PH (potential of hydrogen)-responsive type ultra-sensitive nanometer fluorescent probe and method for preparing same

A nano-fluorescent probe and ultra-sensitive technology are applied in the field of pH-responsive ultra-sensitive nano-fluorescent probes and their preparation, which can solve the problems of poor biodistribution specificity, poor photostability, and high background signal, and achieve improved imaging signal-to-noise ratio, Efficient enrichment, reducing the effect of background signal

Inactive Publication Date: 2018-01-30
XIDIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In summary, the existing problems in the prior art are: conventional organic small molecule fluorescent probes generally have poor biodistribution specificity, high background signal, and poor photostability

Method used

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  • PH (potential of hydrogen)-responsive type ultra-sensitive nanometer fluorescent probe and method for preparing same
  • PH (potential of hydrogen)-responsive type ultra-sensitive nanometer fluorescent probe and method for preparing same
  • PH (potential of hydrogen)-responsive type ultra-sensitive nanometer fluorescent probe and method for preparing same

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preparation example Construction

[0043] Such as figure 1 As shown, the preparation method of the pH-responsive ultrasensitive nanometer fluorescent probe provided by the embodiment of the present invention comprises the following steps:

[0044] S101: The dye material is adsorbed on the PAA molecular chain through the electrostatic interaction between PAA and the positively charged dye, and the PAA after the dye adsorption is self-assembled in isopropanol to form a template;

[0045] S102: The fluorescence of the dye is quenched by aggregation; the exposed carboxyl group on PAA passes through the interaction with Ca 2+ Coordination, calcium phosphate is deposited on the surface of PAA spheres by adjusting the pH and adding phosphate mineralization to form dye-coated nanoparticles;

[0046] The negatively charged matrix material can be calcium phosphate, calcium carbonate, calcium hydroxyphosphate, fluorapatite, etc., and the positively charged dyes used in this example can be IR780, RhB, IR800, etc. Synthes...

Embodiment 1

[0055] Preparation of CaP / IR780 nanoprobe

[0056] Synthesis principle: through the electrostatic interaction between PAA and IR780, the dye is aggregated to achieve fluorescence quenching. After the dye is adsorbed, PAA self-assembles in isopropanol to form a template. Calcium phosphate is deposited on the surface of PAA balls through mineralization to form CaP / IR780 nanoparticles.

[0057] Specific synthesis steps: 60-80 μL of 20% PAA (MW=2000) was dissolved in 10 mL of water, and 5-8 mg of Ca(OH) was added 2 Stir to make it dissolve, add 50~1000μg IR780, stir for 30min, and make it fully mixed. Slowly add 10~50mL isopropanol to form a milky white solution, add 6~9.6mg (NH 4 ) 2 HPO 4 Stir for 24 hours, centrifuge, wash 3 times with water, redisperse in 5mL water for later use. Particle size characterization by transmission electron microscopy (TEM), scanning electron microscopy (SEM) and dynamic light scattering (DLS), such as figure 2 , image 3 and Figure 4 sho...

Embodiment example 2

[0059] Preparation of CaP / RhB Nanoprobes

[0060] Synthesis principle: The dye is aggregated through the electrostatic interaction between PAA and RhB, thereby realizing fluorescence quenching. The PAA after the dye is self-assembled in isopropanol to form a template, and calcium phosphate is deposited on the surface of the PAA ball through mineralization, thereby forming CaP / RhB nanoparticles.

[0061] Specific synthesis steps: 60-90 μL of 20% PAA (MW=4500) was dissolved in 10 mL of water, and 5-10 mg of Ca(OH) was added 2 Stir to dissolve, add 100-1000μg RhB, stir for 30min, and make it fully mixed. Slowly add 30mL of isopropanol to form a milky white solution, add 6.2~12mg (NH 4 ) 2 HPO 4 Stir for 24 hours, centrifuge, wash with water 3 times, and redisperse in 5mL of water. In order to improve the stability of the nanoprobe and the circulation time in the animal blood, the surface of the probe is stabilized by PEG, thereby improving the stability of the probe. Add 5...

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Abstract

The invention belongs to the field of molecular imaging technologies, and discloses a pH (potential of hydrogen)-responsive type ultra-sensitive nanometer fluorescent probe and a method for preparingthe same. The pH-responsive type ultra-sensitive nanometer fluorescent probe comprises pH-responsive matrix materials and fluorescent organic small-molecule dye. The pH-responsive matrix materials comprise calcium phosphate, hydroxy calcium phosphate, fluorapatite, calcium carbonate and ZIF series; the fluorescent organic small-molecule dye is positively charged dye or negatively charged dye. Themethod includes coating the positively charged dye with the negatively charged matrix materials; coating the negatively charged dye with the negatively charged matrix materials; coating the negativelycharged dye with the positively charged matrix materials. The pH-responsive type ultra-sensitive nanometer fluorescent probe and the method have the advantages that the fluorescent imaging sensitivity and specificity can be greatly improved as compared with the traditional small-molecule fluorescent dye, and response of tumor micro-environments can be detected in an ultra-sensitive manner; the pH-responsive type ultra-sensitive nanometer fluorescent probe which is a specific responsive probe prepared for unique properties of the tumor micro-environments is high in targeting, few in backgroundsignals and high in signal-to-noise ratio, small tumor can be detected in an ultra-sensitive manner, and the like.

Description

technical field [0001] The invention belongs to the technical field of molecular imaging, and in particular relates to a pH-responsive ultra-sensitive nanometer fluorescent probe and a preparation method thereof. Background technique [0002] Molecular imaging technology can be used to study the occurrence, development and metastasis of diseases or tumors in vivo because it can realize real-time, non-invasive and dynamic imaging at the living level. Optical imaging technology has the advantages of no radiation, high sensitivity, and less photodamage to biological tissues, and has shown great potential in the field of tumor monitoring in vivo. Due to the rapid proliferation of cells in the tumor tissue, the glucose metabolism is fast, the oxygen consumption is large, and the acidic metabolites (such as lactic acid) accumulate in the tumor microenvironment. Therefore, the tumor microenvironment is generally weakly acidic, hypoxic, and produces many cytokines. and high express...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00
CPCA61K49/0093A61K49/0032A61K49/0034A61K49/0041A61K49/0054
Inventor 王忠良王永东张瑞丽贾茜胥强强乔晁强田捷
Owner XIDIAN UNIV
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