A polyethylene glycol tail chain water-soluble polyphenol polymer and its preparation method and application

Abstract

The invention relates to a polyethylene glycol tail chain water-soluble polyphenol polymer and a preparation method and application thereof. Alkynylphenol ligand and methoxy polyethylene glycol azide are used as raw materials. N,N,N',N,'N"-pentamethyldiethylenetriamine is added under nitrogen, cuprous bromide is used as a catalyst, and N,N-dimethylformamide is used as a solvent. After reacting at 60-80 DEG C for 24-48 hours, an EDTA solution is added, ultrafiltration centrifugation is performed, and freeze drying is performed to obtain the polyethylene glycol tail chain water-soluble polyphenol polymer. Compared with the prior art, the polyethylene glycol tail chain water-soluble polyphenol polymer prepared by the invention can self-assemble to form stable ultra-small nanoparticles with the potential of delivering other insoluble molecules such as drugs; due to the polyhydroxy structure, it can also be effectively coordinated with metal ions to construct a polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle T1-magnetic resonance imaging contrast agent, which has a good T1-magnetic resonance imaging effect.

Description

Technical Field

[0001] The present invention relates to the technical field of nanomaterials, in particular to a polyethylene glycol tail chain water-soluble polyphenol polymer and a preparation method and application thereof. Background Art

[0002] Polyphenols are widely found in nature, primarily derived from plants. They possess biological properties such as biocompatibility and antioxidant activity, and can be used for free radical scavenging and UV absorption. The catechol and pyrogallol moieties in many polyphenols can interact with various metal ions, molecules, or surfaces through hydrophobic bonds, hydrogen bonds, covalent bonds, electrostatic forces, π-π stacking, or metal-phenol coordination. For example, polyphenols are rich in phenolic hydroxyl groups with high affinity, making them useful as multifunctional surface coating agents. The abundant phenolic moieties in polyphenols can be utilized to chelate multivalent metal ions or coordinately self-assemble between polyphenol ligands and metal ions to form biomedical networks, which serve as contrast agents for biomedical imaging and catalysts for cellular biochemical reactions. Furthermore, due to the diverse biological functions of polyphenols, such as adhesion, antithrombotic, and anti-tumor properties, the polyphenol-metal networks formed between polyphenols and metal ions are considered promising drug delivery vehicles for cancer therapy. However, natural polyphenols have some unavoidable drawbacks. For example, polyphenols are highly unstable compounds. Their potential multi-modal stereochemistry and strong reducing properties lead to instability, which increases the complexity of their composition. In addition, although polyphenols are widely present in plants, their isolation usually requires time-consuming procedures and complex preparation conditions. Therefore, it is of great significance to construct a polyphenol with high structural stability, high reproducibility, and simple structure as a material module suitable for biological applications.

[0003] Polyethylene glycol (PEG) is a flexible, nontoxic, highly hydrophilic, simple polymer with the lowest protein or cell adsorption of any known polymer. It is physically attached to peptides, proteins, nucleic acids, liposomes, and nanoparticles to reduce renal clearance, block antibody and protein binding sites, and enhance the half-life and efficacy of therapeutics. Furthermore, the incorporation of PEG chains not only helps improve drug solubility but also creates a steric barrier between the particles and plasma proteins, reducing the formation of protein coronas and opsonization. Furthermore, the nonspecific interactions between PEG and the human body contribute to its biocompatibility. Therefore, as a standard polymer for drug delivery systems, PEG has a wide range of applications in biomedicine.

[0004] Currently, due to the rapid development of metal-polyphenol coordination polymers, many synthetic polymer nanoparticles have been explored for potential applications in biomedicine, therapy, catalysis, energy, and the environment. To achieve further in vivo delivery of bioactive moieties, including targeting ligands, drugs, prodrugs, imaging contrast agents, and tracer molecules, research on polymer-encapsulated nanoparticles continues.

[0005] Therefore, there is an urgent need for a water-soluble polyphenol polymer with a polyethylene glycol tail chain that can not only self-assemble into stable ultrasmall nanoparticles with the potential to deliver drugs and other insoluble molecules; but also effectively coordinate with metal ions to construct a magnetic resonance imaging contrast agent with good T1-magnetic resonance imaging effect. Summary of the Invention

[0006] The purpose of the present invention is to provide a polyethylene glycol tail chain water-soluble polyphenol polymer and its preparation method and application in order to overcome the defects of the above-mentioned prior art. The polyethylene glycol tail chain water-soluble polyphenol polymer prepared by the present invention can not only self-assemble to form stable ultra-small nanoparticles with the potential to deliver drugs and other insoluble molecules; but also, due to the polyhydroxy structure of the polyethylene glycol tail chain water-soluble polyphenol polymer, it can also effectively coordinate with metal ions, such as iron ions, to construct a polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle T1-magnetic resonance imaging contrast agent with good T1-magnetic resonance imaging effect.

[0007] The purpose of the present invention can be achieved by the following technical solutions:

[0008] The first object of the present invention is to provide a polyethylene glycol tail chain water-soluble polyphenol polymer, the polyethylene glycol tail chain water-soluble polyphenol polymer having the general structural formula:

[0009]

[0010] Where R is or H.

