Particle for drug delivery to heart and / or diaphragm and pharmaceutical composition

Abstract

A main object of the present invention is to develop a drug delivery system capable of suitably delivering a drug to the heart and / or the diaphragm.SOLUTION: According to the present invention, there are provided a particle for drug delivery to the heart and / or diaphragm having a boronic acid group as a targeting moiety, and a pharmaceutical composition and the like comprising the particle for drug delivery.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to particles for drug delivery to the heart and / or diaphragm and pharmaceutical compositions containing the particles for drug delivery. [Background technology]

[0002] Various drug delivery systems (DDS) have been developed to efficiently deliver drugs. For example, liposome formulations and micelle formulations containing drugs have been investigated (Patent Documents 1 and 2, etc.).

[0003] In the development of the above-mentioned DDS, in order to achieve highly target-selective drug delivery, targeting moieties (e.g., ligands) that can specifically recognize targets are introduced into drugs or drug transporters to impart targeting properties to them.

[0004] On the other hand, the heart and diaphragm are among the organs with low drug delivery efficiency, and a DDS that can efficiently deliver drugs to these organs is needed. For example, in the treatment of diseases such as Duchenne muscular dystrophy and heart failure, a DDS that can efficiently deliver drugs to the heart is needed. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] WO2019 / 240223 [Patent Document 2] WO2013 / 162041 Summary of the Invention [Problem to be solved by the invention]

[0006] The main object of the present invention is to develop a DDS that can efficiently deliver drugs to the heart and / or diaphragm. [Means for solving the problem]

[0007] The present inventors have found that a boronic acid group can function as a targeting moiety in drug delivery to the heart and / or diaphragm, and have completed the present invention. [1] According to one aspect of the present invention, there is provided a particle for drug delivery to the heart and / or diaphragm, which has a boronic acid group as a targeting moiety. [2] In the drug delivery particle according to [1] above, the boronic acid group may include a phenylboronic acid group or a pyridineboronic acid group. [3] The particle diameter of the drug delivery particles described in [1] or [2] above may be 100 nm or less. [4] The drug delivery particle according to any one of [1] to [3] above may comprise a drug transporter component molecule to which the boronic acid group and a drug are bound. [5] In the drug delivery particle according to [4] above, the drug transporter component molecule may be a linear polymer, a branched polymer, or a graft polymer. [6] The drug delivery particle described in any one of [1] to [3] above may each contain a specific number of complexes of a drug and a drug transporter component molecule, and the boronic acid group may be bound to at least one of the drug transporter component molecule and the drug. [7] The drug delivery particle according to any one of [1] to [3] above may comprise a micelle or vesicle containing the drug transporter component molecule having the boronic acid group bound thereto. [8] The drug delivery particle described in [7] above may further contain a drug, and the drug may be encapsulated in the micelle or the vesicle. [9] The drug delivery particle according to any one of [1] to [3] above may contain a drug having the boronic acid group bound thereto.

[10] The drug delivery particles according to any one of [1] to [9] above may be used for the treatment of diseases requiring delivery of a drug to the heart and / or diaphragm.

[11] According to another aspect of the present invention, there is provided a pharmaceutical composition comprising the drug delivery particle according to any one of [1] to

[10] above.

[12] According to another aspect of the present invention, there is provided the use of a boronic acid group to direct a drug or drug transporter to the heart and / or diaphragm.

[13] According to another aspect of the present invention, there is provided use of a boronic acid compound for producing a drug delivery particle according to any one of [1] to

[10] above. [Effects of the Invention]

[0008] According to the drug delivery particles of the embodiments of the present invention, the boronic acid group exerts directionality toward the heart and / or diaphragm, thereby enabling efficient drug delivery to the heart and / or diaphragm. Although this effect is not intended to limit the present invention, it is presumed that the boronic acid group functions as a ligand that specifically recognizes the cell surface of the heart (specifically, the surfaces of cardiomyocytes and other cells that constitute the heart). [Brief explanation of the drawings]

[0009] [Figure 1] FIG. 1 is a schematic diagram illustrating an example of the structure of a micelle according to one embodiment of the present invention. [Figure 2] FIG. 1 is a schematic diagram illustrating an example of the structure of a liposome according to one embodiment of the present invention. [Figure 3A] FIG. 1 is a schematic diagram illustrating an example of the structure of a polyion complex according to one embodiment of the present invention. [Figure 3B] FIG. 1 is a schematic diagram illustrating an example of the structure of a polyion complex according to one embodiment of the present invention. [Figure 4] FIG. 1 shows the biodistribution of drug delivery particles composed of a hyperbranched polymer to which boronic acid groups are attached. [Figure 5] 5A and 5B show the biodistribution of drug delivery particles composed of grafted polymers to which boronic acid groups are attached. DETAILED DESCRIPTION OF THE INVENTION

[0010] Preferred embodiments of the present invention will be described below, but the present invention is not limited to these embodiments. The embodiments can be combined as appropriate unless such understanding is clearly inappropriate from the context. Furthermore, in this specification, the term "to" indicating a range of values ​​includes both the upper and lower limits.

[0011] A. Drug Delivery Particles The drug delivery particle according to an embodiment of the present invention is a drug delivery particle used for delivering a drug to the heart and / or diaphragm. The drug delivery particle has a boronic acid group (-B(OH)2) as a targeting moiety that is directed to the heart and / or diaphragm.

[0012] The drug delivery particles are typically nanoparticles having a particle diameter of less than 1 μm, preferably 100 nm or less. The particle diameter of the drug delivery particles is, for example, 5 nm to 50 nm, and may be 6 nm or more, 8 nm or more, or 40 nm or less, 35 nm or less, or 30 nm or less. When the particle diameter of the drug delivery particles is within the above range, excretion from the kidney into urine and nonspecific accumulation in organs other than the target can be suppressed. The particle diameter is a hydrodynamic diameter and can be determined, for example, by dynamic light scattering (DLS) or fluorescence correlation spectroscopy (FCS). In this specification, the drug delivery particles may be any object having the above particle diameter as a hydrodynamic diameter. Therefore, the drug delivery particles are not limited to spherical shapes and may have any shape. Furthermore, the drug delivery particles may be composed of a single molecule or an aggregate of multiple molecules.

[0013] Drug delivery particles may or may not contain a drug. In one embodiment, drug delivery particles contain a drug to which a boronic acid group has been introduced (bound). In another embodiment, drug delivery particles contain a drug transporter (DDS carrier) component molecule to which a boronic acid group has been introduced (bound). The DDS carrier component molecules may constitute a DDS carrier alone, or in combination with a drug, when multiple DDS carrier component molecules are combined to constitute a DDS carrier, a single DDS carrier component molecule may be used alone, or two or more may be used in combination. Furthermore, the DDS carrier component molecule may be bound to a drug.

