JP2007023109A - Maleimidyl group-bearing polymer particles and method for producing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide polymer particles having a sufficient amount of maleimidyl groups and a method for producing the same. <P>SOLUTION: In the maleimidyl group including polymer particles, a maleimidyl group represented by chemical formula (1) is introduced in the cross-linked polymer prepared by co-polymerization of a monomer mixture of (meth)acrylate, styrene monomer and a crosslinking agent. The cross-linked polymer particles are produced via the step where the monomer mixture is allowed to copolymerize in the presence of non-polymerizing solvent in an amount of 35mass% based on the total of the monomer mixture and the non-polymerizing solvent and the step where the non-polymerizing solvent is removed from the solvent-including particles. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  TECHNICAL FIELD The present invention relates to maleimidyl group-containing polymer particles and a method for producing the same, and particularly for uses such as diagnostic agents and pharmaceutical carriers, chromatography carriers, viscosity modifiers, resin molding materials, paint additives, crosslinking / curing agents, and cosmetic additives. The present invention relates to a maleimidyl group-containing polymer particle that can be suitably used and a method for producing the same.

  Functional polymer particles such as ion exchange resins and chelate resins have been widely used as carriers for various chemical substances. Such functional polymer particles have various reactive groups on the surface of the insoluble polymer. Conventionally, functional polymers into which various active hydrogen-containing groups such as carboxyl groups, hydroxyl groups, and primary and secondary amino groups have been introduced have been used for various applications.

    In recent years, biomolecules such as nucleic acids, peptides, and antibodies, or biomolecule-like synthetic molecules are immobilized on polymer particles while retaining activity, and are used for affinity chromatography, diagnostic drugs, test drugs, and the like. When used as a carrier particle having higher selectivity in this way, a maleimidyl group (maleimide) that can be reliably supported by forming a selective and stable bond with an SH group in a biomolecule or a biomolecule-like synthetic molecule. The group having the group) was desired. In such an application, the polymer carrier needs to carry a desired amount of a substance uniformly and reliably, and has a good dispersibility in an aqueous solvent.

There are few known examples of polymer particles having maleimidyl groups. For example, a fine particle in which a phospholipid film containing a maleimidyl group is present on the surface is disclosed (for example, see Patent Document 1).
However, in this method, a maleimidyl group-containing phospholipid must be prepared in advance by reacting a maleimidyl group-containing compound such as N- (6-maleimidocaproyloxy) succinimide with a phospholipid and purifying it by a method such as column chromatography. Don't be. Furthermore, a phospholipid film containing maleimidyl groups is produced through multi-step processing, and this is formed on a core particle such as a magnetic particle, so the process is extremely complicated, and the phospholipid containing core particles and maleimidyl groups There is a disadvantage that it is physically weak because it is not chemically bonded to the film.
In addition, there is an example in which polystyrene particles containing a maleimidyl group are published (for example, see Non-Patent Document 1). However, the maleimidyl group is a functional group having strong hydrophobicity, and its dispersibility in an aqueous medium is very poor. Furthermore, the amount of maleimidyl groups introduced into the polymer particles was insufficient.
JP-A-11-106391 2001/2002 product catalog, Fulka, p. 909

  This invention is made | formed in view of the said problem, and it aims at providing the polymer particle which has sufficient maleimidyl group, and its manufacturing method.

That is, the present invention
<1> Maleidyl group-containing polymer particles in which a maleimidyl group represented by the following chemical formula (1) is introduced into a crosslinked polymer particle obtained by copolymerizing a monomer mixture containing a (meth) acrylate, a styrene monomer and a crosslinking agent. The crosslinked polymer particles comprise the monomer mixture, the monomer mixture is soluble, and the copolymer of the (meth) acrylate and the styrenic monomer is hardly soluble, and the monomer mixture. And a step of obtaining non-polymerizable solvent-containing particles by copolymerization in a state where the content of the non-polymerizable solvent is 35% by mass or more with respect to the total amount, and non-polymerizable from the non-polymerizable solvent-containing particles Maleimidyl group-containing polymer particles obtained through a step of removing the solvent.

(In the formula, m and n represent an integer of 1 or more.)

  <2> The maleidyl group-containing polymer particle according to <1>, wherein the non-polymerizable solvent is at least one selected from hydrocarbons having a boiling point of 100 ° C. or higher, long-chain alcohols, long-chain carboxylic acids, and long-chain esters. is there.

  <3> The maleimidyl group-containing polymer particle according to <1> or <2>, wherein an average pore radius by a mercury intrusion method is 0.005 to 0.1 μm.

  <4> Non-polymerization of a monomer mixture containing a (meth) acrylate, a styrene monomer, and a crosslinking agent, wherein the monomer mixture exhibits solubility and the copolymer of the (meth) acrylate and the styrene monomer exhibits poor solubility. A step of obtaining non-polymerizable solvent-containing particles by copolymerization in a state where the content of the non-polymerizable solvent is 35% by mass or more based on the total amount of the polymerizable solvent and the monomer mixture; A method for producing maleimidyl group-containing polymer particles, comprising: obtaining a crosslinked polymer particle by removing a non-polymerizable solvent from the contained particle; and introducing a maleimidyl group represented by the following chemical formula (1) into the crosslinked polymer particle: It is.

(In the formula, m and n represent an integer of 1 or more.)

  <5> The maleidyl group-containing polymer particle according to <4>, wherein the non-polymerizable solvent is at least one selected from a hydrocarbon having a boiling point of 100 ° C. or higher, a long-chain alcohol, a long-chain carboxylic acid, and a long-chain ester. It is a manufacturing method.

