JP6777390B2 - Biocompatible medical materials - Google Patents

Description

The present invention relates to biocompatible medical materials. More specifically, the present invention relates to biocompatible medical materials useful, for example, as pharmaceutical carriers, coating agents and the like used when holding a drug and introducing it into the body orally or via blood. The biocompatible medical material of the present invention is expected to be used as, for example, an injectable drug carrier, an orally administered pharmaceutical carrier, and the like.

As a biocompatible material that does not cause a biocompatibility reaction when it enters the living body, it has a synthetic polymer compound having a repeating unit composed of alkoxyalkyl (meth) acrylate as a constituent on the surface and is in contact with biological tissue or body fluid. It consists of a biocompatible medical material used for the application (see, for example, Patent Document 1) and a polymer synthesized by anion polymerization from alkoxyalkyl (meth) acrylate, and is a ratio of weight average molecular weight to number average molecular weight (weight average molecular weight). A biocompatible material having a value of (/ number average molecular weight) of 1.0 to 1.5 and a weight average molecular weight of 40,000 or more (see, for example, Patent Document 2) has been proposed. Both the biocompatible medical material and the biocompatible material can be used for medical instruments and the like that come into contact with blood or biological tissue, but since they do not have sufficient properties to retain the drug, they can hold the drug. Not suitable as a carrier for introduction into the body orally or via blood.

As a composite material (conjugate) containing a bioactive agent, a conjugate in which a bioactive agent or a diagnostic agent is linked to an initiator fragment or a terminal group of a polymer arm has been proposed (see, for example, Patent Document 3). .. However, since a large amount of monomers such as styrene, which are not considered to be suitable for biocompatibility, are used in the biocompatible monomers constituting the conjugate, it is preferable in terms of safety to the human body. However, since the conjugate has a large molecular weight, it has a drawback of being inferior in excretion to the outside of the body.

Japanese Unexamined Patent Publication No. 04-152952 Japanese Unexamined Patent Publication No. 2003-0127223 Special Table 2013-534931

The present invention has been made in view of the above-mentioned prior art, and an object of the present invention is to provide a biocompatible medical material having excellent sustained release of a drug, retention of particle shape, hemolytic resistance and extracorporeal release. And.

The present invention
(1) A biocompatible medical material obtained by polymerizing a biocompatible monomer, having a weight average molecular weight of 1000 to 90000, and containing a biocompatible polymer having a functional group at at least one molecular terminal. there are, alkoxyalkyl (meth) comprising acrylate is polymerized polymer is used as the biocompatible polymer, the functional groups at the molecular ends of the polymer has the formula: -COOM (M is a hydrogen atom or an alkali metal atom group represented by the illustrated), a hydroxyl group, an allyl group, an epoxy group, an aldehyde group, -NH ₂ group, CONH- group or -SH Motodea Ru biocompatible medical material, and (2) to (1) The above-mentioned biocompatible polymer is polymerized with a hydroxyl group-containing (meth) acrylate, and a biocompatible polymer having a weight average molecular weight of 1000 to 90000 and having a functional group at at least one of the molecular ends is bonded. Concerning biocompatible medical materials.

According to the present invention, there is provided a biocompatible medical material having excellent drug sustained release property, particle shape retention property, hemolytic resistance and extracorporeal release property.

As described above, the biocompatible medical material of the present invention is made by polymerizing a biocompatible monomer, has a weight average molecular weight of 1000 to 90000, and has a functional group at at least one molecular terminal. Contains a sex polymer.

Examples of the biocompatible monomer include polyoxyalkylene group-containing monomers, hydroxyl group-containing (meth) acrylates, alkoxyalkyl (meth) acrylates, vinyl monomers, alkylene oxides, alkoxypolyoxyalkylene glycols, cyclic compounds, and amino acids. , Asparagic acid, glutamate, etc., but the present invention is not limited to these examples. These biocompatible monomers may be used alone or in combination of two or more.

In the present invention, "(meth) acrylate" means acrylate or methacrylate, and acrylate and methacrylate may be used alone or in combination. In addition, "(meth) acrylic" means acrylic or methacrylic.

Examples of the polyoxyalkylene group-containing unsaturated monomer include the formula (I):

(In the formula, R ¹ , R ² and R ³ are independently hydrogen atoms or methyl groups, R ⁴ is an alkylene group having 2 to 18 carbon atoms, and R ⁵ is a hydrogen atom or 1 to 20 carbon atoms. The hydrocarbon group X is a divalent alkylene group having 1 to 5 carbon atoms, and when the -CO- group or R ¹ R ³ C = CR ^2- group is a vinyl group, it is directly bonded, and m is-(R ^4). O) -The average number of moles of groups added, indicating the number from 1 to 300)
Examples thereof include a polyoxyalkylene group-containing unsaturated monomer represented by.

When two or more kinds of oxyalkylene groups represented by the formula: -R ⁴ O- are present in one molecule, the oxyalkylene group may be random, blocked or alternate.

In formula (I), R ⁵ is a hydrogen atom or a hydrocarbon group having 1 to 20 carbon atoms. Among R ⁵ , hydrogen atoms and hydrocarbon groups having 1 to 20 carbon atoms are preferable, hydrocarbon groups having 1 to 10 carbon atoms are more preferable, hydrocarbon groups having 1 to 3 carbon atoms are even more preferable, and hydrocarbon groups having 1 to 3 carbon atoms are even more preferable. One or two hydrocarbon groups are more preferred. Among the hydrocarbon groups, a saturated alkyl group and an unsaturated alkyl group are preferable, a saturated alkyl group having 1 to 20 carbon atoms is more preferable, a saturated alkyl group having 1 to 10 carbon atoms is further preferable, and a saturated alkyl group having 1 to 10 carbon atoms is more preferable. Saturated alkyl groups are even more preferred.

In formula (I), wherein: the oxyalkylene group represented by -R ⁴ O-is an oxyalkylene group having 2 to 18 carbon atoms. Examples of the oxyalkylene group include an oxyethylene group, an oxypropylene group, an oxybutylene group, an oxyisobutylene group, an oxy-1-butene group, an oxy-2-butene group, and the like. It is not limited to. Among these oxyalkylene groups, an oxyalkylene group having 2 to 8 carbon atoms is preferable, and an oxyalkylene group having 2 to 4 carbon atoms such as an oxyethylene group, an oxypropylene group and an oxybutylene group is more preferable, and an oxyethylene group. Is even more preferable.

In formula (I), m, wherein: the average number of moles of the oxyalkylene group represented by -R ⁴ O-. The average number of moles added means the average number of moles of oxyalkylene groups in 1 mole of the unsaturated monomer containing a polyoxyalkylene group. The lower limit of m is preferably 2 or more, more preferably 4 or more, and further preferably 8 or more. The upper limit of m is preferably 100 or less, more preferably 50 or less.

X is a direct bond when the divalent alkylene group having 1 to 5 carbon atoms, the −CO− group, or the R ¹ R ³ C = CR ² − group is a vinyl group. Of these groups, the -CO- group is preferred.

Examples of the polyoxyalkylene group-containing unsaturated monomer include unsaturated alcohol polyalkylene glycol adducts, polyalkylene glycol ester monomers, and (alkoxy) polyalkylene glycol monomaleic acid esters.

An unsaturated alcohol polyalkylene glycol adduct is a compound in which a polyalkylene glycol chain is added to an alcohol having an unsaturated group. Examples of the unsaturated alcohol polyalkylene glycol adduct include polyethylene glycol monovinyl ether, polyethylene glycol monoallyl ether, polyethylene glycol mono (2-methyl-2-propenyl) ether, polyethylene glycol mono (2-butenyl) ether, and polyethylene glycol. Mono (3-methyl-3-butenyl) ether, polyethylene glycol mono (3-methyl-2-butenyl) ether, polyethylene glycol mono (2-methyl-3-butenyl) ether, polyethylene glycol mono (2-methyl-2-) Butenyl) ether, polyethylene glycol mono (1,1-dimethyl-2-propenyl) ether, polyethylene glycol glycol mono (3-methyl-3-butenyl) ether, methoxypolyethylene glycol mono (3-methyl-3-butenyl) ether, etc. However, the present invention is not limited to such examples.

The polyalkylene glycol ester-based monomer is a monomer in which an unsaturated group and a polyalkylene glycol chain are bonded via an ester bond.

As the polyalkylene glycol ester-based monomer, for example, an esterified product of an alkoxypolyalkylene glycol in which 1 to 300 mol of an oxyalkylene group having 2 to 18 carbon atoms is added to an alcohol and (meth) acrylic acid is preferable. Among the alkoxypolyalkylene glycols, those containing an oxyethylene group as a main component are preferable. Examples of the alcohol include methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, 1-pentanol, 2-pentanol, 3-pentanol, 1-hexanol, 2-hexanol. 3-Hexanol, octanol, 2-ethyl-1-hexanol, nonyl alcohol, lauryl alcohol, cetyl alcohol, stearyl alcohol and other aliphatic alcohols with 1 to 30 carbon atoms, cyclohexanol and other alicyclic alcohols with 3 to 30 carbon atoms Examples thereof include alcohols, (meth) allyl alcohols, 3-butene-1-ol, 3-methyl-3-buten-1-ol and other unsaturated alcohols having 3 to 30 carbon atoms, which are used alone. Also, two or more types may be used in combination. Examples of the esterified product include methoxypolyethylene glycol mono (meth) acrylate, methoxy (polyethylene glycol polypropylene glycol) mono (meth) acrylate, methoxy (polyethylene glycol polybutylene glycol) mono (meth) acrylate, and methoxy (polyethylene glycol polypropylene glycol). Examples thereof include polybutylene glycol) mono (meth) acrylate, and these may be used alone or in combination of two or more.

