WO2017191827A1

WO2017191827A1 - Internally degradable polyrotaxane and synthesis method therefor

Info

Publication number: WO2017191827A1
Application number: PCT/JP2017/017133
Authority: WO
Inventors: 由井　伸彦; 篤志田村; 慶紀有坂; 麻由伏見
Original assignee: 国立大学法人東京医科歯科大学
Priority date: 2016-05-02
Filing date: 2017-05-01
Publication date: 2017-11-09
Also published as: JPWO2017191827A1

Abstract

The present invention provides a degradable polyrotaxane compound that is easily produced and has large changes in physical properties during degradation. More specifically, said polyrotaxane compound is provided as a result of synthesizing a polyrotaxane compound including at least two linear polymer moieties wherein linear polymers are linked via at least one degradable section. This kind of polyrotaxane compound has a degradable section inside a linear polymer main chain and, as a result, is capable of having a greater change in molecular weight during degradation, compared to conventional degradable polyrotaxane compounds having degradable bonds at both terminals thereof, and is capable of dramatically changing basic physical properties of the polymer such as viscosity, solubility, and glass transition point.

Description

Internally decomposed polyrotaxane and method for synthesizing the same

Cross-reference of related applications

This application is an application that enjoys the benefit of the priority of Japanese Patent Application No. 2016-92550 (filing date: May 2, 2016), which is incorporated herein by reference in its entirety.

The present invention relates to a degradable polyrotaxane compound having at least one degradable bond inside a linear polymer, and a synthesis method thereof.

Polyrotaxane is a supramolecule having a skeleton in which a linear polymer penetrates a plurality of cyclic molecular cavities. For example, when polyethylene glycol (PEG) (linear polymer) with high biocompatibility and cyclodextrin (CD) (cyclic molecule) used in medicine and food are mixed in an aqueous solution, PEG spontaneously becomes CD. It is known that the inclusion complex penetrating through the cavity can be recovered as a precipitate. Moreover, a polyrotaxane can be prepared by modifying a bulky functional group (blocking group) at the end of the linear polymer of the inclusion complex.

In International Publication Pamphlet WO2015 / 025815 (Patent Document 1), a polyrotaxane in which a functional group containing a biodegradable bond is arranged between both ends of a linear polymer and a blocking group is synthesized. It has been shown that the degradation of the entire polyrotaxane is induced with dissociation. Conventional degradable polymers have the property that only a part modified with a degradable group is sequentially decomposed to lower the molecular weight over time (Non-patent Documents 1 and 2), but degradable polyrotaxanes are produced by dissociation of blocking groups. It has a decomposition characteristic accompanied by a large molecular weight change in which a cyclic molecule is liberated from a linear polymer. By selecting such a degradable polyrotaxane degradable functional group depending on the purpose, it is possible to utilize this degradable polyrotaxane in various environments such as in vivo. Further, CD in which glucose is cyclically bonded with α-1,4 bonds has a large number of hydroxyl groups, so that various functional groups can be modified to polyrotaxane, and the polyrotaxane can be easily functionalized.

International Publication No. 2015/025815

An object of the present invention is to provide a degradable polyrotaxane compound having at least one decomposable bond inside a linear polymer. Another object of the present invention is to provide a method for producing a degradable polyrotaxane compound having at least one degradable bond inside a linear polymer. All of the degradable polyrotaxane compounds reported so far have degradable bonds at both ends of the linear polymer chain, and two degradable bonds must be arranged for one molecule of polyrotaxane. Therefore, the synthesis method required 6 to 7 steps or more. Another object of the present invention is to provide a composition for various uses, which contains a degradable polyrotaxane compound having at least one degradable bond inside a linear polymer.

The present inventors have synthesized a linear polymer in which a functional group containing a bond that can be decomposed in an in vivo environment is arranged in the main chain away from the terminal site, and this linear polymer is a large number of cyclic molecules. We succeeded in obtaining polyrotaxane, a supramolecule penetrating through the cavity of (for example, cyclodextrin) with high purity and high yield. This polyrotaxane has a new dissociation mechanism in which all cyclic molecules are released from the linear polymer as the degradable part (degradable bond) in the linear polymer is decomposed by an external stimulus. Since the type of degradable bond can be selected to be decomposed as needed for treatment or diagnosis at any site / environment such as tissue or intracellular location, various stimuli in specific organelles (specific enzymes, etc.) It is possible to precisely prepare an intracellular degradable polyrotaxane that dissociates in response to. In addition, a wide variety of biodegradable three-dimensional cross-linked bodies having a polyrotaxane skeleton can be prepared based on modifications such as polymerizable groups to cyclodextrin (CD), so that they can be cross-linked when applied to tissue regeneration or dental treatment. Physical properties of the body (mechanical strength, tissue adhesion, cell differentiation / proliferation, etc.) can be freely reduced according to the healing process and therapeutic purpose of the application site.

