow the polymerization reaction to proceed;

[0018] S4. After the polymerization reaction is completed, the reaction system is transferred to a dialysis bag and dialyzed with excess ultrapure water for 1 to 10 days to obtain the supramolecular nanohydrogel.

[0019] Optionally, in S1, the organic solvent is methanol, ethanol, acetonitrile, N,N-dimethylformamide or acetone.

[0020] Optionally, in the mixed system, the molar ratio of the modified supramolecular monomer, the functional monomer and the cross-linking agent is 10-55:35-89:1-10;

[0021] The initiator accounts for 1% to 10% of the total molar amount of the modified supramolecular monomer, functional monomer and cross-linking agent;

[0022] The surfactant accounts for 0.02% to 0.15% of the total molar amount of the modified supramolecular monomer, the functional monomer and the cross-linking agent.

[0023] Optionally, in S3, the initiator is selected from one or more of ammonium persulfate, potassium persulfate, sodium persulfate, azobisisobutyronitrile, azobisisoheptanenitrile, and dimethyl azobisisobutyrate;

[0024] The surfactant is selected from one or more of sodium fatty alcohol polyoxyethylene ether sulfate, sodium lignin sulfonate, sodium lauryl sulfate, sodium dodecylbenzene sulfonate, hexadecyltrimethylammonium bromide, sorbitan monolaurate, polysorbate 80 and lauroyl glutamate.

[0025] Optionally, in S3, the stirring speed is 100 to 500 rpm.

[0026] In a third aspect, the present invention provides a use of the supramolecular nanohydrogel described in the first aspect, wherein the supramolecular nanohydrogel is used as a raw material for preparing a drug for treating wormwood pollen allergy;

[0027] The supramolecular nanohydrogel has a specific adsorption function for Artemisia pollen allergen protein Art v1.

[0028] Compared with the prior art, the present invention has the following advantages:

[0029] The present invention provides a supramolecular nanohydrogel, which is formed by polymerizing modified supramolecular monomers and functional monomers under the action of a crosslinking agent. The modified supramolecular monomer selected by the present invention is based on 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)urea, and a long alkane chain containing a carbon-carbon double bond is introduced into its molecular structure, so that the modified supramolecular monomer has stronger hydrophobicity and enhanced non-covalent interaction, and can complement the hydrophobic domain of the Art v1 protein. In addition, the modified supramolecular monomer and the functional monomer both contain double bonds and can undergo free radical polymerization to form the supramolecular nanohydrogel. The functional groups on the functional monomer are integrated into the supramolecular nanohydrogel through a polymerization reaction, so that the newly formed supramolecular nanopolymer binds to the target protein in the form of non-covalent interactions such as electrostatic, hydrophobic, and hydrogen bonds, thereby having the function of specifically binding to the artemisia pollen allergen protein Art v1.

[0030] The supramolecular nanohydrogel provided by the present invention is simple to prepare, the raw materials are cheap and easily available, and the storage cost is low; the supramolecular nanohydrogel microspheres are used as wormwood pollen protein allergen adsorbents to treat symptoms such as rhinitis caused by wormwood pollen allergy, which can greatly reduce the cost of medication, shorten the medication time, and increase patient compliance. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0032] Figure 1 The flowchart of the preparation method of the supramolecular nanohydrogel provided by the embodiment of the present invention is shown;

[0033] Figure 2 shows a scanning electron microscope image of the supramolecular nanohydrogel provided by an embodiment of the present invention;

[0034] Figure 3 shows the infrared spectrum of the modified supramolecular monomer provided by an embodiment of the present invention;

[0035] Figure 4The figure shows the adsorption of Art v1 protein by the supramolecular nanohydrogel provided in the embodiment of the present invention at different times;

[0036] Figure 5 The figure shows the adsorption of Art v1 protein at different concentrations by the supramolecular nanohydrogel provided in the embodiment of the present invention;

[0037] Figure 6 The graph shows the adsorption of different proteins in plasma by the supramolecular nanohydrogel provided by the embodiment of the present invention. DETAILED DESCRIPTION

[0038] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of the embodiments. The following description of at least one exemplary embodiment is actually only illustrative and is by no means a limitation on the present invention and its application or use. Based on the embodiments of the present invention, any product that is identical or similar to the present invention and is obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts falls within the scope of protection of the present invention. In addition, all other embodiments obtained by ordinary technicians in this field without carrying out creative work fall within the scope of protection of the present invention.