Among the polyalkylene glycol ester-based monomers, (alkoxy) polyalkylene glycol mono (meth) acrylates such as methoxypolyethylene glycol monomethacrylate are preferable.

Examples of the hydroxyl group-containing (meth) acrylate include hydroxyalkyl (meth) acrylate, glycerin acrylate, and glycerin methacrylate having 2 to 4 carbon atoms in the hydroxyalkyl group such as 2-hydroxyethyl acrylate and 2-hydroxyethyl methacrylate. However, the present invention is not limited to such an example. These hydroxyl group-containing (meth) acrylates may be used alone or in combination of two or more.

Examples of the alkoxyalkyl (meth) acrylate include methoxymethyl (meth) acrylate, methoxyethyl (meth) acrylate, methoxypropyl (meth) acrylate, ethoxymethyl (meth) acrylate, ethoxyethyl (meth) acrylate, and ethoxypropyl (meth) acrylate. Examples thereof include alkoxyalkyl (meth) acrylates in which an alkoxy group such as a meta) acrylate has 1 to 4 carbon atoms and an alkyl group having 1 to 4 carbon atoms, but the present invention is limited to such examples. It's not something. These alkoxyalkyl (meth) acrylates may be used alone or in combination of two or more.

Examples of the vinyl monomer include 2- (meth) acryloyloxymethylphosphorylcholine, 2- (meth) acryloyloxyethylphosphorylcholine, tetrahydrofurfuryl (meth) acrylate, isopropylacrylamide, vinyl alcohol, vinylformamide, vinylisobutylacrylamide, and the like. Meta) Acrylamide, dimethylacrylamide, vinylacetamide, N-vinylpyrrolidone and the like, but the present invention is not limited to these examples. These vinyl monomers may be used alone or in combination of two or more.

Examples of the alkylene oxide include alkylene oxides having 2 to 4 carbon atoms such as ethylene oxide and propylene oxide, but the present invention is not limited to these examples. These alkylene oxides may be used alone or in combination of two or more.

Examples of the alkoxypolyoxyalkylene glycol include an alkoxy group having 1 to 4 carbon atoms such as polyethylene glycol, polypropylene glycol, methoxypolyethylene glycol, ethoxypolyethylene glycol, methoxypolypropylene glycol, and ethoxypolypropylene glycol, and an oxy having 1 to 4 carbon atoms. Examples thereof include alkoxypolyoxyalkylene glycol having an alkylene group and having 2 to 30 addition moles of the oxyalkylene group, but the present invention is not limited to such examples. These alkoxypolyoxyalkylene glycols may be used alone or in combination of two or more.

Examples of the cyclic compound include lactides such as L-lactide, lactones such as ε-caprolactone, trimethylcarbonate, cyclic amino acids, morpholine-2,5-dione, and the like. It is not limited to. Each of these cyclic compounds may be used alone, or two or more kinds may be used in combination.

Among the biocompatible monomers, a polyoxyalkylene group-containing unsaturated monomer, a hydroxyalkyl (meth) acrylate having a hydroxyalkyl group having 2 to 4 carbon atoms, and an alkoxy group having 1 to 4 carbon atoms are used. Alkoxyalkyl acrylate having an alkyl group having 1 to 4 carbon atoms, an alkoxy group having 1 to 4 carbon atoms and an oxyalkylene group having 1 to 4 carbon atoms, and an addition molar number of 2 to 30 oxyalkylene groups. Alkoxypolyoxyalkylene glycol and tetrahydrofurfuryl (meth) acrylate are preferable, and polyoxyalkylene group-containing unsaturated monomer, methoxyethyl acrylate, 2-hydroxyethyl (meth) acrylate, N-vinylpyrrolidone, 2-(. More preferred are acryloyloxyethyl phosphorylcholine, methoxypolykoxyglycol and tetrahydrofurfuryl (meth) acrylate.

The biocompatible monomer can be used in combination with another monomer copolymerizable with the biocompatible monomer as long as the object of the present invention is not impaired. Examples of the other monomer include aziridines, (meth) acrylic acid, methyl (meth) acrylate, ethyl (meth) acrylate, propyl (meth) acrylate, n-butyl (meth) acrylate, and isobutyl (meth). Acrylate, tert-butyl (meth) acrylate, 2-ethylhexyl (meth) acrylate, diaminomethyl (meth) acrylate, diaminoethyl (meth) acrylate, dimethylamino (meth) acrylate, diethylamino (meth) acrylate, glycidyl (meth) acrylate , Acrylate, etc., but the present invention is not limited to such examples. These other monomers may be used alone or in combination of two or more.

Examples of the method for polymerizing the biocompatible monomer include a living radical polymerization method represented by a radical polymerization method, an atom transfer radical polymerization method, and a reversible addition cleavage chain transfer polymerization method, an ion polymerization method, and a ring-opening weight. Legal, radical polymerization methods, polycondensation methods and the like can be mentioned, but the present invention is not limited to these examples.

A solvent may be used when polymerizing the biocompatible monomer. Examples of the solvent include aromatic solvents such as benzene, toluene and xylene; alcohol solvents such as methanol, ethanol, isopropanol, n-butanol and tert-butyl alcohol; halogen atom-containing solvents such as dichloroethane and dichloroethane; propylene glycol. Ether-based solvents such as methyl ether, dipropylene glycol methyl ether, ethyl cellosolve, butyl cellosolve; ester solvents such as ethyl acetate, butyl acetate, cellosolve acetate; ketone solvents such as acetone, methyl ethyl ketone, methyl isobutyl ketone, diacetone alcohol; Organic solvents such as amide solvents such as dimethylformamide can be mentioned, but the present invention is not limited to these examples. Each of these solvents may be used alone, or two or more kinds may be used in combination. The amount of the solvent may be appropriately determined in consideration of the polymerization conditions, the composition of the biocompatible monomer, the concentration of the obtained polymer and the like.

When polymerizing the biocompatible monomer, a chain transfer agent can be used to adjust the molecular weight of the biocompatible polymer. The chain transfer agent includes those corresponding to the functional group-introduced compound, and the chain transfer agent corresponding to the functional group-introduced compound may be used as one or both of the chain transfer agent and the functional group-introduced compound. it can.

Examples of the chain transfer agent include alkali metal salts of thioacetate such as sodium thioacetate and potassium thioacetate, cysteine, cysteamine, mercaptoethanol, thioglycerol, thioglycolic acid, mercaptopropionic acid, 2-mercaptopropionic acid, and 3-mercapto. Hydrophilic thiol-based chain transfer agents such as propionic acid, thioacetic acid, thioannic acid, 2-mercaptoethanesulfonic acid, their sodium salts and potassium salts; primary alcohols such as 2-aminopropan-1-ol, isopropanol and the like. Secondary alcohols, phosphite, hypophosphite and salts thereof (eg, sodium hypophosphite, potassium hypophosphite, etc.), sulfites, hydrogen sulfites, dithionic acids, metabisulfites and salts thereof Non-thiol chain transfer agents such as (eg, sodium sulfite, sodium hydrogen sulfite, sodium dithionate, sodium metabisulfite, potassium sulfite, potassium hydrogen sulfite, potassium dithionate, potassium metabisulfite, etc.); Butane thiol, octane thiol, decane thiol, dodecane thiol, hexadecane thiol, octadecane thiol, cyclohexyl mercaptan, thiophenol, octyl thioglycolate, octyl 2-mercaptopropionate, octyl 3-mercaptopropionate, 2-ethylhexyl mercaptopropionate , Octanoic acid 2-mercaptoethyl ester, 1,8-dimercapto-3,6-dioxaoctane, decantrythiol, dodecyl mercaptan and other hydrophobic thiol-based chain transfer agents. It is not limited to only. Each of these chain transfer agents may be used alone, or two or more of them may be used in combination.

The amount of the chain transfer agent may be appropriately set according to the type of the biocompatible monomer, the polymerization conditions such as the polymerization temperature, the target molecular weight of the biocompatible polymer, etc., and is not particularly limited, but is a weight average. When a polymer having a molecular weight of several thousand to tens of thousands is obtained, it is preferably 0.1 to 20 parts by mass, and 0.5 to 15 parts by mass per 100 parts by mass of the biocompatible monomer. Is more preferable.

When polymerizing the biocompatible monomer, a polymerization initiator can be used. In addition, the polymerization initiator includes those corresponding to the functional group-introduced compound, and the polymerization initiator corresponding to the functional group-introduced compound may be used as one or both of the polymerization initiator and the functional group-introduced compound. it can.

Examples of the polymerization initiator include azoisobutyronitrile, 4,4'-azobis (4-cyanopentanoic acid), and 2,2'-azobis [2-methyl-N- [1,1-bis (hydroxy)). Methyl) -2-hydroxyethyl] propionamide], 2,2'-azobis [N- (2-hydroxyethyl) -2-methoxypropaneamide], 2,2'-azobis (2-methyl-2-propenylpropane) Amid), 2,2'-bis (2-imidazolin-2-yl) [2,2'-azobispropane] dihydrochloride, 2,2'-azobis (propane-2-carbamomidin) dihydrochloride, 2,2'-azobis [N- (2-carboxyethyl) -2-methylpropion amidine], 2,2'-azobis [2- [1- (2-hydroxyethyl) -2-imidazolin-2-yl] Propane dihydrochloride, 2,2'-azobis (2,4-dimethylvaleronitrile), 2,2'-azobis (2-methylbutyronitrile), tert-butylperoxy-2-ethylhexanoate, 2 , 2'-Azobisisobutyronitrile, benzoyl peroxide, ditert-butyl peroxide, cyclohexanone peroxide, acetylacetone peroxide and other radical polymerization initiators; bromomethylbenzene, 1- (bromomethyl) -4-methylbenzene , 2-Bromoisobutyrate ethyl, 2-bromoisobutyrate hydroxyethyl, bis [2- (2'-bromoisobutyryloxy) ethyl] disulfide, 2-bromoisobutyrate 10-undecenyl, 4- (1-bromoethyl) benzoic acid, etc. However, the present invention is not limited to such examples. Each of these polymerization initiators may be used alone, or two or more kinds thereof may be used in combination.

