JP2007260216A - Adsorbent carrier and column for extracorporeal circulation - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an adsorbent carrier capable of removing cells in the blood, especially activated white blood cells of granulocyte, monocyte, cancer cells, and excessively-contained cytokine. <P>SOLUTION: An adsorbent carrier manufacturing method is the method for treating the adsorbent carrier, where an iodine content exceeds 1.4 wt.%, with a liquid containing a halogen compound excluding an iodine compound so as to allow the iodine content to be not more than 1.4 wt.%. The adsorbent carrier of the present invention is the one manufactured by the method. An iodine removal treatment is performed with the liquid containing the halogen compound excluding the iodine compound. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a novel adsorption carrier, particularly an adsorption carrier used for medical use.

  In recent years, various blood processing columns have been studied. For example, columns for removing leukocytes and granulocytes (Patent Documents 1 and 2), columns for adsorbing cytokines (Patent Documents 3 and 4), leukocytes and Columns (patent document 5) and the like aimed at simultaneously adsorbing toxins have been developed. These usually have a filter medium or an adsorption carrier for removing and adsorbing a target substance in the column. Various functional groups may be introduced on the surfaces of these adsorption carriers for the purpose of improving biocompatibility or providing selective adsorption ability for specific substances. Examples of the introduction method include immobilization of a functional group by a chemical reaction and introduction by coating. Among such functional groups, a functional group having a charge such as an ammonium group may be introduced for the purpose of adsorbing proteins such as cytokines in particular (Patent Document 6). Cytokines are necessary substances for living organisms, but certain cytokines overproduced due to some disease have negative aspects such as disturbing immunity, and thus overproduced cytokines are efficient. It is desirable that it can be adsorbed or removed, and an efficient method for producing such an adsorbent carrier is required.

On the other hand, there are cases where iodide ions are used as a catalyst for introduction of ammonium groups (Patent Document 6), but these iodide ions are likely to remain on the adsorption carrier as counter ions of ammonium groups. However, focusing on these remaining iodide ions, there is no research on improving the cytokine adsorption rate by the adsorption carrier, and it is also known that there is a correlation between the remaining amount of iodide ions and the cytokine adsorption rate. It was not done.
JP-A-60-193468 JP-A-5-168706 Japanese Patent Laid-Open No. 10-225515 JP-A-12-237585 JP 2002-113097 A Japanese Patent Application Laid-Open No. 2003-11834

  In view of the problems of the prior art, the present invention is an adsorption carrier to which an iodine compound is added, and from the viewpoint of the residual amount of iodine, an adsorption carrier capable of efficiently removing excess cytokines in blood. The issue is to provide.

  The adsorption carrier in the present invention is characterized in that the iodine content after the iodine removal treatment is 1.4 wt% or less for the adsorption carrier containing iodine in an amount exceeding 1.4 wt%. Here, the “adsorption carrier containing iodine in an amount exceeding 1.4 wt%” in the present invention is distinguished from an adsorption carrier whose iodine content is 1.4 wt% or less from the beginning. . Moreover, the content rate in the adsorption carrier of iodine after this iodine removal process means the content rate at the time of use of an adsorption carrier.

  In addition, as a method for producing such an adsorbent carrier, a step of introducing a functional group into the adsorbent carrier using a compound containing iodine (iodine compound) as a reaction reagent, and the adsorbent carrier is treated with a liquid containing a chlorine compound after the reaction. And the step of setting the iodine content to 1.4 wt% or less.

  According to the present invention, it is possible to increase the adsorption removal rate of a specific cytokine by the adsorption carrier by setting the iodine remaining in the adsorption carrier to a predetermined amount or less. In addition, since it is possible to find the necessary and sufficient number of treatments (treatment amount) when treating with a liquid containing a chlorine compound to obtain such an adsorption carrier under a predetermined condition, an efficient method for producing an adsorption carrier is provided. It becomes possible.

