CN111388670A

CN111388670A - Synthesis method of mineralized Prussian blue-light immune preparation based on lipopolysaccharide

Info

Publication number: CN111388670A
Application number: CN202010203820.6A
Authority: CN
Inventors: 郑斌; 王树超; 明东; 刘爽; 甘霖
Original assignee: Tianjin University
Current assignee: Tianjin University
Priority date: 2020-03-20
Filing date: 2020-03-20
Publication date: 2020-07-10

Abstract

The invention relates to a method for synthesizing a mineralized Prussian blue-light immune preparation based on lipopolysaccharide, which comprises the steps of 1) weighing and preparing 1.0mM ferric trichloride hexahydrate aqueous solution and 1.0mM potassium ferrocyanide aqueous solution, 2)) and utilizing normal-temperature biomimetic synthesis of Prussian blue-light immune agent (L PS @ PB) nanoparticles, L PS can efficiently activate M1 type macrophages, further induce a large amount of inflammatory factors to activate a whole immune system and cause immune response, and the Prussian blue nanoparticle-guided MRI imaging guidance realizes light-operated release of tumor antigens, further enhances ICD effect and efficiently kills tumors.

Description

Synthesis method of mineralized Prussian blue-light immune preparation based on lipopolysaccharide

Technical Field

The invention relates to the field of preparation of Prussian blue-light immunizing agents, and in particular relates to a synthetic method for biomimetically synthesizing a Prussian blue-light immunizing agent based on lipopolysaccharide.

Background

Malignant tumor (cancer) has become one of the major public health problems seriously threatening the health of Chinese population, and the incidence of malignant tumor is continuously increasing in recent ten years according to the latest statistical data, and the prevention and control situation is increasingly severe. Therefore, the development cost is low and the response measures with good effect are not slow enough.

Recently, immunotherapy for tumor has been developed, which refers to a new direction of tumor therapy that applies immunological principles and methods to increase immunogenicity of tumor cells and sensitivity to killing by effector cells, stimulate and enhance anti-tumor immune response of the body, and deliver immune cells or effector molecules into the body of a patient, in conjunction with the killing of tumor cells by the immune system of the body, and inhibit tumor growth, with few side effects to the human body, L PS is a complex of lipid and polysaccharide, a chemical component unique to the outer wall of gram-negative bacteria, having a molecular weight greater than 10000, consisting of core polysaccharide, O-polysaccharide side chain, and lipid a, when cells are stimulated by L PS, L PS activates nuclear transcription factor NF- κ B through T L R4 on the surface of macrophages, and is classical, thus producing inflammation with the expression of cytokines, including TNT- α, I L-6, and I L-1 α, etc., and can highly express Inducible Nitric Oxide Synthase (iNOS), natural immunity, which can be used as an immune preparation for tumor therapy.

Photothermal therapy (PTT) is a diagnosis and treatment method accompanied by light, has the advantages of minimal invasion, high efficiency, good specificity, space-time controllability and the like, and is an ideal method for treating cancers. During treatment, it can destroy cell membranes within minutes using photothermal conversion, thereby killing cancer cells. 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.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides a Prussian blue-light immunizing agent (L PS @ PB) biomimetically synthesized based on lipopolysaccharide.

A synthetic method for biomimetically synthesizing a Prussian blue-light immunizing agent (L PS @ PB) based on lipopolysaccharide comprises the following steps:

1) 13.5mg of copper chloride dihydrate was weighed, and dissolved by ultrasonic waves after adding 50m L ultrapure water to obtain an aqueous ferric chloride hexahydrate solution having a concentration of 1.0 to 2.0 mM.

2) 21.1mg of gadolinium chloride hexahydrate was weighed, and dissolved by ultrasonic waves after adding 50m L ultrapure water to obtain a 1.0-2.0mM potassium ferrocyanide aqueous solution.