When a living polymerization initiator is used as the polymerization initiator, the polymer is prepared by reacting a functional group-containing compound with a halogen atom existing at the terminal of the polymer prepared by using the living polymerization initiator. A functional group may be introduced. Examples of the functional group-containing compound include amine compounds such as ethylenediamine and propyldiamine, dithiol compounds such as ethanedithiol, propanedithiol and hexadecanedithiol, allyl mercaptoethanol, cysteine, cysteamine, mercaptoethanol, thioglycerol and thioglycol. Examples thereof include acids, mercaptopropionic acid, 2-mercaptopropionic acid, 3-mercaptopropionic acid, thioacetic acid, thioannic acid, 2-mercaptoethanesulfonic acid, and thiol compounds such as sodium salts and potassium salts thereof. Is not limited to such examples.

The amount of the polymerization initiator may be appropriately set according to the desired physical properties of the obtained polymer, but is usually 0.001 to 20 parts by mass, more preferably 0.001 to 20 parts by mass per 100 parts by mass of the biocompatible monomer. It is preferably 0.005 to 10 parts by mass.

In order to obtain a biocompatible polymer having a functional group, a functional group-containing compound for introducing a functional group into the biocompatible polymer can be used. As the functional group, an anionic functional group, a cationic functional group, a nonionic functional group and an amphoteric functional group are preferable. The functional group is preferably a reactive functional group from the viewpoint of modifying a drug or the like. Suitable functional groups include, for example, a group represented by the formula: -COMM (M represents a hydrogen atom or an alkali metal atom), a hydroxyl group, an allyl group, an epoxy group, an aldehyde group, -NH ₂ groups, a CONH- group, and the like. -SH groups and the like can be mentioned, but the present invention is not limited to such examples. Examples of the M include alkali metal atoms such as sodium atom and potassium atom. The number of functional groups in the biocompatible polymer having a functional group is not particularly limited, but is usually preferably 1 to 6.

Examples of the functional group-containing compound for introducing a functional group into the biocompatible polymer include alkali metal salts of thioacetate such as sodium thioacetate and potassium thioacetate, cysteine, cysteamine, mercaptoethanol, thioglycerol, and thioglycolic acid. , Mercaptopropionic acid, 2-mercaptopropionic acid, 3-mercaptopropionic acid, thioacetic acid, thioannic acid, 2-mercaptoethanesulfonic acid, their sodium salts, potassium salts and other thiol-based chain transfer agents; 4,4'- Azobis (4-cyanopentanoic acid), 2,2'-azobis [2-methyl-N- [1,1-bis (hydroxymethyl) -2-hydroxyethyl] propionamide], 2,2'-azobis [N -(2-Hydroxyethyl) -2-methoxypropanamide], 2,2'-azobis (2-methyl-2-propenylpropanamide), 2,2'-bis (2-imidazolin-2-yl) [2 , 2'-azobispropane] dihydrochloride, 2,2'-azobis (propane-2-carbamomidin) dihydrochloride, 2,2'-azobis [N- (2-carboxyethyl) -2-methylpropion Amidin], 2,2'-azobis [2- [1- (2-hydroxyethyl) -2-imidazolin-2-yl] propan dihydrochloride, cyclohexanone peroxide, acetylacetone peroxide and other functional groups were introduced. Examples thereof include a polymerization initiator, but the present invention is not limited to these examples. Each of these functional group-containing compounds may be used alone, or two or more kinds may be used in combination.

The amount of the functional group-containing compound for introducing a functional group into the biocompatible polymer, the type of biocompatible monomers, polymerization conditions such as polymerization temperature, depending on the molecular weight of the biocompatible polymer to the target It may be appropriately set, but is not particularly limited, but is 0.1 to 20 parts by mass per 100 parts by mass of the biocompatible monomer when a polymer having a weight average molecular weight of several thousand to tens of thousands is obtained. It is preferably 0.5 to 15 parts by mass, and more preferably 0.5 to 15 parts by mass.

The polymerization conditions for polymerizing the biocompatible monomer may be appropriately set according to the polymerization method, and are not particularly limited. The polymerization temperature is preferably room temperature to 200 ° C., more preferably 40 to 140 ° C. Further, the atmosphere for polymerizing the biocompatible monomer is preferably an inert gas such as nitrogen gas or argon gas. The reaction time may be appropriately set so that the polymerization reaction of the biocompatible monomer is completed.

As a method of introducing a functional group into the terminal of a biocompatible monomer or a biocompatible polymer, for example,
(1) A method for polymerizing a biocompatible monomer in the presence of a polymerization initiator into which the functional group has been introduced as a polymerization initiator.
(2) A method for polymerizing a biocompatible monomer in the presence of a chain transfer agent into which the functional group has been introduced as a chain transfer agent.
(3) When the biocompatible polymer has an allyl group at the terminal, the allyl group of the biocompatible polymer is boronized, and then the boronized biocompatible polymer and the functional group-containing compound are combined. A method for introducing a functional group of the functional group-containing compound by reacting,
(4) When the biocompatible polymer has a hydroxyl group at the terminal, a method of introducing a thiol group as a functional group by reacting the hydroxyl group with a trilling agent such as tosyl chloride and then acetylating the polymer.
(5) Examples thereof include a method of reacting a halogen atom existing at the terminal of a biocompatible polymer with a functional group-containing compound, but the present invention is not limited to these examples.

By polymerizing the biocompatible monomer as described above, a biocompatible polymer can be obtained. The resulting biocompatible polymer has a functional group at its end. The functional group may be present only at one end of the biocompatible polymer, or may be present at both ends. When the biocompatible polymers are bonded to each other, a biocompatible polymer having functional groups at both ends is used as the biocompatible polymer. The biocompatible polymer has a functional group contained in the compound for introducing a functional group into the biocompatible polymer . Suitable functional groups of the biocompatible polymer include, for example, a group represented by the formula: -COMM (M is the same as above), a hydroxyl group, an allyl group, an epoxy group, an aldehyde group, -NH ₂ groups, and a CONH- group. , -SH groups and the like, but the present invention is not limited to such examples.

The biocompatible polymer may be a biocompatible block copolymer obtained by binding the same type or different types of biocompatible polymers to each other.

As a method for binding the biocompatible polymers to each other, for example, as shown in the following production examples, after the functional groups of the biocompatible polymers are chemically reacted, amidation, esterification, etherification, and carbonate are used. Examples thereof include a method of binding biocompatible polymers to each other by esterification, dehydration condensation, etc., but the present invention is not limited to such examples. Examples of the functional group that binds the biocompatible polymers to each other include an amide bond, an ester bond, a carbonate bond, an ether bond, and an acid anhydride bond, but the present invention is limited to these examples only. is not.

The weight average molecular weight of the biocompatible polymer is 1000 or more, preferably 2000 or more, more preferably 3000 or more, from the viewpoint of improving the sustained release property, particle shape retention, hemolytic resistance and extracorporeal release property of the drug. From the viewpoint of improving the in vitro release property, it is 90,000 or less, preferably 50,000 or less, more preferably 30,000 or less, and further preferably 25,000 or less.

The weight average molecular weight of the biocompatible polymer means a value when measured based on the method described in the following Examples.

The molecular weight distribution of the biocompatible polymer (value of [polymerization average molecular weight / number average molecular weight]) is preferably 1 to 2, more preferably 1 to 1.5, and even more preferably 1 to 2 from the viewpoint of improving hemolytic resistance. It is 1-1.3.

The biocompatible polymer can be used as particles. The average particle size of the particles is preferably 10 nm or more, more preferably 20 nm or more from the viewpoint of not acting on normal cells, and preferably 1000 nm or less, more preferably 500 nm or less from the viewpoint of acting on lesion cells. , More preferably 200 nm or less. The average particle size of the particles is a value measured based on the method described in the following examples.

If necessary, the functional groups of the biocompatible polymer may be modified with proteins, monosaccharides, polysaccharides, sugar chains, antibodies, nucleic acids, drug substances and the like. These proteins, monosaccharides, polysaccharides, sugar chains, antibodies, nucleic acids, drug substances, etc. may be directly bound to the functional groups of the biocompatible polymers and biocompatible copolymers, or may be directly bound to the functional groups of the biocompatible polymers, via a linker. May be combined.

The biocompatible material of the present invention contains a biocompatible polymer. The biocompatible medical material of the present invention may contain additives as long as the object of the present invention is not impaired. Additives include, for example, water, physiological saline, pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, sodium carboxymethyl cellulose salt, sodium polyacrylate, sodium alginate, water-soluble dextran. , Carboxymethyl starch sodium, pectin, methyl cellulose, ethyl cellulose, xanthan gum, arabic rubber, casein, gelatin, agar, diglycerin, propylene glycol, polyethylene glycol, vaseline, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, Examples thereof include lactose, phosphate-buffered physiological saline, biodegradable polymers, serum-free media, surfactants that are acceptable as pharmaceutical additives, and physiological pH buffers that are acceptable in vivo. , It is not limited to such an example. Each of these additives may be used alone, or two or more kinds may be used in combination.

