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Fluorescence contrast agent based on tumor sentinel lymph node detection and preparation method thereof

A fluorescent contrast agent, sentinel lymph node technology, applied in the preparations for in vivo experiments, luminescence/biological staining preparations, pharmaceutical formulations, etc., to achieve the effects of low tissue autofluorescence, low scattering, and improved water solubility and biocompatibility

Active Publication Date: 2021-05-11
山西白求恩医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, combining cuttlefish melanin with near-infrared region II dyes to obtain nanoprobes for the detection of tumor sentinel lymph nodes has not yet been reported.

Method used

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  • Fluorescence contrast agent based on tumor sentinel lymph node detection and preparation method thereof
  • Fluorescence contrast agent based on tumor sentinel lymph node detection and preparation method thereof
  • Fluorescence contrast agent based on tumor sentinel lymph node detection and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0058] Take the freshly dissected cuttlefish ink sac, rinse the ink suspension in the ink sac with water and collect it, add deionized water to wash, centrifuge at 8000 rpm for 15 minutes, collect dark brown precipitate, add absolute ethanol to wash, centrifuge under the same conditions, collect black Brown precipitate. Repeat the above cleaning steps for 6 times, finally wash with deionized water, and freeze-dry to obtain cuttlefish melanin.

[0059] Weigh 6mg of cuttlefish melanin into 6mL of deionized water, and ultrasonically disperse to obtain a 1mg / mL cuttlefish melanin suspension, and adjust the pH to 9.

[0060] Weigh 30 mg of amino-polyethylene glycol-amino (number average molecular weight 5 KDa) and dissolve it in 6 mL of water to prepare a 5 mg / mL aqueous solution, and adjust the pH to 9.

[0061] The above aqueous solution was added dropwise to the cuttlefish melanin suspension, stirred for 18 hours, washed with deionized water, and centrifuged in a 30KD ultrafilt...

Embodiment 2

[0069] Take the freshly dissected cuttlefish ink sac, wash it with water to collect the ink suspension in the ink sac, wash it with deionized water, centrifuge at 10000rpm for 15min, collect the dark brown precipitate, then add absolute ethanol to wash, and collect the dark brown precipitate by centrifugation under the same conditions. Repeat the above cleaning steps for 6 times, finally wash with deionized water, and freeze-dry to obtain cuttlefish melanin.

[0070] Weigh 6mg of cuttlefish melanin into 6mL of deionized water, and ultrasonically disperse to obtain a 1mg / mL cuttlefish melanin suspension, and adjust the pH to 9.5.

[0071] Weigh 20 mg of amino-polyethylene glycol-mercapto (number average molecular weight 10 KDa) and dissolve it in 5 mL of water to prepare a 4 mg / mL aqueous solution, and adjust the pH to 9.5.

[0072] The above aqueous solution was added dropwise to the cuttlefish melanin suspension, stirred for 15 hours, washed with deionized water, and centrifu...

Embodiment 3

[0076] Take the freshly dissected cuttlefish ink sac, wash it with water to collect the ink suspension in the ink sac, wash it with deionized water, centrifuge at 12000rpm for 15min, collect the dark brown precipitate, add absolute ethanol to wash, and collect the dark brown precipitate by centrifugation under the same conditions. Repeat the above cleaning steps for 6 times, finally wash with deionized water, and freeze-dry to obtain cuttlefish melanin.

[0077] Weigh 6mg of cuttlefish melanin into 6mL of deionized water, and ultrasonically disperse to obtain a 1mg / mL cuttlefish melanin suspension, and adjust the pH to 10.

[0078] Weigh 15 mg of amino-polyethylene glycol-amino (number average molecular weight 2KDa) and 15 mg of amino-polyethylene glycol-mercapto (number average molecular weight 2KDa) and dissolve them in 3 mL of water to prepare a 10 mg / mL aqueous solution, and adjust the pH to 10.

[0079] The above aqueous solution was added dropwise to the cuttlefish mela...

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Abstract

The invention discloses a fluorescent contrast agent based on tumor sentinel lymph node detection. The fluorescent contrast agent is prepared by using cuttlefish melanin as a carrier, modifying the cuttlefish melanin with amino polyethylene glycol, and chemically combining the modified cuttlefish melanin with a near-infrared two-region fluorescent dye through an amido bond, wherein the loading capacity of the near-infrared two-region fluorescent dye is 9-18wt%. The fluorescent contrast agent prepared by the invention is applied as a tumor sentinel lymph node contrast agent, has the advantages of high penetration depth, high sensitivity, high signal-to-noise ratio, strong lymph node targeting property, good biological safety and the like, and can meet the requirement of accurate positioning imaging of in-vivo lymph nodes.

Description

technical field [0001] The invention belongs to the technical field of medical diagnostic imaging, and relates to a fluorescent contrast agent for detecting sentinel lymph nodes of tumors, in particular to a fluorescent contrast agent capable of imaging in the second near-infrared region. Background technique [0002] Cancer seriously threatens the health of all mankind. Cancer metastasis is one of the main causes of death of patients. Among them, lymphatic metastasis is one of the common ways of tumor metastasis, and the sentinel lymph node is defined as the first lymph node of the primary tumor to receive lymphatic drainage. Therefore, accurate diagnosis of lymph node metastasis is very important for formulating treatment strategies and evaluating patient prognosis. [0003] The traditional sentinel lymph node biopsy methods are blue dye method and radionuclide tracer method, but both of them have certain defects that limit their clinical popularization. [0004] Due to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00
CPCA61K49/005A61K49/0054A61K49/0093A61K49/0021
Inventor 张瑞平孙菁华董杰蔡雯雯
Owner 山西白求恩医院