JPS63119848A - Apparatus for producing liposome - Google Patents

Abstract

PURPOSE:To remove a surplus oily matter and an incomplete liposome by providing the refining part of a liposome suspension, the feeding mechanism for feeding the liposome suspension to the refining part and the controlling mechanism for controlling the temp. of a reactor to the optimum temp. to the titled apparatus. CONSTITUTION:The apparatus body has a cylindrical shape and is connected with major equipments of 1-5 after sterilizing them. In the major equipments 1-5, 4 is the reactor, 2, 3 are hollow chambers (the refining parts) provided an ultrafiltration and an auxiliary filter and 5 is a pressure type syringe (the feeding mechanism). And the liposome suspension is produced by accommodating the liposome preparation in the reactor 4 the temp. of which is controlled, followed by agitating while bubbling it with inert gas. And the obtd. liposome suspension is refined by passing it to the refining parts 2, 3 composed of the ultrafiltration and the auxiliary filter.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to a production device for safely and inexpensively mass-producing ribosomal gold.

Background] In recent years, technology that utilizes ribosomes, which are closed vesicles made of double-layered lipid membranes, has gained importance in a wide range of fields including medicine, pharmacy, immunology, and biology. Much knowledge is being accumulated toward practical application.

In other words, due to its quasi-cellular properties, ribosomes not only serve as a model for biological membranes and are the subject of fundamental biophysical chemistry research, but also can contain various substances within the aqueous layer or membrane structure inside the vesicle. Friends, for example, each Tsukuda contrast agent? ¥? As a carrier that efficiently retains and promotes therapeutic drugs or physiologically active substances.

It can be used effectively both in and outside the body.

Its groundbreaking clinical application is expected.

However, in the past, there has been no apparatus for producing large quantities of ribosomes having medically and biologically favorable properties, 9 for example, only one such as shown in FIG. 2 exists.

In FIG. 2, 11 is a water bath, and 12 is an aqueous phase (containing drug).

13 is a Teflon plug, 14 is an ether phase (containing membrane component lipid), 15 is a syringe, and 16 is a syringe.

In this device, when a lipid-containing ether solution]4 is injected into an aqueous phase (maintained at 55-65°C in a water bath 11) with a syringe 16, the injected ether/I'14 is mixed with water 12
during contact with water and rise to the surface of the aqueous solution as vapor.

Lipids form ribosomes.

1"1 Problems to be Solved by the Invention Not only does the device shown in Figure 2 require the handling of ether, which poses a safety problem for each operation, but the amount of ether solution that can be injected is only a few mJ at most. Therefore, it was not possible to produce a large amount of ribosomes.

During production, strict ether injection systems are required, but since injection relies on manual labor, it is impossible to do it accurately. Furthermore, component substances that are easily denatured in organic solvents cannot be used, and excess ether components and incomplete ribosomes cannot be efficiently removed.

(→Friendly Means for Solving the Problems) The present invention comprises a reaction tank containing a ribosome preparation substance, an injection part for injecting an inert gas and a liquid material into the reaction tank,
a stirring mechanism that stirs the 2-ribosome preparation substance accommodated in the reaction tank; a purification section that purifies the 9-ribosome suspension produced in the reaction tank; and a liquid sending mechanism that sends the ribosome suspension to the purification section.

The reactor is characterized by being equipped with a temperature control mechanism and a metal plate to control the reaction tank to an optimum temperature.

The stirring mechanism is preferably a homogenizer or mixer, the purification section is preferably a hollow chamber equipped with an ultrafiltration filter and an auxiliary filter, and the liquid feeding mechanism is preferably a pressurized syringe.

(E) Function In the present invention, a ribosome suspension is produced by storing a ribosome preparation substance in a temperature-controlled reaction tank and stirring while bubbling all the inert gas. The ribosome suspension is then passed through a purification section consisting of an ultrafiltration filter and an auxiliary filter to prepare the ribosome suspension.

(f) Example Embodiments of the present invention will be described based on the drawings.

FIG. 1 shows a schematic diagram of an apparatus according to the present invention.

