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A kind of photosensitizer compound and its preparation method and application

A technology for photosensitizers and complexes, applied in the field of photosensitizer complexes and their preparation, can solve the problems of being unsuitable for the treatment of hematological tumors, losing the selectivity of the illumination area, etc., and achieves enhanced stability, improved targeting, and high safety. Effect

Active Publication Date: 2020-06-30
李斯文 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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

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  • A kind of photosensitizer compound and its preparation method and application
  • A kind of photosensitizer compound and its preparation method and application
  • A kind of photosensitizer compound and its preparation method and application

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preparation example Construction

[0048] The present invention also provides a method for preparing the photosensitizer complex according to the above technical solution, which includes the following steps:

[0049] 1) Mixing chitosan and hydrophobic compounds in a mass ratio of (20-30):4 for esterification reaction to obtain a grafted polymer, and removing residual hydrophobic compounds by dialysis;

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

[0051] 3) The photosensitizer-loaded micelles described in step 2) are subjected to an esterification reaction with a specific antibody to obtain a photosensitizer complex.

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

Embodiment 1

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

[0076] Obtain the VH sequence (shown in SEQ ID NO: 1) and VL sequence (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 and N-terminus of VH to obtain the sequence of anti-CD19 single-chain antibody (MEDI-551-scFv) (shown in SEQ ID NO: 3), and add BamHI and HinderIII to the 2 ends of the sequence The restriction site was inserted into the expression vector pET-SUMO in the manner of enzyme digestion. After colony PCR verification and sequencing, the correct recombinant expression vector was screened: 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), and the correct expres...

Embodiment 2

[0079] Obtaining of MEDI-551-scFv protein:

[0080] The expression strain (Origami 2(DE3) / pET-SUMO-MEDI-551-scFv) was subjected to a 2L large-scale fermentation to extract its cytoplasmic protein. Since the target protein obtained by the expression vector has a his tag at the 5'end, And there is a section of SUMO restriction site afterwards, so the corresponding single-chain antibody (MEDI-551-scFv) protein was obtained by Ni column purification-restriction 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 pellet.

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

[0084] 3) After the above mixture was magnetically stirred at 4°C for 30 minutes, centrifuged at 14000 rpm for 30 minutes, and the supernatant was collected and filtered wit...

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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 complex and a preparation method and application thereof. 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 the absorption of photons to the surrounding oxygen atoms to oxidize it 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 such photosensitizers can significantly extend the circulation time of photosensitizers in the body, reduce the amount of photosensitizers themselves, and increase the proportion of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/36A61K41/00A61K47/69A61K47/68A61P35/02
CPCA61K41/0071
Inventor 李斯文赵磊
Owner 李斯文
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