Affinity micellar drug complex

Abstract

The present invention provides a novel copolymer useful in drug delivery technology. The present invention relates to a drug complex obtained by binding a target-specific molecule to a copolymer X comprising structural units represented by formulae (A), (B) and (C). [In the formula, R1, R2 and R3 may be the same or different and each represents a hydrogen atom or a C1-3 alkyl group; r4 represents a C1-3 alkyl group; r5 represents a hydrogen atom, a C1-18 alkyl group, a 3-to 8-membered cycloalkyl group which may have a substituent, an adamantyl group, a C6-18 aryl group which may have a substituent, or a 5-to 10-membered heteroaryl group which may have a substituent; x1, X2, and X3 may be the same or different and each represents an oxygen atom, a sulfur atom, or N-R7; r6 represents a hydrogen atom, a leaving group or a linking group; r7 represents a hydrogen atom or a C1-3 alkyl group; m represents an integer of 1 to 100; and n represents an integer of 0-3. ] # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a novel copolymer that can be used in drug delivery technology. More specifically, it relates to a copolymer for use as a drug delivery carrier targeting tumors, a pharmaceutical composition in which the copolymer carries a physiologically active substance such as an anticancer agent, and a pharmaceutical product containing the composition. Background Art

[0002] In recent years, research on drug delivery systems (DDS) has been actively conducted as a technology for efficiently and safely delivering drugs to diseased areas. Among them, as a technology that utilizes the structural characteristics of diseased areas to improve the selectivity of drug accumulation, the demand for DDS using nanoparticles as drug delivery carriers has gradually increased.

[0003] For example, in solid tumor tissue, since the structure of new blood vessels (tumor blood vessels) is not mature compared to normal blood vessels, the intercellular spaces of hundreds of nanometers appear in the vascular endothelium, and the permeability of the substance is high. Using this structural feature, it is known that high molecular weight bodies containing nanoparticles selectively penetrate tumor blood vessels and accumulate in solid tumor tissues. In addition, in solid tumor tissues, the lymphatic system involved in the discharge of macromolecules is dysfunctional, so the infiltrated nanoparticles are continuously retained in the tissue (Enhanced permeability and retention effect, high permeability and retention effect, EPR effect). Conventional low molecular drugs leak out of the blood vessels due to the membrane penetration of vascular cells, so they are non-selectively distributed in the tissues, and are not accumulated in solid tumor tissues. According to the methodology of the EPR effect, in drug delivery using nanoparticles, the distribution in the tissue is dominated by the permeability of the intercellular spaces of the vascular endothelial cells, so the tissue selectivity of solid tumors is improved in the distribution of drugs. Therefore, the EPR effect has become a powerful academic basis for the development of nanotechnology-based pharmaceuticals (nanomedicine) targeting solid tumors.

[0004] It can be considered that the delivery process of medicine in the EPR effect is via blood flow, and the process of nanoparticles leaking out of blood vessels is passively carried out. Therefore, in order to maximize the accumulation of nanoparticles to solid tumors, it is crucial to give the molecular design that can tolerate long-term retention in blood to the constituent components of the nanoparticles that become drug delivery carriers. Therefore, for drug delivery carriers, it is necessary to avoid the ability of obstacles such as non-specific interactions with blood constituents, foreign body recognition realized by the reticuloendothelial system (RES) in liver, spleen and lungs, and glomerular filtration in kidneys. And, it is known that such obstacles can be overcome by particle diameter or by optimizing the particle characteristics such as surface modification utilizing biocompatible polymers. For example, the particle diameter of drug delivery carriers is ideally greater than about 6nm as the kidney clearance threshold and less than 200nm that can avoid utilizing RES identification.

[0005] And, the particle diameter of known drug delivery carriers also can have an impact on the tissue permeability of the affected part.For example, the particle diameter that shows equal blood retention is 30nm, 50nm, 70nm and 100nm medicine inner package nanoparticles tumor suppressor activity is studied comparatively, it is known that the medicine inner package nanoparticles of particle diameter 30nm arrives at the affected part depth, thus shows the best therapeutic effect (non-patent literature 1).Therefore, it can be considered that the particle diameter of the nanoparticles used for the drug delivery carrier targeted with solid tumors is ideally the smallest possible particle diameter in the scope that can avoid kidney clearance.

[0006] Methods of using colloidal dispersions such as liposomes, emulsions or nanoparticles as nanoparticles for drug delivery carriers have been developed; methods of using raw materials of biological origin such as albumin; methods of using natural polymers such as natural polysaccharides; or methods of using synthetic polymers. Among them, synthetic polymers can be used to prepare nanoparticles with precisely controlled particle sizes by appropriately selecting monomers as constituent components and synthesis methods, and therefore are widely used as constituent components of drug delivery carriers.

[0007] For example, the amphiphilic block copolymer composed of a hydrophilic segment and a hydrophobic segment is disclosed as a method for utilizing a drug delivery carrier. The block copolymer is spontaneously associated in an aqueous medium with the hydrophobic interaction between molecules as a driving force to form core-shell nanoparticles (polymeric micelles). The hydrophobic segment of the known polymer micelle can be encapsulated or combined with a low-molecular drug, and the obtained drug encapsulated polymer micelles show higher stability in the blood, and are selectively accumulated in solid tumors by the EPR effect, thus compared to the situation of the solution of the low-molecular drug, it is possible to produce higher anti-cancer activity (patent document 1). However, polymer micelles are an association of multiple molecules, so a particle diameter of about 30nm is a lower limit that can be prepared, and it is very difficult to control the fine size near the particle diameter 10nm that can avoid the influence of renal clearance.

[0008] On the other hand, it is known that in the nanoparticles formed by synthetic polymers, when the driving force forms particles (hereinafter referred to as single-chain nanoparticles (SCNP, single chain nanoparticle)) using chemical crosslinking, hydrophobic interaction, ionic bonds, etc. in the single chain, nanoparticles with a small particle size below 20nm are formed (non-patent literature 2). Therefore, although SCNP is expected to be useful as a drug delivery carrier, a technology that can accurately control its particle size has not yet been developed.

[0009] As another drug delivery technology, there is an antibody-drug complex (ADC) that can deliver cytotoxic agents (drugs) to tumor cells expressing antigens in a targeted manner (non-patent literature 3 to 5). ADC has three components: an antibody (Ab), a linker, and a drug. In order to deliver the drug locally to the target, the drug is linked or conjugated to the antibody. The conjugation method is usually chemically modified by using the amino group of the lysine side chain of the antibody or the cysteine ​​thiol group obtained by reducing the interchain disulfide bond. The design of ADC is still difficult, and many factors must be controlled (such as the choice of antibodies, the stability of the linker, the drug / toxin (payload) and its cleavage dynamics, etc.). Among them, the important parameter is the number of payloads for a single antibody (drug-antibody ratio or DAR (Drug Antibody ratio)). Antibodies that bind a large number of drug / toxin molecules show a binding barrier to the target antigen or a rapid in vivo clearance rate from the bloodstream. Usually, one antibody can only bind a limited number of drug / toxin molecules (usually DAR is 4 to 6). As a result, in order to obtain sufficient effectiveness for killing target cells, it is necessary to use highly toxic agents with an IC50 of less than 1 nM (such as calicheamicin or auristatin monomethyl ester (MMAE)) (Non-patent Documents 6, 7, etc.). Therefore, once a small part of the conjugate is off-target, very large toxic side effects will occur. As mentioned above, a new solution with both high effectiveness and high tolerance is required.

[0010] Affibody molecules, like antibodies, are antibody-mimicking proteins that specifically bind to target antigens (Non-Patent Document 8). They have the characteristics of having specific targeted binding capabilities without the non-specific interactions such as Fc receptor binding exhibited by some antibodies. If used instead of ADC antibodies, drugs can be delivered to targets (e.g., tumor cells). Small engineered scaffold proteins such as affibody molecules are expected to be used as drug carriers.

[0011] The complex of the affinity body molecule and the drug can be a complement or alternative to the current targeted therapy. For example, human epidermal growth factor receptor 2 (HER2) is often overexpressed in various cancers, and the treatment of HER2 targeting is used daily in clinical practice. It is being studied whether the monovalent HER2 binding affinity body molecule (ZHER2: 2891) can be fused with the albumin fusion domain (ABD) and used as a carrier of the cytotoxic emtansine derivative mcDM1. It has been reported that the monovalent HER2 targeted affinity body drug complex has anti-tumor activity in vivo (non-patent document 9). However, the half-life of the affinity body drug complex in the blood is very short. In the case of treatment that uses a high targeting affinity equivalent to that of an antibody to increase tumor accumulation, the absolute amount of uptake in the tumor cannot be obtained, so the dosage needs to be increased.

[0012] Prior art literature

[0013] Patent Literature

[0014] Patent Document 1: Japanese Patent No. 3270592

[0015] Non-patent literature

[0016] Non-patent document 1: H. Cabral et al., Nat. Nanotechnol. 6 815-823 (2011)

[0017] Non-patent literature 2: Jose A. Pomposo, Single-Chain Polymer Nanoparticles: Synthesis, Characterization, Simulations, and Applications (2017)

[0018] Non-patent document 3: Chari, RV et al., Angew. Chem. Int. Ed. 53, 3796-3827 (2014)

[0019] Non-patent document 4: Jagadeesh, D., Smith, MR, Curr. Treat. Options Oncol. 17, 55 (2016)

[0020] Non-patent document 5: Ducry, L., Stump, B., Bioconjugate Chem. 21, 5-13 (2010)

[0021] Non-patent document 6: Casi, G., Neri, D., J. Controlled Release 161, 422-428 (2012)

[0022] Non-patent document 7: Wu, AM, Senter, PD, Nat. Biotechnol. 23, 1137-1146 (2005)

[0023] Non-patent document 8: Nord, K., et al. Nature Biotechnol. 15, 772-777 (1997)

[0024] Non-patent literature 9: Xu, Tianqi et al., Cancers 13(1)85(2020) Summary of the invention

[0025] Problems to be solved by the invention

[0026] The present invention aims to provide a copolymer for a drug delivery carrier targeting a tumor. More specifically, the present invention aims to provide a copolymer for a drug delivery carrier for improving the blood retention and / or tumor accumulation of a drug.

[0027] Technical means to solve problems

[0028] The inventors of the present invention conducted in-depth research to solve the above-mentioned problems, and found that the terpolymer of acrylic acid derivatives has the property of forming SCNP in water. In addition, they successfully created a copolymer for drug delivery carriers that can control SCNP to a micro-scale precision particle size of about 10nm below 20nm and has high tumor accumulation. The polymer was made to carry or bind anticancer agents and administered to subcutaneous transplanted cancer model mice, resulting in excellent anti-tumor effects. Furthermore, a targeted specific micelle drug complex was successfully developed.

[0029] The present invention relates to the following inventions.

[0030] [1] A copolymer comprising a target-specific molecule bonded to a copolymer X having structural units represented by the following formulae (A), (B) and (C).

[0031]

[0032] [Where R 1 , R 2 and R 3 Same or different, indicating hydrogen atoms or C 1-3 Alkyl; R 4 Represents C 1-3 Alkyl; R 5 represents hydrogen atom, C 1-18 alkyl, 3- to 8-membered cycloalkyl which may have a substituent, adamantyl, C 6-18 Aryl or 5- to 10-membered heteroaryl which may have a substituent; X 1 , X 2 and X 3 The same or different, representing oxygen atoms, sulfur atoms or NR 7 ; R 6 represents a hydrogen atom, a leaving group or a connecting group; R 7 Represents a hydrogen atom or C 1-3 Alkyl; m represents an integer of 1 to 100; n represents an integer of 0 to 3.]

[0033] [2] The copolymer according to [1] above, wherein the polymer X is a copolymer formed by polymerizing three monomers represented by the following general formulas (1) to (3).

[0034]

[0035] [Where R 1 , R 2 and R 3 Same or different, indicating hydrogen atoms or C 1-3 Alkyl; R 4 Represents C 1-3 Alkyl; R 5 represents hydrogen atom, C 1-18 alkyl, 3- to 8-membered cycloalkyl which may have a substituent, adamantyl, C 6-18 Aryl or 5- to 10-membered heteroaryl which may have a substituent; X 1 , X 2 and X 3 The same or different, representing oxygen atoms, sulfur atoms or NR 7 ; R 6 represents a hydrogen atom, a leaving group or a connecting group; R 7 Represents a hydrogen atom or C 1-3 Alkyl; m represents an integer of 1 to 100; n represents an integer of 0 to 3.]

[0036] [3] The copolymer according to [1] or [2], wherein R 1 A hydrogen atom.

[0037] [4] The copolymer according to any one of [1] to [3] above, wherein R 2 A hydrogen atom.

[0038] [5] The copolymer described in any one of [1] to [4] above, wherein R 3 A hydrogen atom.

[0039] [6] The copolymer according to any one of [1] to [5], wherein R 4 It is methyl.

[0040] [7] The copolymer according to any one of [1] to [6], wherein R 5 is a C which may have a substituent 6-18 Aryl.

