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Application of water-soluble cationic porphyrin in preparation of PDT nano photosensitizer

A technology of water-soluble cations and photosensitizers, used in biochemical equipment and methods, microbial determination/inspection, wave energy or particle radiation treatment materials, etc., can solve the problems of low dark toxicity and poor biocompatibility, and achieve high light Toxicity, low cost, easy metabolic decomposition effect

Active Publication Date: 2020-08-21
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the shortcomings of current photosensitizers such as passive targeting and poor biocompatibility, and provide a water-soluble cationic porphyrin compound——tetraiodide 5,10,15,20-tetra{4-[2- (1-Methyl-1-piperidine) ethoxy] phenyl} porphyrin (hereinafter referred to as TMPiPEOPP) combined with "DNA nano-lantern" (DNA nanostructure), and applied to prepare biocompatible A nano-PDT photosensitizer that recognizes tumor cells. The nano-PDT photosensitizer has the characteristics of self-assembly, low dark toxicity to cells, high phototoxicity, no need for mandatory release of photosensitizer, and good EPR effect.

Method used

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  • Application of water-soluble cationic porphyrin in preparation of PDT nano photosensitizer
  • Application of water-soluble cationic porphyrin in preparation of PDT nano photosensitizer
  • Application of water-soluble cationic porphyrin in preparation of PDT nano photosensitizer

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Effect test

Embodiment 1

[0038] A kind of preparation of PDT nano photosensitizer specifically comprises the following steps:

[0039] (1) Four oligonucleotide sequences of A1, A2, P1, and P2 with a concentration of 1 μM were mixed with 25 mM MgCl 2 Mix with 10mM Tris-HCl buffer solution (pH 7.0) to form a mixture A; put the mixture A into a gene cycler (PCR) and heat it to 95°C for 5min, then place it on ice and incubate for 10min; Prepare DNA nano-lanterns;

[0040] (2) Dilute 2 μM P3-KRAS in 10 mM Tris-HCl buffer solution (pH 7.0) and 10 mM KCl solution to form a mixture B; put the mixture B into the gene amplification instrument and heat it to 95 ° C for 5 min, then Rapidly lower the temperature to 25°C and incubate for 0.5h; then, add 2 μM TMPipEOPP to the mixture B and mix well to obtain the composite photosensitizer TMPipEOPP / P3-KRAS;

[0041] (3) Mix the 1 μM DNA nano-lantern prepared above with the 2 μM composite photosensitizer TMPipEOPP / P3-KRAS to form a mixture C; put the mixture C into ...

Embodiment 2

[0044] The characterization of a PDT nano photosensitizer mainly adopts the following two methods:

[0045] (1) Ultraviolet-visible spectrum characterization: DNA nano-lantern, TMPipEOPP, TMPipEOPP / P3-KRAS and DNA-NPs (in which the concentration of TMPipEOPP is kept consistent) are scanned for ultraviolet-visible spectroscopy, and the obtained spectra are shown in figure 1 . It can be seen from the figure that free TMPipEOPP shows a strong Soret absorption peak at 417nm, and the centers of four weak Q absorption bands are 519, 559, 580 and 650nm respectively. When TMPipEOPP is combined with G-quadruplex P3-KRAS, a new absorption band is formed at about 700nm, and its molar absorption coefficient is much higher than that of free TMPipEOPP at 650nm, which indicates that TMPipEOPP / P3 The -KRAS complex is very promising as a composite photosensitizer, which not only redshifts the excitation light wavelength from the boundary to the interior of the biological transparent window, b...

Embodiment 3

[0048] (1) Cellular uptake and confocal microscopy imaging.

[0049] (A) Cell culture. The incubator was maintained at 37 °C, 5% CO 2 , HeLa cells were cultured in DMEM containing 10% fetal bovine serum and 1% double antibody.

[0050] (B) Adding nanocomposite photosensitizer. DNA-NPs (in which the concentration of TMPipEOPP is 0.5 μM) were co-incubated with Hela cells for 1 h, 4 h, and 8 h, respectively recorded as samples 1, 2, and 3, washed with PBS three times to remove uningested drugs, and then washed with 4 %paraformaldehyde fixed the cells for 15min.

[0051] (C) Experimental results obtained using laser confocal imaging. Olympus IX-81 microscope was used for laser confocal imaging, and 458nm was selected as the excitation wavelength of the diode-pumped laser. Obtain the confocal fluorescence microscopy images of the nanocomposite photosensitizer DNA-NPs co-incubated with HeLa cells for 1h, 4h, and 8h, see Figure 3a to Figure 3c .

[0052] (2) Detection of sing...

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Abstract

The invention discloses an application of water-soluble cationic porphyrin in preparation of a PDT nano photosensitizer. According to the method, a water-soluble cationic porphyrin compound, namely tetraiodized 5,10,15,20-tetra[1-methyl-1-piperidylethoxyphenyl]porphyrin (TMPipEOPP) 4<+>.4I<-> (hereinafter referred to as TMPipEOPP), is combined with a novel DNA nano structure; wherein the product is applied to preparation of the nano PDT photosensitizer with good biocompatibility.

Description

technical field [0001] The invention belongs to the fields of photodynamic therapy (PDT), chemical analysis and biochemistry, and particularly relates to the application of a water-soluble cationic porphyrin in preparing a PDT nano photosensitizer. Background technique [0002] Cancer is a general term for a series of related malignant tumors that can occur in almost any part of the human body. It may be related to genetic factors and long-term exposure to carcinogens. It often manifests as significant weight loss, pain, and fatigue. The specific cause of cancer is not yet clear, but as of now, there are many ways to treat cancer. Common ones are: surgery, chemotherapy, radiation therapy, photodynamic therapy and photothermodynamic therapy. Among them, photodynamic therapy (PDT) is a non-toxic drug or dye treatment that is administered systemically or locally to patients with lesions. After a period of incubation, in the presence of oxygen, red visible light with near-infra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K41/00A61K9/51A61K47/26A61P35/00C12Q1/02
CPCA61K41/0071A61K9/5123A61P35/00G01N33/5014
Inventor 朱莉娜楚俊卿孔德明王东霞
Owner TIANJIN UNIV
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