Aspects of the present invention relate to the following matters.
[1] comprising at least two linear polymer portions each including a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymers are linked via at least one degradable portion. A polyrotaxane compound.
[2] The linear polymer has the following formula:
^{^{^{^{X 1 -Y 0 (-Z i -Y}}}} i) i -X 2
Wherein X ¹ and X ² are the same or different end groups, Y ⁰ and Y ⁱ are the same or different linear polymer moieties, and Z ⁱ is a degradable moiety , (-Z ⁱ -Y ⁱ ) _i indicates that there are i repeating units composed of a degradable portion Y and a linear polymer portion Z, and each Z ⁱ and Y ⁱ in the linear polymer is the same However, the polyrotaxane compound according to [1], wherein i is an integer of 1 to 500.
[3] The linear polymer has the following formula:
X ¹ -Y ⁰ -Z ¹ -Y ¹ -X ²
Has a structure shown in, wherein, X ¹ and X ² are the same or different terminal group, Y ⁰ and Y ¹ are identical or different linear polymeric moiety, Z ¹ is a degradable moiety The polyrotaxane compound according to [1].
[4] The polyrotaxane according to any one of [1] to [3], wherein the ratio of the lengths of two linear polymer moieties linked to the degradable moiety is included in 1: 1 to 1: 4. Compound.
[5] The polyrotaxane compound according to any one of [1] to [4], wherein the degradable moiety is acid decomposable, enzymatic degradable, thermally degradable, or photodegradable.
[6] The decomposable moiety includes at least one decomposable group, and the decomposable group includes a p-methoxyphenacyl group, a 2-nitrobenzyl group, a 2-nitrobenzyloxycarbonyl group, and 2-nitrophenylethylene. Glycol group, benzyloxycarbonyl group, 3,5-dimethoxybenzyloxycarbonyl group, α, α-dimethyl-3,5-dimethoxybenzyloxycarbonyl group, 3-nitrophenyl group, 3-nitrophenoxy group, 3,5- Dinitrophenoxy group, 3-nitrophenoxycarbonyl group, phenacyl group, 4-methoxyphenacyl group, α-methylphenacyl group, 3,5-dimethoxybenzoinyl group, 2,4-dinitrobenzenesulfenyl group, (coumarin -4-yl) methyl group, 7-nitroindolinyl group, arylazophosphate, [1] to [5] selected from the group consisting of a tellurium bond, a Schiff base bond, a carbamate bond, a peptide bond, an ether bond, an acetal bond, a hemiacetal bond, a disulfide bond, an organic peroxide, and an acylhydrazine bond. The polyrotaxane compound according to any one of the above.
[7] The polyrotaxane compound according to any one of [1] to [6], wherein the cyclic molecule is selected from the group consisting of α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin.
[8] The polyrotaxane compound according to any one of [1] to [7], wherein the cyclic molecule has one or more substituents.
[9] The substituent is 2-hydroxyethoxyethyl (HEE), hydroxyethyl, hydroxypropyl, hydroxybutyl, hydroxyethoxyethyl, N, N-dimethylaminoethyl, carboxyl, methyl, sulfo Group, primary amino group, polyethylene glycol, collagen, transferrin, RGD peptide, oligoarginine, (meth) acryloyl group, (meth) acryloyloxy group, (meth) acryloylamino group, (meth) acryloylthio group, vinyl group, aryl group The polyrotaxane compound according to [8], which is selected from the group consisting of a group, a styryl group, and a (meth) acrylamide group.
[10] The linear polymer portion is selected from the group consisting of polyethylene glycol, polypropylene glycol, a copolymer of polyethylene glycol and polypropylene glycol, polyethyleneimine, polyamino acid, and polymethyl vinyl ether, [1] to [9] ] The polyrotaxane compound as described in any one of.
[11] The terminal groups are 2,4-dinitrophenyl group, 3,5-dinitrophenyl group, cyclodextrin, adamantane group, O-triphenylmethyl (O-Trt) group, S-triphenylmethyl (S-Trt) ) Group, N-triphenylmethyl (N-Trt) group, N-tritylglycine, benzyloxycarbonyl (Z) group, 9-fluorenylmethyloxycarbonyl (Fmoc) group, benzyl ester (OBz) group, third group Butylcarbonyl (Boc) group, amino acid tertiary butyl ester (OBu group), trityl group, fluorescein, pyrene, substituted benzene, optionally substituted polynuclear aromatic, MPC (2-methacryloyloxyethyl phosphorylcholine), BMA (n -Butyl methacrylate), a combination of MPC and BMA, and steroids Is selected from the al group consisting, [1] ~ polyrotaxane compound according to any one claim of [10].
[12] comprising at least two linear polymer portions each including a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymers are linked via at least one degradable portion. An adhesive composition containing a polyrotaxane compound.
[13] comprising at least two linear polymer portions each including a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymers are linked via at least one degradable portion. A dental material composition containing a polyrotaxane compound.
[14] comprising at least two linear polymer portions each including a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymers are linked via at least one degradable portion. A surface coating agent containing a polyrotaxane compound.
[15] comprising at least two linear polymer portions each including a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymers are linked via at least one degradable portion. An anti-adhesion agent comprising a polyrotaxane compound.
[16] At least two linear polymer portions each including a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymers are linked via at least one degradable portion. A body implant containing a polyrotaxane compound.
[17] At least two linear polymer portions each including a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymers are linked via at least one degradable portion. A tissue regeneration device containing a polyrotaxane compound.
[18] comprising at least two linear polymer portions each including a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymers are linked via at least one degradable portion. A pharmaceutical composition for use in treating or preventing a disease, comprising a polyrotaxane compound.
[19] For a subject, preferably for a human, comprising a plurality of cyclic molecules and one linear polymer having a terminal group, the linear polymer via at least one degradable moiety A method for the treatment or prophylaxis of a disease comprising the step of administering a pharmaceutical composition comprising a polyrotaxane compound comprising at least two linear polymer moieties linked.
[20] In the manufacture of a medicament for the treatment or prevention of a disease, it comprises a plurality of cyclic molecules and one linear polymer having a terminal group, and the linear polymer passes through at least one degradable moiety. Use of a polyrotaxane compound comprising at least two linear polymer moieties linked together.
[21] The pharmaceutical composition according to [18], wherein the disease is a disease caused by an abnormal cell metabolic function, a disease caused by intracellular cholesterol accumulation, or a disease caused by a dysfunction of autophagy, [19] Or the use according to [20].
[22] The pharmaceutical composition according to [18], the method according to [19], or the use according to [20], wherein the disease is selected from the group consisting of lysosomal disease, neurodegenerative disease, and cancer.
[23] The diseases are Gaucher disease (Gaucher disease), Niemann-Pick disease type A (Niemann-Pick disease type A), Niemann-Pick disease type B (Niemann-Pick disease type B), Niemann-Pick disease type C (Niemann-Pick disease type C), GM1 gangliosidosis, GM2 gangliosidosis, Krabbe disease (Krabbe disease), metachromatic leukodystrophy, multiple sulfatase deficiency, Farber disease (Farber disease) Disease), mucopolysaccharidosis type I, mucopolysaccharidosis type II, mucopolysaccharidosis type III, mucopolysaccharidosis type IV, mucopolysaccharidosis type VI, mucopolysaccharidosis type VII, mucopolysaccharidosis type IX, sialidosis, galactosialidosis , I-cell disease / mucolipidosis type III, α-mannosidosis , Β-mannosidosis, fucosidosis, aspartylglucosamineuria, Schindler / Kanzaki disease (Schindler / Kanzaki disease), Wolman disease (Wolman disease), Danone disease (Danon disease), free sialic acid storage disease, ceroid lipofus The pharmaceutical composition according to [18], the method according to [19], or the method according to [20], selected from the group consisting of sclerosis, Fabry disease, Alzheimer's disease, Parkinson's disease, Huntington's disease, and apoptosis-resistant cancer use.
[24] At least two linear polymer portions each including a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymers are linked via at least one degradable portion. A method for producing a polyrotaxane compound comprising: a) adding a reactive group to both ends of a linear polymer portion; b) decomposing a linear polymer portion having a reactive group added to both ends; A linear polymer comprising at least two linear polymer moieties linked via at least one degradable moiety, c) reacting the linear polymer with a cyclic molecule A process for obtaining a pseudopolyrotaxane, and d) a step of adding end groups to both ends of the pseudopolyrotaxane.
[25] The reactive group added to both ends of the linear polymer portion is selected from the group consisting of an amino group, a carboxyl group, an aldehyde group, a sulfanyl group, an azide group, an alkynyl group, a tosyl group, and an active ester group. The method according to [24].

According to one aspect of the present invention, at least two lines comprising a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymers are linked via at least one degradable moiety. Polyrotaxane compound including a linear polymer portion, in other words, a polyrotaxane compound including a plurality of cyclic molecules and one linear polymer having a terminal group, and having at least one decomposable portion inside the linear polymer Is provided.

Since the degradable polyrotaxane compound according to the present invention has a degradable portion inside the linear polymer main chain, it is decomposed compared to a conventional degradable polyrotaxane compound having degradable bonds at both ends. Larger changes in molecular weight can occur at times, which can dramatically change the basic physical properties of polymers such as viscosity, solubility, and glass transition point. In addition, it is fundamentally different from conventional double-end-decomposable polyrotaxanes, and molecules with a bulky structure such as enzymes are arranged because degradable bonds are placed inside the linear polymer main chain at the end away from the terminal site. It is easy to access the degradable part of the material, and it can be expected to design a wide range of decomposition responsiveness and improve the decomposition efficiency. Furthermore, since the conventional degradable functional polyrotaxane introduces degradable bonds at both ends of the linear polymer chain, it is necessary to arrange two degradable bonds with respect to one molecule of polyrotaxane. Required more than 6-7 steps. On the other hand, in the degradable functional polyrotaxane according to the present invention, the degradable bond is arranged only near the center of the linear polymer, so that the synthesis method is simplified within 4 steps, and the production time is shortened. There is also an advantage that an improvement in the recovery amount can be achieved.

FIG. 3 is a diagram showing a ¹ H-NMR spectrum of polyrotaxane A having a decomposable group. It is the figure which showed the GPC chart of the polyrotaxane A after ultraviolet irradiation. FIG. 3 is a diagram showing a ¹ H-NMR spectrum of polyrotaxane B having a methacryloyl group as a polymerizable group. It is the figure which showed the result of the tension test of the Bis-GMA containing hardening body which does not contain a polyrotaxane. It is the figure which showed the result of the tensile test of the hardening body containing polyrotaxane B. Polyrotaxane, 2-dimethylaminoethyl methacrylate, camphorquinone, 2-hydroxyethyl methacrylate are mixed in a silicone mold (thickness 1 mm, length 15 mm, dumbbell shape with a center width 1 mm, end width 2 mm). It is the figure shown about producing a hardening body by adding the photopolymerization type adhesive agent prepared in this way. It is the graph which showed the micro tensile strength of the non-UV irradiation group and the UV irradiation group about the polyrotaxane of a different mass part.

One aspect of the present invention includes a plurality of cyclic molecules and one linear polymer having a terminal group, and the linear polymer is linked via at least one degradable moiety. A polyrotaxane compound containing a linear polymer portion, in other words, a plurality of cyclic molecules and one linear polymer having a terminal group, and at least one decomposable inside the linear polymer away from the terminal The present invention relates to a polyrotaxane compound having a moiety. The present invention is described in detail below.

Polyrotaxane (PRX) compound Rotaxane is a macromolecule that is penetrated by a macromolecule and bonds bulky sites to both ends of the macromolecule so that the ring cannot be removed from the shaft due to steric hindrance. . In polyrotaxane, one linear polymer penetrates the ring of a plurality of macrocyclic molecules. As the linear polymer and cyclic molecule used in the present invention, known molecules can be used, and are not particularly limited.

The linear polymer contained in the polyrotaxane according to one embodiment of the present invention has a structure represented by the following formula:
X ¹ -Y ⁰ (-Z ⁱ -Y ⁱ ) _i -X ²
Here, ^{X 1} and ^{X 2} are the same or different terminal group, ^{Y 0} and ^{Y i} are identical or different linear polymeric moiety, ^{Z i} is an exploded moiety, ^(- Z ⁱ -Y i I) _i indicates that there are i repeating units composed of a decomposable portion Y and a linear polymer portion Z, and each Z ⁱ and Y ⁱ in the linear polymer may be the same or different. Although i is not particularly limited, i is preferably an integer of 1 to 500, more preferably 1 to 10, for example 1 or 2.

For example, when i = 1, the linear polymer has a structure in which two linear polymer parts are linked via one degradable part as shown in the following formula:
X ¹ -Y ⁰ -Z ¹ -Y ¹ -X ²
Here, X ¹ and X ² are the same or different end groups, Y ⁰ and Y ¹ are the same or different linear polymer moieties, and Z ¹ is a degradable moiety.