[0011] A second object of the present invention is to provide a method for preparing a polyethylene glycol tail chain water-soluble polyphenol polymer, comprising the following steps:

[0012] S11. An alkynylphenol ligand and methoxypolyethylene glycol azide are used as raw materials. Under nitrogen protection, N,N,N',N,'N"-pentamethyldiethylenetriamine (PMDETA) is added, cuprous bromide (CuBr) is used as a catalyst, and N,N-dimethylformamide (DMF) is used as a solvent. The mixture is reacted at 60-80° C. for 24-48 hours to obtain a reaction solution A.

[0013] S12, adding EDTA solution to the reaction solution A obtained in step S11, ultrafiltration and centrifugation, and freeze-drying to obtain the polyethylene glycol tail chain water-soluble polyphenol polymer;

[0014] The general structural formula of the alkynylphenol ligand is:

[0015]

[0016] The alkynylphenol ligand is alkynylphenol ligand 1 or alkynylphenol ligand 2;

[0017] In the alkynylphenol ligand 1, R is H;

[0018] In the alkynylphenol ligand 2, R is

[0019] Furthermore, the general structural formula of the methoxy polyethylene glycol azide is

[0020]

[0021] Furthermore, the concentration (volume percentage) of the EDTA solution in step S12 is 5v% to 10v%.

[0022] Furthermore, the preparation method of the alkynylphenol ligand 1 comprises the following steps:

[0023] S21, 4-propargyloxyphenol and 3,4,5-tris(tert-butyldimethylsilyloxy)benzoic acid are used as raw materials, the reaction solvent is dichloromethane, a coupling catalyst 4-pyridinium methylbenzenesulfonate (DPTS) and a hydrogen extraction base N,N'-diisopropylcarbodiimide are added at room temperature, and the reaction time is 3-5 hours to obtain a reaction solution B;

[0024] S22, filtering the reaction solution B obtained in step S21, collecting the filtrate and removing the solvent by rotary evaporation, separating and purifying by silica gel column chromatography, collecting the liquid and drying it by rotary evaporation to obtain a solid C;

[0025] S23, reacting the solid C obtained in step S22 with a mixed solution of hydrofluoric acid pyridine complex, pyridine, and tetrahydrofuran at room temperature, wherein the volume usage of the pyridine and the hydrofluoric acid pyridine complex in milliliters is 0.96 times the amount of substance of the solid C obtained in step S22 in millimolar, and the volume usage of the tetrahydrofuran in milliliters is 2 times the amount of substance of the solid C obtained in step S22 in millimolar, and the reaction time is 3-5 hours to obtain a reaction solution D;

[0026] S24. The reaction solution D obtained in step S23 is diluted with deionized water, extracted with ethyl acetate, the organic layer is washed with a saturated copper sulfate solution, dried over anhydrous magnesium sulfate, the solvent is evaporated under reduced pressure, and recrystallized using dichloromethane and methanol as developing solvents to obtain alkynylphenol ligand 1.

[0027] Furthermore, in step S21, the molar ratio of 4-propargyloxyphenol, 3,4,5-tris(tert-butyldimethylsilyloxy)benzoic acid, 4-methylbenzenesulfonate pyridine and N,N'-diisopropylcarbodiimide is 1:1.1:1.6:2.5 to 1:2:2:3.

[0028] Furthermore, the preparation method of the alkynylphenol ligand 2 comprises the following steps:

[0029] S31, using alkynylphenol ligand 1 and 3,4,5-tris(tert-butyldimethylsilyloxy)benzoic acid as raw materials, and using dichloromethane as the reaction solvent, adding a coupling catalyst 4-pyridinium methylbenzenesulfonate (DPTS) and a hydrogen extraction base N,N'-diisopropylcarbodiimide at room temperature, and reacting for 3-5 hours to obtain a reaction solution E;

[0030] S32, filtering the reaction solution E obtained in step S31, collecting the filtrate and removing the solvent by rotary evaporation, separating and purifying by silica gel column chromatography, collecting the liquid and drying it by rotary evaporation to obtain a solid F;

[0031] S33, reacting the solid F obtained in step S32 with a mixed solution consisting of a hydrofluoric acid pyridine complex, pyridine, and tetrahydrofuran at room temperature, wherein the volume amount of the pyridine and the hydrofluoric acid pyridine complex in milliliters is 0.96 times the amount of the solid F obtained in step S32 in millimolar, and the volume amount of the tetrahydrofuran in milliliters is 2 times the amount of the solid F obtained in step S32 in millimolar, and the reaction time is 3-5 hours to obtain a reaction solution G;

[0032] S34. Dilute the reaction solution G obtained in step S33 with deionized water, extract with ethyl acetate, wash the organic layer with saturated copper sulfate solution, dry over anhydrous magnesium sulfate, evaporate the solvent under reduced pressure, and recrystallize using dichloromethane and methanol as developing solvents to obtain alkynylphenol ligand 2.

[0033] Furthermore, in step S31, the molar ratio of the alkynylphenol ligand 1, 3,4,5-tris(tert-butyldimethylsilyloxy)benzoic acid, 4-methylbenzenesulfonate pyridine and N,N'-diisopropylcarbodiimide is 1:3.3:4.8:7.5 to 1:4:5:8.

[0034] Furthermore, in step S22 and step S32, the eluent used in the chromatographic separation is petroleum ether and dichloromethane in a volume ratio of 5:1.

