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

Method for massively preparing midkine antisense oligodeoxynucleotide nano-liposomes for injection

An antisense oligonucleotide and nanoliposome technology, applied in liposome delivery, gene therapy, pharmaceutical formulations, etc., can solve problems such as instability and difficulty in passing through cell membranes, and achieve the effect of improving curative effect

Active Publication Date: 2013-03-27
HUZHOU CENT HOSPITAL
View PDF2 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the antisense oligonucleotide is a highly hydrophilic substance with a strong negative charge on the surface, it is difficult to pass through the cell membrane, and it is easily degraded by various enzymes in the blood and is unstable. Suitable gene delivery vectors deliver antisense oligonucleotides to target cells

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
  • Method for massively preparing midkine antisense oligodeoxynucleotide nano-liposomes for injection
  • Method for massively preparing midkine antisense oligodeoxynucleotide nano-liposomes for injection
  • Method for massively preparing midkine antisense oligodeoxynucleotide nano-liposomes for injection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1: Preparation of 100ml injection midkine antisense oligonucleotide nanoliposomes

[0040] Raw material: DC-Chol 100mg

[0041] DOPE 100mg

[0042] MK-ASODN 5.5g

[0043]Methanol + absolute ethanol 5ml

[0044] Glucose 8g.

[0045] Preparation:

[0046] (1) Stir DC-Chol and DOPE with methanol: absolute ethanol (v / v=1:1) at a temperature of 45°C until the solution is clear and transparent to obtain a lipid solution, which is then kept at 45°C;

[0047] (2) Then use cross-flow technology to quickly mix the lipid solution and water in a certain proportion, the volume ratio is less than 3:17, that is, the concentration of the volume percentage is lower than 15%, and the liposome obtained after mixing is ultrasonicated by a probe-type ultrasonic instrument (power 200w, ultrasonic 100 times, ultrasonic 1s, intermittent 2s), and then through a high-pressure infusion pump, the liposomes were extruded 5 times through an extruder covered with a 0...

Embodiment 2

[0051] Embodiment 2: Preparation of 500ml injection midkine antisense oligonucleotide nanoliposomes

[0052] Raw material: DC-Chol 500mg

[0053] DOPE 36.44mg

[0054] MK-ASODN 2.79mg

[0055] Absolute ethanol 10ml

[0056] Mannitol 25g.

[0057] Preparation:

[0058] Mix DC-Chol and DOPE dropwise with absolute ethanol at a temperature of 20°C until the solution is clear and transparent to obtain a lipid solution, then use cross-flow technology to quickly mix the lipid solution and phosphate buffer in a certain proportion, and mix Finally, the liposomes obtained were ultrasonicated in a water bath (power 120w, ultrasonic 5min), and then the liposomes were extruded twice through an extruder with five 0.1μm polycarbonate membranes to a uniform particle size through a high-pressure infusion pump. The liposome suspension that flows out passes through ultrafilter ultrafiltration immediately, removes organic solvent, and the liposome hydration liquid is ...

Embodiment 3

[0060] Embodiment 3: Preparation of 1000ml injection midkine antisense oligonucleotide nanoliposomes

[0061] Raw material: DC-Chol 1000mg

[0062] DOPE 72.88mg

[0063] MK-ASODN 5.58mg

[0064] Methanol 20ml.

[0065] Preparation:

[0066] Stir DC-Chol and DOPE with absolute ethanol at a temperature of 40°C until the solution is clear and transparent to obtain a lipid solution, and then use cross-flow technology to rapidly mix the lipid solution and Tris buffer in a certain proportion, and mix Finally, the obtained liposomes were ultrasonicated in a water bath (power 120w, ultrasonic 10min), and then through a high-pressure infusion pump, the MK-ASODN liposomes passed through an extruder covered with 10 pieces of 0.2μm polycarbonate membranes to achieve uniform particle size; The outflowing liposome suspension is immediately ultrafiltered through an ultrafilter to remove organic solvents, and Tris buffer is continuously replenished in the middle, which is about 6 times th...

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
pore sizeaaaaaaaaaa
particle diameteraaaaaaaaaa
Login to View More

Abstract

The invention relates to a method for massively preparing midkine antisense oligodeoxynucleotide nano-liposomes for injection. The method comprises the following steps: dissolving a mixture of cationic liposomes and lecithoid liposomes in an organic solvent to obtain a liposome liquor; then, initially dispersing the liposomes by an ultrasonic instrument, and ultrafiltering to remove the organic solvent, and finally squeezing by a squeezer at a high pressure to uniform grain size to obtain blank nano-liposomes; and then, mixing with an MK-ASODN liquor to obtain MK-ASODN nano-liposomes. The grain size of the midkine antisense oligodeoxynucleotide nano-liposomes for injection produced by the method is within 100-500nm, and the midkine antisense oligodeoxynucleotide nano-liposomes for injection is stable in property and the anti-hepatoma effect of the midkine antisense oligodeoxynucleotide nano-liposomes for injection is remarkably enhanced. The method is simple in process, easy to realize, very easy to amplify, and suitable for industrial production.

Description

[0001] technical field [0002] The invention belongs to the technical field of preparation of anti-liver cancer drugs, and relates to a method for large-scale preparation of midkine antisense oligonucleotide nano-liposomes. [0003] Background technique [0004] Primary hepatocellular carcinoma (hepatocellular carcinoma, HCC) is a highly invasive, highly metastatic solid malignant tumor, but there is still a lack of ideal treatment. [0005] Midkine (MK) is a newly discovered pro-angiogenic factor. Basic heparin-binding protein expressed only in mid-embryonic and adult kidneys. Existing studies have shown that MK mRNA and protein are highly expressed in many human tumor tissues such as liver cancer, while various normal tissues have only low or no expression, and are related to the occurrence, invasion and metastasis, angiogenesis and prognosis of liver cancer. are closely related, and angiogenesis is the basis for unrestricted invasion, growth and metastasis of liver ca...

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): A61K48/00A61K9/127A61P1/16A61P35/00
Inventor 戴利成钟婧
Owner HUZHOU CENT HOSPITAL
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