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, microorganism-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

[0084] [Example 1] Preparation of oleZH 2 oil body

[0085] according to make figure 2 The preparation process shown constructs the oleZH of the present invention 2 oil body.

[0086]

[0087] The following three genes are constructed on the expression vector by gene recombination technology:

[0088] Oil body membrane protein (N-terminal)-Z Her2 Peptide (C-terminal) oleZH 2 The gene of the protein: a linker (comprising the amino acid sequence listed in SEQ ID NO:4) with a nucleotide sequence listed in SEQ ID NO:3 combined with the oil body membrane protein gene of sesame seeds (comprising SEQ ID NO:5 Nucleotide sequence listed) and ligand peptide of HER2 / neu protein (i.e. Z Her2 peptide) gene (comprising the nucleotide sequence listed in SEQ ID NO: 6). The detailed operation steps are as follows. First, Nord et al. developed a ZHer2 peptide that can specifically bind to the HER2 / neu protein receptor as a ligand for the receptor, see Nord et al., Binding proteins se...

Embodiment 2

[0096] [Example 2] oleZH 2 Atomic Force Microscopy Testing of Oil Bodies

[0097] The mica sheet (purchased from Youhong Enterprise Co., Ltd.) was washed with water three times for three minutes each time, soaked with 0.02M MgCl for 2-3 minutes, and washed once with water. The oleZH prepared in step 2 2 Protein (50 mg) was added to a test tube and 50 mg triglycerides, 150 μg phospholipids were added and assembled into oleZH 2 Oil body, let the oil body pass through a 0.22mm filter membrane, 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, and then wash it twice with double distilled water , placed in a 37°C oven to dry, and finally observed the shape and size of the oil body with an atomic force microscope (Atomic Force Microscope, AFM) (model: NS4 / D3100CL / MultiMode), the results are as follows Figure 7 shown.

Embodiment 3

[0098] [embodiment 3] lipid and oleZH 2 Effect of protein ratio

[0099] In 100 micrograms oleZH 2 \protein(pJO1-oleZH 2 ) in 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 obtain oleZH-containing 2 Protein / lipid (olive oil) weight / volume ratio (µg / µl) 2 / 1 mixture. Then, ultrasonically shake the mixture (the mixture is placed on ice, power: 15%, time: 20 seconds, run: 0.5 seconds, rest: 0.5 seconds, shake three times) to obtain the desired oleZH 2 oil body. Repeat the previous operation, but use 400, 200, 100 and 100 micrograms of oleZH 2 protein, and correspondingly use 20, 20, 100 and 500 microliters of olive oil to obtain oleZH 2 oleZH with protein / lipid (olive oil) weight / volume ratios (µg / µl) of 20 / 1, 10 / 1, 1 / 1, and 1 / 5, respectively 2 oil body.

[0100] Using Nikon 104 optical microscope to observe different conditions oleZH 2 The shape...

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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: an oleZH2 protein, wherein the oleZH2 protein comprises oleosin and a ZHer2 peptide; a lipid, a lipid-coated liposoluble drug, and a lipid-coated known fluorescence molecule, wherein the materials are prepared into an oleZH2 oil body, an oleZH2 drug oil body and an oleZH2 fluorescence oil body so as to provide an active targeted drug delivery system and a detection system.

Description

technical field [0001] The present invention utilizes oil body membrane protein to connect Z Her2 peptides, which assemble into oleZH 2 protein, this oleZH 2 Proteins can coat lipids, fat-soluble drugs or known fluorescent signal molecules to become oleZH 2 Oil body, then oleZH 2 Oil body is applied to the application of target therapy or detection; The present invention utilizes oil body membrane protein to connect a specific linker (linker) to connect Z Her2 Peptides, Z can be purified by separation Her2 Protein, and get high purity oleZH 2 Oil body; the present invention utilizes exonuclease peptide or endonuclease peptide to connect oil body membrane protein to connect Z Her2 Peptides are then linked to exonuclease peptides or endonuclease peptides to assemble into oleZH 2 oil body, the oleZH 2 Oil bodies can detect cancerous sites and enter overexpressed cancer cells to cause cell division or self-apoptosis. Background technique [0002] According to statistics...

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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