[0035] The third object of the present invention is to provide an application of a polyethylene glycol tail chain water-soluble polyphenol polymer, wherein the polyethylene glycol tail chain water-soluble polyphenol polymer is used as a polyphenol polymer ligand to coordinate with iron ions to construct polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles, and the obtained polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solution is used as a T1-magnetic resonance imaging contrast agent and is used for T1 imaging.

[0036] Furthermore, the polyethylene glycol tail chain water-soluble polyphenol polymer is used as a polyphenol polymer ligand to coordinate with iron ions to construct polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles, and the steps of obtaining the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solution include:

[0037] S41, weighing the polyethylene glycol tail chain water-soluble polyphenol polymer and dissolving it in anhydrous ethanol to prepare a polyethylene glycol tail chain water-soluble polyphenol polymer ethanol solution;

[0038] S42, dissolving a soluble ferric salt in anhydrous ethanol until fully dissolved to obtain a ferric ion ethanol solution;

[0039] S43. Under normal temperature conditions, add the polyethylene glycol tail chain water-soluble polyphenol polymer ethanol solution to the trivalent iron ion ethanol solution, and continue stirring for 24 hours to obtain a black purple solution. After reducing the pressure to evaporate the ethanol solvent of the obtained black purple solution, add deionized water, and ultrafilter and centrifuge through a 10k ultrafiltration centrifuge tube to 20-30nm. Store at 2-8°C to obtain the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solution.

[0040] The third object of the present invention is to provide an application of a polyethylene glycol tail chain water-soluble polyphenol polymer. In step S43, the solute molar ratio of the trivalent iron ion ethanol solution to the polyethylene glycol tail chain water-soluble polyphenol polymer ethanol solution is 1:1 to 1:3.

[0041] Compared with the prior art, the present invention has the following beneficial effects:

[0042] 1) The polyethylene glycol tail chain water-soluble polyphenol polymer provided by the present invention can be used as a ligand to coordinate with iron ions to construct polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles T1-iron-based magnetic resonance imaging contrast agents, and its imaging effect is comparable to that of reported T1-iron-based magnetic resonance imaging contrast agents.

[0043] 2) The preparation method of the polyethylene glycol tail chain water-soluble polyphenol polymer provided by the present invention is simple: the reaction conditions are relatively simple, the purification process is convenient and easy to operate, and the reaction repetition efficiency is high.

[0044] 3) The polyethylene glycol tail chain water-soluble polyphenol polymer provided by the present invention has good biocompatibility because polyethylene glycol (PEG) is a flexible, non-toxic, highly hydrophilic simple polymer that has been approved by the U.S. Food and Drug Administration (FDA) for biological applications such as treatment.

[0045] 4) The polyethylene glycol tail chain water-soluble polyphenol polymer provided by the present invention has a small particle size, low toxicity, and almost no damage to tissues and organs. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 This is the hydrogen nuclear magnetic resonance spectrum of the polyethylene glycol tail chain water-soluble polyphenol polymer (R=H) in Example 1 of the present invention.

[0047] Figure 2 The polyethylene glycol tail chain water-soluble polyphenol polymer in Example 3 of the present invention H NMR spectrum.

[0048] Figure 3 This is the H NMR spectrum of methoxypolyethylene glycol azide.

[0049] Figure 4 This is a Fourier transform infrared spectrum of the polyethylene glycol tail chain water-soluble polyphenol polymer (R=H) in Example 1 of the present invention.

[0050] Figure 5 This is a transmission electron micrograph of the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles in Example 1 of the present invention.

[0051] Figure 6 This is the UV-visible spectrum of the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles in Example 1 of the present invention.

[0052] Figure 7 This is a dynamic hydration particle size distribution diagram of the polyethylene glycol tail chain water-soluble polyphenol polymer in Example 1 of the present invention.

[0053] Figure 8 This is a dynamic hydration particle size distribution diagram of the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles in Example 1 of the present invention.

[0054] Figure 9 This is a relaxation rate fitting diagram of the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles in Example 1 of the present invention under a 0.5T magnetic field and its magnetic resonance imaging contrast effect diagram.

[0055] Figure 10 This is a diagram showing the magnetic resonance imaging effect of the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles in Example 1 of the present invention under a 0.5T magnetic field. DETAILED DESCRIPTION

[0056] The present invention is described in detail below with reference to the accompanying drawings and specific embodiments. The following embodiments will help those skilled in the art further understand the present invention, but are not intended to limit the present invention in any form. It should be noted that those skilled in the art may make several variations and improvements without departing from the scope of the present invention. These all fall within the scope of protection of the present invention.

[0057] Any features such as preparation methods, materials, structures or composition ratios that are not clearly described in this technical solution shall be deemed to be common technical features disclosed in the prior art.

[0058] In the following examples, 4-propargyloxyphenol (CAS No.: 34596-27-5) and its synthesis reference: DOI No.: 10.1002 / adfm.201000729.

[0059] 3,4,5-Tris(tert-butyldimethylsilyloxy)benzoic acid, CAS number: 185462-51-5, synthesis reference: DOI: 10.1002 / chem.201200517.

[0060] Methoxypolyethylene glycol azide, CAS number: 89485-61-0.

