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Preparation method and application of a macrophage tracking fluorescent probe

A technology of macrophages and fluorescent probes, applied in the field of preparation of dextran nanoprobes, can solve the problems of renal metabolic clearance, high cost, difficult purification, etc., achieve high biological safety, simple preparation method, and wide application foreground effect

Active Publication Date: 2022-02-22
SHENZHEN INST OF ADVANCED TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Current strategies for targeting tumor-associated macrophages include modifying macrophage-specific ligands and antibodies on the surface of the contrast agent delivery system, such as modifying CD206 antibody or small molecule mannose to target the CD206 protein on the surface of macrophages (also known as mannose receptors), etc. Antibody targeting has good specificity, but purification is difficult and expensive, and small molecule modifications are easily eliminated by renal metabolism.

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  • Preparation method and application of a macrophage tracking fluorescent probe
  • Preparation method and application of a macrophage tracking fluorescent probe
  • Preparation method and application of a macrophage tracking fluorescent probe

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preparation example Construction

[0033] A method for preparing a macrophage tracking fluorescent probe, comprising the following steps:

[0034] S110, placing carboxymethylated dextran, one of NHS and sulfo-NHS, and an activated carboxyl reagent in a buffer solution, stirring at room temperature to obtain an activated carboxymethylated dextran solution.

[0035]Specifically, in one embodiment, the dextran is at least one carboxymethyl dextran with a molecular weight of 2-40 kD and a carboxyl substitution degree of 2%-10%. The activated carboxyl reagent is at least one of EDC, DCC, CDI and DIC. The buffer can be MES buffer or PBS buffer.

[0036] S120, dissolving the cross-linking agent in the buffer solution, adding it into the activated carboxymethylated dextran solution, stirring and reacting at room temperature to obtain a clear solution.

[0037] Specifically, in one embodiment, the cross-linking agent is lysine.

[0038] S130, adding the clarified solution dropwise to pre-cooled absolute ethanol, cent...

Embodiment 1

[0057] 1. Accurately weigh 0.55g of carboxymethylated dextran, add 2.4g of EDC and 0.4572g of NHS, dissolve in 6.2mL of MES buffer (50mM, pH 6.0-6.5), and react with gentle stirring at room temperature for 10 minutes.

[0058] 2. Accurately weigh 0.4g of L-lysine, dissolve it with 0.7mL MES buffer solution (50mM, pH 6.0-6.5), add it to the activated carboxymethylated dextran solution in step 1, and stir gently at room temperature 5h.

[0059] 3. Add the clear solution obtained in step 2 dropwise to 30mL pre-cooled absolute ethanol, centrifuge (2.5k×g, 3min) to collect the white precipitate, redissolve the obtained white precipitate in water, and pass through 0.22μm filter membrane.

[0060] 4. Use ultrapure water as the dialysis medium to perform room temperature dialysis (3 days) on the solution obtained in step 3 with a 10kD dialysis bag. After dialysis, pass through a 0.22 μm filter membrane, pre-freeze in a -20 degree refrigerator for 2 hours, and transfer to -80 degrees ...

Embodiment 2

[0065] 1. Accurately weigh 0.11g of carboxymethylated dextran, add 0.48g of EDC and 0.09144g of NHS, dissolve in 1.24mL of MES buffer (50mM, pH 6.0-6.5), and stir gently at room temperature for 10 minutes.

[0066] 2. Accurately weigh 0.08g of L-lysine, dissolve it with 0.14mL MES buffer solution (50mM, pH 6.0-6.5), add it to the activated carboxymethylated dextran solution in step 1, and stir gently at room temperature 5h.

[0067] 3. Add the clear solution obtained in step 2 dropwise to 6mL pre-cooled absolute ethanol, centrifuge (2.5k×g, 3min) to collect the white precipitate, redissolve the obtained white precipitate in water, and pass through 0.22μm filter membrane.

[0068] 4. Use ultrapure water as the dialysis medium to perform room temperature dialysis (3 days) on the solution obtained in step 3 with a 10kD dialysis bag. After dialysis, pass through a 0.22 μm filter membrane, pre-freeze in a -20 degree refrigerator for 2 hours, and transfer to -80 degrees Refrigerat...

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Abstract

The invention relates to a preparation method and application of a fluorescent probe for macrophage tracing. The probe chemically cross-links carboxymethyl dextran with lysine, one of the essential amino acids for human body, to form uniform and stable dextran. Glycan cross-linked nanoparticles, on which small imaging molecules such as indocyanine green were covalently linked, constructed nanoparticles with both macrophage targeting and imaging capabilities. The probe prepared by the invention has better macrophage targeting ability and higher biological safety.

Description

technical field [0001] The invention relates to the field of material synthesis and application, in particular to a preparation method and application of a dextran nanoprobe targeting macrophages. Background technique [0002] Malignant tumors are a type of disease that seriously threatens the health of residents. According to the latest statistics, the death toll of malignant tumors accounts for 23.91% of the total deaths of residents in my country, and the incidence and death of malignant tumors have continued to rise in the past ten years. The medical expenses caused by tumors exceed 220 billion, and the prevention and control situation is severe. It is urgent to develop real-time, quantitative and sensitive tumor monitoring probes. It is a common idea to directly target and monitor tumor cells, and the tumor area is not only tumor cells, but also infiltrated with many immune cells that regulate tumor progression, forming a tumor stromal environment that can regulate tissu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K49/00C09K11/06C08B37/02
CPCC09K11/06C08B37/0009C08B37/0024A61K49/0034A61K49/0054A61K49/0093
Inventor 郑海荣胡德红罗新平盛宗海马腾高笃阳
Owner SHENZHEN INST OF ADVANCED TECH
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