  The present invention provides a high-efficiency selection of physiologically active substances such as cytokines that are excessively present in blood even when adding a predetermined amount or more of an iodine compound to the adsorption carrier that is a textile product. Studying a method for producing an adsorbent carrier that can be adsorbed and removed in an effective manner, and reducing the amount of iodine remaining on the adsorbent carrier to 1.4 wt% or less, thereby improving the adsorption performance of cytokines such as interleukin-6 (hereinafter, IL-6). It has succeeded in improving and stabilizing. The cause is not necessarily clear, but the remaining iodide ions are easily oxidized, and iodine can be deposited on the surface of the adsorption carrier. Such precipitation may inhibit the adsorption of substances such as cytokines intended for adsorption removal. Alternatively, since the affinity of the adsorption carrier with the cation differs depending on the type of counter ion, it is possible that the interaction force between the adsorption carrier and the substance intended for adsorption removal may be different.

  Examples of iodine compounds in the present invention include iodine, iodides such as potassium iodide and tetraammonium iodide, and compounds in which iodine is covalently bonded, such as organic iodides, but are not limited thereto. is not.

  There are no particular limitations on the method of adding and using the iodine compound to the adsorption carrier in the present invention, and methods such as reaction raw materials, catalysts, ion exchange, washing treatment, and coating can be used. For example, in the nucleophilic substitution reaction of amines in alkyl halides, when alkyl halides are chlorine compounds, iodides such as potassium iodide may be used as catalysts because of low reactivity. Moreover, the halogen itself of the alkyl halide may be referred to as iodine. Moreover, you may use for another reaction in a manufacturing process, washing | cleaning, disinfection, etc. The remaining form of iodine includes both iodine and iodine ions, and examples thereof include iodine, iodide ions, and triiodide ions. When a cation is contained in the adsorption carrier, it may exist as an iodide ion or a triiodide ion as the counter ion. Further, when they are oxidized, they may be precipitated as iodine on the surface. Further, it may be coated on the surface for imparting antibacterial properties.

  Any measurement method can be used for measuring the residual amount of iodine in the present invention, and for example, elemental analysis, fluorescent X-ray analysis, titration, and the like can be used. However, if iodine remains as an iodine molecule instead of an ion and a drying step is not included before using the adsorption carrier, iodine may be sublimated if a step such as vacuum drying is included only during measurement sample preparation. Yes, it is impossible to measure the exact residual amount of iodine when using the adsorption carrier. For this reason, such an adsorption carrier must not be subjected to a drying step when preparing the sample for measuring the amount of iodine remaining.

  Here, the adsorption carrier used in the present invention must have excellent biocompatibility when used for extracorporeal circulation of blood, and has a basic functional group that does not easily adhere to platelets. Preferably there is. Examples of basic functional groups in the present invention include, but are not limited to, primary amino groups, secondary amino groups, tertiary amino groups, and quaternary ammonium groups. Examples of the quaternary ammonium group include those in which triethylamine, tripropylamine, tributylamine, dimethylhexylamine, dimethyloctylamine, dimethyldecylamine, dimethyllaurylamine and the like are chemically immobilized on an alkyl chain or the like. However, it is not limited to these.

The adsorption carrier used in the present invention is preferably one having a felt shape in that it has a low pressure loss and a large surface area in consideration of allowing a liquid to permeate and adsorb. preferable. The felt in the present invention refers to a fiber sheet formed by mechanically, chemically, or thermally treating fibers and bonding them with an adhesive or the adhesive strength of the fibers themselves. Examples thereof include a punching method and a melt blow method, but are not limited thereto.
In the present invention, an adsorption carrier obtained by adding an iodine compound such that iodine contained in the adsorption carrier exceeds 1.4 wt%, wherein the iodine content is 1.4 wt% or less. In order to introduce various functional groups into the adsorption carrier in the production process, the adsorption carrier is treated with a reaction reagent containing an iodine compound and then the iodine remaining in the adsorption carrier is removed in the production process. , A method of washing with an appropriate washing solution, or a method of treating with a treatment solution containing a halogen compound other than an iodine compound and replacing iodide with a chloride, etc., particularly a treatment containing a halogen compound other than an iodine compound. A method of treating with a liquid is preferred. Among these, considering the affinity with water, the method of treating with a treatment liquid containing a chlorine compound (chloride) is most preferable.
The reaction reagent in the present invention means a substance used in the production process of the adsorption carrier, for example, a raw material used when introducing a functional group, a catalyst, a coating agent, or a substance used as a cleaning agent during washing. However, it is not limited to these. When the functional group immobilization method is immobilization of amine by nucleophilic substitution reaction of amine with alkyl halide, the reaction reagent is preferably an iodine compound having high reactivity with alkyl halide. Alternatively, when the alkyl halide is a chlorine compound, the reaction reagent is preferably an iodide such as potassium iodide as a catalyst.