[0014] The boronic acid group can be introduced into drug delivery particles in any suitable form as long as the effects of the present invention are achieved. The drug delivery particles may have, for example, an aliphatic boronic acid group in which -B(OH)2 is introduced into an aliphatic hydrocarbon group and / or an aromatic boronic acid group in which -B(OH)2 is introduced into an aromatic ring. Examples of the aliphatic hydrocarbon group include linear or branched alkyl groups, alkenyl groups, and alkynyl groups, which may contain a cyclic structure. The aliphatic hydrocarbon group may have one or more carbon atoms, and may have, for example, 24 or less, 12 or less, or 6 or less. The aromatic ring may be an aromatic hydrocarbon ring whose ring-constituting atoms are solely carbon atoms, or an aromatic heterocycle containing a heteroatom as a ring-constituting atom. The aromatic ring may be a monocyclic aromatic ring or a polycyclic aromatic ring.

[0015] The aliphatic boronic acid group can be, for example, a group represented by the following formula (1): -R 1 -B(OH)2(1) (In the formula, R 1 is a divalent group formed by removing two hydrogens from a straight-chain or branched alkane, alkene, or alkyne, which may contain a cyclic structure. It can be expressed as:

[0016] The aromatic boronic acid group is, for example, represented by the following formula (2): -Ar-B(OH)2(2) (In the formula, Ar is an aromatic ring.) It can be expressed as:

[0017] Regarding formula (2), specific examples of the aromatic ring include a benzene ring, a pyridine ring, a naphthalene ring, a biphenyl ring, etc. Among these, the aromatic boronic acid group is preferably a phenylboronic acid group in which Ar is a benzene ring or a pyridineboronic acid group in which Ar is a pyridine ring.

[0018] In the benzene ring and the pyridine ring, the carbon atom to which -B(OH)2 is bonded (the carbon atom having a free valence is numbered as 1) may be 2, 3, or 4, for example, 3 or 4, and preferably 4. In the pyridine boronic acid group, the nitrogen atom in the pyridine ring may be 2, 3, or 4, for example, 2 or 3, and preferably 2.

[0019] The aromatic ring may have a substituent at any position on the ring. Examples of the substituent include halogens such as fluorine, chlorine, and bromine, alkyl groups having 1 to 3 carbon atoms (e.g., methyl groups), fluorinated alkyl groups having 1 to 3 carbon atoms (e.g., trifluoromethyl groups), alkoxy groups having 1 to 3 carbon atoms (e.g., methoxy groups), formyl groups, nitro groups, and amino groups. The aliphatic hydrocarbon groups may also have a substituent at any position. Examples of the substituent include halogens such as fluorine, chlorine, and bromine, nitro groups, and amino groups. The number of substituents in the aromatic ring or the aliphatic hydrocarbon group may be 0 to 4, for example, 0 to 2.

[0020] The phenylboronic acid group can be, for example, a group represented by the following formula (3): [ka] (In formula (3), n1 is an integer of 0 to 4, for example, 0, 1, or 2, and when n1 is 1, the introduction site of F and B(OH)2 may be any of ortho, meta, and para.)

[0021] The pyridine boronic acid group is, for example, a group represented by the following formula (4): [ka] (In formula (4), n2 is an integer of 0 to 3, for example, 0, 1, or 2, and when n2 is 1, the introduction site of F and B(OH)2 may be any of ortho, meta, and para.)

[0022] A-1. Drug delivery particles containing DDS carrier molecules to which boronic acid groups and drugs are bound The DDS carrier component molecule to which a boronic acid group and a drug are bound is a conjugate between the DDS carrier component molecule to which a boronic acid group is bound and the drug. Drug delivery particles containing a drug transporter component to which a boronic acid group and a drug are bound can be composed of the conjugate. The number of DDS carrier components contained in the drug delivery particle can be one.

[0023] Biocompatible polymers are preferably used as DDS carrier constituent molecules. The biocompatible polymer is preferably a hydrophilic polymer. Examples of the hydrophilic polymer include poly(ethylene glycol), poly(saccharide), poly(vinylpyrrolidone), poly(vinyl alcohol), poly(acrylamide), poly(acrylic acid), poly(methacrylamide), poly(methacrylic acid), poly(methacrylic acid ester), poly(acrylic acid ester), poly(amino acid) (e.g., poly(aspartic acid), poly(glutamic acid), poly(serine)), poly(malic acid), and copolymers or derivatives thereof. Among these, poly(ethylene glycol) is preferably used. Various poly(ethylene glycol)s with terminal functional groups are commercially available.

[0024] The polymer may be a linear polymer, a branched polymer (including a dendrimer or a dendron), or a graft polymer. By changing the chain length of the linear polymer, the number or length of branches of the branched polymer, or the number or length of graft chains of the graft polymer, a conjugate with a desired particle size can be suitably obtained.

[0025] The molecular weight (Mw) of the linear polymer may be, for example, 100 to 1,000,000, preferably 500 to 100,000, more preferably 1,000 to 50,000, and may be, for example, 3,000 or more, 5,000 or more, or 10,000 or more. The molecular weight of the polymer can be determined, for example, by size exclusion chromatography.

[0026] The number of polymer chains in the branched polymer may be, for example, 2 or more, 5 or more, or 6 or more, and may be, for example, 200 or less, or 50 or less.

[0027] The molecular weight (Mw) of each polymer chain of the branched polymer can be independently, for example, 100 to 1,000,000, preferably 500 to 100,000, and more preferably 1,000 to 50,000. In one embodiment, each polymer chain of the branched polymer can have the same molecular weight.

[0028] The molecular weight (Mw) of the branched polymer as a whole may be, for example, 200 to 2,000,000, preferably 1,000 to 200,000, more preferably 2,000 to 100,000, and may be, for example, 3,000 or more, 5,000 or more, or 10,000 or more.

[0029] The boronic acid group and the drug may be attached to any position on the linear polymer and the branched polymer, respectively. The boronic acid group may be attached preferably to the end of at least one polymer chain. The drug may be attached to any position on the polymer chain, or may be attached to the end of the polymer chain. The number of boronic acid groups attached to one molecule of the linear polymer and the branched polymer, respectively, may be, for example, 1 to 200, or 1 to 100. The number of drugs attached to one molecule of the linear polymer and the branched polymer, respectively, may be, for example, 1 to 200, or 1 to 50, or 1 to 10.

[0030] The graft polymer may be a polymer in which a biocompatible polymer is grafted to the side chain of a trunk polymer. The biocompatible polymer is preferably a hydrophilic polymer, specific examples of which are as described above.

