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Targeted drug delivery system modified by follicle-stimulating hormone polypeptide and regulated by promoter MUC16 and preparation method of targeted drug delivery system

A follicle-stimulating hormone and drug delivery system technology, which is applied in the field of in vivo and in vitro curative effect investigation of the targeting system, can solve the problems of recurrence or drug resistance, treatment failure and other problems of advanced patients, and achieve reduction of non-specific uptake, minimization of toxic and side effects, The effect of increasing specific uptake

Inactive Publication Date: 2017-01-18
THE OBSTETRICS & GYNECOLOGY HOSPITAL OF FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Most patients with ovarian cancer are diagnosed at an advanced stage. Chemotherapy is an important means of treatment for advanced ovarian cancer. However, most patients with advanced ovarian cancer relapse or become drug-resistant after treatment, resulting in treatment failure.

Method used

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  • Targeted drug delivery system modified by follicle-stimulating hormone polypeptide and regulated by promoter MUC16 and preparation method of targeted drug delivery system
  • Targeted drug delivery system modified by follicle-stimulating hormone polypeptide and regulated by promoter MUC16 and preparation method of targeted drug delivery system
  • Targeted drug delivery system modified by follicle-stimulating hormone polypeptide and regulated by promoter MUC16 and preparation method of targeted drug delivery system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] targeted detection

[0047] Ovarian cancer cells HEY and SKOV3 were cultured in vitro, and nanocomplexes regulated by FSH polypeptide modification and / or MUC16 promoter (FSH33- Silencing effect of G-NP, FSH33-MUC16.1 / G-NP and FSH33-MUC16.2 / G-NP) on the target gene gro-α ( image 3 , Figure 4 ). The results showed that in FSHR-positive MUC16-positive HEY cells, the down-regulation effect of FSH33-MUC16.1 / G-NP and FSH-MUC16.2 / G-NP on gro-α mRNA was higher than that of other groups, and the gro in the supernatant The level of -α was significantly lower than that of other groups, and the difference was statistically significant. After 48 hours of transfection, FSH33-MUC16.1 / G-NP can significantly reduce the level of gro-α protein in the cytoplasm. In FSHR-negative MUC16-positive SKOV3 cells, after FSH33-MUC16.1 / G-NP transfection for the same time, the down-regulation effect of gro-α mRNA, the level of gro-α in supernatant and the level of gro in cytoplasmic protein amo...

Embodiment 2

[0049] Evaluation of the Effects of Nanocomplexes on the Malignant Biological Behavior of Ovarian Cancer Cells

[0050] The specific steps of the cytotoxicity test were operated according to the instructions of the CCK8 kit. Figure 5 The proliferation activity of HEY and SKOV3 cells was shown at 24h, 48h, 72h and 96h after treatment with different groups of nanocomposites. In FSHR-positive and MUC16-positive ovarian cancer HEY cells, the proliferative activity of cells was differently inhibited after FSH33-MUC16.1 / G-NP, FSH33-MUC16.2 / G-NP and FSH33-G-NP nanocomplexes. Among them, the inhibitory effect of FSH33-MUC16.1 / G-NP is the most obvious, and the cell activity after 96 hours of treatment is only 25.32% of that of the control group, and the difference is statistically significant compared with other groups. In the FSHR-negative and MUC16-positive ovarian cancer SKOV3 cells, the cell viability was not significantly inhibited after transfection with the drugs of each group...

Embodiment 3

[0053] Preliminary evaluation of in vivo pharmacodynamics and safety of targeted nanocomplexes

[0054] Replicate human ovarian cancer subcutaneous xenograft tumor model, divided into 6 groups: targeting complex modified by FSH polypeptide and regulated by MUC16.1 promoter (5% FSH33-MUC16.1 / G-NP); nanocomposite modified by FSH polypeptide Object group (5%FSH33-G-NP), nanocomplex barrier without FSH polypeptide modification (MUC16.1 / G-NP); naked plasmid group (pDNA); blank nanoparticle group (NP), solvent control group ( Control).

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Abstract

The invention provides a targeted drug delivery system modified by a follicle-stimulating hormone polypeptide and regulated by a promoter MUC16 and a preparation method of the targeted drug delivery system. The system is a two-way target system, comprises a molecular target, a gene target, a nano-carrier and a small interfering RNA, and relates to the follicle-stimulating hormone (FSH), a mucoprotein antigen MUC16 and a growth regulatory cancer gene Gro-alpha. The preparation method of the system comprises the following steps: (1) preparing the two-way targeted drug delivery system from follicle-stimulating hormone polypeptide, the mucoprotein antigen MUC16, alpha-succinimidyl-omega-propyl maleimide groups-polyethylene glycol and polyethylene imine branches, and polyethylene glycol (PEG); (2) introducing the small interfering RNA of the gro-alpha into the drug delivery system to serve as a model drug. The advantages are as follows: the targeted drug delivery system modified by the follicle-stimulating hormone polypeptide is constructed, a nanomaterial is used as a drug carrier, the gro-alpha shRNA is an ammunition, the distribution of the drug in an FSHR positive ovarian cancer cell is improved, and the effect of gro-alpha secreted by the ovarian cancer cell on a ground substance is blocked, so that the growth and propagation of a tumor are suppressed.

Description

technical field [0001] The present invention relates to a method for the expansion and preparation of a targeted drug delivery system modified by follicle-stimulating hormone polypeptide and regulated by the MUC16 promoter, especially related to the in vivo and in vitro curative effect of the targeted system using gro-α shRNA as a model drug study. Background technique [0002] The mortality rate of ovarian cancer ranks among the top five among women's cancer deaths, and ranks first among gynecological malignancies. Most patients with ovarian cancer are diagnosed at an advanced stage. Chemotherapy is an important means of treatment for advanced ovarian cancer. However, most patients with advanced ovarian cancer relapse or become drug-resistant after treatment, leading to treatment failure. Therefore, it is necessary to explore new treatment methods for ovarian cancer. Targeted therapy has become a hot spot in ovarian cancer treatment research. [0003] Targeted therapy ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/42A61K47/34A61K47/10A61K31/7105A61K47/62A61K48/00A61P35/00
CPCA61K47/42A61K31/7105A61K47/10A61K47/34
Inventor 徐丛剑张晓燕张明星洪珊珊张梦宇张萌
Owner THE OBSTETRICS & GYNECOLOGY HOSPITAL OF FUDAN UNIV
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