[0039] Where specific experimental steps or conditions are not specified in the examples, the conventional experimental steps or conditions described in the prior art in the art may be used. Reagents and other instruments used, for which the manufacturer is not specified, are commercially available conventional reagent products. Furthermore, the accompanying drawings are merely schematic illustrations of embodiments of the present invention and are not necessarily drawn to scale. Identical reference numerals in the figures denote identical or similar parts, and their repeated descriptions will be omitted. Some block diagrams shown in the accompanying drawings are functional entities and do not necessarily correspond to physically or logically independent entities.

[0040] Technologies, methods, and equipment known to those skilled in the art may not be discussed in detail, but where appropriate, such technologies, methods, and equipment should be considered part of the description of the present invention.

[0041] In the description of the present invention, it should be understood that the use of terms such as "first" and "second" to limit components is only for the convenience of distinguishing the corresponding components. Unless otherwise stated, the above terms have no special meaning and therefore cannot be understood as limiting the scope of protection of the present invention.

[0042] In addition, the technical features involved in the different embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0043] Supramolecular hydrogels are a class of polymeric materials formed by the self-assembly of low-molecular-weight gelling agents through reversible supramolecular non-covalent interactions (such as hydrogen bonding, π-π stacking, van der Waals forces, and host-guest interactions). Due to these reversible non-covalent interactions, the prepared polymeric materials are often stimuli-responsive. By adjusting the structure of the gelling agents or introducing functional groups, the properties of supramolecular hydrogels can be customized to suit specific application requirements. Due to the biocompatibility of hydrogels, supramolecular hydrogels are often designed into nanostructures to meet the requirements of drug loading, targeted delivery, and affinity for biomacromolecules.

[0044] Supramolecular nanohydrogels possess highly tunable network structures and large surface areas, making them ideal materials for capturing and separating biomacromolecules. By introducing specific recognition groups (such as antibodies, ligands, and nucleic acid aptamers) into the nanohydrogels, highly selective capture of target biomacromolecules can be achieved. For example, they can be used to capture specific proteins, enzymes, or DNA fragments.

[0045] The interactions between biomacromolecules are characterized by reversibility, specificity, and multiple sites of action. These interactions primarily rely on intermolecular non-covalent forces—electrostatic forces, hydrophobic forces, hydrogen bonds, and van der Waals forces. This invention utilizes the functional groups of the Artemisia v1 allergen protein to design nanogel macromolecules that interact with them, thereby achieving specific binding between the nanogel macromolecules and the protein. Specific implementations are as follows:

[0046] In a first aspect, the present invention provides a supramolecular nano-hydrogel, wherein the supramolecular nano-hydrogel is formed by polymerization of a modified supramolecular monomer and a functional monomer under the action of a cross-linking agent; wherein,

[0047] The functional monomers include one or more of N-isopropylacrylamide, N-tert-butylacrylamide, N-hydroxymethylacrylamide, N-hydroxyethylacrylamide, methacrylic acid, acrylic acid, crotonic acid, 2-chloroacrylic acid and 2-bromoacrylic acid;

[0048] The cross-linking agent is N,N-methylenebisacrylamide;

[0049] The molar ratio of the modified supramolecular monomer, the functional monomer and the cross-linking agent is 10:89:1 to 55:35:10.

