JP3144897B2 - Filter unit for producing suspended lipid particles and method for producing suspended lipid particles - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a filter unit suitable for producing suspended lipid particles typified by emulsions and liposomes, and to a method for producing finely divided suspended lipid particles within a predetermined range.

[0002]

2. Description of the Related Art For example, lipid particle suspensions such as emulsions in which water and oil are emulsified with amphipathic lipids and liposomes having an aqueous phase therein and closed vesicles by lipid bilayer membranes are known. 2. Description of the Related Art There has been much interest in practical use in pharmacological applications to anticancer drugs, anti-AIDS drugs, etc., food industry applications, and cosmetics industry applications. It is desired that this type of suspended lipid particles (lipid suspension particles) have a limited average particle size and a limited particle size distribution.

In response to such demands, (a) a production method in which the suspended lipid particles are miniaturized using mechanical shearing force such as a high-pressure homogenizer, or (b) a filter / filter is used. For example, there is known a manufacturing method in which a pressure of 1 to 7 kgf / cm ² is applied to force the liquid to pass through to make finer.

[0004]

However, the above-mentioned method and means for producing fine lipids have the following disadvantages in practical use. That is, in the case of the method or means (a), a part of a high molecular drug such as a peptide or a protein is cut or denatured due to the strong shearing force, and the high heat applied partially causes There is a problem that precise control of the particle size is practically difficult because not only does a heat-sensitive drug decompose, but also it relies on mechanical emulsification (lipid refinement). On the other hand, in the case of the method or means (b), the type and configuration of a filter, for example, a membrane filter, and the combination with a porous filter support that mechanically supports and supports the selected membrane filter are considered. Therefore, the fact is that the suspension lipid particles cannot be efficiently refined or mass-produced. To further mention this point, in the case of a membrane filter in which required micropores are provided in a resin film by selective etching, the effective area is reduced and the efficiency of the micronization is inferior. The required particle size cannot be achieved with a membrane filter in which the holes are provided in a sponge shape. In other words, in the case of (b), the required miniaturization of the suspended lipid particles can not be achieved efficiently and in mass production.
Only liters of suspension can be processed, resulting in production costs.
The fact is that the strike is high. Suspension
Increase the supply pressure or increase the area of the membrane filter.
Mass production by increasing the amount of suspension that can be processed at one time
It may be possible to improve the performance, but the conventional filter support
Because the body is made of porous plastic,
Increase pressure or increase area of membrane filter
Deformation may cause damage to the membrane filter.
Problem. In addition, although the filter support is so-called porous, the aperture ratio is limited,
The flow of the suspended lipid particles (lipid suspension particles) that have been miniaturized by pressurizing and passing through the micropores of the membrane filter is inhibited,
As a result, the required miniaturization cannot be efficiently and mass-produced. The problem due to this filter support is:
For example, it is conceivable to increase the diameter of the through hole of the filter support and to set a large number of the through holes, but on the other hand, there is a problem that the function of mechanically reinforcing and supporting the membrane filter cannot be achieved.

[0005] The present invention has been made in view of such circumstances, and a single processing operation can be performed in units of liters of suspension.
Stepped Filler for Producing Suspended Lipid Particles Having High Mass Production Capability and Effectively Achieving Refinement of Lipid with Required Limited Average Particle Size and Limited Particle Size Distribution
And an object thereof is to provide a method for producing terpolymers units and suspended lipid particles.

[0006]

According to the present invention, a liter unit is provided.
Lipid particles for treating a suspension of lipid particles to be treated in different amounts
The filter unit for the production of particles
The liquid inlet and the treated lipid particle suspension are drained.
At least 5 kgf /
a housing capable of withstanding the supply pressure of cm ^2, one main side is open to the large diameter, the other principal surface side have a stepped through-hole opened to the small diameter,
Arranged in the housing with the large-diameter opening side facing the supply port side
And the supply port side of the suspension of the lipid particles to be treated is
A multi-hole support plate for partitioning the processed lipid particle suspension from the take-out port side, and provision of fine holes penetrating in the thickness direction disposed on the large-diameter opening surface of the porous support plate. And a membrane filter main body through which the suspension of the lipid particles passes.