[0031] Examples of polymers that make up the trunk polymer include poly(amino acids) such as poly(aspartic acid), poly(glutamic acid), poly(aspartic acid)-poly(glutamic acid) copolymer, and poly(lysine), poly(acrylic acid), poly(vinyl alcohol), chitosan, and poly(vinylamine).

[0032] The number of graft chains in the graft polymer is, for example, 1 or more, preferably 5 or more, and may be 20 or more or 50 or more, and may be, for example, 500 or less, or 100 or less.

[0033] The molecular weight (Mw) of each graft chain of the graft polymer can be independently, for example, 500 to 100000, preferably 500 to 50000, and more preferably 1000 to 20000. In one embodiment, each graft chain can have the same molecular weight.

[0034] The number of structural units of the trunk polymer (degree of polymerization) can be, for example, 5 to 1000, and preferably 10 to 200. The introduction rate of graft chains (number of introduced graft chains / degree of polymerization of trunk polymer × 100) can be, for example, 1% to 100%, 5% to 80%, or 10% to 80%.

[0035] The molecular weight (Mw) of the graft polymer as a whole may be, for example, 500 to 1,000,000, preferably 10,000 to 500,000, and more preferably 20,000 to 200,000.

[0036] The boronic acid group and the drug may each be bound to any position on the graft polymer. The boronic acid group may preferably be bound to the end of the graft chain. The drug may be bound to any position on the graft polymer, and may be bound to the end of the main chain or the end of the graft chain. The number of boronic acid groups bound to one molecule of the graft polymer is, for example, 1 to 1000, 1 to 100, or 1 to 50. The number of drugs bound to one molecule of the graft polymer is, for example, 1 to 1000, 1 to 500, or 1 to 100.

[0037] Any appropriate substance can be used as the drug depending on the purpose. The drug can be, for example, a substance having various physiological activities such as cytostatic activity, antitumor activity, immunomodulatory activity, antiviral activity, antibacterial activity, or anti-inflammatory activity. The drug can also be a detection reagent such as a fluorescent dye or a contrast agent, or a cell.

[0038] Specific examples of physiologically active drugs include proteins (e.g., antibodies or functional fragments thereof, enzymes, hormones), nucleic acids (e.g., high molecular weight nucleic acids such as plasmid DNA and mRNA, and low molecular weight nucleic acids such as siRNA, miRNA, antisense nucleic acids, and aptamers), and other physiologically active substances (e.g., antitumor agents, signal transduction inhibitors, metabolic antagonists, analgesics, anti-inflammatory agents, and antibacterial agents).

[0039] The drug may be a low molecular weight drug (e.g., less than 1 kDa) or a high molecular weight drug, e.g., 1 kDa or more, 5 kDa or more, or 10 kDa or more, e.g., 1000 kDa or less, 200 kDa or less, or 50 kDa or less.

[0040] The binding of a drug to a DDS carrier component molecule can be carried out by any suitable method. For example, a drug having a functional group A and a DDS carrier component molecule having a functional group B capable of binding to functional group A can be bound to the DDS carrier component molecule by reacting functional group A with functional group B. In this case, the functional group A of the drug may be inherent in the drug or may be bound to the drug via a linker.

[0041] Similarly, the introduction of a boronic acid group into a DDS carrier constituent molecule can be carried out by binding a boronic acid compound having a boronic acid group and another functional group to the DDS carrier constituent molecule directly or via a linker.

[0042] A-2. Drug delivery particles containing drug transporter molecules bound to boronic acid groups Drug delivery particles containing DDS carrier constituent molecules bound to boronic acid groups can be, for example, micelles or vesicles containing DDS carrier constituent molecules bound to boronic acid groups, polyion complexes (PICs) containing DDS carrier constituent molecules bound to boronic acid groups and drugs, hydrogel particles composed of DDS carrier constituent molecules bound to boronic acid groups, etc. Examples of drugs include those described in Section A-1.

[0043] A-2-1. Micelle A micelle can be a particulate aggregate formed by the aggregation of amphiphilic molecules having a hydrophilic segment and a hydrophobic segment. A micelle may encapsulate a drug inside. The drug encapsulated in the micelle may or may not be bound to a DDS carrier component molecule.

[0044] Examples of micelles include those formed by arranging block copolymers having a first polymer segment and a second polymer segment having a lower hydrophilicity than the first polymer segment in a generally radial pattern with the second polymer segment facing inward. As shown in Figure 1, micelle 100 includes, as DDS carrier constituent molecules, block copolymer 110a having first polymer segment 112, second polymer segment 114, and boronic acid group 116 attached to the end of first polymer segment 112. Optionally, micelle 100 may further include block copolymer 110b having first polymer segment 112 and second polymer segment 114, but no boronic acid group attached.

[0045] The number of boronic acid groups introduced per micelle is not limited as long as the effects of the present invention are obtained, and can be appropriately selected depending on the size of the micelle, etc. The number of boronic acid groups introduced is, for example, 1 to 10,000, and may be 5 or more, or 10 or more, or 1,000 or less, 100 or less, or 20 or less.

[0046] The first polymer segment typically includes a biocompatible hydrophilic polymer. Examples of biocompatible hydrophilic polymers include those described in Section A-1. Among these, polymers that are uncharged at physiological pH (pH 7.4) are preferred, nonionic polymers are more preferred, and poly(ethylene glycol) is even more preferred.

[0047] The molecular weight (Mw) of the hydrophilic polymer may be, for example, 1,000 to 100,000, preferably 2,000 to 60,000, more preferably 5,000 to 50,000, and even more preferably 10,000 to 40,000.

[0048] In the illustrated example, the first polymer segment is composed of a linear polymer, but the first polymer segment may also be composed of a branched polymer or a graft polymer. Examples of branched polymers and graft polymers include those described in Section A-1. In the illustrated example, one boronic acid group is bonded to each block copolymer molecule, but two or more boronic acid groups may be bonded to each block copolymer molecule. The boronic acid group may be bonded, for example, to the main chain or branched chain end, or the side chain or graft chain end of the first polymer segment of the block copolymer. The boronic acid group is preferably located on the outer surface of the micelle. In other words, the micelle preferably contains boronic acid groups exposed on its outer surface.

[0049] The second polymer segment may comprise, for example, a poly(amino acid). Appropriate selection of the side chains of the poly(amino acid) allows for adjustment of micelle stability, drug retention, and other properties. For example, hydrophobic side chains in the poly(amino acid) may result in hydrophobic interactions between block copolymers and / or between the block copolymer and a hydrophobic drug. Alternatively, electrostatic interactions between the block copolymer and an anionic or cationic drug may occur when the poly(amino acid) has side chains containing cationic or anionic groups. Alternatively, drugs may be attached to the side chains of the second polymer segment.

