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Nanoparticles of calcium carbonate-polydopamine coated drug-loaded bacterial outer membrane vesicles as well as preparation method and application of nanoparticles of calcium carbonate-polydopamine coated drug-loaded bacterial outer membrane vesicles

A polydopamine, outer membrane vesicle technology, applied in the fields of immunology and biomedicine, can solve the problems of reducing tumor immune resistance, easy degradation, low targeted delivery efficiency, etc., to inhibit in situ tumor growth and reduce tumor immune resistance. Effect

Active Publication Date: 2022-04-08
SUN YAT SEN UNIV SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Taking this as a target, TGF-β inhibitors (Ti) can improve the tumor microenvironment and reduce tumor immune resistance, and are widely used in tumor therapy, but there are problems of low delivery efficiency and easy degradation

Method used

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  • Nanoparticles of calcium carbonate-polydopamine coated drug-loaded bacterial outer membrane vesicles as well as preparation method and application of nanoparticles of calcium carbonate-polydopamine coated drug-loaded bacterial outer membrane vesicles
  • Nanoparticles of calcium carbonate-polydopamine coated drug-loaded bacterial outer membrane vesicles as well as preparation method and application of nanoparticles of calcium carbonate-polydopamine coated drug-loaded bacterial outer membrane vesicles
  • Nanoparticles of calcium carbonate-polydopamine coated drug-loaded bacterial outer membrane vesicles as well as preparation method and application of nanoparticles of calcium carbonate-polydopamine coated drug-loaded bacterial outer membrane vesicles

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Embodiment 1

[0044] 1. Preparation of nanoparticles of calcium carbonate-polydopamine-coated drug-loaded bacterial outer membrane vesicles, namely CaCO 3 - Preparation of PDA-OMVs (Ti), the specific steps are as follows:

[0045] Step 1. Preparation of Bacterial Outer Membrane Vesicles, i.e. OMVs

[0046] (1) Escherichia coli BL21 was purchased from the China Microbial Culture Collection Center (CGMCC). After being activated by liquid LB medium, the strain was continuously passaged until the growth of the bacteria was stable (the same time as the inoculum of the bacteria reached the logarithmic phase of growth). Acquisition of bacterial outer membrane vesicles (OMVs).

[0047] (2) Inoculate 50 mL of activated Escherichia coli in 2L of LB liquid medium, shake at 180 rpm at 27°C for 2 h, then add 4 mL of 50 mg / mL IPTG inducer to induce bacteria To produce OMVs, continue to shake for 4 hours to collect the bacterial liquid, centrifuge at 8,000 rpm to collect the supernatant, then use a 0.45...

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Abstract

The invention discloses a preparation method of nanoparticles of calcium carbonate-polydopamine coated drug-loaded bacterial outer membrane vesicles, which comprises the following steps: (1) preparation of OMVs: carrying out continuous subculture on escherichia coli strains until the escherichia coli strains grow stably, then inducing activated escherichia coli in logarithmic phase by using an IPTG inducer to generate OMVs, and carrying out freeze drying on the OMVs to obtain the nanoparticles of the drug-loaded bacterial outer membrane vesicles; (2) preparing OMVs (Ti): forming the OMVs (Ti) from the OMVs separated in the step (1) and a fat-soluble TGF-beta inhibitor through a membrane fusion method under an ice-water bath ultrasonic condition; and (3) preparing CaCO3-PDA-OMVs (Ti): adding the OMVs (Ti), CaCl22H2O, Na2CO3 and dopamine into a Tris-HCl (0.25 mM)-HEPES (25mM) buffer solution, and preparing by a one-pot method. The drug-loaded nanoparticles prepared by the invention have the advantages of no toxicity, high-efficiency delivery of inhibitors, activation of autoimmunity and the like.

Description

technical field [0001] The invention belongs to the technical field of immunology and biomedicine, in particular to a calcium carbonate-polydopamine-coated drug-loaded bacterial outer membrane vesicle nanoparticle, a preparation method and application thereof. Background technique [0002] Bacterial outer membrane vesicles (Outer Membrane Vesicles, OMVs) are Gram-negative bacteria and vesicular nanoparticles produced during growth. These small particles are mainly composed of phospholipids, OM proteins and lipopolysaccharides or low lipopolysaccharides. But also containing periplasmic proteins and cell wall components, these small particles become trapped in the cyst lumen during release. Its diameter is between 20 and 200 nm, and it can achieve effective accumulation in the tumor site through the high permeability and long retention (Enhanced permeability and retention, EPR) effect of solid tumors; secondly, the naturally occurring PAMP molecules (such as lipopolysaccharide...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/51A61K47/46A61K41/00A61K45/06A61P35/00A61P35/04B82Y5/00B82Y20/00B82Y30/00B82Y40/00
CPCY02A50/30
Inventor 梅林唐静张帆李倩倩张锦勰
Owner SUN YAT SEN UNIV SHENZHEN
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