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A gene-edited microneedle for the treatment of inflammatory skin diseases and its application

An inflammatory skin disease, gene editing technology, applied in skin diseases, microneedling, gene therapy, etc., can solve the problem of difficult to reach the epidermis and dermis, and achieve the effect of improving the efficiency of gene editing and improving the inflammatory environment

Active Publication Date: 2022-06-17
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, although these small molecules have a good antagonistic effect on NLRP3, in inflammatory skin diseases, the activation of NLRP3 inflammasome is mainly located in the epidermis and dermis of human skin, and it is difficult for these inhibitors to reach the epidermis and dermis when administered orally. Dermis

Method used

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  • A gene-edited microneedle for the treatment of inflammatory skin diseases and its application
  • A gene-edited microneedle for the treatment of inflammatory skin diseases and its application
  • A gene-edited microneedle for the treatment of inflammatory skin diseases and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Preparation of Gene Editing Microneedle Patches

[0024] The present embodiment includes the following raw materials by mass percentage:

[0025] Needle tip matrix: collagen tripeptide 10%, sodium hyaluronate (molecular weight 34kDa) 10%, and the rest is water.

[0026] In addition, each 10 mL of sodium hyaluronate + collagen tripeptide solution in the microneedle tip matrix contained 60 μg of dexamethasone, 20 ug of Cas9 protein and 5 ug of sgRNA.

[0027] Base matrix: collagen tripeptide 10%, sodium hyaluronate (molecular weight 34kDa) 10%, and the rest is water.

[0028] The preparation method of the present embodiment:

[0029] 1. According to the formula ratio, disperse 1g of sodium hyaluronate (molecular weight 34kDa) and 1g of collagen tripeptide in 10mL of water, stir until fully dissolved, and add the dexamethasone nanocomposite encapsulated by the degradable polymer carrier in the prescribed proportion PLGA / Dex and a degradable cationic carrier e...

Embodiment 2

[0031] Example 2: Characterization of Gene Editing Microneedle Patches

[0032] The specific operation method is as follows: the morphology of microneedles with different percentages of collagen tripeptide was observed by scanning electron microscope. The microneedles containing different percentages of collagen tripeptide were placed on the texture analyzer, the needle tip was upward, and the probe of the texture analyzer moved downward at a speed of 1 mm / s, and the mechanical strength was reflected by measuring the curve of displacement and force.

[0033] In order to investigate the dissolution of microneedles in the skin, the double-loaded microneedle patch was applied to the back skin of mice for 0, 1, 3 and 5 min, and then removed, and then the remaining microneedle patch was taken out and photographed with a scanning electron microscope.

[0034] To evaluate the in vitro drug release behavior of the microneedles, the double-loaded microneedle patch was first immersed in...

Embodiment 3

[0036] Example 3: Drug distribution after the gene-edited microneedle patch penetrates the skin

[0037] In order to study the degradation behavior of microneedles in vivo, a double-loaded microneedle patch was prepared with PLGA / Rhodamine B and CP / Ad-SS-GD / Cas9protein-FITC, and then the double-loaded microneedle patch was applied to the back of mice At different times of the skin, the back skin tissue was isolated for cryo-embedding, and then the tissue was placed in cryosections for sectioning, and the fluorescence distribution was observed under a confocal laser microscope.

[0038] Experimental results: as figure 2 As shown, the microneedle patch can successfully penetrate the mouse skin with a depth of about 300 μm. After piercing the mouse skin, the microneedle tip part can quickly absorb the liquid in the skin tissue and dissolve quickly, while rhodamine and Cas9-FITC Large amounts are released and distributed in the epidermis and dermis. Fluorescence images at diffe...

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Abstract

The present invention provides a gene editing microneedle for treating inflammatory skin diseases and its application. The gene editing microneedle includes: a needle tip part and a base part, and the needle tip part contains two kinds of nanocomposites and a needle tip matrix. The base portion is the base matrix. The gene editing microneedles of the present invention can improve the efficiency of gene editing, deliver glucocorticoid drugs and the retention of gene editing ribonucleoprotein complexes in the skin, and synergistically improve the skin inflammation environment, and the microneedles can repair the skin barrier of skin lesions function, so as to relieve the inflammatory process of psoriasis and atopic dermatitis. The present invention also provides the application of the gene editing microneedle in preparation for treating inflammatory skin diseases.

Description

technical field [0001] The invention relates to the fields of pharmacy, biotechnology, polymer chemistry, cell biology and the like, in particular to a gene editing microneedle for treating inflammatory skin diseases and its application. It is a new type of microneedle patch to deliver gene editing ribonucleoprotein complex, and its application for preparing medicine for treating inflammatory skin diseases. Background technique [0002] Inflammatory skin diseases (ISDs) are a group of common diseases in life, and their pathological characteristics are the activation of innate and adaptive immune responses through the production of pro-inflammatory cytokines. Among various inflammatory skin diseases, psoriasis and atopic dermatitis are increasingly becoming major problems that threaten public health. Psoriasis, an immune-mediated inflammatory skin disease, is clinically characterized by marked thickening of the skin, erythematous plaques, and attached silvery scales. Psoria...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/00A61K31/573A61K48/00A61K9/51A61K47/36A61K47/42A61K47/69A61P17/06A61P17/00A61M37/00
CPCA61K9/0021A61K31/573A61K9/5153A61K47/6951A61K47/36A61K47/42A61K48/005A61P17/00A61P17/06A61M37/0015A61M2037/0053
Inventor 平渊潘齐万涛
Owner ZHEJIANG UNIV
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