[0050] The degree of polymerization of the second polymer segment may be, for example, 10-200, preferably 15-150, and more preferably 20-100.

[0051] The block copolymer to which the boronic acid group is attached can be, for example, a block copolymer represented by the formula: ZL 1 -Xa-L 2 -Xb (wherein Z represents a boronic acid group, Xa represents a first polymer segment, Xb represents a second polymer segment, and L 1 and L 2 each independently represents a single bond or a linking group.

[0052] L 1 and L 2 The divalent linking group that can be represented by is not particularly limited as long as the effects of the present invention can be obtained. The linking group can be, for example, a linear or branched alkylene group having 1 to 6 carbon atoms, -COO-, -CONH-, -NH-, -CO-, -O-, -S-, or any combination thereof. The linking group may also contain a ring structure formed by click chemistry (e.g., a 1,2,3-triazole ring). The length of the linking group (the number of atoms connecting the shortest distance between the boronic acid group to which the linking group is bonded and the drug residue) can be, for example, 1 to 30, 1 to 20, or 1 to 10.

[0053] For specific examples of the block copolymer, reference can be made to WO2007 / 099660, WO2007 / 099661, WO2010 / 093036, WO2015 / 170757, etc.

[0054] Various methods are known for forming the micelles and encapsulating drugs therein, and those skilled in the art can select any appropriate method.

[0055] A-2-2. Vesicles The vesicles may be closed sac-like aggregates formed by assembling a plurality of DDS carrier constituent molecules, including DDS carrier constituent molecules bound to boronic acid groups. The vesicles may encapsulate a drug inside. The drug may or may not be bound to the DDS carrier constituent molecules.

[0056] The number of boronic acid groups introduced per vesicle is not limited as long as the effects of the present invention are obtained, and can be appropriately selected depending on the size of the vesicle, etc. The number of boronic acid groups introduced is, for example, 1 to 10,000, and can be 5 or more or 10 or more, or 1,000 or less, 100 or less, or 20 or less. The boronic acid groups are preferably located on the outer surface of the vesicle. In other words, the vesicle preferably contains boronic acid groups exposed on its outer surface.

[0057] Vesicles include liposomes and polymersomes.

[0058] Liposomes may be vesicles formed, for example, by a two-layer molecular membrane. The two-layer molecular membrane can be formed by arranging amphipathic lipid molecules having a hydrophobic portion and a hydrophilic portion in a double layer with the hydrophobic portion facing inward. As shown in FIG. 2 , liposome 200 includes, as a DDS carrier component molecule, amphipathic lipid molecule 210a having a hydrophobic portion 212, a hydrophilic portion 214, and a boronic acid group 216 bound to the hydrophilic portion 214. If necessary, liposome 200 may further include amphipathic lipid molecule 210b having a hydrophobic portion 212 and a hydrophilic portion 214 but no boronic acid group bound to the amphipathic lipid molecule. In the illustrated example, one boronic acid group is bound to each amphipathic lipid molecule; however, two or more boronic acid groups may be bound to each amphipathic lipid molecule.

[0059] The amphipathic lipid molecule may be any biocompatible molecule, and a glycerophospholipid such as phosphatidylcholine may be preferably used. The boronic acid group may be bonded to the end of the hydrophilic portion of the amphipathic lipid molecule.

[0060] The liposome may be surface-modified with a biocompatible hydrophilic polymer. In this case, the hydrophilic polymer is introduced into the hydrophilic portion of the amphiphilic lipid molecule, and a boronic acid group may be introduced at the end of the hydrophilic polymer. Examples of biocompatible hydrophilic polymers include those described in Section A-1. Among them, poly(ethylene glycol) is preferred.

[0061] The amphiphilic lipid molecule having a boronic acid group attached thereto may have a structure represented by the formula Z-Xa-Xb or Z-A-Xa-Xb, where Z represents the boronic acid group, Xa represents the hydrophilic portion, Xb represents the hydrophobic portion, and A represents the hydrophilic polymer segment.

[0062] The vesicle may be a lipid nanoparticle. The lipid nanoparticle may have a structure in which an ionized lipid molecule and a drug (e.g., nucleic acid) are encapsulated in a lipid membrane containing so-called ionized lipid molecules, PEGylated lipid molecules, helper phospholipid molecules, and cholesterol. The lipid nanoparticle contains at least one lipid molecule having a boronic acid group attached thereto as a DDS carrier component. The boronic acid group may be attached, for example, to the PEG chain end of the PEGylated lipid molecule.

[0063] Examples of polymersomes include polymersomes composed of a cationic block copolymer having an uncharged hydrophilic polymer segment and a cationic polymer segment, and an anionic block copolymer or anionic homopolymer having an uncharged hydrophilic polymer segment and an anionic polymer segment, or polymersomes composed of the anionic block copolymer and the cationic block copolymer or cationic homopolymer. Such polymersomes can be formed by electrostatically complexing the cationic polymer segment or cationic homopolymer with the anionic polymer segment or anionic homopolymer, resulting in a spherical assembly with the uncharged hydrophilic polymer segment on the outside. Note that the cationic homopolymer refers to a polymer composed solely of cationic polymer segments, without both an uncharged hydrophilic polymer segment and a cationic polymer segment. Similarly, the anionic homopolymer refers to a polymer composed solely of anionic polymer segments, without both an uncharged hydrophilic polymer segment and an anionic polymer segment. Thus, each of these homopolymers may contain two or more types of structural units.

[0064] The uncharged hydrophilic polymer constituting the uncharged hydrophilic polymer segment may be poly(ethylene glycol). The cationic polymer or the cationic polymer constituting the cationic polymer segment may be a cationic poly(amino acid). The anionic polymer or the anionic polymer constituting the anionic polymer segment may be an anionic poly(amino acid). Specific examples of the polymersome and its constituent polymers may be found in WO2012 / 014942, etc.

[0065] The polymersome may contain, as a DDS carrier component molecule, a cationic block copolymer having a boronic acid group attached to the end of an uncharged hydrophilic polymer segment, and / or an anionic block copolymer having a boronic acid group attached to the end of an uncharged hydrophilic polymer segment. By using such a cationic block copolymer and / or anionic block copolymer, a polymersome having boronic acid groups arranged on its surface can be obtained.

[0066] Various methods are known for forming the above-mentioned various vesicles and for encapsulating drugs therein, and those skilled in the art can select any appropriate method.

[0067] A-2-3. Polyion complex (PIC) Examples of PICs include PICs between cationic polymers and anionic drugs, and PICs between anionic polymers and cationic drugs. Examples of anionic drugs include nucleic acids and proteins. Examples of cationic drugs include proteins.