[0041] [8] The copolymer described in any one of [1] to [7] above, wherein R 5 It is phenyl.

[0042] [9] The copolymer described in any one of [1] to [8] above, wherein R 6 A hydrogen atom.

[0043]

[10] The copolymer described in any one of [1] to [8] above, wherein R 6The leaving group is a group represented by the following formula (4).

[0044]

[0045]

[11] The copolymer described in any one of [1] to [8] above, wherein R 6 The linking group is a group represented by the following formula (5).

[0046]

[0047] [Where R 8 Represents a hydrogen atom or a drug; Ak 1 Represents C 1-7 Alkylene bond; X 4 represents an oxygen atom, a sulfur atom or -N(R 7 )-(R 7 Represents a hydrogen atom or C 1-3 alkyl).]

[0048]

[12] The copolymer according to any one of [1] to

[11] , wherein X 1 For oxygen atoms.

[0049]

[13] The copolymer according to any one of [1] to

[12] , wherein X 2 For oxygen atoms.

[0050]

[14] The copolymer according to any one of [1] to

[13] , wherein X 3 is an oxygen atom or NH.

[0051]

[15] The copolymer according to any one of [1] to

[14] above, wherein m is an integer of 4 to 22.

[0052]

[16] The copolymer according to any one of [1] to

[15] above, wherein n is 1.

[0053]

[17] The copolymer described in [1] to

[16] above, wherein the ratio of the structural units (A), (B) and (C) is 0.01 to 100 parts by mass of (B) and 0.1 to 100 parts by mass of (C) per 1 part by mass of (A).

[0054]

[18] The copolymer according to any one of [2] to

[16] above, wherein 0.01 to 100 parts by mass of the monomer (2) and 0.1 to 100 parts by mass of the monomer (3) are polymerized per 1 part by mass of the monomer (1).

[0055]

[19] The copolymer according to any one of [1] to

[18] above, which has a number average molecular weight of 5,000 to 150,000.

[0056]

[20] The copolymer according to any one of [1] to

[19] above, wherein the target-specific molecule is an affinity body.

[0057]

[21] A copolymer as described in any one of [1] to

[19] above, wherein the above-mentioned affinity body is an anti-HER2 affinity body, an anti-HER3 affinity body, an anti-EGFR affinity body, an anti-PD-L1 affinity body, an anti-TNFα affinity body, an anti-fibrinogen affinity body, an anti-transferrin affinity body, an anti-HSA affinity body, an anti-insulin affinity body, an anti-IgG affinity body, an anti-IgM affinity body, an anti-IgA affinity body or an anti-IgE affinity body.

[0058]

[22] The copolymer according to any one of [1] to

[19] above, wherein the affinity body is a molecule comprising an amino acid sequence selected from the group consisting of amino acid sequences (2) to (8).

[0059] AEAKYAKEMRNAYWEIALLPNLTNQQKRAFIRKLYDDPSQSSELLSEAKKLNDSQAPK(sequence number 2)(2)

[0060] AEAKYAKEKYNAYYEIWQLPNLTKYQKAAFIGKLQDDPSQSSELLSEAKKLNDSQAPK(Serial No. 3)(3)

[0061] VDNKFNKEMWAAWEEIRNLPNLNGWQMTAFIASLVDDPSQSANLLAEAKKLNDAQAPK(sequence number 4)(4)

[0062] VDNKFNKEMWIAWEEIRDLPNLNGWQMTAFIASLLDDPSQSANLLAEAKKLNDAQAPK(sequence number 5)(5)

[0063] AEAKYAKEMWIAWEEIRNLPNLNGWQMTAFIAKLLDDPSQSSELLSEAKKLNDSQAPK(sequence number 6)(6)

[0064] AEAKYAKERNAAAYEILYLPNLTNAQKWAFIWKLDDDPSQSSELLSEAKKLNDSQAPK(sequence number 7)(7)

[0065] AEAKYAKERNKAAYEILYLPNLTNAQKWAFIWKLDDDPSQSSELLSEAKKLNDSQAPK(Serial No. 8)(8)

[0066]

[23] A drug complex comprising the copolymer according to any one of [1] to

[22] above and a drug.

[0067]

[24] The drug complex as described in

[23] above, wherein the drug is an antimetabolite, an alkylating agent, an anthracycline, an antibiotic, a mitotic inhibitor, a topoisomerase inhibitor, a proteasome inhibitor or an anti-hormoneagent.

[0068]

[25] The drug complex as described in

[23] above, wherein the drug is DMO, DM1, DM2, DM3, DM4, emtansine, auristatin E, auristatin phenylalanine phenylenediamine (AFP), monomethyl auristatin E, monomethyl auristatin D, monomethyl auristatin F, paclitaxel, docetaxel, irinotecan, topotecan, nogitecan, amsacrine, etoposide, teniposide, mizantrone, SN-38, exatecan or deruxtecan.

[0069]

[26] The drug complex as described in any one of

[23] to

[25] above, wherein the binding between the target specific molecule or drug and the copolymer X is covalent binding or non-covalent binding.

[0070]

[27] A single-chain nanoparticle comprising the copolymer or drug complex described in any one of [1] to

[26] .

[0071]

[28] A pharmaceutical composition comprising the copolymer or drug complex described in any one of [1] to

[27] .

[0072] Effects of the Invention

[0073] According to the examples below, the substance obtained by carrying or combining an anticancer agent with the SCNP obtained by self-association of the copolymer of the present invention shows a tumor growth inhibition effect in a mouse cancer-bearing model, and is therefore suitable for use as a therapeutic agent for malignant tumors. The substance obtained by carrying or combining an anticancer agent with the SCNP obtained by self-association of the copolymer of the present invention can obtain a higher DAR than the existing ADC, and can obtain a high tumor growth inhibition effect with a low dosage, so it is possible to provide a malignant tumor therapeutic agent that has both pharmacological action enhancement and side effect inhibition. In addition, the copolymer of the present invention combined with a targeting-specific molecule can be effectively used for drug delivery using molecules that specifically recognize targets. BRIEF DESCRIPTION OF THE DRAWINGS

[0074] Figure 1 The copolymer obtained in Example 1 was measured by nuclear magnetic resonance spectroscopy (NMR). 1 Figure 1 H-NMR spectrum.

[0075] Figure 2 This is a diagram showing a chromatogram of the copolymer obtained in Example 1 measured by gel permeation chromatography (GPC).

[0076] Figure 3 It is a graph showing the particle size measurement results (scattering intensity distribution) of the copolymer before DACHPt encapsulation (Example 69) and DACHPt encapsulating SCNP (Example 70) by dynamic light scattering (DLS).

[0077] Figure 4 The N3-poly[(benzyl acrylate)-(poly(ethylene glycol) methyl ether acrylate)-(acrylic acid) copolymer] obtained in Example 72 was measured using nuclear magnetic resonance spectroscopy (NMR). 1 Figure 1 H-NMR spectrum.

[0078] Figure 5 This is a diagram showing the chromatogram of the N3-poly[(benzyl acrylate)-(poly(ethylene glycol) methyl ether acrylate)-(acrylic acid) copolymer] obtained in Example 72 measured by gel permeation chromatography (GPC).

[0079] Figure 6The 1,4-diaminobutane-bound copolymer obtained in Example 73 was measured using nuclear magnetic resonance spectroscopy (NMR). 1 Figure 1 H-NMR spectrum.

[0080] Figure 7 The DM1-SPDP-1,4-diaminobutane copolymer obtained in Example 74 was measured using nuclear magnetic resonance spectroscopy (NMR). 1 Figure 1 H-NMR spectrum.

[0081] Figure 8 This is a graph showing the particle size measurement results (scattering intensity distribution) of the anti-HER2 affibody micelle drug complex obtained in Example 75 using the dynamic light scattering (DLS) method.

[0082] Fig. 9 This is a graph showing the particle size measurement results (scattering intensity distribution) of the anti-EGFR affibody micelle drug complex obtained in Example 76 by dynamic light scattering (DLS).

[0083] Fig.10 This is a graph showing the particle size measurement results (scattering intensity distribution) of the anti-HER3 affibody micelle drug complex obtained in Example 77 using the dynamic light scattering (DLS) method.

[0084] Fig.11 This is a graph showing the change in relative tumor volume when an oxaliplatin solution or DACHPt-encapsulated SCNP (Example 70) was administered three times every other day to a mouse colon cancer cell line (C26) subcutaneously transplanted model mouse on the back. DETAILED DESCRIPTION

[0085] The terms used in this specification are used according to their common meanings in the art, except for those otherwise specified.

[0086] In this specification, the term "nanoparticle" refers to a structure having a particle size of 100 nm or less.

[0087] In this specification, "single chain nanoparticles (SCNP)" refer to nanoparticles formed by chemical crosslinking, hydrophobic interaction, ionic bonding, etc. within a single chain as a driving force. In most cases, it means a relatively small particle size of 20 nm or less among nanoparticles.

[0088] In this specification, the "initiator" refers to an initiator for thermal radical polymerization such as an azo compound or a peroxide.

[0089] In the present specification, the "chain transfer agent" refers to a compound that causes a chain transfer reaction in radical polymerization, and is preferably a compound having a thiocarbonyl group.

[0090] In this specification, the term “C 1-3 The term "alkyl" refers to a linear or branched alkyl group having 1 to 3 carbon atoms, and examples thereof include methyl, ethyl, n-propyl and isopropyl.

[0091] In this specification, the term “C 1-18 The term "alkyl" refers to a linear or branched alkyl group having 1 to 18 carbon atoms, and examples thereof include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, and the like.

[0092] In the present specification, the term "3- to 8-membered cycloalkyl group which may have a substituent" means a cyclic alkyl group having 3 to 8 carbon atoms, and examples thereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. The substituent is not particularly limited, and examples thereof include halogen atoms, alkyl groups having 1 to 6 carbon atoms, alkenyl groups having 2 to 6 carbon atoms, alkynyl groups having 2 to 6 carbon atoms, hydroxyl groups, alkoxy groups having 1 to 6 carbon atoms, amino groups, alkylamino groups having 1 to 6 carbon atoms, dialkylamino groups having 1 to 6 carbon atoms in which the alkyl groups are the same or different, thiol groups, alkylthio groups having 1 to 6 carbon atoms, carboxyl groups, alkoxycarbonyl groups having 1 to 6 carbon atoms, and carbamoyl groups.

[0093] In this specification, the term "C 6-18 "Aryl" refers to a monocyclic or condensed-ring polycyclic aromatic hydrocarbon group, for example, phenyl, naphthyl, anthracenyl, phenanthrenyl, terphenyl, pyrene, In addition, the term "C optionally substituted" refers to 6-14The term "aryl" refers to a monocyclic or condensed-ring polycyclic aromatic hydrocarbon group, and examples thereof include phenyl, naphthyl, anthryl, phenanthryl, and the like. The substituent is not particularly limited, and examples thereof include a halogen atom, an alkyl group having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, an alkynyl group having 2 to 6 carbon atoms, a hydroxyl group, an alkoxy group having 1 to 6 carbon atoms, an amino group, an alkylamino group having 1 to 6 carbon atoms, two alkylamino groups having 1 to 6 carbon atoms in which the alkyl groups are the same or different, a thiol group, an alkylthio group having 1 to 6 carbon atoms, a carboxyl group, an alkoxycarbonyl group having 1 to 6 carbon atoms, and a carbamoyl group.

[0094] In the present specification, the term "5- to 10-membered heteroaryl group which may have a substituent" refers to a 5- to 10-membered monocyclic aromatic heterocyclic group or a condensed aromatic heterocyclic group which contains, as ring atoms, 1 to 4 heteroatoms selected from nitrogen atoms, oxygen atoms and sulfur atoms in addition to carbon atoms. Examples of the monocyclic aromatic heterocyclic group include furanyl, thienyl, pyrrolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, isoxazolyl, 1,3,4-thiadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, tetrazolyl and the like. Examples of the condensed aromatic heterocyclic group include benzofuranyl, benzothienyl, quinoxalinyl, indolyl, isoindolyl, isobenzofuranyl, chromanyl group, benzimidazolyl, benzothiazolyl, benzoxazolyl, quinolyl, isoquinolyl, etc. In addition, the so-called "6- to 10-membered heteroaryl group which may have a substituent" refers to a 6- to 10-membered monocyclic aromatic heterocyclic group or a condensed aromatic heterocyclic group which contains 1 to 4 heteroatoms selected from nitrogen atoms, oxygen atoms, and sulfur atoms as ring atoms in addition to carbon atoms. Examples of the monocyclic aromatic heterocyclic group include pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, etc. Examples of the condensed aromatic heterocyclic group include benzofuranyl, benzothienyl, quinoxalinyl, indolyl, isoindolyl, isobenzofuranyl, chromanyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, quinolyl, isoquinolyl, etc. The substituent is not particularly limited, and examples thereof include halogen atoms, alkyl groups having 1 to 6 carbon atoms, alkenyl groups having 2 to 6 carbon atoms, alkynyl groups having 2 to 6 carbon atoms, hydroxyl groups, alkoxy groups having 1 to 6 carbon atoms, amino groups, alkylamino groups having 1 to 6 carbon atoms, two alkylamino groups having 1 to 6 carbon atoms in which the alkyl groups are the same or different, thiol groups, alkylthio groups having 1 to 6 carbon atoms, carboxyl groups, alkoxycarbonyl groups having 1 to 6 carbon atoms, and carbamoyl groups.

