CN111450082A

CN111450082A - Synthesis method of nanoparticles for treating systemic lupus erythematosus

Info

Publication number: CN111450082A
Application number: CN202010340927.5A
Authority: CN
Inventors: 郑斌; 彭文畅; 明东; 甘霖
Original assignee: Tianjin University
Current assignee: Tianjin University
Priority date: 2020-04-26
Filing date: 2020-04-26
Publication date: 2020-07-28

Abstract

The invention discloses a synthesis method of nanoparticles for treating systemic lupus erythematosus, which mainly comprises the steps of 1) preparing PEG aqueous solution, 2) preparing P L GA organic solution, and 3) synthesizing I L-10 @ HIV-Vpu protein nanoparticles by a thin film hydration method, wherein an accessory protein U (Vpu) can inhibit the activation of a transcription factor called NF-kB, reduce the generation of cell factors playing a key role in immune reaction and inhibit the immune function of an organism.

Description

Synthesis method of nanoparticles for treating systemic lupus erythematosus

Technical Field

The invention relates to the technical field of nanoparticle synthesis, in particular to a method for synthesizing an HIV-Vpu protein @ I L-10 @ PEG-P L GA nano preparation by a strategy of wrapping HIV-Vpu protein and interleukin-10 (I L-10) by P L GA-PEG.

Background

Systemic lupus erythematosus (S L E) is involved in complex pathological mechanisms, causing multiple organ damage and potentially life threatening, non-specific immunosuppressive or anti-inflammatory agents such as Glucocorticoids (GCs), cyclophosphamide and methotrexate, are commonly used drugs in S L E however, the efficacy of this therapy is sometimes unsatisfactory and continued use causes serious side effects, which highlights the need to explore a more effective, safe and effective treatment.

Interleukin 10 is a multi-cell-derived, multifunctional cytokine that regulates the growth and differentiation of cells, participates in inflammatory and immune responses, and is a currently recognized inflammatory and immunosuppressive factor. Avoid the excessive production of immune factors in the glomerulus and prevent the damage of the immune system to the glomerulus.

P L GA-PEG is a two-block polymer, in which P L GA is formed by random polymerization of two monomers, lactic acid and glycolic acid, and is a degradable functional high molecular organic compound, its degradation products are lactic acid and glycolic acid, and at the same time, it is a by-product of human metabolic pathway, so that it has no toxic side effect when it is used in medicine and biological material, and has good biocompatibility, non-toxicity, good capsulizing and film-forming properties.

Disclosure of Invention

The invention provides a method for synthesizing a systemic lupus erythematosus treatment nanoparticle preparation by a strategy of wrapping HIV-Vpu protein and interleukin-10 (I L-10) by P L GA-PEG, aiming at overcoming the defects of the prior art, and preventing an immune system from attacking the self by utilizing the HIV-Vpu protein and I L-10 to inhibit the immune overstimulation reaction so as to achieve the purpose of treating the systemic lupus erythematosus.

The technical scheme of the invention is a synthesis method of nanoparticles for treating systemic lupus erythematosus, and the strategy of wrapping HIV-Vpu protein and interleukin-10 (I L-10) by P L GA-PEG comprises the following specific steps:

1) weighing a P L GA material, adding dichloromethane, and performing ultrasonic dissolution to obtain a P L GA organic solution with the concentration of 0.25-1 mg/ml;

2) weighing PEG material, adding water, and ultrasonic dissolving to obtain PEG water solution with concentration of 0.25-1 mg/ml;

3) weighing I L-10 materials, adding dichloromethane, and performing ultrasonic dissolution to obtain an I L-10 organic solution with the concentration of 0.5-2 mg/ml;

4) HIV-Vpu protein @ I L-10 @ PEG-P L GA nanoparticles are synthesized by a thin film hydration method.