[0068] PICs of cationic polymers and anionic drugs include complexes (unit PICs (uPICs)) consisting of structural units containing specific numbers of cationic block copolymers having hydrophilic and cationic polymer segments, and specific numbers of anionic drugs. For example, a uPIC contains one anionic drug and one or more cationic block copolymers. The uPIC may be, for example, a uPIC composed of one molecule of cationic block copolymer and one molecule of anionic drug, a uPIC composed of two molecules of cationic block copolymer and one molecule of anionic drug, or a uPIC composed of three molecules of cationic block copolymer and one molecule of anionic drug. The boronic acid group may be bound to either the cationic block copolymer or the anionic drug. In the former case, the boronic acid group may be bound to the hydrophilic polymer segment of the cationic block copolymer. In the latter case, the boronic acid group may be bound to the drug via the hydrophilic polymer. Alternatively, the boronic acid group may be bound to both the cationic block copolymer and the anionic drug, which may be a nucleic acid having a base length (base pair length in the case of a double-stranded nucleic acid) of 10 to 50 or 15 to 30.

[0069] The hydrophilic polymer segment of the cationic block copolymer typically contains a biocompatible hydrophilic polymer. The hydrophilic polymer may be a linear polymer, a branched polymer, or a graft polymer. Examples of such hydrophilic polymers include those described in Section A-1.

[0070] The molecular weight (Mw) of the hydrophilic polymer may be, for example, 1,000 to 100,000, preferably 10,000 to 50,000, and more preferably 20,000 to 40,000. When the hydrophilic polymer is a branched polymer (for example, a bibranched polymer) or a graft polymer, the molecular weight of the hydrophilic polymer may be the molecular weight of each branched chain or grafted chain.

[0071] The cationic polymer segment of the cationic block copolymer includes, for example, a cationic poly(amino acid). The cationic poly(amino acid) includes, for example, cationic amino acid residues having a cationic group (typically, an amino group) in the side chain. Examples of the cationic amino acid residue include basic amino acid residues such as lysine residue, ornithine residue, arginine residue, and histidine residue. Another example of the cationic amino acid residue is an aspartic acid residue or glutamic acid residue in which the -OH moiety of the carboxyl group (-C(O)OH) in the side chain is -NH-(CH2). p1 -〔NH-(CH2) q1 -] r1 Examples include amino acid residues substituted with an NH group (wherein p and q are each independently an integer of 1 to 5, preferably 2 or 3, and more preferably 2, and r is an integer of 1 to 5, preferably an integer of 1 to 3).

[0072] Figure 3A is a schematic diagram illustrating an example of a uPIC composed of two molecules of a cationic block copolymer to which a boronic acid group is attached and one molecule of an anionic drug. uPIC300A is composed of two molecules of a cationic block copolymer 310 and one molecule of a nucleic acid 320. Cationic block copolymer 310 has a hydrophilic polymer segment 312 and a cationic polymer segment 314, and has a boronic acid group 316 attached to the end of hydrophilic polymer segment 312. uPIC300A is formed by electrostatic interaction between cationic polymer segment 314 of cationic block copolymer 310 and anionic drug 320.

[0073] For specific examples of the above uPIC, reference can be made to WO2013 / 162041, WO2019 / 044937, etc.

[0074] In the illustrated example, the hydrophilic polymer segment 312 is composed of a linear polymer, but the hydrophilic polymer segment may also be composed of a branched polymer or a graft polymer. Examples of branched polymers and graft polymers include those described in Section A-1. In the illustrated example, one boronic acid group is bonded to each cationic block copolymer molecule, but two or more boronic acid groups may be bonded to each cationic block copolymer molecule. The boronic acid group may be bonded, for example, to the main chain or branched chain end, or the side chain or graft chain end of the hydrophilic polymer segment of the cationic block copolymer. Furthermore, unlike the illustrated example, the uPIC may be composed of one or more molecules of cationic block copolymer 310 and one molecule of nucleic acid 320. In this embodiment, it is sufficient that a boronic acid group is bonded to at least one of the cationic block copolymers constituting the uPIC. Thus, for example, the uPIC may be composed of a cationic block copolymer having a boronic acid group, a cationic block copolymer not having a boronic acid group, and an anionic drug.

[0075] FIG. 3B is a schematic diagram illustrating an example of a uPIC composed of one molecule of a cationic block copolymer and one molecule of an anionic drug to which a boronic acid group is bound via a hydrophilic polymer. Cationic block copolymer 330 has a hydrophilic polymer segment 332 and a cationic polymer segment 334. Anionic drug 320 is bound to boronic acid group 344 via hydrophilic polymer segment 342. uPIC 300B is formed by electrostatic interaction between cationic polymer segment 334 of cationic block copolymer 330 and anionic drug 320. In this embodiment, conjugate 340 of anionic drug 320, hydrophilic polymer segment 342, and boronic acid group 344, and cationic block copolymer 330 together function as DDS carrier constituent molecules.

[0076] In the illustrated example, the cationic block copolymer 330 does not have a boronic acid group, but a boronic acid group may be bonded to the cationic block copolymer 330 (preferably the hydrophilic polymer segment 332). Also, while the cationic block copolymer 330 contained in the illustrated uPIC300B is one molecule, it may contain two or more molecules. Furthermore, in the illustrated example, the hydrophilic polymer segments 332 and 342 are composed of linear polymers, but each of these hydrophilic polymer segments may be composed of a branched polymer or a graft polymer. Examples of branched polymers and graft polymers include those described in Section A-1. In the illustrated example, one boronic acid group is bonded to one molecule of the conjugate, but two or more boronic acid groups may be bonded to one molecule of the conjugate. The boronic acid group may be bonded, for example, to the main chain or branched chain end, or the side chain or graft chain end of the hydrophilic polymer segment of the conjugate.

[0077] Other examples of PICs include PICs between cationic poly(amino acids) and anionic drugs, or PICs between anionic poly(amino acids) and cationic drugs. The cationic poly(amino acids) contain cationic amino acid residues with cationic groups in their side chains. The anionic poly(amino acids) contain anionic amino acid residues with anionic groups in their side chains. The PICs may be composed of an unspecified number of poly(amino acids) and an unspecified number of drug molecules. Such PICs may contain poly(amino acids) with boronic acid groups as DDS carrier constituent molecules, and may further contain poly(amino acids) without boronic acid groups, depending on the purpose. The boronic acid group may be bound, for example, to at least one end of the poly(amino acids). If necessary, any appropriate linker or hydrophilic polymer segment may be interposed between the poly(amino acids) and the boronic acid group.

[0078] A-3. Drug delivery particles containing a drug having a boronic acid group bound thereto The drug delivery particle containing a drug having a boronic acid group bonded thereto may be composed of the above-mentioned drug having a boronic acid group bonded thereto. The number of drugs contained in the drug delivery particle containing a drug having a boronic acid group bonded thereto may be 1.