[0007] The method for producing suspended lipid particles according to the present invention is directed to a filter unit for producing suspended lipid particles according to claim 1.
A suspension containing lipid particles in an amount of liter unit is supplied from the supply port of the knit at a supply pressure of at least 5 kgf / cm ² , and the suspension is supplied to the fine pores and the porous support plate penetrating through the membrane filter body. By passing through the stepped through-holes, the lipid particles contained in the suspension are miniaturized.

[0008] The present invention provides a filter unit comprising a combination of a membrane filter body having through-holes formed therein by, for example, irradiation of charged particles and alkali etching, and a porous support plate having stepped through-holes. Based on the experiment that lipid particles having a required limited average particle size and a limited particle size distribution can be easily obtained by supplying a suspension of lipid particles under pressure using this filter unit. It was made.

Here, the aperture ratio of the membrane filter body due to the penetrating micropores is, for example, about 0.1 to 18%, and the large-diameter aperture and the small-diameter aperture of the porous support plate having the stepped through-holes. Select and set the small aperture diameter in the range of 0.01 to 0.5 when the large aperture diameter is set to 1.
I do. In this range, the suspension of lipid particles in liter units
The effective fineness of the membrane filter body
Damage to the membrane filter body while securing the hole area
Can be reliably supported without the need for
It is not limited to this range. The aperture ratio of the large-diameter aperture of the porous support plate having a stepped through hole is 30 to 90%,
Preferably, it is about 60 to 70%. Further in the above,
The supply pressure applied to the suspension of lipid particles is not particularly limited, but the lower limit is 5 kgf / cm ² . The present invention
Treatment of liter-unit suspension in
As a result, more than one liter of suspension can be
It means that the liquid can be processed, and it was processed several times little by little
It also means that the total processing amount will be 1 liter or more
Not.

The membrane filter body which can be used in the present invention is made of, for example, a polycarbonate film, a polyester film, a polysulfone film, a polyethylene film, a polypropylene film or the like, and is irradiated with charged particles and alkali-etched in the thickness direction as described above. Examples include a member provided with a penetrating fine hole, and a member provided with a fine hole penetrating in the thickness direction by other means. And the diameter of the through micropore is 0.010
1414 μm, preferably 0.015-3 μm, more preferably 0.1-1 μm, and it is desirable that these micropores are provided in a distribution state as uniform as possible.

[0011]

According to the filter unit of the present invention, the porous support plate that contributes to mechanical reinforcement and support of the membrane filter body has a large opening on one main surface and a small opening on the other main surface. It has a stepped through hole. And since the membrane filter main body is arranged on one main surface side of the porous support plate having a large diameter opening, the non-contact area surface is also relatively large, and the lipid particles can pass through the membrane filter main body. Through micropores are sufficiently ensured (effective aperture ratio is improved). On the other hand, the other main surface side of the porous support plate having the stepped through-holes, which is opened to a small diameter, and, if necessary, on this surface side, combined with a support for supporting the porous support plate, In addition, it provides a stable support function. Therefore, when a filter unit of the structure, it is possible to supply the membrane filter body of a suspension of lipid containing particles 5 kgf / cm ² or more high-pressure, the through-microporous membrane filter body and pressurizing and passed through efficient miniaturization of the lipid particles of the suspension is achieved Rutotomoni, one operation
Makes it possible to process suspensions in liters
Not only improves productivity, but also has a very high recovery rate.
It becomes .

In particular, as a membrane filter body,
For example, when using a membrane filter body provided with through-pores with a constant fine pore diameter by irradiation of charged particles and alkali etching, when the lipid particles in the suspension are pressurized and passed, the suspended The particle size of lipids (particles) will easily take a constant average particle size and a limited particle size distribution.

[0013]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS An embodiment of the present invention will be described below with reference to FIGS.

Embodiment 1 First, an embodiment of a filter unit according to the present invention will be described.