  ADVANTAGE OF THE INVENTION According to this invention, the polymer particle which has sufficient maleimidyl group, and its manufacturing method can be provided.

  The maleimidyl group-containing polymer particle of the present invention is obtained by introducing a maleimidyl group represented by the following chemical formula (1) into a crosslinked polymer particle obtained by copolymerizing a monomer mixture containing (meth) acrylate, a styrene monomer and a crosslinking agent. A non-polymerizable solvent in which the crosslinked polymer particles are soluble in the monomer mixture, the monomer mixture is soluble, and the copolymer of the (meth) acrylate and the styrenic monomer is insoluble. From the step of obtaining non-polymerizable solvent-containing particles by copolymerizing in a state where the content of the non-polymerizable solvent is 35% by mass or more with respect to the total amount with the monomer mixture, And a step of removing the non-polymerizable solvent.

  In the formula, m and n represent an integer of 1 or more.

  Also, the method for producing maleimidyl group-containing polymer particles of the present invention includes a monomer mixture containing (meth) acrylate, a styrenic monomer and a crosslinking agent, wherein the monomer mixture exhibits solubility and the (meth) acrylate and the styrenic monomer. The non-polymerizable solvent is copolymerized in a state where the content of the non-polymerizable solvent is 35% by mass or more based on the total amount of the non-polymerizable solvent and the monomer mixture. A step of obtaining contained particles, a step of removing non-polymerizable solvent from the non-polymerizable solvent-containing particles to obtain crosslinked polymer particles, and a step of introducing a maleimidyl group represented by the following chemical formula (1) into the crosslinked polymer particles And.

  In the formula, m and n represent an integer of 1 or more.

By the way, in preparing polymer particles, it is known that a monomer mixture solution dissolves (indicates solubility), but a polymer that is a polymer of the monomer mixture solution is polymerized by adding a hardly soluble solvent (for example, J Polymer Sci., Part A, vol. 2, p.835-843 (1964)), these methods are techniques aimed at making the polymer particles porous. However, the crosslinked polymer particles according to the present invention have an average pore radius of 0.1 μm or less when the pore diameter is measured by a mercury intrusion method, and at least the surface of the particle is clear by observation with a scanning electron microscope (SEM). Since the structure is not formed, it does not seem that the specific surface area increased and the amount of maleimidyl groups increased.
Although the effect of the present invention is not well understood, it is considered that the chemical or physical properties of the particle surface have changed, and the reaction between the OH group at the particle end and hydroxymaleimide has been carried out efficiently.

  Here, “(meth) acrylate” means “acrylic ester” and “methacrylic ester” as commonly used. (Poly) ethylene glycol means ethylene glycol and polyethylene glycol.

  The present invention will be specifically described below.

<Maleimidyl group-containing polymer particles>
The maleimidyl group-containing polymer particles of the present invention are those in which the maleimidyl group represented by the chemical formula (1) is introduced into the crosslinked polymer particles.

  In the general formula (1), the maleimidyl group is chemically bonded to the crosslinked polymer particle via an ethylene oxide chain, and m of the ethylene oxide chain represents an integer of 1 or more. The ethylene oxide chain can be obtained by reacting (poly) ethylene glycol with cross-linked polymer particles and chemically bonding them, or (meth) acrylate having (poly) ethylene glycol introduced therein or a precursor thereof. (Meth) acrylate introduced with may be copolymerized and directly introduced.

The maleimidyl group-containing polymer particle of the present invention may have a maleimidyl group represented by the chemical formula (1) on the surface of the crosslinked polymer particle or on the inside of the crosslinked polymer particle. And both of them may be included.
Generally, m is preferably 2 or more from the viewpoint of obtaining good dispersibility in an aqueous solvent.
Further, since hydroxyalkylmaleimide represented by hydroxymethylmaleimide is suitable as a raw material compound for introducing a maleimidyl group into polymer particles into which (poly) ethylene glycol is introduced, n is preferably 1 or more.

  The maleimidyl group-containing polymer particles of the present invention preferably contain 0.001 mmol / g or more of the maleimidyl group represented by the chemical formula (1). From the viewpoint of increasing the reaction amount per unit mass as much as possible, 0.01 mmol / g or more is preferable, and 0.02 mmol / g or more is particularly preferable. By setting it to 0.02 mmol / g or more, the reaction rate with the SH group-containing compound can be increased.

  The maleimidyl group-containing polymer particle of the present invention has a property of good dispersibility in an aqueous solvent since a hydrophilic ethylene glycol or polyethylene glycol chain exists between the maleimidyl group and the crosslinked polymer particle. ing.

  Thus, the maleimidyl group-containing polymer particles of the present invention have good dispersibility in an aqueous solvent, and the support of the maleimidyl group-containing polymer particles is between the SH groups in the biomolecules or biomolecule-like synthetic molecules. It forms a selective and stable bond and can be used as a carrier particle with high selectivity.

The maleimidyl group-containing polymer particles of the present invention are not particularly limited, and can be selected according to various applications. Generally, the average particle diameter thereof is 1 to 1000 μm. From the viewpoint of ease, it is preferably 3 to 500 μm, and particularly preferably 5 to 100 μm.
By setting the average particle size in the range of 1 to 1000 μm, solid-liquid separation characteristics by filtration or centrifugation can be improved while maintaining a sufficient reaction amount per unit mass.

  The average pore radius by the mercury intrusion method of the maleimidyl group-containing polymer particles of the present invention is preferably 0.005 to 0.1 μm, and more preferably 0.01 to 0.05 μm or less. When the average pore radius is 0.005 to 0.1 μm, maleimidyl group-containing polymer particles having a high introduction amount of maleimidyl groups and high peptide reactivity can be obtained.