[0079] Examples of drugs include those described in section A-1.

[0080] The number of boronic acid groups bonded per drug molecule is not limited as long as the effects of the present invention are obtained, and can be appropriately selected depending on the drug's chemical structure, steric structure, molecular weight, etc. The number of boronic acid groups bonded is, for example, 1 to 200, and may be 3 or more or 5 or more, or 100 or less, 50 or less, or 10 or less.

[0081] The boronic acid group may be directly bound to the drug or may be bound via a linker. In one embodiment, the boronic acid group is bound to one drug via two or more linkers.

[0082] The linker is not particularly limited, and examples thereof include the linkers described in Section A-2. Furthermore, for example, a biocompatible polymer can be used as the linker. The biocompatible polymer is preferably a hydrophilic polymer. Furthermore, the biocompatible polymer may be a linear polymer, a branched polymer, or a graft polymer. Examples of biocompatible hydrophilic polymers include those described in Section A-1. The boronic acid group can be bonded, for example, to the main chain or branched chain end, or the side chain or graft chain end of the polymer. A specific example of an embodiment using a branched polymer as a linker is an embodiment in which a drug is bonded to the central portion of a dendron having a boronic acid group introduced into one or more of its terminal groups. In this embodiment, the number of dendrons bonded to a drug may be two or more.

[0083] B. Pharmaceutical Compositions Pharmaceutical compositions according to embodiments of the present invention contain the drug delivery particles described in Section A and may further contain any appropriate additives depending on the intended purpose. The type and amount of additives can be determined appropriately by those skilled in the art depending on the intended purpose. Specific examples of additives include excipients, tonicity adjusting agents, pH adjusters, buffers, stabilizers, etc.

[0084] In one embodiment, the pharmaceutical composition is an injectable preparation. The injectable pharmaceutical composition may be a powder formulation, a lyophilized formulation, or a liquid formulation. The liquid formulation, for example, contains an aqueous medium, which may be buffered if necessary. When the drug delivery particles are micelles, vesicles, or PICs, the micelles, vesicles, or PICs may be formed in the aqueous medium. On the other hand, the powder formulation and the lyophilized formulation are dissolved or dispersed in an aqueous medium such as water for injection before use. When the drug delivery particles are micelles, vesicles, or PICs, the micelles, vesicles, or PICs may be formed in the solution or dispersion.

[0085] The disease to be treated by the pharmaceutical composition may be a disease requiring delivery of a drug to the heart and / or diaphragm. Such diseases include Duchenne muscular dystrophy, heart failure, myocardial infarction, dilated cardiomyopathy, cardiac hypertrophy, etc. For example, the treatment of Duchenne muscular dystrophy using antisense nucleic acids has been investigated.

[0086] The pharmaceutical composition may be administered via intravenous administration, intracoronary administration, intramyocardial administration, etc. The subject of administration is typically a human or a non-human mammal (such as a mouse, rat, rabbit, monkey, dog, or horse). [Example]

[0087] The present invention will be specifically explained below with reference to examples, but the present invention is not limited to these examples in any way.

[0088] [Experimental Example 1: Drug delivery particles containing 8-branched poly(ethylene glycol) as a DDS carrier component] 1. Raw materials Amine-terminated 8-arm polyethylene glycol (8arm-PEG), molecular weight 20,000 (purchased from SINOPEG, used as is, product name: 8armPEG-NH2(TP)) N,N-dimethylformamide (DMF) (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) Dichloromethane (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) Acetic anhydride (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) Methanol (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) Succinic anhydride (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) Triethylamine (TEA) (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) Sodium bicarbonate (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) 4-(4,6-Dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-MM) (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) 4-Carboxy-3-fluorophenylboronic acid (FPBA) (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) p-Carboxyphenylboronic acid (PBA) (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) m-Aminophenylboronic acid (APBA) (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) Benzoyl chloride (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) D-PBS(-) (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) Sulfo-Cy5-NHS (purchased from Lumiprobe, dissolved in DMSO at a concentration of 25 mg / mL and used as is) 6-Carboxypyridine-3-boronic acid (PyBA) (purchased from Combi-Block, used as is)

[0089] 2-1.Synthesis of Cy5-8arm-PEG 500 mg of 8-arm PEG was dissolved in 10 mL of DMF, and 15.5 mg of sulfo-Cy5-NHS (622 μL) was added. The mixture was stirred overnight and the reaction mixture was placed in a dialysis membrane with a molecular weight cutoff of 3500 and dialyzed for 2 days. The mixture was then recovered by lyophilization.

[0090] 2-2. Synthesis of Cy5-8arm-PyBA (Introduction of PyBA) PyBA (17 mg) and sodium bicarbonate (8 mg) were added to Cy5-8arm-PEG (50 mg), and 2 mL of DMF was added and stirred. DMT-MM (30 mg) was then added and allowed to react overnight. The reaction solution was filtered through a 0.45 μm filter. The filtrate was placed in a dialysis membrane with a molecular weight cutoff of 3500 and dialyzed for two days, after which it was recovered by lyophilization.

[0091] 2-3. Synthesis of Cy5-8arm-FPBA (Introduction of FPBA) FPBA (19 mg) was added to Cy5-8arm-PEG (50 mg) and dissolved in 2 mL of methanol. DMT-MM (30 mg) was then added and reacted overnight. The reaction solution was placed in a dialysis membrane with a molecular weight cutoff of 3500 and dialyzed for two days, after which the solution was recovered by lyophilization.

[0092] 2-4. Synthesis of Cy5-8arm-PBA (Introduction of PBA) PBA (17 mg) was added to Cy5-8arm-PEG (50 mg) and dissolved in 2 mL of methanol. DMT-MM (30 mg) was then added and allowed to react overnight. The reaction solution was placed in a dialysis membrane with a molecular weight cutoff of 3500 and dialyzed for 2 days, after which the solution was recovered by lyophilization.

[0093] 2-5. Synthesis of Cy5-8arm-APBA (Introduction of APBA) Cy5-8arm-PEG (50 mg) was dissolved in 2 mL of dichloromethane, and TEA (6 μL) and succinic anhydride (4 mg) were added and reacted overnight. The reaction solution was placed in a dialysis membrane with a molecular weight cutoff of 3500 and dialyzed for 2 days, after which the product was recovered by lyophilization. The recovered Cy5-8arm-PEG-succinic acid was then dissolved in 2 mL of methanol, and APBA (14 mg), DMT-MM (30 mg), and sodium bicarbonate (4 mg) were added and reacted overnight. The reaction solution was placed in a dialysis membrane with a molecular weight cutoff of 3500 and dialyzed for 2 days, after which the product was recovered by lyophilization.