[0061] Example 1

[0062] This embodiment provides a polyethylene glycol tail chain water-soluble polyphenol polymer, the polyethylene glycol tail chain water-soluble polyphenol polymer has the general structural formula:

[0063]

[0064] Wherein, R is H.

[0065] This embodiment further provides a method for preparing the above-mentioned polyethylene glycol tail chain water-soluble polyphenol polymer, comprising the following steps:

[0066] S11. Dissolve 4-propargyloxyphenol (148.4 mg, 1.0 mmol) and 3,4,5-tris(tert-butyldimethylsilyloxy)benzoic acid (564.2 mg, 1.1 mmol) in dichloromethane. Add coupling catalyst 4-pyridinium methylbenzenesulfonate (DPTS) (467.8 mg, 1.6 mmol) and hydrogenation base N,N'-diisopropylcarbodiimide (DIPC) (391.8 μL, 2.5 mmol) at room temperature and react for 3 hours to obtain reaction solution B.

[0067] S12. After the reaction is completed, the reaction solution B is filtered, and the filtrate is collected and evaporated to remove the solvent. The solution is separated and purified by silica gel column chromatography with an eluent of petroleum ether: dichloromethane = 5:1. The collected liquid is spin-dried and dried to obtain a solid C.

[0068] S13, then reacting the solid C obtained in step S12 with a mixed solution consisting of a hydrofluoric acid pyridine complex, pyridine, and tetrahydrofuran at room temperature, wherein the ratio of the volume usage of pyridine and the hydrofluoric acid pyridine complex in milliliters to the amount of substance of the solid C obtained in step S12 in millimoles is 0.96:1, the ratio of the volume usage of tetrahydrofuran in milliliters to the amount of substance of the solid C obtained in step S12 in millimoles is 2:1, and the reaction time is 3 hours to obtain a reaction solution D;

[0069] S14, the obtained reaction solution D is diluted with deionized water, extracted with ethyl acetate, the organic layer is washed with a saturated copper sulfate solution, dried over anhydrous magnesium sulfate, the solvent is evaporated under reduced pressure, and recrystallized with dichloromethane and methanol as developing solvents to obtain an alkynylphenol ligand (alkynylphenol ligand 1);

[0070] S15. Under nitrogen, the alkynylphenol ligand (30 mg, 0.1 mmol) and N3-PEG (200 mg, 0.1 mmol) (M w =2000) and PMDETA (20 μL, 0.1 mmol) were dissolved in 2 mL DMF; CuBr (14.4 mg, 0.1 mmol) was added; and the reaction was carried out at 60° C. for 24 hours to obtain reaction solution A. 5% EDTA solution was added to the reaction solution A to remove CuBr. + and DMF; then ultrafiltration centrifugation with deionized water to remove excess Cu + The obtained solution is freeze-dried to obtain a solid, which is the final product of the polyethylene glycol tail chain water-soluble polyphenol polymer.

[0071] This embodiment further provides an application of the above-mentioned polyethylene glycol tail-chain water-soluble polyphenol polymer, in which the above-mentioned polyethylene glycol tail-chain water-soluble polyphenol polymer is used as a polyphenol polymer ligand to coordinate with iron ions to construct polyethylene glycol tail-chain water-soluble polyphenol-iron-based nanoparticles, and the obtained polyethylene glycol tail-chain water-soluble polyphenol-iron-based nanoparticle solution is used as a T1-magnetic resonance imaging contrast agent and is used for T1 imaging.

[0072] The polyethylene glycol tail chain water-soluble polyphenol polymer is used as a polyphenol polymer ligand to coordinate with iron ions to construct polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles, and the steps of obtaining the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solution include:

[0073] S21, weighing the polyethylene glycol tail chain water-soluble polyphenol polymer and dissolving it in anhydrous ethanol to prepare a polyethylene glycol tail chain water-soluble polyphenol polymer ethanol solution;

[0074] S22, dissolving a soluble trivalent iron salt in anhydrous ethanol until fully dissolved to prepare a trivalent iron ion ethanol solution;

[0075] S23. Under normal temperature conditions, add a polyethylene glycol tail chain water-soluble polyphenol polymer ethanol solution to a trivalent iron ion ethanol solution, wherein the solute molar ratio of the trivalent iron ion ethanol solution and the polyethylene glycol tail chain water-soluble polyphenol polymer ethanol solution is 1:1, and continue stirring for 24 hours to obtain a black purple solution; after evaporating the ethanol solvent under reduced pressure, add deionized water, ultrafilter and centrifuge through a 10k ultrafiltration centrifuge tube to 20nm-30nm (about 20nm), and store at 2-8°C to obtain a polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solution.

[0076] The polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solution can be used as a T1-magnetic resonance imaging contrast agent for T1 imaging.

[0077] like Figure 1 Shown is the hydrogen nuclear magnetic resonance spectrum of the polyethylene glycol tail chain water-soluble polyphenol polymer (R=H) prepared in Example 1. Figure 1 and Figure 3 By comparison, it can be seen that H(a) of 1,4-disubstituted-1,2,3-triazole is generated after the reaction, indicating that the polyethylene glycol tail chain water-soluble polyphenol polymer (R=H) is successfully synthesized.