<Crosslinked polymer particles>
The crosslinked polymer particle according to the present invention comprises a monomer mixture containing (meth) acrylate, a styrene monomer and a crosslinking agent, the monomer mixture is soluble, and a copolymer of (meth) acrylate and a styrene monomer is hardly soluble. A step of obtaining non-polymerizable solvent-containing particles by copolymerization in a state where the content of the non-polymerizable solvent is 35% by mass or more with respect to the total amount of the non-polymerizable solvent and the monomer mixture, And a step of removing the non-polymerizable solvent from the solvent-containing particles.

  In the (meth) acrylate, the alkyl group of the alcohol residue of the (meth) acrylic acid ester is an alkyl group having 1 to 18 carbon atoms, which may be unsubstituted or substituted, for example, methyl, Ethyl, n-propyl, isopropyl, n-butyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, n-octyl, nonyl, decyl, undecyl, dodecyl, benzyl, hydroxyethyl, hydroxy group such as dihydropyran And hydroxyethyl, polyoxyethylene and the like protected with a protective group. When the polymer is hydrolyzed to introduce (poly) ethylene glycol, t-butyl (meth) acrylate is preferably present as one component.

  The styrenic monomer is a vinyl group-containing monomer having an aryl group having 6 to 12 carbon atoms, which may be unsubstituted or substituted. For example, phenyl, naphthyl, tolyl, pn-octyl Although oxyphenyl etc. are mentioned, phenyl is preferable.

Examples of the substituent include an alkyl group, an alkoxy group, a halogen atom, and an aryl group.
As this alkyl group, it is synonymous with the case of the above-mentioned alkyl group. Examples of the alkoxy group include methoxy, ethoxy, propoxy, butoxy and the like, and methoxy and ethoxy are preferable.
Examples of the halogen atom include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom, and a fluorine atom and a chlorine atom are preferable.
As this aryl group, it is synonymous with the case of the above-mentioned aryl group.

  Examples of the crosslinking agent include divinylbenzene, ethylene glycol di (meth) acrylate, diethylene glycol di (meth) acrylate, glycidyl (meth) acrylate, and 2-([1′-methylpropylideneamino] carboxyamino) ethyl (meth) acrylate. Etc.

  The ratio of the content of (meth) acrylate and styrene monomer in the monomer mixture is preferably (meth) acrylate: styrene monomer = 90: 10 to 10:90, and 25:75 to 75:25 in molar ratio. Further preferred. Further, the content of the crosslinking agent in the monomer mixture is preferably 0.5 to 30 parts, more preferably 1.0 to 20 parts, with respect to 100 parts of the (meth) acrylate, the styrene monomer, and the total amount.

  In addition, the crosslinked polymer particles can control the amount of polyethylene glycol introduced into the particles by changing the copolymerization ratio of the polymer component of the monomer, and therefore, the maleimidyl groups in the maleimidyl group-containing polymer particles can be controlled. Can be controlled.

The non-polymerizable solvent used in the present invention is one in which the monomer mixture is soluble and the copolymer of (meth) acrylate and styrene monomer is insoluble.
In the present invention, “the monomer mixture is soluble” means that the amount of the monomer mixture dissolved in the non-polymerizable solvent at 15 ° C. is 2.5 g / g or more, and “(meth) acrylate and styrene”. “A copolymer with a monomer exhibits poor solubility” means that the amount of the copolymer dissolved in a non-polymerizable solvent is 2.5 g / g or less.
The non-polymerizable solvent is not particularly limited, but is preferably at least one selected from hydrocarbons having a boiling point of 100 ° C. or higher, long-chain alcohols, long-chain carboxylic acids and long-chain esters. The non-polymerizable solvent may be used alone or in combination of two or more.
Non-polymerizable solvents include hydrocarbons such as decane, dodecane and hexadecane, long-chain alcohols such as amyl alcohol, octanol, decanol, undecanol, lauryl alcohol and stearyl alcohol, long-chain carboxylic acids such as lauric acid and stearic acid, laurin Long-chain esters such as ethyl acrylate, butyl laurate, ethyl stearate, and butyl stearate. In view of increasing the polymerization temperature, the boiling point is preferably 100 ° C or higher. Furthermore, the melting point is preferably 10 ° C. or higher in order to suppress the aggregation of the particles after polymerization and improve the workability of the post-treatment.
Specific examples include hexadecane, lauryl alcohol, stearyl alcohol, and butyl stearate.

The copolymerization reaction of the monomer mixture is performed in a state where the content of the nonpolymerizable solvent is 35% by mass or more with respect to the total amount of the monomer mixture and the nonpolymerizable solvent. Preferably it is 38 mass% or more, More preferably, it is 42 mass% or more. When the content of the non-polymerizable solvent is less than 35% by mass, the amount of maleimidyl groups introduced into the particles may be insufficient. Further, when (poly) ethylene glycol is to be introduced into the crosslinked polymer particles by transesterification as described later, the amount of hydroxyl group introduced becomes insufficient, and a larger amount of catalyst and reaction time may be required.
Crosslinked polymer particles (non-polymerizable solvent-containing particles) containing a non-polymerizable solvent are obtained by copolymerization reaction of the monomer mixture.

  In the present invention, known methods can be used to obtain crosslinked polymer particles. For example, a suspension polymerization method, an emulsion polymerization method, a dispersion polymerization method, a seed polymerization method and the like are preferably used. Furthermore, suspension polymerization can be performed using an emulsification method known as a membrane emulsification method.

  The non-polymerizable solvent is removed from the non-polymerizable solvent-containing particles obtained as described above. The non-polymerizable solvent is extracted and removed using a solvent such as ethanol, isopropanol, acetone, or toluene. In this case, a Soxhlet extractor or the like can be used.

