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CN-122005767-A - Microenvironment programming nano-enzyme and diabetes wound infection medicine thereof

CN122005767ACN 122005767 ACN122005767 ACN 122005767ACN-122005767-A

Abstract

The invention provides microenvironment programming nano-enzyme and a diabetes wound infection medicament, and belongs to the technical field of biopharmaceuticals. The nano-enzyme takes copper-chlorogenic acid self-assembled nano-fiber (CuCA) as a core carrier, L-arginine and glucose oxidase are loaded on the surface of the nano-enzyme, and dextran is coated on the outer layer of the nano-enzyme. The system has the response characteristics of intelligent microenvironment, and is capable of generating hydroxyl free radicals through cascade catalysis in an acidic infection environment, realizing efficient sterilization and toxicity down regulation by combining with quorum sensing inhibition effect, and utilizing catalase-like activity to remove residual oxygen and supply oxygen in a neutral repair stage to cooperatively promote angiogenesis. The invention solves the problem of difficult healing of diabetes wounds caused by drug-resistant bacteria infection, realizes the programmed treatment of antibacterial, anti-inflammatory and repair promotion, has high healing rate and good biological safety, and has wide application prospect in the field of biological medicine.

Inventors

  • XIANG YI
  • CAI WEIYI
  • LIU YANAN
  • LI JIAHUI

Assignees

  • 深圳市龙华区妇幼保健院(深圳市龙华区婴幼儿照护服务指导中心、深圳市龙华区健康教育所)

Dates

Publication Date
20260512
Application Date
20260414

Claims (10)

  1. 1. A micro-environment programming nano-enzyme, characterized by comprising the following steps: The core carrier is nano fiber CuCA formed by self-assembly of copper ions and chlorogenic acid; l-arginine and glucose oxidase supported on the surface of the core carrier, and And the dextran layer is coated on the outermost layer.
  2. 2. The microenvironment programmed nanoenzyme of claim 1, wherein: the molar ratio of copper ions to chlorogenic acid in CuCA is 1:1 to 2:1; The mass ratio of the L-arginine to CuCA is 1:1 to 1:2; The mass ratio of the glucose oxidase to the CuCA loaded with L-arginine is 1:2 to 1:3; The mass ratio of the glucan layer to CuCA loaded with L-arginine and glucose oxidase is 1:1 to 1:2.
  3. 3. The microenvironment-programmed nanoenzyme of claim 1, wherein said nanoenzyme has microenvironment-responsive catalytic activity: In the environment of pH 5.0-6.5, the enzyme shows peroxidase activity, and H 2 O 2 generated by catalyzing glucose oxidation is converted into hydroxyl free radical; At pH 7.0-7.4, exhibits catalase activity, and catalyzes the decomposition of H 2 O 2 to produce O 2 .
  4. 4. The microenvironment programmed nanoenzyme of claim 1, wherein said nanoenzyme is capable of down-regulating expression of an agr quorum sensing system related gene of a drug resistant staphylococcus aureus.
  5. 5. A method for preparing the microenvironment programmed nanoenzyme according to any one of claims 1 to 4, comprising the following steps: s1, dissolving a copper source and a dispersing agent in water, regulating pH to be alkaline, and adding chlorogenic acid solution to perform self-assembly reaction to obtain a core carrier CuCA; S2, mixing CuCA with L-arginine in a buffer solution, and obtaining an intermediate Cu-L through electrostatic adsorption; s3, carrying out amidation reaction on an intermediate Cu-L and glucose oxidase under a condensing agent system to obtain a cascade catalytic system Cu-L-G; S4, mixing the Cu-L-G with the glucan solution, and coating the glucan solution to obtain the microenvironment programming nano-enzyme.
  6. 6. The method according to claim 5, wherein the alkaline pH in step S1 is 12-14, and the self-assembly reaction time is 10-14 hours.
  7. 7. The process according to claim 5, wherein the amidation reaction in step S3 is carried out at a temperature of 35 to 40℃for 20 to 28 hours.
  8. 8. The method of manufacturing according to claim 5, wherein: In the step S1, the copper source is copper chloride, and the dispersing agent is polyvinylpyrrolidone; In step S3, the condensing agent is EDC and NHS.
  9. 9. The use of a microenvironment programmed nanoenzyme according to any one of claims 1-4 for the preparation of an antibacterial drug for inhibiting the formation of a bacterial biofilm or for disrupting a mature biofilm.
  10. 10. Use of the microenvironment-programmed nanoenzyme of any one of claims 1-4 in the preparation of a medicament for promoting diabetic wound healing.