For example, when i = 2, the linear polymer has a structure in which three linear polymer parts are connected via two degradable parts as shown in the following formula:
X ¹ -Y ⁰ -Z ¹ -Y ¹ -Z ² -Y ² -X ²
Here, X ¹ and X ² are the same or different end groups, Y ⁰ , Y ¹ and Y ² are the same or different linear polymer moieties, and Z ¹ and Z ² are the same or different degradable moieties. is there.

As the linear polymer or the linear polymer portion, a known polyrotaxane can be appropriately selected. For example, polyethylene glycol, polypropylene glycol, a copolymer of polyethylene glycol and polypropylene glycol (poloxamer), polyisoprene. , Polyisobutylene, polyisobutene, polybutadiene, polytetrahydrofuran, polyacrylic ester, polydimethylsiloxane, polyethylene, polymethyl vinyl ether, polypropylene, polyperfluorooxypropylene, oligotetrafluoroethylene, polycaprolactam, polyethyleneimine, polyamino acid, and polymethyl It is preferably selected from the group consisting of vinyl ethers. The average molecular weight of the linear polymer is not particularly limited, but is preferably 1000 to 100,000, particularly 2000 to 40000 or 5000 to 20000. Two or more linear polymer portions contained in one linear polymer may be the same as or different from each other. The average molecular weight of the linear polymer portion is not particularly limited, but is preferably 100 to 100,000, particularly 200 to 40,000 or 500 to 20,000.

As the cyclic molecule, a conventionally known cyclic molecule can be used. α, β or γ-cyclodextrin is preferred, but it may have a cyclic structure similar to this, and examples of such cyclic structure include cyclic polyether, cyclic polyester, cyclic polyetheramine, cyclic Polyamine, cyclophane, crown ether and the like can be mentioned. For example, a preferable cyclic molecule from the viewpoint of the ability to form pseudopolyrotaxane is α, β or γ-cyclodextrin, and α-cyclodextrin is particularly preferable.

The cyclic molecule contained in the polyrotaxane according to the present invention may have one or a plurality of substituents. When cyclodextrin is used as the cyclic molecule, a substituent can be introduced into the hydroxyl group of cyclodextrin. Examples of the substituent include 2-hydroxyethoxyethyl (HEE) group, hydroxyethyl group, hydroxypropyl group, hydroxybutyl group, hydroxyethoxyethyl group, N, N-dimethylaminoethyl group (sometimes referred to as DMAE group). ), Carboxyl group, methyl group, sulfate group (sulfo group), primary amino group, or water-soluble polymer such as polyethylene glycol, protein molecule such as collagen or transferrin, peptide molecule including RGD motif or peptide molecule such as oligoarginine And polymerizable groups used for cross-linking between molecules. The cyclic molecule may contain multiple types of substituents. Examples of the polymerizable group include (meth) acryloyl group, (meth) acryloyloxy group, (meth) acryloylamino group, (meth) acryloyl group derivative group such as (meth) acryloylthio group, vinyl group, aryl group, styryl. Group, (meth) acrylamide group, etc. can be used suitably. A preferred polymerizable group is a methacryloyl group. The polyrotaxane compound according to the present invention may contain a plurality of types of polymerizable groups. These groups may be directly bonded to the cyclic molecule or may be bonded via a linker. The linker is not particularly limited and may be appropriately selected depending on the intended purpose. For example, the linker may be a carbamate ester bond (—O—CO—NH—), an ester bond (—O—CO—), a carbonate bond (— O-CO-O-), ether bond (-O-) and the like.

As the combination of the linear polymer and the cyclic molecule, a combination of α-cyclodextrin and polyethylene glycol, a combination of β-cyclodextrin and poloxamer, or the like is preferable. The synthesis of polyrotaxane by a combination of β-cyclodextrin and poloxamer is also disclosed in Patent Document 1 described above, the contents of which are also incorporated herein by reference. The ratio between the number of molecules of the linear polymer and the number of molecules of the cyclic molecule is not particularly limited, but is preferably 1: 1 to 1: 500, more preferably 1: 5 to 1: 200. A ratio of 1:10 to 1: 100 is used. That is, preferably 1 to 500 cyclic molecules are contained in one molecule of the linear polymer, more preferably 5 to 200 cyclic molecules, for example, 10 to 100 cyclic molecules may be contained.

Examples of the terminal group (also referred to as bulky substituent) used in the present invention include 2,4-dinitrophenyl group, 3,5-dinitrophenyl group, cyclodextrin, adamantane group, O-triphenylmethyl (O— Trt) group, S-triphenylmethyl (S-Trt) group, N-triphenylmethyl (N-Trt) group, N-tritylglycine, benzyloxycarbonyl (Z) group, 9-fluorenylmethyloxycarbonyl ( Fmoc) group, benzyl ester (OBz) group, tertiary butylcarbonyl (Boc) group, amino acid tertiary butyl ester (OBu group), trityl group, fluorescein, pyrene, substituted benzene (as substituents, alkyl, alkyloxy, hydroxy) , Halogen, cyano, sulfonyl, carboxyl, amino, phenyl, etc. A polynuclear aromatic which may be substituted (but not limited to these), and may be the same as those described above, but is not limited thereto. One or more substituents may be present. May be selected from the group consisting of, but not limited to, MPC (2-methacryloyloxyethyl phosphorylcholine), BMA (n-butyl methacrylate), a combination of MPC and BMA, and steroids. One preferred end group is an adamantane group. The end group need not be directly linked to the linear polymer moiety, but via a linker moiety known to those skilled in the art (eg, a moiety containing a peptide bond, carbamate bond, ester bond, or ether bond). It may be connected.

The polyrotaxane compound according to the present invention includes a plurality of cyclic molecules and one linear polymer having a terminal group, and has at least one decomposable portion inside the linear polymer away from the terminal. The degradable moiety of the polyrotaxane compound according to the present invention includes at least one degradable group. Specific examples of the decomposable group include p-methoxyphenacyl group, 2-nitrobenzyl group, 2-nitrobenzyloxycarbonyl group, 2-nitrophenylethylene glycol group, benzyloxycarbonyl group, 3,5-dimethoxybenzyloxy Carbonyl group, α, α-dimethyl-3,5-dimethoxybenzyloxycarbonyl group, 3-nitrophenyl group, 3-nitrophenoxy group, 3,5-dinitrophenoxy group, 3-nitrophenoxycarbonyl group, phenacyl group, 4 -Methoxyphenacyl group, α-methylphenacyl group, 3,5-dimethoxybenzoinyl group, 2,4-dinitrobenzenesulfenyl group, (coumarin-4-yl) methyl group, 7-nitroindolinyl group, Photocleavable groups such as arylazophosphate ester units; ester bonds, Schiff bases Various hydrolyzable bonds such as carbamate bonds, peptide bonds, ether bonds, acetal bonds and hemiacetal bonds; bonds decomposable by reducing agents such as disulfide bonds; thermal decomposable groups such as organic peroxides; An acyl hydrazine bond etc. are mentioned. These may be used individually by 1 type and may use 2 or more types together. Preferable degradable groups include, for example, a photocleavable group, a disulfide group, and a peptide consisting of 2 to 30 amino acid residues. Peptides may have specific sequences that are recognized and cleaved by enzymes such as specific proteases and peptidases.

When the linear polymer has only one degradable portion, the position is preferably near the center of the linear polymer, in other words, two linear shapes connected to the degradable portion. The length ratio of the polymer portion is preferably 1: 1, but the position is not limited. The ratio of the lengths of the two linear polymer portions linked to the degradable portion can be any ratio included, for example, from 1: 1 to 1: 4. For example, when the degradable part is located in the center of the linear polymer, two linear polymer parts having a length half that of the linear polymer are generated along with the decomposition of the decomposable part. Cyclic molecules are released from the non-existing ends. When the linear polymer has a plurality of degradable portions, the ratio of the lengths of the linear polymer portions connected to each degradable portion is also preferably 1: 1, but there is no particular limitation. It can be any ratio comprised between 1: 1 and 1: 4.

The number average molecular weight of the polyrotaxane according to the present invention is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably about 10,000 to 500,000.

Dental Material Composition / Adhesive Composition The polyrotaxane compound according to the present invention can be used as a dental adhesive. Since the photodegradable polyrotaxane having a polymerizable functional group introduced into a plurality of cyclodextrins (CD) functions as a crosslinking agent, a three-dimensional structure can be easily produced by copolymerizing with other monomers. Furthermore, the three-dimensional structure can be decomposed by irradiation with ultraviolet rays to reduce the mechanical strength. For example, when such a photodegradable polyrotaxane crosslinking agent is used as a resin monomer in a dental material, it can be developed into a dental adhesive that is cured by irradiation with visible light and decomposed by irradiation with ultraviolet light. For example, Seo et al. Reported the construction of a three-dimensional structure using a photodegradable polyrotaxane in which nitrobenzyl, which is photolyzed by ultraviolet irradiation, is arranged between both ends of a linear polymer and a blocking group (Seo). et al., ACS Macro Lett., 4, 1154, 2015). Therefore, one embodiment of the present invention includes a plurality of cyclic molecules and one linear polymer having a terminal group, and the linear polymer is linked via at least one degradable moiety. The present invention relates to an adhesive composition containing a polyrotaxane compound containing at least two linear polymer moieties. Another embodiment of the present invention includes a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymer is linked via at least one degradable moiety. The present invention relates to a dental material composition containing a polyrotaxane compound containing two linear polymer parts.