For the copolymerization reaction of the monomer mixture, a polymerization initiator well known to those skilled in the art can be used as necessary.
Specifically, organic peroxides such as diacyl peroxide, ketone peroxide and alkyl hydroperoxide; inorganic peroxides such as hydrogen peroxide and ozone; and azobisisobutyronitrile (AIBN; Wako Pure) (Available from Yakuhin as V-60), 2,2′-azobis (2-methylbutyronitrile) (available as V-59 from Wako Pure Chemical Industries, Ltd.) and 2,2′-azobis (2,4- Oil-soluble azo organic compounds such as dimethylvaleronitrile (available as V-65 from Wako Pure Chemical Industries); 2,2′-azobis (
2-Amidinopropane) dioic acid salt (available as V-50 from Wako Pure Chemical Industries), 2,2'-
Azobis [2-methyl-N- (2-hydroxyethyl) propionamide] (available as VA-086 from Wako Pure Chemical Industries) and 2,2′-azobis [2- (2-imidazolin-2-yl) propane ] Water-soluble azo organic compounds such as diacid salts (available as VA-044 from Wako Pure Chemical Industries). If polymerization initiators are used, they are used in an amount sufficient to initiate polymerization well. Such amounts are well known to those skilled in the art. In general, it is preferably used in an amount of 0.1 to 5.0% by mass.

  Furthermore, for the purpose of coloring the crosslinked polymer particles, known dyes, pigments, carbon black, magnetic powder and the like can be added.

  Since the crosslinked polymer particles in the present invention do not have a special reactive functional group, there is an advantage that there is no or very little adverse effect on the physical properties of the particles due to reaction with additives and solvents and remaining.

<Hydroxyl-containing polymer particles>
The hydroxyl group-containing polymer particles used in the present invention are polymer particles obtained by introducing (poly) ethylene glycol into crosslinked polymer particles.
Further, (poly) ethylene glycol means a diol compound represented by a general formula of HO (CH ₂ CH ₂ O) _n H. n is synonymous with n number of the said Chemical formula (1).
The method for introducing (poly) ethylene glycol into the crosslinked polymer particles may be any method and can be appropriately selected depending on the purpose.
For example, the crosslinked polymer particles are hydrolyzed with acid or alkali, the ester part of the (meth) acrylate residue is once converted into a carboxyl group, and a diol compound such as ethylene glycol or polyethylene glycol in the presence of a catalyst for esterification reaction Hydroxyl group-containing polymer particles may be obtained by ester reaction with hydroxyl groups of the polymer, or crosslinked polymer particles may be obtained by directly transesterifying a crosslinked polymer particle with a diol compound such as ethylene glycol or polyethylene glycol using an appropriate transesterification catalyst. It is good also as a hydroxyl-containing polymer particle combining with this.

In the present invention, the hydroxyl group-containing polymer particles are preferably obtained by reacting a carboxyl group obtained by hydrolyzing the crosslinked polymer particles with (poly) ethylene glycol. Furthermore, it is also preferred that the crosslinked polymer particles be a hydroxyl group-containing polymer obtained by direct transesterification with (poly) ethylene glycol in the presence of a metal-containing transesterification catalyst.
In order to facilitate the transesterification, it is preferable to react an equal amount or more of (poly) ethylene glycol with respect to the total molar amount of the functional groups contained in the crosslinked polymer particles. Since the group does not transesterify, the amount may be equal or less.
When the crosslinking degree of the crosslinked polymer particles is high, a non-reactive solvent capable of swelling the polymer particles can be added as a cosolvent for the purpose of promoting the reaction. For example, aromatic compounds such as toluene, xylene, mesitylene, diethylbenzene, cyclohexylbenzene, dimethylnaphthalene, cymene, dimethoxybenzene, ether compounds such as diethylene glycol dimethyl ether, diethylene glycol dibutyl ether, diphenyl ether, dibenzyl ether, phenetol, butyl phenyl ether, acetophenone , Ketone compounds such as isophorone, diisobutyl ketone, cyclohexanone, and methylcyclohexanone, and halogen compounds such as dichlorobenzene, chlorotoluene, and dichlorotoluene. Any nonreactive solvent that achieves the above-described purpose can be used. Among these, aromatic compounds such as toluene, xylene, mesitylene, and dimethoxybenzene are particularly preferable in terms of reactivity.
The hydroxyl group-containing polymer particles thus obtained are preferably used as a raw material for the maleimidyl group-containing polymer particles of the present invention.

In the present invention, the reaction ratio between the crosslinked polymer particles and (poly) ethylene glycol varies depending on the combination of carboxyl groups or ester portions of the crosslinked polymer particles and the amount of these functional groups introduced, but basically the crosslinked polymer particles. About 2 to 10 times the amount of the (poly) ethylene glycol by weight, in the non-reactive solvent used as necessary, in the esterification reaction, 80 ° C. to 150 ° C., in the transesterification reaction 120 ° C. Then, hydroxyl group-containing polymer particles can be obtained by heating for about 5 to 24 hours under the reaction conditions of 200 ° C. for reaction.
Here, from the viewpoint of reproducibility of the amount of introduced hydroxyl group, it is desirable to classify the particles before the reaction in advance and to react after the particle diameters are made uniform. The hydroxyl group-containing polymer particles thus obtained are diluted and dispersed in a solvent such as methanol, filtered, further washed with water and / or solvent, and then powdered by ordinary means such as spray drying, reduced pressure drying, freeze drying and the like. It can be isolated and subjected to the maleimidyl group introduction reaction of the next step, or it is used in the reaction system of the next step in a wet state without passing through the drying step, and the solvent is removed from the reaction system before or simultaneously with the maleimidyl group introduction reaction. You can also.