[0094] 2-6. Synthesis of Cy5-8arm-Phenyl (Introduction of Phenyl) Cy5-8arm-PEG (50 mg) was dissolved in 2 mL of dichloromethane, and TEA (14 μL) and benzoyl chloride (12 μL) were added and reacted overnight. The reaction solution was placed in a dialysis membrane with a molecular weight cutoff of 3500 and dialyzed for 2 days, after which the product was recovered by lyophilization.

[0095] 2-7. Synthesis of Cy5-8arm-Ace (Introduction of Ace) Cy5-8arm-PEG (50 mg) was dissolved in 2 mL of dichloromethane, and TEA (14 μL) and acetic anhydride (10 μL) were added and reacted overnight. The reaction solution was placed in a dialysis membrane with a molecular weight cutoff of 3500 and dialyzed for 2 days, after which the product was recovered by lyophilization.

[0096] In this way, we obtained hyperbranched polymers (Cy5-8arm-PyBA, Cy5-8arm-FPBA, Cy5-8arm-PBA, Cy5-8arm-APBA, Cy5-8arm-Phenyl, Cy5-8arm-Ace) in which Cy5 was bound to one of the residues shown below the eight terminal amino groups of the 8-arm-PEG shown below. Note that the -NH- of the amide bond at the bond site of each residue originates from the terminal amino group of the 8-arm-PEG. [ka]

[0097] Regarding the hyperbranched polymer obtained in 2-2 to 2-7 above, NMR analysis was performed under the following conditions. As a result, the average number of Cy5 bindings per polymer molecule was 0.9, and the average number of each ligand was 6.5 or more. Also, regarding the hyperbranched polymer obtained in 2-2 to 2-6 above, FCS analysis was performed under the following conditions. As a result, the particle diameter of each hyperbranched polymer was within the range of 8 nm to 10 nm. <NMR Analysis> The hyperbranched polymer was dissolved in D2O at a concentration of 10 mg / mL to obtain a measurement sample. The above measurement sample was subjected to NMR measurement using an NMR apparatus (ECS-400) manufactured by JEOL Ltd. to calculate the number of introduced ligands. The measurement conditions were set at 25 °C and 64 integration times. <FCS Analysis> The hydrodynamic diameter of the hyperbranched polymer was measured using an FCS apparatus (MF20) manufactured by Olympus Corporation. The hyperbranched polymer was dissolved in D-PBS(-) at a concentration of 5 mg / L. As a standard dye, Cy5 dye was dissolved in D-PBS(-) at a concentration of 10 nM, and the diffusion times of the hyperbranched polymer and Cy5 dye were measured. The hydrodynamic diameter was calculated based on the obtained diffusion times.

[0098] 3. In Vivo Kinetics Evaluation The above hyperbranched polymer was dissolved in D-PBS(-) to a concentration of 10 μM, and 200 μL was administered to a mouse (C57 / BL6, female, 7 weeks old) via the tail vein. After 24 hours of administration, the mouse was dissected, and the accumulation rate of the hyperbranched polymer in each organ was calculated from the fluorescence intensity of Cy5 in the blood and the crushed solution of the sampled organs (tissues). The results are shown in Figure 4 (in the figure, the bars indicate the average values for n = 3, and the error bars indicate the standard deviation).

[0099] As shown in Figure 4, Cy5-8arm-PyBA, Cy5-8arm-FPBA, Cy5-8arm-PBA, and Cy5-8arm-APBA, which had boronic acid groups introduced, all showed higher cardiac accumulation than Cy5-8arm-Ace, which had no ligand. The enhanced accumulation effect of Cy5-8arm-PyBA, Cy5-8arm-FPBA, and Cy5-8arm-PBA was particularly notable. On the other hand, Cy5-8arm-Phenyl, which had a phenyl group introduced, did not show any enhanced cardiac accumulation. Furthermore, Cy5-8arm-PyBA, Cy5-8arm-FPBA, and Cy5-8arm-PBA also showed a significant effect of increasing accumulation in the diaphragm.

[0100] [Experimental Example 2: Drug delivery particles containing graft polymers as DDS carrier constituent molecules] 1. Raw materials Azido-PEG3-amine (purchased from Tokyo Chemical Industry Co., Ltd., used as is) N,N-dimethylformamide (DMF) (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used after distillation with calcium hydride or as is) Dimethyl sulfoxide (DMSO) (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) Dichloromethane (purchased from Fujifilm Wako Pure Chemical Industries, used after distillation with calcium hydride) β-Benzyl-L-aspartic acid N-carboxylic anhydride (NCA-BLA) (purchased from Chuo Kaseihin, used as is) Product name: "SUNBRIGHT MEPA-20H" (Methoxy-PEG-amine with an average molecular weight of 2,000, purchased from NOF Corporation, used as is) Product name: "SUNBRIGHT BO-020EA" (Boc-protected amine-PEG-amine with an average molecular weight of 2,000, purchased from NOF Corporation, used as is) 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) N-hydroxysuccinimide (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) Diethyl ether (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) Acetonitrile (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) Sodium hydroxide (purchased from Fujifilm Wako Pure Chemical Industries, used as is) Sodium bicarbonate (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) Trifluoroacetic acid (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is) Sulfo-Cy5-DBCO (purchased from Lumiprobe, used as is) D-PBS(-) (purchased from Fujifilm Wako Pure Chemical Industries, Ltd., used as is)

[0101] 2-1. Synthesis of PBLA and polyaspartic acid (PAsp) 1,000 mg of NCA-BLA was weighed into a round-bottom flask and dissolved in 2 mL of DMF. In a separate round-bottom flask, 724 μL of azide-PEG3-amine solution diluted 100-fold with dichloromethane was added to 18 mL of dichloromethane and mixed. The entire NCA-BLA solution was added to the azide-PEG3-amine solution and stirred for 3 days at 35°C. All reactions were carried out under an argon atmosphere, and all organic solvents used were distilled prior to use. After the reaction, the reaction solution was added dropwise to 400 mL of diethyl ether to precipitate PBLA. The precipitate was then collected by suction filtration and dried under reduced pressure. The degree of polymerization of the resulting PBLA was measured by NMR, and calculated to be 101. 100 mg of PBLA was suspended in 1 mL of acetonitrile, 4.9 mL of 0.5 M sodium hydroxide solution was added, and the mixture was stirred overnight at room temperature. The reaction mixture was then placed in a dialysis membrane (molecular weight cutoff: 6,000-8,000) and dialyzed twice with pure water, twice with 0.01 M hydrochloric acid, and twice with pure water (for at least 2 hours each). Polyaspartic acid (PAsp) was obtained by lyophilization. All dialysis was performed in a refrigerator.