[0095] In the present specification, examples of the "halogen atom" include a fluorine atom, a chlorine atom, a bromine atom and an iodine atom.

[0096] In this specification, the term “C 1-7The term "alkylene bond" refers to an optionally substituted linear or branched alkylene group having 1 to 7 carbon atoms, and examples thereof include methylene, ethylene, propylene, butylene, pentylene, hexylene, heptylene, -CH(CH3)-, -C(CH3)2-, -CH(CH2CH3)-, -CH(CH3)CH2-, -CH(CH2CH2CH3)-, -CH(CH2(CH3)2)-, -C(CH3)(CH2CH3)-, -C(CH3)2CH2-, -CH(CH2CH3)CH2-, -CH(CH3)CH(CH3)-, -CH(CH3)CH2CH2-, -CH2CH(CH3)CH2-, -CH(CH2C H2CH2CH3)-, -C(CH3)(CH2CH2CH3)-, -C(CH2CH3)2-, -CH(CH2CH2CH3)CH2-, -CH(CH2(CH3)2)CH2-, -C(CH3)(CH2CH3)CH2-, -C(CH3)2CH(CH3)-, -CH (CH2CH3)CH(CH3)-, -C(CH3)2CH2CH2-, -CH(CH2CH3)CH2CH2-, -CH2CH(CH2CH3)CH2-, -CH(CH3)CH(CH3)CH2-, -CH(CH3)CH2CH(CH3)-, -CH(CH2CH2CH 2CH2CH3)-, -C(CH3)(CH2CH2CH2CH3)-, -C(CH2CH3)(CH2CH2CH3)-, -C(CH2CH3)(CH2(CH3)2)-, -CH(CH2CH2CH2CH3)CH2-, -CH(CH(CH3)CH2CH3)CH2 -, -CH(CH2CH(CH3)CH3)CH2-, -CH(CH2CH2(CH3)2)CH2-, -C(CH3)(CH2CH2CH3)CH2-, -C(CH2CH3)2CH2-, -CH(CH2(CH3)2)CH2-, -C(CH3)(CH2CH3)CH( CH3)-, -CH(CH2CH3)C(CH3)2-, -CH(CH2CH3)CH(CH2CH3)-, -C(CH3)2C(CH3)2-, -CH(CH2CH2CH3)CH2CH2-, -CH(CH2(CH3)2)CH2CH2-, -CH2CH(CH2CH2 CH3)CH2-, -CH2CH(CH2(CH3)2)CH2-, -C(CH3)(CH2CH3)CH2CH2-, -CH(CH2CH3)CH(CH3)CH2-, -CH(CH2CH3)CH2CH(CH3)-, -CH2CH(CH2CH3)CH(CH3)-,-CH(CH3)CH(CH3)CH(CH3)-、-C(CH3)2CH(CH3)CH2-、-C(CH3)2CH2CH(CH3)-、-CH(CH2CH3)CH2CH2CH2-、-CH2CH(CH2CH3)CH2CH2-、-C(CH3)2CH2CH2C H2-、-CH(CH3)CH(CH3)CH2CH2-、-CH(CH3)CH2CH(CH3)CH2-、-CH(CH3)CH2CH2CH(CH3)-、-CH(CH3)CH2CH2CH2CH2-、-CH2CH(CH3)CH2CH2CH2-、-CH2CH2 CH(CH3)CH2CH2-、-C(CH2CH3)(CH2CH2CH2CH3)-、-C(CH2CH2CH3)2-、-C(CH3)(CH2CH2CH2CH3)CH2-、-C(CH3)(CH(CH3)CH2CH3)CH2-、-C(CH3)(CH2CH (CH3)CH3)CH2-、-C(CH3)(CH2CH2(CH3)2)CH2-、-CH(CH2CH2CH2CH3)CH(CH3)-、-CH(CH(CH3)CH2CH3)CH(CH3)-、-CH(CH2CH(CH3)CH3)CH(CH3)-、-CH( CH2CH2(CH3)2)CH(CH3)-、-C(CH3)(CH2CH2CH3)CH(CH3)-、-C(CH2CH3)2CH(CH3)-、-CH(CH2(CH3)2)CH(CH3)-、-C(CH3)(CH2CH3)C(CH3)2-、-C(CH3) (CH2CH2CH3)CH2CH2-、-CH(CH2CH2CH3)CH(CH3)CH2-、-CH(CH2CH2CH3)CH2CH(CH3)-、-CH2CH(CH2CH2CH3)CH(CH3)-、-C(CH3)(CH2CH3)CH(CH3)CH2-、 -C(CH3)(CH2CH3)CH2CH(CH3)-、-CH(CH2CH3)C(CH3)2CH2-、-CH(CH2CH3)CH(CH3)CH(CH3)-、-CH(CH2CH3)CHCH(CH3)2-、-C(CH3)2CH(CH2CH3)CH2-、-CH(CH3)C(CH3)(CH2CH3)CH2-、-CH(CH3)CH(CH2CH3)CH(CH3)-、-C(CH3)2CH(CH3)CH(CH2CH3)CH(CH3)-、-CH(CH3)C(CH3)2CH(CH3)-、-C(CH3)2CH(CH3)CH2CH2-、-C(CH3)2CH2CH(CH3)CH2-, -C(CH3)2CH2CH2CH(CH3)-, -CH(CH3)C(CH3)2CH2CH2-, -CH(CH3)CH(CH3)CH(CH3)CH2-, -CH(CH3)CH(CH3)CH2CH(CH3)-, -CH(CH3)CH(CH3)CH2CH(CH3)-, -CH(CH3)CH(CH3)CH2CH2CH2-, -CH(CH3)CH(CH3)CH2CH2CH2-, -CH(CH3)CH2CH(CH3)CH2CH2-, -CH(CH3)CH2CH2CH(CH3)CH2-, -CH(CH3)CH2CH2CH(CH3)CH2-, -C(CH3)2CH2CH2CH2CH2-, or -CH(CH2CH3)CH2CH2CH2CH2-. The substituent is not particularly limited, and any hydrogen atom may be substituted, and examples thereof include phenoxy, carboxylic acid, sulfonic acid, phosphoric acid, hydroxyl, or thiol groups. ,

[0097] In the present specification, the so-called "targeting specific molecule" is a molecule that recognizes and specifically binds to a marker or receptor (such as a transmembrane protein, a surface insoluble protein or a proteoglycan) expressed on the cell surface, and is a molecule that specifically recognizes a target that is unique to or overexpressed by cancer cells and required for the proliferation or metastasis of cancer, and is a peptide or peptidomimetic drug. Examples of peptides or peptidomimetic drugs include: affibody molecules (such as ABY-025), integrin targeting peptides (RGD peptides), LHRH receptor targeting peptides, ErbB2 (HER2) receptor targeting peptides, prostate-specific membrane-bound antigen (PSMA) targeting peptides, lipoprotein receptor LRP1 targeting peptides, peptides from ApoE protein, ApoA protein peptides, somatostatin receptor targeting peptides, peptides from chlorotoxin, bombesin, insulin, transferrin, fibrinogen-γ fragment, thrombospondin, claudin, ankyrin repeat protein, ankyrin-like repeat protein or synthetic peptides, etc. Hereinafter, the copolymer X of the present invention to which a targeting-specific molecule is bonded or supported may be referred to as a "targeting-specific copolymer".

[0098] In this specification, the so-called "affine" is a polypeptide that can specifically bind to a target antigen. It is the B domain of the IgG binding domain from Staphylococcus protein A (protein A produced by Staphylococcus aureus) and has three α-helical structures. In addition, there is a Z domain as a synthetic IgG binding domain.

[0099] The wild-type sequence thereof comprises the 58 amino acid sequence shown in the following amino acid sequence number (1).

[0100] VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSAN LLAEAKKLNDAQAPK(1)

[0101] In order to improve the specific affinity with the target, the affinity molecule adopts a sequence in which one or more amino acids are deleted, substituted, inserted or added synchronously or asynchronously. Preferably, positions 9, 10, 11, 13, 14, 17, 18, 24, 25, 27, 28, 32 and 35 are substituted with any amino acid. Molecules that specifically bind to target antigens (e.g., HER-2, EGFR) can be selected from a dedicated affinity library (e.g., U.S. Patent No. 5,831,012) according to screening methods known in the art. Affibody molecules can be monomers, or one or more affinity molecules of the same or different targeting specificities can be used in combination. As different targets, for example, albumin binding affinity bodies described in International Publication No. 2009 / 016043 can be cited. In addition, it can also be used in conjunction with a weak electrophilic agent covalently bonded to the target.

[0102] Characteristics of Affibody molecules include high solubility, tissue penetration, stability to heat and enzymes, and relatively low production cost.

[0103] The affibody molecule can be supported on the copolymer X of the present invention through electrostatic interaction, hydrogen bonding, hydrophobic interaction, covalent bonding, etc. It can be modified within the range of maintaining binding specificity and can be bound to the copolymer X via the head end (position 1) or the tail end (position 58).

[0104] In this specification, the so-called "pharmaceutical composition" refers to a substance in which an active ingredient (drug, physiologically active substance) that can be used for diagnosis, prevention or treatment of a disease is supported on a target-specific copolymer or drug complex in which a target-specific molecule is bound to the copolymer X of the present invention through electrostatic interaction, hydrogen bonding, hydrophobic interaction or covalent bonding. As the supported form, when the target-specific copolymer or drug complex forms nanoparticles, examples include a form in which the drug is present on the surface of the particle, a form in which the drug is enclosed in the nanoparticle, or a form in which the drug is combined.

[0105] One embodiment of the present invention is a copolymer or drug complex in which a target-specific molecule is bonded to a copolymer X having structural units represented by the following formulae (A), (B) and (C).

[0106]

[0107] [Where R 1 , R 2and R 3 Same or different, indicating hydrogen atoms or C 1-3 Alkyl; R 4 Represents C 1-3 Alkyl; R 5 represents hydrogen atom, C 1-18 alkyl, 3- to 8-membered cycloalkyl which may have a substituent, adamantyl, C 6-18 Aryl or 5- to 10-membered heteroaryl which may have a substituent; X 1 , X 2 and X 3 The same or different, representing oxygen atoms, sulfur atoms or NR 7 ; R 6 represents a hydrogen atom, a leaving group or a connecting group; R 7 Represents a hydrogen atom or C 1-3 Alkyl; m represents an integer of 1 to 100; n represents an integer of 0 to 3.]

[0108] In the copolymer X of the present invention, the structural unit (A) functions as a unit that imparts hydrophilicity, and the structural unit (B) functions as a unit that imparts hydrophobicity. In addition, the structural unit (C) functions as a scaffold for binding an active ingredient (drug, physiologically active substance) to the copolymer X or the targeting-specific copolymer. The copolymer X, the targeting-specific copolymer or the drug complex of the present invention has these three structural units, and therefore has the property of forming SCNPs in water, and can achieve precise particle size control of the formed SCNPs to a microscale of less than 20 nm, and can function as a drug delivery carrier with high tumor accumulation.

[0109] R in the structural unit (A) 1 Represents a hydrogen atom or C 1-3 The alkyl group is preferably a hydrogen atom or a methyl group, further preferably a hydrogen atom, an ethyl group or a propyl group, and more preferably a hydrogen atom.

[0110] X 1 represents an oxygen atom, a sulfur atom or NR 7 , preferably an oxygen atom, a sulfur atom or NH, more preferably an oxygen atom.

[0111] m represents an integer of 1 to 100, preferably an integer of 3 to 100, and is preferably 3 to 80, more preferably 4 to 60, further preferably 4 to 40, and further preferably 4 to 22 from the viewpoint of imparting good hydrophilicity.

[0112] R 4 Represents C 1-3 The alkyl group is specifically a methyl group, an ethyl group, an n-propyl group or an isopropyl group, preferably a methyl group or an ethyl group, and more preferably a methyl group.

[0113] R in the structural unit (B) 2 Represents a hydrogen atom or C 1-3 The alkyl group is preferably a hydrogen atom or a methyl group, further preferably a hydrogen atom, an ethyl group or a propyl group, and more preferably a hydrogen atom.

[0114] X 2 represents an oxygen atom, a sulfur atom or NR 7 , preferably an oxygen atom, a sulfur atom or NH, more preferably an oxygen atom.

[0115] n represents an integer of 0 to 3, preferably an integer of 1 to 3, and more preferably 1.