FIG. 1 is a cross-sectional view showing an example of a main configuration of a filter unit. Reference numeral 1 denotes a pressurizing / supplying port 2 for a suspension-suspended lipid particle, and removal of a finely treated lipid suspension lipid particle. Maximum supply of suspension in the embodiment with mouth 3
4 is a porous support plate having a stepped through hole 4a provided inside the housing 1 and having a structure capable of withstanding a pressure (40 kgf / cm ² ).
Is a membrane filter main body disposed on the upper main surface of the porous support plate 4 made of metal having the stepped through hole 4a, for example, a stainless steel porous support plate 4, and the stepped through hole 4a is open to a large diameter. This is a porous support plate for mechanically reinforcing and supporting the porous support plate 4 having 4a. It is desirable that the support plate 6 for supporting the reinforcement is arranged in practical use, but may be omitted depending on the use conditions.

FIG. 2 shows the structure of a filter unit including the porous support plate 4 having the stepped through-holes 4a and the membrane filter body 5, that is, the stepped through-holes 4a of the porous support plate 4 are opened to a large diameter. It is sectional drawing which expands and shows the state which arrange | positioned the membrane filter main body 5 on the upper main surface and made it a filter unit.

In the configuration of the filter unit,
The porous support plate 4 having the stepped through-holes 4a has an opening on one main surface side having a large diameter of, for example, about 0.85 mm, and an opening on the other main surface side, for example.
An opening ratio of about 65% on the side with a large opening having a stepped through hole 4a with a small opening of about 0.5 mm, for example, thickness
It is a stainless steel porous support plate of about 0.5 to 0.7 mm,
The membrane filter body 5 has a thickness of, for example, 6 mm.
After irradiating charged particles to a polycarbonate film of about 20 .mu.m, the film is selectively etched using an alkaline solution such as NaOH to form through-holes having a diameter of 0.2 .mu.m. The opening ratio of the membrane filter body 5 is determined by the diameter of the through-holes.
In the case of 0.2 μm, it is around 10%. Further, the porous support plate 6 is, for example, 21 mm thick on a 4 mm thick stainless steel plate.
The through holes (chamfered) with a diameter of 4 mm are formed at approximately equal intervals on concentric circles at intervals, but other configurations may be employed.

Embodiment 2 Next, an example of producing (refining) the required suspended lipid particles using an apparatus in which the filter unit having the above-described structure is provided inside the casing 1 will be described. The measurement of the particle size and the particle size distribution of the lipid particles of the liposome suspension in this example was performed using a Nicomp 370 series (trade name) of Particle Sizing System.

First, a liposome having a particle size distribution shown in FIG.
Yolk lecithin / cholesterol / yolk phosphatidylglycerol = 10 mM / 8 mM / 2 mM)), and 2 liters of the suspension to be treated is applied through a pressure / supply port ^{2 at} a pressure of 20 kgf / cm ² . The stepped filter is injected into a device (housing) internally provided in the housing 1, and a filter unit internally and supported in the housing 1 of the device, that is, a stainless steel having a stepped through hole 4 a After finely penetrating the micropores of the membrane filter body 5 (pore diameter 0.2 μm) arranged on the surface where the stepped through-holes 4a of the porous support plate 4 are open to a large diameter, the fine pores are refined. The micronized liposome suspension was recovered from the outlet 3 through the stepped through hole 4a of the support plate 4.

After the above-described operation of the micronization treatment was further repeated once, the particle diameter and the particle diameter distribution of the lipid particles in the liposome suspension were measured. As shown in FIG.
For example, it was possible to obtain a liposome suspension exhibiting a certain average particle size suitable for an anticancer drug, an anti-AIDS drug, and the like, and a particle size distribution limited to this particle size. The amount of lipid recovered from the outlet 3 was 99.2% of the amount injected into the pressurizing / supplying port 2, and almost the entire amount could be recovered.

That is, in the above method for producing a fine lipid of a liposome suspension, the fine pores of the membrane filter main body 5 are not clogged or the membrane filter main body 5 is not damaged. The flow of the suspended lipid particles can be easily and efficiently prevented without any hindrance by the porous support plate 4.
Through the stepped through-hole 4a, it was possible to collect from the outlet 3 of the apparatus main body 1.

