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A beta targeted recombinant lipoprotein nano drug carrier and preparation method and application thereof

A nano-drug carrier and lipoprotein technology, which is applied in the direction of drug combination, pharmaceutical formula, preparations for in vivo tests, etc., to achieve good safety effects

Inactive Publication Date: 2014-11-12
SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no research on the use of recombinant lipoprotein nano-drug delivery system for drug delivery targeting Aβ in the brain

Method used

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  • A beta targeted recombinant lipoprotein nano drug carrier and preparation method and application thereof
  • A beta targeted recombinant lipoprotein nano drug carrier and preparation method and application thereof
  • A beta targeted recombinant lipoprotein nano drug carrier and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Characterization of recombinant lipoproteins and uptake by cerebrovascular endothelial cells

[0028] (1) Preparation

[0029] Lipid (phosphatidylcholine DMPC / DOPC / DPPC+ / -ganglioside GM1+ / -cholesterol+ / -cholesterol oleate) (2-10mg), drug α-mangostin (0.1-2mg) dissolved in a certain Proportional chloroform-methanol mixed solvent, remove the organic solvent by rotary evaporation under reduced pressure, add 1ml Tris buffer (containing 10mM Tris, 0.1M KCl, 1mM EDTA, pH8.0 degassed) to the lipid film for hydration, 50°C water bath ultrasonic 1.5h . Add buffers of different pH and salt composition to dilute to 8ml, and sonicate in a water bath at 40°C. ApoE3 (0.5-5 mg) was stored in 100 mM ammonium bicarbonate solution (pH 8.0) containing 6M guanidine hydrochloride and 5 μl / (mg protein) β-mercaptoethanol. Dialyze against the appropriate protein assembly buffer before use. Add within 10 minutes, and continue to sonicate for 50 minutes. The product was cooled to ...

Embodiment 2

[0035] Example 2 Intracerebral Delivery Properties of Recombinant Lipoproteins

[0036] (1) Preparation

[0037] Weigh soybean lecithin, egg lecithin (2-10mg), ganglioside GM1 (0.1-5mg) into a 500ml round bottom flask, add 2ml of chloroform to dissolve, place in a rotary evaporator at 20°C, avoid light and vacuum for 1h. Add 2ml of PBS solution to the round bottom flask, shake at 37°C until the lipid film on the inner wall of the flask is completely hydrated and fall off, put it in a water bath at 40°C and sonicate for 1 hour, add ApoE or ApoA-I (0.1-10mg), incubate at 37°C for 36 hours, Store at 4°C.

[0038] (2) Quantitative determination of the distribution of recombinant lipoprotein in various tissues and different brain regions in the body after intravenous injection of radiolabeled

[0039] 125 I labeled recombinant lipoproteins. Twenty-four Kunming mice were randomly divided into 8 groups and administered intravenously 125 I-recombinant lipoprotein, the animals wer...

Embodiment 3

[0040] Example 3 Recombinant lipoprotein and Aβ 1-40 oligomer binding site

[0041] (1) Preparation

[0042] Weigh phosphatidylcholine and phosphatidic acid (2-10mg) into a 500ml round bottom flask, add 2ml of chloroform to dissolve, place in a rotary evaporator at 20°C, and vacuum for 1h in the dark. Add 2ml of PBS solution to the round bottom flask, shake at 37°C until the lipid film on the inner wall of the flask is completely hydrated and fall off, put it in a water bath at 40°C and sonicate for 1 hour, add ApoE or ApoA-I (0.1-10mg), incubate at 37°C for 36 hours, Store at 4°C.

[0043] (2) Observation of recombinant lipoprotein and Aβ by transmission electron microscope 1-40 oligomer binding site

[0044] 20μg / ml recombinant lipoprotein and 200nM Aβ 1-40 The oligomers were incubated in a 37°C incubator for 24 hours, centrifuged and concentrated to 800 μg / ml recombinant lipoprotein, and at the same time, the recombinant lipoprotein and Aβ 1-40 Oligomers were used as ...

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Abstract

The invention discloses an A beta targeted recombinant lipoprotein nano drug carrier and a preparation method and application thereof. The A beta targeted recombinant lipoprotein nano drug carrier comprises lipid, apolipoprotein and a drug, the drug is an alzheimer disease targeting therapeutic or diagnostic drug, and comprises one or a plurality of small molecule chemical therapeutic drugs and large molecule polypeptide, protein, gene therapeutic and diagnostic drugs. By construction of the drug carrier, the defects of scarce sources, cumbersome preparation, low quality controllability and the like of natural lipoprotein can be solved, and the drug carrier provides a new and more effective and safe solution for delivery of the alzheimer disease targeting therapeutic or diagnostic drug, and has the important research value and clinical application prospects.

Description

technical field [0001] The invention relates to the field of chemical pharmacy, in particular to an Aβ-targeted recombinant lipoprotein nano drug carrier and its preparation method and application. Background technique [0002] Alzheimer's disease (Alzheimer's Disease, AD) is the most common neurodegenerative disease, and its incidence increases with age. AD has become the third major disease that threatens the health of the elderly after cardiovascular and cerebrovascular diseases and malignant tumors. With the intensification of population aging process, the incidence of such diseases is increasing day by day. According to statistics, there are currently 35.6 million AD patients in the world, and 6 million in my country. With the aging of the population, it is estimated that by the middle of the 21st century, the number of AD patients in the world will reach 115 million. AD has become a serious threat to human health and quality of life, and is an increasingly serious pu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/42A61K48/00A61K49/00A61P25/28
Inventor 高小玲陈红专宋清香黄萌姚磊
Owner SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE
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