[0078] like Figure 4 The infrared spectrum of the polyethylene glycol tail chain water-soluble polyphenol polymer (R=H) prepared in Example 1 is shown in FIG. In the infrared spectrum, 2110 cm -1 The Vazide peak at almost disappeared, which indicated the click chemistry nature of the alkyne and azide groups, indicating that the polyethylene glycol tail chain water-soluble polyphenol polymer (R=H) was successfully synthesized.

[0079] like Figure 5 , which is a transmission electron micrograph of the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles prepared in Example 1.

[0080] like Figure 6 The UV-visible spectrum of the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles prepared in Example 1 is shown. The characteristic absorption peak of the coordination between iron ions and phenolic hydroxyl groups is at 580 cm -1 It was shown that the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles have been successfully prepared.

[0081] like Figure 7 、 8 As shown, they are the polyethylene glycol tail chain water-soluble polyphenol polymer (R=H) prepared in Example 1 ( Figure 7) and polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles ( Figure 8 ), indicating that in the absence of metal ions, the polyethylene glycol tail chain water-soluble polyphenol polymer (R = H) can form nanoparticles by self-assembly, and also indicating that the synthesized polyethylene glycol tail chain water-soluble polyphenol polymer has the potential to become a nanoscale drug delivery carrier.

[0082] The in vitro simulation experiment process of polyethylene glycol tail chain water-soluble polyphenol polymers and polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles is as follows: take polyethylene glycol tail chain polyphenol-iron-based nanoparticles with different concentrations, perform magnetic resonance relaxation measurement under magnetic fields of 0.5T and 1.0T, and test their T1-weighted MRI contrast effects under magnetic fields of 0.5T and 1.0T.

[0083] That is, polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solutions with iron ion concentrations (measured as iron ion concentration in black-purple solution) of 0, 0.05, 0.10, 0.20, 0.40, and 0.80 mM were taken, and magnetic resonance relaxation measurements were performed under a magnetic field of 0.5 T, and their T1-weighted MRI contrast effects were tested under a 0.5 T magnetic field.

[0084] like Figure 9 、 Figure 10 As shown in the figure, the relaxation rate fitting diagram of the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles prepared in Example 1 under a 0.5T magnetic field and its T1-weighted magnetic resonance imaging effect diagram are respectively shown. As can be seen from the figure, the T1 relaxation rate of the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles is 0.87mM -1 s -1 , T2 relaxation rate is 0.95mM -1 s -1 , r2 / r1=1.09, indicating that the prepared polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solution has the potential to become a T1-magnetic resonance imaging contrast agent.

[0085] Example 2

[0086] This embodiment provides a polyethylene glycol tail chain water-soluble polyphenol polymer, the polyethylene glycol tail chain water-soluble polyphenol polymer has the general structural formula:

[0087]

[0088] Wherein, R is H.

[0089] This embodiment further provides a method for preparing the above-mentioned polyethylene glycol tail chain water-soluble polyphenol polymer, comprising the following steps:

[0090] S11. Dissolve 4-propargyloxyphenol (1.588 g, 10.7 mmol) and 3,4,5-tris(tert-butyldimethylsilyloxy)benzoic acid (6.0 g, 11.8 mmol) in dichloromethane. Add coupling catalyst 4-pyridinium methylbenzenesulfonate (DPTS) (5.0 g, 17.1 mmol) and hydrogenation base N,N'-diisopropylcarbodiimide (DIPC) (4.2 ml, 26.8 mmol) at room temperature and react for 5 hours to obtain reaction solution B.

[0091] S12. After the reaction is completed, the reaction solution B is filtered, and the filtrate is collected and evaporated to remove the solvent. The solution is separated and purified by silica gel column chromatography with an eluent of petroleum ether: dichloromethane = 5:1. The collected liquid is spin-dried and dried to obtain a solid C.

[0092] S13, then reacting the solid C obtained in step S12 with a mixed solution consisting of a hydrofluoric acid pyridine complex, pyridine, and tetrahydrofuran at room temperature, wherein the ratio of the volume usage of pyridine and the hydrofluoric acid pyridine complex in milliliters to the amount of substance of the solid C obtained in step S12 in millimoles is 0.96:1, the ratio of the volume usage of tetrahydrofuran in milliliters to the amount of substance of the solid C obtained in step S12 in millimoles is 2:1, and the reaction time is 5 hours to obtain a reaction solution D;

[0093] S14, the obtained reaction solution D is diluted with deionized water, extracted with ethyl acetate, the organic layer is washed with a saturated copper sulfate solution, dried over anhydrous magnesium sulfate, the solvent is evaporated under reduced pressure, and recrystallized with dichloromethane and methanol as developing solvents to obtain an alkynylphenol ligand (alkynylphenol ligand 1);

[0094] S15. Under nitrogen, the alkynylphenol ligand (30 mg, 0.1 mmol) and N3-PEG (200 mg, 0.1 mmol) (M w =2000) and PMDETA (20 μL, 0.1 mmol) were dissolved in 2 mL DMF; CuBr (14.4 mg, 0.1 mmol) was added; the reaction was continued at 80°C for 48 hours, and 10% EDTA solution was added to remove Cu + and DMF; then ultrafiltration centrifugation with deionized water to remove excess Cu + The obtained solution is freeze-dried to obtain a solid, which is the final product of the polyethylene glycol tail chain water-soluble polyphenol polymer.

