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CN-121987588-A - Drug delivery system for relieving autism extracellular exosome as well as preparation method and application thereof

CN121987588ACN 121987588 ACN121987588 ACN 121987588ACN-121987588-A

Abstract

The application discloses a drug delivery system for relieving autism extracellular fluid, a preparation method and application thereof. The system takes a hybrid nano vesicle and oldenlandia diffusa exosome complex as a carrier to load dopamine drugs. The hybrid nanometer vesicle is formed by fusing mesenchymal stem cell exosomes with liposome, and the oldenlandia diffusa exosome compound consists of oldenlandia diffusa exosomes and chitosan. The application also provides a preparation method of the delivery system, which comprises the steps of carrier construction, drug loading and purification. The delivery system has excellent biocompatibility, stability and blood brain barrier penetrating capacity, and can realize targeted delivery and slow release of dopamine. Animal experiments show that the system can obviously improve social behavior of an autism model mouse, reduce the notch plate behavior and relieve anxiety, and has good treatment effect. The application provides a novel delivery strategy which is efficient, safe and can be industrialized for the drug treatment of autism.

Inventors

  • MA YUCHENG
  • Meng Xihong
  • LI HUAN
  • WANG AOWEI
  • WANG FENGBO

Assignees

  • 河北干细胞智慧医疗科技集团有限公司

Dates

Publication Date
20260508
Application Date
20260104

Claims (10)

  1. 1. The drug delivery system for relieving autism cell exosome is characterized in that the drug delivery system takes a hybrid nanovesicle and oldenlandia diffusa exosome complex as a carrier to load dopamine drugs, wherein; The hybrid nanovesicles are fusion of mesenchymal stem cell exosomes and liposomes; the hedyotis diffusa exosome compound consists of a hedyotis diffusa exosome and chitosan.
  2. 2. The autism relieving extracellular exosome drug delivery system according to claim 1, wherein the mass ratio of the hybrid nanovesicles to the oldenlandia diffusa exosome complex is 1-2:1.
  3. 3. The autism relieving extracellular exosome drug delivery system according to claim 1, wherein the mass ratio of mesenchymal stem cell exosome to liposome is 1:1-4.
  4. 4. The autism-alleviating extracellular urinary drug delivery system of claim 1, wherein the liposomes are cationic and anionic liposomes.
  5. 5. The autism relieving extracellular exosome drug delivery system according to claim 1, wherein a mass ratio of the oldenlandia diffusa exosome to the chitosan is 1:5-10.
  6. 6. A method of preparing an autism-alleviating extracellular exosome drug delivery system, the method comprising: mixing the hybrid nanovesicles and the oldenlandia diffusa exosome compound according to a mass ratio of 1-2:1, and gently shaking for 2 hours at 4 ℃ to enable the two compounds to fully interact to form a carrier; dissolving dopamine in PBS with pH of 7.4-7.6 to obtain dopamine solution; and (3) adding the dopamine into the dopamine solution obtained in the step (2) into the carrier obtained in the step (1), and carrying out light shaking overnight at 4 ℃ to obtain the final carrier loaded dopamine medicament.
  7. 7. The method for preparing a drug delivery system for relieving autism according to claim 6, wherein the method comprises the steps of homogenizing oldenlandia diffusa, removing cell debris and large particle impurities through low-speed centrifugation, filtering supernatant with a 0.22 μm filter membrane, centrifuging 70 min at a temperature of 110000xg by using an ultracentrifuge, collecting precipitate to obtain crude extract, re-suspending the crude extract in PBS buffer solution, concentrating and eluting by using an ultrafiltration centrifuge tube to obtain oldenlandia diffusa exosome, mixing the oldenlandia diffusa exosome with chitosan solution (0.1% w/v), and stirring with a gentle magnetic force for 1h according to a mass ratio of 1:5-10, so that chitosan is on the exosome surface to obtain the oldenlandia diffusa exosome compound.
  8. 8. The method of preparing an autism cell exosome drug delivery system according to claim 7, wherein the ultrafiltration centrifuge tube has a molecular weight cut-off of 100 kDa.
  9. 9. The preparation method of the drug delivery system for relieving autism extracellular fluid of claim 6, wherein the preparation method of the hybrid nanovesicles is characterized in that the cultured mesenchymal stem cells are subjected to exosome extraction to obtain mesenchymal stem cell exosomes, phosphatidylcholine, dioleoyl trimethylammonium propane and cholesterol are dissolved, rotary evaporation is carried out, hydration is carried out, liposome water bath ultrasound is carried out to obtain liposomes, and the mesenchymal stem cell exosomes and the liposomes are proportionally frozen and thawed repeatedly to obtain the hybrid nanovesicles.
  10. 10. The delivery system and method of making same according to any one of claims 1-9, wherein the delivery system is used in an autism-alleviating drug.

