JPS63283748A - Myoglobin adsorption material - Google Patents

Abstract

PURPOSE:To selectively reduce myoglobin in the blood and minimize the removal of useful substances such as albumin, immune globulin or the like by forming a myoglobin adsorption material by water insoluble porous material having a contact angle of 20 deg. or more. CONSTITUTION:A myoglobin adsorption material is formed by a porous material, solid in the water and having a contact angle of 20 deg. or more on between air bubble and solid material surface in the water. The absorption material is enough to constituted by partial to hydrophobic material at least, and an organic high-molecular compound such as polypropylene, polystyrene or the like is preferable. The adequate shape of the water insoluble porous material is globular or granular, and the adequate average grain diameter is 10-2,500mum. Also, the adequate removal critical molecular weight (protein) is in the range of 2X10<4>-2X10<7>, and the adequate size distribution of pores in the porous material is in the range of 50-2,000Angstrom .

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a myoglobin adsorbent that selectively adsorbs myoglobin, which is thought to be closely related to certain types of acute renal failure.

Myonephropatic m is a complication of acute arterial occlusion syndrome, a so-called crush syndrome in which intramuscular myoglobin leaks out due to extensive muscle crush and causes kidney damage.
etabolic syndrome (MNMS)
Many diseases and injuries, such as electric shock, diabetic coma, and sunstroke, can increase myoglobin in the blood, causing damage to the kidneys.
It is known that it can cause kidney failure. Therefore, there is a need for a technology that selectively removes myoglobin from the blood and prevents kidney damage.

(Prior Art) Existing technologies that can be used for the above-mentioned method include (1) plasma exchange and (2) plasma filtration.

Plasma exchange uses blood containing myoglobin.

In this method, plasma, which is a liquid component, is separated by filtration or centrifugation techniques, the plasma containing 2-myoglobin is discarded, and frozen fresh plasma or albumin solution is replenished. However, with this method, useful components in the blood such as albumin, immunoglobulin, blood coagulation system proteins, complement components, hormones, etc. are discarded together with myoglobin, and the replacement fluids such as frozen fresh plasma and albumin solution are Since it is derived from a living body, it has various problems, including the problem of easy infection with infectious diseases, the problem of difficulty in obtaining the raw material plasma, and the problem of high cost. In addition, plasma filtration filters and discards all molecules smaller than myoglobin from the blood.
This is a method of replenishing the electrolyte. This method wastes less material than plasmapheresis. However, because the pore size distribution of blood filtration Di is not constant, it is inevitable that proteins larger than myoglobin will be lost and hypoproteinemia will occur. The drawback was that it could be thrown away.

(Problems to be Solved by the Invention) As described above, with the conventional techniques, the removal of useful components in the blood was unavoidable, and it was impossible to selectively remove myoglobin in the blood.

The purpose of the present invention is to provide a technology for selectively removing myoglobin from blood, and to provide an adsorbent that can selectively adsorb and remove myoglobin while reducing the removal of useful substances such as albumin and immunoglobulin. It is about providing.

(Means for Solving the Problems) As a result of intensive research in line with the above-mentioned purpose, the present inventors have found that myoglobin can be increased to a surprising extent by using a water-insoluble porous body made of a hydrophobic material. The present invention was completed by discovering that high adsorption ability can be exhibited only when the degree of hydrophobicity of the material is 20 degrees or more in terms of contact angle.

That is, the gist of the present invention is a myoglobin adsorbent F characterized by being made of a water-insoluble porous material having a contact angle of 20 degrees or more.

The substance to be adsorbed in the present invention is myoglobin, and to explain it in more detail, it is a monomer with a molecular weight of about 1.7×10', and like hemoglobin, it is a protein that reversibly binds oxygen.

The "contact angle" referred to in the present invention is the contact angle of air bubbles on a solid surface in water, and is the contact angle of air bubbles on a solid surface in water.
J, Co11oid Interface Sci
, 40, 219-222 (1972) and J.D.
, Andrade, J., Polym, Sci.
Polym, Symp.

66.313-336 (1979). In addition, samples were prepared in sheet-like, film-like, or other molded products, and the contact angle was measured at least 10 times at a temperature of 25° C., and the average value was taken as the value of the contact angle of the material.

The material used for the water-insoluble porous body of the present invention has a contact angle of 20
It needs to be above 100%. If the contact angle is smaller than this, the surface of the water-insoluble porous material will be too woody and its ability to adsorb myoglobin will be extremely reduced, resulting in poor practical ability as an adsorbent for blood myoglobin. In addition, if the contact angle is too large, that is, if the degree of hydrophobicity is too high, although the myoglobin adsorption ability is high, non-selective adsorption tends to increase slightly. Therefore, the preferable contact angle range is from 20 degrees to 110 degrees. degree, and the more preferable range is 25 degrees.
The range is from 100 degrees to 100 degrees, more preferably from 30 degrees to 90 degrees.