The above composition according to the present invention may further contain a polymerizable monomer. The polyrotaxane compound contained in the composition according to the present invention is preferably soluble in a solution of a polymerizable monomer. As the polymerizable monomer, for example, those having at least one polymerizable group in the molecule such as a polymerizable unsaturated group, a ring-opening polymerizable group, and a polycondensable group can be used. Conventionally known polymerizable monomers used in known adhesive compositions can be used without limitation. These polymerizable monomers cause a polymerization initiation reaction by the action of heat, a polymerization initiator, gamma rays, electrolysis, plasma, and the like. Examples of the polymerizable monomer include 2-ethylhexyl (meth) acrylate, glycidyl (meth) acrylate, benzyl (meth) acrylate, ethylene glycol di (meth) acrylate, diethylene glycol di (meth) acrylate, triethylene glycol di ( And (meth) acrylate, methyl (meth) acrylate, 4-methacryloxyethyl trimellitate anhydride and the like.

The composition according to the present invention may further contain a polymerization initiator. As the polymerization initiator, conventionally known polymerization initiators used for conventionally known radical polymerization, anionic polymerization, cationic polymerization, polyaddition reaction, polycondensation reaction, coupling reaction, inorganic synthetic polymer synthesis and the like can be used. For example, radical polymerization initiators used for photopolymerization include benzoin methyl ether, benzyl dimethyl ketal, benzophenone, 4,4′-dimethylbenzophenone, diacetyl, 2,3-pentadione benzyl, camphorquinone, 9,10-phenone. Examples thereof include nantraquinone and 9,10-anthraquinone.

Surface Coating Agent / Adhesion Prevention Agent The polyrotaxane compound according to the present invention can be used as a coating agent on the surface of a material. For example, by modifying various functional groups and cell-adhesive oligopeptides to CD, Seo et al. Can control protein-level and cell-level biointerfaces such as inactive surface adsorption of proteins and rapid cell adhesion. (J. Am. Chem. Soc., 2013, 135, 5513). Furthermore, Seo et al. Analyzed, for example, the adsorption of fibrinogen that activates platelets using a polyrotaxane surface obtained by methylating CD. As a result, fibrinogen adsorbed on the surface has a suppressed conformational change and is in an inactive state. It has also been reported (Soft Matter, 2012, 8, 5477). It is known that a three-dimensional structure produced using a polyrotaxane has mechanical properties such as collagen fibers constituting skin and blood vessels. That is, it exhibits flexibility for small deformations and high rigidity for large deformations (see, for example, Ito, Polym. J., 2007, 39, 489). Materials with such properties can follow the movements of the body, such as stretching and contracting of muscles and tendons, contraction of the stomach and heart beat, and adhere to tissue composed of cells and extracellular matrix. Since it can be controlled, it can be developed as a wearable device, a wound healing material, or an anti-adhesion agent. Therefore, one embodiment of the present invention includes a plurality of cyclic molecules and one linear polymer having a terminal group, and the linear polymer is linked via at least one degradable moiety. The present invention relates to a surface coating agent containing a polyrotaxane compound containing at least two linear polymer moieties. Another embodiment of the present invention includes a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymer is linked via at least one degradable moiety. The present invention relates to an anti-adhesion agent containing a polyrotaxane compound containing two linear polymer moieties.

In-vivo implant / tissue regenerator The polyrotaxane compound according to the present invention can be used as an in-vivo implant or tissue regenerative device by using a three-dimensional structure produced by crosslinking or the like. For example, in a previous study based on polyrotaxane, Seo et al. Can regulate the formation of cytoskeletal proteins by controlling the number of cyclodextrin (CD) penetrations, and surface with low surface molecular mobility. The upper mesenchymal stem cells promote the formation of actin fibers, which are cytoskeletal proteins, and differentiate into osteoblasts. On surfaces with high surface molecular mobility, the formation of actin fibers is inhibited and differentiates into adipocytes. (Adv. Healthcare Mater., 2015, 4, 215). Since materials having such properties can be rapidly decomposed and lost after a certain period of time when cell differentiation and proliferation are promoted or suppressed in vivo, surface treatment of various implantable medical devices, It can be developed as various devices for cell culture used in vivo or in vitro for the purpose of implants and tissue regeneration.

In cells that adhere to culture equipment, it is known that a physiological activity signal is transmitted into the cells reflecting the physicochemical characteristics of the equipment, thereby regulating metabolic activities such as cell proliferation, differentiation, and death. . For this reason, research on physicochemical factors on the surface of materials and control of cell functions according to them has been actively conducted for the past decade. For example, the differentiation characteristics of mesenchymal stem cells on elastomeric surfaces with various elasticity are enhanced by the differentiation of nerves and adipocytes being promoted by lowering the Young's modulus of the adhered elastomer. It is known that differentiation into osteoblasts is promoted.

In addition, human embryonic stem cells (hESC) are sensitive to physical stimuli. When the matrix stiffness of the micropost array on the incubator increases, the cytoskeletal contractility increases, and the rigid substrate maintains hESC pluripotency. It is known to be promoted.

As a technology to control the differentiation or undifferentiation maintenance of stem cells by adjusting the physicochemical properties of such equipment, pluripotent stem cells are cultured using incubators with polyrotaxane block copolymer surfaces with different molecular motility. It is possible to maintain the undifferentiated state by culturing or to induce differentiation into specific cells.

Even if control of cell differentiation or undifferentiation maintenance is achieved in vivo or in vitro by adjusting the physicochemical characteristics of the equipment as described above, the cells or formed tissue can be used in vivo. Need to be removed. In addition, in the case of regeneration at a tissue defect site in a living body, since it is necessary to regenerate the tissue of the site while maintaining the space of the defect site by using scaffold implantation, Biodegradation is essential (Non-Patent Documents 1 and 2).

The implant material and cell culture device according to the present invention can be produced by crosslinking a degradable polyrotaxane. A decomposable polyrotaxane is prepared by introducing a functional group that forms a crosslinking point into a linear polymer terminal or in the vicinity of the terminal, or in a cyclic molecule, and can be prepared by crosslinking it alone or with other reactive molecules. Examples of the crosslinkable functional group include a vinyl group, an aldehyde group, and a carboxyl group. Examples of the reactive molecule used for the crosslinking reaction include vinyl polymerizable monomers, polysaccharides, proteins (polypeptides) and the like.

Scaffolds composed of degradable polyrotaxanes are capable of degrading and disappearing after a certain period of time when cell differentiation and proliferation were promoted in vivo by the molecular mobility of polyrotaxanes, and cells or tissues can be integrated with surrounding tissues. Thus, it is possible to secure the space of the defect site at 1), to regenerate the tissue at the defect site, and to remove the scaffold after regeneration.

Therefore, one embodiment of the present invention includes a plurality of cyclic molecules and one linear polymer having a terminal group, and the linear polymer is linked via at least one degradable moiety. The present invention relates to an in vivo implant containing a polyrotaxane compound containing at least two linear polymer moieties. Another embodiment of the present invention includes a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymer is linked via at least one degradable moiety. The present invention relates to a tissue regeneration device containing a polyrotaxane compound containing two linear polymer portions. These implants and tissue regenerators may contain other materials such as collagen and gelatin, preferably bioabsorbable materials.

Pharmaceutical composition The polyrotaxane compound according to the present invention is used for the treatment of diseases caused by abnormal cell metabolic functions such as lysosomal disease, diseases caused by intracellular cholesterol accumulation, or diseases caused by dysfunction of autophagy. It can be used as an active ingredient of a pharmaceutical composition. As the degradable portion of the polyrotaxane compound according to the present invention, a degradable group capable of degrading in an acidic environment within a cell and in an acidic environment at pH 4.0 to 6.0 can be employed. Such a polyrotaxane compound is decomposed in an acidic environment, the polyrotaxane skeleton is collapsed, and a cyclic molecule such as β-CD is released.

It is known that vesicles such as lysosomes and late endosomes exist in eukaryotic cells including humans, and the lumens of these vesicles are acidified. For example, the pH of the lysosomal lumen is around 5. Therefore, the polyrotaxane compound according to the present invention can be decomposed by being taken into these vesicles. The polyrotaxane compound according to the present invention releases a cyclic molecule such as β-CD upon decomposition. For example, when β-cyclodextrin is released in lysosomes, it can include cholesterol present in lysosomes, thereby causing Niemann-Pick disease type C, which is caused by excessive accumulation of cholesterol in lysosomes. Lysosomal disease can be treated or prevented (see, eg, Tamura and Yui, Sci. Rep., 2014, 4, 4356).

