Composition comprising endostatin adopted as delivery system and chemically-synthesized RNA interference molecule, and application thereof

A technology of RNA interference and endostatin, which is applied in the field of compositions composed of RNA interference molecules, can solve the problems of high toxicity and side effects, high manufacturing cost, poor stability, etc., and achieve the effect of small toxicity and side effects, good therapeutic effect and stable activity

Active Publication Date: 2012-11-21
NANJING UNIV +1
View PDF4 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The purpose of the present invention is to solve the problems of high dosage, poor stability, relatively large toxic and side effects or high manufacturing cost when siRNA, asiRNA, and miRNA gene drugs characteriz

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Composition comprising endostatin adopted as delivery system and chemically-synthesized RNA interference molecule, and application thereof
  • Composition comprising endostatin adopted as delivery system and chemically-synthesized RNA interference molecule, and application thereof
  • Composition comprising endostatin adopted as delivery system and chemically-synthesized RNA interference molecule, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Schematic diagram of the process of combining endostatin and siRNA in the presence of liposomes through positive and negative charges to form a complex ( figure 1 ) said that the advantage of dividing siRNA, endostatin, and liposome into three bottles is that the quality control of each component is easier and the storage is more convenient, but it is more troublesome to use. The ingredients of the three bottles can also be pre-mixed together according to a certain charge ratio described in the following examples, and combined into one bottle, which is convenient to use, but the quality control of the mixed components is more difficult. figure 1 Shown is the siRNA of tumor anti-apoptosis gene (B-cell lymphoma / le μkemia-2, Bcl2, formerly known as B-cell lymphoma / leukemia 2 gene, Bcl2), which can be replaced by asiRNA or miRNA of this gene or other genes , figure 1 The exemplified liposomes may be cationic liposomes, neutral liposomes, or PEG-modified long-acting liposom...

Embodiment 2

[0066] The endostatin recombinant protein used in this example is produced by Zhongkai Biopharmaceutical Factory of Jiangsu Wuzhong Pharmaceutical Group Co., Ltd., hereinafter referred to as endostatin, which consists of 184 amino acids. The amino acid sequence is: SEQ ID NO.1, isoelectric The point is 9.3. At pH5.0, the charge of endostatin is +12.89; at pH5.5, the charge of endostatin is +10.84; at pH6, the charge of endostatin is +8.65 ( figure 2 ). Use endostatin with pH 5.5 and charge value of +10.84, and Bcl2-siRNA with charge value of -40 (sequence: sense strand 5'CGGAGGCUGGGAUGCCUUUUdTdT 3', antisense strand 3'dTdTGCCUCCGACCCUACGGAAA 5'), through positive and negative Charge calculation binding ratio, the mixture of Bcl2-siRNA, endostatin, PEG liposome (polyethylene glycol 2000-distearoylphosphatidylethanolamine (PEG-PE) and dioleylphosphatidylserine (DOPS), The mass ratio of PEG-PE to DOPS is 1:1, and the final concentration is 1mg / ml, the same below) are mixed in ...

Embodiment 3

[0069] In addition to detecting the zeta potential in the case of liposome encapsulation according to the method of Example 2, to determine the charge value of the composition of the endostatin recombinant protein and the RNA interference molecule after the combination of positive and negative charges, it can also be obtained by electrophoresis Methods To determine the charge of the composition composed of endostatin recombinant protein and RNA interference molecules with or without liposome encapsulation. Prepare TBE solution, Tris 10.8g, boric acid 5.5g, EDTA 0.58g, add sterilized DEPC water to make up to 1000ml. Weigh 0.6g of agarose, dissolve it in 60ml of TBE solution, heat for two minutes, add a small amount of EB after cooling, pour it into the gel-making tank, and insert a comb to make an agarose gel. Gently mix 1.5 μl of Bcl2-siRNA (sequence and charge value are the same as in Example 2, and the concentration is 20nM) with endostatin (sequence and charge value are the...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The present invention belongs to the field of biomedicine, and discloses a composition comprising endostatin adopted as a delivery system and a chemically-synthesized RNA interference molecule, and an application thereof. The composition contains endostatin recombination protein carrying a charge of +5 to +15, and a chemically-synthesized RNA interference molecule carrying a charge of -15 to -55, wherein the endostatin recombination protein and the chemically-synthesized RNA interference molecule are subjected to positive charge and negative charge combination to form a composition with a charge value ??of -20 to +20. With the composition, use amounts and charge values of the endostatin and the chemically-synthesized RNA interference molecule are greatly reduced; the endostatin providing a delivery effect has a special characteristic of lesion vascular endothelial cell targeting, such that the composition can target the target tissue and stably distribute after the composition is subjected to intravenous injection 1 hour or within 24 hours so as to provide advantages of good treatment effect, stable activity, long effective action time and low toxic and side-effect.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a composition composed of endostatin as a delivery system and a chemically synthesized RNA interference molecule and its application. Background technique [0002] Endostatin (endostatin) was first produced by primary tumor cells and can inhibit the growth of tumor blood vessels (O'Reily et al. 1997, Cell 88:277). It is a fragment of collagen XVIII, composed of 183 amino acids, with a high positive Charge is a highly positively charged protein that is rarely seen in recombinant proteins. At present, 183 amino acids, 184 amino acids, 193 amino acids with 6 histidines at the N-terminal and PEG-modified endostatin have been clinically studied or have been marketed. However, there is no corresponding report on endostatin as a delivery system to deliver chemically synthesized RNA interference molecules to corresponding target tissues. [0003] It is currently known that endostatin has the f...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): A61K48/00A61K31/713A61K47/48A61P1/16A61P31/14A61P31/20A61P35/00A61P31/18
Inventor 徐根兴殷妍郭佳佳张昌栋吴稚伟
Owner NANJING UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap