WO2017164334A1 - Boron cluster bonded peptide compound - Google Patents

Boron cluster bonded peptide compound Download PDF

Info

Publication number
WO2017164334A1
WO2017164334A1 PCT/JP2017/011843 JP2017011843W WO2017164334A1 WO 2017164334 A1 WO2017164334 A1 WO 2017164334A1 JP 2017011843 W JP2017011843 W JP 2017011843W WO 2017164334 A1 WO2017164334 A1 WO 2017164334A1
Authority
WO
WIPO (PCT)
Prior art keywords
group
amino acid
compound
salt
peptide compound
Prior art date
Application number
PCT/JP2017/011843
Other languages
French (fr)
Japanese (ja)
Inventor
片岡　一則
鵬 米
オラシオ カブラル
柳衛　宏宣
デウィ ノフリアナ
西山　伸宏
中村　浩之
宏泰 武元
貴大 野本
渉 黒澤
吏紗 姥貝
吉朗 北原
泰世 須賀
Original Assignee
公益財団法人 川崎市産業振興財団
国立大学法人 東京大学
国立大学法人 東京工業大学
味の素株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 公益財団法人 川崎市産業振興財団, 国立大学法人 東京大学, 国立大学法人 東京工業大学, 味の素株式会社 filed Critical 公益財団法人 川崎市産業振興財団
Priority to JP2018507416A priority Critical patent/JPWO2017164334A1/en
Publication of WO2017164334A1 publication Critical patent/WO2017164334A1/en

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/22—Boron compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic System
    • C07F5/02—Boron compounds
    • C07F5/05—Cyclic compounds having at least one ring containing boron but no carbon in the ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G81/00—Macromolecular compounds obtained by interreacting polymers in the absence of monomers, e.g. block polymers

Definitions

  • It relates to novel boron-containing compounds useful for boron neutron capture therapy, and in particular, to peptide compounds into which boron clusters are introduced.
  • NCT Neutron capture therapy
  • boron neutron capture therapy has attracted attention as a minimally invasive treatment method that exhibits an excellent therapeutic effect on solid cancers that are difficult to perform surgery and has less burden on patients.
  • NCT irradiates the affected area with low-energy thermal neutrons or epithermal neutrons after administering molecules containing elements with large reaction cross sections with neutrons such as 10 B and 157 Gd into the body.
  • cytotoxic radiation ⁇ rays or the like
  • BSH mercaptoundecahydrodedecaborate
  • BSH is rapidly excreted and has poor blood retention and tumor retention.
  • a thermal neutron beam or an epithermal neutron beam while administering a very high concentration compound into the body.
  • cancer cells may not be able to be completely cured due to insufficient concentration of 10 B in the tumor, damage to normal tissue may occur, thermal neutron radiation or There is a problem that applicable diseases are limited because pinpoint irradiation with neutron beams is necessary.
  • the retention in blood is increased, and enhanced permeability and retention (EPR) effect (increased permeability of tumor blood vessels and undeveloped lymphatic system in cancer tissues)
  • EPR enhanced permeability and retention
  • a cationic dendrimer having PEG has been developed [B. Qualmann et al. , Angew. Chem. Int. Ed. , 35, 909-911 (1996)], and is highly membrane permeable due to the cationic nature of many amino groups, nonspecifically adsorbing to biomolecules, and difficult to selectively deliver to cancer. In addition, it cannot be expected to be uniformly distributed in the tumor. Furthermore, it may be recognized that the strong positive charge is recognized as a foreign substance in the living body. In addition, when used in BNCT, unlike a normal medicine, a large dose is required. However, a polymer of a basic amino acid is known for cytotoxicity and is not suitable for a large dose.
  • An object of the present invention is to provide a novel boron-containing compound that exhibits excellent cancer accumulation due to the EPR effect, and high penetrability in tumor tissue by not forming nanoparticles, and is efficiently taken into cancer cells. Thus, it is to realize effective treatment of tumor diseases in boron neutron capture therapy.
  • the present inventors have obtained a peptide compound having a boron cluster such as polyether and BSH, and an amino acid selected from a neutral amino acid and an acidic amino acid as a constituent amino acid.
  • a peptide compound having a boron cluster such as polyether and BSH
  • an amino acid selected from a neutral amino acid and an acidic amino acid as a constituent amino acid.
  • each R 1 independently represents an ⁇ selected from a neutral amino acid and an acidic amino acid substituted with a monovalent group consisting of a boron cluster or a monovalent group to which it is bonded via a linker structure.
  • R 1 s are each independently represented by formula (B1):
  • Each R 1 independently represents the formula (R1a):
  • each R a is independently a hydrogen atom, a carboxy group optionally substituted with an alkyl group having 1 to 6 carbon atoms, or an alkyl group having 1 to 6 carbon atoms. good hydroxy group, an alkyl mercapto group optionally substituted with a group with a carbon number of 1-6, an aryl group, or an alkyl group having 1 to 6 carbon atoms having a carbon number of 6 ⁇ 14;
  • Q 2 is Independently, —O—, —S—, —NH—, —SO 2 —, —CO—, —NHCO—, —CONH—, —OCO—, —COO—, —SCO—, —COS—, —NHCONH—, —NHCSNH—, —OCONH—, —NHCOO—, —SS—, —Ph—, —OPhO—, —OPh—, —PhO—, —SPhS—, —SPhS
  • the ⁇ -amino acid of R 1 is glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, asparagine, glutamine, phenylalanine, tyrosine, aspartic acid, glutamic acid, homoserine, norleucine, norvaline, thyronine,
  • the peptide compound or salt thereof according to any one of [4] to [6] above, selected from the group consisting of citrulline, ⁇ -aminobutyric acid, homocysteine and penicillamine.
  • linker structure is a divalent group consisting of 1 to 200 main chain constituent atoms selected from a carbon atom, an oxygen atom, a sulfur atom and a nitrogen atom Or a salt thereof.
  • the linker structure has the formula (L ′):
  • each R a is independently a hydrogen atom, a carboxy group optionally substituted with an alkyl group having 1 to 6 carbon atoms, or an alkyl group having 1 to 6 carbon atoms.
  • P 2 and Q 2 are each independently —O—, —S—, —NH—, —SO 2 —, —CO—, —NHCO—, —CONH—, —OCO—, —COO—, —SCO—, — COS-, -NHCONH-, -NHCSNH-, -OCONH-, -NHCOO-, -SS-, -Ph-, -OPhO-, -OPh-, -PhO-, -SPhS-, -SP
  • Z represents —O—, —NH— or —S— when the substitution target is a carboxy group, and —CO—, —CONH— or —COO— when the substitution target is an amino group
  • R d And R e each independently represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms
  • s is each independently an integer of 1 to 10
  • t is independently 2 to 10
  • Each is an integer
  • u is each independently an integer of 1 or more
  • *** represents a binding site.
  • the boron cluster is selected from the group consisting of a borohydride compound, a compound in which one or more boron atoms are each independently substituted with a carbon atom or a nitrogen atom, and a metal complex thereof.
  • a borohydride compound a compound in which one or more boron atoms are each independently substituted with a carbon atom or a nitrogen atom, and a metal complex thereof.
  • R 1 is each independently an ⁇ -amino acid selected from a neutral amino acid and an acidic amino acid substituted with a monovalent group consisting of a boron cluster or a monovalent group to which it is bonded via a linker structure
  • Showing the chain R 2 each independently represents a side chain of an ⁇ -amino acid selected from a neutral amino acid and an acidic amino acid, which may have a substituent,
  • the side chain of the ⁇ -amino acid of R 1 and R 2 may be combined with the carbon atom to which it is attached and the nitrogen atom adjacent to the carbon atom to form a ring.
  • X 1 and Y 2 when c is 1 are each independently (1) a hydrogen atom, (2) an alkyl group having 1 to 6 carbon atoms, (3) an alkylcarbonyl group having 2 to 7 carbon atoms, or (4) a monovalent group comprising a polyether, X 2 and Y 1 when c is 1 are each independently (1) a hydroxy group which may be substituted with an alkyl group having 1 to 6 carbon atoms, (2) an amino group which may be substituted with an alkyl group having 1 to 6 carbon atoms, or (3) a monovalent group comprising a polyether, Y 1 and Y 2 in the case where c is 2 or more each represent a c-valent group composed of a polyether; In formula (Ia), at least one of the substituents on
  • Y 1 and Y 2 are represented by the formula (P1):
  • R d and R e is independently a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, s is independently an integer of 1 to 10, and t is independently an integer of 2 to 10; Each u is independently an integer of 1 or more, and *** represents a binding site.
  • X 1 is a hydrogen atom
  • X 2 is a hydroxy group which may be substituted with an alkyl group having 1 to 6 carbon atoms, or an alkyl group having 1 to 6 carbon atoms.
  • a pharmaceutical composition comprising the peptide compound or a salt thereof according to any one of [1] to [32] above, and a pharmaceutically acceptable carrier.
  • a method for treating a tumor disease by boron neutron capture therapy comprising administering a necessary amount of the peptide compound or salt thereof according to any one of [1] to [32] to a mammal.
  • the compound of the present invention exhibits excellent cancer accumulation due to the EPR effect, and high penetrability in tumor tissues due to non-nanoparticulation, and is efficiently incorporated into cancer cells.
  • Neutron capture therapy can treat tumor diseases more effectively.
  • concentration of the compound of Examples 1-3 and the comparative examples 1 and 2 and 10 B-BSH in the cytotoxicity test with respect to HUVEC is shown.
  • 2 is a graph showing the cell viability according to the concentrations of the compounds of Examples 1 to 3 and Comparative Examples 1 and 2 and 10 B-BSH in the cytotoxicity test for C26 cancer cells.
  • capture amount in the C26 cancer cell of the compound of Examples 1-3 and the comparative examples 1 and 2, and 10 B-BSH is shown.
  • the vertical axis of the graph shows the relative cellular uptake of each compound when the amount of cellular uptake for 1 hour after addition of 10 B-BSH to C26 cancer cells is 1.
  • the image by the confocal laser scanning microscope (CLSM) of the C26 cancer cell which added the compound which fluorescently labeled the compound of Example 1 with Alexa488 (trademark) is shown.
  • capture of the compound whose counter cation of the compound of Example 7 and the compound of the synthesis example 21 is a 2 cesium cation is shown.
  • 2 is a graph relating to plasma concentration transitions after administration of the compounds of Examples 1 to 3 or Comparative Example 1 or 2 or 10 B-BSH to mice transplanted subcutaneously with C26 cancer cells.
  • the vertical axis of the graph shows what percentage of the dose (ID) delivered per ml of plasma.
  • FIG. 3 shows a graph regarding the change in the amount of accumulation in tumor cells after administering the compound of Examples 1 to 3 or Comparative Example 1 or 2 or 10 B-BSH to mice transplanted subcutaneously with C26 cancer cells.
  • the vertical axis of the graph shows what percentage of the dose (ID) delivered per gram of tumor.
  • 2 is a graph relating to the change in the amount of accumulation in tumor cells after the compound of Example 7 or natural B-BSH was administered to mice subcutaneously transplanted with CT26 cancer cells.
  • the vertical axis of the graph shows what percentage of the dose (ID: injected dose) was delivered per 1 g tumor.
  • the graph regarding the relative tumor volume transition when the compound of Example 1 or 10 B-BSH is administered to a mouse transplanted with C26 cancer cells subcutaneously and after irradiation with thermal neutrons for 1 hour or without irradiation is shown.
  • mouth transplanted subcutaneously with the C26 cancer cell, and a thermal neutron ray is irradiated for 1 hour, or is not irradiated is shown.
  • mouth about the compound of Example 1 and a liposome formulation is shown.
  • the upper left figure shows hematoxylin-eosin staining of C26 tumor tissue.
  • the lower left figure shows the C26 tumor tissue distribution of the compound of Example 1 fluorescently labeled with Alexa647 (registered trademark) and Doxil (registered trademark).
  • the middle panel shows hematoxylin-eosin staining of BxPC3 tumor tissue.
  • the right figure shows the BxPC3 tumor tissue distribution of the compound of Example 1 fluorescently labeled with Alexa647 (registered trademark) and Doxil (registered trademark).
  • a compound obtained by fluorescently labeling the compound of Example 1 with Alexa 647 (registered trademark) is shown in green, Doxil (registered trademark) in red, and a portion where both compounds coexist is shown in yellow.
  • the present invention provides a peptide compound having a boron cluster and a polyether and having an amino acid selected from a neutral amino acid and an acidic amino acid as a constituent amino acid (hereinafter referred to as “the peptide compound of the present invention”) or a salt thereof.
  • peptide means a peptide in which two or more amino acids are bound by peptide bonds.
  • the number of amino acid residues of the peptide is preferably 3 or more, more preferably 5 or more, and still more preferably 10 or more.
  • the number of amino acid residues of the peptide is preferably 200 or less, more preferably 150 or less, and still more preferably 100 or less.
  • the “peptide compound” means a peptide which may have an arbitrary substituent.
  • the peptide of the peptide compound is preferably a linear peptide, not a peptide dendrimer or a branched peptide, from the viewpoint of binding many boron clusters.
  • amino acid means a known amino acid of natural type or non-natural type.
  • neutral amino acid means a known neutral amino acid of natural type or non-natural type.
  • acidic amino acid means a known acidic amino acid of natural type or non-natural type. For example, aspartic acid and glutamic acid.
  • the constituent amino acids of the peptide preferably have an active group such as carboxy group, hydroxy group, mercapto group in the side chain of serine, threonine, cysteine, tyrosine, aspartic acid, glutamic acid, homoserine, homocysteine, penicillamine, hydroxyproline, etc.
  • an active group such as carboxy group, hydroxy group, mercapto group in the side chain of serine, threonine, cysteine, tyrosine, aspartic acid, glutamic acid, homoserine, homocysteine, penicillamine, hydroxyproline, etc.
  • amino acids may be D-type, L-type or a mixture thereof.
  • the peptide is a polyamino acid. That is, all the constituent amino acids of the peptide are the same amino acid.
  • the peptide compound has at least one amino acid residue having a boron cluster bonded to a side chain.
  • the ratio of the number of amino acid residues in which the boron cluster is bonded to the side chain to the number of amino acid residues of the peptide is preferably 0.1 or more, more preferably 0.2 or more, and still more preferably 0. .3 or more.
  • amino acid residue is a unit of a constituent amino acid of a peptide, and means a group excluding OH at the C-terminal of the amino acid and / or H at the N-terminal.
  • side chain of an amino acid means a branched chain that does not constitute the main chain when constituting a peptide.
  • the “boron cluster” means an ionic or nonionic known substance having a structure obtained mainly by combining a plurality of boron atoms and combining them into 3 to 20 boron atoms.
  • a boron cluster having atoms is preferable, a boron cluster having 8 to 20 boron atoms is more preferable, and a boron cluster having 10 to 12 boron atoms is particularly preferable.
  • the boron cluster may contain a carbon atom, a nitrogen atom, an oxygen atom, a sulfur atom, and the like as a skeleton atom constituting the boron cluster.
  • the number of other atoms such as carbon atom, nitrogen atom, oxygen atom and sulfur atom is preferably 0 to 5, and more preferably 0 to 2.
  • the boron cluster may be a metal complex with a transition metal such as nickel or cobalt.
  • the boron cluster is preferably ionic and preferably water-soluble.
  • Examples of the boron cluster include [B 3 H 8 ] ⁇ M + , [B 6 H 6 ] 2 ⁇ M 2+ , [B 7 H 7 ] 2 ⁇ M 2+ , [B 8 H 8 ] 2 ⁇ M 2+ , [B 9 H 9 ] 2 ⁇ M 2+ , [B 10 H 10 ] 2 ⁇ M 2+ , [B 10 H 13 ] ⁇ M + , [B 11 H 11 ] 2 ⁇ M 2+ , [B 11 H 14 ] ⁇ Boron hydride compounds such as M + , [B 12 H 12 ] 2 ⁇ M 2+ , [B 20 H 18 ] 4 ⁇ M 4+ , wherein M + , M 2+ and M 4+ each represent a cation.
  • boron clusters represented by the following formula.
  • ⁇ represents BH, and the other symbols are as defined above.
  • boron cluster represented by the following formula is preferable.
  • M + , M 2+ or M 4+ is not particularly limited as long as it is a pharmaceutically acceptable cation.
  • M + , M 2+ and M 4+ may be a single component or a combination of two or more components, and each cation may be monovalent or polyvalent. .
  • alkali metal ions for example, sodium ions, potassium ions, cesium ions, etc.
  • alkaline earth metal ions for example, magnesium ions, calcium ions, etc.
  • amine ions for example, tetramethylammonium ions
  • a cation selected from monovalent or polyvalent ammonium ions of polyamines for example, 1,4-diaminobutane, spermine, spermidine
  • alkali metal ions for example, sodium ions, potassium ions, cesium ions, etc.
  • alkaline earth metal ions for example, magnesium ions, calcium ions, etc.
  • amine ions for example, tetramethylammonium ions
  • a cation selected from monovalent or polyvalent ammonium ions of polyamines for example, 1,4-diaminobutane, spermine, spermidine
  • alkali metal ions for example, 1,4-diaminobutane, spermine, spermidine
  • the 10 B isotope ratio of the boron cluster is preferably 19% or more, more preferably 50% or more, and still more preferably 80% or more.
  • the boron cluster is bonded to, for example, a constituent amino acid of the peptide directly or via a linker structure.
  • the boron cluster is preferably bonded to one or more side chains of the constituent amino acids of the peptide, more preferably 1 to 3, and particularly preferably 1.
  • the boron cluster is a monovalent group (meaning a group formed by removing any one H of a boron cluster from the side chain of an amino acid, and hereinafter referred to as “monovalent group consisting of a boron cluster”). More preferably, it is covalently bonded directly or via a linker structure.
  • the boron cluster (or linker structure) is attached, for example, by substitution with any H or OH on the carbon, nitrogen or sulfur atom of the side chain of the amino acid, and the active group of the amino acid. More preferably, it is bonded to a group selected from H and OH (for example, OH of a carboxy group and H of a hydroxy group and a mercapto group), and is bonded to a OH of a carboxy group. Is more preferable.
  • Examples of the bond that the boron cluster (or linker structure) can form with the carboxy group include an amide bond (—CONH—), an ester bond (—COO—), a thioester bond (—COS—), and the like. Preferably, it is an amide bond.
  • Examples of the bond that can be formed with the hydroxy group include an ester bond (—OCO—) and a urethane bond (—OCONH—).
  • Examples of the bond that can be formed with the mercapto group include a thioester bond (—SCO—, —COS—, disulfide bond (—SS—) and the like.
  • the ratio of the number of bonds of the boron cluster to the number of amino acid residues of the peptide is preferably 0.1 or more, more preferably 0.2 or more, and further preferably 0.3 or more.
  • the monovalent group consisting of boron clusters is preferably formula (B1) to formula (B15):
  • linker structure in the present specification is not particularly limited as long as it is a pharmaceutically acceptable structure capable of linking a boron cluster and an amino acid side chain.
  • examples thereof include a divalent group consisting of main chain constituent atoms selected from carbon atoms, oxygen atoms, sulfur atoms and nitrogen atoms (preferably 1 to 200, more preferably 1 to 30).
  • the “linker structure” is preferably a divalent group represented by the following formula (L):
  • P 1 and Q 1 are each independently —O—, —S—, —NH—, —SO 2 —, —CO—, —NHCO—, —CONH—, —OCO—, —COO—, —SCO—, —COS—, —NHCONH—, —NHCSNH—, —OCONH—, — NHCOO -, - SS -, - NHCOS -, - SCONH -, - NHCH 2 CONH -, - NHCOCH 2 NH -, - NHCH 2 COO -, - OCOCH 2 NH -, - OCH 2 CONH -, - NHCOCH 2 O- , -OCH 2 COO -, - OCOCH 2 O -, - SCH 2 CONH -, - NH, NHCOCH 2 O-, -OCH 2 COO -, - OCOCH 2 O -, -
  • Such a linker structure can be introduced by selecting various known reactions and known linker structures in order to link boron clusters and amino acid side chains.
  • Such linker structures are well known and are described, for example, in S.I. Manabeand M.M. Yokoyama, Drug Delivery System, 30-3: 247-250 (2015), N.I. Jain et al. Pharm Res, 32: 3256-3540 (2015), J. Am. R. McCombs et al. The AAPS Journal, 17 (2): 339-351 (March 2015), J.A. Khandare et al. , Prog. Polym. Sci. 31: 359-397 (2006).
  • the “linker structure” is preferably a divalent group represented by the following formula (L ′):
  • Each R a is independently a hydrogen atom, a carboxy group optionally substituted with an alkyl group having 1 to 6 carbon atoms, a hydroxy group optionally substituted with an alkyl group having 1 to 6 carbon atoms, A mercapto group optionally substituted with an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 14 carbon atoms, or an alkyl group having 1 to 6 carbon atoms;
  • P 2 and Q 2 are each independently —O—, —S—, —NH—, —SO 2 —, —CO—, —NHCO—, —CONH—, —OCO—, —COO—, — SCO—, —COS—, —NHCONH—, —NHCSNH—, —OCONH—, —NHCOO—, —SS—, —Ph—, —OPhO—, —OPh—, —PhO—, —SPhS—,
  • Such a linker structure can be introduced by selecting various known reactions and known linker structures in order to link boron clusters and amino acid side chains.
  • heterocycle refers to a ring containing 1 to 4 heteroatoms selected from the group consisting of an oxygen atom, a sulfur atom and a nitrogen atom in addition to a carbon atom as a ring constituent atom.
  • specific examples include hetetidine, pyrrolidine, piperidine, morpholine, thiomorpholine, piperazine, oxazolidine, thiazolidine, dihydrothiopyran, imidazolidine, oxazoline, thiazoline, imidazoline.
  • 3-8 membered non-aromatic heterocycles furan, thiophene, pyridine, pyrimidine, pyridazine, pyrazine, pyrrole, imidazole, pyrazole, thiazole, isothiazole, oxazole, iso Examples thereof include 5- or 6-membered aromatic heterocycles such as oxazole, oxadiazole, thiadiazole, triazole, tetrazole, and triazine.
  • an alkyl group having 1 to 6 carbon atoms means a linear, branched or cyclic saturated hydrocarbon group having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, Examples include isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, cyclopentyl, cyclohexyl and the like.
  • aryl group having 6 to 14 carbon atoms means a cyclic aromatic hydrocarbon group having 6 to 14 carbon atoms, and examples thereof include phenyl, 1-naphthyl, 2-naphthyl and the like. It is done.
  • Each R a is preferably a hydrogen atom.
  • P 1 and P 2 are each preferably —CO—, —CONH— or —COO— when the binding target in the side chain of the amino acid is O or S (for example, a hydroxy group or a mercapto group),
  • the binding target in the side chain of an amino acid is CO (for example, a carboxy group)
  • it is preferably —O—, —NH— or —S—.
  • Q 2 is preferably each independently —NHCO—, —CONH—, —O—, —S—, —SS— or —heterocycle—.
  • the “polyether” means a linear or branched polymer having an ether bond in the main chain, preferably a polyalkylene glycol unit (preferably a polyethylene glycol (PEG) unit). Is a linear or branched water-soluble polymer.
  • the polyalkylene glycol unit is preferably contained in the polyether in an amount of 60% by weight or more, more preferably 80% by weight or more, and further preferably 90% by weight or more.
  • the atoms of the terminal part or the chain part of the main chain of the polyether may be atoms other than oxygen atoms and carbon atoms (for example, sulfur atoms, nitrogen atoms, etc.).
  • the number average molecular weight (Mn) of “polyether” in the present specification is preferably less than 50,000, and more preferably less than 30,000. Further, the number average molecular weight (Mn) of the “polyether” is preferably 1,000 or more, more preferably 10,000 or more.
  • the molecular weight distribution (weight average molecular weight (Mw) / number average molecular weight (Mn)) of the “polyether” is preferably 3 or less, more preferably 2 or less.
  • the polyether is an active group of a constituent amino acid of the peptide (for example, a carboxy group, a hydroxy group and a mercapto group on the side chain of the constituent amino acid of the peptide compound, and an amino group and C at the N terminal of the peptide compound. It is preferably bonded to one or more groups selected from (terminal carboxy group), and particularly preferably bonded to either the N-terminal amino group or the C-terminal carboxy group of the peptide compound.
  • a constituent amino acid of the peptide for example, a carboxy group, a hydroxy group and a mercapto group on the side chain of the constituent amino acid of the peptide compound, and an amino group and C at the N terminal of the peptide compound.
  • the polyether is bonded to the peptide as a monovalent group (meaning a group formed by removing any one H or OH of the polyether, and hereinafter referred to as “monovalent group made of polyether”). Or a polyvalent group (meaning a group formed by removing two or more groups selected from any H and OH of a polyether, and hereinafter referred to as a “c-valent group comprising a polyether” ( c is an integer of 2 or more))) and may be bonded to a plurality of peptides.
  • the monovalent group made of polyether and the c-valent group made of polyether are each preferably one of OH of the carboxy group and H of the amino group, hydroxy group and mercapto group at the end of the polyether chain. Or it is group which removes c pieces.
  • the bond that the polyether can form with an amino group, a carboxy group, a hydroxy group, and a mercapto group, the same bonds as those listed above as the bonds that the boron cluster (or linker structure) can form respectively. Is mentioned.
  • Z represents —O—, —NH—, —S—, —CO—, —CONH—, —COO—, —NHCO—, —CONH—, —OCO—, —COO—, —SCO—, —COS—.
  • -NHCONH-, -NHCSNH-, -OCONH-, -NHCOO- or -SS- R d and R e each independently represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, s each independently represents an integer of 1 to 10 (preferably 1 to 4), t is each independently an integer of 2 to 10 (preferably 2 to 4); u is each independently an integer of 1 or more, *** indicates a binding site.
  • the group represented by these is mentioned. Among them, a group represented by the above formula (P1) is preferable.
  • R d is preferably a hydrogen atom.
  • R e is preferably an alkyl group having 1 to 6 carbon atoms, more preferably methyl.
  • the total of u is preferably an integer of 1,000 or less, more preferably an integer of 600 or less.
  • Z is preferably —CO—, —CONH— or —COO— when the binding target is O, N, NH or S (for example, a hydroxy group, an amino group or a mercapto group), and the binding target is CO (for example, In the case of a carboxy group, it is preferably —O—, —NH— or —S—.
  • At least one of the active groups of the constituent amino acids of the peptide is substituted with a monovalent group consisting of a polyether. More preferably, the C-terminal carboxy group of the peptide compound and / or the N-terminal amino group of the peptide compound are substituted with a monovalent group comprising a polyether.
  • the carboxy group at the C-terminal of the peptide compound and / or the amino group at the N-terminal of the peptide compound are represented by the formula (P1) (wherein Z is —O— when the substitution target is a carboxy group, When —NH— or —S— is substituted with an amino group, it represents —CO—, —CONH— or —COO—, and the other symbols are as defined above. ing.
  • an amino acid residue having a boron cluster (or linker structure) bonded to a side chain is represented by the following formula (1).
  • ⁇ -amino acid means an amino acid in which one carboxyl group and one amino group are bonded to one carbon atom (referred to as ⁇ -carbon).
  • the ⁇ -amino acid may be D-form, L-form or a mixture thereof.
  • the “side chain of ⁇ -amino acid” refers to a group corresponding to R 3 when the ⁇ -amino acid is represented by R 3 CH (NH 2 ) COOH.
  • the ⁇ -amino acid is glycine, it is a hydrogen atom, when it is alanine, it is methyl, when it is valine, it is isopropyl, when it is leucine, it is isobutyl, when it is isoleucine, it is sec-butyl, and when it is serine, it is hydroxymethyl.
  • the side chain of the ⁇ -amino acid of R 1 may be combined with the carbon atom to which it is bonded and the nitrogen atom adjacent to the carbon atom to form a ring.
  • the cyclic moiety herein include a proline cyclic moiety (that is, a pyrrolidine ring).
  • the side chain of the ⁇ -amino acid of R 1 does not form a ring with the carbon atom to which it is attached and the nitrogen atom adjacent to the carbon atom.
  • the “ ⁇ -amino acid” of R 1 preferably has an active group such as carboxy group, hydroxy group or mercapto group on the side chain of serine, threonine, cysteine, tyrosine, aspartic acid, glutamic acid, homoserine, homocysteine, penicillamine and the like.
  • the amino acid is selected from glutamic acid and aspartic acid; and particularly preferable is glutamic acid.
  • the number of substitution of a monovalent group consisting of boron clusters or a monovalent group to which it is bonded via a linker structure in R 1 is preferably 1 to 3, and more preferably 1.
  • each R 1 is independently a monovalent group composed of a boron cluster (particularly preferably, a group represented by any one of the formulas (B1) to (B15)) is bonded via a linker structure.
  • R 1 is independently ⁇ - selected from a neutral amino acid and an acidic amino acid, wherein each group represented by the formula (B1) is substituted with a monovalent group bonded via a linker structure. A side chain of an amino acid.
  • R 1 is more preferably the case where the “ ⁇ -amino acid” of R 1 is selected from the group consisting of glutamic acid and aspartic acid, and each independently represents formula (R1a):
  • the peptide compound comprises a bond of the above boron cluster (including a linker structure) and a polyether (eg, “monovalent group composed of polyether”, “c-valent group composed of polyether”).
  • a polyether eg, “monovalent group composed of polyether”, “c-valent group composed of polyether”.
  • it may further have other substituents.
  • one or more substituents may be substituted with the side chain of the amino acid, and among them, the active group of the side chain of the amino acid (for example, carboxy group, hydroxy group and mercapto group) H or It is preferably substituted with OH.
  • the boron cluster may not be introduced due to unreacted, and the linker structure or a fragment structure thereof may remain. Residual unreacted structures may be included.
  • the other substituents include structures obtained by inactivating unreacted structures by methods known to those skilled in the art, structures formed by side reactions, and undeprotected groups that were not removed by deprotection reactions. Structures and the like can also be included.
  • the terminal of the linker structure or its fragment structure left unreacted can take a wide variety of structures and is not limited to a specific substituent.
  • the above-mentioned other substituents may include modifying groups known to those skilled in the art used in peptide chemistry.
  • a 1 ′ represents an unreacted functional group in a known coupling reaction, a group inactivated by a known method, a group generated by a side reaction thereof, or a group containing an undeprotected group.
  • g ′ represents an integer of 0 to 19, and other symbols are as defined above.
  • the group represented by these is mentioned.
  • the unreacted functional group include a mercapto group, a pyridin-2-yldisulfanyl group, a carboxy group or an active group thereof (for example, an active ester such as 4-nitrophenol ester and pentafluorophenol ester, and an acid chloride).
  • Non-acid halides Non-acid halides), amino groups, azide (N 3 ) groups for cyclization reactions by click chemistry, cyclooctyne groups, ethynyl (HC ⁇ C) groups, and the like.
  • examples of the other substituents include, but are not limited to, a substituent selected from the group of substituent A consisting of the following (1) to (5); group: (1) 1 to 5 substituents selected from the following substituents (a) to (l), which may be substituted on alkyl, alkenyl, alkynyl or aryl, respectively, AA): (A) an alkyl group having 1 to 6 carbon atoms, (B) an alkenyl group having 2 to 6 carbon atoms, (C) an alkynyl group having 2 to 6 carbon atoms, (D) an aryl having 6 to 14 carbon atoms (E) an alkylcarbonyl group having 2 to 7 carbon atoms, (F) a carboxy group optionally substituted with an alkyl group having 1 to 6 carbon atoms, and (G) an alkenyl group having 2 to 6 carbon atoms.
  • substituents selected from the group of substituent A consisting of the following (1) to (5); group: (1) 1 to
  • An amino group which may be substituted (T) an amino group which may be substituted with an alkynyl group having 2 to 6 carbon atoms, and (U) an amino group which may be substituted with an aryl group having 6 to 14 carbon atoms (V) substituted with an alkyl group having 1 to 6 carbon atoms
  • An optional mercapto group (W) a mercapto group optionally substituted with an alkenyl group having 2 to 6 carbon atoms, (X) a mercapto group optionally substituted with an alkynyl group having 2 to 6 carbon atoms, (Y) a mercapto group optionally substituted with an aryl group having 6 to 14 carbon atoms, (Z) an alkylsulfonyl group having 1 to 6 carbon atoms, and (AA) an alkyl group having 1 to 6 carbon atoms.
  • An optionally substituted sulfo group (A) an optionally substituted aryl group having 6 to 14 carbon atoms, (b) a formyl group, (c) an optionally substituted alkylcarbonyl group having 2 to 7 carbon atoms, (d) a substituted group An optionally substituted alkyl group having 1 to 6 carbon atoms, an optionally substituted alkenyl group having 2 to 6 carbon atoms, an optionally substituted alkynyl group having 2 to 6 carbon atoms, and a substituted group.
  • Substituent group A preferably has 1 to 5 substituents selected from the above substituents (a) to (l), and each of the above substituents (A), ( E), (F), (J), (Z), (R), (V), (Z) and (AA).
  • substituent group A 1 to 5 substituents selected from the following substituents (c ′) to (l ′) are substituted with the above substituents (A ), (E), (F), (J), (Z), (R), (V), (Z) and (AA); (C ′) an optionally substituted alkylcarbonyl group having 2 to 7 carbon atoms, (d ′) an optionally substituted carboxy group having 1 to 6 carbon atoms, (E ′) an optionally substituted carbamoyl group having 1 to 6 carbon atoms, (f ′) an optionally substituted alkyl group having 1 to 6 carbon atoms A hydroxy group which may be substituted, (g ′) a mercapto group which may be substituted with an alkyl group having 1 to 6 carbon atoms which may be substituted, and (h ′) a carbon number which may be substituted.
  • the substituent group A is more preferably the above substituent (R) which may be substituted on the alkyl with 1 to 5 substituents selected from the above substituents (c ′) to (l ′). .
  • substituent of the “alkylsulfonyl group having 1 to 6 carbon atoms” include the substituent group B consisting of the following (1) to (15); Substituent group B: (1) an alkyl group having 1 to 6 carbon atoms, (2) an alkenyl group having 2 to 6 carbon atoms, (3) an alkynyl group having 2 to 6 carbon atoms, and (4) an aryl having 6 to 14 carbon atoms.
  • alkenyl group having 2 to 6 carbon atoms refers to a straight, branched or cyclic partially unsaturated hydrocarbon having 2 to 6 carbon atoms having at least one double bond.
  • alkynyl group having 2 to 6 carbon atoms means a linear, branched or cyclic partially unsaturated hydrocarbon group having 2 to 6 carbon atoms and having at least one triple bond.
  • alkylcarbonyl group having 2 to 7 carbon atoms means a group to which the above “alkyl group having 1 to 6 carbon atoms” is bonded via carbonyl, such as acetyl, propionyl, butyryl. , Isobutyryl, sec-butylcarbonyl, tert-butylcarbonyl, pentanoyl, hexanoyl and the like.
  • alkylsulfonyl group having 1 to 6 carbon atoms means a group to which the above “alkyl group having 1 to 6 carbon atoms” is bonded via sulfonyl, for example, methylsulfonyl, ethylsulfonyl Propylsulfonyl, isopropylsulfonyl, butylsulfonyl, isobutylsulfonyl, sec-butylsulfonyl, tert-butylsulfonyl, pentylsulfonyl and the like.
  • sulfonyl for example, methylsulfonyl, ethylsulfonyl Propylsulfonyl, isopropylsulfonyl, butylsulfonyl, isobutylsulfonyl, sec-butylsulfonyl, tert-butyl
  • the molecular weight of the peptide compound of the present invention or a salt thereof is preferably less than 100,000, more preferably less than 80,000, still more preferably less than 70,000, and less than 60,000. Even more preferred.
  • the peptide compound of the present invention or a salt thereof is a compound represented by the formula (Ia) or the formula (Ib).
  • R 1 has the same meaning as described above, and each independently represents a side chain of an ⁇ -amino acid substituted with a monovalent group consisting of a boron cluster or a monovalent group to which it is bonded via a linker structure.
  • the “monovalent group consisting of boron clusters”, “linker structure”, and “side chain of ⁇ -amino acid” have the same meaning as described above.
  • R 2 independently represents a side chain of an ⁇ -amino acid selected from a neutral amino acid and an acidic amino acid, which may have a substituent.
  • the “ ⁇ -amino acid” of R 2 is preferably substituted with carboxy in the side chain of serine, threonine, cysteine, tyrosine, aspartic acid, glutamic acid, homoserine, homocysteine, penicillamine, etc.
  • An ⁇ -amino acid having an active group such as a group, a hydroxy group, a mercapto group, etc., and an unsubstituted ⁇ -amino acid when unsubstituted.
  • the “ ⁇ -amino acid” of R 2 is more preferably a side chain such as serine, threonine, cysteine, tyrosine, aspartic acid, glutamic acid, homoserine, homocysteine, penicillamine, etc., when substituted.
  • the “ ⁇ -amino acid” of R 2 is preferably a serine, threonine, cysteine, tyrosine, aspartic acid, glutamic acid, homoserine, homocysteine, penicillamine, etc. in the side chain such as carboxy group, hydroxy group, mercapto
  • the side chain of the ⁇ -amino acid of R 2 may be combined with the carbon atom to which it is attached and the nitrogen atom adjacent to the carbon atom to form a ring.
  • the cyclic moiety herein include a proline cyclic moiety (that is, a pyrrolidine ring).
  • the side chain of the ⁇ -amino acid of R 2 does not form a ring with the carbon atom to which it is attached and the nitrogen atom adjacent to the carbon atom.
  • Examples of the substituent on the side chain of the ⁇ -amino acid of R 2 include those similar to the above-mentioned other substituents in addition to the monovalent group consisting of polyether. In one embodiment, the substituent at R 2 is absent.
  • the side chain of the ⁇ -amino acid may be substituted with any H or OH on the carbon atom, nitrogen atom or sulfur atom, for example, preferably H and OH of the amino acid active group.
  • a group selected from OH for example, OH of a carboxy group and H of a hydroxy group and a mercapto group
  • R 2 Specific examples of the “monovalent group consisting of polyether” in R 2 include groups represented by any of the above formulas (P1) to (P3). Among them, a group represented by the above formula (P1) is preferable.
  • the ⁇ -amino acid residue having R 1 and the ⁇ -amino acid residue having R 2 are arbitrarily copolymerized, and even if they are randomly copolymerized, they are block-copolymerized. Alternatively, alternating copolymerization may be performed.
  • all of the “ ⁇ -amino acid” of R 1 and the “ ⁇ -amino acid” of R 2 are the same ⁇ -amino acid.
  • X 1 and Y 2 when c is 1 are each independently (1) a hydrogen atom, (2) an alkyl group having 1 to 6 carbon atoms, and (3) an alkylcarbonyl having 2 to 7 carbon atoms.
  • a monovalent group consisting of a group or (4) a polyether;
  • X 1 is preferably a hydrogen atom or a monovalent group consisting of a polyether, and more preferably a hydrogen atom.
  • Y 2 in the case where c is 1 is preferably a hydrogen atom or a monovalent group consisting of a polyether, and more preferably a monovalent group consisting of a polyether.
  • X 2 and Y 1 when c is 1 are each independently (1) a hydroxy group which may be substituted with an alkyl group having 1 to 6 carbon atoms, and (2) a carbon number of 1 to 6
  • X 2 is preferably a hydroxy group which may be substituted with an alkyl group having 1 to 6 carbon atoms, or an amino group which may be substituted with an alkyl group having 1 to 6 carbon atoms.
  • Y 1 is preferably a monovalent group made of a polyether.
  • Y 1 and Y 2 each represent a c-valent group made of a polyether.
  • X 1 , X 2 , Y 1 or Y 2 is a monovalent group consisting of a polyether or a c-valent group consisting of a polyether
  • these are an amino group (NH of an amino acid residue) and a carboxy group (amino acid residue).
  • bonds that can be formed with the group (CO) include the same bonds as those described above as the bonds that the boron cluster (or linker structure) can form.
  • Specific examples of the “monovalent group consisting of polyether” in X 1 , X 2 , Y 1 and Y 2 include groups represented by any of the above formulas (P1) to (P3). Among them, a group represented by the above formula (P1) is preferable.
  • Specific examples of the “c-valent group made of polyether” in Y 1 and Y 2 include a group represented by the above formula (P4) or (P5) when c is 2.
  • the total of a and b is an integer of 2 or more, preferably an integer of 3 or more, more preferably an integer of 5 or more, and further preferably an integer of 10 or more.
  • the sum of a and b is preferably an integer of 200 or less, more preferably an integer of 150 or less, and still more preferably an integer of 100 or less.
  • the ratio of a to the sum of a and b is preferably 0.1 or more, more preferably 0.2 or more, and still more preferably 0.3 or more.
  • formulas (Ia) and (Ib) are represented by formulas (IA) and (IB), respectively:
  • the peptide compound of the present invention may be a salt.
  • the salt include metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic amino acids or acidic amino acids, and the like.
  • the metal salt include alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt and magnesium salt; aluminum salt and the like.
  • the ammonium salt include tetramethylammonium salt, tetrabutylammonium salt, and monovalent or polyvalent ammonium salt of polyamine (eg, 1,4-diaminobutane, spermine, spermidine).
  • the salt with an organic base include, for example, trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N, N′-dibenzyl.
  • examples thereof include salts with amines such as ethylenediamine and polyamine (eg, 1,4-diaminobutane, spermine, spermidine).
  • Preferable examples of the salt with inorganic acid include salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid and the like.
  • the salt with organic acid include, for example, formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzene And salts with sulfonic acid, p-toluenesulfonic acid and the like.
  • Preferable examples of salts with basic amino acids include salts with arginine, lysine, ornithine and the like, and preferable examples of salts with acidic amino acids include salts with aspartic acid, glutamic acid and the like. Is mentioned. Among them, the salt of the peptide compound of the present invention is preferably a pharmaceutically acceptable salt.
  • the peptide compound of the present invention or a salt thereof (hereinafter referred to as the compound of the present invention) can be used for boron neutron capture therapy for tumor diseases.
  • tumor disease includes malignant melanoma, renal cancer, prostate cancer, breast cancer, lung cancer, pancreatic cancer, colon cancer, hepatocellular carcinoma, biliary tract cancer, stomach cancer, ovarian cancer, esophageal cancer, urinary tract Skin cancer, colon cancer, bone cancer, skin cancer (eg malignant skin cancer), head and neck cancer, uterine cancer, rectal cancer, anal cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer , Vaginal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, childhood solid cancer, bladder cancer, malignant pleural mesothelioma, brain tumor (eg malignant brain tumor) And central nervous system tumors.
  • malignant melanoma renal cancer, prostate cancer, breast cancer, lung cancer, pancreatic cancer, colon cancer, hepatocellular carcinoma, biliary tract cancer,
  • Boron neutron capture therapy using a compound of the present invention can be performed, for example, on a mammal (eg, a human) suffering from a tumor disease by applying a drug containing the compound of the present invention by any suitable route of administration such that the compound accumulates at the target site.
  • a mammal eg, a human
  • the compound of the present invention is preferably accumulated selectively in a tumor.
  • Formulations containing the compound may be administered at once or sequentially. Administration of the formulation may be repeated as necessary.
  • the site is irradiated with an effective amount of a low energy neutron beam such as a thermal neutron beam or an epithermal neutron beam.
  • a low energy neutron beam such as a thermal neutron beam or an epithermal neutron beam.
  • the site may be irradiated through the skin, or the site may be irradiated completely or partially exposed prior to irradiation. Moreover, you may irradiate from several directions simultaneously. Administration of the compound of the present invention and subsequent irradiation with thermal neutron rays or epithermal neutron rays may be repeated as necessary. For example, you may irradiate several times at intervals of about several months.
  • the total number of times of irradiation is preferably 1 to 10 times, more preferably 1 to 5 times.
  • the administration route of the compound of the present invention is preferably intravenous administration, intraarterial administration, intramuscular administration, subcutaneous administration, intradermal administration, intraspinal administration, intraperitoneal administration.
  • the compound of the present invention is a mammal suffering from a tumor disease within 72 hours (preferably 5 minutes to 48 hours, more preferably 30 minutes to 30 hours before) of irradiation with thermal neutrons or epithermal neutrons. Can be administered to animals. Moreover, you may administer during the irradiation of a thermal neutron beam or an epithermal neutron beam as needed.
  • a typical dose of the compound of the present invention is preferably in the range of 0.01 mg to 50 g / kg of body weight per irradiation of thermal neutron beam or epithermal neutron beam.
  • boron cluster it is preferably in the range of 0.005 mg to 25 g / kg of body weight per irradiation of thermal neutron beam or epithermal neutron beam.
  • the irradiation time of one thermal neutron beam or epithermal neutron beam is preferably 1 minute to 5 hours, more preferably 10 minutes to 2 hours.
  • the irradiation amount of the thermal neutron beam or the epithermal neutron beam is not particularly limited as long as it is a general irradiation amount used in neutron capture therapy.
  • Boron neutron capture therapy using the compounds of the present invention may be applied as a single treatment or may be applied with conventional surgery or chemotherapy. If desired, after removing the tumor to the extent possible surgically, the remaining tumor can be destroyed by boron neutron capture therapy using the compounds of the present invention.
  • the compound of the present invention can be used together with other boron compounds (hereinafter referred to as concomitant drugs) such as BPA (p-boronophenylalanine) and BSH (mercaptoundecahydrododecaborate).
  • concomitant drugs such as BPA (p-boronophenylalanine) and BSH (mercaptoundecahydrododecaborate).
  • BPA p-boronophenylalanine
  • BSH mercaptoundecahydrododecaborate
  • the administration time of the compound of the present invention and the concomitant drug is not limited, and the compound of the present invention and the concomitant drug may be administered simultaneously to a mammal suffering from a tumor disease, with a time difference. May be administered.
  • the compound of the present invention and the concomitant drug may be administered in this order, or vice versa.
  • the compound of the present invention contains a pharmaceutically acceptable carrier, excipient, binder, stabilizer, dispersant and the like, together with a pharmaceutically acceptable carrier, an injectable solution, suspension, emulsion, cream. And may be formulated into parenteral dosage forms such as ointments, inhalants, suppositories, and the like.
  • the compound of the present invention is preferably formulated into an injectable solution.
  • Injection solutions can be prepared as solutions of the compounds of the invention in pharmaceutically acceptable solvents. These solutions may also contain stabilizing components and / or buffering components. In addition, the injection solution may be a dry preparation to which an appropriate solvent is added before use.
  • compounds represented by formulas (Ia ′) and (Ib ′) are represented by formulas (IIa) and (IIb).
  • the ⁇ -amino acid side chain of the compound represented (hereinafter referred to as Compound (IIa) or Compound (IIb)) and the compound represented by Formula (B) (hereinafter referred to as Compound (B)) are converted into A 2.
  • a 1 can be coupled by a known condensation reaction, addition reaction and substitution reaction to form A, and can be obtained by bonding directly or via a linker structure.
  • an active group such as a carboxy group, a hydroxy group, an amino group, or a mercapto group is present on the reaction substrate, these functional groups may be protected in advance with a known protecting group.
  • the wavy line portion is a bond or represents a linker structure or a fragment structure thereof, and A 1 is a side chain of an ⁇ -amino acid, a linker group or a functional group in the fragment structure bound thereto, A 2 is a functional group in a boron cluster, a linker structure bonded to the boron cluster, or a fragment structure thereof.
  • A is a part of the side chain of P 2 and ⁇ -amino acid, or a structure corresponding to Q 2 .
  • A includes —O—, —S—, —NH—, —SO 2 —, —CO—, —NHCO—, —CONH—, —OCO—, —COO—, —SCO—, —COS—.
  • A is —SS—
  • one of A 1 and A 2 may be a mercapto group and the other may be a pyridin-2-yldisulfanyl group.
  • Reacting compound (IIa) or compound (IIb) with compound (B) in a solvent for example, alcohols such as methanol and ethanol, sulfoxides such as dimethylsulfoxide (DMSO), or a mixture thereof.
  • a solvent for example, alcohols such as methanol and ethanol, sulfoxides such as dimethylsulfoxide (DMSO), or a mixture thereof.
  • DMSO dimethylsulfoxide
  • the reaction temperature is usually from -80 to 150 ° C.
  • A is —NHCO— or —CONH—
  • one of A 1 and A 2 is a carboxy group or an active group thereof (for example, an active ester such as 4-nitrophenol ester, pentafluorophenol ester, , Acid halide such as acid chloride, etc.) and the other may be an amino group.
  • Compound (IIa) or Compound (IIb) and Compound (B) are mixed with a solvent (for example, sulfoxides such as dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), N, N-dimethylformamide ( Amides such as DMF), or a mixture thereof, if necessary, a condensing agent (for example, a carbodiimide-based condensing agent such as 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide or a salt thereof); 4- (4,6-dimethoxy-1,3,5-triazin-2-yl) -4-methylmorpholinium chloride and the like), an activator (for example, N-hydroxysuccinimide, 1 -Hydroxybenzotriazole (HOBt), 4-dimethylaminopyridine (DMAP), etc.), base (for example, N-methylmol Phosphorus (NMM), by reacting in the
  • A is —SCO— or —COS—
  • one of A 1 and A 2 is a carboxy group or an active base thereof (for example, an active ester such as 4-nitrophenol ester, pentafluorophenol ester, , Acid halide such as acid chloride, etc.) and the other may be a mercapto group.
  • an active ester such as 4-nitrophenol ester, pentafluorophenol ester, , Acid halide such as acid chloride, etc.
  • the other may be a mercapto group.
  • Compound (IIa) or Compound (IIb) and Compound (B) are mixed with a solvent (for example, sulfoxides such as dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), N, N-dimethylformamide ( Amides such as DMF), or a mixture thereof, if necessary, a condensing agent (for example, a carbodiimide-based condensing agent such as 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide or a salt thereof); 4- (4,6-dimethoxy-1,3,5-triazin-2-yl) -4-methylmorpholinium chloride and the like), an activator (for example, N-hydroxysuccinimide, 1 -Hydroxybenzotriazole (HOBt), 4-dimethylaminopyridine (DMAP), etc.), base (for example, N-methylmol Phosphorus (NMM), by reacting in the
  • A is -heterocyclic (1,2,3-triazole)-
  • one of A 1 and A 2 is an azide (N 3 ) group and the other is an ethynyl (HC ⁇ C) group If it is.
  • Compound (IIa) or Compound (IIb) and Compound (B) are mixed with a solvent (for example, amides such as N, N-dimethylformamide (DMF); ketones such as acetone; methanol, ethanol, tert-butanol In water (including buffer); or mixtures thereof) in copper catalysts (eg, copper (I) sulfate, copper (I) bromide, copper (I) iodide, trifluoromethanesulfonic acid) Copper (I) etc.), if necessary, additives (for example, reducing agents such as sodium ascorbate, tris (2-carboxyethyl) phosphine (TCEP), etc.), ligands (for example, tri
  • linker structure or the fragment structure thereof is introduced into the side chain of an ⁇ -amino acid in the same manner as in the construction method of A described above or by a known method. can do.
  • an active group such as a carboxy group, a hydroxy group, an amino group, or a mercapto group
  • these functional groups may be protected in advance with a known protecting group.
  • the linker structure or the fragment structure thereof can be introduced into the boron cluster in the same manner as in the construction method of A described above or by a known method.
  • an active group such as a carboxy group, a hydroxy group, an amino group, or a mercapto group
  • these functional groups may be protected in advance with a known protecting group.
  • a linker structure or a fragment structure thereof can be introduced into the active group in the same manner as in the above construction method A.
  • a method of directly introducing a linker structure or a fragment structure thereof into a boron cluster having no active group is also known, and examples thereof include a method represented by the following scheme.
  • the compound represented by the formula (2) is obtained by converting the boron cluster (1) into a compound represented by the formula (3), an acid (for example, hydrochloric acid (hydrogen chloride), etc.), an additive (for example, sodium tetrafluoroborate). Tetrafluoroborate, etc.), and optionally a solvent (for example, sulfoxides such as dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), N, N-dimethylformamide (DMF) Can be obtained by the reaction in the presence of amides such as The reaction temperature is usually from -80 to 150 ° C.
  • an acid for example, hydrochloric acid (hydrogen chloride), etc.
  • an additive for example, sodium tetrafluoroborate). Tetrafluoroborate, etc.
  • a solvent for example, sulfoxides such as dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), N, N-dimethylformamide
  • the compound represented by the formula (4) is obtained by replacing the compound represented by the formula (2) with a corresponding nucleophile, optionally a base (for example, an organic base such as triethylamine, sodium hydroxide, potassium hydroxide). , Nucleophilic addition reaction in the presence of an inorganic base such as potassium carbonate or cesium fluoride).
  • the reaction temperature is usually from -80 to 150 ° C. At that time, when an active group such as a carboxy group, a hydroxy group, an amino group, or a mercapto group is present on the reaction substrate, these functional groups may be protected in advance with a known protecting group.
  • the reaction temperature is usually from -80 to 150 ° C.
  • the nucleophile corresponding to the introduction group when introducing an azide (—N 3 ) group or an amino (—NH 2 ) group, for example, tetrabutylammonium azide is used, and in particular, when introducing an amino group.
  • a reducing agent such as hydrogen gas
  • a catalyst such as palladium carbon.
  • the reaction temperature is usually from -80 to 150 ° C.
  • HS— (CH 2 ) 2 —NHBoc 2-aminoethylthio (—S— (CH 2 ) 2 —NH 2 ) group
  • HS— (CH 2 ) 2 —NHBoc is used, and after the reaction, the Boc group is converted to an acid
  • deprotection is performed using hydrochloric acid, trifluoroacetic acid, or the like.
  • the reaction temperature is usually from -80 to 150 ° C.
  • the compound represented by the formula (5) is obtained by reacting the compound represented by the formula (2) in the presence of thiourea and a solvent (for example, alcohols such as methanol, ethanol, tert-butanol), Then, it can obtain by processing with a base (for example, sodium methoxide etc.).
  • a solvent for example, alcohols such as methanol, ethanol, tert-butanol
  • the reaction temperature is usually from -80 to 150 ° C.
  • the compounds represented by the formulas (IIa ′) and (IIb ′) (hereinafter referred to as the compound (IIa ′) and the compound (IIb) before introducing the linker structure or the fragment structure thereof.
  • a commercially available product may be used as it is, or it may be synthesized using a known peptide elongation method.
  • a peptide elongation reaction is performed from the C-terminus to the N-terminus by a known peptide elongation method.
  • active groups such as carboxy group, hydroxy group, amino group, mercapto group are present in the side chain of ⁇ -amino acid or reaction substrate, these functional groups are protected with known protecting groups in advance. You may keep it.
  • Examples of peptide elongation methods include activated ester method, mixed acid anhydride method, C-terminal activation method such as azide method, coupling method such as carbodiimide, N-carboxylic acid anhydride (NCA) method, redox method Enzyme method, solid phase synthesis method and the like.
  • the degree of polymerization in the chemical structural formula, title, and text indicates an approximate average degree of polymerization.
  • the polymerization degree n of the polyether in the chemical structural formula has an independent value for each synthesis example and each example.
  • 10 B- indicates a compound having an isotope ratio of 10 B of 98% or more
  • natural B- indicates a natural isotope ratio ( 11 B and 19 of 80.1%). 9% of 10 B).
  • ⁇ -benzyl L-glutamate N-carboxylic anhydride (BLG-NCA, 526 mg, 2.0 mmol) was dissolved in DMF (26 ml) and then dissolved in DMF (30 ml).
  • the compound (300 mg) obtained in Synthesis Example 1 was dialyzed in 0.01N hydrochloric acid to ion-exchange a polyglutamic acid counter cation from a sodium cation to a proton, and then lyophilized. .
  • the obtained compound, EDC (147.4 mg, 1.1 eq), N-hydroxysuccinimide (NHS) (88.5 mg, 1.1 eq) was stirred in DMSO (10 ml) for 30 minutes.
  • 2- (Pyridinyldisulfanyl) ethanamine hydrochloride (156.5 mg, 1.0 eq) was added, and the mixture was further stirred for 24 hours.
  • the compound (10 mg) obtained in Synthesis Example 2 was dissolved in DMSO (10 ml), and 10 B-BSH (4.2 mg, thiolated in a block copolymer) dissolved in methanol (2 ml). After mixing with 2 eq) for the functional group and reacting overnight at room temperature, the reaction solution was dialyzed in a thin aqueous sodium hydroxide solution and freeze-dried to obtain the title compound.
  • the title compound was synthesized by condensing the compound obtained in Synthesis Example 4 with 10 B-BSH in the same manner as in Example 1. In the same manner as in Example 1, about 48 10 B-BSH was introduced per molecule of polymer, and the introduced amount of 10 B-BSH per polymer molecule was 27.6% (w / w). Decided to.
  • the title compound was synthesized by condensing the compound obtained in Synthesis Example 6 with 10 B-BSH in the same manner as in Example 1. In the same manner as in Example 1, about 20 10 B-BSH was introduced per molecule of polymer, and the introduced amount of 10 B-BSH per polymer molecule was 13.8% (w / w). Decided to.
  • the molecular weight distribution was calculated to be 1.15 by the same method as in Synthesis Example 1.
  • Synthesis example 8 Synthesis of thiolated polyglutamic acid (40mer) (P (Glu-Thiolate))
  • the title compound was synthesized by condensing the compound obtained in Synthesis Example 8 with 10 B-BSH in the same manner as in Example 1. In the same manner as in Example 1, about 19 10 B-BSH was introduced per molecule of polymer, and the introduced amount of 10 B-BSH per molecule was 38.6% (w / w). Decided to.
  • the molecular weight distribution was calculated to be 1.15 by the same method as in Synthesis Example 1.
  • the title compound was synthesized by condensing the compound obtained in Synthesis Example 10 with 10 B-BSH in the same manner as in Example 1. In the same manner as in Example 1, about 41 10 B-BSH was introduced per molecule of polymer, and the introduced amount of 10 B-BSH per polymer molecule was 38.0% (w / w). Decided to.
  • ⁇ -Benzyl L-aspartate N-carboxylic anhydride (BLA-NCA) (200 mg, 0.803 mmol) was added dropwise and stirred at 40 ° C. for 16 hours.
  • the reaction solution was re-precipitated dropwise in hexane / ethyl acetate (3/2 (v / v), 50 ml) to give the title compound (392 mg, 0.0295 mmol, yield 91%) as white.
  • Mw / Mn The molecular weight distribution (Mw / Mn) was 1.06 by size exclusion chromatography.
  • BLG-NCA 200 mg, 0.760 mmol was added dropwise and stirred at room temperature for 16 hours.
  • the reaction mixture was re-precipitated dropwise in hexane / ethyl acetate (3/2 (v / v), 50 ml) under ice cooling to give the title compound (384 mg, 0.0248 mmol, yield 82%). Obtained as a white powder.
  • Mw / Mn The molecular weight distribution (Mw / Mn) was 1.35 by size exclusion chromatography.
  • -NCA (1.70 g, 6.46 mmol) was added dropwise and stirred at room temperature for 16 hours. Under ice-cooling, the reaction mixture was added dropwise to hexane / ethyl acetate (3/2 (v / v), 750 ml) for reprecipitation to give the title compound (3.70 g, 0.244 mmol, yield). 95%) was obtained as a white powder.
  • Natural B-dodecahydrododecaborate-2 tetrabutylammonium salt (7.50 g, 12.0 mmol) was dissolved in 1,4-dioxane (400 ml), and NaBF 4 (6.57 g, 59.8) was dissolved. mmol), 1,4-dioxane solution (5.99 ml, 23.9 mmol) in 4N hydrogen chloride was added, followed by heating under reflux for 5 hours. The reaction mixture was concentrated under reduced pressure, and washed with slurry with isopropanol to give the title compound (4.17 g, 8.85 mmol, yield 74%) as a white powder.
  • the obtained compound was dissolved in methanol (57.5 ml), and after substitution with argon, 5% Pd / C (2.87 g, of which moisture was 52.5%) was added. Under a hydrogen atmosphere, Stir for 5 hours. After filtration through Celite (registered trademark), the filtrate was concentrated under reduced pressure to obtain the title compound (1.91 g, 2.63 mmol, yield 91%) as a white powder.
  • Cysteamine hydrochloride (530 mg, 4.67 mmol) was dissolved in dichloromethane (10 ml), and then Boc 2 O (1.43 g, 6.57 mmol) dissolved in dichloromethane (5.0 ml) was added. It was dripped. Subsequently, triethylamine (754 ⁇ l, 5.59 mmol) was added, and the mixture was stirred at room temperature for 24 hours. Washing with saturated multistory water, water, and 15% saline in order, drying over anhydrous magnesium sulfate, filtering, and concentrating the filtrate under reduced pressure gave a crude product of protected cysteamine (1.03 g). It was.
  • N-protected aminohexanoic acid (1.2 g, 4.54 mmol) was dissolved in acetonitrile (10 ml), 4-nitrophenyl chloroformate (1.47 ml, 7.29 mmol), triethylamine. (0.944 ml, 7.09 mmol) and DMAP (82.0 mg, 0.668 mmol) were added and stirred at 4 ° C. for 45 minutes.
  • the reaction mixture was concentrated and purified by silica gel column chromatography to give the title compound (632 mg, 1.64 mmol, yield 36%) as a pale yellow oil.
  • the reaction solution was dialyzed 3 times in 150 mM saline, then dialyzed 3 times in deionized water (fractionated molecular weight 3500 Da), and lyophilized to obtain the title compound (266 mg, yield 96%).
  • the introduction rate of 10 B-BSH was determined from the degree of polymerization of boron cluster protons (-B H ; ⁇ (ppm): 0.5-2.0) and polyamino acid by 1 H-NMR analysis (D 2 O). , 51%, and 49% by quantification of the amount of 10 B by ICP-MS analysis. This corresponds to the introduction of about 20 10 B-BSH per molecule of polymer.
  • Test example 1 Cytotoxicity test against HUVEC The compounds of Examples 1 to 3 and Comparative Examples 1 and 2 and 10 B-BSH were examined for cytotoxicity against HUVEC (human umbilical vein endothelial cells) by CCK-8 assay. HUVECs were suspended in an endothelial cell growth medium kit, seeded in a 96-well plate so that there were 5000 cells in each well, and cultured at 37 ° C. in the presence of 5% carbon dioxide for 24 hours. Subsequently, any one of the compounds of Examples 1 to 3 and Comparative Examples 1 and 2 and 10 B-BSH was added, and the cells were further cultured at various concentrations for 72 hours.
  • HUVEC human umbilical vein endothelial cells
  • Test example 2 Cytotoxicity test for C26 cancer cells
  • the compounds of Examples 1 to 3 and Comparative Examples 1 and 2, and 10 B-BSH were examined for cytotoxicity to C26 cancer cells in the same manner as in Test Example 1. As shown in FIG. 2, cytotoxicity against C26 cancer cells was not observed for the compounds of Examples 1 to 3, Comparative Examples 1 and 2, and 10 B-BSH.
  • Test example 3 Intracellular uptake test using C26 cancer cells The amount of intracellular uptake of the compounds of Examples 1 to 3 and Comparative Examples 1 and 2 and 10 B-BSH was examined.
  • 10 6 C26 cancer cells were seeded with 2 mL of Dulbecco's modified Eagle medium (DMEM medium) and cultured for 24 hours.
  • DMEM medium Dulbecco's modified Eagle medium
  • the compound of Example 1, the compound of Example 2, the compound of Example 3, the compound of Comparative Example 1, the compound of Comparative Example 2, and 10 B-BSH were each converted to 10 B-BSH to 100 ⁇ g / mL. It added so that it might become.
  • each cell was washed 3 times with PBS (phosphate buffer), and treated with trypsin to collect the cells. The number of collected cells was counted, ashed with 90% nitric acid, finally diluted with 1% nitric acid, and the amount of boron was quantified by ICP-MS analysis.
  • PBS phosphate buffer
  • the peptide compounds with 10 B-BSH of Examples 1 to 3 and Comparative Examples 1 and 2 had significantly higher intracellular uptake than 10 B-BSH alone. . Furthermore, the amount of cellular uptake of the compounds of Examples 1 to 3 having PEG was significantly higher than the compounds of Comparative Examples 1 and 2 having no PEG.
  • Test example 4 Intracellular uptake test using C26 cancer cells It was confirmed that the compound of Example 1 entered the cells. First, the compound of Example 1 was fluorescently labeled by condensation with Alexa488-NHS ester. Subsequently, the obtained compound was added to C26 cancer cells as 10 B-BSH so as to be 20 ⁇ g / mL, and subsequently observed with a confocal laser microscope (CLSM). As shown in FIG. 4, it was confirmed that the cancer cells entered from 1 hour after the addition.
  • CLSM confocal laser microscope
  • Test Example 5 Intracellular uptake test using CT26 cancer cells It was confirmed that boron clusters were incorporated more into cancer cells by introducing them into peptides containing PEG.
  • CT26 carcinoma cells murine colon carcinoma cells, ATCC CRL-2638
  • RPMI1640 medium RPMI1640 medium
  • the cells were washed twice with PBS (0.5 ml), and the cells detached from the plate using trypsin were collected in a 14 ml round tube. After adding 10% nitric acid (0.9 ml) and 60% nitric acid (0.4 ml) and allowing to stand for 30 minutes, ashing was carried out by heating at 50 ° C. for 30 minutes and further at 90 ° C. for 2 hours. . This solution was filtered, diluted 7-fold with deionized water, and the amount of boron in cancer cells was quantified by ICP-MS. As shown in FIG. 5, it was confirmed that the compound of Example 7 was more than twice as much as the counter cation of the compound of Synthesis Example 21 compared to the compound of 2 cesium cation and entered the cancer cell.
  • Test Example 6 Blood retention and tumor accumulation 10 B-BSH and the compounds of Examples 1 to 3 and Comparative Examples 1 and 2 were examined for blood retention and tumor accumulation.
  • the compounds of Examples 1 to 3 and Comparative Examples 1 and 2 and 10 B-BSH were adjusted to a dose of 50 mg / kg in terms of 10 B-BSH.
  • a 0.2 mL volume of physiological saline solution of each compound was intravenously administered, and blood and tumor were collected over time. The blood was heparinized and plasma was obtained by centrifugation.
  • Plasma and tumor samples were treated with hydrochloric acid / 60% nitric acid (3/1) mixed solution or 90% aqueous nitric acid solution to decompose the tissue, diluted with 1% nitric acid, filtered, and then ICP-MS 10 B amount was measured.
  • the compounds of Examples 1 to 3 and Comparative Examples 1 and 2 showed longer retention in blood than 10 B-BSH alone, and the compounds of Examples 1 to 3 having PEG. Showed longer retention in blood than the compounds of Comparative Examples 1 and 2 without PEG.
  • the compounds of Examples 1 to 3 and Comparative Examples 1 and 2 accumulate 10 B-BSH in the tumor 6 to 10 times more than 10 B-BSH alone, and further PEG It was found that the compounds of Examples 1 to 3 having a higher concentration of 10 B-BSH accumulated in the tumor and a longer amount of accumulation than the compounds of Comparative Examples 1 and 2 having no PEG. .
  • Test Example 7 Tumor Accumulation Regarding natural B-BSH and the compound of Example 7, physiological saline solution of each compound was intravenously administered to mice, and tumor accumulation was examined.
  • the amount of the compound of Example 7 was 10 mg / kg and 30 mg / kg in terms of natural B-BSH, and natural B-BSH was added.
  • Each 0.2 mL amount was intravenously administered so that the amount was 100 mg / kg, and tumors were collected over time.
  • the collected tumor samples were heated in a 60% aqueous nitric acid solution at 50 ° C. for 1 hour and then at 90 ° C. for 2 hours to decompose the tissue. After filtration, it was diluted 20 times with pure water, and the amount of natural B was measured by ICP-MS.
  • the compound of Example 7 accumulates boron clusters in the tumor 3 to 5 times more than natural B-BSH alone, and more over time, and maintains a high accumulation amount. I found that the time was long.
  • Test Example 8 Anti-cancer effect
  • the compound of Example 1 or 10 B-BSH was added to each compound so as to be 100 mg / kg in terms of 10 B-BSH.
  • a physiological saline solution (0.2 mL) was intravenously administered. After 24 hours, the mice were locally irradiated with a thermal neutron beam in an amount of 1.6-2.2 ⁇ 10 12 neutron / cm 2 for 1 hour to examine the anticancer effect.
  • V (axb 2 ) / 2 (a and b are the major axis and minor axis when the tumor was measured with a caliper, respectively).
  • BALB / c mice transplanted subcutaneously with C26 cancer cells were treated with 0.2 mL of the compound of Example 1 or 10 B-BSH so that the concentration was 100 mg / kg in terms of 10 B-BSH.
  • the amount of the tumor was administered intravenously, and the size of the tumor was calculated in the same manner without irradiation with thermal neutrons.
  • PBS phosphate buffer solution
  • Tumor size was calculated.
  • the body weight of the mouse used in this experiment was measured as an indicator of the systemic toxicity of the mouse.
  • the mouse of Example 1 or 10 B-BSH alone administered intravenously In mice that were irradiated with neutrons and mice that were not irradiated, there was no apparent weight suppression, and the compounds of the present invention and boron neutron capture therapy using the compounds of the present invention have high safety. all right.
  • Test Example 9 Tumor penetrability A comparison of tumor penetrability using tumor-bearing mice was examined for the liposome preparation Doxil (registered trademark) carrying the compound of Example 1 and the anticancer drug doxorubicin approved as a cancer therapeutic agent. It was. Human pancreatic adenocarcinoma BxPC3 cells, or C26 cancer cell to BALB / c nude mice were implanted subcutaneously, fluorescence 10 mg / kg weight at 10 B terms, the compound of Example 1 with Alexa647 (R) Both the labeled compound and 10 mg / kg Doxil® in terms of doxorubicin were administered intravenously.
  • Doxil registered trademark
  • tumors were excised, sampled and cut to a thickness of 10 ⁇ m with a microtome.
  • the tumor sections were imaged with a confocal scanning microscope LSM780, and the images were normalized in the same way as the background fluorescence intensity.
  • the tumor was similarly cut to a thickness of 5 ⁇ m for hematoxylin-eosin staining, and images were observed with an integrated fluorescence microscope BZ-X700.
  • both the compound obtained by fluorescently labeling the compound of Example 1 with Alexa 647 (registered trademark) and Doxil (registered trademark) were similarly distributed in the polycytic C26 tumor.
  • Doxil registered trademark
  • the compound of Example 1 fluorescently labeled with Alexa 647 (registered trademark) is uniformly distributed in the tumor. did. From these results, it was found that the compound of the present invention has higher tumor penetration than nanoparticles and can deliver boron to the entire cancer cells.
  • the compound of the present invention exhibits excellent cancer accumulation due to the EPR effect and high permeability in tumor tissue due to non-nanoparticulation, it is efficiently taken up into cancer cells, so boron neutron capture therapy It is useful as a pharmaceutical.

Abstract

The purpose of the present invention is to provide a novel boron-containing compound which exhibits excellent accumulation in cancers due to the EPR effect, which does not take nanoparticle form and therefore exhibits high penetration into tumor tissue, and which is efficiently taken up into cancer cells. The present invention relates to a peptide compound having a boron cluster and a polyether, and having as a constituent amino acid an amino acid selected from neutral amino acids and acidic amino acids, or salts of such peptide compound.

Description

ホウ素クラスター結合ペプチド化合物Boron cluster-binding peptide compound
　ホウ素中性子捕捉療法に有用な新規含ホウ素化合物に関し、特に、ホウ素クラスターを導入したペプチド化合物に関する。 It relates to novel boron-containing compounds useful for boron neutron capture therapy, and in particular, to peptide compounds into which boron clusters are introduced.
　中性子捕捉療法（ＮＣＴ）は、外科手術が困難な固形がんに対して優れた治療効果を示し、且つ患者に負担の少ない低侵襲治療法として注目されている。ＮＣＴは、１０Ｂ、１５７Ｇｄ等のような、中性子との反応断面積が大きい元素を含有する分子を体内に投与した後に、患部に対して低エネルギーの熱中性子線或いは熱外中性子線を照射し、核反応によって発生する細胞障害性の放射線（α線等）により、がん細胞を死滅させる治療法である。中でも、ホウ素化合物を用いるホウ素中性子捕捉療法（ＢＮＣＴ）において、臨床応用されている化合物としては、例えばホウ素クラスターのＢＳＨ（メルカプトウンデカハイドロドデカボレート）が知られている。 Neutron capture therapy (NCT) has attracted attention as a minimally invasive treatment method that exhibits an excellent therapeutic effect on solid cancers that are difficult to perform surgery and has less burden on patients. NCT irradiates the affected area with low-energy thermal neutrons or epithermal neutrons after administering molecules containing elements with large reaction cross sections with neutrons such as 10 B and 157 Gd into the body. However, it is a treatment method that kills cancer cells by cytotoxic radiation (α rays or the like) generated by a nuclear reaction. Among them, as a compound applied clinically in boron neutron capture therapy (BNCT) using a boron compound, for example, BSH (mercaptoundecahydrodedecaborate) of a boron cluster is known.
　しかしながら、ＢＳＨは速やかに***され、血中滞留性や腫瘍滞留性に乏しく、現状では非常に高濃度の化合物を体内に投与しながら、患部に熱中性子線或いは熱外中性子線を照射する方法が取られており、この方法では、１０Ｂの腫瘍内濃度が十分ではないためにがん細胞を根治できない場合があること、正常組織への傷害が起こりうること、さらには熱中性子線或いは熱外中性子線のピンポイント照射が必要なために適用できる疾患が限定される等の問題があった。 However, BSH is rapidly excreted and has poor blood retention and tumor retention. Currently, there is a method of irradiating the affected area with a thermal neutron beam or an epithermal neutron beam while administering a very high concentration compound into the body. In this method, cancer cells may not be able to be completely cured due to insufficient concentration of 10 B in the tumor, damage to normal tissue may occur, thermal neutron radiation or There is a problem that applicable diseases are limited because pinpoint irradiation with neutron beams is necessary.
　これに対し、これまで、腫瘍内の１０Ｂ濃度を高めることを目的として、抗体／ホウ素化合物コンジュゲート、カチオン性ポリマー／ホウ素化合物コンジュゲート、Ｃｅｌｌ　Ｍｅｍｂｒａｎｅ　Ｐｅｎｅｔｒａｔｉｎｇ　Ｐｅｐｔｉｄｅ　（ＣＰＰ）／ホウ素化合物コンジュゲート、ホウ素化合物封入リポソーム、高分子ミセル等の高濃度のホウ素化合物をがん組織に送達できるさまざまなドラッグデリバリーシステム（ＤＤＳ）が開発されてきた［Ｍ．Ｊ．Ｌｕｄｅｒｅｒ　ｅｔ　ａｌ．，Ｐｈａｒｍ．Ｒｅｓ．，３２：２８２４－２８３６（２０１５）］。 In contrast, for the purpose of increasing the 10 B concentration in the tumor, antibody / boron compound conjugate, cationic polymer / boron compound conjugate, Cell Membrane Penetrating Peptide (CPP) / boron compound conjugate, boron Various drug delivery systems (DDS) capable of delivering high concentration boron compounds such as compound-encapsulated liposomes and polymer micelles to cancer tissues have been developed [M. J. et al. Luderer et al. Pharm. Res. , 32: 2824-2836 (2015)].
　特に、リポソーム、高分子ミセルのようなナノ粒子の場合には、血中滞留性を上げ、Ｅｎｈａｎｃｅｄ　Ｐｅｒｍｅａｂｉｌｉｔｙ　ａｎｄ　Ｒｅｔｅｎｔｉｏｎ　（ＥＰＲ）効果（がん組織では腫瘍血管の透過性の亢進と未発達なリンパ系の構築によって高分子物質が集積しやすい環境が形成されている効果）によって、がん組織に選択的に集積し、さらに腫瘍における長期滞留性によって、低分子量のＢＳＨと比較して高いがん組織／血中１０Ｂ濃度比が達成された後に、熱中性子線或いは熱外中性子線の照射を行うことができるため、広く研究されてきた。 In particular, in the case of nanoparticles such as liposomes and polymer micelles, the retention in blood is increased, and enhanced permeability and retention (EPR) effect (increased permeability of tumor blood vessels and undeveloped lymphatic system in cancer tissues) The effect of creating an environment that facilitates the accumulation of high-molecular substances due to the construction of a high-molecular-weight cancer tissue that is selectively accumulated in cancer tissue and that is long-term retention in the tumor compared to low-molecular-weight BSH / Because it can be irradiated with thermal neutrons or epithermal neutrons after the 10 B concentration ratio in blood has been achieved, it has been widely studied.
　しかしながら、ナノ粒子型ＤＤＳを用いた場合には完全に均一な腫瘍内分布を達成することが困難であり［Ｓ．Ｓｔａｐｌｅｔｏｎ　ｅｔ　ａｌ．，ＰＬＯＳ　Ｏｎｅ，８：ｅ８１１５７（２０１３）］、繰り返し投与を行う抗がん剤治療では問題とならないが、ホウ素中性子捕捉療法においては、不均一なホウ素化合物の分布は一部のがん細胞の生存へと繋がり、治療効果の低下やがんの再発の原因となる。 However, when nanoparticle type DDS is used, it is difficult to achieve a completely uniform intratumoral distribution [S. Stapleton et al. , PLOS One, 8: e81157 (2013)], which is not a problem with repeated treatment with anticancer drugs, but in boron neutron capture therapy, the uneven distribution of boron compounds results in the survival of some cancer cells. Leading to a decrease in therapeutic effect and cancer recurrence.
　このため、ホウ素中性子捕捉療法のためのＤＤＳには、ホウ素化合物を腫瘍組織内の深部のがん細胞まで送達できる、優れた組織浸透性が必要であると考えられる。 For this reason, it is considered that DDS for boron neutron capture therapy needs to have excellent tissue permeability capable of delivering boron compounds to deep cancer cells in the tumor tissue.
　一方、ホウ素化合物の細胞内移行を重視したＤＤＳ設計としては、カチオン性ポリマー／ホウ素化合物コンジュゲート［Ａ．Ｋ．Ａｚａｂ　ｅｔ　ａｌ．，Ｊ．Ｃｏｎｔｒｏｌ．Ｒｅｌｅａｓｅ，１０６：１４－２５（２００５）；Ｍ．Ｕｍａｎｏ　ｅｔ　ａｌ．，Ａｐｐｌｉｅｄ　Ｒａｄｉａｔｉｏｎ　ａｎｄ　Ｉｓｏｔｏｐｅｓ，６９：１７６５－１７６７（２０１１）］やＣＰＰ／ホウ素化合物コンジュゲート［Ｈ．Ｍｉｃｈｉｕｅ　ｅｔ　ａｌ．，Ｂｉｏｍａｔｅｒｉａｌｓ，３５：３３９６－３４０５（２０１４）］が開発されているが、一般的にこれらのＣＰＰやカチオン性ポリマーは生体分子と非特異的な相互作用を示すために、ＥＰＲ効果によるがん選択的な集積効果が得られにくいと考えられる。 On the other hand, as a DDS design focusing on the intracellular transfer of boron compounds, cationic polymer / boron compound conjugates [A. K. Azab et al. , J .; Control. Release, 106: 14-25 (2005); Umano et al. , Applied Radiation and Isotopes, 69: 1765-1767 (2011)] and CPP / boron compound conjugates [H. Michiue et al. , Biomaterials, 35: 3396-3405 (2014)], but since these CPPs and cationic polymers generally show non-specific interactions with biomolecules, cancer selection based on the EPR effect It is considered that a typical accumulation effect is difficult to obtain.
　また、ＰＥＧを有するカチオン性デンドリマーも開発されているが［Ｂ．Ｑｕａｌｍａｎｎ　ｅｔ　ａｌ．，Ａｎｇｅｗ．　Ｃｈｅｍ．　Ｉｎｔ．　Ｅｄ．，　３５，　９０９－９１１（１９９６）］、多く存在するアミノ基のカチオン性により膜透過性が高く、生体分子に非特異的に吸着し、がん選択的に送達することは困難である。また、腫瘍に均一に分布することは期待できない。さらに、その強い正電荷により、生体内で異物として認識されてしまうことも考えられる。また、ＢＮＣＴで用いる場合、通常の医薬と異なり、多量投与が必要となるが、塩基性アミノ酸のポリマーは、細胞毒性が知られており、多量投与には向いていない。 In addition, a cationic dendrimer having PEG has been developed [B. Qualmann et al. , Angew. Chem. Int. Ed. , 35, 909-911 (1996)], and is highly membrane permeable due to the cationic nature of many amino groups, nonspecifically adsorbing to biomolecules, and difficult to selectively deliver to cancer. In addition, it cannot be expected to be uniformly distributed in the tumor. Furthermore, it may be recognized that the strong positive charge is recognized as a foreign substance in the living body. In addition, when used in BNCT, unlike a normal medicine, a large dose is required. However, a polymer of a basic amino acid is known for cytotoxicity and is not suitable for a large dose.
　本発明の目的は、ＥＰＲ効果による優れたがん集積性、及びナノ粒子化しないことによる腫瘍組織内での高い浸透性を示し、がん細胞内に効率的に取り込まれる新規含ホウ素化合物を提供し、それにより、ホウ素中性子捕捉療法において、効果的な腫瘍疾患の治療を実現することである。 An object of the present invention is to provide a novel boron-containing compound that exhibits excellent cancer accumulation due to the EPR effect, and high penetrability in tumor tissue by not forming nanoparticles, and is efficiently taken into cancer cells. Thus, it is to realize effective treatment of tumor diseases in boron neutron capture therapy.
　本発明者らは、上記課題を解決すべく鋭意検討した結果、ポリエーテル、及びＢＳＨのようなホウ素クラスターを有し、中性アミノ酸及び酸性アミノ酸から選ばれるアミノ酸を構成アミノ酸とするペプチド化合物が、ＥＰＲ効果による優れたがん集積性、及びナノ粒子化しないことによる腫瘍組織内での高い浸透性を示し、さらには、がん細胞内に効率的に取り込まれることを見出し、本発明を完成するに至った。すなわち、本発明は、以下［１］～［３６］の通りである。 As a result of diligent studies to solve the above problems, the present inventors have obtained a peptide compound having a boron cluster such as polyether and BSH, and an amino acid selected from a neutral amino acid and an acidic amino acid as a constituent amino acid. An excellent cancer accumulation property due to the EPR effect and a high permeability in the tumor tissue due to the non-nanoparticulate formation are found, and furthermore, it is found that it can be efficiently taken up into cancer cells, thereby completing the present invention. It came to. That is, the present invention is as follows [1] to [36].
［１］　ホウ素クラスター及びポリエーテルを有し、中性アミノ酸及び酸性アミノ酸から選ばれるアミノ酸を構成アミノ酸とするペプチド化合物又はその塩。 [1] A peptide compound or a salt thereof having a boron cluster and a polyether and having an amino acid selected from a neutral amino acid and an acidic amino acid as a constituent amino acid.
［２］　アミノ酸が構成するペプチドが、直鎖ペプチドである上記［１］に記載のペプチド化合物又はその塩。 [2] The peptide compound or salt thereof according to [1] above, wherein the peptide composed of the amino acid is a linear peptide.
［３］　ホウ素クラスターが側鎖に結合したアミノ酸残基を少なくとも１個有する上記［１］または［２］に記載のペプチド化合物又はその塩。 [3] The peptide compound or salt thereof according to [1] or [2] above, wherein the boron cluster has at least one amino acid residue bonded to the side chain.
［４］　ホウ素クラスターが側鎖に結合したアミノ酸残基が、式（１）： [4] An amino acid residue having a boron cluster bonded to a side chain is represented by the formula (1):
Figure JPOXMLDOC01-appb-C000006
Figure JPOXMLDOC01-appb-C000006
［式中、Ｒ１は、それぞれ独立して、ホウ素クラスターからなる１価の基又はそれがリンカー構造を介して結合する１価の基で置換された、中性アミノ酸及び酸性アミノ酸から選ばれるα－アミノ酸の側鎖を示し、当該α－アミノ酸の側鎖は、それが結合する炭素原子及び該炭素原子に隣接する窒素原子と一緒になって、環を形成していてもよく、環を形成する場合は該窒素原子に結合するＨは存在しない。］
で表される、上記［３］に記載のペプチド化合物又はその塩。 [Wherein, each R 1 independently represents an α selected from a neutral amino acid and an acidic amino acid substituted with a monovalent group consisting of a boron cluster or a monovalent group to which it is bonded via a linker structure. -Indicates the side chain of an amino acid, and the side chain of the α-amino acid may form a ring together with the carbon atom to which it is attached and the nitrogen atom adjacent to the carbon atom. When it does, H couple | bonded with this nitrogen atom does not exist. ]
The peptide compound or salt thereof according to [3] above, represented by:
［５］　Ｒ１が、それぞれ独立して、式（Ｂ１）： [5] R 1 s are each independently represented by formula (B1):
Figure JPOXMLDOC01-appb-C000007
Figure JPOXMLDOC01-appb-C000007
［式中、Ｍ２＋は、カチオンを示し、●は、ＢＨを示し、＊は、リンカー構造との結合部位を示す。］
で表される基がリンカー構造を介して結合する１価の基で置換された、中性アミノ酸及び酸性アミノ酸から選ばれるα－アミノ酸の側鎖である、上記［４］に記載のペプチド化合物又はその塩。 [ Wherein M 2+ represents a cation, ● represents BH, and * represents a binding site with a linker structure. ]
The peptide compound according to the above [4], which is a side chain of an α-amino acid selected from a neutral amino acid and an acidic amino acid, wherein the group represented by formula (1) is substituted with a monovalent group bonded through a linker structure Its salt.
［６］　Ｒ１が、それぞれ独立して、式（Ｒ１ａ）： [6] Each R 1 independently represents the formula (R1a):
Figure JPOXMLDOC01-appb-C000008
Figure JPOXMLDOC01-appb-C000008
［式中、Ｒａは、それぞれ独立して、水素原子、炭素数が１～６のアルキル基で置換されていてもよいカルボキシ基、炭素数が１～６のアルキル基で置換されていてもよいヒドロキシ基、炭素数が１～６のアルキル基で置換されていてもよいメルカプト基、炭素数が６～１４のアリール基、又は炭素数が１～６のアルキル基を示し；Ｑ２は、それぞれ独立して、－Ｏ－、－Ｓ－、－ＮＨ－、－ＳＯ２－、－ＣＯ－、－ＮＨＣＯ－、－ＣＯＮＨ－、－ＯＣＯ－、－ＣＯＯ－、－ＳＣＯ－、－ＣＯＳ－、－ＮＨＣＯＮＨ－、－ＮＨＣＳＮＨ－、－ＯＣＯＮＨ－、－ＮＨＣＯＯ－、－ＳＳ－、－Ｐｈ－、－ＯＰｈＯ－、－ＯＰｈ－、－ＰｈＯ－、－ＳＰｈＳ－、－ＳＰｈ－、－ＰｈＳ－、－ＮＨＰｈＮＨ－、－ＮＨＰｈ－、－ＰｈＮＨ－、－複素環－、－Ｏ－複素環－Ｏ－、－Ｏ－複素環－、－複素環－Ｏ－、－Ｓ－複素環－Ｓ－、－Ｓ－複素環－、－複素環－Ｓ－、－ＮＨ－複素環－ＮＨ－、－ＮＨ－複素環－又は－複素環－ＮＨ－（ここで、Ｐｈは、１，２－フェニレン、１，３－フェニレン又は１，４－フェニレンを示す。）を示し；Ｐ３は、－Ｏ－、－ＮＨ－又は－Ｓ－を示し；Ｍ２＋は、カチオンを示し；●は、ＢＨを示し；ｄは、１又は２を示し；ｆは、それぞれ独立して、１～２０の整数を示し；ｇは、０～２０の整数を示し；＊＊は、α－アミノ酸のα－炭素との結合部位を示す。］
で表される基である、上記［５］に記載のペプチド化合物又はその塩。 [Wherein, each R a is independently a hydrogen atom, a carboxy group optionally substituted with an alkyl group having 1 to 6 carbon atoms, or an alkyl group having 1 to 6 carbon atoms. good hydroxy group, an alkyl mercapto group optionally substituted with a group with a carbon number of 1-6, an aryl group, or an alkyl group having 1 to 6 carbon atoms having a carbon number of 6 ~ 14; Q 2 is Independently, —O—, —S—, —NH—, —SO 2 —, —CO—, —NHCO—, —CONH—, —OCO—, —COO—, —SCO—, —COS—, —NHCONH—, —NHCSNH—, —OCONH—, —NHCOO—, —SS—, —Ph—, —OPhO—, —OPh—, —PhO—, —SPhS—, —SPh—, —PhS—, —NHPhNH -, -NHPh-, -PhNH-, -complex Ring-, -O-heterocycle-O-, -O-heterocycle-, -heterocycle-O-, -S-heterocycle-S-, -S-heterocycle-, -heterocycle-S-,- NH-heterocycle-NH-, -NH-heterocycle- or -heterocycle-NH- (wherein Ph represents 1,2-phenylene, 1,3-phenylene or 1,4-phenylene). P 3 represents —O—, —NH— or —S—; M 2+ represents a cation; ● represents BH; d represents 1 or 2; f represents each independently G represents an integer of 0 to 20; ** represents a binding site with an α-carbon of an α-amino acid. ]
The peptide compound or a salt thereof according to the above [5], which is a group represented by the formula:
［７］　Ｒ１のα－アミノ酸が、グリシン、アラニン、バリン、ロイシン、イソロイシン、セリン、トレオニン、システイン、メチオニン、アスパラギン、グルタミン、フェニルアラニン、チロシン、アスパラギン酸、グルタミン酸、ホモセリン、ノルロイシン、ノルバリン、チロニン、シトルリン、α-アミノ酪酸、ホモシステイン及びペニシラミンからなる群から選ばれる、上記［４］～［６］の何れかに記載のペプチド化合物又はその塩。 [7] The α-amino acid of R 1 is glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, asparagine, glutamine, phenylalanine, tyrosine, aspartic acid, glutamic acid, homoserine, norleucine, norvaline, thyronine, The peptide compound or salt thereof according to any one of [4] to [6] above, selected from the group consisting of citrulline, α-aminobutyric acid, homocysteine and penicillamine.
［８］　Ｒ１のα－アミノ酸が、グルタミン酸及びアスパラギン酸からなる群から選ばれる、上記［４］～［６］の何れかに記載のペプチド化合物又はその塩。 [8] The peptide compound or a salt thereof according to any one of the above [4] to [6], wherein the α-amino acid of R 1 is selected from the group consisting of glutamic acid and aspartic acid.
［９］　ホウ素クラスターが、ペプチドの構成アミノ酸に、直接又はリンカー構造を介して結合する、上記［１］～［３］の何れかに記載のペプチド化合物又はその塩。 [9] The peptide compound or salt thereof according to any one of the above [1] to [3], wherein the boron cluster is bonded to a constituent amino acid of the peptide directly or via a linker structure.
［１０］　リンカー構造が、炭素原子、酸素原子、硫黄原子及び窒素原子から選ばれる１～２００個の主鎖構成原子からなる２価の基である、上記［４］～［９］の何れかに記載のペプチド化合物又はその塩。 [10] Any of the above [4] to [9], wherein the linker structure is a divalent group consisting of 1 to 200 main chain constituent atoms selected from a carbon atom, an oxygen atom, a sulfur atom and a nitrogen atom Or a salt thereof.
［１１］　リンカー構造が、式（Ｌ’）： [11] The linker structure has the formula (L ′):
Figure JPOXMLDOC01-appb-C000009
Figure JPOXMLDOC01-appb-C000009
［式中、Ｒａは、それぞれ独立して、水素原子、炭素数が１～６のアルキル基で置換されていてもよいカルボキシ基、炭素数が１～６のアルキル基で置換されていてもよいヒドロキシ基、炭素数が１～６のアルキル基で置換されていてもよいメルカプト基、炭素数が６～１４のアリール基、又は炭素数が１～６のアルキル基を示し；Ｐ２及びＱ２は、それぞれ独立して、－Ｏ－、－Ｓ－、－ＮＨ－、－ＳＯ２－、－ＣＯ－、－ＮＨＣＯ－、－ＣＯＮＨ－、－ＯＣＯ－、－ＣＯＯ－、－ＳＣＯ－、－ＣＯＳ－、－ＮＨＣＯＮＨ－、－ＮＨＣＳＮＨ－、－ＯＣＯＮＨ－、－ＮＨＣＯＯ－、－ＳＳ－、－Ｐｈ－、－ＯＰｈＯ－、－ＯＰｈ－、－ＰｈＯ－、－ＳＰｈＳ－、－ＳＰｈ－、－ＰｈＳ－、－ＮＨＰｈＮＨ－、－ＮＨＰｈ－、－ＰｈＮＨ－、－複素環－、－Ｏ－複素環－Ｏ－、－Ｏ－複素環－、－複素環－Ｏ－、－Ｓ－複素環－Ｓ－、－Ｓ－複素環－、－複素環－Ｓ－、－ＮＨ－複素環－ＮＨ－、－ＮＨ－複素環－又は－複素環－ＮＨ－（ここで、Ｐｈは、１，２－フェニレン、１，３－フェニレン又は１，４－フェニレンを示す。）を示し；ｆは、それぞれ独立して、１～２０の整数を示し；ｇは、０～２０の整数を示し、＊１はアミノ酸の側鎖との結合部位であり、＊２はホウ素クラスターとの結合部位である。］
で表される２価の基である、上記［４］～［１０］の何れかに記載のペプチド化合物又はその塩。 [Wherein, each R a is independently a hydrogen atom, a carboxy group optionally substituted with an alkyl group having 1 to 6 carbon atoms, or an alkyl group having 1 to 6 carbon atoms. A good hydroxy group, a mercapto group optionally substituted by an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 14 carbon atoms, or an alkyl group having 1 to 6 carbon atoms; P 2 and Q 2 are each independently —O—, —S—, —NH—, —SO 2 —, —CO—, —NHCO—, —CONH—, —OCO—, —COO—, —SCO—, — COS-, -NHCONH-, -NHCSNH-, -OCONH-, -NHCOO-, -SS-, -Ph-, -OPhO-, -OPh-, -PhO-, -SPhS-, -SPh-, -PhS- , -NHPhNH-, -NHPh-, -PhNH , -Heterocycle-, -O-heterocycle-O-, -O-heterocycle-, -heterocycle-O-, -S-heterocycle-S-, -S-heterocycle-, -heterocycle-S -, -NH-heterocycle-NH-, -NH-heterocycle- or -heterocycle-NH- (wherein Ph represents 1,2-phenylene, 1,3-phenylene or 1,4-phenylene F is independently an integer of 1 to 20; g is an integer of 0 to 20, * 1 is a binding site with an amino acid side chain, and * 2 is boron. This is the binding site with the cluster. ]
The peptide compound or salt thereof according to any one of [4] to [10] above, which is a divalent group represented by the formula:
［１２］　ペプチド化合物のＣ末端のカルボキシ基及び／又はペプチド化合物のＮ末端のアミノ基がポリエーテルからなる１価の基で置換されている、上記［１］～［１１］の何れかに記載のペプチド化合物又はその塩。 [12] Any one of [1] to [11] above, wherein the C-terminal carboxy group of the peptide compound and / or the N-terminal amino group of the peptide compound is substituted with a monovalent group comprising a polyether. Or a salt thereof.
［１３］　ペプチド化合物のＣ末端のカルボキシ基及び／又はペプチド化合物のＮ末端のアミノ基が、式（Ｐ１）： [13] The carboxy group at the C-terminal of the peptide compound and / or the amino group at the N-terminal of the peptide compound are represented by the formula (P1):
Figure JPOXMLDOC01-appb-C000010
Figure JPOXMLDOC01-appb-C000010
［式中、Ｚは、置換対象がカルボキシ基の場合－Ｏ－、－ＮＨ－又は－Ｓ－を、置換対象がアミノ基の場合－ＣＯ－、－ＣＯＮＨ－又は－ＣＯＯ－を示し、Ｒｄ及びＲｅは、それぞれ独立して水素原子又は炭素数が１～６のアルキル基を示し、ｓは、それぞれ独立して１～１０の整数であり、ｔは、それぞれ独立して２～１０の整数であり、ｕは、それぞれ独立して１以上の整数であり、＊＊＊は、結合部位を示す。］
で表される基で置換されている、上記［１］～［１２］の何れかに記載のペプチド化合物又はその塩。 [In the formula, Z represents —O—, —NH— or —S— when the substitution target is a carboxy group, and —CO—, —CONH— or —COO— when the substitution target is an amino group, R d And R e each independently represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, s is each independently an integer of 1 to 10, and t is independently 2 to 10 Each is an integer, u is each independently an integer of 1 or more, and *** represents a binding site. ]
The peptide compound or a salt thereof according to any one of the above [1] to [12], which is substituted with a group represented by:
［１４］　ホウ素クラスターが、３～２０個のホウ素原子を有するホウ素クラスターである、上記［１］～［１３］の何れかに記載のペプチド化合物又はその塩。 [14] The peptide compound or salt thereof according to any one of [1] to [13] above, wherein the boron cluster is a boron cluster having 3 to 20 boron atoms.
［１５］　ホウ素クラスターが、水素化ホウ素化合物、及びその１個以上のホウ素原子がそれぞれ独立して炭素原子又は窒素原子で置換された化合物、並びにそれらの金属錯体からなる群から選ばれる、上記［１］～［１４］の何れかに記載のペプチド化合物又はその塩。 [15] The above, wherein the boron cluster is selected from the group consisting of a borohydride compound, a compound in which one or more boron atoms are each independently substituted with a carbon atom or a nitrogen atom, and a metal complex thereof. [1] The peptide compound or salt thereof according to any one of [14] to [14].
［１６］　ホウ素クラスターの１０Ｂの同位体比が、５０%以上である、上記［１］～［１５］の何れかに記載のペプチド化合物又はその塩。 [16] The peptide compound or salt thereof according to any one of [1] to [15] above, wherein the 10 B isotope ratio of the boron cluster is 50% or more.
［１７］　ペプチドがポリアミノ酸である上記［１］～［１６］の何れかに記載のペプチド化合物又はその塩。 [17] The peptide compound or salt thereof according to any one of [1] to [16] above, wherein the peptide is a polyamino acid.
［１８］　ペプチドのアミノ酸残基数に対するホウ素クラスターの結合数の比が０．１以上である、上記［１］～［１７］の何れかに記載のペプチド化合物又はその塩。 [18] The peptide compound or salt thereof according to any one of [1] to [17] above, wherein the ratio of the number of bonds of boron clusters to the number of amino acid residues of the peptide is 0.1 or more.
［１９］　ペプチドのアミノ酸残基数が２００以下である、上記［１］～［１８］の何れかに記載のペプチド化合物又はその塩。 [19] The peptide compound or salt thereof according to any one of [1] to [18] above, wherein the peptide has 200 or less amino acid residues.
［２０］　ポリエーテルが、ポリアルキレングリコール単位を含む直鎖又は分枝鎖の水溶性重合体である、上記［１］～［１９］の何れかに記載のペプチド化合物又はその塩。 [20] The peptide compound or salt thereof according to any one of [1] to [19] above, wherein the polyether is a linear or branched water-soluble polymer containing a polyalkylene glycol unit.
［２１］　ポリアルキレングリコール単位が、ポリエチレングリコール単位である、上記［２０］に記載のペプチド化合物又はその塩。 [21] The peptide compound or salt thereof according to [20] above, wherein the polyalkylene glycol unit is a polyethylene glycol unit.
［２２］　ポリエーテルの数平均分子量が、５０，０００未満である、上記［１］～［２１］の何れかに記載のペプチド化合物又はその塩。 [22] The peptide compound or salt thereof according to any one of [1] to [21] above, wherein the polyether has a number average molecular weight of less than 50,000.
［２３］　ペプチドの構成アミノ酸が、側鎖にカルボキシ基、ヒドロキシ基及びメルカプト基から選ばれる活性基を有するアミノ酸である、上記［１］～［２２］の何れかに記載のペプチド化合物又はその塩。 [23] The peptide compound or a salt thereof according to any one of [1] to [22] above, wherein the constituent amino acid of the peptide is an amino acid having an active group selected from a carboxy group, a hydroxy group and a mercapto group in the side chain. .
［２４］　ペプチドの構成アミノ酸が、グルタミン酸及びアスパラギン酸から選ばれる、上記［１］～［２３］の何れかに記載のペプチド化合物又はその塩。 [24] The peptide compound or salt thereof according to any one of [1] to [23] above, wherein the constituent amino acids of the peptide are selected from glutamic acid and aspartic acid.
［２５］　分子量が、１００，０００未満である、上記［１］～［２４］の何れかに記載のペプチド化合物又はその塩。 [25] The peptide compound or salt thereof according to any one of [1] to [24] above, wherein the molecular weight is less than 100,000.
［２６］　式（Ｉａ）又は式（Ｉｂ）： [26] Formula (Ia) or Formula (Ib):
Figure JPOXMLDOC01-appb-C000011
Figure JPOXMLDOC01-appb-C000011
［式中、
　Ｒ１は、それぞれ独立して、ホウ素クラスターからなる１価の基又はそれがリンカー構造を介して結合する１価の基で置換された、中性アミノ酸及び酸性アミノ酸から選ばれるα－アミノ酸の側鎖を示し、
　Ｒ２は、それぞれ独立して、置換基を有していてもよい、中性アミノ酸及び酸性アミノ酸から選ばれるα－アミノ酸の側鎖を示し、
　Ｒ１及びＲ２のα－アミノ酸の側鎖は、それぞれ、それが結合する炭素原子及び該炭素原子に隣接する窒素原子と一緒になって、環を形成していてもよく、環を形成する場合は該窒素原子に結合するＨは存在せず、
　ｂが１以上の場合、Ｒ１を有するα－アミノ酸残基とＲ２を有するα－アミノ酸残基とは、任意に共重合しており、
　Ｘ１、及びｃが１の場合のＹ２は、それぞれ独立して、
（１）水素原子、
（２）炭素数が１～６のアルキル基、
（３）炭素数が２～７のアルキルカルボニル基、又は
（４）ポリエーテルからなる１価の基を示し、
　Ｘ２、及びｃが１の場合のＹ１は、それぞれ独立して、
（１）炭素数が１～６のアルキル基で置換されていてもよいヒドロキシ基、
（２）炭素数が１～６のアルキル基で置換されていてもよいアミノ基、又は
（３）ポリエーテルからなる１価の基を示し、
　ｃが２以上の場合のＹ１及びＹ２は、それぞれ、ポリエーテルからなるｃ価の基を示し、
　式（Ｉａ）においては、Ｒ２のα－アミノ酸の側鎖の置換基、並びにＸ１及びＹ１のうち少なくともいずれか１つが、式（Ｉｂ）においては、Ｒ２のα－アミノ酸の側鎖の置換基、並びにＸ２及びＹ２のうち少なくともいずれか１つが、ポリエーテルからなる１価の基又はポリエーテルからなるｃ価の基であり、
　ａ及びｃは、それぞれ独立して、１以上の整数を示し、
　ｂは、０以上の整数を示し、
　ａとｂの合計は、２以上である。］
で表される、上記［１］～［２５］の何れかに記載のペプチド化合物又はその塩。 [Where:
R 1 is each independently an α-amino acid selected from a neutral amino acid and an acidic amino acid substituted with a monovalent group consisting of a boron cluster or a monovalent group to which it is bonded via a linker structure Showing the chain
R 2 each independently represents a side chain of an α-amino acid selected from a neutral amino acid and an acidic amino acid, which may have a substituent,
The side chain of the α-amino acid of R 1 and R 2 may be combined with the carbon atom to which it is attached and the nitrogen atom adjacent to the carbon atom to form a ring. In this case, there is no H bonded to the nitrogen atom,
When b is 1 or more, the α-amino acid residue having R 1 and the α-amino acid residue having R 2 are optionally copolymerized,
X 1 and Y 2 when c is 1 are each independently
(1) a hydrogen atom,
(2) an alkyl group having 1 to 6 carbon atoms,
(3) an alkylcarbonyl group having 2 to 7 carbon atoms, or (4) a monovalent group comprising a polyether,
X 2 and Y 1 when c is 1 are each independently
(1) a hydroxy group which may be substituted with an alkyl group having 1 to 6 carbon atoms,
(2) an amino group which may be substituted with an alkyl group having 1 to 6 carbon atoms, or (3) a monovalent group comprising a polyether,
Y 1 and Y 2 in the case where c is 2 or more each represent a c-valent group composed of a polyether;
In formula (Ia), at least one of the substituents on the side chain of the α-amino acid of R 2 and X 1 and Y 1 is the side chain of the α-amino acid of R 2 in formula (Ib) And at least one of X 2 and Y 2 is a monovalent group made of a polyether or a c-valent group made of a polyether,
a and c each independently represent an integer of 1 or more,
b represents an integer of 0 or more,
The sum of a and b is 2 or more. ]
The peptide compound or a salt thereof according to any one of the above [1] to [25] represented by the formula:
［２７］　ｃが１である、上記［２６］に記載のペプチド化合物又はその塩。 [27] The peptide compound or salt thereof according to [26] above, wherein c is 1.
［２８］　Ｙ１及びＹ２が、ポリエーテルからなる１価の基である、上記［２７］に記載のペプチド化合物又はその塩。 [28] The peptide compound or a salt thereof according to the above [27], wherein Y 1 and Y 2 are a monovalent group consisting of a polyether.
［２９］　Ｙ１及びＹ２が、式（Ｐ１）： [29] Y 1 and Y 2 are represented by the formula (P1):
Figure JPOXMLDOC01-appb-C000012
Figure JPOXMLDOC01-appb-C000012
［式中、Ｚは、式（Ｉａ）においては－Ｏ－、－ＮＨ－又は－Ｓ－を、式（Ｉｂ）においては－ＣＯ－、－ＣＯＮＨ－又は－ＣＯＯ－を示し、Ｒｄ及びＲｅは、それぞれ独立して水素原子又は炭素数が１～６のアルキル基を示し、ｓは、それぞれ独立して１～１０の整数であり、ｔは、それぞれ独立して２～１０の整数であり、ｕは、それぞれ独立して１以上の整数であり、＊＊＊は、結合部位を示す。］
で表される基で置換されている、上記［２７］に記載のペプチド化合物又はその塩。 [Wherein Z represents —O—, —NH— or —S— in formula (Ia), —CO—, —CONH— or —COO— in formula (Ib), R d and R e is independently a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, s is independently an integer of 1 to 10, and t is independently an integer of 2 to 10; Each u is independently an integer of 1 or more, and *** represents a binding site. ]
The peptide compound or a salt thereof according to the above [27], which is substituted with a group represented by the formula:
［３０］　Ｘ１が、水素原子であり、Ｘ２が、炭素数が１～６のアルキル基で置換されていてもよいヒドロキシ基、又は炭素数が１～６のアルキル基で置換されていてもよいアミノ基である、上記［２６］～［２９］の何れかに記載のペプチド化合物又はその塩。 [30] X 1 is a hydrogen atom, and X 2 is a hydroxy group which may be substituted with an alkyl group having 1 to 6 carbon atoms, or an alkyl group having 1 to 6 carbon atoms. The peptide compound or a salt thereof according to any one of the above [26] to [29], which is a good amino group.
［３１］　ａとｂの合計が、２００以下の整数である、上記［２６］～［３０］の何れかに記載のペプチド化合物又はその塩。 [31] The peptide compound or salt thereof according to any one of [26] to [30] above, wherein the sum of a and b is an integer of 200 or less.
［３２］　ａとｂの合計に対するａの比が、０．１以上である、上記［２６］～［３１］の何れかに記載のペプチド化合物又はその塩。 [32] The peptide compound or salt thereof according to any one of [26] to [31], wherein the ratio of a to the sum of a and b is 0.1 or more.
［３３］　腫瘍疾患のホウ素中性子捕捉療法に用いるための上記［１］～［３２］の何れかに記載のペプチド化合物又はその塩。 [33] The peptide compound or salt thereof according to any one of [1] to [32] for use in boron neutron capture therapy for tumor diseases.
［３４］　上記［１］～［３２］の何れかに記載のペプチド化合物又はその塩、及び薬学的に許容される担体を含む医薬組成物。 [34] A pharmaceutical composition comprising the peptide compound or a salt thereof according to any one of [1] to [32] above, and a pharmaceutically acceptable carrier.
［３５］　腫瘍疾患のホウ素中性子捕捉療法に用いるための医薬を製造するための上記［１］～［３２］の何れかに記載のペプチド化合物又はその塩の使用。 [35] Use of the peptide compound or salt thereof according to any one of [1] to [32] above for producing a medicament for use in boron neutron capture therapy for tumor diseases.
［３６］　上記［１］～［３２］の何れかに記載のペプチド化合物又はその塩の必要量を哺乳動物に投与することを特徴とするホウ素中性子捕捉療法による腫瘍疾患の治療方法。 [36] A method for treating a tumor disease by boron neutron capture therapy, comprising administering a necessary amount of the peptide compound or salt thereof according to any one of [1] to [32] to a mammal.
　本発明の化合物は、ＥＰＲ効果による優れたがん集積性、及びナノ粒子化しないことによる腫瘍組織内での高い浸透性を示し、がん細胞内に効率的に取り込まれるため、これを用いるホウ素中性子捕捉療法ではより効果的に腫瘍疾患を治療できる。 The compound of the present invention exhibits excellent cancer accumulation due to the EPR effect, and high penetrability in tumor tissues due to non-nanoparticulation, and is efficiently incorporated into cancer cells. Neutron capture therapy can treat tumor diseases more effectively.
ＨＵＶＥＣに対する細胞毒性試験における実施例１～３及び比較例１及び２の化合物並びに１０Ｂ－ＢＳＨの濃度別の細胞生存率に関するグラフを示す。The graph regarding the cell viability according to the density | concentration of the compound of Examples 1-3 and the comparative examples 1 and 2 and 10 B-BSH in the cytotoxicity test with respect to HUVEC is shown. Ｃ２６がん細胞に対する細胞毒性試験における実施例１～３及び比較例１及び２の化合物並びに１０Ｂ－ＢＳＨの濃度別の細胞生存率に関するグラフを示す。2 is a graph showing the cell viability according to the concentrations of the compounds of Examples 1 to 3 and Comparative Examples 1 and 2 and 10 B-BSH in the cytotoxicity test for C26 cancer cells. 実施例１～３及び比較例１及び２の化合物並びに１０Ｂ－ＢＳＨのＣ２６がん細胞内取込量に関するグラフを示す。グラフの縦軸は、１０Ｂ－ＢＳＨをＣ２６がん細胞に添加後１時間の細胞内取込量を１とした時の、各化合物の相対的な細胞内取込量を示す。The graph regarding the uptake | capture amount in the C26 cancer cell of the compound of Examples 1-3 and the comparative examples 1 and 2, and 10 B-BSH is shown. The vertical axis of the graph shows the relative cellular uptake of each compound when the amount of cellular uptake for 1 hour after addition of 10 B-BSH to C26 cancer cells is 1. 実施例１の化合物をＡｌｅｘａ４８８（登録商標）で蛍光標識した化合物を添加したＣ２６がん細胞の共焦点レーザー顕微鏡（ＣＬＳＭ）による画像を示す。The image by the confocal laser scanning microscope (CLSM) of the C26 cancer cell which added the compound which fluorescently labeled the compound of Example 1 with Alexa488 (trademark) is shown. 実施例７の化合物、及び合成例２１の化合物の対カチオンが２セシウムカチオンである化合物のＣＴ２６がん細胞内取込量に関するグラフを示す。The graph regarding the amount of CT26 cancer cell uptake | capture of the compound whose counter cation of the compound of Example 7 and the compound of the synthesis example 21 is a 2 cesium cation is shown. 実施例１～３若しくは比較例１若しくは２の化合物又は１０Ｂ－ＢＳＨを、Ｃ２６がん細胞を皮下移植したマウスに投与した後の血漿中濃度推移に関するグラフを示す。グラフの縦軸は、１　ｍｌの血漿当り、投与量（ＩＤ：ｉｎｊｅｃｔｅｄ　ｄｏｓｅ）の何％が送達されたかを示す。2 is a graph relating to plasma concentration transitions after administration of the compounds of Examples 1 to 3 or Comparative Example 1 or 2 or 10 B-BSH to mice transplanted subcutaneously with C26 cancer cells. The vertical axis of the graph shows what percentage of the dose (ID) delivered per ml of plasma. 実施例１～３若しくは比較例１若しくは２の化合物又は１０Ｂ－ＢＳＨを、Ｃ２６がん細胞を皮下移植したマウスに投与した後の腫瘍細胞中の集積量推移に関するグラフを示す。グラフの縦軸は、１　ｇの腫瘍当り、投与量（ＩＤ：ｉｎｊｅｃｔｅｄ　ｄｏｓｅ）の何％が送達されたかを示す。FIG. 3 shows a graph regarding the change in the amount of accumulation in tumor cells after administering the compound of Examples 1 to 3 or Comparative Example 1 or 2 or 10 B-BSH to mice transplanted subcutaneously with C26 cancer cells. The vertical axis of the graph shows what percentage of the dose (ID) delivered per gram of tumor. 実施例７の化合物又は天然型Ｂ－ＢＳＨを、ＣＴ２６がん細胞を皮下移植したマウスに投与した後の腫瘍細胞中の集積量推移に関するグラフを示す。グラフの縦軸は、１　ｇの腫瘍当り、投与量（ＩＤ：ｉｎｊｅｃｔｅｄ　ｄｏｓｅ）の何％が送達されたかを示す。2 is a graph relating to the change in the amount of accumulation in tumor cells after the compound of Example 7 or natural B-BSH was administered to mice subcutaneously transplanted with CT26 cancer cells. The vertical axis of the graph shows what percentage of the dose (ID: injected dose) was delivered per 1 g tumor. 実施例１の化合物又は１０Ｂ－ＢＳＨを、Ｃ２６がん細胞を皮下移植したマウスに投与及び熱中性子線を１時間照射後又は照射なしの場合の相対腫瘍体積推移に関するグラフを示す。The graph regarding the relative tumor volume transition when the compound of Example 1 or 10 B-BSH is administered to a mouse transplanted with C26 cancer cells subcutaneously and after irradiation with thermal neutrons for 1 hour or without irradiation is shown. 実施例１の化合物又は１０Ｂ－ＢＳＨを、Ｃ２６がん細胞を皮下移植したマウスに投与及び熱中性子線を１時間照射後又は照射なしの場合のマウスの体重推移に関するグラフを示す。The graph regarding the body weight transition of the mouse | mouth when the compound of Example 1 or 10 B-BSH is administered to the mouse | mouth transplanted subcutaneously with the C26 cancer cell, and a thermal neutron ray is irradiated for 1 hour, or is not irradiated is shown. 実施例１の化合物と、リポソーム製剤について、担がんマウスを用いた腫瘍浸透性の比較に関する、共焦点レーザー顕微鏡（ＣＬＳＭ）による画像を示す。左上図は、Ｃ２６腫瘍組織のヘマトキシリン－エオシン染色を示す。左下図は、実施例１の化合物をＡｌｅｘａ６４７（登録商標）で蛍光標識した化合物とＤｏｘｉｌ（登録商標）のＣ２６腫瘍組織分布を示す。中央図は、ＢｘＰＣ３腫瘍組織のヘマトキシリン－エオシン染色を示す。右図は、実施例１の化合物をＡｌｅｘａ６４７（登録商標）で蛍光標識した化合物とＤｏｘｉｌ（登録商標）のＢｘＰＣ３腫瘍組織分布を示す。実施例１の化合物をＡｌｅｘａ６４７（登録商標）で蛍光標識した化合物は緑色、Ｄｏｘｉｌ（登録商標）は赤色、両化合物が共在する部分は黄色として示される。The image by a confocal laser microscope (CLSM) regarding the comparison of the tumor permeability | transmittance using a cancer-bearing mouse | mouth about the compound of Example 1 and a liposome formulation is shown. The upper left figure shows hematoxylin-eosin staining of C26 tumor tissue. The lower left figure shows the C26 tumor tissue distribution of the compound of Example 1 fluorescently labeled with Alexa647 (registered trademark) and Doxil (registered trademark). The middle panel shows hematoxylin-eosin staining of BxPC3 tumor tissue. The right figure shows the BxPC3 tumor tissue distribution of the compound of Example 1 fluorescently labeled with Alexa647 (registered trademark) and Doxil (registered trademark). A compound obtained by fluorescently labeling the compound of Example 1 with Alexa 647 (registered trademark) is shown in green, Doxil (registered trademark) in red, and a portion where both compounds coexist is shown in yellow.
　本発明は、ホウ素クラスター及びポリエーテルを有し、中性アミノ酸及び酸性アミノ酸から選ばれるアミノ酸を構成アミノ酸とするペプチド化合物（以下、「本発明のペプチド化合物」という）又はその塩を提供する。 The present invention provides a peptide compound having a boron cluster and a polyether and having an amino acid selected from a neutral amino acid and an acidic amino acid as a constituent amino acid (hereinafter referred to as “the peptide compound of the present invention”) or a salt thereof.
　本明細書において「ペプチド」とは、２個以上のアミノ酸がペプチド結合によって結合したものを意味する。ペプチドのアミノ酸残基数は、好ましくは、３以上であり、より好ましくは、５以上であり、さらに好ましくは、１０以上である。ペプチドのアミノ酸残基数は、好ましくは、２００以下であり、より好ましくは、１５０以下であり、さらに好ましくは、１００以下である。 In the present specification, “peptide” means a peptide in which two or more amino acids are bound by peptide bonds. The number of amino acid residues of the peptide is preferably 3 or more, more preferably 5 or more, and still more preferably 10 or more. The number of amino acid residues of the peptide is preferably 200 or less, more preferably 150 or less, and still more preferably 100 or less.
　本明細書において「ペプチド化合物」とは、任意の置換基を有していてもよいペプチドを意味する。ペプチド化合物のペプチドは、多くのホウ素クラスターを結合させるという観点から、ペプチドデンドリマーや分岐ペプチドではなく、直鎖ペプチドであることが好ましい。 In the present specification, the “peptide compound” means a peptide which may have an arbitrary substituent. The peptide of the peptide compound is preferably a linear peptide, not a peptide dendrimer or a branched peptide, from the viewpoint of binding many boron clusters.
　本明細書において「アミノ酸」とは、天然型又は非天然型の公知のアミノ酸を意味する。 In the present specification, “amino acid” means a known amino acid of natural type or non-natural type.
　本明細書において「中性アミノ酸」とは、天然型又は非天然型の公知の中性アミノ酸を意味する。例えば、グリシン、アラニン、バリン、ロイシン、イソロイシン、セリン、トレオニン、システイン、メチオニン、アスパラギン、グルタミン、フェニルアラニン、チロシン、ホモセリン、ノルロイシン、ノルバリン、チロニン、シトルリン、α-アミノ酪酸、ホモシステイン、ペニシラミン、プロリン、ヒドロキシプロリン、β－アラニン、γ-アミノ酪酸等である。 In the present specification, “neutral amino acid” means a known neutral amino acid of natural type or non-natural type. For example, glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, asparagine, glutamine, phenylalanine, tyrosine, homoserine, norleucine, norvaline, thyronine, citrulline, α-aminobutyric acid, homocysteine, penicillamine, proline, Hydroxyproline, β-alanine, γ-aminobutyric acid and the like.
　本明細書において「酸性アミノ酸」とは、天然型又は非天然型の公知の酸性アミノ酸を意味する。例えば、アスパラギン酸、グルタミン酸等である。 As used herein, “acidic amino acid” means a known acidic amino acid of natural type or non-natural type. For example, aspartic acid and glutamic acid.
　ペプチドの構成アミノ酸は、好ましくは、セリン、トレオニン、システイン、チロシン、アスパラギン酸、グルタミン酸、ホモセリン、ホモシステイン、ペニシラミン、ヒドロキシプロリン等の側鎖にカルボキシ基、ヒドロキシ基、メルカプト基等の活性基を有するアミノ酸であり；なかでも、グルタミン酸及びアスパラギン酸から選ばれることがより好ましく；グルタミン酸であることが特に好ましい。 The constituent amino acids of the peptide preferably have an active group such as carboxy group, hydroxy group, mercapto group in the side chain of serine, threonine, cysteine, tyrosine, aspartic acid, glutamic acid, homoserine, homocysteine, penicillamine, hydroxyproline, etc. Among them, it is more preferable that it is selected from glutamic acid and aspartic acid, and it is particularly preferable that it is glutamic acid.
　これらのアミノ酸は、Ｄ型、Ｌ型又はそれらの混合物であってもよい。 These amino acids may be D-type, L-type or a mixture thereof.
　ある実施形態では、ペプチドがポリアミノ酸である。即ち、ペプチドの構成アミノ酸がすべて同一のアミノ酸である。 In certain embodiments, the peptide is a polyamino acid. That is, all the constituent amino acids of the peptide are the same amino acid.
　ある実施形態では、ペプチド化合物は、ホウ素クラスターが側鎖に結合したアミノ酸残基を少なくとも１個有する。ペプチドのアミノ酸残基数に対する、ホウ素クラスターが側鎖に結合したアミノ酸残基数の比は、好ましくは、０．１以上であり、より好ましくは、０．２以上であり、さらに好ましくは、０．３以上である。 In one embodiment, the peptide compound has at least one amino acid residue having a boron cluster bonded to a side chain. The ratio of the number of amino acid residues in which the boron cluster is bonded to the side chain to the number of amino acid residues of the peptide is preferably 0.1 or more, more preferably 0.2 or more, and still more preferably 0. .3 or more.
　本明細書において「アミノ酸残基」とは、ペプチドの構成アミノ酸の１単位であって、アミノ酸のＣ末端のＯＨ及び／又はＮ末端のＨを除いた基を意味する。本明細書においてアミノ酸の「側鎖」とは、ペプチドを構成する場合に、その主鎖を構成しない分岐鎖を意味する。 In the present specification, the “amino acid residue” is a unit of a constituent amino acid of a peptide, and means a group excluding OH at the C-terminal of the amino acid and / or H at the N-terminal. In the present specification, the “side chain” of an amino acid means a branched chain that does not constitute the main chain when constituting a peptide.
　本明細書において「ホウ素クラスター」とは、主にホウ素原子が複数集合して一つに結合して得られる構造を有するイオン性又は非イオン性の公知物質を意味し、３～２０個のホウ素原子を有するホウ素クラスターであることが好ましく、８～２０個のホウ素原子を有するホウ素クラスターであることがより好ましく、１０～１２個のホウ素原子を有するホウ素クラスターであることが特に好ましい。 In the present specification, the “boron cluster” means an ionic or nonionic known substance having a structure obtained mainly by combining a plurality of boron atoms and combining them into 3 to 20 boron atoms. A boron cluster having atoms is preferable, a boron cluster having 8 to 20 boron atoms is more preferable, and a boron cluster having 10 to 12 boron atoms is particularly preferable.
　ホウ素クラスターは、それを構成する骨格原子として、ホウ素原子以外に、炭素原子、窒素原子、酸素原子、硫黄原子等を含んでいてもよい。ホウ素クラスターにおいて、炭素原子、窒素原子、酸素原子、硫黄原子等のその他の原子の数は、好ましくは、０～５であり、より好ましくは０～２である。 The boron cluster may contain a carbon atom, a nitrogen atom, an oxygen atom, a sulfur atom, and the like as a skeleton atom constituting the boron cluster. In the boron cluster, the number of other atoms such as carbon atom, nitrogen atom, oxygen atom and sulfur atom is preferably 0 to 5, and more preferably 0 to 2.
　また、ホウ素クラスターは、ニッケルやコバルト等の遷移金属との金属錯体であってもよい。 Further, the boron cluster may be a metal complex with a transition metal such as nickel or cobalt.
　ホウ素クラスターは、イオン性であることが好ましく、また、水溶性であることが好ましい。 The boron cluster is preferably ionic and preferably water-soluble.
　ホウ素クラスターとしては、例えば、［Ｂ３Ｈ８］－Ｍ＋、［Ｂ６Ｈ６］２－Ｍ２＋、［Ｂ７Ｈ７］２－Ｍ２＋、［Ｂ８Ｈ８］２－Ｍ２＋、［Ｂ９Ｈ９］２－Ｍ２＋、［Ｂ１０Ｈ１０］２－Ｍ２＋、［Ｂ１０Ｈ１３］－Ｍ＋、［Ｂ１１Ｈ１１］２－Ｍ２＋、［Ｂ１１Ｈ１４］－Ｍ＋、［Ｂ１２Ｈ１２］２－Ｍ２＋、［Ｂ２０Ｈ１８］４－Ｍ４＋（式中、Ｍ＋、Ｍ２＋及びＭ４＋はそれぞれカチオンを示す。）のような水素化ホウ素化合物、及びそれらの１個以上（好ましくは１～５個、より好ましくは１又は２個）のホウ素原子がそれぞれ独立して炭素原子又は窒素原子で置換された化合物（例えば、［Ｃ２Ｂ９Ｈ１１］－Ｍ＋、［ＣＢ１１Ｈ１２］－Ｍ＋、Ｃ２Ｂ１０Ｈ１２、ＮＢ８Ｈ９）、並びにそれらの金属錯体（例えばニッケル錯体、コバルト錯体）（例えば、［（Ｃ２Ｂ９Ｈ１１）２Ｃｏ］－Ｍ＋、［（Ｃ２Ｂ９Ｈ１１）２Ｎｉ］－Ｍ＋）が挙げられる。 Examples of the boron cluster include [B 3 H 8 ] − M + , [B 6 H 6 ] 2− M 2+ , [B 7 H 7 ] 2 − M 2+ , [B 8 H 8 ] 2 − M 2+ , [B 9 H 9 ] 2− M 2+ , [B 10 H 10 ] 2− M 2+ , [B 10 H 13 ] − M + , [B 11 H 11 ] 2− M 2+ , [B 11 H 14 ] − Boron hydride compounds such as M + , [B 12 H 12 ] 2− M 2+ , [B 20 H 18 ] 4− M 4+ , wherein M + , M 2+ and M 4+ each represent a cation. And compounds in which one or more (preferably 1 to 5, more preferably 1 or 2) boron atoms are independently substituted with carbon atoms or nitrogen atoms (for example, [C 2 B 9 H 11] - M +, [CB 11 H 12] - M +, 2 B 10 H 12, NB 8 H 9), and their metal complexes (e.g. nickel complex, cobalt complex) (e.g., [(C 2 B 9 H 11) 2 Co] - M +, [(C 2 B 9 H 11 ) 2 Ni] − M + ).
　具体例としては、下記式で表されるホウ素クラスターが挙げられる。 Specific examples include boron clusters represented by the following formula.
Figure JPOXMLDOC01-appb-C000013
Figure JPOXMLDOC01-appb-C000013
［式中、●はＢＨを示し、その他の記号は上記定義の通りである。］ [In the formula, ● represents BH, and the other symbols are as defined above. ]
　中でも好ましくは、下記式で表されるホウ素クラスターである。 Among them, a boron cluster represented by the following formula is preferable.
Figure JPOXMLDOC01-appb-C000014
Figure JPOXMLDOC01-appb-C000014
［式中、各記号は上記定義の通りである。］ [Wherein the symbols are as defined above. ]
　Ｍ＋、Ｍ２＋又はＭ４＋で表されるカチオンは、医薬上許容されるカチオンであればよく、特に限定されるものではない。Ｍ＋、Ｍ２＋及びＭ４＋は、単一成分であってもよいし、２種以上の成分の組み合わせであってもよく、それぞれのカチオンは１価であっても多価であってもよい。好ましくは、水素イオン、アルカリ金属イオン（例えば、ナトリウムイオン、カリウムイオン、セシウムイオン等）、アルカリ土類金属イオン（例えば、マグネシウムイオン、カルシウムイオン等）、アミン系イオン（例えば、テトラメチルアンモニウムイオン、テトラブチルアンモニウムイオン等）、ポリアミン（例えば、１，４－ジアミノブタン、スペルミン、スペルミジン）の１価又は多価のアンモニウムイオン等から選ばれるカチオンであり、より好ましくは、アルカリ金属イオンであり、特に好ましくは、ナトリウムイオンである。 The cation represented by M + , M 2+ or M 4+ is not particularly limited as long as it is a pharmaceutically acceptable cation. M + , M 2+ and M 4+ may be a single component or a combination of two or more components, and each cation may be monovalent or polyvalent. . Preferably, hydrogen ions, alkali metal ions (for example, sodium ions, potassium ions, cesium ions, etc.), alkaline earth metal ions (for example, magnesium ions, calcium ions, etc.), amine ions (for example, tetramethylammonium ions, A cation selected from monovalent or polyvalent ammonium ions of polyamines (for example, 1,4-diaminobutane, spermine, spermidine), more preferably alkali metal ions, Sodium ion is preferable.
　ホウ素クラスターの１０Ｂの同位体比は、好ましくは、１９％以上であり、より好ましくは、５０%以上であり、さらに好ましくは、８０%以上である。 The 10 B isotope ratio of the boron cluster is preferably 19% or more, more preferably 50% or more, and still more preferably 80% or more.
　本発明のペプチド化合物において、ホウ素クラスターは、例えば、ペプチドの構成アミノ酸に、直接又はリンカー構造を介して結合する。ホウ素クラスターは、ペプチドの構成アミノ酸の側鎖に１個以上結合していることが好ましく、１～３個がより好ましく、１個が特に好ましい。 In the peptide compound of the present invention, the boron cluster is bonded to, for example, a constituent amino acid of the peptide directly or via a linker structure. The boron cluster is preferably bonded to one or more side chains of the constituent amino acids of the peptide, more preferably 1 to 3, and particularly preferably 1.
　ある好ましい実施形態では、ホウ素クラスターは、アミノ酸の側鎖に、１価の基（ホウ素クラスターの１個の任意のＨを除いてなる基を意味し、以下「ホウ素クラスターからなる１価の基」という）として、直接、或いはリンカー構造を介して、共有結合していることがより好ましい。 In a preferred embodiment, the boron cluster is a monovalent group (meaning a group formed by removing any one H of a boron cluster from the side chain of an amino acid, and hereinafter referred to as “monovalent group consisting of a boron cluster”). More preferably, it is covalently bonded directly or via a linker structure.
　ある実施形態では、ホウ素クラスター（又はリンカー構造）は、例えば、アミノ酸の側鎖の炭素原子、窒素原子又は硫黄原子上の任意のＨ又はＯＨと置換して結合しており、アミノ酸の活性基のＨ及びＯＨ（例えば、カルボキシ基のＯＨ並びにヒドロキシ基及びメルカプト基のＨ）のから選ばれる基と置換して結合していることがより好ましく、カルボキシ基のＯＨと置換して結合していることがさらに好ましい。 In some embodiments, the boron cluster (or linker structure) is attached, for example, by substitution with any H or OH on the carbon, nitrogen or sulfur atom of the side chain of the amino acid, and the active group of the amino acid. More preferably, it is bonded to a group selected from H and OH (for example, OH of a carboxy group and H of a hydroxy group and a mercapto group), and is bonded to a OH of a carboxy group. Is more preferable.
　ホウ素クラスター（又はリンカー構造）が、カルボキシ基との間で形成し得る結合としては、例えば、アミド結合（－ＣＯＮＨ－）、エステル結合（－ＣＯＯ－）、チオエステル結合（－ＣＯＳ－）等が挙げられ、好ましくは、アミド結合である。ヒドロキシ基との間で形成し得る結合としては、エステル結合（－ＯＣＯ－）、ウレタン結合（－ＯＣＯＮＨ－）等が挙げられる。メルカプト基との間で形成し得る結合としては、チオエステル結合（－ＳＣＯ－、－ＣＯＳ－、ジスルフィド結合（－ＳＳ－）等が挙げられる。 Examples of the bond that the boron cluster (or linker structure) can form with the carboxy group include an amide bond (—CONH—), an ester bond (—COO—), a thioester bond (—COS—), and the like. Preferably, it is an amide bond. Examples of the bond that can be formed with the hydroxy group include an ester bond (—OCO—) and a urethane bond (—OCONH—). Examples of the bond that can be formed with the mercapto group include a thioester bond (—SCO—, —COS—, disulfide bond (—SS—) and the like.
　ペプチドのアミノ酸残基数に対する、ホウ素クラスターの結合数の比は、好ましくは、０．１以上であり、より好ましくは、０．２以上であり、さらに好ましくは、０．３以上である。 The ratio of the number of bonds of the boron cluster to the number of amino acid residues of the peptide is preferably 0.1 or more, more preferably 0.2 or more, and further preferably 0.3 or more.
　ホウ素クラスターからなる１価の基は、好ましくは、式（Ｂ１）～式（Ｂ１５）： The monovalent group consisting of boron clusters is preferably formula (B1) to formula (B15):
Figure JPOXMLDOC01-appb-C000015
Figure JPOXMLDOC01-appb-C000015
［式中、＊は、アミノ酸の側鎖又はリンカー構造との結合部位を示し、その他の記号は上記定義の通りである。］
の何れかで表される基であり、特に好ましくは、式（Ｂ１）で表される基である。 [Wherein, * represents a binding site with an amino acid side chain or linker structure, and other symbols are as defined above]. ]
And is particularly preferably a group represented by the formula (B1).
　本明細書における「リンカー構造」は、ホウ素クラスターとアミノ酸の側鎖とを連結させることができる薬学的に許容される構造であれば特に限定されるものではない。例えば、炭素原子、酸素原子、硫黄原子及び窒素原子から選ばれる主鎖構成原子（好ましくは、１～２００個、より好ましくは、１～３０個）からなる２価の基が挙げられる。 The “linker structure” in the present specification is not particularly limited as long as it is a pharmaceutically acceptable structure capable of linking a boron cluster and an amino acid side chain. Examples thereof include a divalent group consisting of main chain constituent atoms selected from carbon atoms, oxygen atoms, sulfur atoms and nitrogen atoms (preferably 1 to 200, more preferably 1 to 30).
　ある実施形態では、「リンカー構造」は、好ましくは、下記式（Ｌ）で表される２価の基である： In an embodiment, the “linker structure” is preferably a divalent group represented by the following formula (L):
Figure JPOXMLDOC01-appb-C000016
Figure JPOXMLDOC01-appb-C000016
［式中、＊１はアミノ酸の側鎖との結合部位であり、＊２はホウ素クラスターとの結合部位であり、Ｐ１及びＱ１は、それぞれ独立して 、－Ｏ－、－Ｓ－、－ＮＨ－、－ＳＯ２－、－ＣＯ－、－ＮＨＣＯ－、－ＣＯＮＨ－、－ＯＣＯ－、－ＣＯＯ－、－ＳＣＯ－、－ＣＯＳ－、－ＮＨＣＯＮＨ－、－ＮＨＣＳＮＨ－、－ＯＣＯＮＨ－、－ＮＨＣＯＯ－、－ＳＳ－、－ＮＨＣＯＳ－、－ＳＣＯＮＨ－、－ＮＨＣＨ２ＣＯＮＨ－、－ＮＨＣＯＣＨ２ＮＨ－、－ＮＨＣＨ２ＣＯＯ－、－ＯＣＯＣＨ２ＮＨ－、－ＯＣＨ２ＣＯＮＨ－、－ＮＨＣＯＣＨ２Ｏ－、－ＯＣＨ２ＣＯＯ－、－ＯＣＯＣＨ２Ｏ－、－ＳＣＨ２ＣＯＮＨ－、－ＮＨＣＯＣＨ２Ｓ－、－ＳＣＨ２ＣＯＯ－、－ＯＣＯＣＨ２Ｓ－、－Ｎ（－ＣＯＣＨ２－）（－ＣＯ－）ＣＨＳ－、－ＳＣＨ（－ＣＨ２ＣＯ－）（－ＣＯ－）Ｎ－、－Ｎ（－Ｎ＝Ｎ－）（－ＣＨ＝）Ｃ－、－Ｃ（－Ｎ＝Ｎ－）（＝ＣＨ－）Ｎ －、－ＮＨ－Ｎ＝ＣＨ－、－ＣＨ＝Ｎ－ＮＨ－、－Ｐｈ－、－ＯＰｈＯ－、－ＯＰｈ－、－ＰｈＯ－、－ＳＰｈＳ－、－ＳＰｈ－、－ＰｈＳ－、－ＮＨＰｈＮＨ－、－ＮＨＰｈ－、－ＰｈＮＨ－、－複素環－、－Ｏ－複素環－Ｏ－、－Ｏ－複素環－、－複素環－Ｏ－、－Ｓ－複素環－Ｓ－、－Ｓ－複素環－、－複素環－Ｓ－、－ＮＨ－複素環－ＮＨ－、－ＮＨ－複素環－、－複素環－ＮＨ－等（ここで、Ｐｈは、１，２－フェニレン、１，３－フェニレン又は１，４－フェニレン等を示す。）を示し、Ｌは、エチレングリコール、オリゴエチレングリコール、ポリエチレングリコール、オリゴエーテル、ポリエーテル等の２価の鎖状基、鎖状アルキレン基、環状アルキレン基等のアルキレン基等を示す。］。Ｌで表される２価の鎖状基及びアルキレン基は、それぞれ、その鎖状又は環状構造内に複素環やフェニレン（１，２－フェニレン、１，３－フェニレン又は１，４－フェニレン等）を含んでいてもよい。 [Wherein * 1 is a binding site with an amino acid side chain, * 2 is a binding site with a boron cluster, and P 1 and Q 1 are each independently —O—, —S—, —NH—, —SO 2 —, —CO—, —NHCO—, —CONH—, —OCO—, —COO—, —SCO—, —COS—, —NHCONH—, —NHCSNH—, —OCONH—, — NHCOO -, - SS -, - NHCOS -, - SCONH -, - NHCH 2 CONH -, - NHCOCH 2 NH -, - NHCH 2 COO -, - OCOCH 2 NH -, - OCH 2 CONH -, - NHCOCH 2 O- , -OCH 2 COO -, - OCOCH 2 O -, - SCH 2 CONH -, - NHCOCH 2 S -, - SCH 2 COO -, - OCOCH 2 S -, - N (-COCH 2 -) (- C O-) CHS-, -SCH (-CH 2 CO-) (-CO-) N-, -N (-N = N-) (-CH =) C-, -C (-N = N-) ( = CH-) N-, -NH-N = CH-, -CH = N-NH-, -Ph-, -OPhO-, -OPh-, -PhO-, -SPhS-, -SPh-, -PhS- , -NHPhNH-, -NHPh-, -PhNH-, -heterocycle-, -O-heterocycle-O-, -O-heterocycle-, -heterocycle-O-, -S-heterocycle-S-, -S-heterocycle-, -heterocycle-S-, -NH-heterocycle-NH-, -NH-heterocycle-, -heterocycle-NH- and the like (where Ph is 1,2-phenylene, 1, 3-phenylene, 1,4-phenylene, etc.), and L is ethylene glycol, oligoethylene glycol, polyethylene glycol, oligoacetate Le, a divalent chain group such as polyether, chain alkylene group, and the like alkylene group such as a cyclic alkylene group shown. ]. The divalent chain group and alkylene group represented by L are each a heterocycle or phenylene (1,2-phenylene, 1,3-phenylene, 1,4-phenylene, etc.) within the chain or cyclic structure. May be included.
　このようなリンカー構造は、ホウ素クラスターとアミノ酸の側鎖を連結させるために、種々の公知反応、公知のリンカー構造を選択して導入することができる。このようなリンカー構造は、広く知られており、例えば、Ｓ．Ｍａｎａｂｅ ａｎｄ　Ｍ．Ｙｏｋｏｙａｍａ，Ｄｒｕｇ　Ｄｅｌｉｖｅｒｙ　Ｓｙｓｔｅｍ，３０－３：２４７－２５０（２０１５）、Ｎ．Ｊａｉｎ　ｅｔ　ａｌ．，Ｐｈａｒｍ　Ｒｅｓ，３２：３２５６－３５４０（２０１５）、Ｊ．Ｒ．ＭｃＣｏｍｂｓ　ｅｔ　ａｌ．，Ｔｈｅ　ＡＡＰＳ　Ｊｏｕｒｎａｌ，１７（２）:３３９－３５１（Ｍａｒｃｈ　２０１５）、Ｊ．Ｋｈａｎｄａｒｅ　ｅｔ　ａｌ．，Ｐｒｏｇ．Ｐｏｌｙｍ．Ｓｃｉ．，３１:３５９-３９７（２００６）等の文献に記載されている。 Such a linker structure can be introduced by selecting various known reactions and known linker structures in order to link boron clusters and amino acid side chains. Such linker structures are well known and are described, for example, in S.I. Manabeand M.M. Yokoyama, Drug Delivery System, 30-3: 247-250 (2015), N.I. Jain et al. Pharm Res, 32: 3256-3540 (2015), J. Am. R. McCombs et al. The AAPS Journal, 17 (2): 339-351 (March 2015), J.A. Khandare et al. , Prog. Polym. Sci. 31: 359-397 (2006).
　別の実施形態では、「リンカー構造」は、好ましくは、下記式（Ｌ’）で表される２価の基である： In another embodiment, the “linker structure” is preferably a divalent group represented by the following formula (L ′):
Figure JPOXMLDOC01-appb-C000017
Figure JPOXMLDOC01-appb-C000017
［式中、
　Ｒａは、それぞれ独立して、水素原子、炭素数が１～６のアルキル基で置換されていてもよいカルボキシ基、炭素数が１～６のアルキル基で置換されていてもよいヒドロキシ基、炭素数が１～６のアルキル基で置換されていてもよいメルカプト基、炭素数が６～１４のアリール基、又は炭素数が１～６のアルキル基を示し；
　Ｐ２及びＱ２は、それぞれ独立して、－Ｏ－、－Ｓ－、－ＮＨ－、－ＳＯ２－、－ＣＯ－、－ＮＨＣＯ－、－ＣＯＮＨ－、－ＯＣＯ－、－ＣＯＯ－、－ＳＣＯ－、－ＣＯＳ－、－ＮＨＣＯＮＨ－、－ＮＨＣＳＮＨ－、－ＯＣＯＮＨ－、－ＮＨＣＯＯ－、－ＳＳ－、－Ｐｈ－、－ＯＰｈＯ－、－ＯＰｈ－、－ＰｈＯ－、－ＳＰｈＳ－、－ＳＰｈ－、－ＰｈＳ－、－ＮＨＰｈＮＨ－、－ＮＨＰｈ－、－ＰｈＮＨ－、－複素環－、－Ｏ－複素環－Ｏ－、－Ｏ－複素環－、－複素環－Ｏ－、－Ｓ－複素環－Ｓ－、－Ｓ－複素環－、－複素環－Ｓ－、－ＮＨ－複素環－ＮＨ－、－ＮＨ－複素環－、－複素環－ＮＨ－等（ここで、Ｐｈは、１，２－フェニレン、１，３－フェニレン又は１，４－フェニレンを示す。）を示し；
　ｆは、それぞれ独立して、１～２０の整数を示し；
　ｇは、０～２０（好ましくは０～１０、より好ましくは０～６）の整数を示し、その他の記号は上記定義の通りである。］。 [Where:
Each R a is independently a hydrogen atom, a carboxy group optionally substituted with an alkyl group having 1 to 6 carbon atoms, a hydroxy group optionally substituted with an alkyl group having 1 to 6 carbon atoms, A mercapto group optionally substituted with an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 14 carbon atoms, or an alkyl group having 1 to 6 carbon atoms;
P 2 and Q 2 are each independently —O—, —S—, —NH—, —SO 2 —, —CO—, —NHCO—, —CONH—, —OCO—, —COO—, — SCO—, —COS—, —NHCONH—, —NHCSNH—, —OCONH—, —NHCOO—, —SS—, —Ph—, —OPhO—, —OPh—, —PhO—, —SPhS—, —SPh— , -PhS-, -NHPhNH-, -NHPh-, -PhNH-, -heterocycle-, -O-heterocycle-O-, -O-heterocycle-, -heterocycle-O-, -S-heterocycle -S-, -S-heterocycle-, -heterocycle-S-, -NH-heterocycle-NH-, -NH-heterocycle-, -heterocycle-NH- and the like (where Ph is 1, Represents 2-phenylene, 1,3-phenylene or 1,4-phenylene);
f independently represents an integer of 1 to 20;
g represents an integer of 0 to 20 (preferably 0 to 10, more preferably 0 to 6), and other symbols are as defined above. ].
　このようなリンカー構造は、ホウ素クラスターとアミノ酸の側鎖を連結させるために、種々の公知反応、公知のリンカー構造を選択して導入することができる。 Such a linker structure can be introduced by selecting various known reactions and known linker structures in order to link boron clusters and amino acid side chains.
　本明細書における「複素環」としては、環構成原子として炭素原子以外に酸素原子、硫黄原子及び窒素原子からなる群から選ばれる１～４個のヘテロ原子を含有する環であって、１以上のオキソ基で置換されていてもよい複素環が挙げられ、具体的には、アゼチジン、ピロリジン、ピペリジン、モルホリン、チオモルホリン、ピペラジン、オキサゾリジン、チアゾリジン、ジヒドロチオピラン、イミダゾリジン、オキサゾリン、チアゾリン、イミダゾリン、ジオキソール、ジオキソラン、ジヒドロオキサジアゾール、ピラン、ジヒドロピラン、テトラヒドロピラン、チオピラン、ジヒドロチオピラン、テトラヒドロチオピラン、テトラヒドロフラン、オキセタン、ピラゾリジン、ピラゾリン、テトラヒドロピリミジン、ジヒドロトリアゾール、アゼパン、ジヒドロピリジン、テトラヒドロピリジン、ジオキソピロリジン等の３～８員の非芳香族複素環；フラン、チオフェン、ピリジン、ピリミジン、ピリダジン、ピラジン、ピロール、イミダゾール、ピラゾール、チアゾール、イソチアゾール、オキサゾール、イソオキサゾール、オキサジアゾール、チアジアゾール、トリアゾール、テトラゾール、トリアジン等の５又は６員の芳香族複素環等が挙げられる。 As used herein, the term “heterocycle” refers to a ring containing 1 to 4 heteroatoms selected from the group consisting of an oxygen atom, a sulfur atom and a nitrogen atom in addition to a carbon atom as a ring constituent atom. Specific examples include hetetidine, pyrrolidine, piperidine, morpholine, thiomorpholine, piperazine, oxazolidine, thiazolidine, dihydrothiopyran, imidazolidine, oxazoline, thiazoline, imidazoline. , Dioxol, dioxolane, dihydrooxadiazole, pyran, dihydropyran, tetrahydropyran, thiopyran, dihydrothiopyran, tetrahydrothiopyran, tetrahydrofuran, oxetane, pyrazolidine, pyrazoline, tetrahydropyrimidine, dihydrotriazol , Azepane, dihydropyridine, tetrahydropyridine, dioxopyrrolidine, etc. 3-8 membered non-aromatic heterocycles; furan, thiophene, pyridine, pyrimidine, pyridazine, pyrazine, pyrrole, imidazole, pyrazole, thiazole, isothiazole, oxazole, iso Examples thereof include 5- or 6-membered aromatic heterocycles such as oxazole, oxadiazole, thiadiazole, triazole, tetrazole, and triazine.
　本明細書における「炭素数が１～６のアルキル基」とは、炭素数が１～６の直鎖、分枝鎖又は環状の飽和炭化水素基を意味し、例えば、メチル、エチル、プロピル、イソプロピル、ブチル、イソブチル、ｓｅｃ－ブチル、ｔｅｒｔ－ブチル、ペンチル、シクロペンチル、シクロヘキシル等が挙げられる。 As used herein, “an alkyl group having 1 to 6 carbon atoms” means a linear, branched or cyclic saturated hydrocarbon group having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, Examples include isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, cyclopentyl, cyclohexyl and the like.
　本明細書における「炭素数が６～１４のアリール基」とは、炭素数が６～１４の環状の芳香族炭化水素基を意味し、例えば、フェニル、１－ナフチル、２－ナフチル等が挙げられる。 In the present specification, the “aryl group having 6 to 14 carbon atoms” means a cyclic aromatic hydrocarbon group having 6 to 14 carbon atoms, and examples thereof include phenyl, 1-naphthyl, 2-naphthyl and the like. It is done.
　Ｒａは、それぞれ、好ましくは、水素原子である。 Each R a is preferably a hydrogen atom.
　Ｐ１及びＰ２は、それぞれ、アミノ酸の側鎖における結合対象がＯ又はＳ（例えば、ヒドロキシ基又はメルカプト基）の場合は、－ＣＯ－、－ＣＯＮＨ－又は－ＣＯＯ－であることが好ましく、アミノ酸の側鎖における結合対象がＣＯ（例えば、カルボキシ基）の場合は、－Ｏ－、－ＮＨ－又は－Ｓ－であることが好ましい。 P 1 and P 2 are each preferably —CO—, —CONH— or —COO— when the binding target in the side chain of the amino acid is O or S (for example, a hydroxy group or a mercapto group), When the binding target in the side chain of an amino acid is CO (for example, a carboxy group), it is preferably —O—, —NH— or —S—.
　Ｑ２は、好ましくは、それぞれ独立して、－ＮＨＣＯ－、－ＣＯＮＨ－、－Ｏ－、－Ｓ－、－ＳＳ－又は－複素環－である。 Q 2 is preferably each independently —NHCO—, —CONH—, —O—, —S—, —SS— or —heterocycle—.
　本明細書において「ポリエーテル」とは、主鎖にエーテル結合を有する直鎖又は分枝鎖の重合体を意味し、好ましくは、主にポリアルキレングリコール単位（好ましくはポリエチレングリコール（ＰＥＧ）単位）を含む直鎖又は分枝鎖の水溶性重合体である。ポリアルキレングリコール単位は、ポリエーテル中に６０重量％以上含まれていることが好ましく、８０重量％以上含まれていることがより好ましく、９０重量％以上含まれていることがさらに好ましい。ポリエーテルの主鎖の末端部分又は鎖状部分の原子は、酸素原子及び炭素原子以外の原子（例えば、硫黄原子、窒素原子等）であってもよい。 In the present specification, the “polyether” means a linear or branched polymer having an ether bond in the main chain, preferably a polyalkylene glycol unit (preferably a polyethylene glycol (PEG) unit). Is a linear or branched water-soluble polymer. The polyalkylene glycol unit is preferably contained in the polyether in an amount of 60% by weight or more, more preferably 80% by weight or more, and further preferably 90% by weight or more. The atoms of the terminal part or the chain part of the main chain of the polyether may be atoms other than oxygen atoms and carbon atoms (for example, sulfur atoms, nitrogen atoms, etc.).
　本明細書における「ポリエーテル」の数平均分子量（Ｍｎ）は、好ましくは、５０，０００未満であり、より好ましくは、３０，０００未満である。また、「ポリエーテル」の数平均分子量（Ｍｎ）は、好ましくは、１，０００以上であり、より好ましくは、１０，０００以上である。「ポリエーテル」の分子量分布（重量平均分子量（Ｍｗ）／数平均分子量（Ｍｎ））は、好ましくは、３以下であり、より好ましくは、２以下である。 The number average molecular weight (Mn) of “polyether” in the present specification is preferably less than 50,000, and more preferably less than 30,000. Further, the number average molecular weight (Mn) of the “polyether” is preferably 1,000 or more, more preferably 10,000 or more. The molecular weight distribution (weight average molecular weight (Mw) / number average molecular weight (Mn)) of the “polyether” is preferably 3 or less, more preferably 2 or less.
　本発明のペプチド化合物において、ポリエーテルは、ペプチドの構成アミノ酸の活性基（例えば、ペプチド化合物の構成アミノ酸の側鎖のカルボキシ基、ヒドロキシ基及びメルカプト基、並びにペプチド化合物のＮ末端のアミノ基及びＣ末端のカルボキシ基）から選ばれる１個以上の基に結合していることが好ましく、特にペプチド化合物のＮ末端のアミノ基若しくはＣ末端のカルボキシ基のいずれかに結合していることが好ましい。 In the peptide compound of the present invention, the polyether is an active group of a constituent amino acid of the peptide (for example, a carboxy group, a hydroxy group and a mercapto group on the side chain of the constituent amino acid of the peptide compound, and an amino group and C at the N terminal of the peptide compound. It is preferably bonded to one or more groups selected from (terminal carboxy group), and particularly preferably bonded to either the N-terminal amino group or the C-terminal carboxy group of the peptide compound.
　また、ポリエーテルは、ペプチドに１価の基（ポリエーテルの任意の１個のＨ又はＯＨを除いてなる基を意味し、以下において「ポリエーテルからなる１価の基」という。）として結合していてもよく、或いは多価の基（ポリエーテルの任意のＨ及びＯＨから選ばれる２個以上の基を除いてなる基を意味し、以下において「ポリエーテルからなるｃ価の基」（ｃは２以上の整数）という。）として複数のペプチドに結合している形態であってもよい。ポリエーテルからなる１価の基及びポリエーテルからなるｃ価の基は、それぞれ、好ましくは、ポリエーテル鎖の末端におけるカルボキシ基のＯＨ、並びにアミノ基、ヒドロキシ基及びメルカプト基のＨのうち１個又はｃ個を除いてなる基である。 The polyether is bonded to the peptide as a monovalent group (meaning a group formed by removing any one H or OH of the polyether, and hereinafter referred to as “monovalent group made of polyether”). Or a polyvalent group (meaning a group formed by removing two or more groups selected from any H and OH of a polyether, and hereinafter referred to as a “c-valent group comprising a polyether” ( c is an integer of 2 or more))) and may be bonded to a plurality of peptides. The monovalent group made of polyether and the c-valent group made of polyether are each preferably one of OH of the carboxy group and H of the amino group, hydroxy group and mercapto group at the end of the polyether chain. Or it is group which removes c pieces.
　ポリエーテルが、アミノ基、カルボキシ基、ヒドロキシ基及びメルカプト基との間で形成し得る結合としては、それぞれ、ホウ素クラスター（又はリンカー構造）が形成し得る結合として上記で挙げたものと同様のものが挙げられる。 As the bond that the polyether can form with an amino group, a carboxy group, a hydroxy group, and a mercapto group, the same bonds as those listed above as the bonds that the boron cluster (or linker structure) can form respectively. Is mentioned.
　上記「ポリエーテルからなる１価の基」の具体例としては、下記式（Ｐ１）～（Ｐ３）：  Specific examples of the above-mentioned “monovalent group consisting of polyether” include the following formulas (P1) to (P3):
Figure JPOXMLDOC01-appb-C000018
Figure JPOXMLDOC01-appb-C000018
［式中、
　Ｚは、－Ｏ－、－ＮＨ－、－Ｓ－、－ＣＯ－、－ＣＯＮＨ－、－ＣＯＯ－、－ＮＨＣＯ－、－ＣＯＮＨ－、－ＯＣＯ－、－ＣＯＯ－、－ＳＣＯ－、－ＣＯＳ－、－ＮＨＣＯＮＨ－、－ＮＨＣＳＮＨ－、－ＯＣＯＮＨ－、－ＮＨＣＯＯ－又は－ＳＳ－を示し、
　Ｒｄ及びＲｅは、それぞれ独立して水素原子又は炭素数が１～６のアルキル基を示し、
　ｓは、それぞれ独立して１～１０（好ましくは１～４）の整数であり、
　ｔは、それぞれ独立して２～１０（好ましくは２～４）の整数であり、
　ｕは、それぞれ独立して１以上の整数であり、
　＊＊＊は、結合部位を示す。］
で表される基が挙げられる。中でも好ましくは、上記式（Ｐ１）で表される基である。 [Where:
Z represents —O—, —NH—, —S—, —CO—, —CONH—, —COO—, —NHCO—, —CONH—, —OCO—, —COO—, —SCO—, —COS—. , -NHCONH-, -NHCSNH-, -OCONH-, -NHCOO- or -SS-
R d and R e each independently represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms,
s each independently represents an integer of 1 to 10 (preferably 1 to 4),
t is each independently an integer of 2 to 10 (preferably 2 to 4);
u is each independently an integer of 1 or more,
*** indicates a binding site. ]
The group represented by these is mentioned. Among them, a group represented by the above formula (P1) is preferable.
　ｃが２の場合における上記「ポリエーテルからなるｃ価の基」の具体例は、式（Ｐ４）又は（Ｐ５）： Specific examples of the above-mentioned “c-valent group consisting of polyether” when c is 2 are represented by the formula (P4) or (P5):
Figure JPOXMLDOC01-appb-C000019
Figure JPOXMLDOC01-appb-C000019
［式中、各記号は上記定義の通りである。］
で表される基が挙げられる。 [Wherein the symbols are as defined above. ]
The group represented by these is mentioned.
　Ｒｄは、好ましくは、水素原子である。Ｒｅは、好ましくは、炭素数が１～６のアルキル基であり、より好ましくはメチルである。 R d is preferably a hydrogen atom. R e is preferably an alkyl group having 1 to 6 carbon atoms, more preferably methyl.
　ｕの合計は、好ましくは、１，０００以下の整数であり、より好ましくは、６００以下の整数である。 The total of u is preferably an integer of 1,000 or less, more preferably an integer of 600 or less.
　Ｚは、結合対象がＯ、Ｎ、ＮＨ又はＳ（例えばヒドロキシ基、アミノ基又はメルカプト基）の場合、－ＣＯ－、－ＣＯＮＨ－又は－ＣＯＯ－であることが好ましく、結合対象がＣＯ（例えばカルボキシ基）の場合、－Ｏ－、－ＮＨ－又は－Ｓ－であることが好ましい。 Z is preferably —CO—, —CONH— or —COO— when the binding target is O, N, NH or S (for example, a hydroxy group, an amino group or a mercapto group), and the binding target is CO (for example, In the case of a carboxy group, it is preferably —O—, —NH— or —S—.
　ある実施形態において、好ましくは、ペプチドの構成アミノ酸の活性基の少なくとも１個が、ポリエーテルからなる１価の基で置換されている。より好ましくは、ペプチド化合物のＣ末端のカルボキシ基及び／又はペプチド化合物のＮ末端のアミノ基が、ポリエーテルからなる１価の基で置換されている。さらに好ましくは、ペプチド化合物のＣ末端のカルボキシ基及び／又はペプチド化合物のＮ末端のアミノ基が、式（Ｐ１）（式中、式中、Ｚは、置換対象がカルボキシ基の場合－Ｏ－、－ＮＨ－又は－Ｓ－を、置換対象がアミノ基の場合－ＣＯ－、－ＣＯＮＨ－又は－ＣＯＯ－を示し、その他の記号は上記定義の通りである。）で表される基で置換されている。 In one embodiment, preferably, at least one of the active groups of the constituent amino acids of the peptide is substituted with a monovalent group consisting of a polyether. More preferably, the C-terminal carboxy group of the peptide compound and / or the N-terminal amino group of the peptide compound are substituted with a monovalent group comprising a polyether. More preferably, the carboxy group at the C-terminal of the peptide compound and / or the amino group at the N-terminal of the peptide compound are represented by the formula (P1) (wherein Z is —O— when the substitution target is a carboxy group, When —NH— or —S— is substituted with an amino group, it represents —CO—, —CONH— or —COO—, and the other symbols are as defined above. ing.
　ある実施形態では、ホウ素クラスター（又はリンカー構造）が、側鎖に結合したアミノ酸残基は、下記式（１）で表される。 In an embodiment, an amino acid residue having a boron cluster (or linker structure) bonded to a side chain is represented by the following formula (1).
Figure JPOXMLDOC01-appb-C000020
Figure JPOXMLDOC01-appb-C000020
［式中、各記号は上記の定義の通りである。］ [Wherein each symbol is as defined above. ]
　本明細書において「α－アミノ酸」とは、１個の炭素原子（α－炭素とよばれる）に１個のカルボキシル基と１個のアミノ基の両方が結合しているアミノ酸を意味する。α－アミノ酸はＤ型、Ｌ型又はそれらの混合物であってもよい。 In the present specification, “α-amino acid” means an amino acid in which one carboxyl group and one amino group are bonded to one carbon atom (referred to as α-carbon). The α-amino acid may be D-form, L-form or a mixture thereof.
　本明細書において「α－アミノ酸の側鎖」とは、α－アミノ酸をＲ３ＣＨ（ＮＨ２）ＣＯＯＨで表したときに、Ｒ３に相当する基をいう。具体的には、例えば、α－アミノ酸がグリシンの場合は水素原子、アラニンの場合はメチル、バリンの場合はイソプロピル、ロイシンの場合はイソブチル、イソロイシンの場合はｓｅｃ－ブチル、セリンの場合はヒドロキシメチル、トレオニンの場合は１－ヒドロキシエチル、システインの場合はメルカプトメチル、メチオニンの場合は２－（メチルチオ）エチル、アスパラギンの場合はカルバモイルメチル、グルタミンの場合は２－カルバモイルエチル、フェニルアラニンの場合はベンジル、チロシンの場合はｐ－ヒドロキシベンジル、アスパラギン酸の場合はカルボキシメチル、グルタミン酸の場合は２－カルボキシエチル、ホモセリンの場合は２－ヒドロキシエチル、ノルロイシンの場合はブチル、ノルバリンの場合はプロピル、チロニンの場合は４－｛（４－ヒドロキシフェニル）オキシ｝フェニルメチル、シトルリンの場合は３－（カルバモイルアミノ）プロパン、α-アミノ酪酸の場合はエチル、ホモシステインの場合は２－メルカプトエチル、ペニシラミンの場合は１－メルカプト－１－メチルエチルである。 In the present specification, the “side chain of α-amino acid” refers to a group corresponding to R 3 when the α-amino acid is represented by R 3 CH (NH 2 ) COOH. Specifically, for example, when the α-amino acid is glycine, it is a hydrogen atom, when it is alanine, it is methyl, when it is valine, it is isopropyl, when it is leucine, it is isobutyl, when it is isoleucine, it is sec-butyl, and when it is serine, it is hydroxymethyl. 1-hydroxyethyl for threonine, mercaptomethyl for cysteine, 2- (methylthio) ethyl for methionine, carbamoylmethyl for asparagine, 2-carbamoylethyl for glutamine, benzyl for phenylalanine, P-hydroxybenzyl for tyrosine; carboxymethyl for aspartic acid; 2-carboxyethyl for glutamic acid; 2-hydroxyethyl for homoserine; butyl for norleucine; propyl for norvaline; 4-{(4-hydroxyphenyl) oxy} phenylmethyl for citron, 3- (carbamoylamino) propane for citrulline, ethyl for α-aminobutyric acid, 2-mercaptoethyl, penicillamine for homocysteine In the case of 1-mercapto-1-methylethyl.
　Ｒ１のα－アミノ酸の側鎖は、それが結合する炭素原子及び該炭素原子に隣接する窒素原子と一緒になって、環を形成していてもよい。ここにおける環状部分としては、例えば、プロリンの環状部分（即ち、ピロリジン環）が挙げられる。ある実施形態では、Ｒ１のα－アミノ酸の側鎖は、それが結合する炭素原子及び該炭素原子に隣接する窒素原子と一緒になって環を形成しない。 The side chain of the α-amino acid of R 1 may be combined with the carbon atom to which it is bonded and the nitrogen atom adjacent to the carbon atom to form a ring. Examples of the cyclic moiety herein include a proline cyclic moiety (that is, a pyrrolidine ring). In certain embodiments, the side chain of the α-amino acid of R 1 does not form a ring with the carbon atom to which it is attached and the nitrogen atom adjacent to the carbon atom.
　Ｒ１の「α－アミノ酸」は、好ましくは、セリン、トレオニン、システイン、チロシン、アスパラギン酸、グルタミン酸、ホモセリン、ホモシステイン、ペニシラミン等の側鎖にカルボキシ基、ヒドロキシ基、メルカプト基等の活性基を有するα－アミノ酸であり；なかでも、グルタミン酸及びアスパラギン酸から選ばれることがより好ましく；グルタミン酸であることが特に好ましい。 The “α-amino acid” of R 1 preferably has an active group such as carboxy group, hydroxy group or mercapto group on the side chain of serine, threonine, cysteine, tyrosine, aspartic acid, glutamic acid, homoserine, homocysteine, penicillamine and the like. Among them, it is more preferable that the amino acid is selected from glutamic acid and aspartic acid; and particularly preferable is glutamic acid.
　Ｒ１における、ホウ素クラスターからなる１価の基又はそれがリンカー構造を介して結合する１価の基の置換数は、１～３であることが好ましく、１であることがより好ましい。 The number of substitution of a monovalent group consisting of boron clusters or a monovalent group to which it is bonded via a linker structure in R 1 is preferably 1 to 3, and more preferably 1.
　Ｒ１は、好ましくは、それぞれ独立して、ホウ素クラスターからなる１価の基（中でも好ましくは、式（Ｂ１）～式（Ｂ１５）の何れかで表される基）がリンカー構造を介して結合する１価の基で置換された、中性アミノ酸及び酸性アミノ酸から選ばれるα－アミノ酸の側鎖である。 Preferably, each R 1 is independently a monovalent group composed of a boron cluster (particularly preferably, a group represented by any one of the formulas (B1) to (B15)) is bonded via a linker structure. A side chain of an α-amino acid selected from neutral amino acids and acidic amino acids substituted with a monovalent group.
　Ｒ１は、より好ましくは、それぞれ独立して、式（Ｂ１）で表される基がリンカー構造を介して結合する１価の基で置換された、中性アミノ酸及び酸性アミノ酸から選ばれるα－アミノ酸の側鎖である。 More preferably, R 1 is independently α- selected from a neutral amino acid and an acidic amino acid, wherein each group represented by the formula (B1) is substituted with a monovalent group bonded via a linker structure. A side chain of an amino acid.
　Ｒ１は、さらに好ましくは、Ｒ１の「α－アミノ酸」が、グルタミン酸及びアスパラギン酸からなる群から選ばれる場合であって、それぞれ独立して、式（Ｒ１ａ）： R 1 is more preferably the case where the “α-amino acid” of R 1 is selected from the group consisting of glutamic acid and aspartic acid, and each independently represents formula (R1a):
Figure JPOXMLDOC01-appb-C000021
Figure JPOXMLDOC01-appb-C000021
［式中、Ｐ３は、－Ｏ－、－ＮＨ－又は－Ｓ－を示し、ｄは、１又は２を示し、＊＊は、α－アミノ酸のα－炭素との結合部位を示し、その他の記号は上記の定義の通りである。］
で表される基である。 [Wherein P 3 represents —O—, —NH— or —S—, d represents 1 or 2, ** represents a binding site to the α-carbon of an α-amino acid, and others The symbol is as defined above. ]
It is group represented by these.
　ある実施形態では、ペプチド化合物は、上記のホウ素クラスター（リンカー構造を含む）の結合、及びポリエーテル（例えば「ポリエーテルからなる１価の基」、「ポリエーテルからなるｃ価の基」）の結合に加えて、さらに、その他の置換基を有していてもよい。置換基を有する場合、１個以上の置換基が、アミノ酸の側鎖で置換していてもよく、中でも、アミノ酸の側鎖の活性基（例えば、カルボキシ基、ヒドロキシ基及びメルカプト基）のＨ又はＯＨと置換していることが好ましい。 In one embodiment, the peptide compound comprises a bond of the above boron cluster (including a linker structure) and a polyether (eg, “monovalent group composed of polyether”, “c-valent group composed of polyether”). In addition to the bond, it may further have other substituents. When it has a substituent, one or more substituents may be substituted with the side chain of the amino acid, and among them, the active group of the side chain of the amino acid (for example, carboxy group, hydroxy group and mercapto group) H or It is preferably substituted with OH.
　ホウ素クラスターをリンカー構造を介して導入する際の反応において、未反応によりホウ素クラスターが導入されず、リンカー構造又はその断片構造が残存する場合があるが、上記その他の置換基には、そのような残存した未反応の構造が含まれ得る。さらに、上記その他の置換基には、未反応の構造を当業者に公知の方法により不活性化した構造や、副反応により生成した構造、脱保護反応で脱離しなかった未脱保護基を含む構造等も含まれ得る。このように未反応により残存したリンカー構造又はその断片構造の末端は、多種多様な構造を取り得るため、特定の置換基に限定されるものではない。また、上記その他の置換基には、ペプチド化学で用いられる当業者に公知の修飾基等も含まれ得る。 In the reaction when introducing the boron cluster through the linker structure, the boron cluster may not be introduced due to unreacted, and the linker structure or a fragment structure thereof may remain. Residual unreacted structures may be included. In addition, the other substituents include structures obtained by inactivating unreacted structures by methods known to those skilled in the art, structures formed by side reactions, and undeprotected groups that were not removed by deprotection reactions. Structures and the like can also be included. Thus, the terminal of the linker structure or its fragment structure left unreacted can take a wide variety of structures and is not limited to a specific substituent. In addition, the above-mentioned other substituents may include modifying groups known to those skilled in the art used in peptide chemistry.
　ある実施形態において、上記その他の置換基としては、例えば、導入するリンカー構造が上記式（Ｌ’）である場合、式（Ｌ’’）： In one embodiment, as the other substituent, for example, when the linker structure to be introduced is the formula (L ′), the formula (L ″):
Figure JPOXMLDOC01-appb-C000022
Figure JPOXMLDOC01-appb-C000022
［式中、Ａ１’は、公知のカップリング反応における未反応の官能基、それを公知の方法により不活性化した基、それらの副反応により生成した基、又は未脱保護基を含む基であり、ｇ’は０～１９の整数を示し、その他の記号は上記の定義の通りである。］
で表される基が挙げられる。上記未反応の官能基としては、メルカプト基、ピリジン－２－イルジスルファニル基、カルボキシ基又はその活性基（例えば、４-ニトロフェノールエステル、ペンタフルオロフェノールエステルのような活性エステルや、酸クロリドのような酸ハライド等）、アミノ基、クリックケミストリーによる環化反応のためのアジド（Ｎ３）基、シクロオクチン基、エチニル（ＨＣ≡Ｃ）基等が挙げられるが、これらに限定されない。 [In the formula, A 1 ′ represents an unreacted functional group in a known coupling reaction, a group inactivated by a known method, a group generated by a side reaction thereof, or a group containing an undeprotected group. And g ′ represents an integer of 0 to 19, and other symbols are as defined above. ]
The group represented by these is mentioned. Examples of the unreacted functional group include a mercapto group, a pyridin-2-yldisulfanyl group, a carboxy group or an active group thereof (for example, an active ester such as 4-nitrophenol ester and pentafluorophenol ester, and an acid chloride). Non-acid halides), amino groups, azide (N 3 ) groups for cyclization reactions by click chemistry, cyclooctyne groups, ethynyl (HC≡C) groups, and the like.
　別の実施形態において、上記その他の置換基としては、例えば、下記（１）～（５）からなる置換基Ａ群から選ばれる置換基を挙げることができるが、これらに限定されない；置換基Ａ群：
（１）下記置換基（ａ）～（ｌ）から選ばれる１～５個の置換基で、それぞれ、アルキル、アルケニル、アルキニル又はアリール上が置換されていてもよい下記置換基（Ａ）～（ＡＡ）：
（Ａ）炭素数が１～６のアルキル基、（Ｂ）炭素数が２～６のアルケニル基、（Ｃ）炭素数が２～６のアルキニル基、（Ｄ）炭素数が６～１４のアリール基、（Ｅ）炭素数が２～７のアルキルカルボニル基、（Ｆ）炭素数が１～６のアルキル基で置換されていてもよいカルボキシ基、（Ｇ）炭素数が２～６のアルケニル基で置換されていてもよいカルボキシ基、（Ｈ）炭素数が２～６のアルキニル基で置換されていてもよいカルボキシ基、（Ｉ）炭素数が６～１４のアリール基で置換されていてもよいカルボキシ基、（Ｊ）炭素数が１～６のアルキル基で置換されていてもよいカルバモイル基、（Ｋ）炭素数が２～６のアルケニル基で置換されていてもよいカルバモイル基、（Ｌ）炭素数が２～６のアルキニル基で置換されていてもよいカルバモイル基、（Ｍ）炭素数が６～１４のアリール基で置換されていてもよいカルバモイル基、（Ｎ）炭素数が１～６のアルキル基で置換されていてもよいヒドロキシ基、（Ｏ）炭素数が２～６のアルケニル基で置換されていてもよいヒドロキシ基、（Ｐ）炭素数が２～６のアルキニル基で置換されていてもよいヒドロキシ基、（Ｑ）炭素数が６～１４のアリール基で置換されていてもよいヒドロキシ基、（Ｒ）炭素数が１～６のアルキル基で置換されていてもよいアミノ基、（Ｓ）炭素数が２～６のアルケニル基で置換されていてもよいアミノ基、（Ｔ）炭素数が２～６のアルキニル基で置換されていてもよいアミノ基、（Ｕ）炭素数が６～１４のアリール基で置換されていてもよいアミノ基、（Ｖ）炭素数が１～６のアルキル基で置換されていてもよいメルカプト基、（Ｗ）炭素数が２～６のアルケニル基で置換されていてもよいメルカプト基、（Ｘ）炭素数が２～６のアルキニル基で置換されていてもよいメルカプト基、（Ｙ）炭素数が６～１４のアリール基で置換されていてもよいメルカプト基、（Ｚ）炭素数が１～６のアルキルスルホニル基、及び（ＡＡ）炭素数が１～６のアルキル基で置換されていてもよいスルホ基；
（ａ）置換されていてもよい炭素数が６～１４のアリール基、（ｂ）ホルミル基、（ｃ）置換されていてもよい炭素数が２～７のアルキルカルボニル基、（ｄ）置換されていてもよい炭素数が１～６のアルキル基、置換されていてもよい炭素数が２～６のアルケニル基、置換されていてもよい炭素数が２～６のアルキニル基、及び置換されていてもよい炭素数が６～１４のアリール基から選ばれる置換基で置換されていてもよいカルボキシ基、（ｅ）置換されていてもよい炭素数が１～６のアルキル基、置換されていてもよい炭素数が２～６のアルケニル基、置換されていてもよい炭素数が２～６のアルキニル基、及び置換されていてもよい炭素数が６～１４のアリール基から選ばれる置換基で置換されていてもよいカルバモイル基、（ｆ）置換されていてもよい炭素数が１～６のアルキル基、置換されていてもよい炭素数が２～６のアルケニル基、置換されていてもよい炭素数が２～６のアルキニル基、及び置換されていてもよい炭素数が６～１４のアリール基から選ばれる置換基で置換されていてもよいヒドロキシ基、（ｇ）置換されていてもよい炭素数が１～６のアルキル基、置換されていてもよい炭素数が２～６のアルケニル基、置換されていてもよい炭素数が２～６のアルキニル基、及び置換されていてもよい炭素数が６～１４のアリール基から選ばれる置換基で置換されていてもよいメルカプト基、（ｈ）置換されていてもよい炭素数が１～６のアルキルスルホニル基、（ｉ）置換されていてもよい炭素数が１～６のアルキル基で置換されていてもよいスルホ基、（ｊ）ハロゲン原子、（ｋ）シアノ基、（ｌ）ニトロ基；
（２）ホルミル基
（３）ハロゲン原子、
（４）シアノ基、及び
（５）ニトロ基。 In another embodiment, examples of the other substituents include, but are not limited to, a substituent selected from the group of substituent A consisting of the following (1) to (5); group:
(1) 1 to 5 substituents selected from the following substituents (a) to (l), which may be substituted on alkyl, alkenyl, alkynyl or aryl, respectively, AA):
(A) an alkyl group having 1 to 6 carbon atoms, (B) an alkenyl group having 2 to 6 carbon atoms, (C) an alkynyl group having 2 to 6 carbon atoms, (D) an aryl having 6 to 14 carbon atoms (E) an alkylcarbonyl group having 2 to 7 carbon atoms, (F) a carboxy group optionally substituted with an alkyl group having 1 to 6 carbon atoms, and (G) an alkenyl group having 2 to 6 carbon atoms. A carboxy group optionally substituted with (H) a carboxy group optionally substituted with an alkynyl group having 2 to 6 carbon atoms, or (I) an optionally substituted aryl group with 6 to 14 carbon atoms. A good carboxy group, (J) a carbamoyl group optionally substituted with an alkyl group having 1 to 6 carbon atoms, (K) a carbamoyl group optionally substituted with an alkenyl group having 2 to 6 carbon atoms, (L ) Cal which may be substituted with an alkynyl group having 2 to 6 carbon atoms A moyl group, (M) a carbamoyl group optionally substituted with an aryl group having 6 to 14 carbon atoms, (N) a hydroxy group optionally substituted with an alkyl group having 1 to 6 carbon atoms, (O) A hydroxy group that may be substituted with an alkenyl group having 2 to 6 carbon atoms, (P) a hydroxy group that may be substituted with an alkynyl group having 2 to 6 carbon atoms, and (Q) a carbon number of 6 to 14 (R) an amino group optionally substituted with an alkyl group having 1 to 6 carbon atoms, and (S) an alkenyl group having 2 to 6 carbon atoms. An amino group which may be substituted, (T) an amino group which may be substituted with an alkynyl group having 2 to 6 carbon atoms, and (U) an amino group which may be substituted with an aryl group having 6 to 14 carbon atoms (V) substituted with an alkyl group having 1 to 6 carbon atoms An optional mercapto group, (W) a mercapto group optionally substituted with an alkenyl group having 2 to 6 carbon atoms, (X) a mercapto group optionally substituted with an alkynyl group having 2 to 6 carbon atoms, (Y) a mercapto group optionally substituted with an aryl group having 6 to 14 carbon atoms, (Z) an alkylsulfonyl group having 1 to 6 carbon atoms, and (AA) an alkyl group having 1 to 6 carbon atoms. An optionally substituted sulfo group;
(A) an optionally substituted aryl group having 6 to 14 carbon atoms, (b) a formyl group, (c) an optionally substituted alkylcarbonyl group having 2 to 7 carbon atoms, (d) a substituted group An optionally substituted alkyl group having 1 to 6 carbon atoms, an optionally substituted alkenyl group having 2 to 6 carbon atoms, an optionally substituted alkynyl group having 2 to 6 carbon atoms, and a substituted group. A carboxy group which may be substituted with a substituent selected from aryl groups having 6 to 14 carbon atoms, (e) an alkyl group having 1 to 6 carbon atoms which may be substituted, A substituent selected from an alkenyl group having 2 to 6 carbon atoms, an alkynyl group having 2 to 6 carbon atoms that may be substituted, and an aryl group having 6 to 14 carbon atoms that may be substituted; An optionally substituted carbamoyl group, (f) An alkyl group having 1 to 6 carbon atoms which may be substituted, an alkenyl group having 2 to 6 carbon atoms which may be substituted, an alkynyl group having 2 to 6 carbon atoms which may be substituted, and a substituent An optionally substituted hydroxy group optionally substituted with a substituent selected from aryl groups having 6 to 14 carbon atoms, (g) an optionally substituted alkyl group having 1 to 6 carbon atoms, A substituent selected from an alkenyl group having 2 to 6 carbon atoms, an alkynyl group having 2 to 6 carbon atoms that may be substituted, and an aryl group having 6 to 14 carbon atoms that may be substituted An optionally substituted mercapto group, (h) an optionally substituted alkylsulfonyl group having 1 to 6 carbon atoms, and (i) an optionally substituted alkyl group having 1 to 6 carbon atoms. An optionally substituted sulfo group, (j Halogen atom, (k) cyano, (l) a nitro group;
(2) formyl group (3) halogen atom,
(4) a cyano group, and (5) a nitro group.
　置換基Ａ群は、好ましくは、上記置換基（ａ）～（ｌ）から選ばれる１～５個の置換基で、それぞれ、アルキル上が置換されていてもよい上記置換基（Ａ）、（Ｅ）、（Ｆ）、（Ｊ）、（Ｚ）、（Ｒ）、（Ｖ）、（Ｚ）及び（ＡＡ）である。 Substituent group A preferably has 1 to 5 substituents selected from the above substituents (a) to (l), and each of the above substituents (A), ( E), (F), (J), (Z), (R), (V), (Z) and (AA).
　置換基Ａ群は、より好ましくは、下記置換基（ｃ’）～（ｌ’）から選ばれる１～５個の置換基で、それぞれ、アルキル上が置換されていてもよい上記置換基（Ａ）、（Ｅ）、（Ｆ）、（Ｊ）、（Ｚ）、（Ｒ）、（Ｖ）、（Ｚ）及び（ＡＡ）である；
（ｃ’）置換されていてもよい炭素数が２～７のアルキルカルボニル基、（ｄ’）置換されていてもよい炭素数が１～６のアルキル基で置換されていてもよいカルボキシ基、（ｅ’）置換されていてもよい炭素数が１～６のアルキル基で置換されていてもよいカルバモイル基、（ｆ’）置換されていてもよい炭素数が１～６のアルキル基で置換されていてもよいヒドロキシ基、（ｇ’）置換されていてもよい炭素数が１～６のアルキル基で置換されていてもよいメルカプト基、（ｈ’）置換されていてもよい炭素数が１～６のアルキルスルホニル基、（ｉ’）置換されていてもよい炭素数が１～６のアルキル基で置換されていてもよいスルホ基、（ｊ’）ハロゲン原子、（ｋ’）シアノ基、（ｌ’）ニトロ基。 More preferably, in the substituent group A, 1 to 5 substituents selected from the following substituents (c ′) to (l ′) are substituted with the above substituents (A ), (E), (F), (J), (Z), (R), (V), (Z) and (AA);
(C ′) an optionally substituted alkylcarbonyl group having 2 to 7 carbon atoms, (d ′) an optionally substituted carboxy group having 1 to 6 carbon atoms, (E ′) an optionally substituted carbamoyl group having 1 to 6 carbon atoms, (f ′) an optionally substituted alkyl group having 1 to 6 carbon atoms A hydroxy group which may be substituted, (g ′) a mercapto group which may be substituted with an alkyl group having 1 to 6 carbon atoms which may be substituted, and (h ′) a carbon number which may be substituted. 1 to 6 alkylsulfonyl groups, (i ′) an optionally substituted sulfo group having 1 to 6 carbon atoms, (j ′) a halogen atom, (k ′) a cyano group , (L ′) Nitro group.
　置換基Ａ群は、さらに好ましくは、上記置換基（ｃ’）～（ｌ’）から選ばれる１～５個の置換基でアルキル上が置換されていてもよい上記置換基（Ｒ）である。 The substituent group A is more preferably the above substituent (R) which may be substituted on the alkyl with 1 to 5 substituents selected from the above substituents (c ′) to (l ′). .
　上記における「置換されていてもよい炭素数が１～６のアルキル基」、「置換されていてもよい炭素数が２～６のアルケニル基」、「置換されていてもよい炭素数が２～６のアルキニル基」、「置換されていてもよい炭素数が６～１４のアリール基」、「置換されていてもよい炭素数が２～７のアルキルカルボニル基」及び「置換されていてもよい炭素数が１～６のアルキルスルホニル基」の置換基としては、例えば、下記（１）～（１５）からなる置換基Ｂ群が挙げられる；
置換基群Ｂ：
（１）炭素数が１～６のアルキル基、（２）炭素数が２～６のアルケニル基、（３）炭素数が２～６のアルキニル基、（４）炭素数が６～１４のアリール基、（５）ホルミル基、（６）炭素数が２～７のアルキルカルボニル基、（７）炭素数が１～６のアルキル基、炭素数が２～６のアルケニル基、炭素数が２～６のアルキニル基、及び炭素数が６～１４のアリール基から選ばれる置換基で置換されていてもよいカルボキシ基、（８）炭素数が１～６のアルキル基、炭素数が２～６のアルケニル基、炭素数が２～６のアルキニル基、及び炭素数が６～１４のアリール基から選ばれる置換基で置換されていてもよいカルバモイル基、（９）炭素数が１～６のアルキル基、炭素数が２～６のアルケニル基、炭素数が２～６のアルキニル基、及び炭素数が６～１４のアリール基から選ばれる置換基で置換されていてもよいヒドロキシ基、（１０）炭素数が１～６のアルキル基、炭素数が２～６のアルケニル基、炭素数が２～６のアルキニル基、及び炭素数が６～１４のアリール基から選ばれる置換基で置換されていてもよいメルカプト基、（１１）炭素数が１～６のアルキルスルホニル基、（１２）炭素数が１～６のアルキル基で置換されていてもよいスルホ基、（１３）ハロゲン原子、（１４）シアノ基、（１５）ニトロ基等が挙げられる。
　置換基数は特に限定されるものではないが、１～５であることが好ましい。 In the above, “optionally substituted alkyl group having 1 to 6 carbon atoms”, “optionally substituted alkenyl group having 2 to 6 carbon atoms”, “optionally substituted carbon number of 2 to 6” 6 alkynyl group ”,“ optionally substituted aryl group having 6 to 14 carbon atoms ”,“ optionally substituted alkylcarbonyl group having 2 to 7 carbon atoms ”and“ optionally substituted ” Examples of the substituent of the “alkylsulfonyl group having 1 to 6 carbon atoms” include the substituent group B consisting of the following (1) to (15);
Substituent group B:
(1) an alkyl group having 1 to 6 carbon atoms, (2) an alkenyl group having 2 to 6 carbon atoms, (3) an alkynyl group having 2 to 6 carbon atoms, and (4) an aryl having 6 to 14 carbon atoms. Group (5) formyl group, (6) alkylcarbonyl group having 2 to 7 carbon atoms, (7) alkyl group having 1 to 6 carbon atoms, alkenyl group having 2 to 6 carbon atoms, 2 to A carboxy group optionally substituted with a substituent selected from an alkynyl group having 6 carbon atoms and an aryl group having 6 to 14 carbon atoms, (8) an alkyl group having 1 to 6 carbon atoms, and having 2 to 6 carbon atoms An alkamoyl group, a carbamoyl group optionally substituted with a substituent selected from an alkynyl group having 2 to 6 carbon atoms, and an aryl group having 6 to 14 carbon atoms, (9) 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, and carbon A hydroxy group which may be substituted with a substituent selected from an aryl group having 6 to 14 carbon atoms, (10) an alkyl group having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, and 2 carbon atoms A mercapto group optionally substituted with a substituent selected from an alkynyl group having 6 to 6 carbon atoms and an aryl group having 6 to 14 carbon atoms, (11) an alkylsulfonyl group having 1 to 6 carbon atoms, and (12) a carbon number Includes a sulfo group optionally substituted with 1 to 6 alkyl groups, (13) a halogen atom, (14) a cyano group, and (15) a nitro group.
The number of substituents is not particularly limited, but is preferably 1 to 5.
　本明細書における「炭素数が２～６のアルケニル基」とは、少なくとも１個以上の二重結合を有する、炭素数が２～６の直鎖、分枝鎖又は環状の部分不飽和炭化水素基を意味し、例えば、エテニル、１－プロペニル、２－プロペニル、２－メチル－１－プロペニル、１－ブテニル、２－ブテニル、３－ブテニル、３－メチル－２－ブテニル、１－ペンテニル、２－ペンテニル、３－ペンテニル、４－ペンテニル、４－メチル－３－ペンテニル、１－ヘキセニル、３－ヘキセニル、５－ヘキセニル、４－メチル－２－ペンチニル等が挙げられる。 In the present specification, the “alkenyl group having 2 to 6 carbon atoms” refers to a straight, branched or cyclic partially unsaturated hydrocarbon having 2 to 6 carbon atoms having at least one double bond. Means a group such as ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 3-methyl-2-butenyl, 1-pentenyl, 2 -Pentenyl, 3-pentenyl, 4-pentenyl, 4-methyl-3-pentenyl, 1-hexenyl, 3-hexenyl, 5-hexenyl, 4-methyl-2-pentynyl and the like.
　本明細書における「炭素数が２～６のアルキニル基」とは、少なくとも１個以上の三重結合を有する、炭素数が２～６の直鎖、分枝鎖又は環状の部分不飽和炭化水素基を意味し、例えば、エチニル、１－プロピニル、２－プロピニル、１－ブチニル、２－ブチニル、３－ブチニル、１－ペンチニル、２－ペンチニル、３－ペンチニル、４－ペンチニル、１－ヘキシニル、２－ヘキシニル、３－ヘキシニル、４－ヘキシニル、５－ヘキシニル、４－メチル－２－ペンチニル等が挙げられる。 In the present specification, the “alkynyl group having 2 to 6 carbon atoms” means a linear, branched or cyclic partially unsaturated hydrocarbon group having 2 to 6 carbon atoms and having at least one triple bond. For example, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2- Examples include hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl, 4-methyl-2-pentynyl and the like.
　本明細書における「炭素数が２～７のアルキルカルボニル基」とは、上記「炭素数が１～６のアルキル基」がカルボニルを介して結合する基を意味し、例えば、アセチル、プロピオニル、ブチリル、イソブチリル、ｓｅｃ－ブチルカルボニル、ｔｅｒｔ－ブチルカルボニル、ペンタノイル、ヘキサノイル等が挙げられる。 In the present specification, the “alkylcarbonyl group having 2 to 7 carbon atoms” means a group to which the above “alkyl group having 1 to 6 carbon atoms” is bonded via carbonyl, such as acetyl, propionyl, butyryl. , Isobutyryl, sec-butylcarbonyl, tert-butylcarbonyl, pentanoyl, hexanoyl and the like.
　本明細書における「炭素数が１～６のアルキルスルホニル基」とは、上記「炭素数が１～６のアルキル基」がスルホニルを介して結合する基を意味し、例えば、メチルスルホニル、エチルスルホニル、プロピルスルホニル、イソプロピルスルホニル、ブチルスルホニル、イソブチルスルホニル、ｓｅｃ－ブチルスルホニル、ｔｅｒｔ－ブチルスルホニル、ペンチルスルホニル等が挙げられる。 In the present specification, the “alkylsulfonyl group having 1 to 6 carbon atoms” means a group to which the above “alkyl group having 1 to 6 carbon atoms” is bonded via sulfonyl, for example, methylsulfonyl, ethylsulfonyl Propylsulfonyl, isopropylsulfonyl, butylsulfonyl, isobutylsulfonyl, sec-butylsulfonyl, tert-butylsulfonyl, pentylsulfonyl and the like.
　本発明のペプチド化合物又はその塩の分子量は、１００，０００未満であることが好ましく、８０，０００未満であることがより好ましく、７０，０００未満であることがさらに好ましく、６０，０００未満であることがなお一層好ましい。 The molecular weight of the peptide compound of the present invention or a salt thereof is preferably less than 100,000, more preferably less than 80,000, still more preferably less than 70,000, and less than 60,000. Even more preferred.
　ある好ましい実施形態では、本発明のペプチド化合物又はその塩は、式（Ｉａ）又は式（Ｉｂ）で表される化合物である。 In a preferred embodiment, the peptide compound of the present invention or a salt thereof is a compound represented by the formula (Ia) or the formula (Ib).
Figure JPOXMLDOC01-appb-C000023
Figure JPOXMLDOC01-appb-C000023
［式中、各記号は上記の定義の通りである。］ [Wherein each symbol is as defined above. ]
　Ｒ１は、前記と同義であり、それぞれ独立して、ホウ素クラスターからなる１価の基又はそれがリンカー構造を介して結合する１価の基で置換されたα－アミノ酸の側鎖を示す。「ホウ素クラスターからなる１価の基」、「リンカー構造」及び「α－アミノ酸の側鎖」についても前記と同義である。 R 1 has the same meaning as described above, and each independently represents a side chain of an α-amino acid substituted with a monovalent group consisting of a boron cluster or a monovalent group to which it is bonded via a linker structure. The “monovalent group consisting of boron clusters”, “linker structure”, and “side chain of α-amino acid” have the same meaning as described above.
　Ｒ２は、それぞれ独立して、置換基を有していてもよい、中性アミノ酸及び酸性アミノ酸から選ばれるα－アミノ酸の側鎖を示す。 R 2 independently represents a side chain of an α-amino acid selected from a neutral amino acid and an acidic amino acid, which may have a substituent.
　ある実施形態では、Ｒ２の「α－アミノ酸」は、好ましくは、置換されている場合は、セリン、トレオニン、システイン、チロシン、アスパラギン酸、グルタミン酸、ホモセリン、ホモシステイン、ペニシラミン等の側鎖にカルボキシ基、ヒドロキシ基、メルカプト基等の活性基を有するα－アミノ酸であり、無置換の場合は、任意のα－アミノ酸である。 In certain embodiments, the “α-amino acid” of R 2 is preferably substituted with carboxy in the side chain of serine, threonine, cysteine, tyrosine, aspartic acid, glutamic acid, homoserine, homocysteine, penicillamine, etc. An α-amino acid having an active group such as a group, a hydroxy group, a mercapto group, etc., and an unsubstituted α-amino acid when unsubstituted.
　上記の実施形態において、Ｒ２の「α－アミノ酸」は、より好ましくは、置換されている場合は、セリン、トレオニン、システイン、チロシン、アスパラギン酸、グルタミン酸、ホモセリン、ホモシステイン、ペニシラミン等の側鎖にカルボキシ基、ヒドロキシ基、メルカプト基等の活性基を有するα－アミノ酸であり、無置換の場合は、グリシン、アラニン、バリン、ロイシン、イソロイシン、セリン、トレオニン、システイン、メチオニン、アスパラギン、グルタミン、フェニルアラニン、チロシン、アスパラギン酸、グルタミン酸、ホモセリン、ノルロイシン、ノルバリン、チロニン、シトルリン、α-アミノ酪酸、ホモシステイン、ペニシラミン等のα－アミノ酸である。 In the above embodiment, the “α-amino acid” of R 2 is more preferably a side chain such as serine, threonine, cysteine, tyrosine, aspartic acid, glutamic acid, homoserine, homocysteine, penicillamine, etc., when substituted. Is an α-amino acid having an active group such as carboxy group, hydroxy group, mercapto group, etc., and when unsubstituted, glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, asparagine, glutamine, phenylalanine , Tyrosine, aspartic acid, glutamic acid, homoserine, norleucine, norvaline, thyronine, citrulline, α-aminobutyric acid, homocysteine, penicillamine and the like.
　別の実施形態では、Ｒ２の「α－アミノ酸」は、好ましくは、セリン、トレオニン、システイン、チロシン、アスパラギン酸、グルタミン酸、ホモセリン、ホモシステイン、ペニシラミン等の側鎖にカルボキシ基、ヒドロキシ基、メルカプト基等の活性基を有するα－アミノ酸であり；なかでも、グルタミン酸及びアスパラギン酸から選ばれることがより好ましく；グルタミン酸であることが特に好ましい。 In another embodiment, the “α-amino acid” of R 2 is preferably a serine, threonine, cysteine, tyrosine, aspartic acid, glutamic acid, homoserine, homocysteine, penicillamine, etc. in the side chain such as carboxy group, hydroxy group, mercapto An α-amino acid having an active group such as a group; among them, more preferably selected from glutamic acid and aspartic acid; and particularly preferably glutamic acid.
　Ｒ２のα－アミノ酸の側鎖は、それが結合する炭素原子及び該炭素原子に隣接する窒素原子と一緒になって、環を形成していてもよい。ここにおける環状部分としては、例えば、プロリンの環状部分（即ち、ピロリジン環）が挙げられる。ある実施形態では、Ｒ２のα－アミノ酸の側鎖は、それが結合する炭素原子及び該炭素原子に隣接する窒素原子と一緒になって環を形成しない。 The side chain of the α-amino acid of R 2 may be combined with the carbon atom to which it is attached and the nitrogen atom adjacent to the carbon atom to form a ring. Examples of the cyclic moiety herein include a proline cyclic moiety (that is, a pyrrolidine ring). In certain embodiments, the side chain of the α-amino acid of R 2 does not form a ring with the carbon atom to which it is attached and the nitrogen atom adjacent to the carbon atom.
