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Hybrid-type multi-lamellar nanostructure of epidermal growth factor and liposome and method for manufacturing same

A technology of epidermal growth factor and nanostructure, which is applied in liposome delivery, skin care preparations, medical preparations containing active ingredients, etc., can solve the problems of low encapsulation efficiency of active ingredients and decreased protein physiological activity, etc., and achieve The effect of simple manufacturing process and high encapsulation efficiency

Active Publication Date: 2017-08-29
CELLTRION INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the liposomes containing physiologically active proteins disclosed in the patent documents described above have the following problems: the encapsulation efficiency of the active ingredient is very low, and since the liposomes are prepared under high temperature and pressure, the physiological activity of the protein severe decline

Method used

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  • Hybrid-type multi-lamellar nanostructure of epidermal growth factor and liposome and method for manufacturing same
  • Hybrid-type multi-lamellar nanostructure of epidermal growth factor and liposome and method for manufacturing same
  • Hybrid-type multi-lamellar nanostructure of epidermal growth factor and liposome and method for manufacturing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1: Preparation of EGF-DOTAP hybrid multilayer nanostructure

[0063] 1.1: Preparation of cationic empty monolamellar liposomes containing DOTAP

[0064] The cationic lipid DOTAP (20.96 mg, Avanti Polar Lipid, Inc.) was dissolved in 1 ml of chloroform and mixed in a round glass flask. Chloroform was removed by flushing the lipid solution with nitrogen gas at low speed in a rotary evaporator, and the lipid was dried to form a thin lipid layer. The formed lipid layer was further dried in vacuum for 12 h to completely remove the remaining chloroform. 1 ml of purified water was added to the prepared lipid layer, followed by stirring at 37°C for 2 hours, thereby preparing empty lipid vesicles. The obtained empty lipid vesicles were extruded through a polycarbonate membrane (polycarbonate membrane) (Avanti Polar Lipid, Inc.) with a pore size of 100 nm (Avanti Polar Lipid, Inc.) several times to prepare a cationic empty monolayer lipid containing DOTAP and havi...

experiment Embodiment 1

[0067] Experimental Example 1: Evaluation of Formation of EGF-DOTAP Hybrid Multilayer Nanostructures

[0068] 1.1: Confirmation of the formation of cationic empty unilamellar liposomes

[0069] The particle size and zeta potential of the cationic empty unilamellar liposomes prepared in Example 1.1 were measured using dynamic light scattering (DLS, ELSZ-1000, Otsuka Electronics), and the measurement results are shown in Table 1 below. The measurement results showed that the prepared cationic empty monolamellar liposome had a particle size of 200nm and a positive surface charge.

[0070] [Table 1]

[0071] Empty unilamellar liposomes

[0072] Empty unilamellar liposomes

Particle size (nm)

Zeta potential (mV)

DOTAP

197.7±4.9

56.5±2.5

[0073] 1.2: Confirmation of the formation of EGF-DOTAP hybrid multilayer nanostructures

[0074] Use DLS to measure the particle size and zeta potential of the EGF-DOTAP hybrid multilayer nanostructure p...

experiment Embodiment 2

[0077] Experimental Example 2: Evaluation of EGF Encapsulation Efficiency in EGF-DOTAP Hybrid Multilayer Nanostructures

[0078] In order to measure the amount of EGF encapsulated in the EGF-DOTAP hybrid multilayer nanostructures, the EGF-DOTAP hybrid multilayer nanostructures prepared in Example 1.2 were subjected to an ultracentrifuge (200,000 x g, 2 hours, 4°C, Beckman). Layer nanostructures (1 ml) were centrifuged to separate unencapsulated free EGF. The amount of isolated free EGF was measured using a micro BCA assay and an ELISA assay. The measurement results are shown in Figure 4 .

[0079] From Figure 4 As can be seen in , high encapsulation efficiencies of 60% or higher occur at most EGF / DOTAP weight ratios, which vary slightly depending on the method used to quantify EGF. Thus, it can be seen that the encapsulation efficiency in the structures according to the invention is significantly higher than in conventional liposomes (only 10-20%).

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Abstract

The present invention relates to a hybrid-type multi-lamellar nanostructure of an epidermal growth factor and a liposome and a method for manufacturing same. The new kind of hybrid-type multi-lamellar nanostructure not only has a high epidermal growth factor sealing efficiency, but also can be manufactured through a simple process, such that the same can be easily delivered into a living body or a cell while maintaining a high physiological activity of the epidermal growth factor.

Description

technical field [0001] The invention relates to a mixed multilayer nanostructure of epidermal growth factor (EGF) and liposome and a manufacturing method thereof. More specifically, the present invention relates to a novel protein-lipid hybrid multi-lamellar nanostructures (protein-lipid hybrid multi-lamellar nanostructures) and its manufacturing method, the protein-lipid hybrid multi-lamellar nanostructures are It is made by multivalent electrostatic interactions (multivalent electrostatic interactions) in addition to hydrophobic interactions between empty vesicles formed by plasmin and anionic epidermal growth factor proteins. Background technique [0002] Due to growing interest in cosmetology, physiologically active proteins having specific action mechanisms and good efficacy are attracting attention as materials for functional cosmetic products. However, physiologically active proteins do not easily penetrate the skin due to their high molecular weight, short half-life...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K8/14A61K8/64A61Q19/00
CPCA61K8/14A61K8/64A61Q19/00A61K8/553A61Q19/08A61K2800/56A61K9/0014A61K9/127A61K9/1272A61P17/00
Inventor 南允盛赵诚德具本一
Owner CELLTRION INC
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