[0116] R 5 represents hydrogen atom, C 1-18 alkyl, 3- to 8-membered cycloalkyl which may have a substituent, adamantyl, C 6-18 The aryl group or the 5- to 10-membered heteroaryl group which may have a substituent is preferably C 1-18 alkyl, 3- to 8-membered cycloalkyl which may have a substituent, adamantyl, C 6-18 Aryl or 5- to 10-membered heteroaryl which may have a substituent, more preferably C 1-18 alkyl, 3- to 8-membered cycloalkyl which may have a substituent, adamantyl, C 6-18 An aryl group or an optionally substituted 5- to 10-membered heteroaryl group, more preferably a C 1-18 Alkyl, 3-8 membered cycloalkyl, adamantyl or C 6-18 On the other hand, 3- to 8-membered cycloalkyl group which may have a substituent, adamantyl group, C 6-14 Aryl or 6- to 10-membered heteroaryl group which may have a substituent. Here, the substituent is preferably one or more selected from the group consisting of a halogen atom, an alkyl group having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, and an alkynyl group having 2 to 6 carbon atoms.

[0117] R in the structural unit (C) 3 Represents a hydrogen atom or C 1-3 The alkyl group is preferably a hydrogen atom or a methyl group, and is also preferably a hydrogen atom, an ethyl group or a propyl group, and is more preferably a hydrogen atom.

[0118] X 3 represents an oxygen atom, a sulfur atom or NR 7 , preferably an oxygen atom, a sulfur atom or NH, more preferably an oxygen atom or NH.

[0119] R 6represents a hydrogen atom, a leaving group or a linking group. The leaving group is a group that can be detached when the structural unit (C) is combined with the drug (physiologically active substance), and the linking group is a group that can be used for cross-linking when the structural unit (C) is combined with the drug (physiologically active substance). As these leaving groups or linking groups, C which may have a substituent is preferred. 1-18 Alkyl, 3- to 8-membered cycloalkyl which may have a substituent, C 7-19 Aralkyl. Here, as the substituent, for example, there can be mentioned a halogen atom, an alkyl group having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, an alkynyl group having 2 to 6 carbon atoms, a hydroxyl group, an alkoxy group having 1 to 6 carbon atoms, an amino group, an alkylamino group having 1 to 6 carbon atoms, two alkylamino groups having 1 to 6 carbon atoms in which the alkyl groups are the same or different, a thiol group, an alkylthio group having 1 to 6 carbon atoms, a carboxyl group, an alkoxycarbonyl group having 1 to 6 carbon atoms, a carbamoyl group, etc. Among these groups, as the linking group, a group having a functional group such as a hydroxyl group, an amino group, a thiol group, a carboxyl group, etc. as a substituent is preferred.

[0120] As R 6 A preferred specific example of the leaving group of may be a group represented by the following formula (4).

[0121]

[0122] As R 6 Preferred specific examples of the linking group include a group represented by the following formula (5).

[0123]

[0124] [Where R 8 Represents a hydrogen atom or a drug; Ak 1 Represents C 1-7 Alkylene bond; X 4 represents an oxygen atom, a sulfur atom or -N(R 7 )-(R 7 Represents a hydrogen atom or C 1-3 alkyl).]

[0125] In formula (5), it is more preferred that X 4 represents an oxygen atom, a sulfur atom or NH.

[0126] R 6 Among the linking groups, more preferably, a group selected from the following formulae (6) to (8) is exemplified.

[0127]

[0128] The copolymer X of the present invention is a copolymer having structural units shown in formulas (A), (B) and (C). The copolymer X may be a random copolymer or a block copolymer, preferably a random copolymer. Regarding the composition ratio of each structural unit in one molecule, it is preferred that when (A) is set to 1 part by mass, (B) is 0.01 to 100 parts by mass and (C) is 0.1 to 100 parts by mass; more preferably, when (A) is set to 1 part by mass, (B) is 0.05 to 18 parts by mass and (C) is 0.1 to 20 parts by mass; particularly preferably, when (A) is set to 1 part by mass, (B) is 0.05 to 4 parts by mass and (C) is 0.1 to 16 parts by mass.

[0129] The degree of polymerization of the copolymer X of the present invention is not particularly limited, but is preferably 5,000 to 150,000, more preferably 8,000 to 150,000 as a number average molecular weight.

[0130] In the copolymer of the present invention, as described above, the monomer represented by the general formula (1) functions as a unit imparting hydrophilicity, and the monomer represented by the general formula (2) functions as a unit imparting hydrophobicity. In addition, the monomer represented by the general formula (3) functions as a scaffold for combining drugs with the copolymer. As the monomer represented by the general formula (2) that functions as a hydrophobic unit, for example, the monomer represented by the following formula can be exemplified.

[0131]

[0132]

[0133] In the general formula (1), R 1 Represents a hydrogen atom or C 1-3 The alkyl group is preferably a hydrogen atom or a methyl group, further preferably a hydrogen atom, an ethyl group or a propyl group, and more preferably a hydrogen atom.

[0134] In the general formula (2), R 2 Represents a hydrogen atom or C 1-3 The alkyl group is preferably a hydrogen atom or a methyl group, further preferably a hydrogen atom, an ethyl group or a propyl group, and more preferably a hydrogen atom.

[0135] In the general formula (3), R 3 Represents a hydrogen atom or C 1-3 The alkyl group is preferably a hydrogen atom or a methyl group, further preferably a hydrogen atom, an ethyl group or a propyl group, and more preferably a hydrogen atom.

[0136] In the general formula (1), R 4 Represents C 1-3 The alkyl group is specifically a methyl group, an ethyl group, an n-propyl group or an isopropyl group, preferably a methyl group or an ethyl group, and more preferably a methyl group.

[0137] In the general formula (1), X 1represents an oxygen atom, a sulfur atom or NR 7 , preferably an oxygen atom, a sulfur atom or NH, more preferably an oxygen atom.

[0138] In the general formula (1), m represents an integer of 1 to 100, preferably an integer of 3 to 100, and is preferably 3 to 80, more preferably 4 to 60, further preferably 4 to 40, and further preferably 4 to 22 from the viewpoint of imparting good hydrophilicity.

[0139] In the general formula (2), R 5 represents hydrogen atom, C 1-18 alkyl, 3- to 8-membered cycloalkyl which may have a substituent, adamantyl, C 6-18 The aryl group or the 5- to 10-membered heteroaryl group which may have a substituent is preferably C 1-18 alkyl, 3- to 8-membered cycloalkyl which may have a substituent, adamantyl, C 6-18 Aryl or 5- to 10-membered heteroaryl which may have a substituent, more preferably C 1-18 alkyl, 3- to 8-membered cycloalkyl which may have a substituent, adamantyl, C 6-18 An aryl group or an optionally substituted 5- to 10-membered heteroaryl group, more preferably a C 1-18 Alkyl, 3-8 membered cycloalkyl, adamantyl or C 6-18 On the other hand, 3- to 8-membered cycloalkyl groups which may have a substituent, adamantyl groups, C 6-14 Aryl or 6- to 10-membered heteroaryl group which may have a substituent. Here, the substituent is preferably one or more selected from the group consisting of a halogen atom, an alkyl group having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, and an alkynyl group having 2 to 6 carbon atoms.

[0140] In the general formula (2), X 2 represents an oxygen atom, a sulfur atom or NR 7 , preferably an oxygen atom, a sulfur atom or NH, more preferably an oxygen atom.

[0141] In the general formula (2), n represents an integer of 0 to 3, preferably an integer of 1 to 3, and more preferably 1.

[0142] In the general formula (3), R 6 represents a hydrogen atom, a leaving group or a linking group. As these leaving groups or linking groups, preferably C 1-18 Alkyl, 3- to 8-membered cycloalkyl which may have a substituent, C 7-19Aralkyl. Here, as the substituent, for example, there can be mentioned a halogen atom, an alkyl group having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, an alkynyl group having 2 to 6 carbon atoms, a hydroxyl group, an alkoxy group having 1 to 6 carbon atoms, an amino group, an alkylamino group having 1 to 6 carbon atoms, two alkylamino groups having 1 to 6 carbon atoms in which the alkyl groups are the same or different, a thiol group, an alkylthio group having 1 to 6 carbon atoms, a carboxyl group, an alkoxycarbonyl group having 1 to 6 carbon atoms, a carbamoyl group, etc. Among these groups, as the linking group, a group having a functional group such as a hydroxyl group, an amino group, a thiol group, a carboxyl group, etc. as a substituent is preferred.

[0143] As R 6 Preferred specific examples of the leaving group include a group represented by the following formula (4).

[0144]

[0145] As R 6 As a specific example of a preferred linking group, a group represented by the following formula (5) is preferred.

[0146]

[0147] [Where R 8 Represents a hydrogen atom or a drug; Ak 1 Represents C 1-7 Alkylene bond; X 4 represents an oxygen atom, a sulfur atom or -N(R 7 )-(R 7 Represents a hydrogen atom or C 1-3 alkyl).]

[0148] In formula (5), it is more preferred that X 4 It represents an oxygen atom, a sulfur atom or an NH group.

[0149] R 6 Among the linking groups, more preferably, a group selected from the following formulae (6) to (8) is exemplified.

[0150]

[0151] In the general formula (3), X 3 represents an oxygen atom, a sulfur atom or NR 7 , preferably an oxygen atom, a sulfur atom or NH, more preferably an oxygen atom or NH.

[0152] The copolymer X of the present invention is formed by copolymerizing three monomers represented by general formulas (1) to (3). The copolymerization may be random copolymerization or block copolymerization, and is preferably formed by random copolymerization. Regarding the three monomer mixing ratios, when the mass part of monomer (1) is set to 1, it is preferred to polymerize 0.01 to 100 mass parts of monomer (2) and 0.1 to 100 mass parts of monomer (3), more preferably 0.05 to 18 mass parts of monomer (2) and 0.1 to 20 mass parts of monomer (3), and particularly preferably 0.05 to 4 mass parts of monomer (2) and 0.1 to 16 mass parts of monomer (3).

[0153] In addition, "solvates" coordinated with various solvents are also included in the copolymer X of the present invention. In the present specification, "solvates" include, for example, hydrates and ethanolates. The solvent may be coordinated with the copolymer X of the present invention in any amount.

[0154] The copolymer X of the present invention can be produced by various known methods. The production method is not particularly limited, and for example, the copolymer can be produced according to the basic polymer synthesis method described below.

[0155]

[0156] [wherein, R' represents a hydrogen atom or C 1-3 Alkyl, R" represents the above R 4 , R 5 or R 6 The groups shown.]

[0157] This reaction represents a step of making a monomer (I) react with a chain transfer agent (II) and an initiator to produce a polymer (III). This reaction can be carried out in the absence of a solvent or in the following solvents: alcohols such as methanol, ethanol, 1-propanol, and 2-propanol; ethers such as diethyl ether, tetrahydrofuran, and 1,4-dioxane; aromatic hydrocarbons such as benzene, toluene, and xylene; halogenated hydrocarbons such as dichloromethane, chloroform, and 1,2-dichloroethane; N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, acetonitrile, ethyl acetate, etc., preferably aromatic hydrocarbons such as toluene and xylene are used as solvents. As the chain transfer agent, 2-(dodecylthiocarbonylthio)-2-methylpropionic acid (DDMAT), cyanomethyl dodecyl trithiocarbonate (CDTTC), 2-cyano-2-propyl dodecyl trithiocarbonate (CPDTTC), 4-cyano-4-[(dodecylthiocarbonylthio)thio]pentanoic acid (CDSPA), 2-(dodecylthiocarbonylthio)-2-methylpropionic acid-3-azido-1-propanol ester (N3-CTA) and the like can be used. Preferably, DDMAT or N3-CTA is used, and more preferably, N3-CTA is used. When the chain transfer agent is used for polymerization, the copolymer X of the present invention forms a structure in which a part or all of the structure of the chain transfer agent is partially bonded. When the copolymer X contains the structure of the chain transfer agent, the structure can be removed by an appropriate method. As an initiator, azobisisobutyronitrile (AIBN), 1,1'-azobis(cyclohexanecarbonitrile) (ACHN), 2,2'-azobis-2-methylbutyronitrile (AMBN), 2,2'-azobis-2,4-dimethylvaleronitrile (ADVN), 2,2'-azobis(dimethyl 2-methylpropionate) (MAIB) and the like azo polymerization initiators can be used, preferably AIBN. The reaction temperature is 0 to 300°C, preferably 0 to 150°C, more preferably 1 to 100°C, and the reaction time is 1 minute to 48 hours, preferably 5 minutes to 24 hours. In this reaction, the reaction can be carried out in the presence of monomers (I) of different structures to produce a random copolymer X.

[0158] For example, when N3-CTA having an azide group is used as a chain transfer agent, a copolymer X having an azide group at the terminal can be obtained, which is advantageous for producing a target-specific copolymer or a drug conjugate by a click reaction described later.

[0159] As the targeting specific molecule bound to the above-mentioned copolymer X, an affinity molecule can be mentioned, for example, an anti-ErbB2 affinity (also known as an anti-HER2 affinity), an anti-HER3 affinity, an anti-EGFR affinity, an anti-PD-L1 affinity, an anti-TNFα affinity, an anti-fibrinogen affinity, an anti-transferrin affinity, an anti-HSA affinity, an anti-insulin affinity, an anti-IgG affinity, an anti-IgM affinity, an anti-IgA affinity or an anti-IgE affinity (for example, available from Abcam, Cambridge, MA), etc., and more preferably an anti-EGFR affinity, an anti-HER2 affinity, an anti-HER3 affinity, and an anti-PD-L1 affinity.