3) The method for biomimetically synthesizing L PS @ PB nanoparticles at normal temperature comprises the following steps:

(1) measuring 20m L ferric chloride hexahydrate solution on a temperature control stirring table, stirring and heating at 60 DEG C

(2) Measuring 20m of L potassium ferrocyanide solution, placing on a temperature-controlled stirring table, stirring and heating at 60 DEG C

(3) After 10 minutes, respectively adding 2-6mg of L PS into the solution, and reacting for 1 hour;

(4) adding a reaction solution of ferric trichloride hexahydrate and L PS into a reaction solution of potassium ferrocyanide and L PS by using a pipette, and reacting for 30 min;

(5) and then cooling the system to room temperature, washing and purifying the system for three times by using ethanol and deionized water, and re-dissolving the precipitate in the deionized water for later use.

The prepared medicine has the following advantages that 1) L PS can efficiently activate M1 type macrophages, further induce a large amount of inflammatory factors to activate the whole immune system and cause immune response, and 2) Prussian blue nanoparticle-guided MRI imaging visualization guidance realizes the light-operated release of tumor antigens, further enhances the ICD effect, induces an organism to generate an immunological memory function, and achieves the purpose of efficiently and thoroughly treating tumors.

Detailed Description

The present invention will be further described below.

Example 1:

1) 13.5mg of copper chloride dihydrate was weighed, and 50m of L mM ultrapure water was added thereto and dissolved by ultrasonic waves to obtain a 1.0mM aqueous solution of ferric chloride hexahydrate.

2) 21.1mg of gadolinium chloride hexahydrate was weighed, and dissolved by ultrasonic waves after adding 50m L ultrapure water to obtain a 1.0mM potassium ferrocyanide aqueous solution.

(1) the ferric chloride hexahydrate solution with the concentration of 20m L was weighed on a temperature-controlled stirring table and stirred and heated at 60 ℃.

(2) A20 m potassium ferrocyanide solution with the concentration of L is weighed and placed on a temperature-controlled stirring table, and stirred and heated at the temperature of 60 ℃.

(3)3) after 10 minutes, 2mg of L PS was added to the above solutions, respectively, and reacted for 1 hour.

(4) Adding the reaction solution of ferric trichloride hexahydrate and L PS into the reaction solution of potassium ferrocyanide and L PS by using a pipette gun, and reacting for 30 min.

Example 2:

1) 27mg of copper chloride dihydrate was weighed, and 50m of L mM ultrapure water was added thereto and dissolved by ultrasonic waves to obtain a 2.0mM aqueous solution of ferric chloride hexahydrate.

2) 42.2mg of gadolinium chloride hexahydrate was weighed, and dissolved by ultrasonic waves after adding 50m L ultrapure water to obtain a potassium ferrocyanide aqueous solution having a concentration of 2.0 mM.

(3) After 10 minutes, 4mg of L PS was added to each of the above solutions and reacted for 1 hour.

Example 3:

1) 40.5mg of copper chloride dihydrate was weighed, and 50m of L mM ultrapure water was added thereto and dissolved by ultrasonic waves to obtain a 1.0mM ferric trichloride hexahydrate aqueous solution.

2) 63.3mg of gadolinium chloride hexahydrate was weighed, and dissolved by ultrasonic waves after adding 50m L ultrapure water to obtain a 1.0mM potassium ferrocyanide aqueous solution.

(3) After 10 minutes, 6mg of L PS was added to each of the above solutions, and the reaction was carried out for 1 hour.

Claims

1. The method for synthesizing the mineralized Prussian blue-light immune preparation based on lipopolysaccharide is characterized by comprising the following steps of:

1) weighing copper chloride dihydrate, adding ultrapure water, and performing ultrasonic dissolution to obtain ferric chloride hexahydrate aqueous solution with the concentration of 1.0-2.0 mM;

2) weighing gadolinium chloride hexahydrate, adding ultrapure water, and performing ultrasonic dissolution to obtain a potassium ferrocyanide aqueous solution with the concentration of 1.0-2.0 mM;

(1) measuring 20m L ferric chloride hexahydrate solution on a temperature control stirring table, and stirring and heating at 60 ℃;

(2) weighing 20m L potassium ferrocyanide solution, placing on a temperature-controlled stirring table, and stirring and heating at 60 ℃;

(3) after 10 minutes, respectively adding 2-6mg of L PS into the solution for reaction;