The biocompatible medical material of the present invention can be used, for example, as a carrier for holding a medicine or the like. As a method of holding a medicine or the like with the biocompatible medical material of the present invention, for example, a carrier is formed by binding the medicine or the like to a functional group of the biocompatible polymer used in the biocompatible medical material. A method of combining biocompatible medical materials with pharmaceuticals, a method of mixing biocompatible medical materials and pharmaceuticals so as to have a uniform composition, a method of coating particles of pharmaceuticals with biocompatible medical materials, lipids and A method of encapsulating a mixture with a biocompatible medical material into particles and encapsulating a drug or the like inside the obtained particles, or by coating a particle containing a drug or the like with a liposome with a biocompatible medical material. A method of encapsulating the particles inside the outer skin of a compatible medical material, by coating the particles of a mixture of a drug and a liposome with the biocompatible medical material, the inside of the outer skin of the biocompatible medical material Examples thereof include a method of encapsulating the particles and a method of encapsulating the drug by micellarizing the drug or the like with a biocompatible medical material, but the present invention is not limited to such examples.

The liposome was prepared by, for example, dissolving the lipid in a solvent such as tert-butyl alcohol and then freeze-drying, or adding a solution in which the drug was dissolved to the lipid to swell the lipid and disperse it with ultrasonic waves. Later, it can be prepared by a method of adding polyethylene glycol-phosphatidylethanolamine or the like to the obtained dispersion.

The liposomes are preferably cationized with a cationizing agent. Liposomes can be obtained by, for example, dissolving hydrided soybean lecithin, cholesterol, 3,5-dipentadecyloxybenzamidine hydrochloride or the like in a solvent such as tert-butyl alcohol and freezing the obtained lipid mixed solution. be able to. The lipids that make up liposomes are stable in vivo. Examples of the lipid include lipids derived from biomaterials, phospholipids or derivatives thereof, lipids other than phospholipids or derivatives thereof, and the like. Examples of the phospholipid include phospholipids such as phosphatidylcholine (lecithin), phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, and cardiolibin, and phospholipids to which hydrogen is added.

In the above example, liposomes have been mentioned, but instead of liposomes, for example, emulsions, nanoparticles, microparticles, polymer compounds and the like can be used.

As the drug, a biologically or pharmacologically active drug can be used. Drugs include, for example, antitumor agents, anticancer agents, antibiotics, antiviral agents, anticancer effect enhancers, immunopotentiators, immunomodulators, immunorecovery agents, radiation sensitizers, radiation protective agents, antihistamines, anti-inflammatory agents. Agents, congestion removers, antifungal agents, anti-arteritis agents, anti-asthma agents, angiogenesis inhibitors, enzyme agents, antioxidants, hormones, angiotensin converting enzyme inhibitors, smooth muscle cell growth agents, smooth muscle cell migration Inhibitors, platelet aggregation inhibitors, chemical mediator release inhibitors, vascular endothelial cell growth promoters, vascular endothelial cell growth inhibitor, interferon, interleukin, colony stimulating factor, cytokines, tumor necrosis factor, granulocyte macrophage colony Stimulating factor, granulocyte colony stimulating factor, macrophage colony stimulating factor, stem cell factor, β-type transforming growth factor, hepatocellular growth factor, vascular endothelial growth factor, erythropoetin, vaccine, protein, mucoprotein, peptide, polysaccharide, lipo Examples include, but are not limited to, polysaccharides, sugar chains, antisense, ribozymes, decoys, nucleic acids, antibodies and the like. Each of these agents may be used alone or in combination of two or more.

Although the biocompatible medical material is obtained as described above, the biocompatible polymer or the biocompatible copolymer used for the biocompatible medical material may be crosslinked. Examples of the method for cross-linking the polymer include a chemical cross-linking method and a physical cross-linking method. Examples of the chemical cross-linking method include epoxy compounds, oxidized starch, glutaaldehyde, formaldehyde, dimethyl suberiminoate, carbodiimide, succinimidyl compounds, diisosianato compounds, acyl azide, reuterin, tris (hydroxymethyl) phosphine, copper ascorbate, and glucose. Methods such as cross-linking the polymer with a chemical cross-linking agent such as lysine or photooxidant, heat dehydration treatment, irradiation with ultraviolet rays, irradiation with electron beams, irradiation with gamma rays, etc. Can be mentioned. The physical cross-linking method includes, for example, a method of cross-linking the polymer with a salt, a method of cross-linking the polymer by electrostatic interaction, a method of cross-linking the polymer by hydrogen bonding, and a method of cross-linking the polymer by hydrophobic interaction. There is a method of cross-linking the polymer. As the cross-linking method, only one type may be used, or two or more types may be used in combination.

The subject to which the drug is administered includes mammals such as humans, monkeys, mice, and domestic animals, but the present invention is not limited to such examples.

When the drug is administered by injection, the drug can be injected into the body by, for example, intravenous injection such as instillation, intramuscular injection, intraperitoneal injection, subcutaneous injection, intradermal injection, intratumoral injection, or the like.

The amount of the medicine to be retained in the biocompatible medical material of the present invention cannot be unconditionally determined because it varies depending on the subject to which the medicine is administered, the type of the medicine, and the like, but usually, the biocompatible medicine of the present invention is used. It is preferably about 1 μg to 50 g per 100 g of the polymer (solid content) contained in the material.

As described above, the biocompatible medical material of the present invention is excellent in sustained release of the drug, retention of particle shape, hemolytic resistance and extracorporeal release, so that the drug can be retained orally or orally. It is expected to be used as a medicinal carrier used for introduction into the body via blood, for example, an injectable medicinal carrier, an orally administered medicinal carrier, and the like.

Next, the present invention will be described in more detail based on Examples, but the present invention is not limited to such Examples.

Manufacturing example 1
180 g of toluene was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the temperature of the toluene in the reaction vessel was raised to 90 ° C. to 20 mL / The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel at a flow rate of min for 30 minutes.

Next, a mixed solution of 100 g of methoxyethyl acrylate, 1.6 g of potassium thioacetate, 0.386 g of azobisisobutyronitrile and 20 g of toluene was continuously added dropwise to a reaction vessel kept at 90 ° C. over 2 hours. While maintaining the temperature of the contents of the reaction vessel at 90 ° C., 0.1 g of azobisisobutyronitrile was added into the reaction vessel 3 hours and 4 hours after the start of dropping the mixed solution, respectively, and then the contents of the reaction vessel. A polymer solution was obtained by maintaining the temperature of the product at 90 ° C. for 2 hours.

100 g of methanol was added to the polymer solution obtained above, and 15 g of a methanol solution containing 0.1 g of sodium hydroxide was added to the polymer solution, and then the polymer solution was stirred at 65 ° C. for 4 hours. ..

Next, 0.2 g of acetic acid was added to the polymer solution to obtain a polymer solution containing a polymethoxyethyl acrylate having a thiol group at the terminal [hereinafter referred to as polymer (1)]. The solid content in the polymer solution was 22.5% by mass, and the weight average molecular weight of the polymer (1) was 5000.

In each Example and each Comparative Example, the weight average molecular weight of the polymer was measured by gel permeation chromatography under the following measurement conditions.
[Measurement conditions for weight average molecular weight of polymer]
-Measuring equipment: Tosoh Corporation, product number: HLC-8320GPC
-Polymer weight column: manufactured by Toso Co., Ltd., product number: TSKgel G4000HHR connected in series-Eluent: Dimethylformamide-Standard material for calibration curve: Polyethylene glycol-Preparation of solution for measurement: Heavy on eluent (dimethylformamide) A solution having a polymer concentration of 0.3% by mass was prepared by dissolving the coalescence, and the solution was used after filtering with a filter.

Manufacturing example 2
180 g of isopropanol was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the isopropanol in the reaction vessel was refluxed at a flow rate of 20 mL / min. The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel for 30 minutes. Then, a mixed solution of 100 g of methoxyethyl acrylate, 0.4 g of 4,4'-azobis (4-cyanopentanoic acid) and 20 g of isopropanol was continuously added dropwise into the reaction vessel over 2 hours.

Next, while maintaining the reflux state, 0.1 g of 4,4'-azobis (4-cyanopentanoic acid) is added 3 hours and 4 hours after the start of dropping the mixed solution, respectively, and the reflux state is maintained for 2 hours. As a result, a polymer solution containing polymethoxyethyl acrylate having a carboxyl group at the terminal [hereinafter referred to as polymer (2)] was obtained. The solid content in the polymer solution was 33.0% by mass, and the weight average molecular weight of the polymer (2) was 5500.

Manufacturing example 3
After adding a mixture of 200 mg of 4- (1-bromoethyl) benzoic acid, 5.2 g of methoxyethyl acrylate and 5 g of n-butanol into a 100 mL Schlenk tube, the internal space of the Schlenk tube was replaced with nitrogen gas. A reaction solution was obtained by adding 400 mg of a bipyridine ligand and 147 mg of copper bromide into the Schlenk tube and stirring for 3 hours while maintaining the temperature of the contents of the Schlenk tube at 80 ° C. The resulting reaction solution was purified by passing it through a silica column.

Next, 0.15 g of sodium hydroxide and 0.2 g of cysteamine were added to the purified reaction solution, and the reaction solution was stirred at a temperature of 80 ° C. for 4 hours to have a carboxyl group at one end and the other. A polymer solution containing a polymethoxyethyl acrylate having an amino group at the terminal of the above [hereinafter referred to as polymer (3)] was obtained. The solid content in the polymer solution was 45% by mass, and the weight average molecular weight of the polymer (3) was 4900.