[0095] Example 3

[0096] This embodiment provides a polyethylene glycol tail chain water-soluble polyphenol polymer, the polyethylene glycol tail chain water-soluble polyphenol polymer has the general structural formula:

[0097]

[0098] Where R is

[0099] This embodiment further provides a method for preparing the above-mentioned polyethylene glycol tail chain water-soluble polyphenol polymer, comprising the following steps:

[0100] S11. Dissolve 4-propargyloxyphenol (148.4 mg, 1.0 mmol) and 3,4,5-tris(tert-butyldimethylsilyloxy)benzoic acid (564.2 mg, 1.1 mmol) in dichloromethane. Add coupling catalyst 4-pyridinium methylbenzenesulfonate (DPTS) (467.8 mg, 1.6 mmol) and hydrogenation base N,N'-diisopropylcarbodiimide (DIPC) (391.8 μL, 2.5 mmol) at room temperature and react for 3 hours to obtain reaction solution B.

[0101] S12. After the reaction is completed, the reaction solution B is filtered, and the filtrate is collected and evaporated to remove the solvent. The solution is separated and purified by silica gel column chromatography with an eluent of petroleum ether: dichloromethane = 5:1. The collected liquid is spin-dried and dried to obtain a solid C.

[0102] S13, then reacting the solid C obtained in step S12 with a mixed solution consisting of a hydrofluoric acid pyridine complex, pyridine, and tetrahydrofuran at room temperature, wherein the ratio of the volume usage of pyridine and the hydrofluoric acid pyridine complex in milliliters to the amount of substance of the solid C obtained in step S12 in millimoles is 0.96:1, the ratio of the volume usage of tetrahydrofuran in milliliters to the amount of substance of the solid C obtained in step S12 in millimoles is 2:1, and the reaction time is 3 hours to obtain a reaction solution D;

[0103] S14, diluting the obtained reaction solution D with deionized water, extracting with ethyl acetate, washing the organic layer with a saturated copper sulfate solution, drying over anhydrous magnesium sulfate, evaporating the solvent under reduced pressure, and recrystallizing with dichloromethane and methanol as developing solvents to obtain alkynylphenol ligand 1;

[0104] S15, the alkynylphenol ligand 1 (300.3 mg, 1.0 mmol) obtained in step S14 and 3,4,5-tris(tert-butyldimethylsilyloxy)benzoic acid (1.693 g, 3.3 mmol) were dissolved in dichloromethane, and a coupling catalyst 4-pyridinium methylbenzenesulfonate (DPTS) (1.413 g, 4.8 mmol) and a hydrogenation base N,N'-diisopropylcarbodiimide (DIPC) (1.2 ml, 7.5 mmol) were added at room temperature, and the mixture was reacted for 3 hours to obtain a reaction solution E;

[0105] S16. After the reaction is completed, the reaction solution E is filtered, and the filtrate is collected and evaporated to remove the solvent. The mixture is separated and purified by silica gel column chromatography with an eluent of petroleum ether: dichloromethane = 5:1. The collected liquid is spin-dried and dried to obtain a solid F.

[0106] S17, then reacting the solid F obtained in step S16 with a mixed solution consisting of a hydrofluoric acid pyridine complex, pyridine, and tetrahydrofuran at room temperature, wherein the ratio of the volume amount of pyridine and the hydrofluoric acid pyridine complex in milliliters to the amount of substance of the solid F obtained in step S16 in millimoles is 0.96:1, and the ratio of the volume amount of tetrahydrofuran in milliliters to the amount of substance of the solid F obtained in step S16 in millimoles is 2:1, and the reaction time is 3 hours to obtain a reaction solution G;

[0107] S18, the obtained reaction solution G is diluted with deionized water, extracted with ethyl acetate, the organic layer is washed with a saturated copper sulfate solution, dried over anhydrous magnesium sulfate, the solvent is evaporated under reduced pressure, and recrystallized with dichloromethane and methanol as developing solvents to obtain an alkynylphenol ligand (alkynylphenol ligand 2);

[0108] S19. Under nitrogen, the alkynylphenol ligand 2 (75.7 mg, 0.1 mmol) and N3-PEG (200 mg, 0.1 mmol) (M w =2000) and PMDETA (20 μL, 0.1 mmol) were dissolved in 2 mL DMF; CuBr (14.4 mg, 0.1 mmol) was added; and the reaction was carried out at 60° C. for 24 hours to obtain reaction solution A. 5% EDTA solution was added to the reaction solution A to remove CuBr. + and DMF; then ultrafiltration centrifugation with deionized water to remove excess Cu + The obtained solution is freeze-dried to obtain a solid, which is the final product of the polyethylene glycol tail chain water-soluble polyphenol polymer.

[0109] This embodiment further provides an application of the above-mentioned polyethylene glycol tail-chain water-soluble polyphenol polymer, in which the above-mentioned polyethylene glycol tail-chain water-soluble polyphenol polymer is used as a polyphenol polymer ligand to coordinate with iron ions to construct polyethylene glycol tail-chain water-soluble polyphenol-iron-based nanoparticles, and the obtained polyethylene glycol tail-chain water-soluble polyphenol-iron-based nanoparticle solution is used as a T1-magnetic resonance imaging contrast agent and is used for T1 imaging.