[0160] Preferred affibody molecules include molecules described in International Publication No. 2009 / 080810 (anti-HER2), International Publication No. 2011 / 056124 (anti-HER3), International Publication No. 2014 / 053586 (anti-HER3), International Publication No. 2007 / 065635 (anti-EGFR) or International Publication No. 2017 / 072280 (anti-PD-L1). As modifications within the range of maintaining binding specificity, the affibody molecule can be modified with one or more amino acids.

[0161] A more preferred affibody molecule is an anti-HER2 affibody comprising the sequence shown in the amino acid sequence (2) known as Z (HER2:2891).

[0162] AEAKYAKEMRNAYWEIALLPNLTNQQKRAFIRKLYDDPSQSSELLSEAKKLNDSQAPK(2)(2)

[0163] The anti-HER3 affibody includes the amino acid sequence shown in (3) known as Z (HER3: Z08699).

[0164] AEAKYAKEKYNAYYEIWQLPNLTKYQKAAFIGKLQDDPSQSSELLSEAKKLNDSQAPK(3)(3)

[0165] As anti-EGFR affibodies, they include the sequence shown in the amino acid sequence (4) known as Z (EGFR: 1907), the sequence shown in the amino acid sequence (5) known as Z (EGFR: 2377), the sequence shown in the amino acid sequence (6) known as Z (EGFR: 03115), the affibody molecule used for ABY-029, the sequence shown in the amino acid sequence (7) known as Z (PD-L1: 18609), or the sequence shown in the amino acid sequence (8) known as Z (PD-L1: 18608).

[0166] VDNKFNKEMWAAWEEIRNLPNLNGWQMTAFIASLVDDPSQSANLLAEAKKLNDAQAPK(4)(4)

[0167] VDNKFNKEMWIAWEEIRDLPNLNGWQMTAFIASLLDDPSQSANLLAEAKKLNDAQAPK(5)(5)

[0168] AEAKYAKEMWIAWEEIRNLPNLNGWQMTAFIAKLLDDPSQSSELLSEAKKLNDSQAPK(6)(6)

[0169] AEAKYAKERNAAAYEILYLPNLTNAQKWAFIWKLDDDPSQSSELLSEAKKLNDSQAPK(7)(7)

[0170] AEAKYAKERNKAAYEILYLPNLTNAQKWAFIWKLDDDPSQSSELLSEAKKLNDSQAPK(8)(8)

[0171] As a modification within the range of maintaining binding specificity, one or more amino acids can be used for modification, for example, 6×His, 3×HE tags or VDC, and an affibody containing these sequences can be separated and purified by the same steps as the method for recovering tag-labeled proteins.

[0172] Alternatively, these affibody molecules may be bound to copolymer X via a Cys residue or an amino group of an amino acid in the sequence, respectively.

[0173] When the compound of the present invention exists in geometrical isomers or optical isomers, mixtures or separated products of these isomers are also included in the scope of the present invention. Separation of isomers can be carried out according to conventional methods.

[0174] The target-specific copolymer or drug conjugate of the present invention can be produced by various known methods. The production method is not particularly limited, and for example, the target-specific copolymer or drug conjugate can be produced by the click reaction synthesis method described below.

[0175]

[0176] [wherein, R' represents a hydrogen atom or C 1-3 Alkyl, R" represents the above R 4 , R 5 or R 6 The groups shown.]

[0177] When the complexation of the targeting specific molecule and SCNP is implemented by covalent bonding, a functional group for complexation reaction can be introduced on the surface of the SCNP, and then complexation is carried out by reacting with the SH group, amino group or carboxyl group of the end or side chain of the targeting specific molecule that can react with the functional group. In addition, a spacer can be introduced at the end or side chain of the targeting specific molecule using an appropriate cross-linking reagent (cross-linking agent) to form a covalent bond with the SCNP in the form of a modified targeting specific molecule. As examples of such a bond formation, click chemistry between an azide group and an alkyne, a reaction between a thiol group and a maleimide group, and a reaction between an amino group and a succinimide group can be cited.

[0178] "Click chemistry" is classified as a reaction similar to natural biochemical reactions with the following properties, with the following characteristics. It is a very efficient reaction that proceeds rapidly to high yields, and is also a very selective reaction that does not generate any byproducts at all (or almost) and tolerates multiple functional groups. It is also a reaction carried out under mild reaction conditions such as low temperature (or ambient) or in aqueous solution. This reaction can be carried out in water or in the following solvents: alcohols such as methanol, ethanol, 1-propanol, 2-propanol; ethers such as diethyl ether, tetrahydrofuran, 1,4-dioxane; aromatic hydrocarbons such as benzene, toluene, and xylene; halogenated hydrocarbons such as dichloromethane, chloroform, 1,2-dichloroethane; N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, acetonitrile, ethyl acetate, etc., preferably in water or using N,N-dimethylformamide as a solvent. In some embodiments, cyclooctyne is dibenzocyclooctyne (DBCO), difluorobenzocyclooctyne (DIFBO), biaryl azacyclooctyne ketone (BARAC), dibenzocyclooctyne (DIBO), difluorocyclooctyne (DIFO), monofluorocyclooctyne (MOFO), dimethoxy azacyclooctyne (DIMAC) or aryl-less octyne (ALO), preferably dibenzocyclooctyne (DBCO). In some embodiments, the alkyne is an aliphatic alkyne, and the reaction step is carried out in the presence of a copper (I) catalyst. In some embodiments, the alkyne is cyclooctyne, and the reaction step is carried out under copper-free conditions. The reaction temperature is 0 to 300°C, preferably 0 to 150°C, more preferably 1 to 100°C, and the reaction time is 1 minute to 48 hours, preferably 5 minutes to 24 hours.

[0179] The reaction of the above reaction formula is a Huisgencycloaddition reaction as a click reaction, which is a 1,3-dipolar addition cyclization reaction of azide and alkyne to form 1,2,3-triazole.

[0180] The obtained polymer X, target-specific copolymer and drug complex of the present invention can be purified by separation and purification methods of polymers generally known in the field of polymer chemistry. Specific examples include: extraction, recrystallization, salting out using ammonium sulfate or sodium sulfate, centrifugal separation, dialysis, ultrafiltration, adsorption chromatography, ion exchange chromatography, hydrophobic chromatography, normal phase chromatography, reverse phase chromatography, desalting column chromatography, gel filtration, gel permeation chromatography, affinity chromatography, electrophoresis, countercurrent distribution, or a combination thereof. In particular, in hydrophobic chromatography, since the retention time varies depending on the number of SCNPs bound to each target-specific molecule, components that meet any DAR can be recovered by performing separation and extraction.

[0181] The copolymer X and the targeting specific copolymer of the present invention can be used as carriers for delivering various physiologically active substances (drugs). For example, the pharmaceutical composition formed by the copolymer X, the targeting specific copolymer or the drug complex of the present invention carrying (encapsulating) a tumor therapeutic drug can inhibit the proliferation of tumors as confirmed in the test examples described below, and can therefore be used as a preventive and / or therapeutic agent for various cancer diseases such as colorectal cancer, duodenal cancer, gastric cancer, pancreatic cancer, liver cancer, lung cancer, uterine cancer, and ovarian cancer. In addition, due to the high tumor accumulation ability, it can be used as a diagnostic agent and contrast agent for tumors.

[0182] When the copolymer and the target-specific copolymer of the present invention are used as drug delivery carriers, the dosage and the number of administrations can be appropriately selected in consideration of the dosage form, the patient's age, weight, the nature or severity of the symptoms to be treated, etc. The dosage and the number of administrations should not be limited. In the case of intravenous injection of a polymer or drug complex containing a drug, for example, it is preferred to administer 0.12 mg to 12,000,000 mg per adult (60 kg) in one administration, more preferably 1.2 mg to 1,200,000 mg, and particularly preferably 12 to 120,000 mg.

[0183] The pharmaceutical composition of the present invention can be prepared by mixing the copolymer X of the present invention, the targeting-specific copolymer or the drug complex with a drug. Preferably, the copolymer of the present invention, the targeting-specific copolymer or the drug complex is mixed with a drug to prepare single-chain nanoparticles, or the drug is mixed after preparing the single-chain nanoparticles of the copolymer X of the present invention, the targeting-specific copolymer or the drug complex. The single-chain nanoparticles can be prepared by a known method.

[0184] In the pharmaceutical composition of the present invention, the drug can be carried on the copolymer X, the targeting-specific copolymer or the drug complex by electrostatic interaction, hydrogen bonding, hydrophobic interaction or covalent bonding.

[0185] The drug is preferably an anticancer agent, more preferably an anticancer agent that acts on cancer cells to inhibit cancer cell proliferation, and examples thereof include metabolic antagonists, alkylating agents, anthracyclines, antibiotics, mitotic inhibitors, topoisomerase inhibitors, proteasome inhibitors, and antihormonal agents.

[0186] Examples of metabolic antagonists include azathioprine, 6-mercaptopurine, 6-thioguanine, fludarabine, pentostatin, cladribine, 5-fluorouracil (5FU), floxuridine (FUDR), cytarabine, methotrexate, trimethoprim, pyrimethamine, and pemetrexed. Examples of the alkylating agent include cyclophosphamide, mechlorethamine, uramustine, melphalan, chlorambucil, thiotepa / chlorambucil, ifosfamide, carmustine, lomustine, streptozocin, busulfan, dibromomannitol, cisplatin, carboplatin, nedaplatin, oxaliplatin, miriplatin, satraplatin, triplatin tetranitrate, procarbazine, altretamine, dacarbazine, mitozolamide, trabectedin, and temozolomide. Examples of anthracyclines include daunorubicin, doxorubicin, epirubicin, idarubicin, valrubicin, aclarubicin, amrubicin, pirarubicin, etc. Examples of antibiotics include actinomycin, bleomycin, mithramycin, anthramycin, streptozotocin, gramicidin D, staurosporine, mitomycins (e.g., mitomycin C), duocarmycins (e.g., CC-1065), and calicheamicins.Examples of the mitotic inhibitor include maytansinoids (e.g., DMO, maytansine (also known as DM1), DM2, DM3, DM4 or emtansine), auristatins (e.g., auristatin E, auristatin phenylalanine phenylenediamine (AFP), monomethyl auristatin E, monomethyl auristatin D and monomethyl auristatin F), dolastatins, cryptophycins, vinca alkaloids (e.g., vincristine, vinblastine, vindesine, vinorelbine), taxanes (e.g., paclitaxel, docetaxel) and colchicines. As topoisomerase inhibitors, for example, irinotecan, topotecan, noginotecan, amsacrine, etoposide, teniposide, mizantrone, mitoxantrone, SN-38, exitecan, derunotecan, etc. As proteasome inhibitors, for example, peptidylboronic acid, carfilzomib, bortezomib, etc. As antihormonal agents, for example, fulvestrant, tamoxifen, toremifene, etc. When these drugs are made into the pharmaceutical composition of the present invention, one kind or a combination of multiple kinds can be used, and the drug can be carried on the copolymer in the form of a free body.

[0187] The administration route of the pharmaceutical composition of the present invention is ideally the most effective route for treatment, and can be administered in the form of oral administration preparations, injections, or non-oral administration preparations such as transdermal administration preparations, for example, non-oral administration such as arterial injection, intravenous injection, subcutaneous injection, intramuscular injection, and intraperitoneal injection is preferred, and arterial injection and intravenous injection are more preferred. The number of administrations is not limited, and for example, it can be administered once to multiple times per week on average.

[0188] Various preparations suitable for the route of administration can appropriately select the excipients, extenders, binders, wetting agents, disintegrants, lubricants, surfactants, dispersants, buffers, preservatives, solubilizers, antiseptics, flavoring agents, analgesics, stabilizers, isotonic agents and other pharmaceutical additives commonly used in preparations, and can be manufactured according to conventional methods.

[0189] The preparation additives that the above-mentioned various preparations can contain are not particularly limited as long as they are pharmaceutically acceptable substances. Examples of such preparation additives include: purified water, water for injection, distilled water for injection, pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinyl pyrrolidone, carboxyvinyl polymers, sodium alginate, water-soluble dextran, sodium carboxymethyl starch, pectin, xanthan gum, gum arabic, casein, gelatin, agar, glycerol, propylene glycol, polyethylene glycol, vaseline, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, lactose, etc. The additives used can be appropriately selected according to various preparations and can be used alone or in combination.

[0190] In addition, the injection can also be made into a non-aqueous diluent (e.g., polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethanol, etc.), a suspending agent or an emulsion. The injection can be sterilized by filtration sterilization using a filter, a bactericide, etc. In addition, the injection can be made into a form prepared when used. That is, a sterile solid composition can be made by freeze drying, etc., and dissolved in water for injection, distilled water for injection, or other solvents before use.

[0191] Example

[0192] The present invention will be further specifically described below by way of examples. These examples are provided for illustration purposes and are not intended to limit the embodiments of the present invention.