[0050] The embodiment of the present invention uses the supramolecular monomer 6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)urea group that can complement the hydrophobic domain of Art v1 protein to perform a simple modification (introducing a long alkane chain containing a carbon-carbon double bond) to improve its hydrophobicity and enhance the non-covalent interaction force, thereby further enhancing its complementary ability with the hydrophobic domain of Art v1 protein. The modified supramolecular monomer and functional monomer both contain double bonds and can undergo free radical polymerization to form supramolecular nanohydrogel microspheres. The functional groups on the functional monomers are integrated into the supramolecular nanohydrogel microspheres through polymerization reaction, so that the newly formed supramolecular nanopolymer binds to the target protein in the form of non-covalent interactions such as electrostatic, hydrophobic, and hydrogen bonds, so that the modified supramolecular monomer has the ability to specifically bind to the Artemisia pollen allergen protein Art. v1 function; the modified supramolecular monomers selected in the embodiment of the present invention include: 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]butanecarbamoyloxy}hydroxypropyl acrylamide, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]hexanecarbamoyloxy}hydroxyethyl methacrylate, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]octanecarbamoyloxy}hydroxyethyl methacrylate, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]hexanecarbamoyloxy}hydroxyethyl acrylamide, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]butanecarbamoyloxy}acrylamide The modified supramolecular monomer is one or more of butyl acrylate, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl) ureido] hexanecarbamoyloxy} butyl acrylate and 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl) ureido] octanecarbamoyloxy} methyl acrylate, and the preferred modified supramolecular monomer is one or more of 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl) ureido] butanecarbamoyloxy} hydroxypropyl acrylamide, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl) ureido] octanecarbamoyloxy} methyl acrylate and 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl) ureido] hexanecarbamoyloxy} hydroxypropyl acrylamide.

[0051] In specific implementation, the functional monomer used in the embodiment of the present invention also contains a double bond, which can undergo free radical polymerization with the modified supramolecular monomer to form supramolecular nanohydrogel microspheres. The functional groups on the functional monomer are integrated into the supramolecular nanohydrogel through polymerization reaction, so that the newly formed supramolecular nanopolymer binds to the target protein in the form of non-covalent interactions such as electrostatic, hydrophobic, and hydrogen bonds, thereby improving the specific binding ability. The preferred functional monomer is one or more of N-hydroxymethyl acrylamide, crotonic acid, methacrylic acid, 2-bromoacrylic acid, acrylic acid, and 2-chloroacrylic acid.

[0052] In a second aspect, the present invention provides a method for preparing the supramolecular nanohydrogel described in the first aspect. Figure 1 The flow chart of the preparation method of supramolecular nanohydrogel provided by the embodiment of the present invention is shown as follows: Figure 1 As shown, the supramolecular nanohydrogel is prepared by the following method:

[0053] S1, dissolving the modified supramolecular monomer in an organic solvent to form a first solution;

[0054] During this step, the hydrophobically modified supramolecular monomer is insoluble in water. This requires that the selected organic solvent has good solubility for the modified supramolecular monomer, and the organic solvent is selected from methanol, ethanol, acetonitrile, N,N-dimethylformamide, or acetone. Pre-dissolving the modified supramolecular monomer before adding it to the reaction system facilitates the reaction.

[0055] S2, dissolving the functional monomer and the cross-linking agent in ultrapure water to obtain a second solution;

[0056] When this step is specifically implemented, the functional monomer and the cross-linking agent can be fully dissolved in ultrapure water by ultrasonic treatment; the cross-linking agent is selected from N,N-methylenebisacrylamide, and the cross-linking agent can convert linear molecules into three-dimensional molecules to cross-link the polymer chains into supramolecular nanohydrogels.

[0057] S3. Under a nitrogen atmosphere, the first solution and the second solution are mixed, and an appropriate amount of initiator and surfactant are added. The resulting mixed system is continuously stirred at 40-95° C. to allow the polymerization reaction to proceed;

[0058] During the specific implementation of this step, an initiator and a surfactant are added to the polymerization reaction system to improve the reaction conditions and environment. The selected initiator is easily decomposed by heat, is easily soluble in water or ethanol, and can rapidly generate free radicals to initiate the polymerization reaction between the supramolecular monomer and the functional monomer. The selected surfactant can provide interfacial tension, so that the supramolecular nanohydrogel formed during the polymerization process has uniform particle size, good sphericity, and stable properties. The initiator is selected from one or more of ammonium persulfate, potassium persulfate, sodium persulfate, azobisisobutyronitrile, azobisisoheptanonitrile, and dimethyl azobisisobutyrate; the surfactant is selected from one or more of sodium fatty alcohol polyoxyethylene ether sulfate, sodium lignin sulfonate, sodium dodecyl sulfate, sodium dodecylbenzenesulfonate, cetyltrimethylammonium bromide, sorbitan monolaurate, polysorbate 80, and lauroyl glutamate.

