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CN-122005490-A - Drug delivery system of targeted neutrophil extracellular trap net and preparation method and application thereof

CN122005490ACN 122005490 ACN122005490 ACN 122005490ACN-122005490-A

Abstract

The invention discloses a drug delivery system of a targeted neutrophil extracellular trap net, a preparation method and application thereof, wherein the drug delivery system takes superparamagnetic iron oxide nanoparticles as cores and mesoporous polydopamine as shells to form a Fe 3 O 4 @mPDA core-shell structure carrier, DOX and DNase I are loaded by using a mesoporous structure of the carrier, and tumor cell membrane fragments of CCDC25 are coated to form nanoparticles with a targeted recognition function. The drug delivery system can realize the efficient enrichment of the NETs region by specifically recognizing and combining DNA components in the NETs through CCDC25, generates a photo-thermal effect under the irradiation of near infrared laser, controllably releases drug molecules DOX and DNase I, and can degrade the DNA skeleton of the NETs to destroy the metastasis-promoting microenvironment so as to inhibit tumor cell metastasis, and the DOX plays a role in chemotherapy and generates a synergistic anti-tumor effect with photo-thermal treatment.

Inventors

  • XIE LIQIN
  • Yuan Fenghou
  • CHEN HONGLI
  • NAN WENBIN
  • ZHANG TAO
  • HAO YONGWEI

Assignees

  • 河南医药大学

Dates

Publication Date
20260512
Application Date
20260126

Claims (7)