It will be understood by those skilled in the art that inclusion of β-cyclodextrin in intracellular cholesterol enables treatment of diseases caused by excessive intracellular cholesterol. Therefore, one embodiment of the present invention relates to a pharmaceutical composition for treating a disease caused by intracellular cholesterol accumulation. In another aspect, one embodiment of the present invention relates to a method for treating or preventing Niemann-Pick disease type C (NPC). Furthermore, one aspect of the present invention also relates to the use of a polyrotaxane compound in the manufacture of a medicament for treating or preventing Niemann-Pick disease type C (NPC). Preferred acid-decomposable polyrotaxane compounds as pharmaceutical compositions include, for example, compounds having a disulfide bond in the interior of the linear polymer, preferably the central portion, and an N-triphenylmethyl (N-Trt) group at the terminal. included.

Diseases resulting from accumulation of metabolites in lysosomes include lysosomal disease, more specifically Gaucher disease (Gaucher disease), Neimann-Pick disease type A (Niemann-Pick disease type A), Neiman-Pick disease. B type (Niemann-Pick disease type B), Niemann-Pick disease type C (Niemann-Pick disease type C), GM1 gangliosidosis, GM2 gangliosidosis ("Tay-Sachs Sandhoff type AB"). ), Krabbe disease (Krabe disease), metachromatic leukodystrophy, multiple sulphafase deficiency (Multisulfatese deficiency), Faber disease (Farber disease), mucopolysaccharidosis type I, mucopolysaccharidosis type II ("Hunter Sometimes referred to as "disease"), mucopolysaccharidosis I Type I (sometimes referred to as “Sanfiliposis”), mucopolysaccharidosis type IV, mucopolysaccharidosis type VI (sometimes referred to as “Maloto Lamy disease”), mucopolysaccharidosis type VII (“Sly disease”) ), Mucopolysaccharidosis type IX (sometimes referred to as “Hyaluronidase deficiency”), sialidosis, galactosialidosis, I-cell disease / mucolipidosis type III, α-mannosidosis, β-mannosidosis, fucosidosis, aspartylglucosamineuria, Schindler / Kanzaki disease (Schindler / Kanzaki disease), Wolman disease (Wolman disease), Danone disease (Danon disease), free sialic acid storage disease, ceroid lipofusti Examples include nosis and Fabry disease. The lysosomal disease is also a disease caused by autophagy dysfunction causing autophagosome accumulation.

Therefore, one embodiment of the present invention relates to a pharmaceutical composition for treating or preventing Niemann-Pick disease type C (NPC). In another aspect, one embodiment of the present invention relates to a method for treating or preventing Niemann-Pick disease type C (NPC). Furthermore, one aspect of the present invention also relates to the use of a polyrotaxane compound in the manufacture of a medicament for treating or preventing Niemann-Pick disease type C (NPC).

The polyrotaxane compound according to the present invention can be used as an active ingredient of a composition for use in inducing autophagy in cells. Accordingly, one embodiment of the present invention relates to a composition for inducing autophagy in a cell. In another aspect, one embodiment of the present invention relates to a method for inducing autophagy in a cell. Furthermore, one aspect of the present invention also relates to the use of a polyrotaxane compound in the manufacture of a medicament for inducing autophagy in a cell.

The methylated polyrotaxane according to the present invention can induce autophagic cell death in cells. It is known to those skilled in the art that autophagic cell death can be used to induce cell death in cancer cells. Thus, one aspect of the present invention relates to a pharmaceutical composition for treating cancer, preferably a pharmaceutical composition for treating cancer resistant to apoptosis. From another aspect, one embodiment of the present invention relates to a method for treating cancer, particularly for treating cancer resistant to apoptosis. Furthermore, one aspect of the present invention pertains to the use of polyrotaxane compounds in the manufacture of a medicament for treating cancer.

Further, one embodiment of the present invention relates to a pharmaceutical composition for treating or preventing a disease caused by autophagy dysfunction. In another aspect, one embodiment of the present invention relates to a method for treating or preventing a disease caused by a dysfunction of autophagy. Furthermore, one aspect of the present invention also relates to the use of a polyrotaxane compound in the manufacture of a medicament for treating or preventing a disease resulting from autophagy dysfunction. Examples of diseases caused by autophagy dysfunction include the above-mentioned lysosomal diseases, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and Huntington's disease.

The polyrotaxane compound according to the present invention can be used as an active ingredient in a pharmaceutical composition used for the treatment or prevention of the above-mentioned diseases. Therefore, one embodiment of the present invention relates to a pharmaceutical composition used for treatment or prevention of diseases. The other components in the pharmaceutical composition according to the present invention are not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include pharmaceutically acceptable carriers. There is no restriction | limiting in particular also in a support | carrier, For example, it can select suitably according to a dosage form etc. There is no restriction | limiting in particular also about content in the pharmaceutical composition based on this invention, According to the objective, it can select suitably. Preferably, the pharmaceutical composition according to the present invention is water-soluble at around body temperature, for example, 34 ° C to 42 ° C, more preferably 35 ° C to 38 ° C or 37 ° C.

There is no restriction | limiting in particular as a dosage form of the pharmaceutical composition based on this invention, According to the desired administration method, it can select suitably, For example, an injection (Solution, suspension, solid agent for use, etc.) And inhaled powders. As an injection, for example, a pH regulator, a buffer, a stabilizer, a tonicity agent, a local anesthetic, etc. are added to the polyrotaxane compound according to the present invention, and subcutaneous, intramuscular, intravenous, etc. are added by a conventional method. An injection for internal use can be produced. Examples of the pH adjusting agent and the buffering agent include sodium citrate, sodium acetate, sodium phosphate and the like. Examples of the stabilizer include sodium pyrosulfite, EDTA, thioglycolic acid, thiolactic acid and the like. Examples of the isotonic agent include sodium chloride and glucose. Examples of the local anesthetic include procaine hydrochloride and lidocaine hydrochloride.

The administration method of the pharmaceutical composition according to the present invention is not particularly limited, and for example, either local administration or systemic administration can be selected according to the dosage form of the pharmaceutical composition, the patient's condition, and the like. For example, local administration includes intracerebroventricular administration.

The subject of administration of the pharmaceutical composition according to the present invention is not particularly limited and may be appropriately selected depending on the purpose. Examples thereof include humans, mice, rats, cows, pigs, monkeys, dogs, cats and the like. However, it is preferably a human.

The dosage of the pharmaceutical composition according to the present invention is not particularly limited, and can be appropriately selected depending on the dosage form, the age and weight of the administration subject, the degree of desired effect, and the like.

The administration time of the pharmaceutical composition according to the present invention is not particularly limited and may be appropriately selected depending on the purpose. For example, it may be administered prophylactically to a patient susceptible to the above-mentioned diseases, It may be administered therapeutically to patients presenting with symptoms. Moreover, there is no restriction | limiting in particular as frequency of administration, According to the age of an administration subject, a body weight, the grade of a desired effect, etc., it can select suitably.

Production Method The degradable polyrotaxane compound according to the present invention can be synthesized more easily and in a shorter time than a conventional degradable polyrotaxane compound having degradable groups at both ends. One embodiment of the present invention includes a plurality of cyclic molecules and one linear polymer having a terminal group, and the linear polymer is connected via at least one degradable moiety. A method for producing a polyrotaxane compound comprising a polymer portion, wherein a) a step of adding a reactive group to both ends of the linear polymer portion, b) a linear polymer portion having reactive groups added to both ends To obtain a linear polymer comprising at least two linear polymer moieties linked via at least one degradable moiety, and c) the linear polymer as a cyclic molecule. To a pseudopolyrotaxane, and d) adding a terminal group to both ends of the pseudopolyrotaxane.

In the above step a, the reactive group added to both ends of the linear polymer portion is, for example, a leaving group such as an amino group, a carboxyl group, an aldehyde group, a sulfanyl group, an azido group, an alkynyl group, a tosyl group, It may be an active ester group such as a carboxylic acid succinimide ester.

After the step b, the linear polymer may be selected based on the length of the linear polymer. Preferably, a linear polymer in which two linear polymer parts are connected via one degradable part is selected.

Hereinafter, the present invention will be specifically described with reference to examples. However, the present invention is not limited to these examples.

Example 1 Synthesis of Centrally Decomposable Polyrotaxane Crosslinking Agent [Synthesis Example 1]
<Synthesis of polyrotaxane A having a decomposable group in the molecule of the linear shaft polymer>
(1) Activation of both ends of polyethylene glycol with carbodiimidazole (Preparation of PEG5k-CDI)
A solution obtained by dissolving 20.0 g of polyethylene glycol (PEG5k-OH) (weight average molecular weight: 4,400-4,800) in 240 mL of tetrahydrofuran in 10.8 g of 1,1-carbonyldiimidazole (CDI) under a nitrogen atmosphere The solution was slowly added dropwise at room temperature. After completion of the dropwise addition, the mixture was stirred at room temperature for 24 hours under a nitrogen atmosphere. The reaction solution was concentrated and then added dropwise to diethyl ether to collect the precipitate, which was dried (collected amount 19.9 g).

