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Recombinant human epidermal growth factor cationic liposome and preparation method thereof

A technology of epidermal growth factor and cationic liposome, which is applied in the field of preparation and preparation of protein polypeptides, can solve the problems of short half-life, poor compatibility of cells and tissues, and low bioavailability, so as to reduce toxic side effects and improve tissue compatibility. Solubility and cell affinity, the effect of improving curative effect

Inactive Publication Date: 2013-03-06
SHENZHEN POLYTECHNIC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, EGF products on the market all have certain defects, which are mainly manifested in the following aspects: ① EGF, as a polypeptide and protein drug, has the disadvantages of poor stability, easy enzymatic hydrolysis, and short half-life of the drug in vivo. The method of repeated high-dose administration is often used in the world to overcome these shortcomings of EGF, which not only increases the inconvenience and pain of medication for patients, but also increases the cost of medication; ② EGF is easily inactive at room temperature and needs to be refrigerated at 2-8°C Preservation (rhEGF liquid preparations can only last for two months even at 4°C), which also increases the cost of storage, transportation and use of the drug; Disadvantages such as loss, low bioavailability, and poor cell and tissue compatibility

Method used

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  • Recombinant human epidermal growth factor cationic liposome and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0046] Human Epidermal Growth Factor 0.5mg

[0047] Soy Lecithin 300mg

[0048] Cholesterol 100mg

[0049] Octadecylamine 50mg

[0050] Absolute ethanol 10mL

[0051] 0.03moL / L citrate buffer 7mL

[0052] 0.1moL / L phosphate buffer 1mL

[0053] 0.5mg / mL heparin sodium solution 1mL

[0054] The preparation process is as follows: Weigh soybean lecithin, cholesterol and stearylamine according to the above prescription dosage, dissolve in 10mL of absolute ethanol, rotate and evaporate in a water bath at 50°C under reduced pressure, and pump until the absolute ethanol is evaporated to form a uniform film; Add 7mL of 0.03moL / L citric acid solution, incubate in a constant temperature water bath at 50°C, ultrasonically disperse for 10min, and granulate with a 0.22μm microporous membrane; then mix 1.0mL of rhEGF stock solution (0.5mg / mL) with 1.0mL Add heparin sodium solution after mixing, adjust the pH value to 7.2 with phosphate buffer solution, and incubate at 40°C for 10 minut...

Embodiment 2

[0058] Human Epidermal Growth Factor 0.5mg

[0059] Soy Lecithin 500mg

[0060] Cholesterol 100mg

[0061] Octadecylamine 50mg

[0062] Absolute ethanol 10mL

[0063] 0.03moL / L citrate buffer 7mL

[0064] 0.1moL / L Phosphate Buffer 3mL

[0065] 0.5mg / mL heparin sodium solution 1mL

[0066] The preparation process is as follows: Weigh soybean lecithin, cholesterol and stearylamine according to the above prescription dosage, dissolve in 10mL of absolute ethanol, rotate and evaporate in a water bath at 50°C under reduced pressure, and pump until the absolute ethanol is evaporated to form a uniform film; Add 7mL of 0.03moL / L citric acid solution, incubate in a constant temperature water bath at 50°C, ultrasonically disperse for 10min, and granulate with a 0.22μm microporous membrane; then mix 1.0mL of rhEGF stock solution (0.5mg / mL) with 1.0mL Add heparin sodium solution after mixing, adjust the pH value to 6.8 with phosphate buffer solution, and incubate at 40°C for 20 minut...

Embodiment 3

[0070] Human Epidermal Growth Factor 0.5mg

[0071] Soy Lecithin 400mg

[0072] Cholesterol 100mg

[0073] Octadecylamine 50mg

[0074] Absolute ethanol 10mL

[0075] 0.02moL / L citrate buffer 7mL

[0076] 0.1moL / L phosphate buffer 1mL

[0077] 0.5mg / mL dextran solution 1mL

[0078] The preparation process is as follows: Weigh soybean lecithin, cholesterol and stearylamine according to the above prescription dosage, dissolve in 10mL of absolute ethanol, rotate and evaporate in a water bath at 50°C under reduced pressure, and pump until the absolute ethanol is evaporated to form a uniform film; Add 7mL of 0.02moL / L citric acid solution, incubate in a constant temperature water bath at 50°C, ultrasonically disperse for 10min, pass through a 0.22μm microporous membrane to granulate; then mix 1.0mL of rhEGF stock solution (0.5mg / mL) with 1.0mL Add the dextran solution after mixing, adjust the pH value to 7.2 with a phosphate buffer solution, and incubate at 40°C for 10 minute...

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Abstract

The invention relates to a recombinant human epidermal growth factor cationic liposome and a preparation method thereof. The recombinant human epidermal growth factor cationic liposome comprises a recombinant human epidermal growth factor, a cationic liposome and a growth factor protective agent, wherein a part of the recombinant human epidermal growth factor is encapsulated by the cationic liposome. The preparation method comprises the following steps of: step 1. dissolving a panniculus adiposus material and a cationic additive by an absolute ethyl alcohol, carrying out rotary evaporation to get rid of the alcohol to form uniform panniculus adiposus, and adding a citrate buffer solution and hydrating to obtain a crude liposome suspension liquid; step 2. carrying out ultrasonic dispersion to process the crude liposome suspension liquid, and straightening particles by microfiltration membranes to obtain the blank liposome; and step 3. adding the blank liposome solution after mixing a recombinant human epidermal growth factor solution with a growth factor protective agent solution, adjusting pH to 6.0-8.0, and carrying out incubation processing to obtain the recombinant human epidermal growth factor cationic liposome. According to the preparation method, the human epidermal growth factor can be encapsulated by the cationic liposome so that the stability of rhEGF, the intermiscibility of tissues and the affinity of cells can be increased.

Description

technical field [0001] The invention relates to the technical field of preparation and preparation methods of protein polypeptides, in particular to a recombinant human epidermal growth factor cationic liposome and a preparation method thereof. Background technique [0002] Epidermal Growth Factor (EGF) is an important active protein polypeptide substance in the human body. It is an active polypeptide composed of 53 amino acids and a molecular weight of 6.2KD. Dr. Cohen, an American scientist, discovered EGF in the human body, and proved the effect of EGF on the skin, revealing the secret of human skin aging. [0003] The clinical application of EGF is mainly in: ① Accelerate the healing of wounds caused by burns, trauma, surgery, ulcers, etc., especially the healing of difficult-to-treat diabetic ulcers; ② Accelerate the healing of corneal injuries and ulcers; ③ Used for skin and corneal transplantation; ④ accelerate the healing of gastric and duodenal ulcers. In addition...

Claims

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Application Information

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IPC IPC(8): A61K9/127A61K38/18A61P17/18A61P17/02A61P27/02A61P1/04
Inventor 王金林
Owner SHENZHEN POLYTECHNIC
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