  1. 1. A drug delivery system of a targeted neutrophil extracellular trap net is characterized in that superparamagnetic ferroferric oxide (Fe 3 O 4 ) nanoparticles are adopted as cores in the drug delivery system, mesoporous polydopamine (mPDA) shells are constructed on the surfaces of the drug delivery system, a nano carrier Fe 3 O 4 @mPDA with a core-shell structure is formed, the nano carrier Fe 3 O 4 @mPDA utilizes mesoporous structure to efficiently co-load a drug molecule chemotherapeutic drug Doxorubicin (DOX) and a deoxyribonuclease I (DNase I) for degrading the neutrophil extracellular trap net, drug-loaded nanoparticles are obtained, and targeting molecule coiled helical domain protein 25 (CCDC 25) capable of specifically recognizing and combining the neutrophil extracellular trap net is modified on the surfaces of the drug-loaded nanoparticles, so that the drug delivery system of the targeted neutrophil extracellular trap net is constructed, and the drug delivery system targets the neutrophil extracellular trap net in a tumor micro-environment, promotes release of drug molecules under the action of photo-thermal therapy and realizes synergistic effect of chemotherapy and photo-degradation of the tumor, and simultaneously degrades the tumor-loaded tumor micro-tumor pre-transferred micro-tumor environment before the tumor cell extracellular trap net is transferred.
  2. 2. The drug delivery system for targeting the neutrophil extracellular trap according to claim 1, wherein the surface of the drug delivery system is modified with CCDC25, the CCDC25 can specifically identify and combine deoxyribonucleic acid (DNA) components in the neutrophil extracellular trap, so that the drug delivery system can actively target to a tumor microenvironment area enriched by the neutrophil extracellular trap, the ferroferric oxide enables the drug delivery system to have magnetic response capability, can stay at a focus part under the action of an external magnetic field, improves local drug concentration, the mesoporous polydopamine shell has near infrared thermal conversion performance, can generate photo-thermal effect under the irradiation of near infrared light and further forms synergistic anti-tumor effect with the chemotherapeutic action of DOX, DNase I is protected in a nano carrier Fe 3 O 4 @mPDA, the in vivo stability and half-life are improved, the degradation efficiency of the DNA skeleton of the neutrophil extracellular trap is enhanced, the microenvironment before tumor transfer is destroyed, and the tumor growth of the drug delivery system can be inhibited and prevented from primary tumor growth by the multiple synergistic effect of chemotherapy and photo-thermal treatment and targeted neutrophil extracellular trap degradation.
  3. 3. A method of preparing a drug delivery system for targeting neutrophil extracellular traps according to claim 1 or 2, characterized by the specific preparation steps of: step S1, preparation of superparamagnetic ferroferric oxide nanoparticles Dispersing ferric chloride hexahydrate, sodium acetate and sodium citrate dihydrate in ethylene glycol, stirring and mixing uniformly, reacting at 180-220 ℃ in a reaction kettle, washing the product with ethanol and distilled water respectively after the reaction is finished, and performing magnetic separation to obtain superparamagnetic ferroferric oxide nanoparticles; Step S2, preparation of nano carrier Fe 3 O 4 @ mPDA Adding pluronic F127 and 1,3, 5-trimethylbenzene into a mixed solution of deionized water and ethanol, fully stirring until emulsification, sequentially adding tris (hydroxymethyl) aminomethane, dopamine (PODA) and superparamagnetic ferroferric oxide nanoparticles obtained in the step S1, reacting at room temperature, magnetically separating a product after the reaction is finished, and washing a precipitate with a mixed solution of absolute ethanol and acetone to obtain a nano carrier Fe 3 O 4 @mPDA with a core-shell structure; Step S3, preparation of drug-loaded nano-particles Fe 3 O 4 @ mPDA-DOX-DNase I Dispersing the nano carrier Fe 3 O 4 @mPDA obtained in the step S2 in deionized water, adding an aqueous solution of DOX, stirring and reacting at 2-6 ℃ in a dark place, and centrifuging the product after the reaction is finished to obtain drug-loaded nano particles Fe 3 O 4 @mPDA-DOX; Step S4, modification of membrane protein CCDC25 on drug-loaded nanoparticles Incubating virus liquid containing CCDC25 with 4T1 cells, transfecting for 24 hours, screening and obtaining transfected positive cells to obtain 4T1 cells over-expressed by the CCDC25, marking the 4T1 cells as C-4T1 cells, amplifying the C-4T1 cells, adding an extraction reagent A containing a Cocktail protease inhibitor into the 4T1 cells, repeatedly freezing and thawing after incubation to break the cells, centrifugally collecting cell membrane fragments containing the CCDC25, adding a membrane protein extraction reagent B, fully extracting the protein CCDC25, centrifugally collecting supernatant of the membrane protein CCDC25 at a high speed, placing the membrane protein CCDC25 and drug-loaded nano particles Fe 3 O 4 @mPDA-DOX-DNase I into a liposome extruder, and magnetically separating the product to obtain a drug-loaded nano particle modified by the CCDC25, namely a drug delivery system of a targeted neutrophil extracellular trapping net, marking the drug-loaded nano particles as CCDC25-Fe 3 O 4 @mPDA-DOX-DNase I.
  4. 4. The preparation method of the drug delivery system of the targeted neutrophil extracellular trap according to claim 3, which is characterized in that in the step S4, the membrane protein CCDC25 is modified by drug-carrying nanoparticles, wherein the specific process comprises the steps of respectively taking the membrane protein CCDC25 and the drug-carrying nanoparticles Fe 3 O 4 @mPDA-DOX-DNase I according to a mass ratio of 1:1000, respectively preparing a protein solution and a drug-carrying nanoparticle aqueous solution, respectively placing the protein solution and the drug-carrying nanoparticle aqueous solution on two sides of a liposome extruder, extruding for 8-12 times, and then carrying out magnetic separation to obtain the drug delivery system CCDC25-Fe 3 O 4 @mPDA-DOX-DNase I.
  5. 5. Use of a drug delivery system targeting an extracellular trap of neutrophils according to claim 1 or 2 for the preparation of a medicament for the treatment and/or prevention of tumors and tumor metastases.
  6. 6. The application of the medicine according to claim 5, wherein the medicine is characterized in that the medicine acts by retaining ferroferric oxide nuclear hysteresis at focus positions to improve local medicine concentration, guiding a medicine delivery system by targeting molecule CCDC25 to be specifically enriched in a neutrophil extracellular trap net area in a tumor microenvironment, degrading a DNA skeleton of the neutrophil extracellular trap net by DNase I to destroy a metastasis promotion microenvironment, killing tumor cells by chemotherapeutic medicine DOX, generating a photo-thermal effect by mesoporous polydopamine shell under near infrared light irradiation, and killing the tumor cells.
  7. 7. Use of a drug delivery system of a targeted neutrophil extracellular trap according to claim 1 or 2 for the preparation of a drug for the synergistic treatment of tumors by chemotherapy-photothermal therapy.