Examples of the chlorine compound in the present invention include chlorides such as sodium chloride and ammonium chloride, but are not limited thereto. The method of use is not limited. For example, when cations are contained in the adsorption carrier and iodide ions remain as counter ions, iodide ions remain by ion exchange by treatment with a solution containing chloride ions. It can be used to reduce the amount.
There is no limitation in particular in the form of the adsorption carrier in this invention, For example, fiber materials, such as a knitted fabric, a nonwoven fabric, and a felt, can be used. As a material for the adsorption carrier, a known polymer such as polyamide, polyester, polyacrylonitrile polymer, polyethylene, polypropylene, polystyrene, or a derivative thereof can be used. A single yarn of these polymers or a mixed system may be used. For example, if a fiber material is used as the form of the adsorption carrier, a core-sheath type, sea-island type, or side-by-side type composite yarn may be used. In addition, the cross-sectional shape of the fiber may be a circular cross-section or other irregular cross-section. Moreover, the diameter of the cross section of the fiber of such a fiber material should be determined in consideration of the intended adsorption performance. For example, in order to remove granulocytes from blood, the cross-sectional diameter is preferably 4 μm or more, more preferably 5 μm to 10 μm. On the other hand, if a fiber having a cross-sectional diameter of 0.5 to 4 μm is used, it can be suitably used for removing lymphocytes. In addition, if fibers having a cross-sectional diameter of less than 0.5 μm are used, it is possible to increase the removal efficiency of the physiologically active substance. In addition, it is also possible to use a mixture of two or more types of fibers having different thicknesses. The diameters shown here are not only applied to columnar fibers, but are also applied to, for example, ellipses, rectangles and polygons. In those cases, the diameter is calculated assuming a perfect circle of the same area.
The thickness of the adsorption carrier is not limited, but is preferably from 0.1 mm to 10 cm for handling. For example, in the case of incorporating in a radial flow type module such as Tremixin manufactured by Toray, the thickness is preferably 1 cm or less because it is wound around a central pipe. These are determined by the handling method.

  The adsorbing material of the present invention is suitably used for medical material applications. Examples of the medical material application include, but are not limited to, a filler for a column for extracorporeal circulation of blood.

(Preparation of N, N-dimethyloctylaminated felt (DMOA felt))
Thirty-six sea-island composite fibers, each of which is made of a core-sheath composite, were obtained using the following components under the spinning conditions of a spinning speed of 800 m / min and a draw ratio of 3 times.
Island core component; Polypropylene island sheath component: Polystyrene 90wt%, Polypropylene 10wt%
Sea component: ethylene terephthalate unit as the main repeating unit, copolymerized polyester composite ratio (weight ratio) containing 3 wt% of 5-sodium sulfoisophthalic acid as a copolymer component; core: sheath: sea = 42: 43: 15
A nonwoven fabric (weight per unit area: 150 g / m ² ) made of 85 wt% of the fibers and 15 wt% of polypropylene fibers having a diameter of 20 μm was produced. Next, this non-woven fabric was treated with an aqueous solution of sodium hydroxide at 90 ° C. and 3 wt% to make ethylene terephthalate units as sea components as main repeating units, and a copolymer polyester containing 3 wt% of 5-sodium sulfoisophthalic acid as a copolymer component. By dissolving, a felt having a core-sheath fiber diameter of 5 μm was produced.
Next, 46 wt% of nitrobenzene, 46 wt% of sulfuric acid, 1 wt% of paraformaldehyde, 7 wt% of N-methylol-α-chloroacetamide (hereinafter abbreviated as NMCA) are mixed, stirred and dissolved at 10 ° C or less to prepare an NMCA reaction solution. did. The NMCA reaction solution was brought to 15 ° C., the NMCA reaction solution was added at a solid / liquid ratio of about 40 ml to 1 g of felt, and the reaction solution was reacted for 2 hours while maintaining the reaction solution at 15 to 20 ° C. in a water bath. During the 2 hours, the felt and the reaction solution were stirred by hand at 10 minute intervals. The felt was pulled up from the reaction solution, immersed in the same amount of nitrobenzene as the NMCA reaction solution, and shaken at 40 times / minute for 30 minutes with a seesaw type shaker (BIO CRAFFT, BC-700 type). Subsequently, the felt was lifted, immersed in methanol, and shaken for 30 minutes at 40 times / minute with a seesaw type shaker (BIO CRAFFT, BC-700). Repeated methanol washing after NMCA was repeated a total of 7 times. , NMCA felt was obtained.