[0059] Furthermore, the amounts of initiator and surfactant influence the polymerization reaction rate and extent, thus affecting the particle size, stability, and adsorption properties of the supramolecular nanohydrogel material. In this embodiment, the initiator accounts for 1% to 10% of the total molar weight of the modified supramolecular monomer, functional monomer, and crosslinker, and the surfactant accounts for 0.02% to 0.15% of the total molar weight of the modified supramolecular monomer, functional monomer, and crosslinker. Under these amounts, the synthesized supramolecular nanohydrogel material has an ideal particle size, uniform shape and size, stable properties, and good adsorption of the target protein.

[0060] Furthermore, the reaction temperature is preferably 40-95°C; the stirring speed is preferably 50-500 rpm. The reaction temperature affects the degree of polymerization of free radical polymerization, and the stirring speed affects the size and shape of the polymer. Under the above temperature and stirring speed, supramolecular nano-hydrogels with appropriate size and shape can be successfully prepared.

[0061] S4. After the polymerization reaction is completed, the reaction system is transferred to a dialysis bag and dialyzed with excess ultrapure water for 1 to 10 days to obtain the supramolecular nanohydrogel.

[0062] When this step is specifically implemented, ultrapure water treatment is used to remove impurities in the reaction system to obtain purified supramolecular nanohydrogel.

[0063] It should be noted that the molar ratio of the modified supramolecular monomer, functional monomer, and crosslinker involved in the polymerization reaction is 10-55:35-89:1-10. The amount of crosslinker (N,N-methylenebisacrylamide) affects the morphology and particle size of the resulting supramolecular nanohydrogels: excessive addition tends to oversize the microspheres, while insufficient crosslinking results in low yield. At these dosages, the prepared supramolecular nanohydrogels exhibit uniform particle size and high yield. The crosslinker crosslinks the 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]} group with the functional monomer, creating more interaction sites and increasing the specific binding sites and binding strength with the Art v1 protein.

[0064] In a third aspect, the present invention provides an application of the supramolecular nanohydrogel described in the first aspect, wherein the supramolecular nanohydrogel microspheres are used as a raw material for preparing a drug for treating wormwood pollen allergy;

[0065] The supramolecular nanohydrogel has a specific adsorption function for the Artemisia allergen protein Art v1.

[0066] The supramolecular nanohydrogel provided by the present invention is based on the principle of antigen-antibody interaction. A specific supramolecular monomer (2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)urea) that can bind to the artemisia pollen allergen protein Art v 1 is selected for modification and the supramolecular nanohydrogel is constructed. The obtained supramolecular nanohydrogel has extremely low adsorption effect on other proteins in the blood, such as serum albumin and immunoglobulin, but has specific adsorption effect on the main artemisia pollen allergen protein Art v 1, and the adsorption effect is good.

[0067] In order to enable those skilled in the art to more clearly understand the present invention, the supramolecular nano-hydrogel microspheres, preparation method and application of the present invention are described in detail through the following examples.

[0068] Example 1

[0069] (1) Preparation of supramolecular nanohydrogels

[0070] Dissolving 30 mM of the modified supramolecular monomer 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]butanecarbamoyloxy}hydroxypropylacrylamide in 2 mL of ethanol to obtain a first solution;

[0071] 40 mM N-hydroxymethyl acrylamide, 25 mM crotonic acid, and 5 mM cross-linking agent N,N-methylenebisacrylamide were dissolved in water and then sonicated for 60 minutes to obtain a second solution;

[0072] The first and second solutions were poured into a flask to obtain a mixed solution. Then, sodium persulfate (an initiator) and cetyltrimethylammonium bromide (a surfactant) were added to the solution. The sodium persulfate amount was 5% of the total molar weight of the modified supramolecular monomer, functional monomer, and crosslinker, and the cetyltrimethylammonium bromide amount was 0.13% of the total molar weight of the modified supramolecular monomer, functional monomer, and crosslinker. The reaction was carried out at 90°C, with a stirring speed of 150 rpm and nitrogen flow for 15 hours. The apparatus was then removed, the mixture was allowed to cool naturally, and then transferred to a dialysis bag. The mixture was dialyzed against excess ultrapure water for one day to obtain a supramolecular nanohydrogel with adsorption properties.