As the esterification reaction catalyst to be added in the present invention, a known esterification reaction catalyst can be preferably used. For example, inorganic acids such as sulfuric acid, hydrochloric acid, nitric acid and phosphoric acid, and organic acids such as p-toluenesulfonic acid, trichloroacetic acid and acetic acid can be used. A condensing agent such as DCC (N, N-dicyclohexylcarbodiimide) can also be suitably used.
As the metal-containing transesterification catalyst added in the present invention, a so-called transesterification catalyst used at the time of polyester synthesis can be suitably used.
Examples of the metal-containing transesterification catalyst include lead acetate, zinc acetate, zinc acetylacetonate, cadmium acetate, manganese acetate, manganese acetylacetonate, cobalt acetate, cobalt acetylacetonate, nickel acetate, nickel acetylacetonate, Zirconium acetate, zirconium acetylacetonate, zirconium tetra-n-butoxide, titanium acetate, tetrabutoxy titanate, tetraisopropoxy titanate, titanium oxyacetylacetonate, transition metal compounds such as iron acetate, iron acetylacetonate, niobium acetate, dibutyltin Oxides, monobutylhydroxytin oxide, dibutyltin dilaurate, antimony trioxide, germanium oxide, bismuth oxide, bismuth acetate, etc. Type metal compound, and the like.
Also, alkali metal compounds or alkaline earth metal compounds such as magnesium acetate, lithium acetate, calcium acetate, potassium acetate, potassium carbonate, cesium carbonate, and rare earths such as lanthanum acetate, samarium acetate, europium acetate, erbium acetate, ytterbium acetate Compounds can also be used.

  These catalysts may be used alone or in combination of two or more. As the metal-containing transesterification catalyst, titanium tetraalkoxide is more preferable in terms of solubility in a solvent and reactivity. Examples of the titanium tetraalkoxide include tetra-n-propoxy titanium, tetraisopropoxy titanium, tetra-n-butoxy titanium, tetraisobutoxy titanium, tetra-t-butoxy titanium, tetra-secondary butoxy titanium, and tetra-n. -Amyloxytitanium, triisoamyloxyisopropoxytitanium and the like.

The added amount (content) of the catalyst to the crosslinked polymer particles is usually 0.01 to 50 parts by mass with respect to 100 parts by mass of the total particle weight. Preferably it is 0.1-20 mass parts, More preferably, it is 0.5-10 mass parts.
By making a catalyst into the range of 0.01-50 mass parts, it becomes easy to introduce | transduce a hydroxyl group, and the removal of the catalyst after reaction can also be advanced advantageously.

<Manufacturing method of maleimidyl group-containing polymer particles>
The maleimidyl group-containing polymer particles of the present invention can be obtained by reacting hydroxymethylmaleimide with hydroxyl group-containing polymer particles obtained by introducing (poly) ethylene glycol. In the method for producing maleimidyl group-containing polymer particles of the present invention, the step of introducing the maleimidyl group represented by the chemical formula (1) into the crosslinked polymer particles includes the step of obtaining the hydroxyl group-containing polymer particles from the crosslinked polymer particles described above, Obtaining maleimidyl group-containing polymer particles from the polymer particles.

The reaction between the hydroxyl group-containing polymer particles and hydroxymethylmaleimide is a form in which one molecule of water is removed and condensed, and can be carried out by a conventionally known reaction, but in particular, it is preferably carried out in the presence of an etherification catalyst.
For this reaction, an acidic or basic known etherification catalyst can be preferably used. For example, as the basic catalyst, hydroxides, oxides, carbonates, bicarbonates, and the like of alkali metals and alkaline earth metals can be used, and they can be used alone or in combination of two or more. As the acidic catalyst, inorganic acids such as sulfuric acid, hydrochloric acid, nitric acid and phosphoric acid and organic acids such as p-toluenesulfonic acid, trichloroacetic acid and acetic acid can be used, and these may be in the form of hydrates. A hydrotalcite-type solid catalyst can also be used.
The amount of the basic catalyst or acidic catalyst used is 0.01 to 40% by mass, preferably 0.1 to 15% by mass, based on hydroxymethylmaleimide, as the corresponding base or acid. In the case of a solid catalyst, it is 0.001-100 mass% with respect to hydroxymethylmaleimide, Preferably it is 0.1-50 mass%. The catalyst or the like may be used in a state of being uniformly dissolved in the reaction solution or in an insoluble state, but the amount used can be reduced in the state of uniform dissolution. On the other hand, in the insoluble state, the catalyst can be easily separated and recovered from the reaction solution by a conventional method after the reaction.

  The reaction rate of the hydroxyl group-containing polymer particles and hydroxymethylmaleimide varies depending on the combination of the ester portion of the hydroxyl group-containing polymer particles and the amount of these functional groups introduced, but is basically about 0 with respect to the weight of the hydroxyl group-containing polymer particles. Maleimide-containing polymer particles can be obtained by using 1 to 10 times the amount of the hydroxymethylmaleimide.

  Further, the reaction temperature and reaction time in this reaction vary depending on the combination of the ester portion of the hydroxyl group-containing polymer particles, the amount of these functional groups introduced, the ethylene oxide chain length, etc., and can be appropriately selected. Generally, maleimide-containing polymer particles can be obtained by heating and reacting at 80 to 180 ° C. for 2 to 20 hours.

  As the solvent for the etherification reaction, any solvent can be used as long as it does not inhibit the reaction. For example, toluene, xylene, and mesitylene are mentioned.