Subsequently, 71 wt% dimethyl sulfoxide (DMSO), 22 wt% methanol, 6 wt% N, N-dimethyloctylamine (DMOA), and 1 wt% potassium iodide were mixed and dissolved to prepare a DMOA reaction solution. The DMOA reaction solution in the same amount as the NMCA reaction solution was heated to 50 ° C., the NMCA felt was immersed, and reacted in a 50 ° C. warm bath for 4 hours. After the DMOA, the felt is lifted from the reaction solution, immersed in the same amount of methanol as the NMCA reaction solution, and shaken at 40 times / minute for 30 minutes with a seesaw type shaker (BIO CRAFFT, BC-700 type). The methanol washing was repeated 7 times in total. Subsequently, the felt is pulled up, immersed in the same amount of water as the NMCA reaction solution, and washed with water by shaking for 30 minutes at 40 times / minute with a seesaw type shaker (BIO CRAFFT, BC-700 type). Repeated a total of 3 times to obtain a DMOA felt (hereinafter referred to as adsorption carrier 1).
(Saline treatment)
The adsorption carrier 1 was punched into a circle having a diameter of 1 cm, and packed in a column having an inner diameter of 1 cm and a length of 2 cm so that 10 sheets were stacked. A 20 ml 2M saline solution was circulated through this column at a flow rate of 30 ml / min for 30 minutes, and then the saline treatment was repeated for a new exchange of the 2M saline solution (hereinafter, this number of repetitions will be referred to as the number of saline treatment times). The adsorption carrier was taken out and rinsed with water to remove the saline solution attached to the adsorption carrier. As a sample used for measurement of the amount of residual iodine, a sample which had been completely dried by a vacuum dryer was used. As a sample used for the cytokine adsorption ability test, a sample immersed and stored in physiological saline was used.
(Measurement of residual iodine)
The amount of iodine remaining was measured at the Toray Research Center. A dried adsorption carrier was used as a measurement sample. In the method, first, the iodine residual amount of the adsorption carrier 1 was measured by a flask combustion method and potentiometric titration. Subsequently, the fluorescent X-ray intensity was measured by fluorescent X-ray analysis for this sample and the sample after the saline treatment, and the iodine residual amount of other samples was determined from the intensity ratio.
(Measurement of cytokine (IL-6) adsorption capacity)
Frozen fetal bovine serum (FBS) was immersed in a 56 ° C. warm bath, and after confirming that all were dissolved, it was further immersed in a 56 ° C. warm bath for 2 hours. IL-6 was added to this FBS so as to be 500 pg / ml. In Comparative Example 1, the adsorption carrier 1 was punched into a circle having a diameter of 1 cm, and the punched adsorption carrier 1 was immersed in FBS to which 0.95 ml of IL-6 was added. In Examples 2, 4 and Comparative Example 2, the above-mentioned saline treatment was performed a predetermined number of times, and then one adsorption carrier punched out into a circle having a diameter of 1 cm subjected to the saline solution treatment was treated with 0.95 ml of IL-6. It was immersed in the added FBS. Moreover, FBS which added IL-6 which does not add the adsorption carrier as a blank was prepared, and these were rotated with the rotator for 1 hour in a 37 degreeC incubator, respectively. The cytokine concentration in the supernatant was measured by ELISA. As the ELISA kit, a human IL-6 ELISA kit manufactured by Kamakura Technoscience was used. As the notation of the adsorption capacity, the difference between the IL-6 concentration in the blank FBS and the IL-6 concentration in the FBS after the adsorption test with the adsorption carrier was expressed as a percentage (IL-6 adsorption rate) with respect to the blank concentration.
[Example 1]
It was 1.4 wt% as a result of measuring iodine residual amount after performing the salt solution process with respect to the adsorption carrier 1 twice.
[Example 2]
After subjecting the adsorption carrier 1 to the saline treatment three times, the IL-6 adsorption rate was measured. As a result, the IL-6 adsorption capacity was 82%.
[Example 3]
It was 0.027 wt% as a result of measuring the iodine residual amount after performing the salt solution process 6 times with respect to the adsorption carrier 1. FIG.
[Example 4]
After the saline treatment was performed 12 times on the adsorption carrier 1, the amount of iodine remaining and the IL-6 adsorption rate were measured. As a result, the amount of iodine remaining in the felt was below the detection limit (0.08 wt% or less). Met. The IL-6 adsorption capacity was about 84%.
[Comparative Example 1] The iodine residual amount and the IL-6 adsorption rate were measured without subjecting the adsorption carrier 1 to the saline treatment. As a result, the iodine residual amount remaining in the felt was 9.6 wt%. . The IL-6 adsorption capacity was about 66%.
[Comparative Example 2] After washing with saline once, the IL-6 adsorption rate of this adsorbent carrier 1 was measured. As a result, the IL-6 adsorption capacity was 76%.
[Comparative Example 3]
The adsorption carrier 1 was punched into a circle having a diameter of 1 cm, and packed in a column having an inner diameter of 1 cm and a length of 2 cm so that 10 sheets were stacked. This column was circulated with 20 ml of 2M potassium iodide aqueous solution at a flow rate of 30 ml / min for 30 minutes, and then the potassium iodide aqueous solution treatment of newly exchanging the 2M potassium iodide aqueous solution was repeated 12 times. Replaced with ions. The adsorption carrier was taken out and rinsed with water to remove potassium iodide attached to the adsorption carrier. Samples to be used in the cytokine adsorption capacity test were immersed in physiological saline and stored.