[0073] Figure 3 The infrared spectra of the modified supramolecular monomers provided by the embodiments of the present invention are shown, wherein Figure (1) is the infrared spectrum of 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]butanecarbamoyloxy}hydroxypropyl acrylamide, Figure (2) is the infrared spectrum of 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]butanecarbamoyloxy}hydroxypropyl acrylamide, and Figure (3) is the infrared spectrum of 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]octanecarbamoyloxy}methyl acrylate; Figure 3 As shown, 3311, 3319, 3327cm -1 The strong absorption peaks nearby are the NH stretching vibration of -NHCOO-, 1658, 1666, 1669 cm -1 The above results indicate that the modified supramolecular monomer has been successfully synthesized due to the absorption of C=C stretching vibration. Moreover, the compound can generate free radicals and polymerize into nanogels.

[0074] Figure 2 FIG shows a scanning electron microscope image of the supramolecular nanohydrogel provided by an embodiment of the present invention, as shown in FIG. Figure 2 As shown, the supramolecular nano-hydrogel microspheres prepared in this embodiment have moderate particle size, uniform sphere shape, particle size between 30 and 150 nm, and good dispersibility.

[0075] (2) Adsorption kinetics experiment

[0076] Deionized water, Art v1 protein solution, buffer, and supramolecular nanohydrogel were added to a centrifuge tube. Mixed at a constant temperature and oscillation speed for varying times (0.5 min, 1 min, 2 min, 3 min, 5 min, 10 min, 30 min, and 60 min), the mixture was centrifuged, and the supernatant was added to a liquid chromatography vial. The remaining Art v1 protein concentration was determined by HPLC, and the adsorption rate and amount of Art v1 protein by the supramolecular nanohydrogel at different times were calculated.

[0077] Figure 4 The results show the adsorption of Art v1 protein by the supramolecular nanohydrogel provided by the embodiment of the present invention at different times. Figure 4 As shown in the figure, the adsorption of protein Art v1 by supramolecular nanohydrogel increased sharply in the first 10 minutes and reached equilibrium after 10 minutes, with an adsorption rate of 95%.

[0078] (3) Isothermal adsorption experiment

[0079] Deionized water, Art v1 protein solutions of varying concentrations (0.004 mg / mL, 0.006 mg / mL, 0.008 mg / mL, 0.01 mg / mL, 0.02 mg / mL, 0.03 mg / mL, 0.04 mg / mL, 0.05 mg / mL, and 0.06 mg / mL), buffer, and nanohydrogel microspheres were added to a centrifuge tube. Mixing was performed at a constant temperature and oscillation rate for 3 hours, followed by centrifugation. The supernatant was collected and added to a liquid chromatography vial. The remaining Art v1 protein concentration was determined by HPLC, and the amount of Art v1 protein adsorbed at different times was calculated.

[0080] Figure 5 The results show the adsorption of different concentrations of Art v1 protein by the supramolecular nanohydrogel provided in the embodiment of the present invention. Figure 5 As shown in the figure, with the increase of Art v1 protein concentration, the adsorption amount of protein by supramolecular nanopolymers showed a trend of rapid growth-gradual equilibrium. When the protein concentration was 0.04 mg / mL, the adsorption reached saturation, and the maximum adsorption amount was about 0.4533 nmol / mg, which was much higher than the minimum sensitizing amount (0.045 nmol / mg).

[0081] (4) Specific adsorption experiment

[0082] Deionized water, different protein solutions (Art v1 protein solution, immunoglobulin A (IgA), hemoglobin (HGB), β-globulin, immunoglobulin E (IgE) γ-globulin), buffer, and supramolecular nanohydrogel were added to a centrifuge tube. Mixing and shaking were performed at a specific temperature and speed for 240 minutes. The mixture was then centrifuged and the supernatant was collected and added to a liquid chromatography vial. The remaining Art v1 protein concentration was determined by high-performance liquid chromatography, and the adsorption rate of the supramolecular nanohydrogel for the different proteins was calculated.

[0083] Figure 6 The adsorption of different proteins in plasma by the supramolecular nanohydrogel provided in the embodiment of the present invention is shown in FIG. Figure 6As shown, the adsorption rate of supramolecular nanohydrogel to IgA, HGB, β-globulin, IgE, and γ-globulin in plasma is extremely low, less than 5%, and the adsorption rate to Art v1 is 90%, indicating that the adsorption of supramolecular nanohydrogel to Art v1 is specific and has the prospect of being used as an "artificial antibody" for Artemisia pollen allergy.