  In view of the reproducibility of the amount of maleimidyl group introduced, it is desirable to classify the particles in advance before the reaction and to react after aligning the particle diameters. The maleimide-containing polymer particles thus obtained are diluted and dispersed in a solvent such as methanol, filtered, and further washed with water and / or solvent, and then powdered by ordinary means such as spray drying, reduced pressure drying, freeze drying and the like. It can be isolated.

  Further, the production method of the present invention can be reacted not only in the air but also under pressure, but these other reaction conditions are applied as necessary and are particularly limited. Yes.

  The amount of maleimidyl groups in the maleimidyl group-containing polymer particles obtained by the above operation is obtained by reacting the maleimidyl groups present in the particles with an excess amount of 2-mercaptoethylamine and measuring the amount of 2-mercaptoamine remaining after the reaction. Conversely, the amount of maleimidyl groups in the particles can be determined. Specific examples thereof are described, for example, on page 80 of the first part of the general catalog (published March 29, 2002) of Dojindo Laboratories.

  Useful maleimidyl group-containing polymer particles can be obtained by the production method of the present invention, such as diagnostic agents and pharmaceutical carriers, chromatography carriers, viscosity modifiers, resin molding materials, paint additives, crosslinking / curing agents, and cosmetic additives. It can be suitably used for various applications.

  EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, this invention is not limited to these Examples. In addition, in an Example, a part shows a mass part.

(Synthesis of crosslinked polymer particle 1)
Styrene (manufactured by Wako Pure Chemical Industries, Ltd.) 35 parts, t-butyl methacrylate (manufactured by Wako Pure Chemical Industries, Ltd.) 47 parts, divinylbenzene (purity 55%) 12 parts, lauryl alcohol (manufactured by Wako Pure Chemical Industries, Ltd.) Suspension polymerization was performed using 75 parts and 2.0 parts of a polymerization initiator V601 (manufactured by Wako Pure Chemical Industries, Ltd.). After washing / drying, Soxhlet extraction with ethanol was performed for 10 hours, and then classification was performed to obtain an average. Crosslinked polymer particles having a particle size of 55 μm were obtained. The obtained particles were washed with ion-exchanged water and a solvent, and then isolated and dried to obtain crosslinked polymer particles 1. The content of the non-polymerizable solvent relative to the total amount of the monomer mixture (styrene, t-butyl methacrylate and divinylbenzene) and the non-polymerizable solvent (lauryl alcohol) was 44%.

(Synthesis of crosslinked polymer particles 2)
Styrene (manufactured by Wako Pure Chemical Industries, Ltd.) 35 parts, t-butyl methacrylate (manufactured by Wako Pure Chemical Industries, Ltd.) 47 parts, divinylbenzene (purity 55%) 12 parts, hexadecane (manufactured by Wako Pure Chemical Industries, Ltd.) 75 Suspension polymerization using 2.0 parts by weight and polymerization initiator V601 (manufactured by Wako Pure Chemical Industries, Ltd.), washing / drying, and Soxhlet extraction with ethanol for 10 hours, followed by classification and average particle size Crosslinked polymer particles having a diameter of 55 μm were obtained. The obtained particles were washed with ion-exchanged water and a solvent, and then isolated and dried to obtain crosslinked polymer particles 2. The content of the non-polymerizable solvent relative to the total amount of the monomer mixture and the non-polymerizable solvent was 44%.

(Synthesis of crosslinked polymer particles 3)
Styrene (made by Wako Pure Chemical Industries, Ltd.) 35 parts, t-butyl methacrylate (made by Wako Pure Chemical Industries, Ltd.) 47 parts, divinylbenzene (purity 55%) 12 parts, stearic acid butyl ester (Wako Pure Chemical Industries, Ltd.) Suspension polymerization was performed using 75 parts of the product and 2.0 parts of polymerization initiator V601 (manufactured by Wako Pure Chemical Industries, Ltd.), washing / drying, and Soxhlet extraction with ethanol for 10 hours, followed by classification operation. As a result, crosslinked polymer particles having an average particle diameter of 55 μm were obtained. The obtained particles were washed with ion-exchanged water and a solvent, and then isolated and dried to obtain crosslinked polymer particles 3. The content of the non-polymerizable solvent relative to the total amount of the monomer mixture and the non-polymerizable solvent was 44%.

(Synthesis of crosslinked polymer particles 4)
Styrene (manufactured by Wako Pure Chemical Industries, Ltd.) 35 parts, t-butyl methacrylate (manufactured by Wako Pure Chemical Industries, Ltd.) 47 parts, divinylbenzene (purity 55%) 12 parts, stearic acid (manufactured by Wako Pure Chemical Industries, Ltd.) Suspension polymerization was performed using 75 parts and 2.0 parts of a polymerization initiator V601 (manufactured by Wako Pure Chemical Industries, Ltd.). Crosslinked polymer particles having a particle size of 55 μm were obtained. The obtained particles were washed with ion-exchanged water and a solvent, and then isolated and dried to obtain crosslinked polymer particles 4. The content of the non-polymerizable solvent relative to the total amount of the monomer mixture and the non-polymerizable solvent was 44%.

(Synthesis of Comparative Crosslinked Polymer Particles 5)
Except for not adding lauryl alcohol, the same operation as in the case of the crosslinked polymer particles 1 was performed to obtain crosslinked polymer particles having an average particle size of 55 μm. The obtained particles were washed with ion-exchanged water and a solvent, and then isolated and dried to obtain crosslinked polymer particles 5. The content of the non-polymerizable solvent relative to the total amount of the monomer mixture and the non-polymerizable solvent was 0%.

