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Application of chitosan nanoscale ultrasound contrast agent combined with dickkopf-2 gene in the preparation of drugs for the treatment of prostate cancer

A technology of chitosan nano and ultrasound contrast agent, which is applied in the field of biomedicine, can solve the problems of high mortality and poor treatment effect, and achieve the effect of good biocompatibility, low cost and favorable targeted delivery

Active Publication Date: 2020-09-04
SHANDONG UNIV QILU HOSPITAL
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Surgical treatment of patients with early prostate cancer has a good prognosis, and castration-based comprehensive endocrine therapy is often used in the middle and late stages. Most patients will gradually transform into castration-resistant prostate cancer (CRPC), with a high mortality rate and curative effect. Difference

Method used

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  • Application of chitosan nanoscale ultrasound contrast agent combined with dickkopf-2 gene in the preparation of drugs for the treatment of prostate cancer
  • Application of chitosan nanoscale ultrasound contrast agent combined with dickkopf-2 gene in the preparation of drugs for the treatment of prostate cancer
  • Application of chitosan nanoscale ultrasound contrast agent combined with dickkopf-2 gene in the preparation of drugs for the treatment of prostate cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1: the preparation of chitosan nanoscale ultrasonic contrast agent

[0052] The preparation of chitosan nanoscale ultrasound contrast agent, the steps are as follows:

[0053] (1) 50 μ L of acetic acid is added to 5 mL of deionized water, mixed upside down, the preparation volume fraction is 1% acetic acid solution, chitosan is added in the acetic acid solution, chitosan solution is prepared after dissolving chitosan, The concentration of chitosan in the chitosan solution is 2g / L;

[0054] (2) Disperse 0.15mL perfluorohexane, 0.006mL Tween 20 and 0.004g lecithin in 2.7mL deionized water to make a suspension, and homogenize the suspension in a FJ2000-S high-speed dispersion homogenizer After 1 min, the rotation speed was 19000r / min, and the emulsified product Ⅰ was obtained;

[0055] (3) Add the chitosan solution prepared in step (1) dropwise to the emulsified product I prepared in step (2), and homogenize in a high-speed dispersing homogenizer for 2 minutes...

Embodiment 2

[0058] Embodiment 2: the characterization of chitosan nanoscale ultrasonic contrast agent (CND)

[0059] 1. Observation of surface morphology and composition by optical microscope, transmission electron microscope and scanning electron microscope

[0060] Observe after diluting the CND prepared by embodiment 1 with physiological saline, the optical microscope photo of CND is as follows: figure 2 As shown, under a 1000× optical microscope, CND is circular, with uniform particle size, uniform dispersion and no aggregation; the transmission electron microscope photos of CND are as follows image 3 As shown, the CND is round under the transmission electron microscope, and the surface is smooth and bright; the scanning electron microscope photo of the CND is as follows Figure 4 As shown, the CND presents a regular spherical shape under the scanning electron microscope.

[0061] 2. Particle size and potential

[0062] After diluting the CND prepared in Example 1 with pure water...

Embodiment 3

[0063] Embodiment 3: Gene binding and protection ability evaluation of chitosan nanoscale ultrasound contrast agent (CND)

[0064] CND and plasmid DNA (pDNA) prepared in Example 1 were mixed and incubated at room temperature for 30 minutes according to different mass ratios to prepare CND / pDNA complexes, wherein the mass ratios of pDNA and CND were 1:1, 1:5, and 1:10, respectively. , 1:20, the quality of pDNA was 1 μg.

[0065] The particle size and Zeta potential of the above CND / pDNA complex were detected, and the results are shown in Table 1.

[0066] Table 1. Particle size and Zeta potential of CND / pDNA complexes with different mass ratios

[0067]

[0068] In the prepared CND / pDNA complex, the larger the pDNA addition ratio, the larger the particle size of the CND / pDNA complex, indicating that the more pDNA is added, the more pDNA is electrostatically bound to the surface of CND; in addition, with the addition of pDNA As the amount increases, the surface potential of...

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Abstract

The invention relates to an application of united use of a chitosan nanometer-level acoustic contrast agent and Dickkopf-2 gene to preparation of medicines for treating prostate cancer. For the firsttime, the inventor finds relationship of Dickkopf-2 gene and prostatic cancer, the expression of the Dickkopf-2 gene in tissues of patients suffering from prostatic cancer is obviously reduced, the chitosan nanometer-level acoustic contrast agent is used as a transmission carrier, the Dickkopf-2 gene is directionally conveyed to tumor tissue, and through expression of the Dickkopf-2 gene, proliferation of tumor cells is restrained. The prepared chitosan nanometer-level acoustic contrast agent adopts chitosan as shell membrane materials, and is safe, non-toxic, good in biocompatibility and lowin cost. The particle diameter is in the nanometer-level scope, and the chitosan nanometer-level acoustic contrast agent can smoothly pass through the gap of vascular wall of tumor tissue, so as to deliver target genes to the tumor tissue, and through united use of an ultrasonic target destroy microvesicle technique, the gene transfection efficiency can be notably improved.

Description

technical field [0001] The invention relates to the application of chitosan nanoscale ultrasonic contrast agent combined with Dickkopf-2 gene in the preparation of drugs for treating prostate cancer, and belongs to the technical field of biomedicine. Background technique [0002] With the continuous breakthrough of key technologies of gene therapy, there will be more new options for clinical treatment of various diseases. Many diseases have well-defined lesion sites, especially tumors. Therefore, it is very important to find a method that can effectively deliver the therapeutic gene to the lesion site. Over the past few decades, viral vectors have dominated the field of gene therapy due to their high gene delivery efficiency. However, its security issues limit its applications. This limitation of viral vectors has accelerated the search for alternative nonviral delivery vectors. Among non-viral vectors, ultrasound contrast agents are a promising gene delivery system that...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K47/69A61P35/00A61K49/22
CPCA61K47/6939A61K48/0025A61K48/005A61K49/221A61K49/225A61P35/00
Inventor 李杰刘欣欣时丹丹郭鲁周晓莹
Owner SHANDONG UNIV QILU HOSPITAL
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