The water-insoluble porous material having a contact angle of 20 degrees or more in the present invention is a porous material made of a material that is solid in water and has a contact angle of 20 degrees or more as measured by the method described above. In other words, as long as the porous material is made of a material that is biased toward hydrophobicity, it does not matter whether it is an inorganic compound or an organic compound. High molecular compounds are preferred. Such examples include polypropylene, polystyrene, polymethacrylate esters, polyacrylate esters,
Examples include polymers and copolymers of vinyl compounds such as polyacrylic acid and polyvinyl alcohol, polyamide compounds such as nylon 6.66, and polyester compounds such as polyethylene terephthalate. The material of the porous body used in the present invention may have a contact angle of 20 degrees or more, and is not limited to the above examples.

Among the examples, polymers and copolymers of vinyl compounds are more preferably used because of ease of polymerization and ease of pore adjustment. Such examples include styrene, P
- Polymers of styrenic compounds such as methylstyrene and P-ethylstyrene, methyl (meth)acrylate, ethyl (
Polymers of (meth)acrylic acid ester compounds such as meth)acrylate and propyl(meth)acrylate, and combinations of the above compounds with vinyl compounds such as divinylbenzene, methacrylonitrile, vinylpyrrolidone, ethylene glycol, and methyl acrylate. Examples include polymers.

Among these, methyl (meth)acrylate, ethyl (meth)acrylate, propyl (meth)acrylate, etc.
Polymers of meth)acrylic acid ester compounds and copolymers with the above-mentioned vinyl compounds are more preferably used.

Furthermore, the material used for the water-insoluble porous body of the present invention is a polymer of %(meth)acrylic acid ester compound and a copolymer with the vinyl compound containing methyl methacrylate in an amount of 70% by weight or more. , gives more favorable results.

As for the shape of the water-insoluble porous body, any of the shapes such as spherical, granular, filamentous, hollow fiber, and flat membrane shapes can be effectively used, but spherical or granular shapes are particularly preferably used from the viewpoint of body fluid circulation during body fluid circulation. Spherical or granular average particle size is 10-25
00μm is easy to use, but 25μm to 300μm
A range of m is preferably used.

The water-insoluble porous material needs to be porous from the viewpoint of having a large adsorption surface area for myoglobin and achieving a practical adsorption capacity. Porous body exclusion limit molecule ff1
Since the molecular weight of myoglobin (protein) is 1.7X10', it is necessary for the molecular weight to be greater than this value, which is 2X10'.
A range of ' to 2X10' is preferred. More preferred is 2x 10r' to 2x 10'. The pore distribution of the porous body can be obtained from the mercury pressure blood line obtained by the mercury intrusion method (for example, Catalyst Engineering Course-4, Catalyst Measurement Method, edited by the Catalyst Society, Jijinshokan, pp. 69-73). 5O in pore diameter
It is preferable that there are many pores of A or higher, from 50λ to 2000
It is more preferable that the pores are concentrated in the range of λ. More preferably, the range is from 60A to 1000 people, and 65
A range of A to 400 A is most desirable.

The method for producing a water-insoluble porous body will be exemplified below.
The present invention is not limited to this example.

The method for producing the water-insoluble porous material of the present invention is generally known methods such as bulk polymerization, solution polymerization, suspension polymerization, and emulsion polymerization. and a solvent that dissolves the monomer, and are carried out in the respective polymer production methods.

For example, in the case of a styrene-divinylbenzene copolymer, by stirring in the coexistence of styrene, ethylbenzene, divinylbenzene, toluene, octatool, and AIBN (azobisisobutyronitrile), a spherical diameter of about 50 to 1000 μm can be obtained. Porous particles can be made. Particles with various particle sizes and pore sizes can also be produced by radical polymerization in a suspension polymerization system.

The water-insoluble porous body of the present invention can be subjected to surface treatment to inhibit the adhesion of platelets in order to improve its affinity with blood. This can be achieved, for example, by providing a coating layer of a material with low platelet adhesion on the surface of the water-insoluble porous material that comes into contact with blood cells.

Examples of low platelet adhesion materials include polymeric materials having hydroxyl groups such as droxyethyl methacrylate and hydroxyneethyl acrylate, and QLff1 bodies having basic nitrogen-containing functional groups such as vinylamine and dimethylaminoethyl (meth)acrylate. Blocks such as copolymers with polymerizable monomers that do not have basic nitrogen-containing functional groups, polymeric materials that have negatively charged functional groups such as sulfonic acid groups and carboxylic acid groups, segmented polyurethanes, and segmented polyesters. Copolymer, with polyethylene oxide chains,
Examples include graft copolymers such as copolymers of JIT and other polymerizable monomers.

The myoglobin adsorbent of the present invention is generally used by being filled and held in a container equipped with an inlet and outlet for body fluids. In the accompanying drawings, l indicates an example of an adsorption device filled with the myoglobin adsorbent of the present invention, which has a body fluid inlet 5 at one end opening of a cylinder 2 via a backing 4 with a filter 3 stretched inside. Screw the cap 6 and
A cap 8 having a lower body fluid outlet is screwed into the opening at the other end of the cylinder 2 via a backing 4' with a filter 3' stretched inside to form a container.
The adsorbent layer 9 is formed by filling and holding an adsorbent in the gap .

