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Photosensitizer compound and preparation method and application of photosensitizer compound

A photosensitizer and compound technology, applied in the field of photosensitizer compound and its preparation, can solve the problems of being unsuitable for the treatment of blood tumors and losing the selectivity of the light area, achieving enhanced stability, improved targeting, and high safety Effect

Active Publication Date: 2017-09-01
李斯文 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because lymphoma cells are diffusely distributed in the blood, PDT treatment of hematoma requires irradiation of the blood, which loses the selectivity of the light area, which makes the above-mentioned photosensitizer with chitosan as the carrier, even if it has targeting and Good controlled release, not suitable for the treatment of hematoma

Method used

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  • Photosensitizer compound and preparation method and application of photosensitizer compound
  • Photosensitizer compound and preparation method and application of photosensitizer compound
  • Photosensitizer compound and preparation method and application of photosensitizer compound

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preparation example Construction

[0048] The present invention also provides a preparation method for the photosensitizer complex described in the above technical solution, comprising the following steps:

[0049] 1) Chitosan and hydrophobic compound are mixed according to mass ratio (20~30): 4 to carry out esterification reaction, obtain graft polymer, remove residual hydrophobic compound by dialysis;

[0050] 2) Dissolving the graft polymer described in step 1) in a polar solution to obtain a graft polymer solution with a concentration of 0.5-1.5 mg / mL, adding the photosensitizer dropwise to the graft polymer at a rate of 20-40 μL / min In the solution, micelles loaded with photosensitizer are obtained;

[0051] 3) Esterifying the photosensitizer-loaded micelles described in step 2) with the specific antibody to obtain a photosensitizer complex.

[0052] In the invention, the chitosan and the hydrophobic compound are mixed according to the mass ratio of (20-30): 4 to carry out the esterification reaction to o...

Embodiment 1

[0075] Construction of MEDI-551-scFv expression vector:

[0076] Obtain the VH sequence (as shown in SEQ ID NO:1) and VL sequence (as shown in SEQ ID NO:2) of the anti-CD19 antibody (MEDI-551) through drugbank, and then use the overlap PCR method to pass a flexible peptide ( G 4 S) 3 Connect the C-terminus of VL to the N-terminus of VH to obtain the sequence of the anti-CD19 single-chain antibody (MEDI-551-scFv) (as shown in SEQ ID NO: 3), and add BamHI and HinderIII to the 2 ends of the sequence Restriction sites were inserted into the expression vector pET-SUMO by restriction enzyme-linking. After colony PCR verification and sequencing, the correct recombinant expression vector was screened out: pET-SUMO-MEDI-551-scFv electrophoresis results are as follows: figure 1 As shown, Lane1 is a single-chain antibody (MEDI-551-scFv); Lane2 is a recombinant expression vector (pET-SUMO-MEDI-551-scFv).

[0077] The recombinant expression vector was transformed into Origami 2(DE3), an...

Embodiment 2

[0079] Obtaining MEDI-551-scFv protein:

[0080] The expression strain (Origami 2(DE3) / pET-SUMO-MEDI-551-scFv) was subjected to 2L large-scale fermentation, and its cytoplasmic protein was extracted. Since the 5' end of the target protein obtained by the expression vector has a his tag, And thereafter contains a section of SUMO enzyme cleavage site, so the corresponding single-chain antibody (MEDI-551-scFv) protein was obtained by Ni column purification-enzyme digestion-Ni column repurification.

[0081] Specific steps are as follows:

[0082] 1) Collect the cultured Escherichia coli and centrifuge at 8000 rpm for 30 min at 4°C to obtain bacterial pellets.

[0083] 2) Resuspend the pellet in 20 mL of cell lysate (50 mM Tris-HCl, 0.5 M NaCl, 1% TritonX-100, pH 8.0) containing 0.1 mg / mL lysozyme, and sonicate for 10 min.

[0084] 3) The above mixture was magnetically stirred at 4° C. for 30 minutes, then centrifuged at 14,000 rpm for 30 minutes, and the supernatant was collect...

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Abstract

The invention relates to a photosensitizer compound, and a preparation method and an application of the photosensitizer compound and belongs to the technical field of photodynamic treatment. The photosensitizer compound is in a shell core structure and takes chitosan as a hydrophilic framework; a hydrophobic compound is coupled into the chitosan hydrophilic framework via an amido bond; the hydrophobic compound is coupled with a photosensitizer to form a core structure; and the exterior of the chitosan hydrophilic framework has an action with a polar water molecule and is further modified by a specific antibody to form a shell structure. The photosensitizer compound is good in targeting ability and high in safety, and can allow selectivity of the photodynamic treatment not to depend on an illumination area, and the photodynamic treatment to have cell selectivity; only the photosensitizer uptaken by cells can generate photoinduced toxicity; and the photosensitizer be well applied to treatment of hematologic malignancies and also has a very wide application prospect in preparation of acute B lymphocyte leukemia drugs.

Description

technical field [0001] The invention belongs to the technical field of photodynamic therapy, and in particular relates to a photosensitizer compound and its preparation method and application. Background technique [0002] The principle of photodynamic therapy (PDT) is that after a photosensitizer absorbs light of a specific wavelength, it can transfer the ability generated by absorbing photons to the surrounding oxygen atoms, oxidizing them into active oxygen, which then oxidizes lipids and proteins. , Nucleic acid biomolecules can kill cells. Oxygen, photosensitizer, and light of a certain wavelength are the three major elements of PDT treatment. [0003] The existing photosensitizer complex prepared by using chitosan as a carrier is a photosensitizer used in photodynamic therapy. Although this type of photosensitizer can significantly prolong the circulation time of the photosensitizer in the body, reduce the amount of the photosensitizer itself, and increase the propor...

Claims

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Application Information

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IPC IPC(8): A61K41/00A61K47/69A61K47/68A61P35/02
CPCA61K41/0071
Inventor 李斯文赵磊
Owner 李斯文
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