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Multifunctional targeting molecular probe for detecting Treg cells in living tumor microenvironment and application thereof

A technology of tumor microenvironment and targeting molecules, which is applied in the field of multifunctional targeting molecular probes and the preparation of anti-tumor drugs, can solve the problems of large damage to patients, achieve the effects of reducing infiltration, improving patient survival rate, and inhibiting growth

Active Publication Date: 2022-02-08
GUANGDONG GENERAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, Treg cell monitoring at this stage is confirmed by pathological sections, but pathological sections are not only harmful to patients, but also not suitable for long-term monitoring

Method used

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  • Multifunctional targeting molecular probe for detecting Treg cells in living tumor microenvironment and application thereof
  • Multifunctional targeting molecular probe for detecting Treg cells in living tumor microenvironment and application thereof
  • Multifunctional targeting molecular probe for detecting Treg cells in living tumor microenvironment and application thereof

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Experimental program
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Effect test

Embodiment 1

[0034] The preparation of embodiment 1 nanoparticle

[0035] 10mg DSPE-SS-PEG 2000 (Commercially available materials are enough), 10mg / mL 10μL CD25-Cy7, 2mg temozolomide and 10mg / mL 10μL indoleamine 2,3-dioxygenase inhibitor (IDO-inhibitor) were dissolved in 1mL THF, and then the above Add THF to 9mL deionized water with vigorous stirring, stir vigorously for 15min, and then stir overnight to remove residual THF in the solution to obtain target nanoparticles; dialyze target nanoparticles with a 2KDa dialysis membrane for 3 times to remove unwrapped substances , and freeze-dry to obtain the desired targeted molecular probe.

Embodiment 2

[0036] The characterization of embodiment 2 nanoparticles

[0037] The morphology and size of the nanoparticles were detected with a field emission high-resolution 2100F transmission electron microscope (TEM) at an accelerating voltage of 200kV. Such as figure 2 The results shown show that the morphology of the above-mentioned nanoprobes is a uniform spherical shape with a diameter of about 50 nm.

[0038] The size distribution of nanoparticles was measured by a Malvern Zetasizer Nano ZS90 (Malvern, UK) instrument, and the results showed that, with figure 2 The TEM results shown are consistent.

[0039] Use Nicolet 759 type Fourier transform infrared spectrometer (FT-IR) and compress the sample with KBr crystal to detect the infrared spectrum of the sample from 4000 to 500, such as image 3 The results shown show the characteristic peaks of each substance.

[0040]The optical properties of CD25-Cy7 and IR780 were detected by UV-visible absorption (Lambda-35 UV-visible sp...

Embodiment 3

[0042] Example 3 Cell Culture

[0043] GL261 cells were incubated in DMEM medium containing 10% FBS and 1% antibiotics (penicillin-streptomycin, 10000 U / mL) in a 37% humidified atmosphere of 5% CO2. Incubate Treg cells isolated from mice in RPMI-1640 medium containing 10% FBS and 1% antibiotics (penicillin-streptomycin, 10,000 U / mL) at 37°C in a humid environment of 5% carbon dioxide. cultured under the conditions.

[0044] Cell counting assay (CCK-8, Dojindo Laboratories, Kumamoto, Japan) was used to measure cell viability.

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Abstract

The invention relates to the technical field of immunomedicine research, in particular to a multifunctional targeting molecular probe for detecting Treg cells in a living tumor microenvironment based on a photoacoustic imaging technology, and further discloses application of the multifunctional targeting molecular probe in preparation of anti-tumor drugs. According to the multifunctional targeting molecular probe for detecting the Treg cells in the living tumor microenvironment, a nanoprobe capable of responding to a tumor microenvironment is formed in a mode of jointly coating a CD-25 fluorescent antibody, a chemotherapeutic drug and an immune checkpoint inhibitor IDO, under the action of EPR, the nanoprobe can be accumulated at the tumor part, and in the presence of glutathione, the CD-25 fluorescent antibody, the chemotherapeutic drug and the immune checkpoint inhibitor IDO wrapped in the probe can be released, so that Treg cells in the tumor microenvironment can be traced through photoacoustic and fluorescence imaging, the number of the Treg cells in the tumor microenvironment can be quantitatively detected in vivo, growth in tumors can be inhibited, and the survival rate of patients can be increased.

Description

technical field [0001] The invention belongs to the technical field of tumor diagnosis research, and in particular relates to a multifunctional targeting molecular probe for detecting Treg cells in living tumor microenvironments based on photoacoustic imaging technology, and further discloses its use for preparing antitumor drugs. Background technique [0002] Glioma is a diffusely infiltrating, heterogeneous, and lethal tumor. Even with maximal surgical and medical treatment, the treatment of glioblastoma remains a serious thorny problem. According to reports, the median survival period of glioma is less than 2 years, and few patients survive long-term. Moreover, because the boundary of glioma is not obvious, the aggressiveness is strong, and the tumor boundary is blurred and inaccurate, it is easy to lead to incomplete surgical resection and the recurrence rate of patients is high. According to research, the main factor leading to the poor prognosis of GBM is a strong imm...

Claims

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Application Information

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IPC IPC(8): A61K49/00A61K49/22A61K9/51A61K47/24A61K31/4188A61K45/06A61P35/00
CPCA61K49/0058A61K49/0021A61K49/0054A61K49/0093A61K31/4188A61K45/06A61K9/5146A61K49/225A61K49/0002A61P35/00
Inventor 聂立铭曾凡楚黄国家吕静
Owner GUANGDONG GENERAL HOSPITAL
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