(Synthesis of Comparative Crosslinked Polymer Particles 6)
Styrene (manufactured by Wako Pure Chemical Industries, Ltd.) 35 parts, t-butyl methacrylate (manufactured by Wako Pure Chemical Industries, Ltd.) 47 parts, divinylbenzene (purity 55%) 12 parts, lauryl alcohol (manufactured by Wako Pure Chemical Industries, Ltd.) Suspension polymerization was performed using 15 parts and 2.0 parts of a polymerization initiator V601 (manufactured by Wako Pure Chemical Industries, Ltd.). After washing / drying, Soxhlet extraction with ethanol was performed for 10 hours, and then classification operation was performed. Crosslinked polymer particles having a particle size of 55 μm were obtained. The obtained particles were washed with ion-exchanged water and a solvent, and then isolated and dried to obtain crosslinked polymer particles 6. The content of the non-polymerizable solvent relative to the total amount of the monomer mixture (styrene, t-butyl methacrylate and divinylbenzene) and the non-polymerizable solvent (lauryl alcohol) was 14%.

(Synthesis of hydroxymethylmaleimide)
When 24 parts of maleimide (manufactured by Aldrich Co., Ltd.), 21 parts of 35% HCOH (manufactured by Wako Pure Chemical Industries, Ltd.) and 0.7% of 5% NaOH aqueous solution were added and reacted at 40 ° C. for 2 hours, White crystals of methylmaleimide were precipitated. After filtration under reduced pressure, it was vacuum dried at room temperature. The crude crystals of hydroxymethylmaleimide thus obtained were recrystallized from ethyl acetate to obtain 22 parts of hydroxymethylmaleimide.

(Examples 1 to 4 and Comparative Examples 1 and 2)
(Introduction of polyoxyethylene chain)
10 parts of each of the crosslinked polymer particles 1 to 6 are dispersed in 75 parts of hydrochloric acid (manufactured by Wako Pure Chemical Industries, Ltd.), 150 parts of dioxane (manufactured by Wako Pure Chemical Industries, Ltd.) are added, and reacted at 80 ° C. for 6 hours. It was. The obtained particles were dispersed / washed in methanol, further washed with ion exchange water and a solvent, and then isolated and dried to obtain polymethacrylic acid-containing carrier particles. From the quantification of carboxylic acid, 86% to 89% of the t-butyl methacrylate component of the copolymer was converted to methacrylic acid in the crosslinked polymer particles 1 to 4. Similarly, from the quantification of carboxylic acid, the copolymer of the crosslinked polymer particles 5 and 6 had 84% to 85% of the t-butyl methacrylate component converted to methacrylic acid.
In 10 parts of the polymethacrylic acid-containing carrier particles thus obtained, 150 parts of polyethylene glycol 200 (manufactured by Wako Pure Chemical Industries, Ltd.), 15 parts of sulfuric acid (manufactured by Wako Pure Chemical Industries, Ltd.), toluene ( 50 parts of Wako Pure Chemical Industries, Ltd.) were mixed and reacted at 120 ° C. for 5 hours while removing generated water out of the system using azeotropy with toluene. The obtained particles are washed with ethanol, acetone, methanol, ion-exchanged water, isolated and dried, and polyoxyethylene chain-introduced crosslinked particles (hydroxyl group-containing polymer particles) 1 to 6 in which polyoxyethylene chains are introduced into the crosslinked polymer particles. Got. The amount of hydroxyl groups in the thus obtained polyoxyethylene chain-introduced crosslinked particles was measured by the following method.

Weigh the particles in a capped test tube, add a certain amount of a pre-prepared acetic anhydride (manufactured by Wako Pure Chemical Industries) solution in pyridine (manufactured by Wako Pure Chemical Industries, Ltd.), and heat at 95 ° C for 24 hours. did.
Further, distilled water was added to hydrolyze acetic anhydride in the test tube, and then centrifuged at 3000 rpm for 5 minutes to separate into particles and supernatant. The particles were further washed with ethanol (manufactured by Wako Pure Chemical Industries, Ltd.) by repeated ultrasonic dispersion and centrifugation, and the supernatant and the washing solution were collected in a conical beaker, and phenolphthalein (manufactured by Wako Pure Chemical Industries, Ltd.) was used as an indicator. The solution was titrated with a 0.1M ethanolic potassium hydroxide solution (Wako Pure Chemical Industries, Ltd.).
A blank experiment without using particles was also performed, and the amount of hydroxyl group (Ammol / g) was calculated from the difference according to the following formula.
A = ((BC) × 0.1 × f) / w
Here, B is the dropping amount (ml) in the blank experiment, C is the dropping amount (ml) of the sample, f is the factor of the potassium hydroxide solution, and w is the weight (g) of the particles.
The results of measuring the hydroxyl groups of the polyoxyethylene chain-introduced crosslinked particles 1 to 6 by the above method are shown in Table 1.

  As can be seen from Table 1, there are particularly few polyoxyethylene chain-introduced crosslinked particles 5 prepared without adding a non-polymerizable solvent and polyoxyethylene chain-introduced crosslinked particles 6 prepared by adding 14% of a non-polymerizable solvent. It never happened.

(Introduction of maleimidyl group)
In each 10 parts of the polyoxyethylene chain-introduced crosslinked particles, 0.8 parts of hydroxymethylmaleimide and 350 parts of toluene were placed and heated and stirred at 60-70 ° C. The temperature was raised and the reaction was carried out under reflux for 7 hours. The obtained particles were dispersed / washed in methanol and further subjected to Soxhlet extraction with ethanol for 8 hours. After filtration, it was washed again with ion-exchanged water and a solvent, and then isolated and dried to obtain maleimidyl group-containing polymer particles 1 to 4 and comparative maleimidyl group-containing polymer particles 5 and 6 of the present invention. The average pore radius and the amount of maleimidyl groups of the thus obtained maleimidyl group-containing polymer particles were measured. The average pore radius was measured by mercury porosimetry using Autopore III9420 (manufactured by MICROMERITICS). The results are shown in Table 2. The amount of maleimidyl groups was measured by the following method.