[0102] 2-2.Synthesis of PyBA-PEG-amine DMF (5 mL) was added to BO-020EA (250 mg), PyBA (104 mg), and sodium bicarbonate (20 mg). After thorough stirring, DMT-MM (180 mg) was added and the mixture was allowed to react overnight. The insoluble material was removed from the reaction mixture using a 0.45 μm filter, and the mixture was added dropwise to 100 mL of diethyl ether and dried to obtain a white powder. Dichloromethane (2.5 mL) and trifluoroacetic acid (250 μL) were added, and the mixture was allowed to react overnight at room temperature. The insoluble material was removed using a 0.45 μm filter, and the mixture was added dropwise to 50 mL of diethyl ether and dried to obtain a white powder (PyBA-PEG-amine).

[0103] 2-3.Synthesis of FPBA-PEG-amine Methanol (5 mL) was added to BO-020EA (250 mg) and FPBA (115 mg), and after thorough stirring, DMT-MM (180 mg) was added and the mixture was allowed to react overnight. The insoluble material was removed from the reaction mixture using a 0.45 μm filter. After drying under reduced pressure, the mixture was redissolved in dichloromethane (5 mL). After removing the insoluble material using a 0.45 μm filter, the mixture was added dropwise to 100 mL of diethyl ether and dried to obtain a white powder. Dichloromethane (2.5 mL) and trifluoroacetic acid (250 μL) were added, and the mixture was allowed to react overnight at room temperature. The insoluble material was removed using a 0.45 μm filter. The mixture was added dropwise to 50 mL of diethyl ether and dried to obtain a white powder (FPBA-PEG-amine).

[0104] 2-4. Introduction of various PEGs (MEPA-20H, PyBA-PEG-amine, FPBA-PEG-amine) into PAsp Introduction of MEPA-20H: PAsp (3 mg) was weighed and dissolved in 1.5 mL of a 50% mixture of DMF and DMSO. In a separate sample bottle, 150 mg of MEPA-20H was weighed and dissolved in 3 mL of DMF. N-hydroxysuccinimide (14.65 mg) and EDC (4 mg) were added to the PAsp solution and stirred at room temperature for 30 minutes. 2.36 mL of PEG solution was added and allowed to react overnight. PyBA-PEG-amine and FPBA-PEG-amine were also introduced in a similar manner.

[0105] The reaction solution was placed in a dialysis membrane with a molecular weight cut-off of 12,000 - 14,000 and dialyzed twice with PBS. Then, it was purified by ultrafiltration (molecular weight cut-off 30,000) and recovered by lyophilization.

[0106] The number of PEG introduced into PAsp was calculated by measuring the decrease in the initial peak area of each PEG using GPC. From the GPC results, the average number of introduced PEG was calculated to be 52 for MEPA-20H, 50 for PyBA-PEG-amine, and 57 for FPBA-PEG-amine.

[0107] 2 - 5. Introduction of fluorescent label (Cy5) into each graft polymer The graft polymer (15 mg) with various PEG introduced was dissolved in pure water, and 7 μL of a Sulfo-Cy5-DBCO solution adjusted to a concentration of 25 mg / mL with DMSO was added. After stirring, it was left standing at -20 °C overnight and then redissolved in the refrigerator. The resulting solution was placed in a dialysis membrane with a molecular weight cut-off of 12,000 - 14,000 and dialyzed 4 times with pure water, and then recovered by lyophilization.

[0108] In the above manner, three types of graft polymers (polyPyBA, polyFPBA, and polyPEG) shown below were obtained (L represents a divalent linker, m is 101, the number of introduced R1 is 50 for PyBA-PEG, 57 for FPBA-PEG, and 52 for PEG, and each amino acid residue exists randomly).

Chemical formula

[0109] Regarding each graft polymer obtained in 2 - 5 above, DLS analysis was performed under the following conditions. As a result, the particle size of each graft polymer was within the range of 12 nm to 20 nm. <DLS analysis> The hydrodynamic diameter of the grafted polymer was measured using a DLS device (Mobius) manufactured by Wyatt Technology Co., Ltd. The grafted polymer was dissolved in D-PBS(-) at a concentration of 5 mg / mL, and the hydrodynamic diameter was measured.

[0110] 3. Pharmacokinetic evaluation The above graft polymer was dissolved in D-PBS(-) to a concentration of 10 μM, and 200 μL of the solution was administered via the tail vein to mice (C57 / BL6, female, 7 weeks old). 48 hours after administration, the mice were dissected, and the accumulation rate of the graft polymer in each organ was calculated from the Cy5 fluorescence intensity in the blood and in the homogenate of the collected organs (tissues). The results are shown in Figures 5A and 5B (in the figures, the bars indicate the average value (n = 3), and the error bars indicate the standard deviation). Note that Figure 5B shows the relative accumulation amount when the accumulation amount of polyPEG is set to 1.

[0111] As shown in Figures 5A and 5B, both polyPyBA and polyFPBA grafted with PEG containing boronic acid groups showed higher accumulation in the heart and diaphragm than polyPEG grafted with PEG not containing boronic acid groups. [Industrial Applicability]

[0112] The drug delivery particles of the present invention can be suitably used in the delivery of drugs targeted to the heart and / or diaphragm in the medical field.

Claims

1. Particles for drug delivery to the heart and / or diaphragm having boronic acid groups as targeting moieties.

2. The drug delivery particle of claim 1 , wherein the boronic acid group comprises a phenylboronic acid group or a pyridineboronic acid group.

3. The drug delivery particle according to claim 1, wherein the particle diameter is 100 nm or less.

4. The drug delivery particle according to claim 1 , comprising a drug transporter component molecule to which the boronic acid group and a drug are bound.

5. The drug delivery particle according to claim 4 , wherein the drug transporter component molecule is a linear polymer, a branched polymer, or a graft polymer.

6. each of which contains a complex of a specific number of drug transporter component molecules and a drug; The drug delivery particle according to claim 1 , wherein the boronic acid group is bound to at least one of the drug transporter component molecule and the drug.

7. The drug delivery particle according to claim 1 , comprising a micelle or vesicle containing a drug transporter component molecule to which the boronic acid group is bound.

8. further comprising a drug, The drug delivery particle according to claim 7 , wherein the drug is encapsulated in the micelle or the vesicle.

9. The drug delivery particle of claim 1 , comprising a drug having the boronic acid group attached thereto.

10. The drug delivery particle according to claim 1, which is used to treat a disease requiring delivery of a drug to the heart and / or diaphragm.

11. A pharmaceutical composition comprising the drug delivery particles of claim 1.

12. The use of boronic acid groups to target drugs or drug transporters to the heart and / or diaphragm.

13. Use of a boronic acid compound for producing the drug delivery particle of claim 1.

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