Nano-scale artificial oil body for targeted drug delivery system detection and treatment

A delivery system, nano-scale technology, applied in drug delivery, drug combination, microbial-based methods, etc., can solve problems such as limited purification technology

Inactive Publication Date: 2012-10-24
陈致融
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of antibody immobilized immunoaffinity techniques is also limited for purification techniques

Method used

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  • Nano-scale artificial oil body for targeted drug delivery system detection and treatment
  • Nano-scale artificial oil body for targeted drug delivery system detection and treatment
  • Nano-scale artificial oil body for targeted drug delivery system detection and treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] [Example 1] Preparation of calEGFR oil body

[0089] according to make figure 2 The preparation scheme shown constructs the calEGFR oil bodies of the present invention.

[0090]

[0091] The following three genes are constructed on the expression vector by gene recombination technology:

[0092] Calpain (N-terminus)-EGFR peptide (C-terminus), gene of calEGFR protein: a gene of calpain from sesame seeds (comprising the nucleotide sequence listed in SEQ ID NO: 3) combined with the gene of EGFR protein The gene of the ligand peptide (ie EGFR peptide) (comprising the nucleotide sequence listed in SEQ ID NO:4). The detailed operation steps are as follows. First, use sesame seeds for gene recombination, purification and extraction, and obtain calpain peptide) to assemble pET-calpain peptide, purify pET-calpain peptide as template DNA, and then use primers An oil body calpain gene fragment with a size of 748bp was obtained by PCR. Next, digest the gene fragment with Nd...

Embodiment 2

[0100] [Example 2] Atomic force microscope test of calEGFR oil body

[0101] Rinse the mica sheet (purchased from Youhong Enterprise Co., Ltd.) three times with water for three minutes each time, soak it with 0.02M MgCl for 2-3 minutes, and wash it once with water. Add the calEGFR protein (50 mg) prepared in step 2 to a test tube, add 50 mg triglycerides, 150 micrograms of phospholipids and assemble into calEGFR oil bodies, and pass the oil bodies through a 0.22 mm filter membrane , and then add the filtered oil body to the mica sheet and let it stand for 2 hours to make the targeted artificial oil body adsorb on the mica sheet, then wash it twice with double distilled water, place it in a 37°C oven to dry, and finally use atomic force Microscope (Atomic Force Microscope, AFM) (Model: NS4 / D3100CL / MultiMode) to observe the shape and size of oil bodies, the results are as follows Figure 7 shown.

Embodiment 3

[0102] [Example 3] The influence of the ratio of lipid and calEGFR protein

[0103] Add 950 microliters of sodium phosphate buffer (0.01 molar concentration, pH 7.5) (purchased from Sigma Company) and 50 microliters of olive oil (purchased from Sigma Company) to 100 micrograms of calEGFR\protein (pET20bI-calEGFR), to obtain Contains a 2 / 1 mixture of calEGFR protein / lipid (olive oil) at a weight / volume ratio (µg / µl). Then, the mixture was ultrasonically oscillated (the mixture was placed on ice, power: 15%, time: 20 seconds, run: 0.5 seconds, rest: 0.5 seconds, oscillated three times) to obtain the desired calEGFR oil body. Repeat the previous operation, but use 400, 200, 100 and 100 micrograms of calEGFR protein respectively, and use 20, 20, 100 and 500 microliters of olive oil correspondingly, to obtain the weight / volume ratio of calEGFR protein / lipid (olive oil) ( μg / μl) were 20 / 1, 10 / 1, 1 / 1, and 1 / 5 calEGFR oil bodies, respectively.

[0104] A Nikon 104 optical microscope...

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Abstract

The present invention relates to a nano-scale artificial oil body for targeted drug delivery system detection and treatment. The nano-scale artificial oil body comprises: a calEGFR protein, wherein the calEGFR protein comprises caleosin and a EGFR peptide; a lipid, a lipid-coated liposoluble drug, and a lipid-coated known fluorescence molecule, wherein the materials are prepared into a calEGFR oil body, a calEGFR drug oil body and a calEGFR fluorescence oil body so as to provide an active targeted drug delivery system and a detection system.

Description

technical field [0001] The present invention relates to a targeted artificial oil body, which involves a receptor on the surface of cancer cells that can be accurately recognized by the EGFR peptide in the calEGFR protein, and is made with lipids, fat-soluble drugs and or known fluorescent signal molecules calEGFR drug oil body or calEGFR detection oil body, to achieve a nano-scale artificial detection and treatment of targeted drug delivery system that can specifically identify cancerous sites and self-assemble drug proteins for cancerous site detection, inhibition and drug treatment oil body. Background technique [0002] Cancer is currently the main killer that threatens human life and health around the world. According to the statistics of the American Cancer Society, in 2007, about 7.6 million people died of various types of cancer in the world. According to statistics from the Taiwan Department of Health, about 38,913 people in Taiwan died of malignant tumors in 2008....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/00A61K47/42A61K47/44A61K49/00C12N15/63C12N15/70A61P35/00C12R1/19
CPCA61K31/475A61K47/44A61K38/00A61P35/00Y02A50/30
Inventor 陈顺基朱宝美
Owner 陈致融
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