Lactoferrin modified patchouli alcohol liposome as well as preparation method and application thereof

A technology of lactoferrin and patchouli alcohol, applied in the field of medicine, can solve the problems of difficult to exert curative effect, poor drug absorption, low bioavailability, etc. Effect

Active Publication Date: 2021-10-26
SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First of all, most insoluble drugs have poor drug absorption in the body and low bioavailability, so it is difficult to exert curative effect

Method used

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  • Lactoferrin modified patchouli alcohol liposome as well as preparation method and application thereof
  • Lactoferrin modified patchouli alcohol liposome as well as preparation method and application thereof
  • Lactoferrin modified patchouli alcohol liposome as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1: the preparation of the liposome of lactoferrin modification

[0063] Weigh egg yolk lecithin, cholesterol, distearoylphosphatidylethanolamine-polyethylene glycol 2000, distearoylphosphatidylethanolamine-N-hydroxysuccinimide-polyethylene glycol 2000 (30:6:6 :1,w / w) (both purchased from Shanghai Aiweite Pharmaceutical Technology Co., Ltd.) and patchouli alcohol (Chengdu Master Biological Technology Co., Ltd.), dissolved in chloroform. The resulting solution was evaporated to dryness with chloroform under reduced pressure at 37°C to form a thin film. Phosphate buffer solution was added to the film for hydration to form lipid water dispersion. Ultrasonic cell disruptor (JY92-IIN, Ningbo Xinzhi Biotechnology Co., Ltd.) was used to sonicate for 5 min at 5% power. Afterwards, use a membrane extruder to repeatedly extrude the liposomes until the liposomes pass through a membrane with a particle size of 200 nm to form liposomes with uniform particle sizes. A Sep...

Embodiment 2

[0064] Example 2: Characterization of lactoferrin-modified liposomes

[0065] (1) Reaction of lactoferrin with N-hydroxysuccinimide ester

[0066] Weigh 1 mg of lactoferrin, prepare a 1 mg / mL solution with ultrapure water, and use the same method to prepare a distearoylphosphatidylethanolamine-N-hydroxysuccinimide-polyethylene glycol 2000 solution. The lactoferrin solution was mixed with the distearoylphosphatidylethanolamine-N-hydroxysuccinimide-polyethylene glycol 2000 solution, placed on a shaker at 4°C, and incubated for 12-16 hours. Take the reacted solution, place it in a dialysis bag with a molecular weight of 14K, and take it out after dialysis with magnetic stirring for 24 hours. At this time, the reaction product distearoylphosphatidylethanolamine-lactoferrin-polyethylene glycol 2000 is obtained. Mix lactoferrin solution, distearoylphosphatidylethanolamine-lactoferrin-polyethylene glycol 2000 solution with sinapinic acid matrix (Shanghai Yuanye Biotechnology Co., Lt...

Embodiment 3

[0081] Example 3: Cell Safety Evaluation

[0082] Bone marrow-derived macrophages (extracted from Balb / c mice) were mixed at 1×10 per well 4 Cells were seeded in 96-well plates, 100 μL per well, and induced to differentiate into M1 macrophages. After the cell differentiation was completed, they were divided into three groups: free patchouli alcohol, liposomes prepared in Example 1, and lactoferrin-modified liposomes prepared in Example 1, and each group was administered according to a certain concentration gradient. After administration, the culture was continued for 24 hours, and then 20 μL of thiazolium blue (MTT) (Sigma-Aldrich, USA) was added to each well, and the incubation was continued for 4 hours, after which the supernatant was aspirated to leave purple crystals. Add 200 μL dimethyl sulfoxide to each well, place on a shaker at room temperature until the purple crystals are completely dissolved, use a microplate reader (Multiskan, ThermoFisher, USA) to detect the OD v...

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Abstract

The invention provides a lactoferrin modified liposome. The lactoferrin modified liposome is prepared from the following raw materials: lactoferrin, egg yolk lecithin, cholesterol, distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000, distearoyl phosphatidyl ethanolamine-N-hydroxysuccinimide-polyethylene glycol 2000 and patchouli alcohol, wherein the ratio of the egg yolk lecithin to the cholesterol to the distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000 to the distearoyl phosphatidyl ethanolamine-N-hydroxysuccinimide-polyethylene glycol 2000 is (29-32): (5-7): (5-7): (0.5-1.5) (w / w); the patchouli alcohol is wrapped in the liposome; and amino groups contained in the lactoferrin react with N-hydroxy succinimide in the distearoyl phosphatidyl ethanolamine-N-hydroxy succinimide-polyethylene glycol 2000, so that the amino groups contained in the lactoferrin are connected to the surface of the liposome.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a lactoferrin-modified patchouli alcohol liposome, a preparation method thereof and an application in inflammatory bowel disease. Background technique [0002] Inflammatory bowel disease is a chronic relapsing idiopathic gastrointestinal inflammation that can lead to long-term and even irreversible damage to the structure and function of the gastrointestinal tract, including the two manifestations of Crohn's disease and ulcerative colitis. Crohn's disease is common at the end of the small intestine and the beginning of the colon, and may involve any part of the gastrointestinal tract in different forms; ulcerative colitis begins in the rectum and can extend up the entire colon. The etiology and pathogenesis of inflammatory bowel disease are still unclear, and it may be related to genetic susceptibility, imbalance of intestinal microbial homeostasis, impaired intestin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/127A61K47/42A61K47/24A61K31/045A61P1/00A61P1/04A61P29/00
CPCA61K9/1271A61K47/42A61K47/24A61K31/045A61P1/00A61P1/04A61P29/00
Inventor 黄永焯王冰赵宇戈
Owner SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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