(2) Amination of both ends of polyethylene glycol (Preparation of PEG5k-NH ₂ )
19.9 g of PEG5k-CDI obtained in (1) above was completely dissolved in 240 mL of tetrahydrofuran, and then slowly added dropwise to 28.5 mL of ethylenediamine at room temperature in a nitrogen atmosphere. After completion of the dropwise addition, the mixture was stirred at room temperature under a nitrogen atmosphere for 24 hours. The reaction solution was concentrated and then dropped into diethyl ether to collect the precipitate. The obtained precipitate was dialyzed against ultrapure water for 2 days using a dialysis membrane (molecular weight cut off: 500), and then freeze-dried and recovered as a solid (recovered amount 16.42 g).

(3) Introduction of degradable group into aminated polyethylene glycol (Preparation of cNBPEG10k-NH ₂ )
15.0 g of PEG5k-NH ₂ obtained in the above (2) was completely dissolved in 45 mL of tetrahydrofuran. Also, using another glass container, 137.4 mg of CDI and 2-nitro-p-xylene glycol were dissolved in 10 mL of tetrahydrofuran and reacted at room temperature for 2 hours, and then this reaction solution was converted into a PEG5k-NH ₂ / terolahydrofuran solution. It was dripped. After completion of the dropwise addition, the mixture was stirred at room temperature under a nitrogen atmosphere for 24 hours. The reaction solution was dropped into diethyl ether, and the precipitate was collected. The resulting precipitate was dialyzed against ultrapure water using a dialysis membrane (fractionated molecular weight 3,500) for 5 days, and then freeze-dried and recovered as a solid (recovered amount 7.4 g).

(4) cNBPEG10k-NH ₂ and of Inclusion Complex by using α- cyclodextrin preparation and blockade of inclusion complexes (prepared polyrotaxane A having decomposable group in the molecule of the linear axes polymers)
15 g of α-cyclodextrin (α-CD) was dissolved in 103 mL of ultrapure water, and a solution of 3 g of cNBPEG10k-NH ₂ obtained in (3) above was dissolved in 12 mL of ultrapure water. After stirring for 24 hours at room temperature, the clathrate complex precipitated as turbid was lyophilized and recovered.

A solution obtained by dissolving 4.0 g of BOP reagent, 2.2 g of 1-adamantanecarboxylic acid and 2.1 mL of N, N-diisopropylethylamine in 120 mL of N, N-dimethylformamide and the recovered inclusion complex were mixed at room temperature. The reaction was performed for 24 hours.

The precipitate formed by dropping the reaction solution into methanol was collected by centrifugation. The operation of dissolving the collected precipitate in dimethyl sulfoxide and precipitating with methanol was repeated several times. Next, the operation of dissolving the collected precipitate in dimethyl sulfoxide and precipitating with water was repeated several times. After recovery by centrifugation, lyophilization yielded a purified polyrotaxane (polyrotaxane A having a degradable group) (recovery amount 9.2 g).

The obtained polyrotaxane (polyrotaxane A having a decomposable group) was identified by ¹ H-NMR and GPC, and it was confirmed that unencapsulated CD was not contained. FIG. 1 shows the ¹ H-NMR spectrum of Polyrotaxane A. Further, when the number of penetrations of α-CD with respect to PEG was calculated, the number of penetrations of α-CD was 28.1 molecules.

Polyrotaxane A was dissolved in 1 mL of dimethyl sulfoxide. Thereafter, ultraviolet rays (254 nm, 2.5 mW / cm ² ) were irradiated for 1, 5 and 10 minutes, and it was confirmed that the molecular weight was reduced by gel permeation chromatography (GPC). FIG. 2 shows a GPC chart of Polyrotaxane A after ultraviolet irradiation. This result indicates that the axial polymer chain is cleaved by cleavage of nitrobenzene having photodegradability, and a part or all of α-CD is released from the inclusion state to the non-inclusion state. That is, it was confirmed that the supramolecular structure of polyrotaxane A was decomposed and collapsed by irradiation with ultraviolet rays (254 nm, 2.5 mW / cm ² ).

[Synthesis Example 2]
<Synthesis of Polyrotaxane A Having Methacryloyl Group as Hydrophobic Group and Polymerizable Group (Preparation of Polyrotaxane B)>
(5) Polymerization and hydrophobization of polyrotaxane A having a decomposable group (Preparation of polyrotaxane B)
1 g of “polyrotaxane A having a decomposable group” synthesized in the above (4) was dissolved in 15 mL of dehydrated dimethyl sulfoxide, and 209.9 μL of 2-isocyanatoethyl methacrylate and 1.3 mL of butyl isocyanate were added. 1,4-diazabicyclo [2.2.2] octane (500 mg) was added, and the mixture was reacted at room temperature for 24 hours. After the reaction, dialysis was carried out for 2 days against dimethyl sulfoxide using a dialysis membrane (fraction molecular weight 3,500). Subsequently, dialysis was performed for 2 days against water using the same dialysis membrane (fraction molecular weight 3,500). From the results of 1H-NMR measurement, 1.2 molecules of methacryloyl group and 8.5 molecules of butyl isocyanate per α-CD molecule included from the peak derived from α-CD, the peak derived from aminobutyl group, and the peak derived from methacryloyl group It was confirmed that the group was introduced. FIG. 3 shows the 1H-NMR spectrum of Polyrotaxane B. The molecular weight was confirmed by GPC. Moreover, as a result of confirming an absorption spectrum, it confirmed that polyrotaxane B had the absorption derived from a nitrobenzyl group. That is, it was confirmed that polyrotaxane B was a predetermined polyrotaxane B having a nitrobenzyl group as a photocleavable group as a decomposable group and a methacryloyl group as a polymerizable group.

[Comparative Synthesis Example 1]
<Synthesis of polyrotaxane having no degradable group>
(6) Activation of both ends of polyethylene glycol with carbodiimidazole (Preparation of PEG5k-CDI)
A solution prepared by dissolving 10.0 g of polyethylene glycol (PEG10k-OH) (weight average molecular weight: 10,000) in 2.4 mL of 1,1-carbonyldiimidazole (CDI) and 100 mL of tetrahydrofuran was slowly added at room temperature under a nitrogen atmosphere. And dripped. After completion of the dropwise addition, the mixture was stirred at room temperature for 24 hours under a nitrogen atmosphere. The reaction solution was concentrated and then dropped into diethyl ether, and the precipitate was collected and dried (collected amount 9.6 g).
(7) Amination of both ends of polyethylene glycol (Preparation of PEG10k-NH ₂ )
After 9.5 g of PEG10k-CDI obtained in (6) above was completely dissolved in 100 mL of tetrahydrofuran, it was slowly added dropwise to 6.3 mL of ethylenediamine at room temperature under a nitrogen atmosphere. After completion of the dropwise addition, the mixture was stirred at room temperature under a nitrogen atmosphere for 24 hours. The reaction solution was concentrated and then dropped into diethyl ether to collect the precipitate. The obtained precipitate was dialyzed against ultrapure water using a dialysis membrane (fractionated molecular weight 3,500) for 2 days, and then freeze-dried and recovered as a solid (recovered amount 7.5 g).

(8) Preparation of inclusion complex using PEG10k-NH ₂ and α-cyclodextrin and blocking of the inclusion complex (preparation of polyrotaxane X having no decomposable group in the molecule of the linear shaft polymer)
15 g of α-cyclodextrin (α-CD) was dissolved in 103 mL of ultrapure water, and a solution prepared by dissolving 3 g of PEG10k-NH ₂ obtained in (7) above in 12 mL of ultrapure water was added dropwise thereto. After stirring for 24 hours at room temperature, the clathrate complex precipitated as turbid was lyophilized and recovered. A solution obtained by dissolving 4.0 g of BOP reagent, 2.2 g of 1-adamantanecarboxylic acid and 2.1 mL of N, N-diisopropylethylamine in 120 mL of N, N-dimethylformamide and the recovered inclusion complex were mixed at room temperature. The reaction was performed for 24 hours.

A precipitate formed by dropping the reaction solution into methanol was collected by centrifugation. The operation of dissolving the collected precipitate in dimethyl sulfoxide and precipitating with methanol was repeated several times. The operation of dissolving the collected precipitate in dimethyl sulfoxide and precipitating with water was repeated several times. After recovery by centrifugation, lyophilization yielded a purified polyrotaxane (polyrotaxane X having no degradable group) (recovered amount 10.1 g). The obtained polyrotaxane (polyrotaxane X having no decomposable group) was identified by ¹ H-NMR and GPC, and it was confirmed that unencapsulated CD was not contained. Further, when the number of penetrating α-CD to PEG was calculated, the number of penetrating α-CD was 28.8 molecules. Polyrotaxane X was dissolved in 1 mL of dimethyl sulfoxide. Thereafter, ultraviolet rays (254 nm, 2.5 mW / cm ² ) were irradiated for 1, 5 and 10 minutes, and it was confirmed by gel permeation chromatography (GPC) that there was no change in molecular weight. This result indicates that the axial polymer chain not containing photodegradable nitrobenzene did not respond to ultraviolet irradiation, and α-CD maintained the inclusion state.