(Method for quantifying maleimidyl groups in maleimidyl group-containing polymer particles)
A certain amount of particles are weighed into a screw-cap test tube, and a predetermined amount of 2-mercaptoethylamine (manufactured by Tokyo Kasei Co., Ltd.) is added to the reaction solution so that the amount of 2-mercaptoethylamine is excessive, followed by a stirring reaction. I let you.
After centrifuging the particles, the 2-mercaptoamine in the supernatant solution is reacted with 4,4′-dithiodipyridine, and the amount of 2-mercaptoethylamine D in the supernatant solution is determined using the molar absorption coefficient ε = 19800 from the absorbance at 324 nm. mol). As a blank, the 2-mercaptoamine amount E (mol) of a system not containing a sample was measured in the same manner, and a maleimidyl group M (mmol / g) was determined according to the following formula.
M = (ED) / W × 1000

  Table 2 shows the amount of maleimidyl groups of the maleimidyl group-containing polymer particles 1 to 4 of the present invention and the amount of maleimidyl groups of the comparative maleimidyl group-containing polymer particles 5 and 6. Although the same amount of hydroxymethylmaleimide was reacted, the maleimidyl group-containing polymer particles derived from the crosslinked polymer particles according to the present invention had maleimidyl groups that were an order of magnitude higher.

(Reactivity of maleimidyl group-containing carrier particles with terminal SH peptide)
A fixed amount of 100 mg of particles was weighed into a capped microtube, 1 ml of a previously prepared laminin nonapeptide (Calbiochem) solution was added, and the mixture was allowed to react gently for 30 minutes at room temperature. After the particles were centrifuged, the absorbance F at a wavelength of 280 nm of the supernatant solution was measured. As a blank, a wavelength 280 nm absorbance G of a laminin nonapeptide solution of a system not containing a sample was also measured, and a peptide binding rate L% was determined according to the following formula.
L = (1-F / G) × 100
Table 2 shows peptide bond rates of the maleimidyl group-containing polymer particles 1 to 4 of the present invention and the comparative maleimidyl group-containing polymer particles 5 and 6. The maleimidyl group-containing polymer particles of the present invention had high peptide reactivity.

(Scanning electron microscope (SEM) observation of each particle)
The surface of the maleimidyl group-containing polymer particles 1 to 4 of the present invention was observed using a SEM (VE-7800 manufactured by Keyence Corporation) at a magnification of 1000 times. The results are shown in FIGS. A clear porous structure was not observed in the maleimidyl group-containing polymer particles of the present invention. In addition, no cracks, fractured particles, or fused particles were found, and it was found to be mechanically and chemically strong.

2 is a scanning electron micrograph of maleimidyl group-containing polymer particles 1. 2 is a scanning electron micrograph of maleimidyl group-containing polymer particles 2. 2 is a scanning electron micrograph of maleimidyl group-containing polymer particles 3. 4 is a scanning electron micrograph of maleimidyl group-containing polymer particles 4.

Claims (5)

A maleimidyl group-containing polymer particle in which a maleimidyl group represented by the following chemical formula (1) is introduced into a crosslinked polymer particle obtained by copolymerizing a monomer mixture containing a (meth) acrylate, a styrene monomer and a crosslinking agent,
The crosslinked polymer particles are
The monomer mixture is mixed with the non-polymerizable solvent in which the monomer mixture is soluble and the copolymer of the (meth) acrylate and the styrenic monomer is sparingly soluble with respect to the total amount of the monomer mixture. A step of obtaining non-polymerizable solvent-containing particles by copolymerization in a state where the content of the polymerizable solvent is 35% by mass or more;
Removing the non-polymerizable solvent from the non-polymerizable solvent-containing particles;
Maleimidyl group-containing polymer particles obtained through the process.
(In the formula, m and n represent an integer of 1 or more.)   2. The maleimidyl group-containing polymer particle according to claim 1, wherein the non-polymerizable solvent is at least one selected from hydrocarbons having a boiling point of 100 ° C. or higher, long-chain alcohols, long-chain carboxylic acids, and long-chain esters.   The maleidyl group-containing polymer particles according to claim 1 or 2, wherein an average pore radius by a mercury intrusion method is 0.005 to 0.1 µm. A monomer mixture containing a (meth) acrylate, a styrene monomer and a crosslinking agent, a non-polymerizable solvent in which the monomer mixture is soluble and a copolymer of the (meth) acrylate and the styrene monomer is insoluble A step of obtaining non-polymerizable solvent-containing particles by copolymerization in a state where the content of the non-polymerizable solvent is 35% by mass or more based on the total amount with the monomer mixture;
Removing the non-polymerizable solvent from the non-polymerizable solvent-containing particles to obtain crosslinked polymer particles;
Introducing a maleimidyl group represented by the following chemical formula (1) into the crosslinked polymer particles;
The manufacturing method of the maleimidyl group containing polymer particle which has this.
(In the formula, m and n represent an integer of 1 or more.)   The method for producing maleimidyl group-containing polymer particles according to claim 4, wherein the non-polymerizable solvent is at least one selected from hydrocarbons having a boiling point of 100 ° C or higher, long-chain alcohols, long-chain carboxylic acids, and long-chain esters.