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Preparation method of artificial liposome

A liposome and artificial technology, which is applied in the field of pharmacy, can solve the problems of long process cycle, harsh conditions, complicated preparation process of liposome products, etc., and achieve the advantages of simple separation process, mild conditions, and prevention of configuration changes and activity decline. Effect

Inactive Publication Date: 2012-07-11
LULIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the few research reports, the preparation process of modified or encapsulated liposome products is complicated, the conditions are harsh and the process cycle is long

Method used

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  • Preparation method of artificial liposome
  • Preparation method of artificial liposome
  • Preparation method of artificial liposome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Preparation of bitter gourd protein MAP30 and egg yolk lecithin zirconium Zr (EPC) 2 Composite liposomes: Momordica charantia protein (from seeds) crude extract, confirmed by SDS-PAGE gel electrophoresis with a molecular weight of 30 kDa, accurately weighed 6.8 mg MAP30 and dissolved in 1.0 ml phosphate buffer (pH 7.2, 10 mM + 100 mM NaCl) for future use; egg yolk lecithin zirconium nanocarriers (5nm, spherical particles) were prepared according to the method in the patent (CN102107131A). Accurately weigh 201.6 μg of Zr(EPC)2 particle carrier, add 1.49 ml of phosphate buffer (pH 7.2), stir at a high speed (3500 rpm) at 4 oC until the carrier is completely suspended and dispersed, then add 0.01 ml of MAP30 protein stock solution , stirred for 0.5, 1.0, 4.0, 7.0, 10.0, 12.0, 15.0, 18.0, 20.0 and 24.0 hours respectively, and separated the supernatant after high-speed centrifugation (12000 rpm) for 20 minutes. The solid particles were washed with 0.1ml phosphate buffer sol...

example 1

[0032] Example 1 artificial liposome product

[0033] The product was freeze-dried for 3 days: white powder, insoluble in water and polar organic solvents; the adsorption time corresponding to the maximum protein content was 15 hours, the optimum pH was 8.52, and the maximum percentage of protein was 24.0 ± 1.0%; After dispersing in phosphate buffer solution, a colorless transparent suspension is obtained; under solid phase conditions, the thermal desorption temperature of protein is 80 oC (within 5 hours); under simulated physiological conditions in vitro, liposomes are stable (protein desorption rate ≤ 5%) for up to 9 months; it is used for the construction and development of stable dosage forms of ribosome-inactivating high-efficiency anticancer drugs, which can be used for the treatment of type I HIV.

[0034] figure 2 Zirconium in egg yolk lecithin (Zr(EPC) for bitter melon protein drug MAP30 2 ) Experimental results of adsorption kinetics of nanocarriers; the experime...

Embodiment 2

[0036] Preparation of balsam pear protein MAP30 and soybean lecithin zirconium Zr (SPC) 2 Composite liposomes: Accurately weigh 6.8 mg of bitter melon protein, dissolve in 1.0 ml phosphate buffer (pH 7.2, 10 mM + 100 mM NaCl) and save for later use; soybean lecithin zirconium nanocarriers (5nm, spherical particles) are patented (CN102107131A) prepared by the method. Accurately weigh 201.6 μg Zr(SPC) 2 For particle carrier, add 1.49 ml phosphate buffer (pH 7.2), stir at high speed (3500 rpm) at 4 oC until the carrier is completely suspended and dispersed, then add 0.01 ml MAP30 protein stock solution, stir 0.5, 1.0, 4.0, 7.0 respectively , 10.0, 12.0, 15.0, 18.0, 20.0 and 24.0 hours, after high-speed centrifugation (12000 rpm) for 20 minutes, the supernatant was separated. The solid particles were washed with 0.1ml phosphate buffer solution and then centrifuged, repeated 3 times, the supernatant was combined to measure the residual protein amount, and the optimal adsorption t...

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Abstract

The invention discloses a preparation method of artificial liposome, belongs to the field of pharmacy, and relates to a preparation method of protein medicament liposome. In the preparation method, nano spherical grain yolk lecithin zirconium or soya bean lecithin zirconium is used as a carrier, and balsam pear protein MAP30, fleece-flower root protein GAP31, lipase CRL or bovine serum albumin (BSA) is used as a wrapping protein. The preparation method comprises the steps of: adding phosphate buffer solution to the carrier material, stirring under the conditions of the temperature being 4 DEG C and rotation rate being 3500rpm to obtain a colorless and transparent dispersion, dissolving the wrapping protein in a phosphate buffer solution, then slowly dropwise adding to the dispersion, quickly stirring in the low-temperature condition to form stable liposome, centrifugally separating out protein, repeatedly washing, centrifuging, and freeze-drying at low temperature to obtain a finished product. The method has the characteristics of simple process, mild condition, high repeatability, high protein adsorption capacity and stable liposome structure.

Description

technical field [0001] The invention belongs to the field of pharmacy, and relates to a preparation method of protein medicine liposome, in particular to a preparation method of artificial liposome. Background technique [0002] Due to the unique physical, physiological and pharmacological characteristics of liposomes, such as the ability to integrate hydrophilic and hydrophobic drugs, good biocompatibility, low toxicity, no immune system response, and targeted buffering of active drugs at the site of action Therefore, as a therapeutic or diagnostic active agent, the research of liposome carrier has been widely concerned. Since the discovery of liposomes, the size of its products can be effectively controlled from micrometers to nanometers; part of the carrier system has been functionally integrated with polypeptides, proteins and antibodies after surface modification. At present, the FDA has approved more than 25 liposome dosage forms such as amikacin / HSPC / CH / DSPG, muramyl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/127A61K47/24A61K38/00A61K38/38A61K38/43A61K49/00A61P35/00A61P31/18
Inventor 乔元彪
Owner LULIANG UNIV
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