[0193] [Example 1] Preparation of Poly[(Benzyl Acrylate)-(Poly(Ethylene Glycol) Methyl Ether Acrylate)-1-Ethoxyethyl Acrylate) Copolymer]

[0194] (1) Synthesis of 1-ethoxyethyl acrylate (EEA)

[0195] Ethyl vinyl ether (28.725 mL) was weighed under argon atmosphere, and phosphoric acid (50 mg) was added under ice cooling. Thereafter, acrylic acid (17.15 mL) was added, and the mixture was stirred at room temperature for 48 hours. Hydrotalcite (3 g) was added, and the mixture was further stirred for 2 hours to stop the reaction. After filtering with diatomaceous earth, unreacted ethyl vinyl ether was removed by evaporation. Phenothiazine was added as an inhibitor to make it 500 ppm, and the mixture was purified by vacuum distillation with calcium hydride (distillation temperature 28-32°C). The obtained 1-ethoxyethyl acrylate was divided into a glass vial and stored at -30°C.

[0196] 13C NMR (400MHz, CDCl3), δ, ppm: 15.29(-OCH2CH3), 21.16(-COOCH(CH3)), 64.98(-OCH2-), 96.73(-COOCH(CH3)), 128.84(CH2CH-), 131.43(CH2CH-), 166.00(-COO).

[0197] (2) Synthesis of Poly[(Benzyl acrylate)-(Poly(ethylene glycol) methyl ether acrylate)-(1-ethoxyethyl acrylate) Copolymer]

[0198] 100 mg of 2-(dodecylthiothiocarbonylthio)-2-methylpropionic acid (DDMAT) was weighed and dissolved in 17.3 mL of toluene to prepare a DDMAT / toluene stock solution (DDMAT concentration was 5.78 mg / mL). Similarly, 22 mg of 2,2'-azobis(2-methylpropionitrile) (AIBN) was weighed and dissolved in 17.3 mL of toluene to prepare an AIBN / toluene stock solution (AIBN concentration was 1.27 mg / mL). In addition, 1.296 g of poly(ethylene glycol) methyl ether acrylate (mPEGA, the average value (n) of the number of repeats of ethylene glycol was 9), 0.394 g of benzyl acrylate (BnA), 0.039 g of 1-ethoxyethyl acrylate, 1.73 mL of the DDMAT / toluene stock solution, and 1.73 mL of the AIBN / toluene stock solution were added, and polymerization was performed in an oil bath at 70°C. After 90 minutes, the polymerization was stopped, and the reaction solution was then subjected to a reprecipitation method or dialyzed against methanol to recover the copolymer. The obtained copolymer was basically a viscous body, so in the reprecipitation method, the reaction solution was added dropwise to a centrifuge tube to which a poor solvent (hexane / ethyl acetate = 7 / 3 [v / v]) was added, and recovered by centrifugation (2,000 × g, 5 min). This operation was repeated 3 times, and finally vacuum dried to obtain 1.223 g of poly[(benzyl acrylate)-(poly(ethylene glycol) methyl ether acrylate)-(1-ethoxyethyl acrylate) copolymer].

[0199] The obtained copolymer was measured by NMR. 1 The H-NMR spectrum shows the degree of polymerization and number average molecular weight (M n,NMR ) were analyzed, and the results showed that the degree of polymerization of mPEGA (n = 9) was 102, the degree of polymerization of BnA was 94, the degree of polymerization of EEA was 9, and the degree of polymerization of M n,NMR The molecular weight dispersion (M w / M n ), the result is 1.53.

[0200]

[0201] [Measurement equipment and conditions, etc.]

[0202] (1) 1 H-NMR measurement

[0203] Device: JNM-ECX400 (400MHz) / JEOL Ltd.

[0204] Solvent: Dimethyl sulfoxide-d6 containing 0.03% tetramethylsilane / Kanto Chemical Co., Ltd.

[0205] Sample concentration: 20mg / mL

[0206] Measurement temperature: 25℃

[0207] Total times: 256 times

[0208] result: Figure 1

[0209] (2) GPC measurement

[0210] Equipment: HPLC-Prominence system / Shimadzu Corporation

[0211] Detector: RID-10A refractive index detector / Shimadzu Corporation

[0212] Column: TSKgelα-2500 column / Tosoh Corporation

[0213] (column size 7.8 mm × 300 mm, particle size 7 μm, exclusion limit molecular weight 5 × 10 3 )

[0214] TSKgelα-4000 column / Tosoh Corporation

[0215] (column size 7.8 mm × 300 mm, particle size 10 μm, exclusion limit molecular weight 4 × 10 5 )

[0216] TSKgel guard column / Tosoh Corporation

[0217] Mobile phase: N,N-dimethylformamide (DMF) containing 10mmol / L lithium bromide

[0218] Temperature: 40℃

[0219] Flow rate: 0.5mL / min

[0220] Sample concentration: 6mg / mL

[0221] Standard material: Poly(methyl methacrylate) standard ReadyCal set, M p800-2,200,000Da / SIGMA

[0222] result: Figure 2

[0223] [Table 1]

[0224]

[0225] [Examples 2 to 68]

[0226] The types, addition amounts, reaction temperatures and polymerization times of the monomers (mPEGA, BnA and EEA) used in Example 1 were appropriately changed, and polymers having different composition ratios or average molecular weights as shown in the following table were prepared in the same manner as in Example 1.

[0227] [Table 2]

[0228]

[0229] [Table 3]

[0230]

[0231] [Example 69]

[0232] Production of poly[(benzyl acrylate)-(poly(ethylene glycol) methyl ether acrylate)-(acrylic acid) copolymer]

[0233] The poly[(benzyl acrylate)-(poly(ethylene glycol) methyl ether acrylate)-(1-ethoxyethyl acrylate) copolymer] obtained in Example 1 was treated with 0.5N HCl at room temperature to remove the ethoxyethyl group and obtain 1.176 g of a terpolymer having a carboxyl group. The Z average particle size and polydispersity index of the obtained terpolymer in water were measured by dynamic light scattering (DLS), and the result was 8.5 nm (polydispersity index 0.14).

[0234]

[0235] [Measurement equipment and conditions, etc.]

[0236] (1) DLS assay

[0237] Device: Zetasizer NanoZS / Malvern Instruments Ltd.

[0238] Measurement temperature: 25℃

[0239] Sample concentration: 10mg / mL

[0240] result: Figure 3

[0241] [Example 70]

[0242] Method for producing SCNPs encapsulated with (1,2-diaminocyclohexane) platinum (II)

[0243] Dissolve 65.28 mg of the Cl(H2O) form (DACHPt·Cl·H2O) of (1,2-diaminocyclohexane)platinum(II) (hereinafter abbreviated as DACHPt) in 20 mL of purified water and stir at 70°C for 2 hours. Add 287.4 mg of the terpolymer obtained in Example 69 to 5 mL of the solution and stir at 50°C overnight. After stirring, the reaction solution was dialyzed and purified using purified water as the external liquid to obtain 5 mL of DACHPt-encapsulated SCNPs. The Pt content of the purified DACHPt-encapsulated SCNPs was measured by inductively coupled plasma-atomic emission spectrometry (ICP-AES) and was 720 μg / mL (1.14 mg / mL in terms of DACHPt). Another 200 μL of DACHPt-encapsulated SCNPs was freeze-dried, and the solid content concentration was calculated. The ratio of the solid content to the Pt content was taken to calculate the Pt binding amount per unit polymer, and the result was 3.4 mol / mol. In addition, the Z average particle size and polydispersity index of the obtained DACHPt-encapsulated SCNPs were measured by dynamic light scattering (DLS), and the result was 8.7 nm (polydispersity index 0.14). The particle size of SCNPs before and after DACHPt encapsulation is shown in Figure 3 The particle size of SCNPs hardly changed before and after DACHPt encapsulation. The results are summarized in the table below.

[0244] [Measurement equipment and conditions, etc.]

[0245] (1) ICP-AES measurement

[0246] Device: Continuous high-frequency plasma light emitting device ICPE-9000 / Shimadzu Corporation

[0247] Pretreatment device: Microwave sample pretreatment device ETHOS EASY / Milestone General

[0248] Measurement wavelength: 214nm

[0249] Standard solution: Platinum standard solution (Pt1000) for ICP analysis / Fujifilm Wako Pure Chemical Industries, Ltd.

[0250] (2) DLS assay

[0251] Device: Zetasizer NanoZS / Malvern Instruments Ltd.

[0252] Measurement temperature: 25℃

[0253] Sample concentration: 10mg / mL

[0254] result: Figure 3

[0255] [Table 4]

[0256]

[0257] [Example 71]

[0258] Production of N3-poly[(benzyl acrylate)-(poly(ethylene glycol) methyl ether acrylate)-(1-ethoxyethyl acrylate) copolymer]

[0259] N3-CTA was used to replace the chain transfer agent DDMAT used in Example 1 for synthesis. 100 mg of 2-(dodecylthiothiocarbonylthio)-2-methylpropionic acid-3-azido-1-propanol ester (N3-CTA) was weighed and dissolved in 17.3 mL of toluene to prepare an N3-CTA / toluene stock solution (with an N3-CTA concentration of 5.78 mg / mL). Similarly, 10 mg of 2,2'-azobis(2-methylpropionitrile) (AIBN) was weighed and dissolved in 7.87 mL of toluene to prepare an AIBN / toluene stock solution (with an AIBN concentration of 1.27 mg / mL). 2.592 g of poly(ethylene glycol) methyl ether acrylate (mPEGA, the average value (n) of the number of repeats of ethylene glycol is 9), 0.700 g of benzyl acrylate (BnA), 0.156 g of 1-ethoxyethyl acrylate, 4.15 mL of N3-CTA / toluene stock solution, and 3.46 mL of AIBN / toluene stock solution were added, and polymerization was carried out in an oil bath at 70° C. After 90 minutes, the polymerization was stopped, and the reaction solution was then recovered by reprecipitation or dialysis against methanol to recover the copolymer. The obtained copolymer was basically a viscous body, so in the reprecipitation method, the reaction solution was added dropwise to a centrifuge tube to which a poor solvent (hexane / ethyl acetate = 7 / 3 [v / v]) was added, and the mixture was recovered by centrifugal separation (2,000×g, 5 min). This operation was repeated 3 times, and finally vacuum dried to obtain 2.643 g of N3-poly[(benzyl acrylate)-(poly(ethylene glycol) methyl ether acrylate)-(1-ethoxyethyl acrylate) copolymer. The obtained copolymer was measured by NMR. 1 The H-NMR spectrum shows the degree of polymerization and number average molecular weight (Mn,NMR ) were analyzed, and the results showed that the polymerization degree of mPEGA (n = 9) was 75, the polymerization degree of BnA was 55, the polymerization degree of EEA was 26, and the polymerization degree of M n,NMR The molecular weight dispersion (M w / M n ), the result is 1.28.

[0260] [Example 72]

[0261] Preparation of N3-poly[(benzyl acrylate)-(poly(ethylene glycol) methyl ether acrylate)-(acrylic acid) copolymer]

[0262] The N3-poly[(benzyl acrylate)-(poly(ethylene glycol) methyl ether acrylate)-(1-ethoxyethyl acrylate) copolymer] obtained in Example 71 was treated with 0.5N HCl at room temperature to remove the ethoxyethyl group and obtain 2.35 g of N3-poly[(benzyl acrylate)-(poly(ethylene glycol) methyl ether acrylate)-(acrylic acid) copolymer] having a carboxyl group. The Z average particle size and polydispersity index of the obtained copolymer in water were measured by dynamic light scattering (DLS), and the result was 8.0 nm (polydispersity index 0.13).

[0263] [Measurement equipment and conditions, etc.]

[0264] (1) 1 H-NMR measurement

[0265] Device: JNM-ECX400 (400MHz) / JEOL Ltd.

[0266] Solvent: Dimethyl sulfoxide-d6 containing 0.03% tetramethylsilane / Kanto Chemical Co., Ltd.

