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

Positive ion nanostructure lipid carrier, manufacturing method and application thereof

A cationic lipid and carrier technology, applied in the field of biomedicine, can solve the problems of incomplete siRNA loading, and achieve the effects of low toxicity, inhibition of expression, and high transfection efficiency

Active Publication Date: 2013-02-13
BEIJING INTELLIGENE BIOTECH LTD CO
View PDF2 Cites 12 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

3. Cationic lipid carriers often have incomplete loading of siRNA, resulting in siRNA being degraded by serum proteins

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
  • Positive ion nanostructure lipid carrier, manufacturing method and application thereof
  • Positive ion nanostructure lipid carrier, manufacturing method and application thereof
  • Positive ion nanostructure lipid carrier, manufacturing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] The synthesis of embodiment 1 compound VI

[0085] All solvents, raw materials and reagents are analytically or chemically pure unless otherwise specified. The anhydrous treatment of the solvent is carried out according to a conventional method.

[0086] A. Using the structure of formula I with Boc 2 O is reacted to give formula II.

[0087]

[0088]

[0089] B. Formula II is reacted under the conditions of DCC and DMAP to obtain formula III with a hydrophobic tail.

[0090]

[0091] C. Formula III removes the Boc protecting group under the condition of TFA to obtain formula IV.

[0092]

[0093] D. Formula IV reacts under certain conditions to obtain formula V with a hydrophilic head.

[0094]

[0095] E. Under the condition of TFA, formula V removes the Boc protecting group to obtain formula VI.

[0096]

[0097] F. The synthetic route, reagents and conditions are as follows:

[0098]

[0099] Reagents and conditions: a.Boc 2 O,NaOH,H 2 O,...

Embodiment 2

[0100] Embodiment 2 is used for the preparation of the liposome of nucleic acid transfection

[0101] Weigh 4.2 mg of compound VI in a silanized Erlenmeyer flask, dissolve in 1 mL of organic solvent (chloroform / methanol=1:1, v / v), shake and dissolve, and dissolve at a flow rate of 5 kg / cm 2 Blow dry the solvent with nitrogen or argon gas to make the dissolved matter into a thin film. The residual solvent is slowly evaporated under reduced pressure, taken out, and dried overnight in a vacuum desiccator. Take it out the next day, add 1mL DEPC water, and seal it with a stopper. Sonicate in a water bath at 50°C for 30 minutes, and pass through a 0.25 μm sterile filter to prepare cationic liposomes (CLD).

[0102] Next, the nucleic acid solution with a concentration of 20 μM is mixed with the cationic liposome according to N / P=20, and incubated at room temperature for 30 minutes, and the preparation of the cationic liposome loaded with nucleic acid is completed.

Embodiment 3

[0103] The mensuration of embodiment 3 particle size and surface potential (Zeta potential)

[0104] The invention uses Zetaszier Nano-ZS to measure and determine the particle size and surface potential of the carrier. Results The average particle size was 119.4583nm, and the particle size distribution coefficient PDI=0.224. The surface potential is 1.72e4mv. Cationic lipid carrier particle size distribution figure of the present invention is as follows figure 1 shown.

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
particle sizeaaaaaaaaaa
Login to View More

Abstract

The invention discloses a positive ion nanostructure lipid carrier, a manufacturing method and the application of the positive ion nanostructure lipid carrier. A positive ion nanostructure lipid carrier compound has a structure in a general formula VI, wherein, 3A and A4 are hydrophilic head parts, 3A or A4 is a hydrophilic molecule formed by one or multiple same or different amino acids, and 3A and A4 are the same or different; R is a linking arm between the head part and a tail part and is of a chain-shaped or branched-chain structure; and 3B and B4 are hydrophobic tail parts, 3B or B4 is a chain-shaped hydrophobic molecule, and 3B and B4 are the same or different. The positive ion nanostructure lipid carrier compound disclosed by the invention has the characteristics of simple structure, convenient composition and the like; the positive ion nanostructure lipid carrier compound also has the characteristics of low toxicity and high transfection efficiency, the complicated process in composition is avoided, auxiliary lipidosome in other types is not needed to be used in a transfection process, the operation and the use are easy, thus the positive ion nanostructure lipid carrier compound disclosed by the invention has a potential development prospect of commercial transfection reagent.

Description

technical field [0001] The invention relates to a novel non-viral gene delivery carrier, in particular to a novel cationic lipid carrier and a preparation method thereof. The invention belongs to the technical field of biomedicine, Background technique [0002] In the 1990s, human beings carried out the large-scale and extraordinary human genome project. With the completion of the whole project, human beings began their own genetic era. Human beings' further understanding of genes has also opened up the biological era of medical research, and gene therapy has naturally become one of the hot research areas of biomedicine at home and abroad. Compared with traditional therapies, gene therapy has unique advantages, especially for some hereditary or acquired diseases that cannot be cured by traditional medicine, such as some malignant tumors or innate immune diseases. A key issue in the development of gene therapy is the need for a safe and efficient gene delivery carrier. The...

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): C12N15/87
Inventor 杨振军郑宜李雅婷武芸张礼和
Owner BEIJING INTELLIGENE BIOTECH LTD CO
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