Description

Microenvironment programming nano-enzyme and diabetes wound infection medicine thereof Technical Field The invention belongs to the technical field of biological pharmacy, and particularly relates to microenvironment programming nano-enzyme and a diabetes wound infection medicament. Background Chronic wound infection caused by diabetic complications has become a major clinical problem to be overcome in the world public health field. Epidemiological data indicate that about 15% of diabetics worldwide experience skin ulcers or wound infections during their course, with up to 33% of patients facing amputation risks due to prolonged wound healing, severely threatening the quality of life and life safety of the patient. The difficult healing of diabetic wounds is caused by extremely complex pathological microenvironment, namely, the peripheral vascular lesions induced by the long-term high-sugar state cause local microcirculation disturbance, so that the wounds are in serious hypoxia and nutrient deficiency states for a long time. Meanwhile, excessive accumulated Reactive Oxygen Species (ROS) in the microenvironment induces severe oxidative stress, resulting in oxidative damage of proteins, lipids and DNA. More importantly, sustained oxidative stress drives macrophage polarization to pro-inflammatory M1 type, induces massive secretion of inflammatory factors, causes long-term retention of the wound in chronic inflammatory phase, and seriously hinders physiological transformation from inflammatory phase to tissue repair phase. In the high-sugar, low-oxygen biochemical environment of diabetic wounds, drug-resistant bacterial infection and the consequent biofilm (Biofilm) are the core mechanisms leading to failure of conventional therapies. Pathogenic bacteria represented by methicillin-resistant staphylococcus aureus (MRSA) are precisely regulated in pathogenicity by a Quorum Sensing (QS) system. For example, the agr system of MRSA dynamically regulates expression of virulence factors such as hemolysin, exoproteinase, etc. by sensing signal molecule concentration, and induces complex extracellular polymeric matrix production, thereby constructing a biofilm structure. The biofilm acts as a physical barrier, increasing the tolerance of internal bacteria to traditional antibiotics by hundreds of times, resulting in a resistance to drug penetration. In addition, MRSA can synthesize staphyloxanthin as a biochemical defense barrier, and can effectively remove oxidative killer factors released by host immune cells. Thus, single sterilization strategies tend to be difficult to penetrate the biofilm barrier and do not effectively interfere with bacterial virulence metabolism, which has become a major technical bottleneck in current clinical anti-infective regimens. The nano-enzyme catalytic therapy which has been in progress in recent years provides a new research paradigm for breaking the dilemma. The nano-enzyme is used as a nano-material with enzyme-like catalytic activity, and can simulate natural enzymes (such as POD, CAT, SOD and the like) to trigger cascade chemical reactions in situ at lesion sites. On the one hand, the nano-enzyme with Peroxidase (POD) activity can convert H 2O2 in the bacterial microenvironment into highly toxic hydroxyl free radicals (OH) through Fenton-like reaction, so as to realize physical sterilization independent of antibiotics. However, the endogenous H 2O2 level of the wound surface fluctuates severely and often has insufficient concentration, which limits the continuous sterilization efficacy of the single-function nano-enzyme. On the other hand, excessive ROS remained after sterilization can further aggravate tissue oxidative damage if not timely cleared. Nanoezymes with Catalase (CAT) activity, while capable of scavenging ROS and producing O 2 to alleviate hypoxia, often have a logical conflict in their function with the antibacterial pattern in the same space-time. Although various multifunctional nano platforms are proposed at present, how to construct an intelligent system capable of sensing micro-environment changes and realizing accurate function switching aiming at pathological features of different stages of diabetes infected wounds is still the direction of attack of the current research. In particular, the ideal therapeutic regimen should have the "programmed" characteristics of being able to efficiently generate reactive oxygen species and interfere with bacterial quorum sensing at the early stages of infection (acidic, high-sugar microenvironment), inhibiting virulence factor expression and biofilm formation, and rapidly switching to antioxidant mode during the repair phase after infection control (neutral microenvironment), promoting angiogenesis and granulation tissue growth by scavenging excess ROS, relieving hypoxia. The development of the nanometer therapeutic system integrating high-efficiency antibacterial and antitoxic force regulation and space-time programming rep