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SiRNA and anticancer drug targeting co-delivery system as well as preparation method and application thereof

A co-delivery and anti-cancer drug technology, applied in the field of siRNA and anti-cancer drug targeted co-delivery system and its preparation, to achieve the effects of improving gene transfection efficiency, strong cell membrane adhesion, and reducing cytotoxicity

Active Publication Date: 2021-08-13
THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the combination of chemotherapy and gene therapy faces a huge challenge, that is, the synthesis of safe, efficient and specific nanocarriers

Method used

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  • SiRNA and anticancer drug targeting co-delivery system as well as preparation method and application thereof
  • SiRNA and anticancer drug targeting co-delivery system as well as preparation method and application thereof
  • SiRNA and anticancer drug targeting co-delivery system as well as preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] The present invention will be further described below in conjunction with specific embodiments, but the protection scope of the present invention is not limited to the following embodiments.

[0060] The materials used in the following examples are as follows

[0061] Polyethyleneimine (PEI) with an average molecular weight of 1800 and 25000 was purchased from Aldrich Company. Sodium alginate, viscosity 300 cps (Lot M3H5540, Nacalaitespue INC., Kyoto, Japan). Cholesterol Chloroformate Alfa Aesar. Dichloromethane was purchased from Guangzhou Reagent Co., Ltd. Oxidized sodium alginate, self-made in the laboratory. Adriamycin hydrochloride was purchased from Guangzhou Eskin Biotechnology Co., Ltd. siRNA was purchased from Guangzhou Ruibo Biotechnology Co., Ltd. iRGD was purchased from Jill Biochemical (Shanghai) Co., Ltd. RIPA, purchased from CST Corporation. MCF-7 was purchased from the Cell Bank of the Chinese Academy of Sciences. Resazurin was purchased from Gua...

Embodiment 2

[0132] On the surface of tumor cells and neovascular endothelial cells, the most widely and highly expressed integrin is αVβ3, especially in the process of tumor angiogenesis, αVβ3 plays an important role. The αVβ3 receptor is generally not expressed or less expressed in mature vascular endothelial cells and most normal tissues, but it is highly expressed on the surface of neovascular endothelial cells in tumor tissues and various malignant tumor cells.

[0133] Neuropilin-1 (NRP-1) is a transmembrane glycoprotein located on the forming nerve fiber axon, NRP-1 is highly expressed in a variety of tumors, such as pancreatic cancer, lung cancer, Prostate cancer, ovarian cancer and stomach cancer etc. The survival and growth of various tumor cells are heavily dependent on the expression of the cell's own NRP-1. This application uses Western Blot technology to screen tumor cells that highly express αVβ3 receptors and NRP-1, and then explores the ability of DOX / siRNA / iRGD-NPs to ta...

Embodiment 3

[0169] Analysis of targeted penetration ability of DOX / siRNA / iRGD-NPs tumorspheres

[0170] 1. Tumorsphere culture

[0171] In this example, MCF-7 was selected as the target cell, and the tumor sphere cell model was constructed by combining the hanging drop method and the suspension culture method. FAM-siRNA was used as the fluorescent indicator to explore the targeting of tumor spheres in vitro by NPs modified by iRGD. Penetrating ability.

[0172] The tumor spheres were cultured using conventional methods.

[0173] 2. Tumorsphere penetration test

[0174] Using FAM-siRNA as a fluorescent indicator, the tumor spheres and the complex solution FAM-siRNA-NPs, FAM-siRNA / iRGD-NPs in 5% CO 2 After incubating in a 37°C incubator for 4 hours, gently wash the tumor spheres with pre-cooled PBS 3 times, then add 200 μL of 4% paraformaldehyde to fix for 30 minutes, then use PBS to gently wash 3 times, and place them in PBS solution for 4 hours. Store at ℃ for subsequent laser confoca...

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Abstract

The invention discloses a siRNA and anticancer drug targeted co-delivery system and a preparation method thereof. The tumor targeted cell-penetrating peptide iRGD is adopted for conducting targeted modification on NPs, then chemotherapy drug doxorubicin hydrochloride and siRNA of vascular endothelial growth factors are loaded, a tumor treatment system DOX / siVEGF / iRGD-NPs is constructed, the system has relatively low cytotoxicity; and through the synergistic effect of DOX, siVEGF, iRGD and NPs, the ability of active targeting inhibition of tumor cells MCF-7, neovascularization and targeting penetration of tumor spheres is obtained, and the potential application prospect in targeting therapy of tumors is predicted.

Description

technical field [0001] The invention belongs to the technical field of targeted inhibition of tumor cells, and specifically relates to a targeted co-delivery system of siRNA and anticancer drugs, a preparation method and application thereof. Background technique [0002] Tumor has become one of the three major diseases in the world due to its high morbidity and mortality, which seriously threatens human life and health. In recent years, combined chemotherapy and gene therapy for tumor treatment has become a research hotspot at home and abroad. Chemotherapy, as one of the most important methods of tumor treatment, can be used to treat various types of tumors, but because drugs within a safe dose range cannot completely eradicate tumors, excessive non-specific and selective drugs accumulate in the body, causing severe toxicity. side effect. The addition of therapeutic genes can reduce the dosage of chemotherapeutic drugs, thereby reducing the toxic and side effects caused by...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/127A61K47/36A61K31/704A61K47/64A61K48/00A61P35/00C08G81/00
CPCA61K31/704A61K9/1273A61K47/64A61K48/005A61K48/0041A61P35/00C08G81/00Y02A50/30
Inventor 王琴梅
Owner THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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