SiRNA-loaded MSR-1 magnetosome compound and preparation method thereof

A MSR-1, magnetosome technology, applied in the directions of non-active ingredients medical preparations, drug combinations, pharmaceutical formulations, etc., can solve problems such as difficulty in passing through the lipid bilayer, unstable siRNA, and short half-life. , to achieve the effect of uniform shape, convenient operation and simple process

Inactive Publication Date: 2017-10-24
HUAQIAO UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The main problems of siRNA delivery in vivo are: (1) siRNA is extremely unstable: due to the presence of ribonuclease, siRNA is easily degraded in vivo, and its half-life is also very short; (2) siRNA needs to go through multiple stages to reach target cells. Obstacles: Since siRNA itself has electronegativity and strong polarity, it is difficult for it to pass through the lipid bilayer, and because it cannot pass through the vascular endothelium quickly, it is easy to stay in blood storage organs such as the liver and spleen, and is easy to be absorbed by the glomerulus of the kidney. (3) siRNA can activate the immune system in vivo to produce immune response and off-target effects
The use of magnetosomes as siRNA drug carriers has not been reported yet

Method used

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  • SiRNA-loaded MSR-1 magnetosome compound and preparation method thereof
  • SiRNA-loaded MSR-1 magnetosome compound and preparation method thereof
  • SiRNA-loaded MSR-1 magnetosome compound and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment

[0027] A method for preparing siRNA-loaded MSR-1 magnetosome complexes, mixing polyethyleneimine (PEI) liquid with siRNA solution (20 μmol / L), and then making magnetosomes (BMs) into suspension with DEPC water solution (0.1mg / mL) was slowly added dropwise to the mixture of polyethyleneimine and siRNA, and the magnetosomes were derived from Magnetospirillumgryphiswaldense MSR-1 magnetotactic bacteria; then vortexed for 2min, and stood at room temperature for 25min to obtain loading MSR-1 magnetosome complexes of siRNA (BMs / PEI / siRNA).

[0028] Wherein, the polyethyleneimine is a branched polyethyleneimine with a molecular weight of 25KD.

experiment example 1

[0029] Experimental Example 1: Preparation of siRNA-loaded MSR-1 magnetosome complexes (BMs / PEI / siRNA) under different N / P ratios (nitrogen-to-phosphorus ratios)

[0030] Referring to the method of the above-mentioned examples, different amounts of PEI were mixed with a certain amount of siRNA so that the N / P ratios were 0, 1, 2, 4, 8, 10, 16, and 20 respectively; then the BMs suspension was slowly added dropwise to In the above-mentioned mixtures of polyethyleneimine and siRNA with different N / P ratios, vortex for 2 minutes, and stand at room temperature for 25 minutes to obtain siRNA-loaded MSR-1 magnetosome complexes (BMs / PEI / siRNA).

[0031] The obtained BMs / PEI / siRNA complex was detected by agarose gel electrophoresis, and the results were as follows: figure 1 shown, from figure 1 It can be seen that when the N / P ratio is small, clear siRNA bands can be observed; as the N / P ratio gradually increases, the bands gradually weaken to no color development, indicating that siR...

experiment example 2

[0033] Experimental Example 2: Preparation of siRNA-loaded MSR-1 magnetosome complexes (BMs / PEI / siRNA) at different BMs / siRNA mass ratios

[0034] Referring to the method of the above example, the N / P ratio is fixed at 20, a certain amount of PEI is mixed with an appropriate amount of siRNA, and then different amounts of magnetosome suspension are slowly added dropwise to the above-mentioned mixture of polyethyleneimine and siRNA, The mass ratios (BMs / siRNA, w / w) of the magnetosomes in the magnetosome suspension to the siRNA in the mixture of polyethyleneimine and siRNA were 1:5, 1:2, 1:1, 2, respectively. :1, 5:1; then vortex for 2 minutes, and stand at room temperature for 25 minutes to obtain the MSR-1 magnetosome complex (BMs / PEI / siRNA) loaded with siRNA.

[0035] The obtained BMs / PEI / siRNA complex was detected by agarose gel electrophoresis, and the results were as follows: Figure 4 shown, from Figure 4 It can be seen that the nanocomposites prepared with different BMs ...

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Abstract

The invention discloses an siRNA-loaded MSR-1 magnetosome compound and a preparation method thereof. The method comprises the steps of mixing polyethyleneimine and siRNA; slowly dropwise adding MSR-1 magnetosome suspension into a mixture of the polyethyleneimine and the siRNA; and carrying out vortex oscillation and standing at a room temperature to obtain the siRNA-loaded MSR-1 magnetosome compound.The MSR-1 magnetosome compound with a natural lipid bilayer membrane is taken as a carrier matrix material, and a negatively charged siRNA molecule is packaged with positive charges of the polyethyleneimine, thereby carrying out electrostatic bonding with a negatively charged magnetosome and building an siRNA-loaded magnetic-targeted anti-cancer drug delivery system. The MSR-1 magnetosome drug carrier prepared through the method has the characteristics of being uniform in form, controllable particle size, relatively narrow in particle size distribution and packaged in biofilm; the siRNA is completely bonded, has serum stability and enzyme-resistant capability, and is obvious in tumor suppression effect; meanwhile, the method is simple in process and convenient to operate; and the MSR-1 magnetosome drug carrier does not relate to a toxic organic solvent and is an environment-friendly drug carrier, so that the siRNA-loaded MSR-1 magnetosome compound has high practicability and a wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of biomedical drug carriers, and in particular relates to an MSR-1 magnetosome complex loaded with siRNA and a preparation method thereof. Background technique [0002] Gene therapy is a focus of current research. Although clinical studies have shown that siRNA gene therapy is effective, most drugs are currently in phase I clinical trials, and few siRNA drugs have entered phase III clinical trials. The problem of in vivo delivery is the biggest problem hindering the development of siRNA drugs. [0003] The main problems of siRNA delivery in vivo are: (1) siRNA is extremely unstable: due to the presence of ribonuclease, siRNA is easily degraded in vivo, and its half-life is also very short; (2) siRNA needs to go through multiple stages to reach target cells. Obstacles: Since siRNA itself has electronegativity and strong polarity, it is difficult for it to pass through the lipid bilayer, and because it canno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/59A61K47/54A61K47/69A61K48/00A61K31/713A61P35/00
Inventor 刘源岗王士斌代晴蕾龙瑞敏
Owner HUAQIAO UNIVERSITY
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