Manufacturing example 4
After adding 200 mg of 4- (1-bromoethyl) benzoic acid, 5.2 g of 2-hydroxyethyl methacrylate, and 5 g of a mixed solvent consisting of 50% by mass of methanol and 50% by mass of water into a 100 mL Schlenk tube, the inside of the Schlenk tube. The space was replaced with nitrogen gas. A reaction solution was obtained by adding 400 mg of a bipyridine ligand and 147 mg of copper bromide into the Schlenk tube and stirring for 3 hours while maintaining the temperature of the contents of the Schlenk tube at 80 ° C. The resulting reaction solution was purified by passing it through a silica column.

Next, 1 g of a 1N aqueous sodium hydroxide solution and 1.0 g of 1,3-propanedithiol were added to the purified reaction solution, and the reaction solution was stirred at a temperature of 80 ° C. for 4 hours to cause a carboxyl group at one end. A polymer solution containing polyhydroxyethyl methacrylate [hereinafter referred to as polymer (4)] having a thiol group at the other end was obtained. The solid content in the polymer solution was 43% by mass, and the weight average molecular weight of the polymer (4) was 5000.

Production example 5
Add 180 g of isopropanol into a 500 mL reaction vessel equipped with a thermometer, stirrer, dropping funnel, nitrogen gas introduction tube and reflux condenser, and at a flow rate of 20 mL / min while refluxing the contents of the reaction vessel. The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel for 30 minutes.

Next, while maintaining the contents of the reaction vessel in a reflux state, a mixed solution of 100 g of 2-hydroxyethyl methacrylate, 1.6 g of potassium thioacetate, 0.386 g of azobisisobutyronitrile and 20 g of isopropanol was added over 2 hours. It was continuously added dropwise into the reaction vessel. While maintaining the reflux state of the contents of the reaction vessel, 0.1 g of azobisisobutyronitrile is added 3 hours and 4 hours after the start of dropping of the mixed solution, respectively, and the reflux state is further maintained at 90 ° C. for 2 hours. As a result, a polymer solution was obtained.

100 g of methanol was added to the polymer solution obtained above, and 15 g of a methanol solution containing 0.1 g of sodium hydroxide was further added, and then the polymer solution was stirred at 65 ° C. for 4 hours.

Next, 0.2 g of acetic acid was added to the polymer solution to obtain a polymer solution containing polyhydroxyethyl methacrylate having a thiol group at the terminal [hereinafter referred to as polymer (5)]. The solid content in the polymer solution was 23.0% by mass, and the weight average molecular weight of the polymer (5) was 5000.

Production example 6
After adding 200 mg of 4- (1-bromoethyl) benzoic acid, 5.2 g of 2-hydroxyethyl methacrylate, and 5 g of a mixed solvent consisting of 50% by mass of methanol and 50% by mass of water into a 100 mL Schlenk tube, the inside of the Schlenk tube. The space was replaced with nitrogen gas. A reaction solution was obtained by adding 400 mg of a bipyridine ligand and 147 mg of copper bromide into the Schlenk tube and stirring for 3 hours while maintaining the temperature of the contents of the Schlenk tube at 80 ° C. The resulting reaction solution was purified by passing it through a silica column.

Next, 1 g of a 1N aqueous sodium hydroxide solution and 0.2 g of cysteamine were added to the purified reaction solution, the reaction solution was stirred at a temperature of 80 ° C. for 4 hours, and then freeze-dried to cause a carboxyl group at one end. A polymer solution containing polyhydroxyethyl methacrylate having an amino group at the other end [hereinafter referred to as polymer (6)] was obtained. The solid content in the polymer solution was 42% by mass, and the weight average molecular weight of the polymer (6) was 5400.

Production example 7
Add 180 g of isopropanol into a 500 mL reaction vessel equipped with a thermometer, stirrer, dropping funnel, nitrogen gas introduction tube and reflux condenser, and at a flow rate of 20 mL / min while refluxing the contents of the reaction vessel. The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel for 30 minutes.

Next, while maintaining the contents of the reaction vessel in a reflux state, a mixed solution of 100 g of N-vinylpyrrolidone, 1.6 g of potassium thioacetate, 0.386 g of azobisisobutyronitrile and 20 g of isopropanol was reacted over 2 hours. It was continuously dropped into the container. While maintaining the reflux state of the contents of the reaction vessel, 0.1 g of azobisisobutyronitrile is added 3 hours and 4 hours after the start of dropping of the mixed solution, respectively, and the reflux state is further maintained at 90 ° C. for 2 hours. As a result, a polymer solution was obtained.

100 g of methanol was added to the polymer solution obtained above, and 15 g of a methanol solution containing 0.1 g of sodium hydroxide was further added, and then the polymer solution was stirred at 65 ° C. for 4 hours.

Next, 0.2 g of acetic acid was added to the polymer solution to obtain a polymer solution containing polyvinylpyrrolidone having a thiol group at the terminal [hereinafter referred to as polymer (7)]. The solid content in the polymer solution was 22.7% by mass, and the weight average molecular weight of the polymer (7) was 5300.

Production Example 8
Add 180 g of isopropanol into a 500 mL reaction vessel equipped with a thermometer, stirrer, dropping funnel, nitrogen gas introduction tube and reflux condenser, and at a flow rate of 20 mL / min while refluxing the contents of the reaction vessel. The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel for 30 minutes.

Next, while maintaining the contents of the reaction vessel in a reflux state, a mixed solution of 100 g of N-vinylpyrrolidone, 0.4 g of 4,4'-azobis (4-cyanopentanoic acid) and 20 g of isopropanol was reacted over 2 hours. It was continuously dropped into the container. While maintaining the reflux state of the contents of the reaction vessel, 0.1 g of 4,4'-azobis (4-cyanopentanoic acid) was added 3 hours and 4 hours after the start of dropping the mixed solution, respectively, to further bring the reflux state. By maintaining for 2 hours, a polymer solution containing polyvinylpyrrolidone having a carboxyl group at the terminal [hereinafter referred to as polymer (8)] was obtained. The solid content in the polymer solution was 33.0% by mass, and the weight average molecular weight of the polymer (8) was 5200.

Manufacturing example 9
54 g of methoxypolyethylene glycol (average number of moles of oxyethylene group added: 120), tosyl lolide, in a 500 mL reaction vessel equipped with a thermometer, agitator, dropping funnel, nitrogen gas introduction tube and reflux condenser. 288 g, 1.52 g of triethylamine and 300 g of dichloromethane were added, and the contents of the reaction vessel were heated to 65 ° C.

Next, the contents of the reaction vessel were maintained at 65 ° C. for 24 hours, the salt contained in the contents was removed by filtration, and the obtained filtrate was desolvated under reduced pressure to form a tosylated product. Got 60 g of the obtained tosylated product, 1.37 g of potassium thioacetate and 300 g of acetonitrile were mixed, and the obtained mixture was maintained at 65 ° C. for 24 hours, and then the salt contained in the mixture was removed by filtration to reduce the pressure. An acetylated product was obtained by desolving underneath.

56 g of the acetylated product obtained above and 100 g of methanol were mixed, 25 g of a 1N aqueous sodium hydroxide solution was added to the obtained mixture, and the obtained mixture was stirred at a temperature of 65 ° C. for 10 minutes and then added to the mixture. 180 g of dichloromethane was added and extracted, and the organic layer was recovered to obtain methoxypolyethylene glycol having a thiol group at the terminal [hereinafter referred to as polymer (9)]. The solid content in the polymer solution was 30.0% by mass, and the weight average molecular weight of the polymer (9) was 5400.

Production Example 10
(1) Preparation of Polymer A 2 g of the polymer (3) obtained above and 2 g of dimethylformamide were added to a 100 mL Schlenk tube, and the obtained mixture was heated to 80 ° C.

Next, 170 mg of tert-butylpyrocarbonate and 50 mg of N, N-dimethyl-4-aminopyridine as a catalyst were added to the Schlenk tube, and the mixture was stirred for 8 hours, and then the obtained mixture was purified and freeze-dried. , A polymer A in which the carboxyl group of the polymer (3) was protected was obtained.

(2) Preparation of Polymer B 2 g of the polymer (6) obtained above and 2 g of dimethylformamide were added to a 100 mL Schlenk tube, and the obtained mixture was heated to 80 ° C.

Next, 170 mg of tert-butylpyrocarbonate and 100 mg of triethylamine were added into the Schlenk tube, and the mixture was stirred for 8 hours, and then the obtained mixture was purified and lyophilized to protect the amino group of the polymer (6). The polymer B was obtained.

(3) Preparation of Block Copolymer of Polymethoxyethyl Acrylate and Polyhydroxyethyl methacrylate Add 2 g of the polymer A obtained above, 2 g of the polymer B obtained above, and 4 g of dimethylformamide in a 100 mL Schlenk tube. , The resulting mixture was cooled to −5 ° C.

A mixture of 80 mg of dicyclohexylcarbodiimide and 1 g of dimethylformamide was added into a Schlenk tube under stirring while maintaining the temperature of the mixture at −5 ° C., maintained at −5 ° C. for 1 hour under stirring, and then at room temperature for 1 hour. I kept the time. Precipitates were removed by filtering the mixture, and 5 mL of trifluoroacetic acid was added to the filtrate under stirring while cooling the obtained filtrate in an ice-water bath to obtain a mixed solution.

Next, the mixed solution obtained above is stirred at 20 ° C. for 1 hour and freeze-dried to obtain polymethoxyethyl acrylate and polyhydroxy having a carboxyl group at one end and an amino group at the other end. A block copolymer with ethyl methacrylate [hereinafter referred to as polymer (10)] was obtained. The weight average molecular weight of the obtained polymer (10) was 10200.