[0267] Sample concentration: 20mg / mL

[0268] Measurement temperature: 25℃

[0269] Total times: 256 times

[0270] result: Figure 4

[0271] (2) GPC measurement

[0272] Equipment: HPLC-Prominence system / Shimadzu Corporation

[0273] Detector: RID-10A refractive index detector / Shimadzu Corporation

[0274] Column: TSKgelα-2500 column / Tosoh Corporation

[0275] (column size 7.8 mm × 300 mm, particle size 7 μm, exclusion limit molecular weight 5 × 10 3 )

[0276] TSKgelα-4000 column / Tosoh Corporation

[0277] (column size 7.8 mm × 300 mm, particle size 10 μm, exclusion limit molecular weight 4 × 10 5 )

[0278] TSKgel guard column / Tosoh Corporation

[0279] Mobile phase: N,N-dimethylformamide (DMF) containing 10mmol / L lithium bromide

[0280] Temperature: 40℃

[0281] Flow rate: 0.5mL / min

[0282] Sample concentration: 6mg / mL

[0283] Standard material: Poly(methyl methacrylate) standard ReadyCal set, M p 800-2,200,000Da / SIGMA

[0284] result: Figure 5

[0285] [Table 5]

[0286]

[0287] [Example 73]

[0288] Synthesis of 1,4-diaminobutane-conjugated copolymers

[0289] 2350 mg of the copolymer obtained in Example 72 was weighed and dissolved in 47 mL of DMF, and 1111.1 mg of COMU and 527 μL of TMP were added, and the mixture was stirred at room temperature for 3 hours. Then, 2512 μL of N-(tert-butoxycarbonyl)-1,4-diaminobutane was added, and the mixture was stirred at 30°C for 3 days. The reaction solution was purified by dialysis (dialysis membrane: Spectra / Por Regenerated CelluloseMembrane 6, molecular weight cutoff: 3.5 kDa, external solution: methanol), and the solvent was removed by vacuum distillation and vacuum drying to recover the copolymer. The obtained N-Boc-1,4-diaminobutane-bound copolymer was dissolved in 32 mL of a mixed solution of DCM and TFA [DCM / TFA=5 / 3 (v / v)], and the mixture was stirred at room temperature overnight to perform deprotection, and the solvent was removed by vacuum distillation and vacuum drying to obtain 2517 mg of 1,4-diaminobutane-bound copolymer.

[0290]

[0291] For 1,4-diaminobutane-bound copolymers, the 1 The H-NMR spectrum analysis revealed that the amount of 1,4-diaminobutane introduced per molecule of the copolymer was 26 mol / mol.

[0292] [Measurement equipment and conditions, etc.]

[0293] (1) 1 H-NMR measurement

[0294] Device: JNM-ECX400 (400MHz) / JEOL Ltd.

[0295] Solvent: Dimethyl sulfoxide-d6 containing 0.03% tetramethylsilane / Kanto Chemical Co., Ltd.

[0296] Sample concentration: 20mg / mL

[0297] Measurement temperature: 25℃

[0298] Total times: 256 times

[0299] result: Figure 6

[0300] [Table 6]

[0301] Example Copolymers used Number of 1,4-diaminobutane introduced per molecule of copolymer (mol / mol) 73 Embodiment 72 26

[0302] [Example 74]

[0303] Preparation of DM1-SPDP-1,4-diaminobutane conjugate copolymer

[0304] 81 mg of maytansine (DM1) and 29 mg of 2,5-dioxopyrrolidin-1-yl 3-(pyridin-2-yldisulfaneyl) propanoate (SPDP) were added and dissolved in 1 mL of DCM, 16 μL of N,N-diisopropylethylamine (DIPEA) was added, and stirred at 30° C. for 3 hours. 200 mg of the copolymer obtained in Example 73 was dissolved in 4 mL of DCM and added to the reaction solution, and the resulting reaction solution was stirred at 30° C. for 24 hours. The reaction solution was purified by dialysis (dialysis membrane: Spectra / Por Regenerated CelluloseMembrane 6, molecular weight cutoff: 3.5 kDa, external solution: methanol), and the solvent was removed by vacuum distillation and vacuum drying to recover the copolymer to obtain 189 mg of DM1-SPDP-1,4-diaminobutane-bound copolymer. The Z average particle size and polydispersity index of the obtained copolymer in water were measured by dynamic light scattering (DLS), and the result was 13 nm (polydispersity index 0.23).

[0305]

[0306] For DM1-SPDP-1,4-diaminobutane conjugate copolymers, the 1 The amount of DM1 introduced per molecule of the copolymer was analyzed by H-NMR spectrum and found to be 26 mol / mol.

[0307] [Measurement equipment and conditions, etc.]

[0308] (1) 1 H-NMR measurement

[0309] Device: JNM-ECX400 (400MHz) / JEOL Ltd.

[0310] Solvent: Dimethyl sulfoxide-d6 containing 0.03% tetramethylsilane / Kanto Chemical Co., Ltd.

[0311] Sample concentration: 20mg / mL

[0312] Measurement temperature: 25℃

[0313] Total times: 256 times

[0314] result: Figure 7

[0315] [Table 7]

[0316] Example Copolymers used Number of DM1 introduced per molecule of copolymer (mol / mol) 74 Embodiment 73 26

[0317] [Example 75]

[0318] Manufacturing of anti-HER2 affibody micelle drug complexes

[0319] The copolymer obtained in Example 74 was complexed with the anti-HER2 affibody. 2 μL of 0.5 mM tris(2-carboxyethyl)phosphine (TCEP) dissolved in PBS was added to a glass vial containing 14 mL of a stock solution (0.53 mg / mL) of the anti-Her2 affibody (Z (HER2: 2891) containing a 6×His tag, amino acid sequence number (9)), and the mixture was stirred at 37°C for 15 minutes. Then, 1.4 mg of DBCO-PEG4-maleimide was dissolved in 1 mL of DMAc (N,N-dimethylacetamide) and added, and the mixture was stirred at 37°C for 15 minutes to react with the terminal cysteine ​​residue of the affibody. 20 μL of an aqueous solution (10 mg / mL) of L-cysteine ​​(168149, Sigma-Aldrich) dissolved in PBS was added, and the mixture was stirred at 37°C for 1 minute to stop the reaction. The reaction solution was recovered and dialyzed against purified water overnight (Slide-A-LyzerTM Dialysis Cassette, molecular weight cutoff: 3.5K, 66110, Thermo Fisher Scientific) and ultrafiltered (Vivaspin Turbo 15, molecular weight cutoff: 3K) to remove the unreacted modification reagent. The volume was precisely determined to 5 mL using PBS, and 1 mL of the solution was dispensed into a glass vial. Next, 60 mg of the copolymer obtained in Example 74 was added and the mixture was stirred at 4 °C for 2 h. TM The mixture was allowed to stand at -20°C for 1 day. Thereafter, the solution was thawed at 4°C and recovered. In anion exchange chromatography, the copolymer-bound affibody was separated from the copolymer (raw material) or free affibody (raw material) alone according to the difference in retention time, and then washed with ultrafiltration (Vivaspin Turbo15, molecular weight cutoff: 10K) using PBS and concentrated. Next, immobilized metal affinity chromatography (IMAC: immobilized metal ion affinity chromatography) was performed with the His-tagged protein expressed in the Z (HER2: 2891) affibody containing a 6×His tag as the target to obtain the anti-HER2 affibody micelle drug complex as the target. The Z average particle size and polydispersity index of the obtained anti-HER2 affibody micelle drug complex in water were measured by dynamic light scattering (DLS), and the result was 11 nm (polydispersity index 0.17).

[0320]

[0321] [Measurement equipment and conditions, etc.]

[0322] (1) Anion exchange column purification

[0323] Device: AKTA primePlus / Cytiva

[0324] Detector: UV / 280nm

[0325] Column: HiTrap Q XL (1mL) / Cytiva

[0326] Mobile phase A: 20mmol / LTris buffer (pH 10)

[0327] Mobile phase B: 20mmol / LTris buffer (pH 10) + 1M NaCl (pH 10)

[0328] Temperature: 25℃

[0329] Flow rate: 1.0mL / min

[0330] Sample loop: 0.5mL

[0331] (2) IMAC purification

[0332] Device: AKTA primePlus / Cytiva

[0333] Detector: UV / 280nm

[0334] Column: HisTrap HP (5mL) / Cytiva

[0335] Mobile phase A: 20mmol / L trisodium phosphate buffer + 150mmol / L NaCl (pH 7.6)

[0336] Mobile phase B: 20mmol / L trisodium phosphate buffer + 500mmol / L NaCl + 500mmol / L imidazole (pH 7.6)

[0337] Temperature: 25℃

[0338] Flow rate: 2.0mL / min

[0339] Sample loop: 0.5mL

[0340] (3) DLS measurement

[0341] Device: Zetasizer NanoZS / Malvern Instruments Ltd.

[0342] Measurement temperature: 25℃

[0343] Sample concentration: 10mg / mL

[0344] result: Figure 8

[0345] [Example 76]

[0346] Preparation of anti-EGFR affibody micelle drug complex

[0347] The copolymer obtained in Example 74 and the anti-EGFR affibody (Z (EGFR: 03115) containing a 6×His tag, amino acid sequence number (10)) were complexed in the same manner as in Example 75. The Z average particle size and polydispersity index of the anti-EGFR affibody micelle drug complex obtained in water were measured by dynamic light scattering, and the result was 14 nm (polydispersity index 0.19).

[0348] [Measurement equipment and conditions, etc.]

[0349] (1) Anion exchange column purification

[0350] Device: AKTA primePlus / Cytiva

[0351] Detector: UV / 280nm

[0352] Column: HiTrap Q XL (1mL) / Cytiva

[0353] Mobile phase A: 20mmol / LTris buffer (pH 10)

[0354] Mobile phase B: 20mmol / LTris buffer (pH 10) + 1M NaCl (pH 10)

[0355] Temperature: 25℃

[0356] Flow rate: 1.0mL / min

[0357] Sample loop: 0.5mL

[0358] (2) IMAC purification

[0359] Device: AKTA primePlus / Cytiva

[0360] Detector: UV / 280nm

[0361] Column: HisTrap HP (5mL) / Cytiva

[0362] Mobile phase A: 20mmol / L trisodium phosphate buffer + 150mmol / L NaCl (pH 7.6)

[0363] Mobile phase B: 20mmol / L trisodium phosphate buffer + 500mmol / L NaCl + 500mmol / L imidazole (pH 7.6)

[0364] Temperature: 25℃

[0365] Flow rate: 2.0mL / min

[0366] Sample loop: 0.5mL

[0367] (3) DLS measurement

[0368] Device: Zetasizer NanoZS / Malvern Instruments Ltd.

[0369] Measurement temperature: 25℃

[0370] Sample concentration: 10mg / mL

[0371] result: Fig. 9

[0372] [Example 77]

[0373] Preparation of anti-HER3 affibody micelle drug complex

[0374] The copolymer obtained in Example 74 and the anti-HER3 affibody (Z (HER3: Z08699) containing a 3×HE tag, amino acid sequence number (11)) were complexed in the same manner as in Example 75. The Z average particle size and polydispersity index of the anti-HER3 affibody micelle drug complex obtained in water were measured by dynamic light scattering, and the result was 11 nm (polydispersity index 0.15).

[0375] [Measurement equipment and conditions, etc.]

[0376] (1) Anion exchange column purification

[0377] Device: AKTA primePlus / Cytiva

[0378] Detector: UV / 280nm

[0379] Column: HisTrap Q XL (1 mL) / Cytiva

[0380] Mobile phase A: 20mmol / LTris buffer (pH 10)

[0381] Mobile phase B: 20mmol / LTris buffer (pH 10) + 1M NaCl (pH 10)

[0382] Temperature: 25℃

[0383] Flow rate: 1.0mL / min

[0384] Sample loop: 0.5mL

[0385] (2) IMAC purification

[0386] Device: AKTAprimePlus / Cytiva

[0387] Detector: UV / 280nm

[0388] Column: HisTrap HP (5mL) / Cytiva

[0389] Mobile phase A: 20mmol / L trisodium phosphate buffer + 150mmol / L NaCl (pH 7.6)

[0390] Mobile phase B: 20mmol / L trisodium phosphate buffer + 500mmol / L NaCl + 500mmol / L imidazole (pH 7.6)

[0391] Temperature: 25℃

[0392] Flow rate: 2.0mL / min

[0393] Sample loop: 0.5mL

[0394] (3) DLS measurement

[0395] Device: Zetasizer NanoZS / Malvern Instruments Ltd.

[0396] Measurement temperature: 25℃

[0397] Sample concentration: 10mg / mL

[0398] result: Fig.10

[0399] [Comparative Example 1]

[0400] Preparation of oxaliplatin solution

[0401] 1 mL of ELPLAT intravenous injection solution 50 mg (Yakult Honsha Co., Ltd.) was added to 5.58 mL of 5.9 (w / v)% glucose solution to prepare a 5 (w / v)% glucose solution containing 760 μg of oxaliplatin.

[0402] [Comparative Example 2]

[0403] Preparation of DM1 solution

[0404] Maytansine (DM1) (Cayman CHEMICAL) was dissolved in dimethyl sulfoxide (DMSO, Fujifilm Wako Pure Chemical Industries, Ltd.) to prepare solutions with a concentration of 60 μmol / L, 20 μmol / L, 6.7 μmol / L, 2.2 μmol / L, 0.74 μmol / L, 0.25 μmol / L, 0.082 μmol / L or 0.027 μmol / L of DM1. Next, these solutions were diluted using RPMI1640 (hereinafter referred to as RPMI1640 medium) containing 10% fetal bovine serum (Merck KGaA) to prepare a dilution series of 60nmol / L, 20nmol / L, 6.7nmol / L, 2.2nmol / L, 0.74nmol / L, 0.25nmol / L, 0.082nmol / L or 0.027nmol / L, and then they were added to the cell culture microplate in an amount equal to the cell culture medium and diluted to one-half, so that the final concentration of DM1 was 30nmol / L, 10nmol / L, 3.3nmol / L, 1.1nmol / L, 0.37nmol / L, 0.12nmol / L, 0.041nmol / L or 0.014nmol / L to evaluate the anti-cellular effect.