(1) Placing a single-mouth bottle containing 2ml of P L GA solution on an ultrasonic crusher, and starting ultrasonic treatment for 12min at 0 ℃ and 4s and 2s at ultrasonic frequency;

(2) adding 100u L I L-10 solution and 2ml PEG water solution drop by drop while carrying out ultrasonic treatment, and continuing ultrasonic treatment until the mixture is completely mixed;

(3) the sonicated product was immediately added to a round bottom flask and rotary evaporated on a rotary evaporator until the dichloromethane and water were all evaporated to dryness and a thin film formed at the bottom of the flask.

(4) Adding 1m L diluted HIV-Vpu protein into a round-bottomed bottle filled with liposome, blowing, suspending and stirring until uniform mixing, and finally obtaining the HIV-Vpu protein @ I L-10 @ PEG-P L GA nanoparticles.

The invention has the advantages that:

1) the HIV-Vpu protein inhibits activation of a transcription factor called NF-kB, reducing the production of cytokines that play a critical role in the immune response.

2) Interleukin-10 (I L-10) Interleukin 10 is a multi-cell derived, multifunctional cytokine that regulates cell growth and differentiation, participates in inflammatory and immune responses, and is currently recognized as an inflammatory and immunosuppressive factor.

3) The P L GA-PEG has good biocompatibility, no toxicity, and good encapsulation and film-forming properties.

Drawings

FIG. 1: nanoparticle TEM images.

Detailed Description

The invention is further described below with reference to the following figures and specific examples.

Example 1:

1) weighing 10mg of P L GA material, adding 20ml of dichloromethane, and performing ultrasonic dissolution to obtain a P L GA organic solution with the concentration of 0.5 mg/ml;

2) weighing 10mg of PEG material, adding 20ml of water, and performing ultrasonic dissolution to obtain a PEG aqueous solution with the concentration of 0.5 mg/ml;

3) weighing 20mg of I L-10 material, adding 20ml of dichloromethane, and performing ultrasonic dissolution to obtain 1mg/ml I L-10 organic solution;

Example 2:

1) weighing 5mg of P L GA material, adding 20ml of dichloromethane, and performing ultrasonic dissolution to obtain a P L GA organic solution with the concentration of 0.25 mg/ml;

2) weighing 5mg of PEG material, adding 20ml of water, and performing ultrasonic dissolution to obtain a PEG aqueous solution with the concentration of 0.25 mg/ml;

3) weighing 10mg of I L-10 material, adding 20ml of dichloromethane, and performing ultrasonic dissolution to obtain an I L-10 organic solution with the concentration of 0.5 mg/ml;

Example 3:

1) weighing 20mg of P L GA material, adding 20ml of dichloromethane, and performing ultrasonic dissolution to obtain a P L GA organic solution with the concentration of 1 mg/ml;

2) weighing 20mg of PEG material, adding 20ml of water, and performing ultrasonic dissolution to obtain 1mg/ml PEG aqueous solution;

3) weighing 30mg of I L-10 material, adding 20ml of dichloromethane, and performing ultrasonic dissolution to obtain 1.5mg/ml I L-10 organic solution;

Claims

1. A synthesis method of nanoparticles for treating systemic lupus erythematosus is characterized in that a strategy of wrapping HIV-Vpu protein and I L-10 by PEG-P L GA comprises the following specific steps:

2. The method for synthesizing nanoparticles for treating systemic lupus erythematosus, according to claim 1, is characterized in that the nanoparticles for inhibiting the immune overstimulation of HIV-Vpu protein @ I L-10 @ PEG-P L GA are prepared, and the strategy of wrapping HIV-Vpu protein and I L-10 by PEG-P L GA is that the step 4) is as follows:

(2) dropwise adding I L-10 solution and 2ml of PEG aqueous solution while performing ultrasonic treatment, and continuing ultrasonic treatment until the mixture is completely mixed;

(3) immediately adding the ultrasonic-treated product into a round-bottomed bottle, and performing rotary evaporation on a rotary evaporator until dichloromethane and water are completely evaporated to dryness to form a film at the bottom of the bottle;