[Comparative Synthesis Example 2]
<Synthesis of polyrotaxane X having methacryloyl group as hydrophobic group and polymerizable group (preparation of polyrotaxane Y)>
(9) Addition of polymerizability and hydrophobization of polyrotaxane X having no degradable group (Preparation of polyrotaxane Y)
1 g of polyrotaxane X synthesized in (8) above was dissolved in 15 mL of dehydrated dimethyl sulfoxide, and 209.9 μL of 2-isocyanatoethyl methacrylate and 1.3 mL of butyl isocyanate were added. 1,4-diazabicyclo [2.2.2] octane (500 mg) was added, and the mixture was reacted at room temperature for 24 hours. After the reaction, dialysis was carried out for 2 days against dimethyl sulfoxide using a dialysis membrane (fraction molecular weight 3,500). Subsequently, dialysis was performed for 2 days against water using the same dialysis membrane (fraction molecular weight 3,500).

Example 2 Effect of Reducing Maximum Tensile Strength on Photoinduced Degradation of Cured Product Prepared Using Polyrotaxane B Having Hydrophobic and Polymerizable Groups 29.5 parts by mass of polyrotaxane B synthesized in Synthesis Example 2, 2-hydroxy 69.5 parts by mass of ethyl methacrylate, 0.3 part by mass of 2-dimethylaminoethyl methacrylate, and 0.7 parts by mass of camphorquinone were mixed and used as a solution-type photopolymerizable adhesive B. Photopolymerizable adhesive B was added to a dumbbell mold made of silicone, and light was irradiated for 180 seconds with a dental visible light irradiator (700 mW / cm ² ) to prepare a cured body. The cured product is obtained by adding a photopolymerizable adhesive B to a silicone mold (thickness 1 mm, length 15 mm, center width 1 mm, end width 2 mm) and then applying a visible light irradiator (wavelength (400 to 450 nm, 700 mW / cm ² ) for 180 minutes to produce a cured product. It was confirmed that the nitrobenzyl group of polyrotaxane B was not cleaved under the present light irradiation conditions at wavelengths of 400 to 450 nm. One of the cured bodies was left as it was under shading (non-UV irradiation group, n = 4). One alternative of the cured product is UV irradiation (254nm, 2.5mW / cm ²⁾ for 2 minutes, was allowed to stand under shading (UV irradiated group, n = 4).

In order to evaluate the decomposability of the photopolymerizable adhesive B, a comparative cured product was prepared using the same method as described above. 29.5 parts by mass of polyrotaxane Y synthesized in Comparative Synthesis Example 2, 69.5 parts by mass of 2-hydroxyethyl methacrylate, 0.3 parts by mass of 2-dimethylaminoethyl methacrylate, 0.7 parts by mass of camphorquinone These were mixed and used as a solution-type photopolymerizable adhesive Y. The photopolymerizable adhesive Y was added to a silicone dumbbell mold, and light was irradiated for 180 seconds with a dental visible light irradiator (700 mW / cm ² ) to prepare a cured body. The cured product was obtained by adding a photopolymerizable adhesive Y to a silicone mold (dumb shape having a thickness of 1 mm, a length of 15 mm, a center width of 1 mm, and an end width of 2 mm), and then a visible light irradiator (wavelength). (400 to 450 nm, 700 mW / cm ² ) for 180 minutes to produce a cured product. It was confirmed that the nitrobenzyl group of polyrotaxane Y was not cleaved under the present light irradiation conditions at wavelengths of 400 to 450 nm. One of the cured bodies was left as it was under shading (non-UV irradiation group, n = 4). One alternative of the cured product is UV irradiation (254nm, 2.5mW / cm ²⁾ for 2 minutes, was allowed to stand under shading (UV irradiated group, n = 4). In addition, as a cured product not containing polyrotaxane, a cured product was prepared using Bis-GMA as a crosslinking agent. 29.5 parts by mass of Bis-GMA, 69.5 parts by mass of 2-hydroxyethyl methacrylate, 0.3 parts by mass of 2-dimethylaminoethyl methacrylate, and 0.7 parts by mass of camphorquinone were mixed. Cured in the same manner as above. Tensile tests were conducted using these prepared cured bodies, and the micro tensile strength was calculated from the obtained stress-strain curve. The crosshead speed was measured at 1 mm / min (manufactured by Shimadzu Corp., Autograph EZ Test).

In the Bis-GMA-containing cured product, stress and strain increased linearly in the elastic region, and fractured at about 60 MPa (FIG. 4). However, in the cured product containing polyrotaxane B, the stress hardly changed with respect to the small deformation, and the stress had a linear correlation with the large deformation. In the non-UV irradiation group, the material stretched in the plastic region and then fractured. However, in the UV irradiation group, the cured body was not deformed in the plastic region, and fractured. Further, a large difference was observed in the micro tensile strength before and after UV irradiation in the photopolymerizable adhesive B and the photopolymerizable adhesive Y. When the photopolymerizable adhesive B was used, the micro tensile strength of the non-UV irradiation group was 45.7 MPa (standard deviation 2.4 MPa), whereas the micro tensile strength of the UV irradiation group was 24.1 MPa. The standard deviation was as low as 4.7 MPa (FIG. 5). In the UV irradiation group, it was considered that the microtensile strength was significantly reduced by the decomposition of polyrotaxane B accompanying the cleavage of the photocleavable group (nitrobenzyl group) by UV irradiation. In particular, the micro tensile strength could be greatly reduced by a simple treatment of UV irradiation for a relatively short time. On the other hand, when the photopolymerizable adhesive Y was used, the micro tensile strength of the non-UV irradiation group was 41.3 MPa (standard deviation 4.6 MPa), whereas the micro tensile strength of the UV irradiation group was 38. .2 MPa (standard deviation 4.9 MPa). That is, in the photopolymerizable adhesive B prepared using a polyrotaxane B having a nitrobenzyl group that is a photocleavable group as a decomposable group and a methacryloyl group as a polymerizable group, It was confirmed that the effect of greatly reducing the micro tensile strength of the prepared cured product was obtained by applying a specific wavelength UV light that acts to cleave a certain nitrobenzyl. For example, it is shown that the adhesiveness can be greatly lowered even in a short operation time by performing a simple operation of irradiating the cured product of the photopolymerizable adhesive B with light that acts to break the degradable bond of polyrotaxane B. It was done.

Example 3: Maximum Tensile Strength Reduction Effect on Photo-Induced Decomposition of Cured Product Made by Changing Content of Polyrotaxane B The mass part of polyrotaxane B synthesized in Synthesis Example 2 and polyrotaxane Y synthesized in Comparative Synthesis Example 2 is 9. 5, 29.5, 49.5, 0.3 parts by mass of 2-dimethylaminoethyl methacrylate and 0.7 parts by mass of camphorquinone were fixed, and the total of parts by mass of 2-hydroxyethyl methacrylate was 100 parts by mass. It mixed so that it might become, and was used as a photopolymerization type adhesive agent (FIG. 6). Example 2 is a case where the mass part of the polyrotaxane Y is 29.5, and is described together with Example 3. In the same manner as in Example 2, a photopolymerization adhesive was added to a silicone mold (dumb shape having a thickness of 1 mm, a length of 15 mm, a center width of 1 mm, and an end width of 2 mm), and then a visible light irradiator ( A cured product was produced by irradiation with light at a wavelength of 400 to 450 nm, 700 mW / cm ² ) for 180 minutes. It was confirmed that the nitrobenzyl group of polyrotaxane B was not cleaved under the present light irradiation conditions at wavelengths of 400 to 450 nm. One of the cured bodies was left as it was under shading (non-UV irradiation group, n = 4). Another one of the cured bodies was subjected to UV irradiation (254 nm, 2.5 mW / cm ² ) for 2 minutes and allowed to stand under light shielding (UV irradiation group, n = 4). In the case of the polymerizable adhesive Y that is not UV-decomposable, there was no difference in the microtensile strength between the non-UV irradiation group and the UV irradiation group at any mass part of the polyrotaxane Y (Table 1 and FIG. 7). On the other hand, in the case of the UV-decomposable polymerization adhesive B, when the mass parts are 29.5 and 49.5, the micro tensile strength of the UV irradiation group is 47% and 20%, respectively, compared to the non-UV irradiation group. (Fig. 7), the photopolymerizable adhesive B prepared using polyrotaxane B having a nitrobenzyl group which is a UV-cleavable group has a small amount of cured product prepared by applying UV light. It was confirmed that the effect of greatly reducing the tensile strength was obtained. It is shown that the adhesiveness can be greatly reduced even in a short working time by a simple operation of irradiating the cured product of the photopolymerizable adhesive B with light that acts to break the degradable bond of polyrotaxane B. It was.

