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Method for producing endoplasmic reticulum

A manufacturing method and vesicle technology, which can be applied to medical preparations without active ingredients, medical preparations containing active ingredients, and pharmaceutical formulas, etc., and can solve the problems of insufficient particle size control and failure to be practical.

Active Publication Date: 2014-03-12
NARA MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Historically, it was disclosed in the late 1960s that phospholipids as amphiphilic molecules self-associate in water to form bimolecular membranes, thereby forming vesicle structures (phospholipid vesicles, liposomes); Attempts to encapsulate hemoglobin in the phospholipid vesicles were not put into practical use due to difficult modulation such as particle size control and insufficient means to inhibit aggregation caused by plasma protein interactions (Non-Patent Document 2: Djordjevich L&Miller IF. Fed Proc.1977;36:567)

Method used

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  • Method for producing endoplasmic reticulum
  • Method for producing endoplasmic reticulum
  • Method for producing endoplasmic reticulum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] As complex lipids, a total of 10 g of 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC), cholesterol, 1,5-O-dicetyl-N-succinyl -glutamic acid (DHSG), and 1,2-distearoyl-sn-glycerol-3-phosphatidylethanolamine-N-polyethylene glycol 5000 (DSPE-PEG5000, the molecular weight of the PEG chain is 5000) according to mole The ratio was 5 / 4 / 0.9 / 0.03, dissolved in 1 dL of tert-butanol, and poured into a 0.5 L eggplant-type flask. Next, freezing was carried out in a dry ice / methanol mixed cooling medium, and it was carried out for 24 hours in a freeze-drying apparatus (FD-1000 manufactured by Tokyo Rika Co., Ltd.) to obtain a composite lipid powder. 10 g of this powder was put into a THINKEY CORPORATION cylindrical container made of Teflon (outer diameter: 76 mm, height about 93 mm, inner wall with unevenness, clover shape when viewed from above the container), and high-purity human hemoglobin was added Solution (carbon monoxide binding-HbCO body, 45g / dL, 0.4dL, pH7.4). The...

Embodiment 2

[0059] Using the same lipid species as in Example 1, DPPC / cholesterol / DHSG / DSPE-PEG5000 was dissolved in 1 dL of tert-butanol so that the molar ratio was 5 / 4 / 0.9 / 0.03 and the total lipid weight was 10 g. , added to an eggplant-shaped flask. Next, it was freeze-dried with liquid nitrogen to obtain complex lipid powder. Next, this complex lipid was put into the same Teflon cylindrical container as that used in Example 1, and 0.5 dL of ultrapure water was added thereto. The weight of the negatively charged lipid DHSG was 1.13 g, corresponding to 1.48 m mole, and 0.074 mL of 1N-NaOH solution was added in order to neutralize it with an equivalent amount of NaOH. Then, cover the inner cover and seal it, use a kneading device (rotation and revolution agitator, manufactured by THINKEY CORPORATION, ARE-310) to knead for 5 minutes (revolution 2000 rotations, rotation 800 rotations), after cooling for 5 minutes, carry out again Kneading was performed for 5 minutes, and the treatment wa...

Embodiment 3

[0061] Manufactured by Nippon Fine Chemical Co., Ltd. Dissolve DPPC / cholesterol / DHSG / DSPE-PEG5000 in an organic solvent so that the molar ratio becomes 5 / 4 / 0.9 / 0.03, and use the CRUX method to make the organic solvent Complex lipid powder obtained by rapid evaporation. For 10 g of this complex lipid powder, in the same manner as in Example 1, 0.4 dL of thick hemoglobin solution was added, and a kneading device (autorotation revolution agitator, manufactured by THINKEY CORPORATION, ARE-310, revolution speed 1000 revolutions, rotation speed 400 revolutions) the operation of kneading for 6 minutes and cooling for 3 minutes was repeated 30 times. The temperature rise of the paste stayed around 30°C. All kneading was carried out for 180 minutes. At this stage, the viscosity of the obtained paste was measured at 25°C with a rheometer (MCR-300, manufactured by Anton Paar Co.), and it was about 2200cP (shear rate 1000s -1 , 23℃), about 10000cP (shear speed 100s -1 , 23℃), about 52...

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Abstract

A method for producing an endoplasmic reticulum in which a functioning substance is included into a lipid endoplasmic reticulum having lipids as the main component. This method involves a step (a) for storing the functioning substance, lipids and water into a tubular container, and a step (b) for producing the endoplasmic reticulum in which the functioning substance is included into a lipid endoplasmic reticulum having lipids as the main component by kneading the contents stored in the tubular container as a consequence of rotating the tubular container about the central axis thereof and making the tubular container orbit about a predetermined orbital axis. DRAWING: FIG.1 AA Intersection point BB Rotation axis CC Rotation DD Rotation angle velocity EE Orbital axis FF Rotation radius (r) GG Orbit HH Orbital axis velocity II Orbit radius (r)

Description

technical field [0001] The present invention relates to a method for producing a phospholipid vesicle (liposome) preparation. In particular, it relates to a method for producing a dense dispersion of an artificial oxygen carrier (hemoglobin vesicle) used instead of blood transfusion in the medical field more efficiently than conventional techniques. Background technique [0002] The current blood donation-blood transfusion system has been established as an indispensable technique in medicine. However, along with various problems (infection, shelf life, decrease in the number of blood donations, etc.), the search for transfusions instead of blood transfusions has become an urgent issue. In particular, it is sought to realize a substitute for the red blood cell, which holds the oxygen-carrying function. Hemoglobin (Hb) contained in a high concentration in erythrocytes is an oxygen-binding protein, and therefore the development of artificial erythrocytes using this hemoglobin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/127A61K38/16A61K47/18A61K47/24A61K47/28A61K47/44
CPCA61K38/42A61K9/1271A61K9/0026A61K9/1277A61J3/00
Inventor 酒井宏水
Owner NARA MEDICAL UNIVERSITY