阅读说明：本技术 LPS@CuS:Gd光免疫剂的合成方法 (Method for synthesizing LPS @ CuS Gd photo-immunizing agent ) 是由 郑斌 王树超 明东 刘爽 甘霖 于 2020-03-20 设计创作，主要内容包括：本发明涉及一种通过常温仿生合成成功制备出的LPS@CuS:Gd光免疫剂。包括(1)配置浓度为0.001M的CuCl<Sub>2</Sub>水溶液,0.0025M的GdCl<Sub>3</Sub>水溶液。(2)利用常温仿生合成LPS@CuS:Gd纳米颗粒。其中LPS可以利用强烈免疫激活功能来活化全身免疫系统,募集DC细胞,引起免疫应答；Gd元素引导的MRI成像可视化引导实现了肿瘤抗原的光控释放,进一步增强了ICD效应,并诱使机体产生免疫记忆功能,达到高效、彻底治疗肿瘤的目的。(The invention relates to an LPS @ CuS and Gd photo-immunizing agent successfully prepared by normal-temperature biomimetic synthesis. Comprises (1) preparing CuCl with the concentration of 0.001M 2 Aqueous solution, 0.0025M GdCl 3 An aqueous solution. (2) LPS @ CuS and Gd nanoparticles are biomimetically synthesized at normal temperature. The LPS can activate the whole body immune system by utilizing the strong immune activation function, recruit DC cells and cause immune response; gd (Gd)The element-guided MRI imaging visual guidance realizes the light-operated release of tumor antigens, further enhances the ICD effect, induces the organism to generate the immunological memory function and achieves the aim of efficiently and thoroughly treating tumors.)

The method for synthesizing the LPS @ CuS Gd photo-immunizing agent is characterized by comprising the following steps of:

1) weighing copper chloride dihydrate, adding ultrapure water, and ultrasonically dissolving to obtain CuCl with concentration of 0.001-0.004M₂An aqueous solution;

2) weighing gadolinium chloride hexahydrate, adding 10mL of ultrapure water, and ultrasonically dissolving to obtain GdCl with concentration of 0.0025-0.01M₃An aqueous solution;

3) the method for synthesizing the LPS @ CuS/Gd nano-particles by biomimetic mineralization comprises the following steps:

(1) accurately weighing 0.25-1mg of lipopolysaccharide, adding into a 5mL transparent bottle, adding 5mL of ultrapure water, and ultrasonically dissolving until the powder is fully dissolved to obtain an LPS aqueous solution with the concentration of 0.5-2 mg/mL;

(2) placing 1ml LPS solution single-mouth bottle on 37 deg.C water bath magnetic stirrer, and dripping 5ml CuCl prepared above with disposable dropper under stirring₂And 1ml GdCl₃Continuously stirring the solution for three minutes after the dropwise addition;

(3) preparing 0.01M NaOH solution, adding 0.5mL of NaOH solution into the reaction system, and adjusting the pH value of the solution to 10;

(4) preparing sodium sulfide nonahydrate solution with the concentration of 0.01-0.03mmol/mL, and adding 0.04mL into a reaction system;

(5) stirring at 37 deg.C for 3 hr, taking out reaction solution, centrifuging at 10000 rpm, collecting precipitate, repeatedly adding deionized water, cleaning for 3 times, and storing at 4 deg.C.

Technical Field

The invention relates to the technical field of photo-immune preparations, in particular to a method for synthesizing an LPS @ CuS/Gd photo-immune agent through normal-temperature biomimetic synthesis.

Background

Recently, tumor immunotherapy has become an important breakthrough and development direction for tumor therapy. Different from the traditional treatment mode, the tumor immunotherapy mainly achieves the aim of treating tumors by activating or improving the immunologic function of a human body, and has small side effect on the human body; meanwhile, immunotherapy has a long-term anti-tumor immune function and can specifically identify tumor cells, so that the metastasis and recurrence of tumors are effectively inhibited. LPS is a compound of lipid and polysaccharide, is a specific chemical component in the outer wall layer of gram-negative bacteria, and has molecular weight of more than 10000. It consists of core polysaccharide, O-polysaccharide side chains, and lipid A. It is a natural ligand for TOLL-like receptor 4(TLR4) and induces DC maturation and a strong TH 1-type immune response. It is therefore intended herein to be used as an immunostimulant to effect immunotherapy.

The photothermal therapy (PTT) is a novel tumor therapy method, under the irradiation of external near infrared light, a photothermal agent at a tumor part absorbs the near infrared light and converts the near infrared light into heat, so that the temperature of the tumor part is rapidly increased to more than 48 ℃, and cancer cells can be killed within a few minutes. The photothermal therapy process has less side effect, low systemic toxicity, no damage to normal tissue and great clinical application potential. If the photothermal therapy and the tumor immunotherapy are combined, the tumor therapy effect is greatly enhanced, and the tumor therapy is more thorough.

The prepared medicine has the following advantages: 1) LPS can activate the whole body immune system by utilizing the strong immune activation function, recruit DC cells and cause immune response; 2) the Gd element-guided MRI imaging visual guidance realizes the light-operated release of tumor antigens, further enhances the ICD effect, induces the organism to generate the immunological memory function, and achieves the aim of efficiently and thoroughly treating tumors.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides an LPS @ CuS and Gd photo-immunizing agent successfully prepared by normal-temperature biomimetic synthesis.

The invention relates to a method for synthesizing an LPS @ CuS Gd photo-immunizing agent; the method comprises the following steps: the method comprises the following steps:

1) weighing copper chloride dihydrate, adding ultrapure water, and ultrasonically dissolving to obtain CuCl with concentration of 0.001-0.004M₂An aqueous solution;

2) weighing gadolinium chloride hexahydrate, adding 10mL of ultrapure water, and ultrasonically dissolving to obtain GdCl with concentration of 0.0025-0.01M₃An aqueous solution;

3) the method for synthesizing the LPS @ CuS/Gd nano-particles by biomimetic mineralization comprises the following steps:

(1) accurately weighing 0.25-1mg of lipopolysaccharide, adding into a 5mL transparent bottle, adding 5mL of ultrapure water, and ultrasonically dissolving until the powder is fully dissolved to obtain an LPS aqueous solution with the concentration of 0.5-2 mg/mL;

(2) placing 1ml LPS solution single-mouth bottle on 37 deg.C water bath magnetic stirrer, and dripping 5ml CuCl prepared above with disposable dropper under stirring₂And 1ml GdCl₃Continuously stirring the solution for three minutes after the dropwise addition;

(3) preparing 0.01M NaOH solution, adding 0.5mL of NaOH solution into the reaction system, and adjusting the pH value of the solution to 10;

(4) preparing sodium sulfide nonahydrate solution with the concentration of 0.01-0.03 mmol/mL), and adding 0.04mL into the reaction system;

stirring at 37 deg.C for 3 hr, centrifuging at 10000 rpm, collecting precipitate, repeatedly washing with deionized water for 3 times, and storing at 4 deg.C

The invention has the advantages that: 1) LPS can activate the whole body immune system by utilizing the strong immune activation function, recruit DC cells and cause immune response; 2) the Gd element-guided MRI imaging visual guidance realizes the light-operated release of tumor antigens, further enhances the ICD effect, induces the organism to generate the immunological memory function, and achieves the aim of efficiently and thoroughly treating tumors.

Detailed Description

The present invention will be further described below.