[0110] The polyethylene glycol tail chain water-soluble polyphenol polymer is used as a polyphenol polymer ligand to coordinate with iron ions to construct polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles, and the steps of obtaining the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solution include:

[0111] S21, weighing the polyethylene glycol tail chain water-soluble polyphenol polymer and dissolving it in anhydrous ethanol to prepare a polyethylene glycol tail chain water-soluble polyphenol polymer ethanol solution;

[0112] S22, dissolving a soluble trivalent iron salt in anhydrous ethanol until fully dissolved to prepare a trivalent iron ion ethanol solution;

[0113] S23. Under normal temperature conditions, add a polyethylene glycol tail chain water-soluble polyphenol polymer ethanol solution to a trivalent iron ion ethanol solution, wherein the solute molar ratio of the trivalent iron ion ethanol solution to the polyethylene glycol tail chain water-soluble polyphenol polymer ethanol solution is 1:3, and continue stirring for 24 hours to obtain a black purple solution; after evaporating the ethanol solvent under reduced pressure, add deionized water, ultrafilter and centrifuge through a 10k ultrafiltration centrifuge tube to 20-30nm (about 20nm), and store at 2-8°C to obtain a polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solution.

[0114] The polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solution can be used as a T1-magnetic resonance imaging contrast agent (polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle T1-magnetic resonance imaging contrast agent) for T1 imaging.

[0115] like Figure 2 Shown is the hydrogen nuclear magnetic resonance spectrum of the polyethylene glycol tail chain water-soluble polyphenol polymer (R=H) prepared in Example 3. Figure 2 and Figure 3 By comparison, we can see the H(a) of 1,4-disubstituted-1,2,3-triazole generated after the reaction, indicating that the polyethylene glycol tail chain water-soluble polyphenol polymer Successfully synthesized.

[0116] Comparative Example 1

[0117] An in vitro simulation experiment was conducted using GA-Fe@BSA as a contrast agent (same as in Example 1). The results are shown in the following table in comparison with the contrast agent described in Example 1:

[0118] <![CDATA[T1 relaxation rate]]> <![CDATA[T2 relaxation rate]]> Example 1 <![CDATA[0.87mM -1 s -1 ]]> <![CDATA[0.95mM -1 s -1 ]]> Comparative Example 1 <![CDATA[0.79mM -1 s -1 ]]> <![CDATA[0.87mM -1 s -1 ]]>

[0119] Comparing Example 1 and Comparative Example 1, the imaging effect of the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solution of Example 1 as a T1-magnetic resonance imaging contrast agent is comparable to the imaging effect of the reported T1-magnetic resonance imaging contrast agent GA-Fe@BSA.

[0120] The above description of the embodiments is intended to facilitate understanding and use of the invention by those skilled in the art. It will be apparent that those skilled in the art can readily make various modifications to these embodiments and apply the general principles described herein to other embodiments without requiring inventive effort. Therefore, the present invention is not limited to the above-described embodiments. Improvements and modifications made by those skilled in the art based on the disclosure of the present invention, without departing from the scope of the present invention, should be within the scope of protection of the present invention.

Claims

1. A polyethylene glycol tail chain water-soluble polyphenol polymer, characterized in that: The general structural formula of the polyethylene glycol tail chain water-soluble polyphenol polymer is: Among them, R is or H.

2. A method for preparing a polyethylene glycol tail chain water-soluble polyphenol polymer as claimed in claim 1, characterized in that: The following steps are involved: S11, using alkynylphenol ligand and methoxy polyethylene glycol azide as raw materials, adding N,N,N',N,'N"-pentamethyldiethylenetriamine under nitrogen protection, using cuprous bromide as a catalyst, and N,N-dimethylformamide as a solvent, reacting at 60-80° C. for 24-48 hours to obtain a reaction solution A; S12, adding EDTA solution to the reaction solution A obtained in step S11, ultrafiltration and centrifugation, and freeze-drying to obtain the polyethylene glycol tail chain water-soluble polyphenol polymer; The general structural formula of the alkynylphenol ligand is: The alkynylphenol ligand is alkynylphenol ligand 1 or alkynylphenol ligand 2; In the alkynylphenol ligand 1, R is H; In the alkynylphenol ligand 2, R is 3. The method for preparing a polyethylene glycol tail chain water-soluble polyphenol polymer according to claim 2, characterized in that: The concentration of the EDTA solution in step S12 is 5v% to 10v%.

4. The method for preparing a polyethylene glycol tail chain water-soluble polyphenol polymer according to claim 2, characterized in that: The preparation method of the alkynylphenol ligand 1 comprises the following steps: S21, using 4-propargyloxyphenol and 3,4,5-tri(tert-butyldimethylsilyloxy)benzoic acid as raw materials, using dichloromethane as the reaction solvent, adding coupling catalyst 4-pyridinium toluenesulfonate (DPTS) and hydrogen extraction base N,N'-diisopropylcarbodiimide at room temperature, reacting for 3-5 hours, and obtaining reaction solution B; S22, filtering the reaction solution B obtained in step S21, collecting the filtrate and removing the solvent by rotary evaporation, separating and purifying by silica gel column chromatography, collecting the liquid and rotary drying, and drying to obtain a solid C; S23, reacting the solid C obtained in step S22 with a mixed solution consisting of a hydrofluoric acid pyridine complex, pyridine, and tetrahydrofuran at room temperature, wherein the volume usage of the pyridine and the hydrofluoric acid pyridine complex in milliliters is 0.96 times the amount of the substance in millimole of the solid C obtained in step S22, and the volume usage of the tetrahydrofuran in milliliters is 2 times the amount of the substance in millimole of the solid C obtained in step S22, and the reaction time is 3-5 hours to obtain a reaction solution D; S24, diluting the reaction solution D obtained in step S23 with deionized water, extracting with ethyl acetate, washing the organic layer with a saturated copper sulfate solution, drying with anhydrous magnesium sulfate, evaporating the solvent under reduced pressure, and recrystallizing with dichloromethane and methanol as developing agents to obtain alkynylphenol ligand 1.

