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A kind of preparation method of strychnine immune nanoparticles

A strychnine and nanoparticle technology, which is applied in the directions of non-active ingredients medical preparations, medical preparations containing active ingredients, pharmaceutical formulas, etc., can solve the problems of large dosage and strong toxic and side effects, and achieve drug release. Stable, good biocompatibility, good spheroidizing effect

Inactive Publication Date: 2011-12-07
SHANGHAI UNIV OF TRADITIONAL CHINESE MEDICINE PUTUO DISTRICT CENT HOSPITAL +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, strychnine is a highly toxic traditional Chinese medicine, and its therapeutic dose is very close to the poisoning dose. The systemic drug dose is large, and its toxic and side effects are strong.

Method used

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  • A kind of preparation method of strychnine immune nanoparticles
  • A kind of preparation method of strychnine immune nanoparticles
  • A kind of preparation method of strychnine immune nanoparticles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Preparation method of anti-human AFP McAb-polyethylene glycol-polylactic acid block copolymer strychnine immune nanoparticles

[0048] The raw material formula is as follows:

[0049]

[0050] Among them, the polymer material (PLA-PEG-CH2-CH2-COOH) is a carboxylated polyethylene glycol formed by copolymerization of carboxylated polyethylene glycol and polylactic acid at a weight ratio of 1:1.5~3 (preferably 1.5:2.5). Diol-polylactic acid block copolymer; its general formula is PLA-PEG-CH2-CH2-COOH, its molecular weight range is 40-50kD, its structural fragments and 1 H-NMR measurement spectrum such as figure 1 shown.

[0051] Mix the above-mentioned oil phase and water phase, disperse with a high-shear homogenizer, shear at 12000 rpm for 5 minutes to obtain a dispersion, and then continue to emulsify the dispersion under 300w power ultrasonic conditions, ultrasonic 5 times, each 30s, Get first emulsion. Pour the primary emulsion into the diluent of the prescripti...

Embodiment 2

[0055] In vitro drug release test of anti-human AFP McAb-polyethylene glycol-polylactic acid block copolymer strychnine immune nanoparticles

[0056] Precisely pipette 2.0ml of strychnine immune nanoparticle concentrate, place it in a dialysis bag with a molecular weight cut-off of 3,000 Daltons, tie both ends tightly, operate 3 parts in parallel, and put it into 50ml of PBS medium with pH7.4 In vitro release at 37°C and 100rpm rotation speed, 4ml samples were taken at 0.5, 1, 2, 4, 8, 12, 16, 18, 24, 36, 48h after the start of the release, 4ml of rehydration solution was taken, and the samples were released at a wavelength of 263nm Determine the UV absorption value, calculate the concentration of Bru (strichnine) in the release medium and the cumulative release percentage.

[0057] Another Bru-PBS solution with a concentration of 0.427 mg / ml was prepared, 2.0 ml of the solution was precisely pipetted, placed in a dialysis bag with a molecular weight cut-off of 3,000 Daltons, ...

Embodiment 3

[0060] Determination of targeting of anti-human AFP McAb-polyethylene glycol-polylactic acid block copolymer strychnine immune nanoparticles

[0061] Anti-human AFP McAb strychnine immune nanoparticles were incubated with liver cancer cells SMMC-7721 for 4 h, and the cells were washed 3 times with 0.01M PBS; FITC-labeled secondary antibody was added, incubated for 2 h, and washed 3 times with PBS; Complementary immunofluorescence methods were observed under a confocal microscope. A 200X confocal microscope showed that strychnine immune nanoparticles were evenly distributed near the liver cancer cell membrane, showing an approximate "ring" shape, showing good targeting, as shown in Figure 5 Shown (photograph under confocal microscope × 200 times).

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Abstract

The invention relates to a preparation method of strychnine immune nanoparticles, and the preparation method comprises the following specific steps: step 1, dissolving carboxylated poly(ethylene glycol)-polylactic acid block copolymer and strychnine in an organic solvent, and then mixing with a polyvinyl alcohol aqueous solution to form a primary emulsion; step 2, removing the organic solvent andimpurities to obtain a strychnine nanoparticle concentrate; and step 3, sequentially adding a carbodiimide salt used as a carboxyl activation reagent and an anti-human-AFP (alpha-fetoprotein) monoclonal antibody to the strychnine nanoparticle concentrate, thus linking the carboxyl group on polyethylene glycol and the amino group of the anti-human-AFP monoclonal antibody by chemical coupling to obtain the strychnine immune nanoparticles. The preparation method provided by the invention has the advantages of simple process, high encapsulation rate, stable drug release and good drug ballability;and the obtained strychnine immune nanoparticles are used for preparing anti-tumor immune targeting drugs, and have the advantages of precise targeting drug accumulation in tumor tissue cells, stabledrug release, good anti-cancer effect, safety and the like.

Description

technical field [0001] The present invention relates to a preparation method of strychnine immune nanoparticles, in particular, to a liver cancer-specific targeted therapy drug—anti-human AFP McAb (anti-human alpha-fetoprotein monoclonal antibody)-polyethylene glycol - Preparation method of polylactic acid block copolymer strychnine immune nanoparticles. Background technique [0002] Strychnine (Brucine) is one of the main components of Strychnium, and its chemical structure is as follows: [0003] The strychnine is a weakly basic alkaloid with poor water solubility. As a representative drug for promoting blood circulation and unblocking collaterals and "fighting poison with poison", it can promote the proliferation of T lymphocytes, and has a dose-dependent functional regulation of lymphocyte function in mice. It has obvious tumor inhibitory effect and strong analgesic effect. Deng Xukun et al. evaluated the effect of strychnine on the tumor inhibitory effect and surviva...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/14A61K31/475A61K47/34A61K47/42A61K47/48A61P35/00
Inventor 秦建民盛霞杨林撒忠秋黄涛李琦殷佩浩张敏高科攀陈庆华马经纬沈鹤柏
Owner SHANGHAI UNIV OF TRADITIONAL CHINESE MEDICINE PUTUO DISTRICT CENT HOSPITAL
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