[0405] [Comparative Example 3]

[0406] Preparation of Z (HER2: 2891) solution containing 6×His tag

[0407] Z (HER2: 2891) containing a 6×His tag as an anti-HER2 affibody (homemade, a physiological saline solution with a concentration of 0.393 mg / mL as anti-HER2 affibody, amino acid sequence number (9)) was diluted using RPMI1640 medium to prepare 600 ng / mL, 200 ng / mL, 60 ng / mL, 20 ng / mL, 6.0 ng / mL, 2.0 ng / mL, 0.60 ng / mL, 0.2 0 ng / mL or 0.060 ng / mL dilution series, and then they were added to the cell culture microplate in an amount equal to the cell culture medium, diluted to one-half, so that the anti-HER2 affibody had a final concentration of 300 ng / mL, 100 ng / mL, 30 ng / mL, 10 ng / mL, 3.0 ng / mL, 1.0 ng / mL, 0.30 ng / mL, 0.10 ng / mL or 0.030 ng / mL to evaluate the anti-cell effect.

[0408] [Comparative Example 4]

[0409] Preparation of Z (EGFR: 03115) solution containing 6×His tag

[0410] Z (EGFR: 03115) containing a 6×His tag as an anti-EGFR affibody (homemade, a physiological saline solution with a concentration of 0.101 mg / mL as anti-EGFR affibody, amino acid sequence number (10)) was diluted in RPMI1640 medium to prepare 600 ng / mL, 200 ng / mL, 60 ng / mL, 20 ng / mL, 6.0 ng / mL, 2.0 ng / mL, 0.60 ng / mL, 0. The anti-EGFR affibody was prepared by diluting the anti-EGFR affibody in a series of dilutions of 20 ng / mL or 0.060 ng / mL, and then adding them to the cell culture microplate in an amount equal to the cell culture medium, diluted to one-half, so that the anti-cell effect was evaluated at a final concentration of 300 ng / mL, 100 ng / mL, 30 ng / mL, 10 ng / mL, 3.0 ng / mL, 1.0 ng / mL, 0.30 ng / mL, 0.10 ng / mL or 0.030 ng / mL.

[0411] [Test Example 1] Drug efficacy test

[0412] Mouse colon cancer cell line C26 (American Type Culture Collection) was subcutaneously transplanted into female nude mice (BALB / c-nu / nu, 7 weeks old; Charles River Laboratories Japan, Inc.), and the resulting cancer-bearing model was used for drug efficacy testing.

[0413] Mouse colorectal cancer cell line C26 subcultured in a CO2 incubator was suspended in liquid culture medium (Dulbecco's Modified Eagle's Medium-high glucose, Sigma-Aldrich) at a cell number of 1×10 6 The nude mice were then subcutaneously injected with 100 μL of the drug. After the mice were fed for about a week, the tumors grew to an average volume of about 30 mm. 3 The drug was started at 1:00 p.m. DACHPt-encapsulated SCNP (DACHPt-encapsulated SCNP prepared using the copolymer of Example 70) was administered intravenously in the tail vein (3 times every other day), and the anti-tumor effect was evaluated based on the tumor volume (4 to 5 mice in one group). For comparison, an oxaliplatin solution (Comparative Example 1) was used and administered in the same manner. Regarding the dosage of each preparation, the maximum dosage of the oxaliplatin solution is 8 mg / kg (3.9 mg / kg in terms of Pt), and the DACHPt-encapsulated SCNP is 3 mg / kg in terms of Pt.

[0414] The changes in tumor volume over time are shown in Fig.11 . In the case of DACHPt containing SCNPs, T / C = 0.4 [T / C: ratio of tumor volume of drug administration group (T) to control group (C)] after 14 days of administration. In the case of oxaliplatin solution (Comparative Example 1), T / C = 1.1 after 14 days of administration. In addition, it was confirmed that DACHPt containing SCNPs significantly inhibited tumor growth compared to the control group after 14 days of administration (Student t test). The above results indicate that DACHPt containing SCNPs has an excellent anti-tumor effect compared to oxaliplatin solution.

[0415] [Test Example 2] Anti-cell test

[0416] The HER2 antigen-positive human gastric cancer cell line HCC-1954 (American Type Culture Collection) or the HER2 antigen-negative human breast cancer cell line MDA-MB-468 (American Type Culture Collection) were adjusted to 2.0×10 4 cells / mL, 100 μL was added to each 96-well cell culture microplate and cultured overnight. The next day, 100 μL of the evaluation specimen diluted with RPMI1640 medium was added to each microplate and cultured for 4 days at 37°C and 5% CO2. After culture, CellTiter-Glo was added to the microplate. TMLuminescent Cell Viability Assay (Promega Corporation) was used and stirred, and after standing at room temperature for 10 minutes, the luminescence intensity was measured using a plate reader (Molecular Devices, LLC.) The cell viability was calculated according to the following formula.

[0417] Cell survival rate (%) = a ÷ b × 100

[0418] [wherein: a represents the average value of the luminescence amount of the sample wells (n=3), and b represents the average value of the luminescence amount of the wells without adding the sample (n=3)]

[0419] The IC concentration at which cell viability reaches 50% 50 Values ​​using SAS TM 9.4Software (SAS Institute Japan Ltd.) Calculated.

[0420] [Table 8]

[0421]

[0422] EGFR antigen-positive human breast cancer cell line MDA-MB-468 (American Type Culture Collection) or EGFR antigen-negative human breast cancer cell line MDA-MB-453 (RIKEN BRC CELL BANK) were adjusted to 2.0×10 4 cells / mL and 4.0×10 4 Cells / mL, 100 μL of each were added to a 96-well cell culture microplate and cultured overnight. The next day, 100 μL of the evaluation specimen diluted with RPMI 1640 medium was added to each microplate and cultured for 4 days at 37°C and 5% CO2. After culture, CellTiter-Glo was added to the microplate. TM Luminescent Cell Viability Assay (Promega Corporation) was used and stirred. After standing at room temperature for 10 minutes, the luminescence was measured using a plate reader (Molecular Devices, LLC.). Cell viability and IC 50 value.

[0423] [Table 9]

[0424]

Claims

1. A copolymer formed by combining a targeting specific molecule with a copolymer X having a structural unit represented by the following formulae (A), (B) and (C), wherein: In the formula, R 1 , R 2 and R 3 Same or different, indicating hydrogen atoms or C 1-3 Alkyl; R 4 Represents C 1-3 Alkyl; R 5 represents hydrogen atom, C 1-18 alkyl, 3- to 8-membered cycloalkyl which may have a substituent, adamantyl, C 6-18 Aryl or 5- to 10-membered heteroaryl which may have a substituent; X 1 , X 2 and X 3 The same or different, representing oxygen atoms, sulfur atoms or NR 7 ; R 6 represents a hydrogen atom, a leaving group or a connecting group; R 7 Represents a hydrogen atom or C 1-3 An alkyl group; m represents an integer of 1 to 100; and n represents an integer of 0 to 3.

2. The copolymer according to claim 1, characterized in that The polymer X is a copolymer formed by polymerizing three monomers represented by the following general formulas (1) to (3): In the formula, R 1 , R 2 and R 3 Same or different, indicating hydrogen atoms or C 1-3 Alkyl; R 4 Represents C 1-3 Alkyl; R 5 represents hydrogen atom, C 1-18 alkyl, 3- to 8-membered cycloalkyl which may have a substituent, adamantyl, C 6-18 Aryl or 5- to 10-membered heteroaryl which may have a substituent; X 1 , X 2 and X 3 The same or different, representing oxygen atoms, sulfur atoms or NR 7 ; R 6 represents a hydrogen atom, a leaving group or a connecting group; R 7 Represents a hydrogen atom or C 1-3 An alkyl group; m represents an integer of 1 to 100; and n represents an integer of 0 to 3.

3. The copolymer according to claim 1 or 2, characterized in that R 1 A hydrogen atom.

4. The copolymer according to claim 1 or 2, characterized in that R 2 A hydrogen atom.

5. The copolymer according to claim 1 or 2, characterized in that R 3 A hydrogen atom.

6. The copolymer according to claim 1 or 2, characterized in that R 4 It is methyl.

7. The copolymer according to claim 1 or 2, characterized in that R 5 is a C which may have a substituent 6-18 Aryl.

8. The copolymer according to claim 1 or 2, characterized in that R 5 It is phenyl.

9. The copolymer according to claim 1 or 2, characterized in that R 6 A hydrogen atom.

10. The copolymer according to claim 1 or 2, characterized in that R 6 The leaving group is a group represented by the following formula (4):

11. The copolymer according to claim 1 or 2, characterized in that R 6 The connecting group is a group represented by the following formula (5): In the formula, R 8 Represents a hydrogen atom or a drug; Ak 1 Represents C 1-7 Alkylene bond; X 4 represents an oxygen atom, a sulfur atom or -N(R 7 )-, where R 7 Represents a hydrogen atom or C 1-3 alkyl.

12. The copolymer according to claim 1 or 2, characterized in that X 1 For oxygen atoms.

13. The copolymer according to claim 1 or 2, characterized in that X 2 For oxygen atoms.

14. The copolymer according to claim 1 or 2, characterized in that X 3 is an oxygen atom or NH.

15. The copolymer according to claim 1 or 2, characterized in that m is an integer from 4 to 22.

16. The copolymer according to claim 1 or 2, characterized in that n is 1.

17. The copolymer according to claim 1, characterized in that The ratio of the structural units (A), (B), and (C) is 0.01 to 100 parts by mass of (B) and 0.1 to 100 parts by mass of (C) per 1 part by mass of (A).

18. The copolymer according to claim 2, characterized in that The monomer (2) and the monomer (3) are polymerized in an amount of 0.01 to 100 parts by mass based on 1 part by mass of the monomer (1).

19. The copolymer according to claim 1 or 2, characterized in that The number average molecular weight is 5000 to 150000.

20. The copolymer according to claim 1 or 2, characterized in that The target-specific molecule is an affibody.

21. The copolymer according to claim 20, characterized in that The affinity body is an anti-HER2 affinity body, an anti-HER3 affinity body, an anti-EGFR affinity body, an anti-PD-L1 affinity body, an anti-TNFα affinity body, an anti-fibrinogen affinity body, an anti-transferrin affinity body, an anti-HSA affinity body, an anti-insulin affinity body, an anti-IgG affinity body, an anti-IgM affinity body, an anti-IgA affinity body or an anti-IgE affinity body.

22. The copolymer according to claim 20, characterized in that The affibody is a molecule comprising the amino acid sequences (2) to (6): AEAKYAKEMRNAYWEIALLPNLTNQQKRAFIRKLYDDPSQSSE LLSEAKKLNDSQAPK(sequence number 2)(2) AEAKYAKEKYNAYYEIWQLPNLTKYQKAAFIGKLQDDPSQSSE LLSEAKKLNDSQAPK(Serial No. 3)(3) VDNKFNKEMWAAWEEIRNLPNLNGWQMTAFIASLVDDPSQSA NLLAEAKKLNDAQAPK(Sequence No. 4)(4) VDNKFNKEMWIAWEEIRDLPNLNGWQMTAFIASLLDDPSQSA NLLAEAKKLNDAQAPK(sequence number 5)(5) AEAKYAKEMWIAWEEIRNLPNLNGWQMTAFIAKLLDDPSQSS ELLSEAKKLNDSQAPK(Serial No. 6)(6) AEAKYAKERNAAAYEILYLPNLTNAQKWAFIWKLDDDPSQSSE LLSEAKKLNDSQAPK(sequence number 7)(7) AEAKYAKERNKAAYEILYLPNLTNAQKWAFIWKLDDDPSQSSELLSEAKKLNDSQAPK (sequence number 8) (8).

23. A drug complex, characterized in that The invention comprises the copolymer according to claim 1 or 2 and a drug.

24. The drug complex according to claim 23, wherein The drug is a metabolic antagonist, an alkylating agent, an anthracycline, an antibiotic, a mitotic inhibitor, a topoisomerase inhibitor, a proteasome inhibitor or an anti-hormone agent.

25. The drug complex according to claim 23, characterized in that The drug is DM0, DM1, DM2, DM3, DM4, metansine, auristatin E, auristatin phenylalanine phenylenediamine (AFP), monomethyl auristatin E, monomethyl auristatin D, monomethyl auristatin F, paclitaxel, docetaxel, irinotecan, topotecan, nojitecan, amsacrine, etoposide, teniposide, mitoxantrone, SN-38, exotecan or derunotecan.

26. The drug complex according to claim 23, characterized in that The binding between the target specific molecule or drug and the copolymer X is covalent binding or non-covalent binding.

27. A single-chain nanoparticle, characterized in that The copolymer comprising claim 1 or 2.

28. A single-chain nanoparticle, characterized in that Comprising the drug complex of claim 23.

29. A pharmaceutical composition, characterized in that The copolymer comprising claim 1 or 2.

30. A pharmaceutical composition, characterized in that A drug complex comprising the drug complex of claim 23.

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