In the above embodiment, even if the suspended lipid particles (lipid suspended particles) are changed, the pressure at the time of supplying the suspended lipid particles under pressure is 5 kgf / cm ² , 10 kgf / cm ² , 15 kgf. /
^{cm 2, 20kgf / cm 2,} 25kgf / cm 2, 30kgf / cm 2, 35
kgf / cm ^2, or be modified with 40 kgf / cm ^2, results as in the case of the were obtained. Further, in the configuration of the filter unit, the membrane filter body 5
A similar result was also obtained by adopting a configuration in which both sides were sandwiched between mesh screens and arranged on a porous support plate 4 having stepped through holes 4a.

[0023]

As is clear from the above examples, according to the filter unit for producing suspended lipid particles of the present invention and the method for producing suspended lipid particles using the filter unit for producing suspended lipid particles, Suspended lipid particles having a certain average particle size required for mass production and a limited particle size distribution with respect to this diameter can be easily obtained. For example, pharmacology of anticancer drugs, anti-AIDS drugs, etc. It has many practical advantages in industrial applications, food industry applications, or cosmetic application industries.

[Brief description of the drawings]

FIG. 1 is a sectional view showing an example of a configuration of a main part of a filter unit according to the present invention.

FIG. 2 is an enlarged sectional view showing the configuration and arrangement of a membrane filter main body and a porous support plate 4 having a stepped through hole 4a of the filter unit according to the present invention.

FIG. 3 shows an example of the particle size distribution of lipid particles produced by the method for producing suspended lipid particles according to the present invention, wherein (a) is a particle size distribution diagram of suspended lipid particles before miniaturization, b) is a particle size distribution diagram of the suspended lipid particles after miniaturization.

[Explanation of symbols]

DESCRIPTION OF SYMBOLS 1 ... Housing 2: Pressurization / supply port of suspended lipid particles 3 ...
Port for taking out the microparticle-treated suspended lipid particles 4 Metal porous support plate having stepped through holes 4a Stepped through holes 5 Membrane filter body 6 Support rod

──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Taikazu Hirose 948-1, Okada, Atsugi-shi, Kanagawa Nomura Micro-Sciences Corporation Development Business Headquarters No. 16-13, Daiichi Pharmaceutical Co., Ltd., Central Research Laboratory Formulation Center (72) Inventor Kiyoto Taniuchi 1-16-13 Kita-Kasai, Edogawa-ku, Tokyo Daiichi Pharmaceutical Co., Ltd., Central Research Laboratory Formulation Center (56 References: Japanese Utility Model Application Showa 48-52174 (JP, U) Special Tables Showa 61-502452 (JP, A) (58) Fields investigated (Int. Cl. ⁷ , DB name) B01F 1/00-5/26 B01D 61/00-71/82

Claims (2)

(57) [Claims] 1. A method for supplying a suspension of lipid particles to be treated.
Feed and take-out where the suspension of treated lipid particles is discharged
To a supply pressure of at least 5 kgf / cm ² with a mouth
A housing capable of withstanding, one main side is open to the large diameter, the other principal surface side have a stepped through-hole opened to the small diameter, the large diameter opening side in the housing wherein
A suspension of the lipid particles to be treated, arranged on the supply port side
Supply side and removal of the suspension of lipid particles after the treatment
A porous support plate for partitioning the mouth side, and a membrane filter main body through which a suspension of the lipid particles provided with micropores penetrating in the thickness direction arranged on the large-diameter opening surface of the porous support plate passes. Suspension of lipid particles to be treated in an amount of liter unit, characterized by comprising:
A filter unit for producing suspended lipid particles . 2. The suspension for producing suspended lipid particles according to claim 1,
A liter-unit amount of the suspension containing lipid particles was supplied from the supply port of the filter unit at least 5 kgf / cm ² .
The suspension is supplied at a supply pressure , and the suspension is passed through the fine pores of the membrane filter body and the stepped through-holes of the porous support plate, whereby the lipid particles contained in the suspension are refined. A method for producing suspended lipid particles.