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CN-121971371-A - Puerarin-polypeptide co-assembled nano hydrogel and preparation method and application thereof

CN121971371ACN 121971371 ACN121971371 ACN 121971371ACN-121971371-A

Abstract

The invention discloses puerarin-polypeptide co-assembled nano hydrogel, and belongs to the technical field of biological medicines. Aiming at the problems of poor water solubility and stability of puerarin and difficult healing of diabetic wounds, the hydrogel comprises self-assembled polypeptide and puerarin in a mass ratio of 1:0.5-4, wherein the molecular formula of the self-assembled polypeptide is R-G D F D F D Y, D , the amino acid is in a D configuration, and R is a nonsteroidal anti-inflammatory drug. The self-assembled polypeptide and puerarin are assembled together to form the nano hydrogel, so that the solubility and stability of the puerarin are improved, and the self-assembled polypeptide and puerarin can be used for preparing medicines for healing wounds of diabetes.

Inventors

  • LI XINXIN
  • HUANG GUANGRUI
  • SHAO BEIBEI

Assignees

  • 北京中医药大学

Dates

Publication Date
20260505
Application Date
20251230

Claims (8)

  1. 1. The puerarin-polypeptide co-assembled nano hydrogel is characterized by comprising self-assembled polypeptide and puerarin in a mass ratio of 1:0.5-4, wherein the molecular formula of the self-assembled polypeptide is R-G D F D F D Y, D , the amino acid is in a D configuration, and R is a nonsteroidal anti-inflammatory drug.
  2. 2. The method for preparing puerarin-polypeptide co-assembled nano-hydrogel according to claim 1, comprising the following steps: adding the self-assembled polypeptide and puerarin into phosphate buffer solution, mixing, heating, and adjusting the pH of the mixed solution to 7-8 by using sodium carbonate solution until the mixed solution is completely dissolved, and standing and cooling at room temperature to form the puerarin-polypeptide co-assembled nano hydrogel, wherein the mass ratio of the self-assembled polypeptide to the puerarin is 1-4:0.75-7.5.
  3. 3. The method for preparing puerarin-polypeptide co-assembled nano-hydrogel according to claim 2, wherein the self-assembled polypeptide comprises a non-steroidal anti-inflammatory drug and a short peptide G D F D F D Y.
  4. 4. The method for preparing puerarin-polypeptide co-assembled nano-hydrogel according to claim 3, wherein the method for preparing the self-assembled polypeptide comprises the following steps: s1, dissolving N- (9-fluorenylmethoxycarbonyl) -O-tertiary butyl-D-tyrosine by using methylene dichloride, adding diisopropylethylamine, and mixing to obtain a reaction liquid I; s2, fully swelling the 2-Cl-Trt resin by adopting anhydrous dichloromethane, removing the dichloromethane, adding the reaction liquid I into a solid-phase synthesizer, reacting on a shaking table at room temperature, and obtaining a mixture 1 after the reaction is finished; S3, removing liquid in the mixture 1, washing for multiple times by adopting anhydrous dichloromethane, adding a sealing liquid, reacting on a shaking table at room temperature, and obtaining a mixture 2 after the reaction is finished, wherein the sealing liquid comprises dichloromethane, N diisopropylethylamine, methanol=17:1:2; S4, removing liquid in the mixture 2, sequentially adopting anhydrous dichloromethane and N, N-dimethylformamide to wash for multiple times, then adding a piperidine solution, reacting at room temperature, removing the liquid after the reaction is finished, and adopting N, N-dimethylformamide to wash for multiple times, wherein the solvent of the piperidine solution is N, N-dimethylformamide; S5, adding O-benzotriazole-tetramethyl urea hexafluorophosphate, diisopropylethylamine and N, N-dimethylformamide into N- (9-fluorenylmethoxycarbonyl) -D-phenylalanine, dissolving to obtain a reaction solution II, adding the reaction solution II into the solid phase washed in the step four, and reacting in a solid phase synthesizer until the reaction is finished; S6, repeating the step S4 and the step S5, sequentially adding N- (9-fluorenylmethoxycarbonyl) -L-phenylalanine and N- (9-fluorenylmethoxycarbonyl) -glycine, washing for a plurality of times by adopting N, N-dimethylformamide after the last amino acid is grafted, adding a piperidine solution for room temperature reaction, removing liquid after the reaction is finished, and washing for a plurality of times by adopting N, N-dimethylformamide to obtain a short peptide G D F D F D Y; s7, adding a peptide coupling reagent HBTU, diisopropylethylamine and N, N-dimethylformamide into a non-steroidal anti-inflammatory drug, dissolving to obtain a reaction solution III, reacting the reaction solution III with a short peptide G D F D F D Y in a solid-phase synthesizer, removing the liquid after the reaction is finished, and then sequentially adopting the N, N-dimethylformamide and anhydrous dichloromethane to wash for a plurality of times to obtain an intermediate product; s8, adding a cutting fluid into the intermediate product of the solid phase synthesizer to cut R-G D F D F D Y from 2-Cl-Trt resin, concentrating and drying to obtain a crude product, and purifying the crude product to obtain the self-assembled polypeptide.
  5. 5. The method for preparing puerarin-polypeptide co-assembled nano-hydrogel according to claim 4, wherein the volume fraction of the piperidine solution in the step S4 and the step S6 is 20-40%, and the solvent of the piperidine solution is N, N-dimethylformamide.
  6. 6. The method for preparing puerarin-polypeptide co-assembled nano-hydrogel according to claim 4, wherein the cutting fluid comprises trifluoroacetic acid and triisopropylsilane in a volume ratio of water=95:2.5:2.5.
  7. 7. The use of puerarin-polypeptide co-assembled nano-hydrogel according to claim 1 in the preparation of a medicament for healing wounds of diabetes.
  8. 8. The use according to claim 7, wherein the puerarin-polypeptide co-assembled nano-hydrogel promotes wound healing by modulating macrophage polarization, reducing AGEs accumulation at the wound site, restoring blood circulation and remodeling.