The main body of the device has a cylindrical shape and is formed by connecting the main parts 1 to 5 after sterilizing them. 1-4
is the fixed body, and 4 is the reaction tank.

2.3 is the hollow chamber (purification section) in which the ultrafiltration filter and all auxiliary filters are installed (purification section) 1! - Show. The ultrap-filter and auxiliary filter installation chamber 2.3 is provided with connecting portions 82 to B5 to which tubes for separating the ribosome dispersion and/or for inlet/output of inert gas are connected. 5 is a pressurized syringe (liquid feeding mechanism), 6.
7 is a constant temperature water circulation part for temperature control (temperature control mechanism), 8 is a motor for driving a homogenizer, and 9 is a sharp knife of the homogenizer.

The rotating shaft of the homogenizer is hollow, and a tube 10 for separating the ribosome dispersion liquid is inserted into the inert gas and liquid material injection chamber. In addition.

Bl is the tube connection for inert gas inlet/output and liquid transfer, Wl~W4 is the connection to the constant temperature device, 86.87 is for inert gas and liquid material injection or ribosome dispersion. This is the preparative tube connection.

Ribosomes were produced using the apparatus configured as described above at 45° C. under a nitrogen gas atmosphere as follows.

First, 5.0 g of monoolein as a ribosome preparation substance.
Egg yolk lecithin 5- Og w phosphatidylserine 3
．． Og, dicetyl phosphate o, 5g? Cholestero-Iv2. Og+ and α-tocophero-/L'0.5
g was added to the reaction tank (4) and thoroughly stirred with a homogenizer (9) while bubbling N2 gas from B6.
An oily liquid of uniform consistency was obtained.

Next, add 0 to this oily liquid, which had been prepared in advance at 45°C.
, 28M glucone-containing physiological saline solution (PH7.0
), add from 3mj'iB7 and stir at low speed for 10 minutes and at high speed for 5 minutes. In this state, 87 to 200 mJ of glucose-free physiological saline was gradually added and the mixture was stirred at high speed for 5 minutes to obtain a suspension of ribosomes that retained all of the glucose.

This suspension can be collected directly from B6, but if it is passed through a polycarbonate ultrap filter 3.2 in order to remove excess oily components and incomplete ribosome QBI and to make the particle size uniform. , Add the physiological saline from B7 and apply pressure with the movable part '5 while washing, and separate the ribosome suspension from iB4 or B2. It has a granular shape and does not aggregate in physiological saline.
It had a high glucose retention rate of 0.5%.

In addition, in the above explanation, a pressurized syringe was explained as the liquid feeding mechanism, but the present invention is not limited to this.9
For example, ribosomes may be washed by connecting a suction device to B1 and adding a washing liquid from B7.

(Toff effect) According to the present invention, excess oily components or incomplete ribosomes and micelles after ribosome production can be easily removed.

It is possible to perform the operations from preparation of ribosomes to fractionation of a dispersion liquid continuously in an aseptic manner.

[Brief explanation of the drawing]

FIG. 1 is a schematic diagram of a ribosome manufacturing device according to the present invention,
FIG. 2 shows a conventional device. 2.3: Ultrafiltration filter and auxiliary filter installation chamber 4: Reaction tank 5: 21 IO pressure syringe 6.7: Temperature control mechanism

Claims (1)

[Claims] 1. A reaction tank containing a ribosome-preparing substance, an injection section for injecting an inert gas and a liquid material into the reaction tank, and a stirring mechanism for stirring the ribosome-preparing substance contained in the reaction tank. and a purification section for purifying the ribosome suspension produced in the reaction tank, a liquid feeding mechanism for sending the ribosome suspension to the purification section, and a temperature control mechanism for controlling the reaction tank at an optimum temperature. A ribosome manufacturing device characterized by: 2. The ribosome manufacturing device according to claim 1, wherein the stirring mechanism comprises a homogenizer or a mixer. 3. The ribosome manufacturing device according to claim 1, wherein the purification section comprises a hollow chamber in which an ultrafiltration filter and an auxiliary filter are installed. 4. The ribosome manufacturing device according to claim 1, wherein the liquid feeding mechanism is a pressurized syringe.