5. The method for preparing a polyethylene glycol tail chain water-soluble polyphenol polymer according to claim 4, characterized in that: In step S21, the molar ratio of 4-propargyloxyphenol, 3,4,5-tri(tert-butyldimethylsilyloxy)benzoic acid, 4-methylbenzenesulfonate pyridine and N,N'-diisopropylcarbodiimide is 1:1.1:1.6:2.5 to 1:2:2:

3.

6. The method for preparing a polyethylene glycol tail chain water-soluble polyphenol polymer according to claim 4, characterized in that: The preparation method of the alkynylphenol ligand 2 comprises the following steps: S31, using alkynylphenol ligand 1 and 3,4,5-tri(tert-butyldimethylsilyloxy)benzoic acid as raw materials, using dichloromethane as the reaction solvent, adding coupling catalyst 4-pyridinium methylbenzenesulfonate (DPTS) and hydrogen extraction base N,N'-diisopropylcarbodiimide at room temperature, reacting for 3-5 hours, and obtaining reaction solution E; S32, filtering the reaction solution E obtained in step S31, collecting the filtrate and removing the solvent by rotary evaporation, separating and purifying by silica gel column chromatography, collecting the liquid and rotary drying, and drying to obtain a solid F; S33, reacting the solid F obtained in step S32 with a mixed solution of hydrofluoric acid pyridine complex, pyridine and tetrahydrofuran at room temperature, wherein the volume amount of the pyridine and the hydrofluoric acid pyridine complex in milliliters is 0.96 times the amount of the solid F obtained in step S32 in millimoles, and the volume amount of the tetrahydrofuran in milliliters is 2 times the amount of the solid F obtained in step S32 in millimoles, and the reaction time is 3-5 hours to obtain a reaction solution G; S34, diluting the reaction solution G obtained in step S33 with deionized water, extracting with ethyl acetate, washing the organic layer with saturated copper sulfate solution, drying with anhydrous magnesium sulfate, evaporating the solvent under reduced pressure, and recrystallizing with dichloromethane and methanol as developing agents to obtain alkynylphenol ligand 2.

7. The method for preparing a polyethylene glycol tail chain water-soluble polyphenol polymer according to claim 6, characterized in that: In step S31, the molar ratio of the alkynylphenol ligand 1, 3,4,5-tri(tert-butyldimethylsilyloxy)benzoic acid, 4-methylbenzenesulfonate pyridine and N,N'-diisopropylcarbodiimide is 1:3.3:4.8:7.5 to 1:4:5:

8.

8. An application of the polyethylene glycol tail chain water-soluble polyphenol polymer as claimed in claim 1 or the polyethylene glycol tail chain water-soluble polyphenol polymer prepared by the preparation method of the polyethylene glycol tail chain water-soluble polyphenol polymer as described in any one of claims 2 to 7, using the polyethylene glycol tail chain water-soluble polyphenol polymer as a polyphenol polymer ligand to coordinate with iron ions to construct polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles and using the obtained polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solution as a T1-magnetic resonance imaging contrast agent and for T1 imaging.

9. The use of a polyethylene glycol tail chain water-soluble polyphenol polymer according to claim 8, characterized in that: The steps of using the polyethylene glycol tail chain water-soluble polyphenol polymer as a polyphenol polymer ligand to coordinate with iron ions to construct polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles to obtain the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solution include: S41, weighing the polyethylene glycol tail chain water-soluble polyphenol polymer and dissolving it in anhydrous ethanol to prepare a polyethylene glycol tail chain water-soluble polyphenol polymer ethanol solution; S42, dissolving a soluble trivalent iron salt in anhydrous ethanol, and fully dissolving it to obtain a trivalent iron ion ethanol solution; S43. Under normal temperature conditions, add a polyethylene glycol tail chain water-soluble polyphenol polymer ethanol solution to a trivalent iron ion ethanol solution, and continue stirring for 24 hours to obtain a dark purple solution. After reducing the pressure to evaporate the ethanol solvent of the obtained dark purple solution, add deionized water, and ultrafilter and centrifuge through a 10k ultrafiltration centrifuge tube until the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticles are 20-30nm, and store at 2-8°C to obtain the polyethylene glycol tail chain water-soluble polyphenol-iron-based nanoparticle solution.

10. The use of a polyethylene glycol tail chain water-soluble polyphenol polymer according to claim 9, characterized in that: In step S43, the solute molar ratio of the ferric iron ion ethanol solution to the polyethylene glycol tail chain water-soluble polyphenol polymer ethanol solution is 1:1 to 1:3.

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