Description

Drug delivery system for relieving autism extracellular exosome as well as preparation method and application thereof Technical Field The application belongs to the technical field of pharmaceutical preparations, and particularly relates to a drug delivery system for relieving autism extracellular secretion, a preparation method and application thereof. Background Autism, also known as Autism Spectrum Disorder (ASD), is a group of neuro-developmental disorders characterized by social communication disorders, repetitive notch behaviors, and stenosis of interest, whose pathogenesis is complex and may be related to many factors such as genetic factors, neurotransmitter imbalance, immune inflammatory responses, brain dysfunctions, etc. At present, the clinical intervention means of autism is mainly rehabilitation training, and the drug treatment can only pointedly relieve partial accompanying symptoms (such as emotional agitation, inattention and the like), lacks specific drugs capable of improving core symptoms from the pathological mechanism level, is easy to degrade in vivo, has low bioavailability, is difficult to accurately target to lesion sites, and can cause systemic side effects at the same time, so that the clinical application effect of the drug is limited. Neurotransmitter disorders are one of the important pathological features of autism, with dysfunction of the dopamine system being particularly critical. Dopamine, which is a core neurotransmitter regulating social behavior, cognitive function and emotional response, is thought to be closely related to the core symptoms such as social communication disorder, repetitive behavior and the like of autism patients because of insufficient secretion or signal transduction in specific areas of the brain. Therefore, supplementing dopamine or regulating dopamine system function is an important direction in the development of autism drugs. The extracellular body is taken as a natural nanoscale vesicle, has good biocompatibility, low immunogenicity and targeting delivery capability, and the membrane structure can protect the loaded medicine from in vivo enzymolysis, and can accurately target the pathological cells through an intercellular recognition mechanism, so that the extracellular body becomes a research hot spot in the field of medicine delivery. The mesenchymal stem cell exosome not only has the delivery advantage of the common exosome, but also can regulate the cell microenvironment and inhibit inflammatory reaction through the carried bioactive molecules, and provides a synergistic effect for the treatment of the neural development disorder disease, while the plant-derived exosome has the characteristics of wide source, low preparation cost, rich natural active ingredients and the like, and the membrane structure can be combined with biological macromolecules to form a stable compound, so that the delivery performance is further optimized. The liposome is used as a classical artificial nano-carrier, has the advantages of controllable structure and high drug carrying capacity, can form hybrid nano-vesicles by fusion with the cell exosomes, combines the biological targeting of the natural exosomes and the drug carrying flexibility of the liposome, and improves the stability and drug carrying efficiency of the carrier. The chitosan is taken as natural cationic polysaccharide, has good biocompatibility and biodegradability, amino in the molecular structure of the chitosan can be combined with negative charge groups on the surface of an exosome membrane to form a charge composite layer, so that the stability of the exosome can be enhanced, the interaction between a carrier and cells can be promoted, and the internalization efficiency of a medicament can be improved. However, the application of the existing extracellular exosome drug delivery system in the treatment of autism still faces many challenges, namely, single-source exosome targeting and drug carrying capacity are limited, requirements of stable drug loading, blood brain barrier penetration and accurate enrichment of lesion parts are difficult to meet simultaneously, the combination stability of a carrier and the drug is insufficient, the drug is easy to release in advance in a body, the treatment effect is reduced, the side effect risk is increased, in addition, the research of a collaborative delivery system designed for the pathological characteristics of the autism is less, and a mature preparation process and application scheme are not formed yet. Therefore, the development of the high-efficiency drug delivery system based on the composite exosome carrier realizes stable loading, accurate targeted delivery and high-efficiency biological utilization of the dopamine drugs by reasonably designing the carrier composition and the preparation process, and simultaneously synergistically adjusts the related pathological microenvironment of the autism, thereby having important significance for impro