[0084] Example 2

[0085] Dissolving 15 mM of the modified supramolecular monomer 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]butanecarbamoyloxy}hydroxypropylacrylamide in 2 mL of acetonitrile to obtain a first solution;

[0086] 30 mM methacrylic acid, 53 mM 2-bromoacrylic acid, and 2 mM cross-linking agent N,N-methylenebisacrylamide were dissolved in water and then sonicated for 60 minutes to obtain a second solution;

[0087] The first and second solutions were poured into a flask to obtain a mixed solution. Azobisisobutyronitrile (AIBN) was added as an initiator, and sodium lignin sulfonate (SLS) was added as a surfactant. The amount of AIBN was 1% of the total molar weight of the modified supramolecular monomer, functional monomer, and crosslinker, and the amount of sodium lignin sulfonate was 0.05% of the total molar weight of the modified supramolecular monomer, functional monomer, and crosslinker. The mixture was reacted at 45°C, stirred at 450 rpm, and purged with nitrogen for 5 hours. The mixture was then removed from the apparatus, cooled naturally, and transferred to a dialysis bag. The mixture was dialyzed against excess ultrapure water for 3 days to obtain a supramolecular nanohydrogel with adsorption properties.

[0088] The adsorption rate of the supramolecular nanohydrogel provided in this embodiment for Art v1 protein is 90%.

[0089] Example 3

[0090] 53 mM of the modified supramolecular monomer 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]octanecarbamoyloxy}acrylate was dissolved in 2 mL of dimethyl sulfoxide to obtain a first solution;

[0091] 10 mM acrylic acid, 31 mM 2-chloroacrylic acid, and 6 mM cross-linking agent N,N-methylenebisacrylamide were dissolved in water and then sonicated for 30 minutes to obtain a second solution;

[0092] The first and second solutions were poured into a flask to obtain a mixed solution. The initiator, ammonium persulfate, and the surfactant, sodium dodecylbenzenesulfonate, were then added to the solution. The initiator amounted to 8% of the total molar weight of the modified supramolecular monomer, functional monomer, and crosslinker, and the surfactant amounted to 0.03% of the total molar weight of the modified supramolecular monomer, functional monomer, and crosslinker. The reaction was carried out at 40°C, with a stirring speed of 350 rpm and nitrogen flow for 13 hours. The apparatus was then removed, the mixture was allowed to cool naturally, and then transferred to a dialysis bag. The mixture was dialyzed against excess ultrapure water for 9 days to obtain a supramolecular nanohydrogel with adsorption properties.

[0093] The adsorption rate of the supramolecular nanohydrogel provided in this embodiment for Art v1 protein is 95%.

[0094] In summary, the supramolecular nanohydrogel provided in this embodiment has the advantages of simple preparation, cheap and readily available raw materials, rapid action, and good adsorption effect. It can be used to adsorb the Artemisia pollen allergen protein Art v1 and has the prospect of being used as an "artificial antibody" for the treatment of Artemisia pollen allergy. It can solve the current problems of long treatment time and high cost of Artemisia pollen allergy.

[0095] In the description of this specification, the reference terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" mean that the specific features, structures, materials, or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in any appropriate manner in any one or more embodiments or examples. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification.

[0096] For simplicity of description, the method embodiments are described as a series of actions. However, those skilled in the art should be aware that the present invention is not limited by the order of the actions described, as certain steps can be performed in other orders or simultaneously according to the present invention. Furthermore, those skilled in the art should also be aware that the embodiments described in this specification are preferred embodiments, and the actions and components involved are not necessarily required for the present invention.

[0097] The above is a detailed introduction to a supramolecular nanohydrogel, preparation method and application provided by the present invention. Specific examples are used herein to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only used to help understand the method and core concept of the present invention. At the same time, for those skilled in the art, according to the concept of the present invention, there may be changes in the specific implementation methods and application scopes. In summary, the content of this specification should not be understood as limiting the present invention.