Production Example 11
2 g of the polymer A obtained above, 2 g of the polymer (4) obtained above and 4 g of dimethylformamide were added to a 100 mL Schlenk tube, and the obtained mixture was cooled to −5 ° C.

A mixture of 80 mg of dicyclohexylcarbodiimide and 1 g of dimethylformamide was added into a Schlenk tube under stirring while maintaining the temperature of the mixture obtained above at −5 ° C., maintained at −5 ° C. for 1 hour under stirring, and then maintained at −5 ° C. for 1 hour. It was maintained at room temperature for 1 hour. Precipitates were removed by filtering the mixture, and 5 mL of trifluoroacetic acid was added to the filtrate under stirring while cooling the obtained filtrate in an ice-water bath to obtain a mixed solution.

Next, the mixed solution obtained above is stirred at 20 ° C. for 1 hour and freeze-dried to obtain polymethoxyethyl acrylate and polyhydroxy having a carboxyl group at one end and a thiol group at the other end. A block copolymer with ethyl methacrylate [hereinafter referred to as polymer (11)] was obtained. The weight average molecular weight of the obtained polymer (11) was 10400.

Production Example 12
2 g of the polymer (3) obtained above and 2 g of dimethylformamide were added to a 100 mL Schlenk tube, and the obtained mixture was heated to 80 ° C.

Next, 170 mg of tert-butylpyrocarbonate and 100 mg of triethylamine were added to the Schlenk tube, and the obtained mixture was stirred for 8 hours, purified, and freeze-dried to obtain polymer C having an amino group protected. Obtained.

Next, 2 g of the polymer A obtained above, 2 g of the polymer C obtained above and 4 g of dimethylformamide were added to a Schlenk tube having a volume of 100 mL different from the above, and the obtained mixture was cooled to −5 ° C. ..

A mixture of 80 mg of dicyclohexylcarbodiimide and 1 g of dimethylformamide was added into a Schlenk tube under stirring while maintaining the temperature of the mixture obtained above at −5 ° C., and maintained at −5 ° C. for 1 hour under stirring. It was then maintained at room temperature for 1 hour. Precipitates were removed by filtering the mixture, and 5 mL of trifluoroacetic acid was added to the filtrate under stirring while cooling the obtained filtrate in an ice-water bath, and the obtained mixed solution was added at 20 ° C. for 1 hour. The polymer D was obtained by stirring and lyophilizing.

Next, 4 g of the polymer D and 4 g of dimethylformamide were added into the Schlenk tube, and the obtained mixture was heated to 80 ° C., and then 340 mg of tert-butylpyrocarbonate and 100 mg of N, N-dimethyl-4-aminopyridine were added. It was added into a Schlenk tube and stirred for 8 hours. Then, the contents in the Schlenk tube were purified and freeze-dried to obtain a polymer E in which the carboxyl group was protected.

Next, 4 g of the polymer E obtained above, 2 g of the polymer B obtained above and 6 g of dimethylformamide were added to a Schlenk tube having a volume of 100 mL different from the above, and the obtained mixture was cooled to −5 ° C. did.

A mixture of 120 mg of dicyclohexylcarbodiimide and 1 g of dimethylformamide was added into a Schlenk tube under stirring while maintaining the temperature of the mixture at −5 ° C., maintained at −5 ° C. for 1 hour under stirring, and then at room temperature for 1 hour. I kept the time. Precipitates were removed by filtering the mixture, and 10 mL of trifluoroacetic acid was added to the filtrate under stirring while cooling the obtained filtrate in an ice-water bath, followed by stirring at 20 ° C. for 1 hour, and freeze-dried. The polymer F was obtained.

Next, 6 g of the polymer F and 6 g of dimethylformamide obtained above were added to another 100 mL Schlenk tube, and the obtained mixture was heated to 80 ° C., and then tert-butylpyrocarbonate 540 mg. And 150 mg of N, N-dimethyl-4-aminopyridine were added into the Schlenk tube and stirred for 8 hours. Then, the contents in the Schlenk tube were purified and freeze-dried to obtain a polymer G having a protected carboxyl group.

Next, 6 g of the polymer G obtained above, 2 g of the polymer (4) obtained above and 8 g of dimethylformamide were added to a 100 mL Schlenk tube different from the above, and the obtained mixture was added to −5 ° C. A mixture of 160 mg of dicyclohexylcarbodiimide and 1 g of dimethylformamide was added to the mixture under stirring while maintaining a temperature of -5 ° C, and the mixture was stirred for 1 hour while maintaining a temperature of -5 ° C. Then it was stirred at room temperature for 1 hour. Precipitates are removed by filtering the mixture, and 15 mL of trifluoroacetic acid is added to the filtrate under stirring while cooling the obtained filtrate in an ice-water bath, followed by stirring at 20 ° C. for 1 hour, and freeze-drying. A block copolymer of polymethoxyethyl acrylate having a carboxyl group at one end and a thiol group at the other end and polyhydroxyethyl methacrylate [hereinafter referred to as polymer (12)] was obtained. .. The weight average molecular weight of the obtained polymer (12) was 21000.

Production Example 13
180 g of isopropanol was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the isopropanol in the reaction vessel was refluxed at a flow rate of 20 mL / min. The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel for 30 minutes. Then, a mixed solution of 100 g of 2-hydroxyethyl methacrylate, 0.4 g of 4,4'-azobis (4-cyanopentanoic acid) and 20 g of isopropanol was continuously added dropwise into the reaction vessel over 2 hours.

Next, while maintaining the reflux state, 0.1 g of 4,4'-azobis (4-cyanopentanoic acid) was added into the reaction vessel 3 hours and 4 hours after the start of dropping the mixed solution, respectively, to bring the reflux state into a reflux state. By maintaining for 2 hours, a polymer solution containing a polyhydroxyethyl acrylate having a carboxyl group at the terminal [hereinafter referred to as a polymer (13)] was obtained. The solid content in the polymer solution was 33.0% by mass, and the weight average molecular weight of the polymer (13) was 5700.

Production example 14
180 g of isopropanol was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the isopropanol in the reaction vessel was refluxed at a flow rate of 20 mL / min. The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel for 30 minutes. Then, a mixed solution of 100 g of 2-hydroxyethyl acrylate, 0.4 g of 4,4'-azobis (4-cyanopentanoic acid) and 20 g of isopropanol was continuously added dropwise into the reaction vessel over 2 hours.

Next, while maintaining the reflux state, 0.1 g of 4,4'-azobis (4-cyanopentanoic acid) was added into the reaction vessel 3 hours and 4 hours after the start of dropping the mixed solution, respectively, to bring the reflux state into a reflux state. By maintaining for 2 hours, a polymer solution containing a polyhydroxyethyl acrylate having a carboxyl group at the terminal [hereinafter referred to as a polymer (14)] was obtained. The solid content in the polymer solution was 32.5% by mass, and the weight average molecular weight of the polymer (14) was 5200.

Production example 15
180 g of isopropanol was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the isopropanol in the reaction vessel was refluxed at a flow rate of 20 mL / min. The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel for 30 minutes. Then, a mixed solution of 100 g of N-vinylpyrrolidone, 0.4 g of 4,4'-azobis (4-cyanopentanoic acid) and 20 g of isopropanol was continuously added dropwise into the reaction vessel over 2 hours.

Next, while maintaining the reflux state, 0.1 g of 4,4'-azobis (4-cyanopentanoic acid) was added into the reaction vessel 3 hours and 4 hours after the start of dropping the mixed solution, respectively, to bring the reflux state into a reflux state. By maintaining for 2 hours, a polymer solution containing poly N-vinylpyrrolidone having a carboxyl group at the terminal [hereinafter referred to as polymer (15)] was obtained. The solid content in the polymer solution was 32.8% by mass, and the weight average molecular weight of the polymer (15) was 5400.

Production example 16
180 g of isopropanol was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the isopropanol in the reaction vessel was refluxed at a flow rate of 20 mL / min. The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel for 30 minutes. Then, 100 g of methoxypolyethylene glycol methacrylate (average number of moles of ethylene oxide added: 9), 0.4 g of 4,4'-azobis (4-cyanopentanoic acid) and 20 g of isopropanol were continuously placed in the reaction vessel over 2 hours. Dropped.

Next, while maintaining the reflux state, 0.1 g of 4,4'-azobis (4-cyanopentanoic acid) was added into the reaction vessel 3 hours and 4 hours after the start of dropping the mixed solution, respectively, to bring the reflux state into a reflux state. By maintaining for 2 hours, a polymer solution containing polymethoxypolyethylene glycol methacrylate having a carboxyl group at the terminal [hereinafter referred to as polymer (16)] was obtained. The solid content in the polymer solution was 32.7% by mass, and the weight average molecular weight of the polymer (16) was 5000.

Production example 17
180 g of isopropanol was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the isopropanol in the reaction vessel was refluxed at a flow rate of 20 mL / min. The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel for 30 minutes. Then, a mixed solution of 100 g of 2-methacryloyloxyethyl phosphorylcholine, 0.4 g of 4,4'-azobis (4-cyanopentanoic acid) and 20 g of isopropanol was continuously added dropwise into the reaction vessel over 2 hours.

Next, while maintaining the reflux state, 0.1 g of 4,4'-azobis (4-cyanopentanoic acid) was added into the reaction vessel 3 hours and 4 hours after the start of dropping the mixed solution, respectively, to bring the reflux state into a reflux state. By maintaining for 2 hours, a polymer solution containing poly2-methacryloyloxyethyl phosphorylcholine [hereinafter referred to as polymer (17)] having a carboxyl group at the terminal was obtained. The solid content in the polymer solution was 33.0% by mass, and the weight average molecular weight of the polymer (17) was 5000.