Description

Drug delivery system of targeted neutrophil extracellular trap net and preparation method and application thereof Technical Field The invention belongs to the technical field of targeted drug delivery systems, and particularly relates to a drug delivery system of a targeted neutrophil extracellular trap net, a preparation method and application thereof. Background Neutrophils are important innate immune cells of the body that can capture and clear pathogens during the process of combating pathogen invasion by releasing Neutrophil Extracellular Traps (NETs). NETs are reticular structures composed of DNA, histones and various antibacterial proteins, but can be subjected to out-of-regulation excessive release under pathological conditions such as tumors and the like, and show remarkable tumor promotion effect. Research shows that cytokines and growth factors carried by the NETs can remodel the tumor microenvironment and promote invasion and distant metastasis of tumor cells, so that targeted regulation of the formation and elimination of the NETs is considered as a potential strategy for inhibiting tumor proliferation and preventing metastasis of the tumor. Currently, strategies for modulating NETs include primarily inhibiting their production and directly degrading formed NETs. In one aspect, NETs are reduced from the source by inhibiting the activity of key proteins such as peptide acyl arginine deiminase 4, neutrophil elastase, etc., or by interfering with related signaling pathways such as MAPK, cGAS-STING, etc. On the other hand, direct degradation of formed NETs, especially disruption of their DNA backbone, is a direct and effective means. DNase I, a non-specific endonuclease, digests single-or double-stranded DNA, has been shown to significantly impair both NETs-mediated tumor growth and metastasis. However, DNase I has a short half-life in vivo, is prone to inactivation, and has limited clinical applications. In order to improve the stability and targeting, researchers have tried to load DNase I with a nano-carrier or combine it with a targeting peptide to achieve efficient and accurate degradation of the nes. Recent studies have further revealed the molecular mechanisms by which NETs promote tumor metastasis. For example, the transmembrane protein CCDC25 on the surface of cancer cells is proved to be a specific receptor of NETs-DNA, and can activate downstream ILK-beta-parvin and other signal channels after being combined, induce cytoskeletal remodeling and enhance migration, adhesion and proliferation capacity of tumor cells. Thus, blocking CCDC25 interactions with nes-DNA or specific clearance of nes using CCDC 25-mediated targeted drug delivery systems has become a research hotspot for inhibiting tumor metastasis. Although the prior art has been directed to inhibition, degradation of NETs and CCDC25 targeting strategies, respectively, there is still a lack of a systemic delivery platform capable of integrating multiple mechanisms, synergistically exerting anti-tumor and anti-metastatic effects. Particularly, the multifunctional nano preparation which combines chemotherapy drugs and NETs degradation with CCDC25 mediated active targeting function and simultaneously endows the system with magnetic response and photothermal treatment capability has not been reported at present. Therefore, a composite nano drug delivery system which can realize high-efficiency load, targeted delivery and controllable release in a tumor microenvironment is developed, and has important significance for realizing the synergy of tumor treatment and transfer prevention and control. Disclosure of Invention The invention aims to provide a composite nano drug delivery system with an active targeting function and capable of cooperatively inhibiting tumor growth and metastasis and a preparation method thereof, wherein the composite nano drug delivery system integrates various mechanisms such as magnetic targeting, photothermal therapy, chemotherapy, NETs degradation and the like through a multifunctional integrated design, and aims to improve the anti-tumor curative effect and reduce the toxic and side effects of the whole body. According to the technical scheme, the drug delivery system of the targeted neutrophil extracellular trap net adopts superparamagnetic ferroferric oxide nanoparticles as cores, mesoporous polydopamine shell layers are constructed on the surfaces of the superparamagnetic ferroferric oxide nanoparticles to form a nano carrier Fe 3O4 @mPDA of a core-shell structure, the nano carrier Fe 3O4 @mPDA is used for efficiently co-loading a drug molecule chemotherapeutic drug DOX and degrading DNase I of the neutrophil extracellular trap net to obtain drug-carrying nanoparticles, and the surfaces of the drug-carrying nanoparticles are modified to specifically identify and combine with targeting molecules CCDC25 of the neutrophil extracellular trap net, so that the drug delivery system of the targeted neutrophil extra