Claims

1. A supramolecular nanohydrogel, characterized in that: The supramolecular nano hydrogel is formed by polymerization of 10-55% modified supramolecular monomers, 35-89% functional monomers and 1-10% crosslinking agents, based on a molar weight of 100%. The functional monomers include one or more of N-isopropylacrylamide, N-tert-butylacrylamide, N-hydroxymethylacrylamide, N-hydroxyethylacrylamide, methacrylic acid, acrylic acid, crotonic acid, 2-chloroacrylic acid and 2-bromoacrylic acid; The modified supramolecular monomers include: 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]butanecarbamoyloxy}hydroxypropyl acrylamide, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]hexanecarbamoyloxy}hydroxyethyl methacrylate, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]octanecarbamoyloxy}hydroxyethyl methacrylate, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]octanecarbamoyloxy}hydroxyethyl methacrylate, one or more of 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]hexanecarbamoyloxy}hydroxyethyl acrylamide, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]butanecarbamoyloxy}butyl acrylate, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]hexanecarbamoyloxy}butyl acrylate and 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]octanecarbamoyloxy}methyl acrylate The cross-linking agent is N,N-methylenebisacrylamide.

2. The supramolecular nano-hydrogel microspheres according to claim 1, characterized in that: The functional monomer is one or more of N-hydroxymethyl acrylamide, crotonic acid, methacrylic acid, 2-bromoacrylic acid, acrylic acid and 2-chloroacrylic acid.

3. The supramolecular nano-hydrogel microspheres according to claim 1, characterized in that: The modified supramolecular monomer is one or more of 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]butanecarbamoyloxy}hydroxypropyl acrylamide, 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]octanecarbamoyloxy}methyl acrylate and 2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]hexanecarbamoyloxy}hydroxypropyl acrylamide.

4. The supramolecular nanohydrogel according to claim 1, characterized in that The supramolecular nano hydrogel is a nano microsphere with a particle size of 30 to 150 nm.

5. A method for preparing the supramolecular nanohydrogel according to any one of claims 1 to 4, characterized in that: The supramolecular nano hydrogel is prepared by the following method: S1, dissolving the modified supramolecular monomer in an organic solvent to form a first solution; S2, dissolving the functional monomer and the cross-linking agent in ultrapure water to obtain a second solution; S3. Under a nitrogen atmosphere, the first solution and the second solution are mixed, and an appropriate amount of initiator and surfactant are added. The resulting mixed system is continuously stirred at 40-95° C. to allow the polymerization reaction to proceed; S4. After the polymerization reaction is completed, the reaction system is transferred to a dialysis bag and dialyzed with excess ultrapure water for 1 to 10 days to obtain the supramolecular nanohydrogel.

6. The method for preparing the supramolecular nanohydrogel according to claim 5, characterized in that: In S1, the organic solvent is methanol, ethanol, acetonitrile, N,N-dimethylformamide or acetone.

7. The method for preparing the supramolecular nanohydrogel according to claim 5, characterized in that: In the mixed system, the molar ratio of the modified supramolecular monomer, the functional monomer and the cross-linking agent is 10-55:35-89:1-10; The initiator accounts for 1% to 10% of the total molar amount of the modified supramolecular monomer, functional monomer and cross-linking agent; The surfactant accounts for 0.02% to 0.15% of the total molar amount of the modified supramolecular monomer, the functional monomer and the cross-linking agent.

8. The method for preparing the supramolecular nanohydrogel according to claim 5, characterized in that: In S3, the initiator is selected from one or more of ammonium persulfate, potassium persulfate, sodium persulfate, azobisisobutyronitrile, azobisisoheptanenitrile, and dimethyl azobisisobutyrate; The surfactant is selected from one or more of sodium fatty alcohol polyoxyethylene ether sulfate, sodium lignin sulfonate, sodium lauryl sulfate, sodium dodecylbenzene sulfonate, hexadecyltrimethylammonium bromide, sorbitan monolaurate, polysorbate 80 and lauroyl glutamate.

9. The method for preparing the supramolecular nanohydrogel according to claim 5, characterized in that: In S3, the stirring speed is 100 to 500 rpm.

10. An application of the supramolecular nanohydrogel according to any one of claims 1 to 4, characterized in that: The supramolecular nano-hydrogel microspheres are used as raw materials for preparing medicines for allergy to wormwood pollen; The supramolecular nanohydrogel has a specific adsorption function for the Artemisia allergen protein Art v1.

Images