  The result of measuring the IL-6 adsorption rate of this adsorption carrier was 20%.

  As shown in FIG. 1, in this adsorption carrier 1, when the number of times of the saline treatment is 2 times or more (residual iodine amount: 1.4 wt%), the adsorption carrier produced under these conditions stably adsorbs 80% or more of IL-6. It turns out to get a rate.

The adsorption carrier of the present invention can be suitably used for various blood processing columns. It is particularly suitable for removing both leukocytes and cancer cells unnecessary for the human body that are present in excess and physiologically active substances such as cytokines, and is useful for blood treatment and treatment of autoimmune diseases, cancer, allergies, etc. It is. This material is also suitable for affinity chromatography columns, therapeutic blood columns, especially extracorporeal circulation columns in the form of petri dishes, bottles, membranes, fibers, hollow fibers, granules, or molded articles using these. Can be used for

Relationship between the number of saline treatments, the amount of iodine remaining, and the IL-6 adsorption rate

Claims (8)

  An adsorption carrier containing iodine in an amount exceeding 1.4 wt%, wherein the iodine content after the iodine removal treatment is 1.4 wt% or less.   The adsorption carrier according to claim 1, which has a basic functional group.   The adsorption carrier according to claim 2, wherein the basic functional group is a quaternary ammonium group.   The adsorption carrier according to any one of claims 1 to 3, which has a felt form.   The adsorption carrier according to any one of claims 1 to 4, which is used for medical materials.   A method for producing an adsorption carrier, characterized in that an adsorption carrier having an iodine content exceeding 1.4 wt% is treated with a liquid containing a halogen compound excluding an iodine compound so that the iodine content is 1.4 wt% or less.   The method for producing an adsorption carrier according to claim 6, wherein the adsorption carrier into which a functional group is introduced using iodine as a reaction reagent is treated with a liquid containing a halogen compound excluding an iodine compound.   The method for producing an adsorption carrier according to claim 6 or 7, wherein the halogen compound excluding the iodine compound is a chlorine compound.

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