The adsorbent layer 9 may be filled with the myoglobin adsorbent of the present invention alone, mixed with other adsorbents, or filled in 61 layers. The volume of the adsorbent layer 9 is, when used for extracorporeal circulation,
Approximately 50 to 500 mft is appropriate. When the device of the present invention is used for extracorporeal circulation, there are two methods as follows: One method is to separate blood taken from the body into plasma components and blood cell components using a centrifuge or membrane-type plasma separator SI.The plasma components are passed through the device, purified, and then separated into blood cells. One method is to return the blood to the body together with its components, and the other method is to purify the blood taken from the body by passing it directly through a nuclear device.

In addition, regarding the passage speed of blood or plasma, since the adsorption efficiency of the adsorbent is very high, the particle size of the adsorbent can be made coarser, and the degree of packing can be lowered, so the shape of the adsorbent layer can be changed. High passing speeds can be achieved regardless of the Therefore, a large amount of body fluid can be treated.

The method for passing body fluids may be either continuous or intermittently depending on clinical needs or equipment conditions.

(Effects of the Invention) As described above, the myoglobin adsorbent of the present invention selectively adsorbs and removes myoglobin from body fluids at a high rate, and the adsorption device using the adsorbent is extremely compact. It is convenient and safe for soldiers.

The present invention is applicable to general methods of purifying and regenerating body fluids such as blood and plasma, and is effective for safe and reliable treatment of diseases associated with increased myoglobin concentration in the blood.

Furthermore, the adsorbent of the present invention can be used not only as a treatment device by filling it into an apparatus, but also as an affinity adsorbent for separating and purifying myoglobin.

(Example) Hereinafter, embodiments of the present invention will be described in more detail with reference to Examples.

Example! As the water-insoluble porous material, a methyl methacrylate/divinylbenzene copolymer (80:20% by weight, Ambersep, manufactured by Organo) was used.

The contact angle of air bubbles in water of the material used for the water-insoluble porous body was 65 layers and 3 degrees.

The following adsorption experiments were conducted using this water-insoluble porous material as a myoglobin adsorbent.

To 1 ml of water-insoluble porous material, 6 mA of a 120 μg/ml (dissolved in heparinized human plasma) solution of myoglobin (Cubbell, human myoglobin positive control) was added, and the mixture was incubated at 37°C for 1 hour with shaking. After that, the water-insoluble porous material is allowed to settle,
The myoglobin concentration of the supernatant was measured.

As a result of the adsorption experiment, the myoglobin concentration after adsorption was 9.4 μ
g/ml (8% of before adsorption).

Example 2 An experiment was carried out in the same manner as in Example 1, except that a vinyl alcohol/triallyl isocyanurate copolymer was used as the water-insoluble porous material.

The contact angle of air bubbles in water of the vinyl alcohol/trialyl isocyanurate copolymer was 25 layers and 4 degrees.

A vinyl alcohol/triallyl isocyanurate copolymer was obtained as follows. That is, 100g of vinyl acetate
, triallylisocyanurate 64.3g and 2
．． Water in which 2'-azobisisobutyronitrile, polyvinyl alcohol, sodium dihydrogen phosphate dihydrate, and disodium hydrogen phosphate were dissolved was placed in a flask, thoroughly stirred, and suspension polymerized at 75°C for 5 hours. conduct, diameter 350
A granular polymer of ~500 μm was obtained.

As a result of the adsorption experiment, the myoglobin concentration before adsorption was 120 μ
g/m was 11, whereas after adsorption it was 60 μs.
/mjl (50% of before adsorption).

Comparative Example 1 An experiment was carried out in the same manner as in Example 1, except that agarose gel (Sepharose 4B, Pharmacia Japan) was used as the water-insoluble porous material.

The contact angle of air bubbles in water of agarose is 11±6
It cleared up in degrees. As a result of the adsorption experiment, the myoglobin concentration before adsorption was 120μg/mJ! On the other hand, after adsorption, the concentration did not decrease much to 97 μg/ml (81% of before adsorption).

Clinically, it is said that renal failure will not occur if the blood myoglobin concentration of 100 μg/m II is lowered to less than half. The adsorption capacity of this comparative example is not practical because the blood myoglobin concentration cannot be reduced to less than half unless an adsorbent of 111 or more is used.

Example 3 An experiment was carried out in the same manner as in Example 1, except that a styrene/divinylbenzene copolymer (HP-10, Sanyi Kasei Kogyo Co., Ltd.) was used as the water-insoluble porous material.

The contact angle of the air bubbles of the styrene/divinylbenzene copolymer in water was 5 degrees above the flow.

As a result of the adsorption experiment, the myoglobin concentration before adsorption was 120 μ
g/ml, whereas after adsorption it was 8.3 μg.
/mfl (7% before adsorption).

[Brief explanation of the drawing]

The figure is a schematic cross-sectional view showing one example of the form of an adsorption device using the myoglobin adsorption material of the present invention.

Claims (1)

[Claims] A myoglobin adsorbent comprising a water-insoluble porous material having a contact angle of 20 degrees or more.