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Targeting peptide modified traditional Chinese medicine multi-component exosome-like fusion nanoparticle as well as preparation method and application thereof

A technology of exosomes and targeting peptides, applied in the direction of drug combination, nanotechnology for materials and surface science, nanotechnology, etc., can solve the problems of low oral bioavailability, low solubility, toxic and side effects, etc., and achieve improvement Low loading, highly endogenous, improved targeting effect

Active Publication Date: 2022-01-11
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most active ingredients of traditional Chinese medicine have problems such as low solubility, low oral bioavailability, and side effects. For central nervous system diseases, active ingredients of traditional Chinese medicine are difficult to penetrate the BBB

Method used

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  • Targeting peptide modified traditional Chinese medicine multi-component exosome-like fusion nanoparticle as well as preparation method and application thereof
  • Targeting peptide modified traditional Chinese medicine multi-component exosome-like fusion nanoparticle as well as preparation method and application thereof
  • Targeting peptide modified traditional Chinese medicine multi-component exosome-like fusion nanoparticle as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: Extraction and isolation from serum to obtain exosomes (Exo)

[0053] Take blood from the abdominal aorta to obtain rat blood, let it stand for 30min, centrifuge at 1600×g for 12min, and obtain serum; centrifuge at 2000×g for 12min, take supernatant; supernatant 12000×g, centrifuge for 30min, take supernatant liquid, passed through a 0.22 μm aqueous microporous membrane to obtain fresh serum. Add 50% iodixanol solution, 30% iodixanol solution, 10% iodixanol solution, serum, 27000rpm, 4°C, centrifuge for 150min in the centrifuge tube, the exosome density layer is located at 10% and 30% Between the iodixanol solution layers, suck out the exosome density layer into an ultracentrifuge tube, add PBS solution, 40000rpm, 4°C, centrifuge for 80min, and resuspend the obtained precipitate with PBS solution to obtain an exosome (Exo) solution .

Embodiment 2

[0054] Example 2: Preparation process of functionalized liposome (tLipo / CPT) loaded with cryptotanshinone (CPT)

[0055] Take DOPC phospholipid 670μg (850nmol), DOPS phospholipid 300μg (370nmol), DSPE-PEG 2000 -tLyp-1 targeting peptide 230μg (61nmol), CPT powder 100μg (337nmol), dissolved in 1mL of chloroform, fully dissolved, added dropwise to 2mL of PBS buffer, stirring while adding, stirring for 30min, rotary evaporation Remove the organic solvent, transfer the solution to a vial, 195W, and sonicate in an ice-water bath for 5 minutes to obtain tLipo / CPT.

Embodiment 3

[0056] Example 3: Preparation process of exosomes (Exo / SAB) loaded with salvianolic acid B (SAB)

[0057] Take 1 mg of SAB powder and add 21% iodixanol in PBS solution to prepare a 1 mg / mL SAB solution. Take 100 μg of exosome Exo in terms of protein amount, add 40 μL (40 μg) of SAB solution, mix well, add 21% iodixanol PBS solution to make up the total volume to 200 μL. 1000V, 0.5ms, pulse 2 times for electroporation. After electroporation, incubate at a constant temperature of 37°C for 1 hour to promote the closure of the Exo membrane to obtain Exo / SAB.

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Abstract

The invention discloses a preparation method and application of a targeting peptide modified traditional Chinese medicine multi-component exosome-like fusion nanoparticle, the nanoparticle is mainly formed by fusing a functionalized liposome and an exosome, the functionalized liposome is mainly composed of a targeted peptide, phospholipid and a traditional Chinese medicine fat-soluble active drug, the exosome is loaded with a traditional Chinese medicine water-soluble active medicine. The preparation method of the fusion nanoparticles comprises a polyethylene glycol induction method, a repeated freezing and thawing method, an extrusion method, an ultrasonication method and an incubation method. The preparation process is simple, the condition is mild, the cost is low, and the prepared nanoparticles have the advantages of various drug loading types, high penetrability, biological targeting regulation, therapeutic mechanism complementation, biological safety and the like. According to the nano-drug delivery platform, co-delivery, combined administration and collaborative precise treatment of multiple components of traditional Chinese medicine can be achieved, collaborative targeted treatment of complex progressive diseases such as tumors and neurodegenerative diseases can be achieved, and the nano-drug delivery platform has good application prospects.

Description

technical field [0001] The invention belongs to the technical field of nano preparations, and in particular relates to a traditional Chinese medicine multi-component "exosome-like" fusion nanoparticle modified with a targeting peptide, a preparation method and an application method thereof. Background technique [0002] Exosomes are endogenous extracellular vesicles with a diameter of 40-100nm. Various types of cells can secrete exosomes, which are commonly found in body fluids, including blood, urine, saliva, cerebrospinal fluid, etc. . Exosomes contain proteins and RNA related to cell origin, can transport proteins and nucleic acids, participate in intercellular communication, and can be used for disease diagnosis, monitoring, and drug efficacy screening. In addition, the properties of exosomes in delivering active substances to diseased cells promote their research as therapeutic carriers. Exosome membrane surface has a variety of characteristic transmembrane proteins, ...

Claims

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Application Information

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IPC IPC(8): A61K47/69A61K47/64A61K9/51A61K47/46A61K45/06A61P25/28A61P35/00B82Y5/00B82Y30/00B82Y40/00
CPCA61K47/64A61K47/6911A61K47/6901A61K9/5068A61K45/06A61P35/00A61P25/28B82Y5/00B82Y30/00B82Y40/00Y02A50/30
Inventor 狄留庆王若宁赵华聪王雪
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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