Production Example 18
180 g of isopropanol was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the isopropanol in the reaction vessel was refluxed at a flow rate of 20 mL / min. The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel for 30 minutes. Then, a mixed solution of 100 g of tetrahydrofurfuryl acrylate, 0.4 g of 4,4'-azobis (4-cyanopentanoic acid) and 20 g of isopropanol was continuously added dropwise into the reaction vessel over 2 hours.

Next, while maintaining the reflux state, 0.1 g of 4,4'-azobis (4-cyanopentanoic acid) was added into the reaction vessel 3 hours and 4 hours after the start of dropping the mixed solution, respectively, to bring the reflux state into a reflux state. By maintaining for 2 hours, a polymer solution containing a polytetrahydrofurfuryl acrylate having a carboxyl group at the terminal [hereinafter referred to as a polymer (18)] was obtained. The solid content in the polymer solution was 33.1% by mass, and the weight average molecular weight of the polymer (18) was 5200.

Production example 19
180 g of isopropanol was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the temperature of the isopropanol in the reaction vessel was raised to 70 ° C. at 20 mL / The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel at a flow rate of min for 30 minutes. Then, while maintaining the internal temperature at 70 ° C., a mixed solution of 100 g of N-vinylpyrrolidone, 0.4 g of 2,2'-azobis [N- (2-carboxyethyl) -2-methylpropion amidine] and 20 g of isopropanol was added. It was continuously added dropwise into the reaction vessel over time.

Next, while maintaining the internal temperature at 70 ° C., 0.1 g of azoisobutyronitrile was added 3 hours and 4 hours after the start of dropping of the mixed solution, respectively, and the reflux state was maintained for 2 hours to obtain carboxyl at the terminal. A polymer solution containing polyvinylpyrrolidone having a group [hereinafter referred to as polymer (19)] was obtained. The solid content in the polymer solution was 32.4% by mass, and the weight average molecular weight of the polymer (19) was 5600.

Production example 20
Add 180 g of ethanol into a 500 mL reaction vessel equipped with a thermometer, stirrer, dropping funnel, nitrogen gas introduction tube and reflux condenser, and raise the temperature of ethanol in the reaction vessel to 70 ° C. at 20 mL / The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel at a flow rate of min for 30 minutes. Then, while maintaining the internal temperature at 70 ° C., a mixed solution of 100 g of N-vinylpyrrolidone, 0.4 g of 2,2'-azobis (propane-2-carboamidine) dihydrochloride and 20 g of ethanol was placed in a reaction vessel over 2 hours. It was continuously dropped inside.

Next, while maintaining the internal temperature at 70 ° C., 0.1 g of azoisobutyronitrile was added 3 hours and 4 hours after the start of dropping of the mixed solution, respectively, and the reflux state was maintained for 2 hours to achieve amino at the terminal. A polymer solution containing polyvinylpyrrolidone having a group [hereinafter referred to as polymer (20)] was obtained. The solid content in the polymer solution was 32.0% by mass, and the weight average molecular weight of the polymer (20) was 4800.

Production example 21
180 g of ethanol was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the temperature of the ethanol in the reaction vessel was raised to 70 ° C. at 20 mL / The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel at a flow rate of min for 30 minutes. Then, while maintaining the internal temperature at 70 ° C., 100 g of N-vinylpyrrolidone, 0.4 g of 2,2'-bis (2-imidazolin-2-yl) [2,2'-azobispropane] dihydrochloride and ethanol. 20 g of the mixed solution was continuously added dropwise into the reaction vessel over 2 hours.

Next, while maintaining the internal temperature at 70 ° C., 0.1 g of azoisobutyronitrile was added 3 hours and 4 hours after the start of dropping of the mixed solution, respectively, and the reflux state was maintained for 2 hours to achieve amino at the terminal. A polymer solution containing polyvinylpyrrolidone having a group [hereinafter referred to as polymer (21)] was obtained. The solid content in the polymer solution was 32.7% by mass, and the weight average molecular weight of the polymer (21) was 4700.

Production example 22
180 g of isopropanol was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the isopropanol in the reaction vessel was refluxed at a flow rate of 20 mL / min. The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel for 30 minutes. Then, a mixed solution of 100 g of N-vinylpyrrolidone, 0.8 g of 2,2'-azobismethyl-N- [1,1-bis (hydroxymethyl) -2-hydroxyethyl] propionamide] and 20 g of isopropanol was added over 2 hours. It was continuously added dropwise into the reaction vessel.

Next, while maintaining the reflux state, 0.1 g of azoisobutyronitrile was added 3 hours and 4 hours after the start of dropping of the mixed solution, respectively, and the reflux state was maintained for 2 hours to add a hydroxy group to the terminal. A polymer solution containing polyvinylpyrrolidone [hereinafter referred to as polymer (22)] was obtained. The solid content in the polymer solution was 33.0% by mass, and the weight average molecular weight of the polymer (22) was 4900.

Production Example 23
180 g of isopropanol was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the isopropanol in the reaction vessel was refluxed at a flow rate of 20 mL / min. The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel for 30 minutes. Then, a mixed solution of 100 g of N-vinylpyrrolidone, 0.4 g of 2,2'-azobis [N- (2-hydroxyethyl) -2-methylpropanamide] and 20 g of isopropanol was continuously added dropwise into the reaction vessel over 2 hours. did.

Next, while maintaining the reflux state, 0.1 g of azoisobutyronitrile was added 3 hours and 4 hours after the start of dropping of the mixed solution, respectively, and the reflux state was maintained for 2 hours to add a hydroxy group to the terminal. A polymer solution containing polyvinylpyrrolidone [hereinafter referred to as polymer (23)] was obtained. The solid content in the polymer solution was 32.4% by mass, and the weight average molecular weight of the polymer (23) was 5000.

Production example 24
180 g of isopropanol was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the temperature of the isopropanol in the reaction vessel was raised to 70 ° C. at 20 mL / The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel at a flow rate of min for 30 minutes. Then, while maintaining the internal temperature at 70 ° C., 100 g of N-vinylpyrrolidone, 2,2'-azobis [2- [1- (2-hydroxyethyl) -2-imidazolin-2-yl] propane] dihydrochloride 0 A mixture of 0.8 g and 20 g of isopropanol was continuously added dropwise into the reaction vessel over 2 hours.

Next, while maintaining the internal temperature at 70 ° C., 0.1 g of azoisobutyronitrile was added 3 hours and 4 hours after the start of dropping of the mixed solution, respectively, and the reflux state was maintained for 2 hours to obtain hydroxy at the terminal. A polymer solution containing polyvinylpyrrolidone having a group [hereinafter referred to as polymer (24)] was obtained. The solid content in the polymer solution was 33.3% by mass, and the weight average molecular weight of the polymer (24) was 5200.

Production example 25
180 g of isopropanol was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the isopropanol in the reaction vessel was refluxed at a flow rate of 20 mL / min. The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel for 30 minutes. Then, a mixed solution of 100 g of N-vinylpyrrolidone, 0.4 g of 2,2'-azobis (2-methyl-2-propenylpropanamide) and 20 g of isopropanol was continuously added dropwise into the reaction vessel over 2 hours.

Next, while maintaining the reflux state, 0.1 g of azoisobutyronitrile was added 3 hours and 4 hours after the start of dropping of the mixed solution, respectively, and the reflux state was maintained for 2 hours to add an allyl group to the terminal. A polymer solution containing polyvinylpyrrolidone [hereinafter referred to as polymer (25)] was obtained. The solid content in the polymer solution was 32.8% by mass, and the weight average molecular weight of the polymer (25) was 5100.

Production example 26
180 g of toluene was added to a 500 mL reaction vessel equipped with a thermometer, a stirrer, a dropping funnel, a nitrogen gas introduction tube and a reflux condenser, and the temperature of the toluene in the reaction vessel was raised to 90 ° C. to 20 mL / min. The internal space of the reaction vessel was replaced with nitrogen gas by introducing nitrogen gas into the reaction vessel at the above flow rate for 30 minutes.

Next, a mixed solution of 100 g of methyl methacrylate, 0.05 g of azobisisobutyronitrile and 20 g of toluene was continuously added dropwise to a reaction vessel kept at 90 ° C. over 2 hours. While maintaining the temperature of the contents in the reaction vessel at 90 ° C., 0.01 g of azobisisobutyronitrile was added into the reaction vessel 3 hours and 4 hours after the start of dropping of the mixed solution, respectively, and then the reaction vessel was charged. By maintaining the temperature of the contents at 90 ° C. for 2 hours, a polymer solution containing polymethylmethacrylate (hereinafter referred to as polymer (26)) was obtained. The solid content in the polymer solution was 33.1% by mass, and the weight average molecular weight of the polymer (26) was 71000.

Example 1
After the polymer (1) obtained above is freeze-dried for 8 hours, 0.05 mmol of the polymer (1), 0.5 mmol of hydrogenated soy lecithin, 0.37 mmol of cholesterol and 3,5-dipentadecyloxy A lipid mixture prepared by dissolving 0.08 mmol of benzamidine hydrochloride in 30 mL of tert-butyl alcohol is frozen in an ice bath and freeze-dried for about 8 hours to prepare a lipid mixture (a lipid mixture that is a raw material for the lipid membrane of liposomes). 1) was obtained.