　Ｒ２のα－アミノ酸の側鎖の置換基としては、ポリエーテルからなる１価の基に加えて、上記その他の置換基と同様のものが挙げられる。ある実施形態では、Ｒ２において置換基は存在しない。 Examples of the substituent on the side chain of the α-amino acid of R 2 include those similar to the above-mentioned other substituents in addition to the monovalent group consisting of polyether. In one embodiment, the substituent at R 2 is absent.
　Ｒ２においてα－アミノ酸の側鎖は、置換基により、例えば、炭素原子、窒素原子又は硫黄原子上の任意のＨ又はＯＨが置換されていてもよく、好ましくは、アミノ酸の活性基のＨ及びＯＨ（例えば、カルボキシ基のＯＨ並びにヒドロキシ基及びメルカプト基のＨ）から選ばれる基が置換されていてもよく、より好ましくは、カルボキシ基のＯＨが置換されていてもよい。 In R 2 , the side chain of the α-amino acid may be substituted with any H or OH on the carbon atom, nitrogen atom or sulfur atom, for example, preferably H and OH of the amino acid active group. A group selected from OH (for example, OH of a carboxy group and H of a hydroxy group and a mercapto group) may be substituted, and more preferably, OH of a carboxy group may be substituted.
　Ｒ２において置換基が、カルボキシ基、ヒドロキシ基及びメルカプト基との間で形成し得る結合としては、それぞれ、ホウ素クラスター（又はリンカー構造）が形成し得る結合として上記で挙げたものと同様のものが挙げられる。 As the bond that the substituent in R 2 can form with a carboxy group, a hydroxy group, and a mercapto group, the same bonds as those described above as the bonds that can be formed with a boron cluster (or linker structure), respectively. Is mentioned.
　Ｒ２における「ポリエーテルからなる１価の基」の具体例としては、上記式（Ｐ１）～（Ｐ３）の何れかで表される基が挙げられる。中でも好ましくは、上記式（Ｐ１）で表される基である。 Specific examples of the “monovalent group consisting of polyether” in R 2 include groups represented by any of the above formulas (P1) to (P3). Among them, a group represented by the above formula (P1) is preferable.
　ｂが１以上の場合、Ｒ１を有するα－アミノ酸残基とＲ２を有するα－アミノ酸残基とは、任意に共重合しており、ランダム共重合していても、ブロック共重合していても、交互共重合していてもよい。 When b is 1 or more, the α-amino acid residue having R 1 and the α-amino acid residue having R 2 are arbitrarily copolymerized, and even if they are randomly copolymerized, they are block-copolymerized. Alternatively, alternating copolymerization may be performed.
　ある実施形態では、Ｒ１の「α－アミノ酸」及びＲ２の「α－アミノ酸」の全てが同一のα－アミノ酸である。 In some embodiments, all of the “α-amino acid” of R 1 and the “α-amino acid” of R 2 are the same α-amino acid.
　Ｘ１、及びｃが１の場合のＹ２は、それぞれ独立して、（１）水素原子、（２）炭素数が１～６のアルキル基、（３）炭素数が２～７のアルキルカルボニル基、又は（４）ポリエーテルからなる１価の基を示す。 X 1 and Y 2 when c is 1 are each independently (1) a hydrogen atom, (2) an alkyl group having 1 to 6 carbon atoms, and (3) an alkylcarbonyl having 2 to 7 carbon atoms. A monovalent group consisting of a group or (4) a polyether;
　Ｘ１は、好ましくは、水素原子、又はポリエーテルからなる１価の基であり、さらに好ましくは、水素原子である。ｃが１の場合のＹ２は、好ましくは、水素原子、又はポリエーテルからなる１価の基であり、さらに好ましくは、ポリエーテルからなる１価の基である。 X 1 is preferably a hydrogen atom or a monovalent group consisting of a polyether, and more preferably a hydrogen atom. Y 2 in the case where c is 1 is preferably a hydrogen atom or a monovalent group consisting of a polyether, and more preferably a monovalent group consisting of a polyether.
　Ｘ２、及びｃが１の場合のＹ１は、それぞれ独立して、（１）炭素数が１～６のアルキル基で置換されていてもよいヒドロキシ基、（２）炭素数が１～６のアルキル基で置換されていてもよいアミノ基、又は（３）ポリエーテルからなる１価の基を示す。 X 2 and Y 1 when c is 1 are each independently (1) a hydroxy group which may be substituted with an alkyl group having 1 to 6 carbon atoms, and (2) a carbon number of 1 to 6 The monovalent group which consists of the amino group which may be substituted by the alkyl group of (3), or polyether.
　Ｘ２は、好ましくは、炭素数が１～６のアルキル基で置換されていてもよいヒドロキシ基、又は炭素数が１～６のアルキル基で置換されていてもよいアミノ基である。ｃが１の場合のＹ１は、好ましくは、ポリエーテルからなる１価の基である。 X 2 is preferably a hydroxy group which may be substituted with an alkyl group having 1 to 6 carbon atoms, or an amino group which may be substituted with an alkyl group having 1 to 6 carbon atoms. When c is 1, Y 1 is preferably a monovalent group made of a polyether.
　ｃが２以上の場合のＹ１及びＹ２は、それぞれ、ポリエーテルからなるｃ価の基を示す。 When c is 2 or more, Y 1 and Y 2 each represent a c-valent group made of a polyether.
　Ｘ１、Ｘ２、Ｙ１又はＹ２がポリエーテルからなる１価の基又はポリエーテルからなるｃ価の基の場合、それらが、アミノ基（アミノ酸残基のＮＨ）及びカルボキシ基（アミノ酸残基のＣＯ）との間で形成し得る結合としては、それぞれ、ホウ素クラスター（又はリンカー構造）が形成し得る結合として上記で挙げたものと同様のものが挙げられる。 When X 1 , X 2 , Y 1 or Y 2 is a monovalent group consisting of a polyether or a c-valent group consisting of a polyether, these are an amino group (NH of an amino acid residue) and a carboxy group (amino acid residue). Examples of the bond that can be formed with the group (CO) include the same bonds as those described above as the bonds that the boron cluster (or linker structure) can form.
　Ｘ１、Ｘ２、Ｙ１及びＹ２における「ポリエーテルからなる１価の基」の具体例としては、上記式（Ｐ１）～（Ｐ３）の何れかで表される基が挙げられる。中でも好ましくは、上記式（Ｐ１）で表される基である。Ｙ１及びＹ２における「ポリエーテルからなるｃ価の基」の具体例としては、ｃが２の場合、上記式（Ｐ４）又は（Ｐ５）で表される基が挙げられる。 Specific examples of the “monovalent group consisting of polyether” in X 1 , X 2 , Y 1 and Y 2 include groups represented by any of the above formulas (P1) to (P3). Among them, a group represented by the above formula (P1) is preferable. Specific examples of the “c-valent group made of polyether” in Y 1 and Y 2 include a group represented by the above formula (P4) or (P5) when c is 2.
　式（Ｉａ）においては、Ｒ２のα－アミノ酸の置換基、Ｘ１及びＹ１のうち少なくともいずれか１つが、式（Ｉｂ）においては、Ｒ２のα－アミノ酸の置換基、Ｘ２及びＹ２のうち少なくともいずれか１つが、ポリエーテルからなる１価の基又はポリエーテルからなるｃ価の基である。 In the formula (Ia), at least one of the substituents of the α-amino acid of R 2 , X 1 and Y 1 , and in the formula (Ib), the substituent of the α-amino acid of R 2 , X 2 and At least one of Y 2 is a monovalent group made of a polyether or a c-valent group made of a polyether.
　ａとｂの合計は、２以上の整数であり、好ましくは、３以上の整数であり、より好ましくは、５以上の整数であり、さらに好ましくは、１０以上の整数である。ａとｂの合計は、好ましくは、２００以下の整数であり、より好ましくは、１５０以下の整数であり、さらに好ましくは、１００以下の整数である。 The total of a and b is an integer of 2 or more, preferably an integer of 3 or more, more preferably an integer of 5 or more, and further preferably an integer of 10 or more. The sum of a and b is preferably an integer of 200 or less, more preferably an integer of 150 or less, and still more preferably an integer of 100 or less.
　ａとｂの合計に対するａの比は、好ましくは、０．１以上であり、より好ましくは、０．２以上であり、さらに好ましくは、０．３以上である。 The ratio of a to the sum of a and b is preferably 0.1 or more, more preferably 0.2 or more, and still more preferably 0.3 or more.
　ｃは、１以上の整数であり、好ましくは、１～１０の整数であり、より好ましくは、１～５の整数であり、さらに好ましくは、１又は２であり、特に好ましくは、１である（即ち、式（Ｉａ）及び（Ｉｂ）が、それぞれ、式（ＩＡ）及び式（ＩＢ）： c is an integer of 1 or more, preferably an integer of 1 to 10, more preferably an integer of 1 to 5, still more preferably 1 or 2, and particularly preferably 1. (Ie, formulas (Ia) and (Ib) are represented by formulas (IA) and (IB), respectively:
Figure JPOXMLDOC01-appb-C000024
Figure JPOXMLDOC01-appb-C000024
［式中、各記号は上記定義の通りである。］
で表される。）。 [Wherein the symbols are as defined above. ]
It is represented by ).
　本発明のペプチド化合物は塩であってもよい。塩としては、例えば、金属塩、アンモニウム塩、有機塩基との塩、無機酸との塩、有機酸との塩、塩基性アミノ酸又は酸性アミノ酸との塩等が挙げられる。金属塩の好適な例としては、例えば、ナトリウム塩、カリウム塩等のアルカリ金属塩；カルシウム塩、マグネシウム塩等のアルカリ土類金属塩；アルミニウム塩等が挙げられる。アンモニウム塩の好適な例としては、例えば、テトラメチルアンモニウム塩、テトラブチルアンモニウム塩、ポリアミン（例えば、１，４－ジアミノブタン、スペルミン、スペルミジン）の１価又は多価のアンモニウム塩等が挙げられる。有機塩基との塩の好適な例としては、例えば、トリメチルアミン、トリエチルアミン、ピリジン、ピコリン、２，６－ルチジン、エタノールアミン、ジエタノールアミン、トリエタノールアミン、シクロヘキシルアミン、ジシクロヘキシルアミン、Ｎ，Ｎ’－ジベンジルエチレンジアミン、ポリアミン（例えば、１，４－ジアミノブタン、スペルミン、スペルミジン）等のアミン類との塩が挙げられる。無機酸との塩の好適な例としては、例えば、塩酸、臭化水素酸、硝酸、硫酸、リン酸等との塩が挙げられる。有機酸との塩の好適な例としては、例えば、ギ酸、酢酸、トリフルオロ酢酸、フタル酸、フマル酸、シュウ酸、酒石酸、マレイン酸、クエン酸、コハク酸、リンゴ酸、メタンスルホン酸、ベンゼンスルホン酸、ｐ－トルエンスルホン酸等との塩が挙げられる。塩基性アミノ酸との塩の好適な例としては、例えば、アルギニン、リジン、オルニチン等との塩が挙げられ、酸性アミノ酸との塩の好適な例としては、例えば、アスパラギン酸、グルタミン酸等との塩が挙げられる。本発明のペプチド化合物の塩は、中でも、好ましくは、薬学的に許容される塩であることが好ましい。 The peptide compound of the present invention may be a salt. Examples of the salt include metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic amino acids or acidic amino acids, and the like. Preferable examples of the metal salt include alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt and magnesium salt; aluminum salt and the like. Preferable examples of the ammonium salt include tetramethylammonium salt, tetrabutylammonium salt, and monovalent or polyvalent ammonium salt of polyamine (eg, 1,4-diaminobutane, spermine, spermidine). Preferable examples of the salt with an organic base include, for example, trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N, N′-dibenzyl. Examples thereof include salts with amines such as ethylenediamine and polyamine (eg, 1,4-diaminobutane, spermine, spermidine). Preferable examples of the salt with inorganic acid include salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid and the like. Preferable examples of the salt with organic acid include, for example, formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzene And salts with sulfonic acid, p-toluenesulfonic acid and the like. Preferable examples of salts with basic amino acids include salts with arginine, lysine, ornithine and the like, and preferable examples of salts with acidic amino acids include salts with aspartic acid, glutamic acid and the like. Is mentioned. Among them, the salt of the peptide compound of the present invention is preferably a pharmaceutically acceptable salt.
　本発明のペプチド化合物又はその塩（以下、本発明の化合物という）は、腫瘍疾患のホウ素中性子捕捉療法に用いることができる。 The peptide compound of the present invention or a salt thereof (hereinafter referred to as the compound of the present invention) can be used for boron neutron capture therapy for tumor diseases.
　本明細書における「腫瘍疾患」としては、悪性黒色腫（メラノーマ）、腎癌、前立腺癌、乳癌、肺癌、膵癌、大腸癌、肝細胞癌、胆道癌、胃癌、卵巣癌、食道癌、尿路上皮癌、結腸癌、骨癌、皮膚癌（例えば悪性皮膚癌）、頭頚部癌、子宮癌、直腸癌、肛門部癌、精巣癌、子宮癌、卵管癌、子宮内膜癌、子宮頚部癌、膣癌、小腸癌、内分泌系癌、甲状腺癌、副甲状腺癌、副腎癌、柔組織肉腫、尿道癌、陰茎癌、小児固形癌、膀胱癌、悪性胸膜中皮腫、脳腫瘍（例えば悪性脳腫瘍）、中枢神経系腫瘍等が挙げられる。 As used herein, "tumor disease" includes malignant melanoma, renal cancer, prostate cancer, breast cancer, lung cancer, pancreatic cancer, colon cancer, hepatocellular carcinoma, biliary tract cancer, stomach cancer, ovarian cancer, esophageal cancer, urinary tract Skin cancer, colon cancer, bone cancer, skin cancer (eg malignant skin cancer), head and neck cancer, uterine cancer, rectal cancer, anal cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer , Vaginal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, childhood solid cancer, bladder cancer, malignant pleural mesothelioma, brain tumor (eg malignant brain tumor) And central nervous system tumors.
　本発明の化合物を用いるホウ素中性子捕捉療法は、例えば、腫瘍疾患を患う哺乳動物（例えばヒト）に、化合物が標的部位に蓄積するような任意の適当な投与経路により本発明の化合物を含む薬剤を投与することによって行われる。本発明の化合物は、腫瘍に選択的に蓄積されていることが好ましい。化合物を含む製剤は一度に投与してもよいし、或いは順次投与してもよい。製剤の投与を必要に応じて繰り返してもよい。 Boron neutron capture therapy using a compound of the present invention can be performed, for example, on a mammal (eg, a human) suffering from a tumor disease by applying a drug containing the compound of the present invention by any suitable route of administration such that the compound accumulates at the target site. By administration. The compound of the present invention is preferably accumulated selectively in a tumor. Formulations containing the compound may be administered at once or sequentially. Administration of the formulation may be repeated as necessary.
　腫瘍内に本発明の化合物が到達した後、その部位に有効量の、熱中性子線或いは熱外中性子線のような低エネルギー中性子線を照射する。皮膚を通してその部位を照射してもよいし、或いはその部位を照射前に完全に或いは部分的に暴露して照射してもよい。また、同時に複数の方向から照射してもよい。本発明の化合物の投与とそれに続く熱中性子線或いは熱外中性子線の照射を必要に応じて繰り返してもよい。例えば数か月程度の間隔で、複数回照射してもよい。照射回数の総数は、好ましくは、１～１０回、より好ましくは１～５回である。 After the compound of the present invention has reached the tumor, the site is irradiated with an effective amount of a low energy neutron beam such as a thermal neutron beam or an epithermal neutron beam. The site may be irradiated through the skin, or the site may be irradiated completely or partially exposed prior to irradiation. Moreover, you may irradiate from several directions simultaneously. Administration of the compound of the present invention and subsequent irradiation with thermal neutron rays or epithermal neutron rays may be repeated as necessary. For example, you may irradiate several times at intervals of about several months. The total number of times of irradiation is preferably 1 to 10 times, more preferably 1 to 5 times.
　本発明の化合物の投与経路は、好ましくは、静脈内投与、動脈内投与、筋肉内投与、皮下投与、皮内投与、脊髄内投与、腹腔内投与である。 The administration route of the compound of the present invention is preferably intravenous administration, intraarterial administration, intramuscular administration, subcutaneous administration, intradermal administration, intraspinal administration, intraperitoneal administration.
　本発明の化合物は、熱中性子線或いは熱外中性子線の照射前の７２時間以内（好ましくは、５分～４８時間前、より好ましくは、３０分～３０時間前）に、腫瘍疾患を患う哺乳動物に投与され得る。また、必要に応じて、熱中性子線或いは熱外中性子線の照射中に投与してもよい。本発明の化合物の代表的な用量は、好ましくは、１回の熱中性子線或いは熱外中性子線の照射につき０．０１　ｍｇ～５０　ｇ／体重ｋｇの範囲内である。ホウ素クラスター換算では、好ましくは、１回の熱中性子線或いは熱外中性子線の照射につき０．００５　ｍｇ～２５　ｇ／体重ｋｇの範囲内である。１回の熱中性子線或いは熱外中性子線の照射時間は、好ましくは、１分～５時間、より好ましくは、１０分～２時間である。熱中性子線或いは熱外中性子線の照射量は、特に限定されるものではなく、中性子補捉療法で用いられる一般的な照射量であればよい。 The compound of the present invention is a mammal suffering from a tumor disease within 72 hours (preferably 5 minutes to 48 hours, more preferably 30 minutes to 30 hours before) of irradiation with thermal neutrons or epithermal neutrons. Can be administered to animals. Moreover, you may administer during the irradiation of a thermal neutron beam or an epithermal neutron beam as needed. A typical dose of the compound of the present invention is preferably in the range of 0.01 mg to 50 g / kg of body weight per irradiation of thermal neutron beam or epithermal neutron beam. In terms of boron cluster, it is preferably in the range of 0.005 mg to 25 g / kg of body weight per irradiation of thermal neutron beam or epithermal neutron beam. The irradiation time of one thermal neutron beam or epithermal neutron beam is preferably 1 minute to 5 hours, more preferably 10 minutes to 2 hours. The irradiation amount of the thermal neutron beam or the epithermal neutron beam is not particularly limited as long as it is a general irradiation amount used in neutron capture therapy.
　本発明の化合物を用いるホウ素中性子捕捉療法は、単独の治療として適用してもよいし、或いは従来の手術又は化学療法と共に適用してもよい。所望であれば、腫瘍を外科的に可能な程度に除去した後、本発明の化合物を用いるホウ素中性子捕捉療法により残りの腫瘍を破壊することも可能である。 Boron neutron capture therapy using the compounds of the present invention may be applied as a single treatment or may be applied with conventional surgery or chemotherapy. If desired, after removing the tumor to the extent possible surgically, the remaining tumor can be destroyed by boron neutron capture therapy using the compounds of the present invention.
　ホウ素中性子捕捉療法において、本発明の化合物は、ＢＰＡ（ｐ－ボロノフェニルアラニン）やＢＳＨ（メルカプトウンデカハイドロドデカボレート）のような他のホウ素化合物（以下、併用薬物という）と共に用いることもできる。併用薬物を用いる場合、本発明の化合物と併用薬物の投与時期は限定されず、本発明の化合物及び併用薬物を、腫瘍疾患を患う哺乳動物に対し、同時に投与してもよいし、時間差をおいて投与してもよい。時間差をおいて投与する場合、本発明の化合物、併用薬物の順で投与してもよいし、或いはその逆の順で投与してもよい。 In boron neutron capture therapy, the compound of the present invention can be used together with other boron compounds (hereinafter referred to as concomitant drugs) such as BPA (p-boronophenylalanine) and BSH (mercaptoundecahydrododecaborate). When a concomitant drug is used, the administration time of the compound of the present invention and the concomitant drug is not limited, and the compound of the present invention and the concomitant drug may be administered simultaneously to a mammal suffering from a tumor disease, with a time difference. May be administered. When administering at a time difference, the compound of the present invention and the concomitant drug may be administered in this order, or vice versa.
　本発明の化合物は、薬学的に許容される溶媒、賦形剤、結合剤、安定化剤、分散剤等の薬学的に許容される担体とともに、注射用溶液、懸濁液、乳剤、クリーム剤、軟膏剤、吸入剤、坐剤等の非経口剤形に製剤化してもよい。本発明の化合物は、注射用溶液に製剤化することが好ましい。 The compound of the present invention contains a pharmaceutically acceptable carrier, excipient, binder, stabilizer, dispersant and the like, together with a pharmaceutically acceptable carrier, an injectable solution, suspension, emulsion, cream. And may be formulated into parenteral dosage forms such as ointments, inhalants, suppositories, and the like. The compound of the present invention is preferably formulated into an injectable solution.
　注射用溶液は、薬学的に受容可能な溶媒中で、本発明の化合物の溶液として製造され得る。それらの溶液はまた、安定化成分及び／又は緩衝化成分を含んでいてもよい。また、注射用溶液は、使用前に適切な溶媒を加えて用いる乾燥製剤であってもよい。 Injection solutions can be prepared as solutions of the compounds of the invention in pharmaceutically acceptable solvents. These solutions may also contain stabilizing components and / or buffering components. In addition, the injection solution may be a dry preparation to which an appropriate solvent is added before use.
　次に、本発明の化合物の製造法について説明する。 Next, a method for producing the compound of the present invention will be described.
　本発明のペプチド化合物の中でも、式（Ｉａ’）及び（Ｉｂ’）で表される化合物（以下、化合物（Ｉａ’）、化合物（Ｉｂ’）という）は、式（ＩＩａ）及び（ＩＩｂ）で表される化合物（以下、化合物（ＩＩａ）又は化合物（ＩＩｂ）という）のα－アミノ酸の側鎖と、式（Ｂ）で表される化合物（以下、化合物（Ｂ）という）とを、Ａ２及びＡ１の部分で、公知の縮合反応、付加反応及び置換反応によりカップリングさせてＡを形成し、それにより、直接或いはリンカー構造を介して結合させることにより得ることができる。その際、反応基質にカルボキシ基、ヒドロキシ基、アミノ基、メルカプト基のような活性基が存在する場合は、これらの官能基を、予め公知の保護基で保護しておいてもよい。 Among the peptide compounds of the present invention, compounds represented by formulas (Ia ′) and (Ib ′) (hereinafter referred to as compounds (Ia ′) and (Ib ′)) are represented by formulas (IIa) and (IIb). The α-amino acid side chain of the compound represented (hereinafter referred to as Compound (IIa) or Compound (IIb)) and the compound represented by Formula (B) (hereinafter referred to as Compound (B)) are converted into A 2. And A 1 can be coupled by a known condensation reaction, addition reaction and substitution reaction to form A, and can be obtained by bonding directly or via a linker structure. At that time, when an active group such as a carboxy group, a hydroxy group, an amino group, or a mercapto group is present on the reaction substrate, these functional groups may be protected in advance with a known protecting group.
Figure JPOXMLDOC01-appb-C000025
Figure JPOXMLDOC01-appb-C000025
［式中、波線部分は、結合手であるか、或いはリンカー構造又はその断片構造を示し、Ａ１は、α－アミノ酸の側鎖、それに結合するリンカー構造又はその断片構造における官能基であり、Ａ２は、ホウ素クラスター、それに結合するリンカー構造又はその断片構造における官能基である。］ [Wherein, the wavy line portion is a bond or represents a linker structure or a fragment structure thereof, and A 1 is a side chain of an α-amino acid, a linker group or a functional group in the fragment structure bound thereto, A 2 is a functional group in a boron cluster, a linker structure bonded to the boron cluster, or a fragment structure thereof. ]
　リンカー構造が、例えば式（Ｌ’）で表される基である場合は、Ａは、Ｐ２とα－アミノ酸の側鎖の一部、或いはＱ２に相当する構造である。例えば、Ａとしては、－Ｏ－、－Ｓ－、－ＮＨ－、－ＳＯ２－、－ＣＯ－、－ＮＨＣＯ－、－ＣＯＮＨ－、－ＯＣＯ－、－ＣＯＯ－、－ＳＣＯ－、－ＣＯＳ－、－ＮＨＣＯＮＨ－、－ＮＨＣＳＮＨ－、－ＯＣＯＮＨ－、－ＮＨＣＯＯ－、－ＳＳ－、－Ｐｈ－、－ＯＰｈＯ－、－ＯＰｈ－、－ＰｈＯ－、－ＳＰｈＳ－、－ＳＰｈ－、－ＰｈＳ－、－ＮＨＰｈＮＨ－、－ＮＨＰｈ－、－ＰｈＮＨ－、－複素環－、－Ｏ－複素環－Ｏ－、－Ｏ－複素環－、－複素環－Ｏ－、－Ｓ－複素環－Ｓ－、－Ｓ－複素環－、－複素環－Ｓ－、－ＮＨ－複素環－ＮＨ－、－ＮＨ－複素環－、－複素環－ＮＨ－等（ここで、Ｐｈは、１，２－フェニレン、１，３－フェニレン又は１，４－フェニレンを示す。）が挙げられる。以下にＡを形成するための方法の一例を挙げる。 When the linker structure is, for example, a group represented by the formula (L ′), A is a part of the side chain of P 2 and α-amino acid, or a structure corresponding to Q 2 . For example, A includes —O—, —S—, —NH—, —SO 2 —, —CO—, —NHCO—, —CONH—, —OCO—, —COO—, —SCO—, —COS—. , —NHCONH—, —NHCSNH—, —OCONH—, —NHCOO—, —SS—, —Ph—, —OPhO—, —OPh—, —PhO—, —SPhS—, —SPh—, —PhS—, — NHPhNH-, -NHPh-, -PhNH-, -heterocyclic-, -O-heterocyclic-O-, -O-heterocyclic-, -heterocyclic-O-, -S-heterocyclic-S-, -S -Heterocycle-, -heterocycle-S-, -NH-heterocycle-NH-, -NH-heterocycle-, -heterocycle-NH- and the like (where Ph is 1,2-phenylene, 1, 3-phenylene or 1,4-phenylene). An example of a method for forming A is given below.
　Ａが、－ＳＳ－の場合は、例えば、Ａ１及びＡ２の一方が、メルカプト基であり、他方が、ピリジン－２－イルジスルファニル基であればよい。化合物（ＩＩａ）又は化合物（ＩＩｂ）と、化合物（Ｂ）を溶媒（例えば、メタノール、エタノールのようなアルコール類、ジメチルスルホキシド（ＤＭＳＯ）のようなスルホキシド類、又はそれらの混合物）中で反応させることにより、化合物（Ｉａ’）又は化合物（Ｉｂ’）を得ることができる。反応温度は、通常－８０から１５０℃である。 When A is —SS—, for example, one of A 1 and A 2 may be a mercapto group and the other may be a pyridin-2-yldisulfanyl group. Reacting compound (IIa) or compound (IIb) with compound (B) in a solvent (for example, alcohols such as methanol and ethanol, sulfoxides such as dimethylsulfoxide (DMSO), or a mixture thereof). Thus, compound (Ia ′) or compound (Ib ′) can be obtained. The reaction temperature is usually from -80 to 150 ° C.
　Ａが、－ＮＨＣＯ－及び－ＣＯＮＨ－の場合は、例えば、Ａ１及びＡ２の一方が、カルボキシ基又はその活性基（例えば、４-ニトロフェノールエステル、ペンタフルオロフェノールエステルのような活性エステルや、酸クロリドのような酸ハライド等）であり、他方が、アミノ基であればよい。化合物（ＩＩａ）又は化合物（ＩＩｂ）と、化合物（Ｂ）を、溶媒（例えば、ジメチルスルホキシド（ＤＭＳＯ）のようなスルホキシド類、Ｎ－メチル－２－ピロリドン（ＮＭＰ）、Ｎ，Ｎ－ジメチルホルムアミド（ＤＭＦ）のようなアミド類、又はそれらの混合物）中、必要に応じて、縮合剤（例えば、１－エチル－３－（３－ジメチルアミノプロピル）カルボジイミド又はその塩のようなカルボジイミド系縮合剤；４－（４，６－ジメトキシ－１，３，５－トリアジン－２－イル）－４－メチルモルホリニウムクロリドのようなトリアジン系縮合剤等）、活性剤（例えば、Ｎ－ヒドロキシスクシンイミド、１－ヒドロキシベンゾトリアゾール（ＨＯＢｔ）、４－ジメチルアミノピリジン（ＤＭＡＰ）等）、塩基（例えば、Ｎ－メチルモルホリン（ＮＭＭ）、トリエチルアミンのような有機塩基等）の存在下で反応させることにより、化合物（Ｉａ’）又は化合物（Ｉｂ’）を得ることができる。反応温度は、通常－８０から１５０℃である。 When A is —NHCO— or —CONH—, for example, one of A 1 and A 2 is a carboxy group or an active group thereof (for example, an active ester such as 4-nitrophenol ester, pentafluorophenol ester, , Acid halide such as acid chloride, etc.) and the other may be an amino group. Compound (IIa) or Compound (IIb) and Compound (B) are mixed with a solvent (for example, sulfoxides such as dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), N, N-dimethylformamide ( Amides such as DMF), or a mixture thereof, if necessary, a condensing agent (for example, a carbodiimide-based condensing agent such as 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide or a salt thereof); 4- (4,6-dimethoxy-1,3,5-triazin-2-yl) -4-methylmorpholinium chloride and the like), an activator (for example, N-hydroxysuccinimide, 1 -Hydroxybenzotriazole (HOBt), 4-dimethylaminopyridine (DMAP), etc.), base (for example, N-methylmol Phosphorus (NMM), by reacting in the presence of an organic base, etc.), such as triethylamine, to give compound (Ia ') or compound (Ib'). The reaction temperature is usually from -80 to 150 ° C.
　Ａが、－ＳＣＯ－及び－ＣＯＳ－の場合は、例えば、Ａ１及びＡ２の一方が、カルボキシ基又はその活性基地（例えば、４-ニトロフェノールエステル、ペンタフルオロフェノールエステルのような活性エステルや、酸クロリドのような酸ハライド等）であり、他方が、メルカプト基であればよい。化合物（ＩＩａ）又は化合物（ＩＩｂ）と、化合物（Ｂ）を、溶媒（例えば、ジメチルスルホキシド（ＤＭＳＯ）のようなスルホキシド類、Ｎ－メチル－２－ピロリドン（ＮＭＰ）、Ｎ，Ｎ－ジメチルホルムアミド（ＤＭＦ）のようなアミド類、又はそれらの混合物）中、必要に応じて、縮合剤（例えば、１－エチル－３－（３－ジメチルアミノプロピル）カルボジイミド又はその塩のようなカルボジイミド系縮合剤；４－（４，６－ジメトキシ－１，３，５－トリアジン－２－イル）－４－メチルモルホリニウムクロリドのようなトリアジン系縮合剤等）、活性剤（例えば、Ｎ－ヒドロキシスクシンイミド、１－ヒドロキシベンゾトリアゾール（ＨＯＢｔ）、４－ジメチルアミノピリジン（ＤＭＡＰ）等）、塩基（例えば、Ｎ－メチルモルホリン（ＮＭＭ）、トリエチルアミンのような有機塩基等）の存在下で反応させることにより、化合物（Ｉａ’）又は化合物（Ｉｂ’）を得ることができる。反応温度は、通常－８０から１５０℃である。 When A is —SCO— or —COS—, for example, one of A 1 and A 2 is a carboxy group or an active base thereof (for example, an active ester such as 4-nitrophenol ester, pentafluorophenol ester, , Acid halide such as acid chloride, etc.) and the other may be a mercapto group. Compound (IIa) or Compound (IIb) and Compound (B) are mixed with a solvent (for example, sulfoxides such as dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), N, N-dimethylformamide ( Amides such as DMF), or a mixture thereof, if necessary, a condensing agent (for example, a carbodiimide-based condensing agent such as 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide or a salt thereof); 4- (4,6-dimethoxy-1,3,5-triazin-2-yl) -4-methylmorpholinium chloride and the like), an activator (for example, N-hydroxysuccinimide, 1 -Hydroxybenzotriazole (HOBt), 4-dimethylaminopyridine (DMAP), etc.), base (for example, N-methylmol Phosphorus (NMM), by reacting in the presence of an organic base, etc.), such as triethylamine, to give compound (Ia ') or compound (Ib'). The reaction temperature is usually from -80 to 150 ° C.
　Ａが、－複素環（１，２，３－トリアゾール）－の場合は、例えば、Ａ１及びＡ２の一方が、アジド（Ｎ３）基であり、他方が、エチニル（ＨＣ≡Ｃ）基であればよい。化合物（ＩＩａ）又は化合物（ＩＩｂ）と、化合物（Ｂ）を、溶媒（例えば、Ｎ，Ｎ－ジメチルホルムアミド（ＤＭＦ）のようなアミド類；アセトンのようなケトン類；メタノール、エタノール、ｔｅｒｔ－ブタノールのようなアルコール類；水（緩衝液を含む）；又はそれらの混合物）中、銅触媒（例えば、硫酸銅（Ｉ）、臭化銅（Ｉ）、ヨウ化銅（Ｉ）、トリフルオロメタンスルホン酸銅（Ｉ）等）、必要に応じて、添加剤（例えば、アスコルビン酸ナトリウム、トリス(２－カルボキシエチル)ホスフィン（ＴＣＥＰ）等の還元剤等）、配位子（例えば、トリス（ベンジルトリアゾリルメチル）アミン（ＴＢＴＡ）、トリス(ヒドロキシプロピルトリアゾリルメチル)アミン（ＴＨＰＴＡ）、バソフェナントロリンジスルホネート（ＢＰＤＳ）等）、塩基（例えば、Ｎ，Ｎ，Ｎ’，Ｎ’’，Ｎ’’－ペンタメチルジエチレントリアミン（ＰＭＤＥＴＡ）、トリエチルアミンのような有機塩基等）の存在下で反応させることにより、化合物（Ｉａ’）又は化合物（Ｉｂ’）を得ることができる。反応温度は、通常－８０から１５０℃である。 When A is -heterocyclic (1,2,3-triazole)-, for example, one of A 1 and A 2 is an azide (N 3 ) group and the other is an ethynyl (HC≡C) group If it is. Compound (IIa) or Compound (IIb) and Compound (B) are mixed with a solvent (for example, amides such as N, N-dimethylformamide (DMF); ketones such as acetone; methanol, ethanol, tert-butanol In water (including buffer); or mixtures thereof) in copper catalysts (eg, copper (I) sulfate, copper (I) bromide, copper (I) iodide, trifluoromethanesulfonic acid) Copper (I) etc.), if necessary, additives (for example, reducing agents such as sodium ascorbate, tris (2-carboxyethyl) phosphine (TCEP), etc.), ligands (for example, tris (benzyltriazoli) Rumethyl) amine (TBTA), tris (hydroxypropyltriazolylmethyl) amine (THPTA), bathophenanthroline disulfonate (BPDS) Etc.), a base (for example, an organic base such as N, N, N ′, N ″, N ″ -pentamethyldiethylenetriamine (PMDETA), triethylamine, etc.) to give a compound (Ia ') Or compound (Ib') can be obtained. The reaction temperature is usually from -80 to 150 ° C.
　また、Ａ１がリンカー構造又はその断片構造に結合している場合、当該リンカー構造又はその断片構造は、上記のＡの構築方法と同様に、或いは公知の方法でα－アミノ酸の側鎖に導入することができる。その際、反応基質にカルボキシ基、ヒドロキシ基、アミノ基、メルカプト基のような活性基が存在する場合は、これらの官能基を、予め公知の保護基で保護しておいてもよい。 Further, when A 1 is bound to a linker structure or a fragment structure thereof, the linker structure or the fragment structure thereof is introduced into the side chain of an α-amino acid in the same manner as in the construction method of A described above or by a known method. can do. At that time, when an active group such as a carboxy group, a hydroxy group, an amino group, or a mercapto group is present on the reaction substrate, these functional groups may be protected in advance with a known protecting group.
　また、Ａ２がリンカー構造又はその断片構造に結合している場合、当該リンカー構造又はその断片構造は、上記のＡの構築方法と同様に、或いは公知の方法でホウ素クラスターに導入することができる。その際、反応基質にカルボキシ基、ヒドロキシ基、アミノ基、メルカプト基のような活性基が存在する場合は、これらの官能基を、予め公知の保護基で保護しておいてもよい。 In addition, when A 2 is bonded to a linker structure or a fragment structure thereof, the linker structure or the fragment structure thereof can be introduced into the boron cluster in the same manner as in the construction method of A described above or by a known method. . At that time, when an active group such as a carboxy group, a hydroxy group, an amino group, or a mercapto group is present on the reaction substrate, these functional groups may be protected in advance with a known protecting group.
　例えば、市販されている（ＢＳＨ等として）、或いは公知の方法で容易に調製できる（下記合成例２８参照）、メルカプト基やアミノ基のような活性基を有するホウ素クラスターを用いるのであれば、その活性基に上記のＡの構築方法と同様にリンカー構造又はその断片構造を導入できる。また、活性基を有さないホウ素クラスターにリンカー構造又はその断片構造を直接導入する方法も知られており、例えば、下記スキームで表される方法が挙げられる。 For example, if a boron cluster having an active group such as a mercapto group or an amino group is used, which is commercially available (as BSH or the like) or can be easily prepared by a known method (see Synthesis Example 28 below), A linker structure or a fragment structure thereof can be introduced into the active group in the same manner as in the above construction method A. In addition, a method of directly introducing a linker structure or a fragment structure thereof into a boron cluster having no active group is also known, and examples thereof include a method represented by the following scheme.
Figure JPOXMLDOC01-appb-C000026
Figure JPOXMLDOC01-appb-C000026
［式中、Ｎｕは、求核付加反応により導入される基を示し、その他の記号は、上記定義の通りである。］ [Wherein Nu represents a group introduced by a nucleophilic addition reaction, and other symbols are as defined above. ]
　式（２）で表される化合物は、ホウ素クラスター（１）を、式（３）で表される化合物、酸（例えば、塩酸（塩化水素）等）、添加剤（例えば、テトラフルオロホウ酸ナトリウムのようなテトラフルオロホウ酸塩等）、必要に応じて溶媒（例えば、ジメチルスルホキシド（ＤＭＳＯ）のようなスルホキシド類、Ｎ－メチル－２－ピロリドン（ＮＭＰ）、Ｎ，Ｎ－ジメチルホルムアミド（ＤＭＦ）のようなアミド類、又はそれらの混合物）の存在下で反応させることにより得ることができる。反応温度は、通常－８０から１５０℃である。 The compound represented by the formula (2) is obtained by converting the boron cluster (1) into a compound represented by the formula (3), an acid (for example, hydrochloric acid (hydrogen chloride), etc.), an additive (for example, sodium tetrafluoroborate). Tetrafluoroborate, etc.), and optionally a solvent (for example, sulfoxides such as dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), N, N-dimethylformamide (DMF) Can be obtained by the reaction in the presence of amides such as The reaction temperature is usually from -80 to 150 ° C.
