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High-throughput preparation method of therapeutic protein nanoparticles

A therapeutic protein and nanoparticle technology, applied in the field of biomedicine, can solve problems such as inapplicability, and achieve the effect of small differences between batches and large application prospects.

Active Publication Date: 2019-01-25
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, one limitation of the rapid nanoprecipitation method is that it is only suitable for hydrophobic small molecule drugs, not for biotech drugs such as protein peptides

Method used

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  • High-throughput preparation method of therapeutic protein nanoparticles
  • High-throughput preparation method of therapeutic protein nanoparticles
  • High-throughput preparation method of therapeutic protein nanoparticles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Taking insulin as a protein drug as an example, the specific method for preparing insulin nanoparticles is as follows:

[0038] (1) Hydrophobic ion pair process

[0039] ① Weigh insulin and dissolve it in a 0.01 M dilute hydrochloric acid solution, make a 5 mg / mL insulin solution, adjust the pH of the solution to 7.4 with NaOH for use;

[0040] ②Weigh DDAB and dissolve it in dichloromethane to prepare a series of DDAB organic solutions (mass concentration is 5 mg / mL~40 mg / mL);

[0041] ③After mixing equal volumes of insulin solution and DDAB dichloromethane solution (both (5 mL)), add double volume (10 mL) of ethanol to form a single-phase solution, shake at 100 rpm for 30 minutes, and add 5 mL of pure water. After phase separation and centrifugation, the lower organic phase is the solution containing the insulin-DDAB complex.

[0042] By detecting the content of insulin in the supernatant, the phase transfer rate of insulin from the aqueous phase to the organic phase is calcula...

Embodiment 2

[0061] Example 2 Hypoglycemic effect of the HA-coated insulin nanoparticle freeze-dried preparation prepared in Example 1

[0062] After dialyzing the HA-coated insulin nanoparticles prepared in Example 1, they were added with a freeze-dried protective agent and lyophilized to obtain a powder formulation, which was filled into a gelatin enteric capsule, which was the sample to be tested.

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Abstract

The invention discloses a high-throughput preparation method of therapeutic protein nanoparticles. The method comprises the following steps: firstly, converting a hydrophilic protein drug into a protein-lipid complex by using a hydrophobic ion pair method; and then diluting the protein-lipid complexes with water-soluble organic solvents, and preparing the protein nanoparticles by rapid nanoprecipitation. Compared with the traditional preparation method of protein nanoparticles, the invention has the greatest advantage that the protein nanoparticles can be continuously produced on a large scale, and the productivity can reach 1.2 g / h, which is far higher than the production capacity of the traditional preparation method of protein nanoparticles. In addition, the rapid nanoprecipitation method can be continuously produced, and the difference between batches is small, so it is suitable for industrial production. Further, the surface-coated hyaluronic acid of the protein particles preparedby the invention can impart stability and mucus penetration to the protein drug nanoparticles which can be stored for a long time. The protein nanoparticles prepared by the method of the invention have great application prospect in the aspect of in vivo delivery of protein drugs.

Description

Technical field [0001] The invention belongs to the field of biomedicine, and more specifically, relates to a high-throughput preparation method of protein nanoparticles. Background technique [0002] Biotechnology drugs mainly include peptide drugs, protein drugs and nucleic acid drugs. But so far, most of the biological drugs are protein peptides. Among the biotechnology drugs that have been marketed and developed, protein drugs account for the vast majority, including cytokine drugs and recombinant hormone drugs. At present, protein peptide drugs play an irreplaceable role in tumors, autoimmune deficiency diseases and cardiovascular diseases because of their high specificity and excellent curative effect. [0003] However, compared with conventional drugs, protein peptide drugs have large molecular weight, poor stability, variability, and short half-life in the body, which makes it difficult to penetrate the intestinal mucosa, which limits their wide application. In recent yea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/51A61K47/24A61K38/28A61K38/38A61K47/10A61K47/18A61K9/19C07K1/30
CPCA61K9/19A61K9/5161A61K38/28A61K38/38A61K38/385A61K47/10A61K47/183A61K47/24C07K1/30
Inventor 曾志鹏陈永明高虓虎刘利新赵鹏飞
Owner SUN YAT SEN UNIV
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