Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Carrier

A carrier and delivery carrier technology, applied in the field of targeted non-viral delivery vectors, can solve the problems that basic research has not yet been reported, and siRNA delivery is at a primary level

Inactive Publication Date: 2007-12-26
IC VEC LTD
View PDF18 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although the delivery of many different nucleic acids has been extensively described, research into siRNA delivery is still at a rudimentary level
Thus, despite widespread use of cationic lipid / liposome systems for the delivery of plasmid DNA (pDNA) and oligodeoxynucleotides (ODN) to cells (7, 9, 25-28), there is little in the literature about lipids containing cationic siRNA formulation of lipid / liposome and its delivery to cells in vitro or in vivo (siFection)
Even, fundamental studies involving the formulation of cationic lipid / liposome systems containing siRNA have yet to be reported

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
  • Carrier
  • Carrier
  • Carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0449] PEGylated siRNA-lipoplex

[0450] Referring to accompanying drawing IA, DOPE (140 μ L, in CHCl 3 10.68mg / mL in CHCl), CDAN·3HCl (271μL, in CHCl 3 4mg / mL) and CPA (100μL, in CHCl 3 4.4 mg / mL) in a round-bottomed flask (5 mL) pretreated with nitric acid and dimethylsilyl dichloride, and the solvent was removed under reduced pressure to form a lipid film by adding water under vortexing (milliQ, 1 mL) to rehydrate. Then in the Sonomatic TM Unilamellar liposomes (SUV) were obtained by sonicating the multilamellar liposome formulation for 30 minutes in a water bath (Longford Ultrasonics). 250 μL of these liposomes were pipetted into 5 mL Falcon plastic tubes and siRNA solution (0.28 mg / mL) was added dropwise under vigorous vortexing, followed by polyethylene glycol-bisaldehyde (Mw 3400, 7.8 μL, 10 mg / mL; 5% PEG / total lipid). Samples were kept stable for 15 min / RT after which time PBS (483 μL) was added. After keeping the samples for 16 hours / RT, the volume was reduced ...

Embodiment 2

[0453] Stability of PEGylated siRNA-lipoplex.

[0454] The stability of PEGylated (Mw 2000) siRNA-lipoplex in 80% serum (FCS) was studied. Surface PEGylated LsiR complexes were generated as described in Example 1 and Figure 1 and incubated with FCS for various times, after which particle size was determined by photon correlation spectroscopy (PCS). The results are shown in Figure 2. As the degree of PEG increases, particle size does not increase over the timescale studied.

[0455] Advantageously, PEGylated siRNA-lipoplexes produced by coupling PEG to siRNA-loaded lipoplexes exhibit serum stability with increasing amounts of PEG coupled to the surface.

Embodiment 3

[0457] Tissue distribution studies

[0458] Using lipids [4- 14 C] Cholesterol (Amersham Biosciences) CDAN / DOPE / CPA700 / [4- 14 C] Cholesterol-labeled liposome formulation. siRNA lipoplexes were prepared by adding siRNA to liposomes at a liposome / siRNA ratio of 13:1 (w / w) under vortexing. Samples were concentrated to half of the total volume and replenished to the original volume by adding PBS. 200 μL (0.1 mg / mL siRNA) of these complexes were injected into the caudal vein of each mouse weighing approximately 30 g. Radioactivity was adjusted to approximately 0.035 μCI / animal.

[0459] After 1 hour, mice were anesthetized and blood was obtained by cardiac puncture and immediately mixed with 15 U heparin. Blood concentrations of liposomes were calculated assuming a total blood weight of 6% of body weight. After cervical dislocation, liver, spleen, kidney, lung and heart were dissected and weighed. Homogenize the organs in PBS at a concentration of 5 mL PBS / g organ. Aliquots...

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

PropertyMeasurementUnit
diameteraaaaaaaaaa
Login to View More

Abstract

The present invention relates to a non-viral delivery vector comprising a liposome, wherein one or more lipids of the liposome are coupled, reversibly or irreversibly, to one or more polymers, and wherein the liposome comprises siRNA.

Description

technical field [0001] The present invention relates to non-viral delivery vehicles comprising siRNA. The invention also relates to targeted non-viral delivery vectors, methods of making these vectors, methods of using these vectors, and uses thereof. Background technique [0002] There are many problems with viral DNA delivery methods, including immune response, inability to repeatedly deliver viral DNA vectors, difficulty in generating high viral titers, and potential for viral infection. Non-viral delivery methods provide an alternative system that does not have these problems and thus facilitates the development of low-risk, non-viral means for gene transfer. [0003] Nonviral delivery systems with great potential include the use of cationic liposomes, which are usually composed of neutral phospholipids and cationic lipids. They have been used to deliver DNA, mRNA, antisense oligonucleotides, proteins and drugs into cells. Many cationic liposomes are commercially avai...

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
Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K9/127C12N15/88
CPCA61K48/0091Y10T428/2984A61K48/00C12N15/88A61K9/1272A61K2121/00A61P43/00
Inventor M·凯勒M·乔根森A·D·米勒E·佩罗泽尔
Owner IC VEC LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com