The centrally decomposable functional polyrotaxane according to the present invention can be used in various applications by replacing a conventional polyrotaxane having degradable linkers at both ends. Such centrally-resolved functional polyrotaxane has fewer synthetic steps than both-end-resolved type, so there are advantages in manufacturing, and it is also possible to use functional molecules such as peptides that are difficult to introduce at both ends. It becomes. Possible uses include use as a resin monomer in dental materials and use as a therapeutic agent for Niemann-Pick disease type C (NPC), as described above.

Claims

A polyrotaxane comprising a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymer comprises at least two linear polymer moieties linked via at least one degradable moiety Compound.
The linear polymer is represented by the following formula:
X 1 -Y 0 (-Z i -Y i ) i -X 2
Wherein X 1 and X 2 are the same or different end groups, Y 0 and Y i are the same or different linear polymer moieties, and Z i is a degradable moiety , (-Z i -Y i ) i indicates that there are i repeating units composed of a degradable portion Y and a linear polymer portion Z, and each Z i and Y i in the linear polymer is the same The polyrotaxane compound according to claim 1, wherein i is an integer of 1 to 500.
The linear polymer is represented by the following formula:
X 1 -Y 0 -Z 1 -Y 1 -X 2
Wherein X 1 and X 2 are the same or different end groups, Y 0 and Y 1 are the same or different linear polymer moieties, and Z 1 is a degradable moiety. The polyrotaxane compound according to claim 1.
The polyrotaxane compound according to any one of claims 1 to 3, wherein the ratio of the lengths of the two linear polymer portions linked to the degradable portion is contained in 1: 1 to 1: 4.
The polyrotaxane compound according to any one of claims 1 to 4, wherein the degradable moiety is acid decomposable, enzymatic degradable, thermally degradable, or photodegradable.
The degradable moiety includes at least one degradable group, and the degradable group includes a p-methoxyphenacyl group, a 2-nitrobenzyl group, a 2-nitrobenzyloxycarbonyl group, a 2-nitrophenylethylene glycol group, Benzyloxycarbonyl group, 3,5-dimethoxybenzyloxycarbonyl group, α, α-dimethyl-3,5-dimethoxybenzyloxycarbonyl group, 3-nitrophenyl group, 3-nitrophenoxy group, 3,5-dinitrophenoxy group 3-nitrophenoxycarbonyl group, phenacyl group, 4-methoxyphenacyl group, α-methylphenacyl group, 3,5-dimethoxybenzoinyl group, 2,4-dinitrobenzenesulfenyl group, (coumarin-4- Yl) methyl group, 7-nitroindolinyl group, arylazophosphate ester, esthetic Any one of claims 1 to 5, selected from the group consisting of a bond, a Schiff base bond, a carbamate bond, a peptide bond, an ether bond, an acetal bond, a hemiacetal bond, a disulfide bond, an organic peroxide, and an acyl hydrazine bond. The polyrotaxane compound according to one item.
The polyrotaxane compound according to any one of claims 1 to 6, wherein the cyclic molecule is selected from the group consisting of α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin.
The polyrotaxane compound according to any one of claims 1 to 7, wherein the cyclic molecule has one or more substituents.
Substituents are 2-hydroxyethoxyethyl (HEE), hydroxyethyl, hydroxypropyl, hydroxybutyl, hydroxyethoxyethyl, N, N-dimethylaminoethyl, carboxyl, methyl, sulfo, primary Amino group, polyethylene glycol, collagen, transferrin, RGD peptide, oligoarginine, (meth) acryloyl group, (meth) acryloyloxy group, (meth) acryloylamino group, (meth) acryloylthio group, vinyl group, aryl group, styryl The polyrotaxane compound according to claim 8, which is selected from the group consisting of a group and a (meth) acrylamide group.
The linear polymer portion is selected from the group consisting of polyethylene glycol, polypropylene glycol, a copolymer of polyethylene glycol and polypropylene glycol, polyethyleneimine, polyamino acid, and polymethyl vinyl ether. The polyrotaxane compound according to Item.
Terminal groups are 2,4-dinitrophenyl group, 3,5-dinitrophenyl group, cyclodextrin, adamantane group, O-triphenylmethyl (O-Trt) group, S-triphenylmethyl (S-Trt) group, N-triphenylmethyl (N-Trt) group, N-tritylglycine, benzyloxycarbonyl (Z) group, 9-fluorenylmethyloxycarbonyl (Fmoc) group, benzyl ester (OBz) group, tert-butylcarbonyl ( Boc) group, amino acid tertiary butyl ester (OBu group), trityl group, fluorescein, pyrene, substituted benzene, optionally substituted polynuclear aromatic, MPC (2-methacryloyloxyethyl phosphorylcholine), BMA (n-butyl methacrylate) ), A combination of MPC and BMA, and a steroid It is selected from the group, the polyrotaxane compound of any one of claims 1 to 10.
A polyrotaxane compound comprising α-cyclodextrin and a linear molecule having the following structural formula,

Here, n is an integer indicating the number of repeating units of polyethylene glycol,
A polyrotaxane compound in which the linear molecule is polyethylene glycol, the number of α-cyclodextrins is 20 to 40, the terminal group is an adamantane group, and the decomposable group is a 2-nitrobenzyl group.
A polyrotaxane compound comprising α-cyclodextrin having a methacryloyl group and a butyl isocyanate group bonded to each other and a linear molecule having the following structural formula:

Here, n is an integer indicating the number of repeating units of polyethylene glycol,
It has 1 to 10 methacryloyl groups and 5 to 20 butyl isocyanate groups per molecule of α-cyclodextrin, the linear molecule is polyethylene glycol, and the number of α-cyclodextrins is 20 to 40 , A polyrotaxane compound wherein the terminal group is an adamantane group and the decomposable group is a 2-nitrobenzyl group.
A polyrotaxane comprising a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymer comprises at least two linear polymer moieties linked via at least one degradable moiety An adhesive composition containing a compound.
A polyrotaxane comprising a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymer comprises at least two linear polymer moieties linked via at least one degradable moiety A dental material composition comprising a compound.
A polyrotaxane comprising a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymer comprises at least two linear polymer moieties linked via at least one degradable moiety A surface coating agent containing a compound.
A polyrotaxane comprising a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymer comprises at least two linear polymer moieties linked via at least one degradable moiety An anti-adhesion agent comprising a compound.
A polyrotaxane comprising a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymer comprises at least two linear polymer moieties linked via at least one degradable moiety An implant that contains a compound.
A polyrotaxane comprising a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymer comprises at least two linear polymer moieties linked via at least one degradable moiety A tissue regeneration device containing a compound.
A polyrotaxane comprising a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymer comprises at least two linear polymer moieties linked via at least one degradable moiety A pharmaceutical composition comprising a compound for use in the treatment or prevention of a disease.
An adhesive composition, a dental material composition, a surface coating agent, an anti-adhesion agent, an implant, a tissue regeneration device, or a treatment or prevention of a disease comprising the polyrotaxane compound according to claim 12 or 13. A pharmaceutical composition for use.
A polyrotaxane comprising a plurality of cyclic molecules and one linear polymer having a terminal group, wherein the linear polymer comprises at least two linear polymer moieties linked via at least one degradable moiety A method for producing a compound comprising: a) a step of adding a reactive group to both ends of a linear polymer portion; b) a linear polymer portion having a reactive group added to both ends via a degradable portion. Linking to obtain a linear polymer comprising at least two linear polymer moieties linked via at least one degradable moiety; c) reacting the linear polymer with a cyclic molecule to produce a pseudopolyrotaxane And d) adding end groups to both ends of the pseudopolyrotaxane.
The reactive group added to both ends of the linear polymer portion is selected from the group consisting of an amino group, a carboxyl group, an aldehyde group, a sulfanyl group, an azide group, an alkynyl group, a tosyl group, and an active ester group. Item 23. The production method according to Item 22.
14. The method for producing a polyrotaxane compound according to claim 12 or 13, wherein a) a step of adding reactive groups to both ends of the linear polymer portion, b) a line having reactive groups added to both ends. Connecting linear polymer parts via a degradable part to obtain a linear polymer comprising at least two linear polymer parts connected via at least one degradable part, c) linear height A process comprising: reacting a molecule with a cyclic molecule to obtain a pseudopolyrotaxane; and d) adding end groups to both ends of the pseudopolyrotaxane.
The reactive group added to both ends of the linear polymer portion is selected from the group consisting of an amino group, a carboxyl group, an aldehyde group, a sulfanyl group, an azide group, an alkynyl group, a tosyl group, and an active ester group. Item 25. The production method according to Item 24.