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Preparation technology and application of natural recombinant nanostructured lipid carrier

A natural and lipid technology, applied in drug combination, freeze-dried delivery, powder delivery, etc., can solve the problems of total protein miscellaneous, unfavorable waste utilization, low economic value, etc.

Active Publication Date: 2015-02-04
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the method described in this invention has a high protein yield, the protein is always miscellaneous and many are unfavorable for the utilization of waste in plasma production
[0004] CN102127165B discloses a method for purifying and obtaining high-purity apoA-I from plasma fraction IV, through the following steps: plasma fraction IV is precipitated and dissolved, centrifuged to remove diatomaceous earth and impurities, and supernatant is collected; supernatant is added with chlorine The sodium chloride solution is centrifuged to obtain the apoA-I precipitate; the precipitate is redissolved and filtered; the filtrate is successively separated by anion column chromatography and hydrophobic column chromatography to obtain a high-purity apoA-I solution, but the operation process of the invention is complicated and the economic value is low

Method used

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  • Preparation technology and application of natural recombinant nanostructured lipid carrier
  • Preparation technology and application of natural recombinant nanostructured lipid carrier
  • Preparation technology and application of natural recombinant nanostructured lipid carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Extraction and purification process of natural apolipoprotein and lipid in plasma fraction IV

[0042] The invention provides a natural apolipoprotein extraction and purification process, comprising the following steps:

[0043] Utilize organic solvent combined with isoelectric point precipitation method, the steps of extracting apolipoprotein mixture from human plasma fraction IV are as follows:

[0044] Step 1) Weigh plasma fraction IV, initially dissolve in 50ml of phosphate buffer solution with pH 5.25, add 16.6ml of 25% ethanol solution to continue dissolving for an appropriate time, and centrifuge at 12000r / min for 15min.

[0045] Step 2) Take the precipitate, initially dissolve it in 50ml of phosphate buffer solution with pH 6.6, add 4.4ml of 8% ethanol solution to continue dissolving for an appropriate time, and centrifuge at 12000r / min for 15min.

[0046] Step 3) Take the supernatant, adjust the pH to 5.25 with dilute hydrochloric acid, add 12.5ml o...

Embodiment 2

[0050] Embodiment two: the preparation process of carrying PTX natural recombinant lipoprotein (natural recombinant lipoprotein-PTX) nanoparticle

[0051] 1.1 Preparation of natural recombinant lipoprotein-PTX nanoparticles by emulsification evaporation method

[0052]

[0053] The natural lipoprotein mixture and lipids were obtained by organic solvent combined with isoelectric point precipitation method, and the natural lipid ethanol solution dissolved in PTX was slowly added drop by drop with a syringe under the action of magnetic stirring at room temperature to the phosphate buffer solution of apolipoprotein , after the dropwise addition, continue magnetic stirring for 50 min; remove ethanol by rotary evaporation at 37°C, and conduct probe ultrasound under ice bath conditions, pass through a 0.22 μm filter membrane, and freeze-dry to obtain natural recombinant lipoprotein-PTX nanoparticles.

[0054] 1.2 Preparation of natural recombinant lipoprotein-PTX nanoparticles by ...

Embodiment 3

[0060] Example 3: Research on the properties of natural recombinant lipoproteins

[0061] 2.1 Particle size and morphology of natural recombinant lipoprotein-PTX nanoparticles

[0062] The average particle size of natural recombinant lipoprotein-PTX nanoparticles was measured by laser particle size analyzer (160.4±20)nm. Transmission electron microscope results are as figure 1 As shown, the prepared natural recombinant lipoprotein-PTX nanoparticles are all spherical particles with good roundness, smooth appearance, and consistent with the particle size measured by the laser particle size analyzer.

[0063] 2.2 Encapsulation efficiency of PTX in natural recombinant lipoprotein-PTX nanoparticles

[0064] Take 100 μl of natural recombinant lipoprotein-PTX nanoparticles, dilute to 5ml with methanol, sonicate in a water bath for 20 minutes, centrifuge at 12000r / min for 10 minutes, take the supernatant and filter it with a 0.22 μm organic filter membrane, and measure its content b...

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Abstract

The invention belongs to the field of biological medicinal preparation and relates to a preparation technology and an application of a natural recombinant nanostructured lipid carrier. The specifically-refereed nanoparticle is natural recombinant lipoprotein drug-loaded nanoparticle and is prepared by in vitro recombination of an apolipoprotein mixture and lipid which are obtained by extraction and purification of a blood plasma ingredient IV and a medicament. The recombination process is realized by a film dispersion method, an emulsification-evaporation method or an emulsification-evaporation improved method. The technical problem to be solved in the invention is the extraction and purification method of apolipoprotein and lipid mixture and the preparation technology of the natural recombinant lipoprotein drug-loaded nanoparticle. Preparation conditions are mild, and costs are low. The natural recombinant lipoprotein nanoparticle provided by the invention has remarkable advantages of spheroidic structure, high bionic performance, high penetrability, efficient carrying performance, tumor targeting and the like. The invention provides property evaluation and application of the above natural recombinant lipoprotein nanoparticle. Normal saline, or a phosphate buffer or 5% glucose solution is added to dissolve the nanoparticle, and the dissolved nanoparticle can be used to treat diseases in a mode of intravenous injection, intramuscular injection, oral medication or cutaneous penetration. The technology provided by the invention is simple and low-cost and is easy for industrial production.

Description

technical field [0001] The invention relates to the field of biopharmaceutical preparations, in particular to natural recombinant lipoprotein nanoparticles for tumor targeting and treatment and a preparation process thereof. Background technique [0002] Lipoproteins are a type of heterogeneous particles with differences in particle size, lipid composition, and apolipoprotein types. They are biomacromolecules composed of a phospholipid monolayer containing apolipoproteins and free cholesterol and a non-polar lipid core. , which play a key role in lipid transport in vivo. According to the density, plasma lipoproteins (Table 1) can be divided into four types by density ultracentrifugation: chylomicrons (Chylomicrons, CM), very low-density lipoprotein (VLDL), low-density lipoprotein (Low-density lipoprotein, LDL) and high-density lipoprotein (High-density lipoprotein, HDL). Since there is a lipid core in the lipoprotein structure, and LDL and HDL can be endocytosed and absorb...

Claims

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Application Information

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IPC IPC(8): A61K9/14A61K9/19A61P35/00
Inventor 周建平王若宁丁杨王伟
Owner CHINA PHARM UNIV
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