　式（４）で表される化合物は、式（２）で表される化合物を、対応する求核試薬、必要に応じて塩基（例えば、トリエチルアミンのような有機塩基、水酸化ナトリウム、水酸化カリウム、炭酸カリウム、フッ化セシウムのような無機塩基）の存在下で求核付加反応させることにより得ることができる。反応温度は、通常－８０から１５０℃である。その際、反応基質にカルボキシ基、ヒドロキシ基、アミノ基、メルカプト基のような活性基が存在する場合は、これらの官能基を、予め公知の保護基で保護しておいてもよい。反応温度は、通常－８０から１５０℃である。 The compound represented by the formula (4) is obtained by replacing the compound represented by the formula (2) with a corresponding nucleophile, optionally a base (for example, an organic base such as triethylamine, sodium hydroxide, potassium hydroxide). , Nucleophilic addition reaction in the presence of an inorganic base such as potassium carbonate or cesium fluoride). The reaction temperature is usually from -80 to 150 ° C. At that time, when an active group such as a carboxy group, a hydroxy group, an amino group, or a mercapto group is present on the reaction substrate, these functional groups may be protected in advance with a known protecting group. The reaction temperature is usually from -80 to 150 ° C.
　導入基に対応する求核試薬としては、アジド（－Ｎ３）基やアミノ（－ＮＨ２）基を導入する場合は、例えば、テトラブチルアンモニウムアジドが用いられ、中でもアミノ基を導入する場合は、反応後さらに、水素ガスのような還元剤及びパラジウム炭素のような触媒を用いる公知の還元反応に付す。反応温度は、通常－８０から１５０℃である。また、例えば、２－アミノエチルチオ（－Ｓ－（ＣＨ２）２－ＮＨ２）基を導入する場合は、ＨＳ－（ＣＨ２）２－ＮＨＢｏｃが用いられ、反応後に、Ｂｏｃ基を酸（例えば、塩酸、トリフルオロ酢酸等）で脱保護する。反応温度は、通常－８０から１５０℃である。 As the nucleophile corresponding to the introduction group, when introducing an azide (—N 3 ) group or an amino (—NH 2 ) group, for example, tetrabutylammonium azide is used, and in particular, when introducing an amino group. After the reaction, it is further subjected to a known reduction reaction using a reducing agent such as hydrogen gas and a catalyst such as palladium carbon. The reaction temperature is usually from -80 to 150 ° C. Further, for example, when introducing a 2-aminoethylthio (—S— (CH 2 ) 2 —NH 2 ) group, HS— (CH 2 ) 2 —NHBoc is used, and after the reaction, the Boc group is converted to an acid ( For example, deprotection is performed using hydrochloric acid, trifluoroacetic acid, or the like. The reaction temperature is usually from -80 to 150 ° C.
　式（５）で表される化合物は、式（２）で表される化合物を、チオ尿素、及び溶媒（例えば、メタノール、エタノール、ｔｅｒｔ－ブタノールのようなアルコール類）の存在下で反応させ、その後、塩基（例えば、ナトリウムメトキシド等）で処理することにより得ることができる。反応温度は、通常－８０から１５０℃である。 The compound represented by the formula (5) is obtained by reacting the compound represented by the formula (2) in the presence of thiourea and a solvent (for example, alcohols such as methanol, ethanol, tert-butanol), Then, it can obtain by processing with a base (for example, sodium methoxide etc.). The reaction temperature is usually from -80 to 150 ° C.
　化合物（ＩＩａ）、化合物（ＩＩｂ）のうち、リンカー構造又はその断片構造を導入する前の式（ＩＩａ’）及び（ＩＩｂ’）で表される化合物（以下、化合物（ＩＩａ’）、化合物（ＩＩｂ’）という）は、市販のものをそのまま用いてもよいし、公知のペプチド伸長法を用いて合成することもできる。 Among the compounds (IIa) and (IIb), the compounds represented by the formulas (IIa ′) and (IIb ′) (hereinafter referred to as the compound (IIa ′) and the compound (IIb) before introducing the linker structure or the fragment structure thereof. As for ()), a commercially available product may be used as it is, or it may be synthesized using a known peptide elongation method.
Figure JPOXMLDOC01-appb-C000027
Figure JPOXMLDOC01-appb-C000027
　例えば、Ｙ１又はＸ２がヒドロキシ基の場合は、Ｃ末端を固定するか或いは保護して、Ｙ１又はＸ２がヒドロキシ基以外の場合は、（Ｈ－）ｃＹ１又はＨ－Ｘ２で表される化合物等を出発原料として、公知のペプチド伸長法により、Ｃ末端からＮ末端にペプチド伸長反応を行う。ペプチド伸長の際、α－アミノ酸の側鎖や反応基質にカルボキシ基、ヒドロキシ基、アミノ基、メルカプト基のような活性基が存在する場合は、これらの官能基を、予め公知の保護基で保護しておいてもよい。伸長後、Ｘ１又はＹ２が水素原子以外の場合は、Ｘ１－Ｌ又はＹ２（－Ｌ）ｃ（式中、Ｌは脱離基又はＯＨ）で表される化合物と公知の縮合反応又は置換反応に付す。最後に、脱保護反応或いは必要に応じて各ペプチド伸長法で知られる後処理操作を行い、式（ＩＩａ）及び（ＩＩｂ）で表される化合物を得る。また、同様にＮ末端からＣ末端にペプチド伸長反応も行うこともできる。 For example, when Y 1 or X 2 is a hydroxy group, the C-terminus is fixed or protected, and when Y 1 or X 2 is other than a hydroxy group, (H—) c Y 1 or H—X 2 As a starting material, a peptide elongation reaction is performed from the C-terminus to the N-terminus by a known peptide elongation method. During peptide elongation, if active groups such as carboxy group, hydroxy group, amino group, mercapto group are present in the side chain of α-amino acid or reaction substrate, these functional groups are protected with known protecting groups in advance. You may keep it. After extension, when X 1 or Y 2 is other than a hydrogen atom, a compound represented by X 1 -L or Y 2 (-L) c (wherein L is a leaving group or OH) and a known condensation reaction Or it attaches to substitution reaction. Finally, a deprotection reaction or, if necessary, a post-treatment operation known for each peptide extension method is performed to obtain compounds represented by the formulas (IIa) and (IIb). Similarly, a peptide extension reaction can also be performed from the N-terminus to the C-terminus.
　ペプチド伸長法としては、例えば、活性化エステル法、混合酸無水物法、アジド法等のＣ末端活性化法、カルボジイミド等のカップリング法、Ｎ－カルボン酸無水物（ＮＣＡ）法、酸化還元法、酵素法、固相合成法等が挙げられる。 Examples of peptide elongation methods include activated ester method, mixed acid anhydride method, C-terminal activation method such as azide method, coupling method such as carbodiimide, N-carboxylic acid anhydride (NCA) method, redox method Enzyme method, solid phase synthesis method and the like.
　以下に、合成例、比較例、実施例及び試験例に基づいて本発明をより詳細に説明するが、本発明はこれらにより限定されるものではなく、また本発明の範囲を逸脱しない範囲で変化させてもよい。 Hereinafter, the present invention will be described in more detail based on synthesis examples, comparative examples, examples, and test examples, but the present invention is not limited thereto, and changes may be made without departing from the scope of the present invention. You may let them.
　以下、化学構造式や標題、文章中における重合度は、おおよその平均重合度を示す。また、化学構造式内のポリエーテルの重合度ｎは各合成例及び実施例ごとに独立した値を有する。本明細書中「１０Ｂ－」は、１０Ｂの同位体比が９８％以上である化合物を示し、「天然型Ｂ－」は、天然の同位体比（８０．１％の１１Ｂと１９．９％の１０Ｂ）である化合物を示す。 Hereinafter, the degree of polymerization in the chemical structural formula, title, and text indicates an approximate average degree of polymerization. Further, the polymerization degree n of the polyether in the chemical structural formula has an independent value for each synthesis example and each example. In the present specification, “ 10 B-” indicates a compound having an isotope ratio of 10 B of 98% or more, and “natural B-” indicates a natural isotope ratio ( 11 B and 19 of 80.1%). 9% of 10 B).
合成例１
ポリエチレングリコール（Ｍｎ＝１２，０００）－ポリグルタミン酸（４０ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ））の合成 Synthesis example 1
Synthesis of polyethylene glycol (Mn = 12,000) -polyglutamic acid (40mer) -block copolymer (PEG-b-P (Glu))
Figure JPOXMLDOC01-appb-C000028
Figure JPOXMLDOC01-appb-C000028
　アルゴン雰囲気下、γ－ベンジル Ｌ－グルタメート Ｎ－カルボン酸無水物（ＢＬＧ－ＮＣＡ，５２６　ｍｇ，２．０　ｍｍｏｌ）をＤＭＦ（２６　ｍｌ）に溶解した後、ＤＭＦ（３０　ｍｌ）に溶解させたα－メトキシ－ω－アミノ－ポリエチレングルコール（ＣＨ３Ｏ－ＰＥＧ－ＮＨ２，Ｍｎ＝１２，０００，６００　ｍｇ，０．０５　ｍｍｏｌ）を加え、３５℃にて２日間撹拌することで開環重合を行った。ジエチルエーテル中で沈殿を生成させ、その沈殿をろ取し、真空乾燥することで、ＰＥＧ－ｂ－Ｐ（Ｇｌｕ）のベンジル保護体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ（Ｂｎ））を得た。ポリアミノ酸の重合度（４０ｍｅｒ）は、１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６，８０℃）において、ＰＥＧ部分のプロトンの積分値（－ＯＣＨ ２ ＣＨ ２ －，δ＝３．６　ｐｐｍ）とポリグルタミン酸ベンジルエステルの芳香環部分の積分値（Ｃ６ Ｈ ５ ＣＨ２－，δ＝７．３　ｐｐｍ）を比較することで、算出した。分子量分布は、ゲルろ過クロマトグラフィー（ＧＰＣ）分析（カラム：ＴＳＫ－ｇｅｌ　Ｇ３０００ＨＨＲ，Ｇ４０００ＨＨＲ，東ソー株式会社，溶離液：１０　ｍＭ　ＬｉＣｌのＤＭＦ溶液，流速：０．８　ｍｌ/ｍｉｎ，検出：示差屈折率（ＲＩ）検出器；温度：２５°Ｃ）から、１．０３と算出した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６，８０℃）δ（ｐｐｍ）： ８．２（ｓ，４０Ｈ），７．３（ｍ，２００Ｈ），５．１（ｍ，８０Ｈ），４．０（ｓ，４０Ｈ），３．６（ｍ，１０９０Ｈ），２．１－２．５（ｍ，１６０Ｈ）．
　こうして得られたＰＥＧ－ｂ－Ｐ（Ｇｌｕ（Ｂｎ））を０．５Ｎ水酸化ナトリウム水溶液で処理してベンジル基を脱保護後、透析、凍結乾燥を行って標題化合物を得た。 Under an argon atmosphere, γ-benzyl L-glutamate N-carboxylic anhydride (BLG-NCA, 526 mg, 2.0 mmol) was dissolved in DMF (26 ml) and then dissolved in DMF (30 ml). -Methoxy-ω-amino-polyethylene glycol (CH 3 O-PEG-NH 2 , Mn = 12,000,600 mg, 0.05 mmol) was added, and the mixture was stirred at 35 ° C. for 2 days for ring-opening polymerization Went. A precipitate was generated in diethyl ether, and the precipitate was collected by filtration and dried under vacuum to obtain a benzyl-protected form of PEG-b-P (Glu) (PEG-b-P (Glu (Bn))). polymerization degree of polyamino acid (40 mer) is, 1 H-NMR (400 MHz , DMSO- d 6, 80 ℃) at an integrated value of the PEG portion of the proton (-OC H 2 C H 2 - , δ = 3.6 ppm) and the integral value (C 6 H 5 CH 2 —, δ = 7.3 ppm) of the aromatic ring portion of polyglutamic acid benzyl ester, and the molecular weight distribution was calculated by gel filtration chromatography (GPC). analysis (column: TSK-gel G3000 HHR, G4000 HHR, Tosoh Corporation, eluent: DMF solution of 10 mM LiCl, flow rate: 0.8 ml / min, detection: differential refractive Rate (RI) detector; Temperature: from 25 ° C), was calculated as 1.03.
1 H-NMR (400 MHz, DMSO-d 6 , 80 ° C.) δ (ppm): 8.2 (s, 40 H), 7.3 (m, 200 H), 5.1 (m, 80 H), 4. 0 (s, 40H), 3.6 (m, 1090H), 2.1-2.5 (m, 160H).
The PEG-bP (Glu (Bn)) thus obtained was treated with a 0.5N aqueous sodium hydroxide solution to remove the benzyl group, followed by dialysis and lyophilization to obtain the title compound.
合成例２
ポリエチレングリコール（Ｍｎ＝１２，０００）－チオレート化ポリグルタミン酸（４０ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｔｈｉｏｌａｔｅ））の合成 Synthesis example 2
Synthesis of polyethylene glycol (Mn = 12,000) -thiolated polyglutamic acid (40mer) -block copolymer (PEG-bP (Glu-Thiolate))
Figure JPOXMLDOC01-appb-C000029
Figure JPOXMLDOC01-appb-C000029
　合成例１で得られた化合物（３００　ｍｇ）に対して、０．０１Ｎ塩酸中で透析を行うことで、ポリグルタミン酸の対カチオンをナトリウムカチオンからプロトンへとイオン交換した後、凍結乾燥を行った。得られた化合物、ＥＤＣ（１４７．４　ｍｇ，１．１　ｅｑ）、Ｎ－ヒドロキシスクシンイミド（ＮＨＳ）（８８．５　ｍｇ，１．１　ｅｑ）をＤＭＳＯ（１０　ｍｌ）中で３０分撹拌した後、２－（ピリジニルジスルファニル）エタンアミン塩酸塩（１５６．５　ｍｇ，１．０　ｅｑ）を加え、さらに２４時間撹拌した。反応終了後、この混合物を水中で透析し、凍結乾燥を行って、標題化合物を得た。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６，８０℃）δ（ｐｐｍ）： ８．２（ｓ，４０Ｈ），７．７（ｍ，８０Ｈ），４．０（ｓ，４０Ｈ），３．６（ｍ，１０９０Ｈ），２．１－２．５（ｍ，１６０Ｈ）． The compound (300 mg) obtained in Synthesis Example 1 was dialyzed in 0.01N hydrochloric acid to ion-exchange a polyglutamic acid counter cation from a sodium cation to a proton, and then lyophilized. . The obtained compound, EDC (147.4 mg, 1.1 eq), N-hydroxysuccinimide (NHS) (88.5 mg, 1.1 eq) was stirred in DMSO (10 ml) for 30 minutes. 2- (Pyridinyldisulfanyl) ethanamine hydrochloride (156.5 mg, 1.0 eq) was added, and the mixture was further stirred for 24 hours. After completion of the reaction, the mixture was dialyzed in water and lyophilized to give the title compound.
1 H-NMR (400 MHz, DMSO-d 6 , 80 ° C.) δ (ppm): 8.2 (s, 40H), 7.7 (m, 80H), 4.0 (s, 40H), 3. 6 (m, 1090H), 2.1-2.5 (m, 160H).
実施例１
ポリエチレングリコール（Ｍｎ＝１２，０００）－ＢＳＨ結合ポリグルタミン酸（４０ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－ＢＳＨ））の合成 Example 1
Synthesis of polyethylene glycol (Mn = 12,000) -BSH-linked polyglutamic acid (40mer) -block copolymer (PEG-bP (Glu-BSH))
Figure JPOXMLDOC01-appb-C000030
Figure JPOXMLDOC01-appb-C000030
　合成例２で得られた化合物（１０　ｍｇ）をＤＭＳＯ（１０　ｍｌ）に溶解し、それをメタノール（２　ｍｌ）に溶解した１０Ｂ－ＢＳＨ（４．２　ｍｇ，ブロック共重合体中のチオレート化された官能基に対して２　ｅｑ）と混合して、室温で終夜反応させた後、反応液を薄い水酸化ナトリウム水溶液中で透析し、凍結乾燥することで、標題化合物を得た。標題化合物（１　ｍｇ）を９０％硝酸（１　ｍｌ）に溶解後、１％硝酸で希釈した溶液に対してＩＣＰ－ＭＳ分析を行い、１０Ｂ量を定量することによって、ポリマー１分子に対して約２０個の１０Ｂ－ＢＳＨが導入されたこと、ポリマー１分子当りの１０Ｂ－ＢＳＨ導入量は１８．７％（ｗ／ｗ）であることを決定した。 The compound (10 mg) obtained in Synthesis Example 2 was dissolved in DMSO (10 ml), and 10 B-BSH (4.2 mg, thiolated in a block copolymer) dissolved in methanol (2 ml). After mixing with 2 eq) for the functional group and reacting overnight at room temperature, the reaction solution was dialyzed in a thin aqueous sodium hydroxide solution and freeze-dried to obtain the title compound. After dissolving the title compound (1 mg) in 90% nitric acid (1 ml), subjected to ICP-MS analysis on the solution diluted with 1% nitric acid, by quantifying the 10 B amount, relative to the polymer molecule It was determined that about 20 10 B-BSH had been introduced, and the amount of 10 B-BSH introduced per polymer molecule was 18.7% (w / w).
合成例３
ポリエチレングリコール（Ｍｎ＝１２，０００）－ポリグルタミン酸（９０ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ））の合成 Synthesis example 3
Synthesis of polyethylene glycol (Mn = 12,000) -polyglutamic acid (90mer) -block copolymer (PEG-b-P (Glu))
Figure JPOXMLDOC01-appb-C000031
Figure JPOXMLDOC01-appb-C000031
　合成例１と同様の方法で、α－メトキシ－ω－アミノ－ポリエチレングルコール（ＣＨ３Ｏ－ＰＥＧ－ＮＨ２，Ｍｎ＝１２，０００，１２０　ｍｇ，０．０１　ｍｍｏｌ）とＢＬＧ－ＮＣＡ（２３７　ｍｇ，０．９　ｍｍｏｌ）による開環重合を行い、ＰＥＧ－ｂ－Ｐ（Ｇｌｕ（Ｂｎ））を得た。ポリアミノ酸の重合度（９０ｍｅｒ）は、合成例１と同様の方法で算出した。分子量分布は、合成例１と同様の方法で、１．１５と算出した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６，８０℃）δ（ｐｐｍ）：８．２（ｓ，９０Ｈ），７．３（ｍ，４５０Ｈ），５．１（ｍ，１８０Ｈ），４．０（ｓ，９０Ｈ），３．６（ｍ，１０９０Ｈ），２．１－２．５（ｍ，３６０Ｈ）．
　得られた化合物に対して、合成例１と同様の方法で脱保護を行い、ポリエチレングリコール（Ｍｎ＝１２，０００）－ポリグルタミン酸（９０ｍｅｒ）－ブロック共重合体を得た。 In the same manner as in Synthesis Example 1, α-methoxy-ω-amino-polyethylene glycol (CH 3 O-PEG-NH 2 , Mn = 12,000, 120 mg, 0.01 mmol) and BLG-NCA (237 mg, 0.9 mmol) was subjected to ring-opening polymerization to obtain PEG-b-P (Glu (Bn)). The degree of polymerization of polyamino acid (90 mer) was calculated in the same manner as in Synthesis Example 1. The molecular weight distribution was calculated to be 1.15 by the same method as in Synthesis Example 1.
1 H-NMR (400 MHz, DMSO-d 6 , 80 ° C.) δ (ppm): 8.2 (s, 90 H), 7.3 (m, 450 H), 5.1 (m, 180 H), 4. 0 (s, 90H), 3.6 (m, 1090H), 2.1-2.5 (m, 360H).
The resulting compound was deprotected in the same manner as in Synthesis Example 1 to obtain polyethylene glycol (Mn = 12,000) -polyglutamic acid (90mer) -block copolymer.
合成例４
ポリエチレングリコール（Ｍｎ＝１２，０００）－チオレート化ポリグルタミン酸（９０ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｔｈｉｏｌａｔｅ））の合成 Synthesis example 4
Synthesis of polyethylene glycol (Mn = 12,000) -thiolated polyglutamic acid (90mer) -block copolymer (PEG-bP (Glu-Thiolate))
Figure JPOXMLDOC01-appb-C000032
Figure JPOXMLDOC01-appb-C000032
　合成例３で得られた化合物から、合成例２と同様の方法で、ポリエチレングリコール（Ｍｎ＝１２，０００）－チオレート化ポリグルタミン酸（９０ｍｅｒ）－ブロック共重合体を合成した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６，８０℃）δ（ｐｐｍ）：８．２（ｓ，９０Ｈ），７．７（ｍ，１９２Ｈ），４．０（ｓ，９０Ｈ），３．６（ｍ，１０９０Ｈ），２．１－２．５（ｍ，３６０Ｈ）． From the compound obtained in Synthesis Example 3, a polyethylene glycol (Mn = 12,000) -thiolated polyglutamic acid (90mer) -block copolymer was synthesized in the same manner as in Synthesis Example 2.
1 H-NMR (400 MHz, DMSO-d 6 , 80 ° C.) δ (ppm): 8.2 (s, 90 H), 7.7 (m, 192 H), 4.0 (s, 90 H), 3. 6 (m, 1090H), 2.1-2.5 (m, 360H).
実施例２
ポリエチレングリコール（Ｍｎ＝１２，０００）－ＢＳＨ結合ポリグルタミン酸（９０ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－ＢＳＨ））の合成 Example 2
Synthesis of polyethylene glycol (Mn = 12,000) -BSH-linked polyglutamic acid (90mer) -block copolymer (PEG-bP (Glu-BSH))
Figure JPOXMLDOC01-appb-C000033
Figure JPOXMLDOC01-appb-C000033
　合成例４で得られた化合物に対して、実施例１と同様の方法で、１０Ｂ－ＢＳＨと縮合して、標題化合物を合成した。実施例１と同様の方法で、ポリマー１分子に対して約４８個の１０Ｂ－ＢＳＨが導入されたこと、ポリマー１分子当りの１０Ｂ－ＢＳＨ導入量は２７．６％（ｗ／ｗ）であることを決定した。 The title compound was synthesized by condensing the compound obtained in Synthesis Example 4 with 10 B-BSH in the same manner as in Example 1. In the same manner as in Example 1, about 48 10 B-BSH was introduced per molecule of polymer, and the introduced amount of 10 B-BSH per polymer molecule was 27.6% (w / w). Decided to.
合成例５
ポリエチレングリコール（Ｍｎ＝２０，０００）－ポリグルタミン酸（４０ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ））の合成 Synthesis example 5
Synthesis of polyethylene glycol (Mn = 20,000) -polyglutamic acid (40mer) -block copolymer (PEG-b-P (Glu))
Figure JPOXMLDOC01-appb-C000034
Figure JPOXMLDOC01-appb-C000034
　合成例１と同様の方法で、ＣＨ３Ｏ－ＰＥＧ－ＮＨ２（Ｍｎ＝１２，０００）の代わりにＣＨ３Ｏ－ＰＥＧ－ＮＨ２（Ｍｎ＝２０，０００，２００　ｍｇ，０．０１　ｍｍｏｌ）を用い、ＢＬＧ－ＮＣＡ（１０６　ｍｇ，０．４　ｍｍｏｌ）による開環重合を行い、ＰＥＧ－ｂ－Ｐ（Ｇｌｕ（Ｂｎ））を得た。ポリアミノ酸の重合度（４０ｍｅｒ）は、合成例１と同様の方法で算出した。分子量分布は、合成例１と同様の方法で、１．０５と算出した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６，８０℃）δ（ｐｐｍ）：８．２（ｓ，４０Ｈ），７．３（ｍ，２００Ｈ），５．１（ｍ，８０Ｈ），４．０（ｓ，４０Ｈ），３．６（ｍ，１８１８Ｈ），２．１－２．５（ｍ，１６０Ｈ）．
　得られた化合物に対して、合成例１と同様の方法で脱保護を行い、ポリエチレングリコール（Ｍｎ＝２０，０００）－ポリグルタミン酸（４０ｍｅｒ）－ブロック共重合体を合成した。 In the same manner as in Synthesis Example 1, CH 3 O-PEG- NH 2 instead of CH 3 O-PEG-NH 2 (Mn = 12,000) (Mn = 20,000,200 mg, 0.01 mmol) Was used for ring-opening polymerization with BLG-NCA (106 mg, 0.4 mmol) to obtain PEG-b-P (Glu (Bn)). The polymerization degree (40 mer) of the polyamino acid was calculated in the same manner as in Synthesis Example 1. The molecular weight distribution was calculated to be 1.05 by the same method as in Synthesis Example 1.
1 H-NMR (400 MHz, DMSO-d 6 , 80 ° C.) δ (ppm): 8.2 (s, 40 H), 7.3 (m, 200 H), 5.1 (m, 80 H), 4. 0 (s, 40H), 3.6 (m, 1818H), 2.1-2.5 (m, 160H).
The obtained compound was deprotected in the same manner as in Synthesis Example 1 to synthesize a polyethylene glycol (Mn = 20,000) -polyglutamic acid (40mer) -block copolymer.
合成例６
ポリエチレングリコール（Ｍｎ＝２０，０００）－チオレート化ポリグルタミン酸（４０ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｔｈｉｏｌａｔｅ））の合成 Synthesis Example 6
Synthesis of polyethylene glycol (Mn = 20,000) -thiolated polyglutamic acid (40mer) -block copolymer (PEG-bP (Glu-Thiolate))
Figure JPOXMLDOC01-appb-C000035
Figure JPOXMLDOC01-appb-C000035
　合成例５で得られた化合物から、合成例２と同様の方法で、ポリエチレングリコール（Ｍｎ＝２０，０００）－チオレート化ポリグルタミン酸（４０ｍｅｒ）－ブロック共重合体を合成した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６，８０℃）δ（ｐｐｍ）：８．２（ｓ，４０Ｈ），７．７（ｍ，８０Ｈ），４．０（ｓ，４０Ｈ），３．６（ｍ，１８１８Ｈ），２．１－２．５（ｍ，１６０Ｈ）． Polyethylene glycol (Mn = 20,000) -thiolated polyglutamic acid (40mer) -block copolymer was synthesized from the compound obtained in Synthesis Example 5 in the same manner as in Synthesis Example 2.
1 H-NMR (400 MHz, DMSO-d 6 , 80 ° C.) δ (ppm): 8.2 (s, 40H), 7.7 (m, 80H), 4.0 (s, 40H), 3. 6 (m, 1818H), 2.1-2.5 (m, 160H).
実施例３
ポリエチレングリコール（Ｍｎ＝２０，０００）－ＢＳＨ結合ポリグルタミン酸（４０ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－ＢＳＨ））の合成 Example 3
Synthesis of polyethylene glycol (Mn = 20,000) -BSH-linked polyglutamic acid (40mer) -block copolymer (PEG-bP (Glu-BSH))
Figure JPOXMLDOC01-appb-C000036
Figure JPOXMLDOC01-appb-C000036
　合成例６で得られた化合物に対して、実施例１と同様の方法で、１０Ｂ－ＢＳＨと縮合して、標題化合物を合成した。実施例１と同様の方法で、ポリマー１分子に対して約２０個の１０Ｂ－ＢＳＨが導入されたこと、ポリマー１分子当りの１０Ｂ－ＢＳＨ導入量は１３．８％（ｗ／ｗ）であることを決定した。 The title compound was synthesized by condensing the compound obtained in Synthesis Example 6 with 10 B-BSH in the same manner as in Example 1. In the same manner as in Example 1, about 20 10 B-BSH was introduced per molecule of polymer, and the introduced amount of 10 B-BSH per polymer molecule was 13.8% (w / w). Decided to.
合成例７
ポリグルタミン酸（４０ｍｅｒ）（Ｐ（Ｇｌｕ））の合成 Synthesis example 7
Synthesis of polyglutamic acid (40mer) (P (Glu))
Figure JPOXMLDOC01-appb-C000037
Figure JPOXMLDOC01-appb-C000037
　合成例１と同様の方法で、ＣＨ３Ｏ－ＰＥＧ－ＮＨ２の代わりにｎ－ブチルアミン（２　ｍｇ，０．０２７　ｍｍｏｌ）を用い、ＢＬＧ－ＮＣＡ（２８９　ｍｇ，１．０８　ｍｍｏｌ）による開環重合を行い、ポリグルタミン酸のベンジル保護体Ｐ（Ｇｌｕ（Ｂｎ））（４０ｍｅｒ）を得た。ポリアミノ酸の重合度（４０ｍｅｒ）は、１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６，８０℃）において、ｎ－ブチル基末端のメチル基部分のプロトンの積分値（－ＣＨ 3 ，δ＝１．４　ｐｐｍ，１．６　ｐｐｍ）とポリグルタミン酸ベンジルエステルの芳香環部分の積分値（Ｃ６ Ｈ ５ ＣＨ２－，δ＝７．３　ｐｐｍ）を比較することで、算出した。分子量分布は、合成例１と同様の方法で、１．１５と算出した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６，８０℃）δ（ｐｐｍ）：８．２（ｓ，４０Ｈ），７．３（ｍ，２００Ｈ），５．１（ｍ，８０Ｈ），４．０（ｓ，４０Ｈ），２．１－２．５（ｍ，１６０Ｈ），０．８（ｔ，３Ｈ）．
　得られた化合物に対して、合成例１と同様の方法で脱保護を行い、ホモポリマーのポリグルタミン酸（４０ｍｅｒ）を合成した。 Ring opening with BLG-NCA (289 mg, 1.08 mmol) using n-butylamine (2 mg, 0.027 mmol) instead of CH 3 O-PEG-NH 2 in the same manner as in Synthesis Example 1. Polymerization was performed to obtain a polyglutamic acid benzyl protector P (Glu (Bn)) (40mer). Polymerization degree of polyamino acid (40 mer) is, 1 H-NMR in (400 MHz, DMSO-d 6 , 80 ℃), the integral value of the proton of methyl group portion of the n- butyl terminated (-C H 3, δ = 1.4 ppm, 1.6 ppm) and the integrated value (C 6 H 5 CH 2 —, δ = 7.3 ppm) of the aromatic ring portion of polyglutamic acid benzyl ester were calculated. The molecular weight distribution was calculated to be 1.15 by the same method as in Synthesis Example 1.
1 H-NMR (400 MHz, DMSO-d 6 , 80 ° C.) δ (ppm): 8.2 (s, 40 H), 7.3 (m, 200 H), 5.1 (m, 80 H), 4. 0 (s, 40H), 2.1-2.5 (m, 160H), 0.8 (t, 3H).
The obtained compound was deprotected in the same manner as in Synthesis Example 1 to synthesize a homopolymer polyglutamic acid (40mer).
合成例８
チオレート化ポリグルタミン酸（４０ｍｅｒ）（Ｐ（Ｇｌｕ－Ｔｈｉｏｌａｔｅ））の合成 Synthesis example 8
Synthesis of thiolated polyglutamic acid (40mer) (P (Glu-Thiolate))
Figure JPOXMLDOC01-appb-C000038
Figure JPOXMLDOC01-appb-C000038
　合成例７で得られた化合物を用いて、合成例２と同様の方法で、チオレート化反応を行った。反応終了後、この混合物をメタノール中で透析し、最後にベンゼン／メタノール（９／１（ｖ／ｖ））溶液として凍結乾燥を行って、チオレート化ポリグルタミン酸（４０ｍｅｒ）を合成した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６，８０℃）δ（ｐｐｍ）：８．２（ｓ，４０Ｈ），７．７（ｍ，７６Ｈ），４．０（ｓ，４０Ｈ），２．１－２．５（ｍ，１６０Ｈ），０．８（ｔ，３Ｈ）． Using the compound obtained in Synthesis Example 7, a thiolation reaction was performed in the same manner as in Synthesis Example 2. After completion of the reaction, the mixture was dialyzed in methanol, and finally lyophilized as a benzene / methanol (9/1 (v / v)) solution to synthesize thiolated polyglutamic acid (40mer).
1 H-NMR (400 MHz, DMSO-d 6 , 80 ° C.) δ (ppm): 8.2 (s, 40H), 7.7 (m, 76H), 4.0 (s, 40H), 2. 1-2.5 (m, 160H), 0.8 (t, 3H).
比較例１
ＢＳＨ結合ポリグルタミン酸（４０ｍｅｒ）（Ｐ（Ｇｌｕ－ＢＳＨ））の合成 Comparative Example 1
Synthesis of BSH-linked polyglutamic acid (40mer) (P (Glu-BSH))
Figure JPOXMLDOC01-appb-C000039
Figure JPOXMLDOC01-appb-C000039
　合成例８で得られた化合物に対して、実施例１と同様の方法で、１０Ｂ－ＢＳＨと縮合して、標題化合物を合成した。実施例１と同様の方法で、ポリマー１分子に対して約１９個の１０Ｂ－ＢＳＨが導入されたこと、ポリマー１分子当りの１０Ｂ－ＢＳＨ導入量は３８．６％（ｗ／ｗ）であることを決定した。 The title compound was synthesized by condensing the compound obtained in Synthesis Example 8 with 10 B-BSH in the same manner as in Example 1. In the same manner as in Example 1, about 19 10 B-BSH was introduced per molecule of polymer, and the introduced amount of 10 B-BSH per molecule was 38.6% (w / w). Decided to.
合成例９
ポリグルタミン酸（９０ｍｅｒ）（Ｐ（Ｇｌｕ））の合成 Synthesis Example 9
Synthesis of polyglutamic acid (90mer) (P (Glu))
Figure JPOXMLDOC01-appb-C000040
Figure JPOXMLDOC01-appb-C000040
　合成例１と同様の方法で、ＣＨ３Ｏ－ＰＥＧ－ＮＨ２の代わりにｎ－ブチルアミン（２　ｍｇ，０．０２７　ｍｍｏｌ）を用い、ＢＬＧ－ＮＣＡ（６５０　ｍｇ，２．４３　ｍｍｏｌ）による開環重合を行い、ポリグルタミン酸のベンジル保護体Ｐ（Ｇｌｕ（Ｂｎ））（９０ｍｅｒ）を得た。ポリアミノ酸の重合度（９０ｍｅｒ）は、１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６，８０℃）において、ｎ－ブチル基末端のメチル基部分のプロトンの積分値（－ＣＨ ３ ，δ＝０．８　ｐｐｍ）とポリグルタミン酸ベンジルエステルの芳香環部分の積分値（Ｃ６ Ｈ ５ ＣＨ２－，δ＝７．３　ｐｐｍ）を比較することで、算出した。分子量分布は、合成例１と同様の方法で、１．１５と算出した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６，８０℃）δ（ｐｐｍ）：８．２（ｓ，９０Ｈ），７．３（ｍ，４５０Ｈ），５．１（ｍ，１８０Ｈ），４．０（ｓ，９０Ｈ），２．１－２．５（ｍ，３６０Ｈ），０．８（ｔ，３Ｈ）．
　得られた化合物に対して、合成例１と同様の方法で脱保護を行い、ホモポリマーのポリグルタミン酸（９０ｍｅｒ）を合成した。 Ring opening with BLG-NCA (650 mg, 2.43 mmol) using n-butylamine (2 mg, 0.027 mmol) instead of CH 3 O-PEG-NH 2 in the same manner as in Synthesis Example 1. Polymerization was carried out to obtain a polyglutamic acid benzyl protector P (Glu (Bn)) (90mer). Polymerization degree of polyamino acid (90 mer) is, 1 H-NMR in (400 MHz, DMSO-d 6 , 80 ℃), the integral value of the proton of methyl group portion of the n- butyl terminated (-C H 3, δ = 0.8 ppm) and the integral value (C 6 H 5 CH 2 —, δ = 7.3 ppm) of the aromatic ring portion of polyglutamic acid benzyl ester was calculated. The molecular weight distribution was calculated to be 1.15 by the same method as in Synthesis Example 1.
1 H-NMR (400 MHz, DMSO-d 6 , 80 ° C.) δ (ppm): 8.2 (s, 90 H), 7.3 (m, 450 H), 5.1 (m, 180 H), 4. 0 (s, 90H), 2.1-2.5 (m, 360H), 0.8 (t, 3H).
The resulting compound was deprotected in the same manner as in Synthesis Example 1 to synthesize a homopolymer polyglutamic acid (90mer).
合成例１０
チオレート化ポリグルタミン酸（９０ｍｅｒ）（Ｐ（Ｇｌｕ－Ｔｈｉｏｌａｔｅ））の合成 Synthesis Example 10
Synthesis of thiolated polyglutamic acid (90mer) (P (Glu-Thiolate))
Figure JPOXMLDOC01-appb-C000041
Figure JPOXMLDOC01-appb-C000041
　合成例９で得られた化合物を用いて、合成例２と同様の方法で、チオレート化反応を行った。反応終了後、この混合物をメタノール中で透析し、最後にベンゼン／メタノール（９／１（ｖ／ｖ））溶液として凍結乾燥を行って、チオレート化ポリグルタミン酸（９０ｍｅｒ）を合成した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６，８０℃）δ（ｐｐｍ）：８．２（ｓ，９０Ｈ），７．７（ｍ，１６４Ｈ），４．０（ｓ，９０Ｈ），２．１－２．５（ｍ，７２０Ｈ），０．８（ｔ，３Ｈ）． Using the compound obtained in Synthesis Example 9, a thiolation reaction was performed in the same manner as in Synthesis Example 2. After completion of the reaction, the mixture was dialyzed in methanol, and finally lyophilized as a benzene / methanol (9/1 (v / v)) solution to synthesize thiolated polyglutamic acid (90mer).
1 H-NMR (400 MHz, DMSO-d 6 , 80 ° C.) δ (ppm): 8.2 (s, 90 H), 7.7 (m, 164 H), 4.0 (s, 90 H), 2. 1-2.5 (m, 720H), 0.8 (t, 3H).
比較例２
ＢＳＨ結合ポリグルタミン酸（９０ｍｅｒ）（Ｐ（Ｇｌｕ－ＢＳＨ））の合成 Comparative Example 2
Synthesis of BSH-linked polyglutamic acid (90mer) (P (Glu-BSH))
Figure JPOXMLDOC01-appb-C000042
Figure JPOXMLDOC01-appb-C000042
　合成例１０で得られた化合物に対して、実施例１と同様の方法で、１０Ｂ－ＢＳＨと縮合して、標題化合物を合成した。実施例１と同様の方法で、ポリマー１分子に対して約４１個の１０Ｂ－ＢＳＨが導入されたこと、ポリマー１分子当りの１０Ｂ－ＢＳＨ導入量は３８．０％（ｗ／ｗ）であることを決定した。 The title compound was synthesized by condensing the compound obtained in Synthesis Example 10 with 10 B-BSH in the same manner as in Example 1. In the same manner as in Example 1, about 41 10 B-BSH was introduced per molecule of polymer, and the introduced amount of 10 B-BSH per polymer molecule was 38.0% (w / w). Decided to.