The inner aqueous layer of liposomes (2) by dissolving 0.2 mmol of the fluorescent dye carboxyfluoroscein in 10 mL of 0.01 mol / L phosphate buffered saline [manufactured by Wako Pure Chemical Industries, Ltd.] (hereinafter referred to as PBS). ) Was obtained.

Next, the inner aqueous layer (2) is added to the mixture (1) obtained above, the mixed solution is heated at 65 ° C. for 10 minutes, and then the mixed solution is subjected to ultrasonic treatment for 5 minutes. , A liposome dispersion was obtained. The liposome dispersion obtained above was pressure-fed to a polycarbonate film having a pore size of 0.2 μm, and further pressure-fed to a polycarbonate film having a pore size of 0.1 μm to perform granulation. The average particle size of the liposomes obtained above was measured using a concentrated particle size analyzer [manufactured by Otsuka Electronics Co., Ltd., product number: FPAR-1000]. The results are shown in Table 1.

Next, using the liposomes or liposome dispersions obtained above, the sustained release property, particle shape retention, hemolytic resistance and in vitro release property of the inclusions were examined based on the following methods. The results are shown in Table 1.

(1) Sustained release of drug 1 mL of liposome dispersion and 200 μL of rabbit defibered blood [manufactured by Koji Bio Co., Ltd.] and 1 mL of PBS are placed in a constant temperature bath at 37 ° C., incubated for 30 minutes or 24 hours, and then corona. Using SH-9000 manufactured by Denki Co., Ltd., the fluorescence intensity was measured at an excitation wavelength of 494 nm and a fluorescence wavelength of 521 nm, and the release rate of the drug was expressed by the formula:
[Drug release rate (%)] = [(F _2- F ₁ ) / (F _3- F ₁ )] × 100
[In the formula, F ₁ is the fluorescence intensity before incubation, F ₂ is the fluorescence intensity after incubation at 37 ° C., and F ₃ is 2% polyoxyethylene (10) octylphenyl ether instead of PBS added to the mixed solution. [Manufactured by Wako Pure Chemical Industries, Ltd., trade name: Triton-X] Shows fluorescence intensity (100% leakage) after adding and mixing PBS solution]
The sustained release of the drug was evaluated based on the following evaluation criteria.

[Evaluation criteria for sustained release of inclusions]
A. When the incubation time is 30 minutes ⊚: Drug release rate is less than 10% ○: Drug release rate is 10% or more and less than 30% Δ: Drug release rate is 30% or more and less than 90% ×: Drug release rate Is 90% or more

B. When the incubation time is 24 hours ⊚: Drug release rate is less than 60% ○: Drug release rate 30% or more and less than 60% Δ: Drug release rate is 10% or more and less than 30% ×: Drug release rate Is 10% or more

(2) Retention of particle shape A parallel plate with a diameter of 7.9 mm of a viscoelasticity measuring device [manufactured by Leometric Scientific FE Co., Ltd.] was set in a sample holder with a void of 10 μm. After adding 1 mL of the liposome dispersion and applying viscoelasticity at a frequency of 1 Hz for 5 minutes, the liposome dispersion was collected and the average particle size was measured, and the maintenance rate of the particle size was calculated.
[Maintenance rate of particle size (%)] = [Average particle size after test / Average particle size before test] x 100
The particle shape retention was evaluated based on the following evaluation criteria.

[Evaluation criteria for particle shape retention]
◯: Particle diameter maintenance rate is 70% or more and 100% or less Δ: Particle diameter maintenance rate is 20% or more and less than 70% ×: Particle diameter maintenance rate is less than 20%

(3) Hemolytic resistance Liposomes were dissolved in PBS to prepare a 1% by mass PBS solution.

48 mL of PBS solution was added to 2 mL of rabbit defibered blood [manufactured by Kojin Bio Co., Ltd.] and mixed by inversion to prepare a mixed solution, which was then centrifuged at 2000 rpm for 10 minutes at 4 ° C. A blood solution was obtained by adding 48 mL of PBS to the mixed solution from which the supernatant had been removed and mixing by inversion. To 2 mL of the obtained blood solution, 2 mL of the liposome solution obtained above was added, mixed by inversion, incubated at 37 ° C. for 1 hour, and further centrifuged at 2000 rpm for 10 minutes at 4 ° C., and then the wavelength was 545 nm. The absorbance of the supernatant was measured with a spectrophotometer [manufactured by Shimadzu Corporation, product number: UV-3600].

2 mL of 2% polyoxyethylene (10) octylphenyl ether [manufactured by Wako Pure Chemical Industries, Ltd., trade name: Triton-X] PBS solution was added to 2 mL of the blood solution obtained above, and the solution was inverted and mixed. The absorbance at a wavelength of 545 nm was measured in the same manner as described above. The absorbance at this time was 100% hemolytic.

Next, formulate the degree of hemolysis:
[Hemolysis degree (%)] = [Absorvity of incubation supernatant / Absorbance when using Triton-X] x 100
The hemolytic resistance was evaluated based on the following evaluation criteria.

[Evaluation criteria for hemolytic resistance]
◯: Hemolysis rate is less than 30% Δ: Hemolysis rate is 30% or more and less than 70% ×: Hemolysis rate is 70% or more

(4) In vitro release property A mixture was obtained by mixing 0.1 g of the polymer freeze-dried for 8 hours and 0.5 g of dimethylformamide.

Next, a mixed solution was prepared by adding 1 g of rabbit defibered blood [manufactured by Kojin Bio Co., Ltd.] and 3 g of PBS to the mixture obtained above. The obtained mixed solution was inverted and mixed, and then incubated at 37 ° C. for 24 hours.

Next, the mixed solution incubated above was centrifuged at 2000 rpm for 10 minutes at 4 ° C., and the supernatant was diluted 10-fold with dimethylformamide, and gel permeation chromatography was performed in the same manner as described above. The weight average molecular weight of the polymer was measured, and the in vitro release property was evaluated using the weight average molecular weight of the polymer as an index. The evaluation criteria are shown below. The lower the weight average molecular weight of the polymer, the easier it is for the polymer to be released from the body.

[Evaluation criteria for extracorporeal release]
⊚: Weight average molecular weight of polymer is 8000 or less ◯: Weight average molecular weight of polymer is more than 8000, 30,000 or less Δ: Weight average molecular weight of polymer is more than 30,000, 60,000 or less ×: Weight average molecular weight of polymer is more than 8000 Over 60,000

Examples 2 to 25 and Comparative Example 1
Liposomes and liposome dispersions were prepared in the same manner as in Example 1 except that the polymers shown in Table 1 were used instead of the polymer 1. The average particle size of the liposomes obtained above was measured using a concentrated particle size analyzer [manufactured by Otsuka Electronics Co., Ltd., product number: FPAR-1000]. The results are shown in Table 1.

Next, using the liposomes or liposome dispersions obtained above, the sustained release property, particle shape retention, hemolytic resistance and in vitro release property of the inclusions were examined based on the following methods. The results are shown in Table 1.

Example 26
After freeze-drying the polymer (12) obtained above for 8 hours, 0.6 mmol of the polymer (12) and 0.08 mmol of 3,5-dipentadecyloxybenzamidine hydrochloride were added to 30 mL of tert-butyl alcohol. The mixed solution prepared by dissolving in was frozen in an ice bath and freeze-dried for about 8 hours to obtain a lipid mixture (1) as a raw material for polymer micelles.

Inclusion-containing solution (2) by dissolving 0.2 mmol of the fluorescent dye carboxyfluoroscein in 10 mL of 0.01 mol / L phosphate buffered saline [manufactured by Wako Pure Chemical Industries, Ltd.] (hereinafter referred to as PBS). Got

Next, the solution (2) is added to the mixture (1) obtained above, the mixed solution is heated at 65 ° C. for 10 minutes, and then the mixed solution is subjected to ultrasonic treatment for 5 minutes. A molecular micelle solution was obtained. Granulation was performed by pressure-feeding the dispersion liquid obtained above to a polycarbonate film having a pore size of 0.05 μm. The average particle size of the polymer micelles obtained above was measured using a concentrated particle size analyzer [manufactured by Otsuka Electronics Co., Ltd., product number: FPAR-1000]. The results are shown in Table 1.

Next, using the polymer micelle dispersion obtained above, the sustained release property, particle shape retention, hemolytic resistance and extracorporeal release property of the inclusions were examined based on the following methods. The results are shown in Table 1.

From the results shown in Table 1, the biocompatible medical materials obtained in each example are excellent in sustained release of inclusions, retention of the shape of formed particles, hemolytic resistance and extracorporeal release. You can see that. Therefore, the biocompatible medical material of the present invention is, for example, a medical base material used for a carrier, a medical device, an instrument, etc. used when holding a drug and introducing it into the body orally or via blood. It is expected to be used as a coating agent for coating agents.

Claims (2)

A biocompatible medical material comprising a biocompatible polymer obtained by polymerizing a biocompatible monomer, having a weight average molecular weight of 1000 to 90000, and having a functional group at at least one molecular terminal. wherein the biocompatible polymer as alkoxyalkyl (meth) comprising acrylate is polymerized polymer is used, the functional groups at the molecular ends of the polymer has the formula: -COOM (M represents a hydrogen atom or an alkali metal atom) a group represented by a hydroxyl group, an allyl group, an epoxy group, an aldehyde group, -NH ₂ group, CONH- group or -SH Motodea Ru biocompatible medical material. A biocompatible polymer obtained by polymerizing the biocompatible polymer according to claim 1 and a hydroxyl group-containing (meth) acrylate, having a weight average molecular weight of 1000 to 90000 and having a functional group at at least one molecular terminal. A biocompatible medical material made by combining with.