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Homologous recombination exosome multi-drug delivery bionic nanoparticles as well as preparation method and application thereof

A homologous recombination, biomimetic nanotechnology, applied in pharmaceutical formulations, medical preparations without active ingredients, and medical preparations containing active ingredients, etc., can solve the problems of low solubility, low oral bioavailability, toxic and side effects, etc. Achieve high penetration, large drug loading capacity, and reduce toxicity

Active Publication Date: 2022-04-22
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 of the active ingredients of traditional Chinese medicine have problems such as low solubility, low oral bioavailability, and toxic and side effects. For central nervous system diseases, it is difficult for active ingredients of traditional Chinese medicine to pass through the blood-brain barrier to exert their medicinal effects.

Method used

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  • Homologous recombination exosome multi-drug delivery bionic nanoparticles as well as preparation method and application thereof
  • Homologous recombination exosome multi-drug delivery bionic nanoparticles as well as preparation method and application thereof
  • Homologous recombination exosome multi-drug delivery bionic nanoparticles as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1: Extraction and recombination of tumor-derived exosomes

[0052] Replace the serum-containing medium of GL261 cells with serum-free medium, incubate at 37°C for 48h, then centrifuge at 300g for 10min, 2000g for 10min and 10000g for 30min at 4°C, and filter with a 0.22μm water filter membrane Supernatant to remove cells and debris. The filtrate was centrifuged at 120,000 g for 70 min at 4° C. in a Ti70 rotor using an ultracentrifuge. The pellet was resuspended in PBS and ultracentrifuged again at 120,000 g for 70 min, and then the precipitated exosome pellet was resuspended in PBS to obtain tumor-derived exosomes (EXO).

[0053] Sonicate the EXO solution for 5 minutes with a 2s / 5s interval on / off ultrasonic instrument, repeat three times, store at 37°C for 1 hour, then centrifuge the solution at 100,000g for 70 minutes, resuspend and collect in PBS, and obtain homologous recombinant exocrine body (R-EXO).

Embodiment 2

[0054] Example 2: Preparation process of homologous recombinant exosomes (R-EXO-T / D) loaded with temozolomide (TMZ) and dihydrotanshinone (DHT)

[0055] Take 30 mg of TMZ powder and add 1 mL of dimethyl sulfoxide (DMSO) solution to prepare 30 mg / mL TMZ mother solution; take 1 mg of DHT powder and add 1 mL of dimethyl sulfoxide (DMSO) solution to prepare 1 mg / mL DHT mother solution. Take 1 mL of TMZ mother solution and 1 mL of DHT mother solution, add 4 mL of PBS and mix thoroughly to prepare 5 mg / mL TMZ / DHT (T / D) mixed mother solution.

[0056] Investigate the dosage of R-EXO and T / D used in the preparation of R-EXO-T / D, see Table 2-1 for the specific investigation prescription, take an appropriate amount of preparation, and measure the particle size and polydispersity of the preparation with a Malvern laser particle size analyzer coefficient (PDI) and Zeta potential. Draw 200 μl of the prepared R-EXO-T / D solution, add 20 times the volume of methanol, sonicate for 10 minutes,...

Embodiment 3

[0061] Example 3: Investigation of the properties of biomimetic nanoparticles for multi-drug delivery by homologous recombination exosomes

[0062] Select the optimal prescription for the preparation of R-EXO-T / D, and conduct the following investigations:

[0063] 1.1 Particle size change of R-EXO

[0064] Get EXO and R-EXO under the embodiment one, carry out the mensuration of nanoparticle size, the result is as follows figure 1 As shown, the particle size of homologously recombined exosome R-EXO is smaller than that of homologous exosome EXO, which also verifies to a certain extent that the recombination process removes the exosome content, resulting in the exosome particle size decrease.

[0065] 1.2 SDS-PAGE verification of the retention of recombinant exosome membrane proteins

[0066] GL261 cells were stored in PBS solution containing protease inhibitors, ultracentrifuged at 100,000 g for 5 h, and resuspended in PBS to obtain tumor cell membrane (EM). The total prote...

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Abstract

The invention discloses a bionic nanoparticle for homologous recombination exosome multi-drug delivery and a preparation method and application thereof, the bionic nanoparticle is mainly formed by homologous recombination exosome loaded drugs, and the homologous recombination exosome is mainly formed by recombination treatment of exosomes from treatment target cells. The preparation method of the bionic nanoparticles comprises a gradient centrifugation 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 high endogenous property, high targeting property, multi-drug combined delivery, complementary treatment mechanisms, biological safety, diversified treatment means and the like. The bionic nano-drug delivery platform can realize combined delivery of multiple drugs of Chinese and western medicines, realizes collaborative targeted treatment of complex progressive diseases such as tumors and neurodegenerative diseases, and has a good application prospect.

Description

technical field [0001] The invention belongs to the technical field of nano preparations, and in particular relates to a biomimetic nanoparticle for homologous recombination exosome multi-drug delivery and its preparation method and application. Background technique [0002] Exosomes are microvesicles with a diameter of 30-150 nm, which are collectively called extracellular microvesicles together with microvesicles and apoptotic bodies. Exosomes can not only mediate intercellular communication through proteins, lipids, DNA, and mRNA, but also serve as carriers of small molecule drugs or nucleic acid drugs to deliver therapeutic molecules to specific target sites for targeted delivery. Therefore, exosomes, as a natural drug delivery system, are being widely used in the precise delivery of drugs. In contrast to synthetic carriers, endogenous drug delivery vehicles are more compatible with the complex in vivo environment and can home to target tissues more efficiently, thus pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/50A61K47/46A61K45/00A61K31/4188A61P35/00A61P25/00
CPCA61K9/5068A61K45/00A61K31/4188A61P35/00A61P25/00
Inventor 王若宁梁启凡张新茹孙丹妮狄留庆
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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