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High-efficient low-toxicity ultrasonic microvesicle for carrying ricin A chain and preparation method thereof

A technology of ricin and ultrasonic microbubbles, which can be applied to medical preparations with non-active ingredients, medical preparations containing active ingredients, echo/ultrasonic imaging agents, etc. death, etc.

Inactive Publication Date: 2012-01-04
FUJIAN MEDICAL UNIV UNION HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the lack of specificity in the binding of ricin B chain to cell membrane surface receptors, ricin often leads to the death of normal cells while systemically administering anti-tumor, resulting in serious toxic side effects

Method used

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  • High-efficient low-toxicity ultrasonic microvesicle for carrying ricin A chain and preparation method thereof
  • High-efficient low-toxicity ultrasonic microvesicle for carrying ricin A chain and preparation method thereof
  • High-efficient low-toxicity ultrasonic microvesicle for carrying ricin A chain and preparation method thereof

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Experimental program
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Effect test

Embodiment

[0025] 1. Cloning, expression and purification of ricin A chain

[0026] Extract the RNA of ricin A chain from castor beans, prepare DNA template by reverse transcription, and amplify the target gene fragment by PCR; purify the amplified gene, clone it into the PcDNA3.1 plasmid vector and perform sequence determination; sequence the correct recombinant plasmid PcDNA3.1-ricinA double digestion, insert the target fragment into the pRSetA fusion expression vector to construct the pRSetA-ricinA expression vector; transform the recombinant fusion expression vector pRSetA-ricinA into BL21 competent cells, pick clones and expand, add IPTG to induce the target gene Expressed, ultrasonically disrupted cells; purified by nickel metal chelate column, identified as ricin A chain by immunoblotting, and the half inhibitory concentration of the purified protein on liver cancer HepG2 cells was 2.0×10 as measured by MTT method for 24 hours -6 mol / L.

[0027] 2. Synthesis and characterization ...

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Abstract

The invention relates to a ricin A carried chain ultrasonic micro bubble with high efficiency and low toxicity and a preparation method thereof. The ricin A carried chain ultrasonic micro bubble is formed by combining a ricin A chain and an ultrasonic contrast agent micro bubble. The ricin A chain adopted in the combination is wrapped inside the micro bubble or adsorbed on the surface of the micro bubble. The preparation method comprises the following steps: weighing aqueous solution of the ricin A chain in proportioning amount into PLGA-PEG-PLGA solution in proportioning amount, stirring the mixture at high revolution speed to obtain primary emulsion, pouring the primary emulsion into polyvinyl alcohol aqueous solution with proper concentration and amount, stirring the mixture at high revolution speed and magnetically stirring the mixture at low revolution speed, then volatilizing organic solvent fully, centrifugalizing the mixture, washing the mixture by redistilled water to obtain the ricin A carried chain ultrasonic micro bubble, freezing and drying the ricin A carried chain ultrasonic micro bubble under vacuum, then filling in inert gas, and preserving the bubble and gas at low temperature. Therefore, through triggering the micro bubble by ultrasonic to release the ricin A chain in target fixed point, the preparation method remarkably enhances anti-tumor effect of pathologic sites, reduces toxic and side effects of whole body, and achieves the aim of target treatment of tumor.

Description

Technical field: [0001] The invention relates to ultrasonic microbubbles loaded with ricin A chain and a preparation method thereof, using ultrasonic microbubbles as a delivery tool for anti-tumor biotoxins, in particular to a novel copolymer ultrasonic microbubble loaded with ricin A chain and its preparation , the drug-loaded microbubbles can be guided by ultrasound images, and a certain energy of ultrasound can be used to rupture the drug-loaded microbubbles at the tumor site to release ricin, and enter the cells along the cell membrane with increased permeability to achieve tumor-targeted therapy the goal of. Background technique: [0002] Conventional ultrasound contrast agent is a micron-scale contrast agent, the average diameter of the microbubbles is about 2-4 μm, and it does not penetrate blood vessels. It is a blood pool imaging agent. In recent years, studies have found that microbubble ultrasound contrast agents, as a new type of drug carrier, can not only enhan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/16A61K9/00A61K47/34A61K47/42A61P35/00A61K49/22
Inventor 林礼务陈志奎薛恩生杨映红蔡敏娴杨嘉嘉
Owner FUJIAN MEDICAL UNIV UNION HOSPITAL
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