Description

Puerarin-polypeptide co-assembled nano hydrogel and preparation method and application thereof Technical Field The invention relates to the technical field of biological medicine. More specifically, the invention relates to puerarin-polypeptide co-assembled nano hydrogel and a preparation method and application thereof. Background The treatment of wounds, especially chronic wounds that are increasingly severe due to increased incidence of obesity and diabetes, resulting from trauma, burns, surgery and chronic diseases presents a great challenge, such wound repair involves multi-stage complex biological processes such as inflammatory regulation, cell proliferation, angiogenesis, etc., traditional invasive treatments such as surgical debridement and skin grafting, not only involve the risks of bleeding, infection, etc., but also require long-term bedridden recovery, and single treatment costs can reach tens of thousands of dollars, giving a dual burden to patients and medical systems. The non-invasive gel dressing has a three-dimensional network structure similar to soft tissues, injectable in-situ gel forming property and drug targeting delivery function, so that the non-invasive gel dressing becomes a research hot spot for wound treatment. The current hydrogel dressing based on biological materials is of various types, including peptide-based, chitosan, collagen and other systems. Wherein, the supermolecular hydrogel based on self-assembled polypeptide has a fiber network structure highly similar to fibrin in extracellular matrix (ECM), can provide bionic microenvironment for cell adhesion and migration, and has unique advantages in promoting tissue repair. For example, GFFY-motif short peptide self-assembled nanomaterials have been demonstrated to promote antigen presentation and lymph node enrichment by activating humoral and cellular immunity, and their successful application in the fields of tumor immunotherapy and vaccine adjuvants provides a new concept for wound healing dressing design. Puerarin is used as natural flavonoid extracted from radix Puerariae, and has multiple biological activities such as antioxidant, antiinflammatory, and glycolipid metabolism regulating effects. Research shows that the compound can reduce oxidative stress injury by inhibiting NADPH oxidase activity, down regulate the expression of proinflammatory factors such as TNF-alpha, IL-6 and the like, reduce accumulation of advanced glycosylation end products (AGEs), improve insulin resistance, and have definite curative effects in preventing and treating diabetes and complications thereof. However, puerarin lacks ideal carrier in wound repair application, free puerarin is easy to degrade or metabolize rapidly to cause insufficient local drug concentration, while traditional dosage forms such as vaseline ointment have defects of uncontrollable drug release, adhesive property of dressing, water-retaining property and the like, which severely limit the clinical transformation in wound healing. The existing hydrogel dressing based on a single component can partially solve the carrier problem, but still has the limitation of single function, namely, the pure polypeptide hydrogel lacks the antioxidant and blood sugar regulating capability and is difficult to cope with the oxidative stress microenvironment of the diabetes wound, and the puerarin cannot fully exert the multi-target treatment advantage due to low delivery efficiency when being singly applied. Therefore, a composite hydrogel system with self-assembly performance and drug co-loading function is constructed, and high-efficiency delivery and function synergy of puerarin are realized, so that the composite hydrogel system becomes a key technical difficulty for breaking through the bottleneck of diabetes wound healing treatment. Disclosure of Invention It is an object of the present invention to solve at least the above problems and to provide at least the advantages to be described later. To achieve these objects and other advantages and in accordance with the purpose of the invention, there is provided a puerarin-polypeptide co-assembled nano-hydrogel comprising self-assembled polypeptide and puerarin in a mass ratio of 1:0.5-4, wherein the molecular formula of the self-assembled polypeptide is R-G DFDFDY,D, which represents that amino acid is in D configuration, and R is a non-steroidal anti-inflammatory drug. The preparation method of the puerarin-polypeptide co-assembled nano hydrogel comprises the following steps: adding the self-assembled polypeptide and puerarin into phosphate buffer solution, mixing, heating, and adjusting the pH of the mixed solution to 7-8 by using sodium carbonate solution until the mixed solution is completely dissolved, and standing and cooling at room temperature to form the puerarin-polypeptide co-assembled nano hydrogel, wherein the mass ratio of the self-assembled polypeptide to the puerarin is 1-4:0.75-7.5. Preferably, the self-assembling p