合成例１１
ポリエチレングリコール（Ｍｗ＝１０，０００）－ベンジル保護ポリアスパラギン酸（２３ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ａｓｐ（Ｂｎ）））の合成 Synthesis Example 11
Synthesis of polyethylene glycol (Mw = 10,000) -benzyl-protected polyaspartic acid (23mer) -block copolymer (PEG-bP (Asp (Bn)))
Figure JPOXMLDOC01-appb-C000043
Figure JPOXMLDOC01-appb-C000043
　アルゴン雰囲気下にて、Ｈ２Ｎ－ＰＥＧ－ＯＭｅ（Ｍｗ＝１０，０００，３２３　ｍｇ，０．０３２３　ｍｍｏｌ）をジクロロメタン（１２．５　ｍｌ）に溶解させ、ＤＭＦ（３．０　ｍｌ）に溶解させたβ－ベンジル Ｌ－アスパルテート Ｎ－カルボン酸無水物（ＢＬＡ－ＮＣＡ）（２００　ｍｇ，０．８０３　ｍｍｏｌ）を滴下し、４０℃で１６時間攪拌した。室温にて、反応液をヘキサン／酢酸エチル（３／２（ｖ／ｖ），５０　ｍｌ）中に滴下して再沈殿させ、標題化合物（３９２ｍｇ，０．０２９５　ｍｍｏｌ，収率９１％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．２９、１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６）により、得られた化合物の重合度が２３であることを確認した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６）δ（ｐｐｍ）：２．５４－２．７１（ｍ，２３Ｈ），２．７６－２．９０（ｍ，２３Ｈ），３．３８－３．６１（ｍ，９０９Ｈ），４．５３－４．６９（ｍ，２１Ｈ），４．９６－５．１０（ｍ，４６Ｈ），７．１８－７．４１（ｍ，１１７Ｈ）． Under an argon atmosphere, H 2 N-PEG-OMe (Mw = 10,000, 323 mg, 0.0323 mmol) was dissolved in dichloromethane (12.5 ml) and dissolved in DMF (3.0 ml). Β-Benzyl L-aspartate N-carboxylic anhydride (BLA-NCA) (200 mg, 0.803 mmol) was added dropwise and stirred at 40 ° C. for 16 hours. At room temperature, the reaction solution was re-precipitated dropwise in hexane / ethyl acetate (3/2 (v / v), 50 ml) to give the title compound (392 mg, 0.0295 mmol, yield 91%) as white. Obtained as a powder. It was confirmed by size exclusion chromatography that the molecular weight distribution (Mw / Mn) was 1.29 and the degree of polymerization of the obtained compound was 23 by 1 H-NMR (400 MHz, DMSO-d 6 ).
1 H-NMR (400 MHz, DMSO-d 6 ) δ (ppm): 2.54-2.71 (m, 23H), 2.76-2.90 (m, 23H), 3.38-3. 61 (m, 909H), 4.53-4.69 (m, 21H), 4.96-5.10 (m, 46H), 7.18-7.41 (m, 117H).
合成例１２
ポリエチレングリコール（Ｍｗ＝１０，０００）－ポリアスパラギン酸（２３ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ａｓｐ））の合成 Synthesis Example 12
Synthesis of polyethylene glycol (Mw = 10,000) -polyaspartic acid (23mer) -block copolymer (PEG-bP (Asp))
Figure JPOXMLDOC01-appb-C000044
Figure JPOXMLDOC01-appb-C000044
　合成例１１で得られた化合物（３０９　ｍｇ，０．０２３３　ｍｍｏｌ）をＮ－メチルピロリドン（ＮＭＰ）／Ｈ２Ｏ（１／１（ｖ／ｖ），１２．０　ｍｌ）に溶解させ、１Ｎ水酸化ナトリウム水溶液（２．０　ｍｌ）を氷冷下で滴下し、室温にて２時間攪拌した。この溶液を、脱イオン水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（２０７　ｍｇ，０．０１６６　ｍｍｏｌ，収率７３％）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）により、脱保護が完了したことを確認した。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．０６であった。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：２．４１－２．８７（ｍ，４４Ｈ），３．５２－３，７８（ｍ，９０９Ｈ），４．３０－４．５３（ｍ，１６Ｈ）． The compound obtained in Synthesis Example 11 (309 mg, 0.0233 mmol) was dissolved in N-methylpyrrolidone (NMP) / H 2 O (1/1 (v / v), 12.0 ml), and 1N water An aqueous sodium oxide solution (2.0 ml) was added dropwise under ice cooling, and the mixture was stirred at room temperature for 2 hours. This solution was purified by dialysis 3 times against deionized water (total 20 hours, molecular weight cut off 6000 to 8000 Da), freeze-dried, and the title compound (207 mg, 0.0166 mmol, yield 73%) Was obtained as a white powder. 1 H-NMR (400 MHz, D 2 O) confirmed the completion of deprotection. The molecular weight distribution (Mw / Mn) was 1.06 by size exclusion chromatography.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 2.41-2.87 (m, 44H), 3.52-3, 78 (m, 909H), 4.30-4.53 (M, 16H).
合成例１３
ポリエチレングリコール（Ｍｗ＝１０，０００）－ベンジル保護ポリグルタミン酸（２５ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ（Ｂｎ）））の合成 Synthesis Example 13
Synthesis of polyethylene glycol (Mw = 10,000) -benzyl protected polyglutamic acid (25mer) -block copolymer (PEG-bP (Glu (Bn)))
Figure JPOXMLDOC01-appb-C000045
Figure JPOXMLDOC01-appb-C000045
　アルゴン雰囲気下にて、Ｈ２Ｎ－ＰＥＧ－ＯＭｅ（Ｍｗ＝１０，０００，３０４　ｍｇ，０．０３０４　ｍｍｏｌ）をジクロロメタン（１２．５　ｍｌ）に溶解させ、ＤＭＦ（３．０　ｍｌ）に溶解させたＢＬＧ－ＮＣＡ（２００　ｍｇ，０．７６０　ｍｍｏｌ）を滴下し、室温で１６時間攪拌した。氷冷下で反応液をヘキサン／酢酸エチル（３／２（ｖ／ｖ），５０　ｍｌ）中に滴下して再沈殿させ、標題化合物（３８４　ｍｇ，０．０２４８　ｍｍｏｌ，収率８２％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．３９、１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６）により、得られた化合物の重合度が２５であることを確認した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６）δ（ｐｐｍ）：１．８７－２．６８（ｍ，１６０Ｈ），３．４０－３．６２（ｍ，９０９Ｈ），３．７８－４．０７（ｍ，１８Ｈ），４．８７－５．１４（ｍ，４９Ｈ），７．１０－７．４６（ｍ，１２７Ｈ）． Under an argon atmosphere, H 2 N-PEG-OMe (Mw = 10,000, 304 mg, 0.0304 mmol) was dissolved in dichloromethane (12.5 ml) and dissolved in DMF (3.0 ml). BLG-NCA (200 mg, 0.760 mmol) was added dropwise and stirred at room temperature for 16 hours. The reaction mixture was re-precipitated dropwise in hexane / ethyl acetate (3/2 (v / v), 50 ml) under ice cooling to give the title compound (384 mg, 0.0248 mmol, yield 82%). Obtained as a white powder. It was confirmed by size exclusion chromatography that the molecular weight distribution (Mw / Mn) was 1.39 and the degree of polymerization of the obtained compound was 25 by 1 H-NMR (400 MHz, DMSO-d 6 ).
1 H-NMR (400 MHz, DMSO-d 6 ) δ (ppm): 1.87-2.68 (m, 160H), 3.40-3.62 (m, 909H), 3.78-4. 07 (m, 18H), 4.87-5.14 (m, 49H), 7.10-7.46 (m, 127H).
合成例１４
ポリエチレングリコール（Ｍｗ＝１０，０００）－ポリグルタミン酸（２５ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ））の合成 Synthesis Example 14
Synthesis of polyethylene glycol (Mw = 10,000) -polyglutamic acid (25mer) -block copolymer (PEG-bP (Glu))
Figure JPOXMLDOC01-appb-C000046
Figure JPOXMLDOC01-appb-C000046
　合成例１３で得られた化合物（２４８　ｍｇ，０．０１６０　ｍｍｏｌ）をＮＭＰ／Ｈ２Ｏ（１／１（ｖ／ｖ），１０　ｍｌ）に溶解させ、水酸化リチウム１水和物（６８．０　ｍｇ，１．６２　ｍｍｏｌ）を氷冷下で添加し、室温にて１．５時間攪拌した。この溶液を、脱イオン水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（１８５　ｍｇ，０．０１３９　ｍｍｏｌ，収率８７％）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）により、脱保護が完了したことを確認した。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．２７であった。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：１．７９－２．０９（ｍ，４７Ｈ），２．１１－２．３４（ｍ，４７Ｈ），３．４９－３．７３（ｍ，９０９Ｈ），４．１９－４．３２（ｍ，２１Ｈ）． The compound (248 mg, 0.0160 mmol) obtained in Synthesis Example 13 was dissolved in NMP / H 2 O (1/1 (v / v), 10 ml), and lithium hydroxide monohydrate (68. 0 mg, 1.62 mmol) was added under ice cooling, and the mixture was stirred at room temperature for 1.5 hours. This solution was purified by dialysis three times against deionized water (20 hours in total, molecular weight cut off 6000 to 8000 Da), lyophilized to give the title compound (185 mg, 0.0139 mmol, yield 87%). Was obtained as a white powder. 1 H-NMR (400 MHz, D 2 O) confirmed the completion of deprotection. The molecular weight distribution (Mw / Mn) was 1.27 by size exclusion chromatography.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 1.79-2.09 (m, 47H), 2.11-2.34 (m, 47H), 3.49-3.73 (M, 909H), 4.19-4.32 (m, 21H).
合成例１５
ポリエチレングリコール（Ｍｗ＝１０，０００）－ベンジル保護ポリグルタミン酸（５０ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ（Ｂｎ）））の合成 Synthesis Example 15
Synthesis of polyethylene glycol (Mw = 10,000) -benzyl protected polyglutamic acid (50mer) -block copolymer (PEG-bP (Glu (Bn)))
Figure JPOXMLDOC01-appb-C000047
Figure JPOXMLDOC01-appb-C000047
　アルゴン雰囲気下にて、Ｈ２Ｎ－ＰＥＧ－ＯＭｅ（Ｍｗ＝１０，０００，１．２９　ｇ，０．１２９　ｍｍｏｌ）、及びＢＬＧ－ＮＣＡ（１．６９　ｇ，６．４５　ｍｍｏｌ）より、合成例１３の方法と同様の方法で、標題化合物（９８８　ｍｇ，０．０４６９　ｍｍｏｌ，収率３７％）を得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．４７、１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６）により、得られた化合物の重合度が５０であることを確認した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６）δ（ｐｐｍ）：１．７６－２．７７（ｍ，２００Ｈ），３．４０－３．６０（ｍ，９０９Ｈ），３．７５－４．１４（ｍ，４３Ｈ），４．８７－５．１４（ｍ，９８Ｈ），７．１０－７．４６（ｍ，２５０Ｈ）． Synthesis example from H 2 N-PEG-OMe (Mw = 10,000, 1.29 g, 0.129 mmol) and BLG-NCA (1.69 g, 6.45 mmol) under argon atmosphere The title compound (988 mg, 0.0469 mmol, yield 37%) was obtained by a method similar to the method of 13. It was confirmed by size exclusion chromatography that the molecular weight distribution (Mw / Mn) was 1.47 and the degree of polymerization of the obtained compound was 50 by 1 H-NMR (400 MHz, DMSO-d 6 ).
1 H-NMR (400 MHz, DMSO-d 6 ) δ (ppm): 1.76-2.77 (m, 200H), 3.40-3.60 (m, 909H), 3.75-4. 14 (m, 43H), 4.87-5.14 (m, 98H), 7.10-7.46 (m, 250H).
合成例１６
ポリエチレングリコール（Ｍｗ＝１０，０００）－ポリグルタミン酸（５０ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ））の合成 Synthesis Example 16
Synthesis of polyethylene glycol (Mw = 10,000) -polyglutamic acid (50mer) -block copolymer (PEG-bP (Glu))
Figure JPOXMLDOC01-appb-C000048
Figure JPOXMLDOC01-appb-C000048
　合成例１５で得られた化合物（５００　ｍｇ，０．０２３７　ｍｍｏｌ）をＮＭＰ／Ｈ２Ｏ（１／１（ｖ／ｖ），１３．０　ｍｌ）に溶解させ、２Ｎ水酸化リチウム水溶液（２．６０　ｍｌ）を氷冷下で滴下し、室温にて２時間攪拌した。この溶液を、脱イオン水に対する３回の透析（計６時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（１９１　ｍｇ，０．０１１４　ｍｍｏｌ，収率４８％）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）により、脱保護が完了したことを確認した。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．３５であった。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：１．７５－２．０７（ｍ，１０２Ｈ），２．０９－２．３０（ｍ，９９Ｈ），３．４９－３．７３（ｍ，９０９Ｈ），４．１５－４．３４（ｍ，４５Ｈ）． The compound (500 mg, 0.0237 mmol) obtained in Synthesis Example 15 was dissolved in NMP / H 2 O (1/1 (v / v), 13.0 ml), and a 2N aqueous lithium hydroxide solution (2. 60 ml) was added dropwise under ice cooling, and the mixture was stirred at room temperature for 2 hours. This solution was purified by dialysis three times against deionized water (6 hours in total, fractional molecular weight 6000 to 8000 Da), lyophilized to give the title compound (191 mg, 0.0114 mmol, yield 48%). Was obtained as a white powder. 1 H-NMR (400 MHz, D 2 O) confirmed the completion of deprotection. The molecular weight distribution (Mw / Mn) was 1.35 by size exclusion chromatography.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 1.75-2.07 (m, 102H), 2.09-2.30 (m, 99H), 3.49-3.73 (M, 909H), 4.15-4.34 (m, 45H).
合成例１７
ポリエチレングリコール（Ｍｗ＝１０，０００）－ベンジル保護ポリグルタミン酸（２３ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ（Ｂｎ）））の合成 Synthesis Example 17
Synthesis of polyethylene glycol (Mw = 10,000) -benzyl protected polyglutamic acid (23mer) -block copolymer (PEG-bP (Glu (Bn)))
Figure JPOXMLDOC01-appb-C000049
Figure JPOXMLDOC01-appb-C000049
　アルゴン雰囲気下にて、Ｈ２Ｎ－ＰＥＧ－ＯＭｅ（Ｍｗ＝１０，０００，２．６０　ｇ，０．２６０　ｍｍｏｌ）をジクロロメタン（１２０　ｍｌ）に溶解させ、ＤＭＦ（３０　ｍｌ）に溶解させたＢＬＧ－ＮＣＡ（１．７０　ｇ，６．４６　ｍｍｏｌ）を滴下し、室温で１６時間攪拌した。氷冷下にて、反応液をヘキサン／酢酸エチル（３／２（ｖ／ｖ），７５０　ｍｌ）中に滴下して再沈殿させ、標題化合物（３．７０　ｇ，０．２４４　ｍｍｏｌ，収率９５％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．３９、１Ｈ－ＮＭＲ（ＤＭＳＯ－ｄ６）により、得られた化合物の重合度が２３であることを確認した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６）δ（ｐｐｍ）：１．５８－２．６４（ｍ，９２Ｈ），３．４３－３．６２（ｍ，９０９Ｈ），３．７７－４．１１（ｍ，１５Ｈ），４．８８－５．１４（ｍ，４５Ｈ），７．１２－７．４０（ｍ，１１５Ｈ）． Under an argon atmosphere, H 2 N-PEG-OMe (Mw = 10,000, 2.60 g, 0.260 mmol) was dissolved in dichloromethane (120 ml) and BLG dissolved in DMF (30 ml). -NCA (1.70 g, 6.46 mmol) was added dropwise and stirred at room temperature for 16 hours. Under ice-cooling, the reaction mixture was added dropwise to hexane / ethyl acetate (3/2 (v / v), 750 ml) for reprecipitation to give the title compound (3.70 g, 0.244 mmol, yield). 95%) was obtained as a white powder. It was confirmed by size exclusion chromatography that the molecular weight distribution (Mw / Mn) was 1.39 and the degree of polymerization of the obtained compound was 23 by 1 H-NMR (DMSO-d 6 ).
1 H-NMR (400 MHz, DMSO-d 6 ) δ (ppm): 1.58-2.64 (m, 92H), 3.43-3.62 (m, 909H), 3.77-4. 11 (m, 15H), 4.88-5.14 (m, 45H), 7.12-7.40 (m, 115H).
合成例１８
ポリエチレングリコール（Ｍｗ＝１０，０００）－ポリグルタミン酸（２３ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ））の合成 Synthesis Example 18
Synthesis of polyethylene glycol (Mw = 10,000) -polyglutamic acid (23mer) -block copolymer (PEG-bP (Glu))
Figure JPOXMLDOC01-appb-C000050
Figure JPOXMLDOC01-appb-C000050
　合成例１７で得られた化合物（１．７０　ｇ，０．１１２　ｍｍｏｌ）をＮＭＰ／Ｈ２Ｏ（１／１（ｖ／ｖ），１９．５　ｍｌ）に溶解させ、２Ｎ水酸化ナトリウム水溶液（６．５０　ｍｌ）を氷冷下で滴下し、室温にて２時間攪拌した。この溶液を、脱イオン水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（７８９　ｍｇ，０．０５８１　ｍｍｏｌ，収率５２％）を白色粉体として得た。１Ｈ－ＮＭＲ（Ｄ２Ｏ）により、脱保護が完了したことを確認した。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．１３であった。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：１．７９－２．０１（ｍ，４３Ｈ），２．１１－２．５４（ｍ，４２Ｈ），３．４９－３．７３（ｍ，９０９Ｈ），４．１９－４．３２（ｍ，２１Ｈ）． The compound obtained in Synthesis Example 17 (1.70 g, 0.112 mmol) was dissolved in NMP / H 2 O (1/1 (v / v), 19.5 ml), and 2N aqueous sodium hydroxide solution ( 6.50 ml) was added dropwise under ice cooling, and the mixture was stirred at room temperature for 2 hours. This solution was purified by dialysis three times against deionized water (20 hours in total, fractional molecular weight 6000 to 8000 Da), freeze-dried, and the title compound (789 mg, 0.0581 mmol, yield 52%) Was obtained as a white powder. 1 H-NMR (D 2 O) confirmed complete deprotection. The molecular weight distribution (Mw / Mn) was 1.13 by size exclusion chromatography.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 1.79-2.01 (m, 43H), 2.11-2.54 (m, 42H), 3.49-3.73 (M, 909H), 4.19-4.32 (m, 21H).
合成例１９
［Ｂ１２Ｈ１１－Ｃ４Ｈ８Ｏ２］テトラブチルアンモニウム塩の合成 Synthesis Example 19
Synthesis of [B 12 H 11 -C 4 H 8 O 2 ] tetrabutylammonium salt
Figure JPOXMLDOC01-appb-C000051
Figure JPOXMLDOC01-appb-C000051
　天然型Ｂ－ドデカハイドロドデカボレート・２テトラブチルアンモニウム塩（７．５０　ｇ，１２．０　ｍｍｏｌ）を１，４－ジオキサン（４００　ｍｌ）に溶解し、ＮａＢＦ４（６．５７　ｇ，５９．８　ｍｍｏｌ）、４Ｎ塩化水素の１，４－ジオキサン溶液（５．９９　ｍｌ，２３．９　ｍｍｏｌ）を加えた後、５時間加熱還流を行った。反応液を減圧濃縮後、イソプロパノールによってスラリー洗浄し、標題化合物（４．１７　ｇ，８．８５　ｍｍｏｌ，収率７４％）を白色粉体として得た。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤ３ＯＤ－ｄ４）δ（ｐｐｍ）：０．６３－１．９９（ｂｒ，１１Ｈ），１．０２（ｔ，１２Ｈ，Ｊ＝７．３　Ｈｚ），１．３９－１．４６（ｍ，８Ｈ），１．６２－１．７０（ｍ，８Ｈ），３．２１－３．２５（ｍ，８Ｈ），３．８６（ｔ，４Ｈ，Ｊ＝４．５　Ｈｚ），４．５４（ｔ，４Ｈ，Ｊ＝４．５　Ｈｚ）． Natural B-dodecahydrododecaborate-2 tetrabutylammonium salt (7.50 g, 12.0 mmol) was dissolved in 1,4-dioxane (400 ml), and NaBF 4 (6.57 g, 59.8) was dissolved. mmol), 1,4-dioxane solution (5.99 ml, 23.9 mmol) in 4N hydrogen chloride was added, followed by heating under reflux for 5 hours. The reaction mixture was concentrated under reduced pressure, and washed with slurry with isopropanol to give the title compound (4.17 g, 8.85 mmol, yield 74%) as a white powder.
1 H-NMR (400 MHz, CD 3 OD-d 4 ) δ (ppm): 0.63-1.99 (br, 11H), 1.02 (t, 12H, J = 7.3 Hz), 1 .39-1.46 (m, 8H), 1.62-1.70 (m, 8H), 3.21-3.25 (m, 8H), 3.86 (t, 4H, J = 4. 5 Hz), 4.54 (t, 4H, J = 4.5 Hz).
合成例２０
［Ｂ１２Ｈ１１－Ｏ（ＣＨ２）２Ｏ（ＣＨ２）２Ｎ３］２セシウム塩の合成 Synthesis Example 20
Synthesis of [B 12 H 11 —O (CH 2 ) 2 O (CH 2 ) 2 N 3 ] 2 cesium salt
Figure JPOXMLDOC01-appb-C000052
Figure JPOXMLDOC01-appb-C000052
　合成例１９で得られた化合物（４．１７　ｇ，８．８５　ｍｍｏｌ）を脱水ジクロロメタン（３０　ｍｌ）に溶解し、テトラブチルアンモニウムアジド（３．７７　ｇ，１３．３　ｍｍｏｌ）を加えた後、室温にて２時間攪拌した。反応液を減圧濃縮し、メタノール（５０　ｍｌ）に溶解させ、フッ化セシウム（２．６９　ｇ，１７．７　ｍｍｏｌ）を加えて１時間攪拌後、沈殿物をろ取した。ジクロロメタンによってスラリー洗浄し、標題化合物（４．４４　ｇ，８．２７　ｍｍｏｌ，収率９３％）を白色粉体として得た。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．２５－１．５４（ｂｒ，１１Ｈ），３．１９（ｔ，２Ｈ，Ｊ＝４．９　Ｈｚ），３．３３（ｍ，４Ｈ），３．３７－３．３９（ｍ，２Ｈ）． The compound obtained in Synthesis Example 19 (4.17 g, 8.85 mmol) was dissolved in dehydrated dichloromethane (30 ml), and tetrabutylammonium azide (3.77 g, 13.3 mmol) was added. Stir at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, dissolved in methanol (50 ml), cesium fluoride (2.69 g, 17.7 mmol) was added and stirred for 1 hour, and then the precipitate was collected by filtration. The slurry was washed with dichloromethane to give the title compound (4.44 g, 8.27 mmol, yield 93%) as a white powder.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.25 to 1.54 (br, 11H), 3.19 (t, 2H, J = 4.9 Hz), 3.33 ( m, 4H), 3.37-3.39 (m, 2H).
合成例２１
［Ｂ１２Ｈ１１－Ｏ（ＣＨ２）２Ｏ（ＣＨ２）２ＮＨ２］２テトラブチルアンモニウム塩の合成 Synthesis Example 21
Synthesis of [B 12 H 11 —O (CH 2 ) 2 O (CH 2 ) 2 NH 2 ] 2 tetrabutylammonium salt
Figure JPOXMLDOC01-appb-C000053
Figure JPOXMLDOC01-appb-C000053
　合成例２０で得られた化合物（２．２７　ｇ，４．２３　ｍｍｏｌ）を水（１００　ｍｌ）に溶解させ、テトラブチルアンモニウムブロミド（２．７３　ｇ，８．４６　ｍｍｏｌ）を加え、室温で１時間攪拌後、沈殿物をろ取し、［Ｂ１２Ｈ１１－Ｏ（ＣＨ２）２Ｏ（ＣＨ２）２Ｎ3］２テトラブチルアンモニウム塩（２．１７　ｇ，２．８７　ｍｍｏｌ，収率６８％）を得た。続いて、得られた化合物をメタノール（５７．５　ｍｌ）に溶解させ、アルゴン置換後、５％Ｐｄ／Ｃ（２．８７　ｇ，うち水分５２．５％）を加え、水素雰囲気下にて、５時間攪拌した。セライト（登録商標）濾過後、ろ液を減圧濃縮し、標題化合物（１．９１　ｇ，２．６３　ｍｍｏｌ，収率９１％）を白色粉体として得た。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤ３ＯＤ－ｄ４）δ（ｐｐｍ）：０．５８－１．８６（ｂｒ，１１Ｈ），１．０２（ｔ，１２Ｈ，Ｊ＝７．４　Ｈｚ），１．３７－１．４７（ｍ，８Ｈ），１．６２－１．７０（ｍ，８Ｈ），２．８３（ｔ，２Ｈ，Ｊ＝５．２　Ｈｚ），３．２１－３．２６（ｍ，８Ｈ），３．５５－３．６１（ｍ，４Ｈ），３．６９（ｔ，２Ｈ，Ｊ＝５．３　Ｈｚ）． The compound (2.27 g, 4.23 mmol) obtained in Synthesis Example 20 was dissolved in water (100 ml), tetrabutylammonium bromide (2.73 g, 8.46 mmol) was added, and 1 at room temperature was added. After stirring for a period of time, the precipitate was collected by filtration, and [B 12 H 11 —O (CH 2 ) 2 O (CH 2 ) 2 N 3 ] 2 tetrabutylammonium salt (2.17 g, 2.87 mmol, yield) 68%). Subsequently, the obtained compound was dissolved in methanol (57.5 ml), and after substitution with argon, 5% Pd / C (2.87 g, of which moisture was 52.5%) was added. Under a hydrogen atmosphere, Stir for 5 hours. After filtration through Celite (registered trademark), the filtrate was concentrated under reduced pressure to obtain the title compound (1.91 g, 2.63 mmol, yield 91%) as a white powder.
1 H-NMR (400 MHz, CD 3 OD-d 4 ) δ (ppm): 0.58-1.86 (br, 11H), 1.02 (t, 12H, J = 7.4 Hz), 1 .37-1.47 (m, 8H), 1.62-1.70 (m, 8H), 2.83 (t, 2H, J = 5.2 Hz), 3.21-3.26 (m , 8H), 3.55-3.61 (m, 4H), 3.69 (t, 2H, J = 5.3 Hz).
実施例４
ポリエチレングリコール（Ｍｗ＝１０，０００）－（ドデカボレート－アミド結合）ポリアスパラギン酸（２３ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ａｓｐ－Ｂ１２Ｈ１１））の合成 Example 4
Synthesis of polyethylene glycol (Mw = 10,000)-(dodecaborate-amide bond) polyaspartic acid (23mer) -block copolymer (PEG-bP (Asp-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000054
Figure JPOXMLDOC01-appb-C000054
　合成例１２で得られた化合物（５０．０　ｍｇ，３．８０　μｍｏｌ，Ａｓｐ残基：０．０８７４　ｍｏｌ）をＤＭＦ（２５．０　ｍｌ）に溶解し、４－（４，６－ジメトキシ－１，３，５－トリアジン－２－イル）－４－メチルモルホリニウムクロリド（ＤＭＴ－ＭＭ）（６４．０　ｍｇ，０．２３１　ｍｍｏｌ）、及びＮ－メチルモルホリン（ＮＭＭ）（２５．０　μｌ，０．２６９　ｍｍｏｌ）を加え、室温で２時間攪拌した。続いて、合成例２１で得られた化合物（１６８　ｍｇ，０．２３０　ｍｍｏｌ）を加えた後、室温で２０時間攪拌した。この溶液について、１５０　ｍＭ食塩水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）、及び脱イオン水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（４８．０　ｍｇ，３．５７　μｍｏｌ，収率８８％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．４２であった。ＩＣＰ－ＡＥＳ測定結果より、ポリアスパラギン酸（ＰＡｓｐ）の側鎖のカルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、２２％であった。これは、１ポリマーに対して、約５個の天然型Ｂ－ドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．０７－１．０９（ｍ，８６Ｈ），２．５０－３．０１（ｍ，３２Ｈ），３．１８－３．３５（ｍ，７Ｈ），３．３９－３．９５（ｍ，９０９Ｈ），４．４６－４．６０（ｍ，３Ｈ）． The compound obtained in Synthesis Example 12 (50.0 mg, 3.80 μmol, Asp residue: 0.0874 mol) was dissolved in DMF (25.0 ml) to give 4- (4,6-dimethoxy-1 , 3,5-triazin-2-yl) -4-methylmorpholinium chloride (DMT-MM) (64.0 mg, 0.231 mmol), and N-methylmorpholine (NMM) (25.0 μl, 0.269 mmol) was added, and the mixture was stirred at room temperature for 2 hours. Subsequently, the compound (168 mg, 0.230 mmol) obtained in Synthesis Example 21 was added, followed by stirring at room temperature for 20 hours. This solution was subjected to three dialysis against 150 mM saline (total 20 hours, fractional molecular weight 6000 to 8000 Da) and three times against deionized water (total 20 hours, fractional molecular weight 6000 to 8000 Da). Purification and lyophilization gave the title compound (48.0 mg, 3.57 μmol, 88% yield) as a white powder. The molecular weight distribution (Mw / Mn) was 1.42 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction rate of natural B-dodecaborate with respect to the carboxylic acid in the side chain of polyaspartic acid (PAsp) was 22%. This corresponds to the introduction of about 5 natural B-dodecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.07-1.09 (m, 86H), 2.50-3.01 (m, 32H), 3.18-3.35 (M, 7H), 3.39-3.95 (m, 909H), 4.46-4.60 (m, 3H).
実施例５
ポリエチレングリコール（Ｍｗ＝１０，０００）－（ドデカボレート－アミド結合）ポリグルタミン酸（２５ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｂ１２Ｈ１１））の合成 Example 5
Synthesis of polyethylene glycol (Mw = 10,000)-(dodecaborate-amide bond) polyglutamic acid (25mer) -block copolymer (PEG-bP (Glu-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000055
Figure JPOXMLDOC01-appb-C000055
　合成例１４で得られた化合物（２１．８　ｍｇ，１．６２　μｍｏｌ，Ｇｌｕ残基：０．０４０７　ｍｍｏｌ）をＤＭＳＯ（４．０　ｍｌ）に溶解し、Ｎ－ヒドロキシスクシンイミド（１９．５　ｍｇ，０．１６９　ｍｍｏｌ）、及び１－エチル－３－（３－ジメチルアミノプロピル）カルボジイミド塩酸塩（ＷＳＣ・ＨＣｌ）（３２．３　ｍｇ，０．１６９　ｍｍｏｌ）を加え、室温で２時間攪拌した。続いて、合成例２１で得られた化合物（１２３　ｍｇ，０．１６９　ｍｍｏｌ）、トリエチルアミン（４５．０　μｌ，０．３３９　ｍｍｏｌ）を加えた後、室温で２０時間攪拌した。この溶液について、１５０　ｍＭ食塩水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）、及び脱イオン水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（２２．０　ｍｇ，１．３１　μｍｏｌ，収率７９％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．５２であった。ＩＣＰ－ＡＥＳ測定結果より、ポリグルタミン酸（ＰＧｌｕ）の側鎖のカルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、５２％であった。これは、１ポリマーに対して、約１３個の天然型Ｂ－ドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．２１８－２．４８（ｍ，２８７Ｈ），２．２３－２．４８（ｍ，４７Ｈ），３．１８－３．４３（ｍ，４９Ｈ），３．６４－３．８７（ｍ，１２４９Ｈ），４．１６－４．４６（ｍ，１９Ｈ）． The compound obtained in Synthesis Example 14 (21.8 mg, 1.62 μmol, Glu residue: 0.0407 mmol) was dissolved in DMSO (4.0 ml), and N-hydroxysuccinimide (19.5 mg, 0.169 mmol) and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (WSC.HCl) (32.3 mg, 0.169 mmol) were added, and the mixture was stirred at room temperature for 2 hours. Subsequently, the compound obtained in Synthesis Example 21 (123 mg, 0.169 mmol) and triethylamine (45.0 μl, 0.339 mmol) were added, followed by stirring at room temperature for 20 hours. This solution was subjected to three dialysis against 150 mM saline (total 20 hours, fractional molecular weight 6000 to 8000 Da) and three times against deionized water (total 20 hours, fractional molecular weight 6000 to 8000 Da). Purification and lyophilization gave the title compound (22.0 mg, 1.31 μmol, yield 79%) as a white powder. The molecular weight distribution (Mw / Mn) was 1.52 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction rate of natural B-dodecaborate with respect to the carboxylic acid in the side chain of polyglutamic acid (PGlu) was 52%. This corresponds to the introduction of about 13 natural B-dodecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.218-2.48 (m, 287H), 2.23-2.48 (m, 47H), 3.18-3.43 (M, 49H), 3.64-3.87 (m, 1249H), 4.16-4.46 (m, 19H).
実施例６
ポリエチレングリコール（Ｍｗ＝１０，０００）－（ドデカボレート－アミド結合）ポリグルタミン酸（５０ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｂ１２Ｈ１１））の合成 Example 6
Synthesis of polyethylene glycol (Mw = 10,000)-(dodecaborate-amide bond) polyglutamic acid (50mer) -block copolymer (PEG-bP (Glu-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000056
Figure JPOXMLDOC01-appb-C000056
　合成例１６で得られた化合物（２０．０　ｍｇ，１．１９　μｍｏｌ，Ｇｌｕ残基：０．０５９７　ｍｍｏｌ）をＤＭＳＯ（４．０　ｍｌ）に溶解し、Ｎ－ヒドロキシスクシンイミド（３０．３　ｍｇ，０．２６３　ｍｍｏｌ）、及びＷＳＣ・ＨＣｌ（５０．４　ｍｇ，０．２６３　ｍｍｏｌ）を加え、室温で２時間攪拌した。続いて、合成例２１で得られた化合物（１９３　ｍｇ，０．２６３　ｍｍｏｌ）、トリエチルアミン（７０．０　μｌ，０．５２６　ｍｍｏｌ）を加えた後、室温で２０時間攪拌した。この溶液について、１５０　ｍＭ食塩水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）、及び脱イオン水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（１８．３　ｍｇ，０．７７６　μｍｏｌ，収率６５％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．３３であった。ＩＣＰ－ＡＥＳ測定結果より、ＰＧｌｕの側鎖のカルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、４８％であった。これは、１ポリマーに対して、約２４個の天然型Ｂ－ドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．１９－２．１９（ｍ，６３１Ｈ），２．２１－２．５３（ｍ，１００Ｈ），３．２５－３．３８（ｍ，１０３Ｈ），３．４０－３．８７（ｍ，１４７０Ｈ），４．１９－３．４２（ｍ，４５Ｈ）． The compound obtained in Synthesis Example 16 (20.0 mg, 1.19 μmol, Glu residue: 0.0597 mmol) was dissolved in DMSO (4.0 ml), and N-hydroxysuccinimide (30.3 mg, 0.263 mmol) and WSC · HCl (50.4 mg, 0.263 mmol) were added, and the mixture was stirred at room temperature for 2 hours. Subsequently, the compound obtained in Synthesis Example 21 (193 mg, 0.263 mmol) and triethylamine (70.0 μl, 0.526 mmol) were added, followed by stirring at room temperature for 20 hours. This solution was subjected to three dialysis against 150 mM saline (total 20 hours, fractional molecular weight 6000 to 8000 Da) and three times against deionized water (total 20 hours, fractional molecular weight 6000 to 8000 Da). Purification and lyophilization gave the title compound (18.3 mg, 0.776 μmol, 65% yield) as a white powder. The molecular weight distribution (Mw / Mn) was 1.33 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction rate of natural B-dodecaborate with respect to the carboxylic acid of the side chain of PGlu was 48%. This corresponds to the introduction of about 24 natural B-dodecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.19-2.19 (m, 631H), 2.21-2.53 (m, 100H), 3.25-3.38 (M, 103H), 3.40-3.87 (m, 1470H), 4.19-3.42 (m, 45H).
実施例７
ポリエチレングリコール（Ｍｗ＝１０，０００）－（ドデカボレート－アミド結合）ポリグルタミン酸（２３ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｂ１２Ｈ１１））の合成 Example 7
Synthesis of polyethylene glycol (Mw = 10,000)-(dodecaborate-amide bond) polyglutamic acid (23mer) -block copolymer (PEG-bP (Glu-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000057
Figure JPOXMLDOC01-appb-C000057
　合成例１８で得られた化合物（６００　ｍｇ，０．０４４５　ｍｍｏｌ，Ｇｌｕ残基：１．０２　ｍｍｏｌ）をＤＭＳＯ（１５　ｍｌ）に溶解し、Ｎ－ヒドロキシスクシンイミド（５８４　ｍｇ，５．１２　ｍｍｏｌ）、及びＷＳＣ・ＨＣｌ（９８２　ｍｇ，５．１２　ｍｍｏｌ）を加え、室温で２時間攪拌した。続いて、合成例２１で得られた化合物（２．２４　ｇ，４．１０　ｍｍｏｌ）、トリエチルアミン（１．３６　ｍｌ，１０．２　ｍｍｏｌ）を加えた後、室温で２０時間攪拌した。この溶液について、１５０　ｍＭ食塩水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）、及び脱イオン水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（７８０　ｍｇ，０．０４５９　ｍｍｏｌ，収率ｑｕａｎｔ．）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．２７であった。ＩＣＰ－ＡＥＳ測定結果より、ＰＧｌｕの側鎖のカルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、６１％であった。これは、１ポリマーに対して、約１４個の天然型Ｂ－ドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．０４－２．１５（ｍ，２１７Ｈ），２．１６－２．４３（ｍ，３８Ｈ），３．２１－３．４３（ｍ，４４Ｈ），３．５０－３．７０（ｍ，１０１４Ｈ），４．１９－４．４０（ｍ，２０Ｈ）． The compound obtained in Synthesis Example 18 (600 mg, 0.0445 mmol, Glu residue: 1.02 mmol) was dissolved in DMSO (15 ml), N-hydroxysuccinimide (584 mg, 5.12 mmol), And WSC · HCl (982 mg, 5.12 mmol) were added, and the mixture was stirred at room temperature for 2 hours. Subsequently, the compound obtained in Synthesis Example 21 (2.24 g, 4.10 mmol) and triethylamine (1.36 ml, 10.2 mmol) were added, followed by stirring at room temperature for 20 hours. This solution was subjected to three dialysis against 150 mM saline (total 20 hours, fractional molecular weight 6000 to 8000 Da) and three times against deionized water (total 20 hours, fractional molecular weight 6000 to 8000 Da). Purification and lyophilization gave the title compound (780 mg, 0.0459 mmol, yield quant.) As a white powder. The molecular weight distribution (Mw / Mn) was 1.27 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction ratio of natural B-dodecaborate to the carboxylic acid in the side chain of PGlu was 61%. This corresponds to the introduction of about 14 natural B-dodecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.04-2.15 (m, 217H), 2.16-2.43 (m, 38H), 3.21-3.43 (M, 44H), 3.50-3.70 (m, 1014H), 4.19-4.40 (m, 20H).
合成例２２
［Ｂ１２Ｈ１１－Ｏ（ＣＨ２）２Ｏ（ＣＨ２）２Ｓ（ＣＨ２）２ＮＨＢｏｃ］２－２ナトリウム塩の合成 Synthesis Example 22
[B 12 H 11 -O (CH 2) 2 O (CH 2) 2 S (CH 2) 2 NHBoc] 2- 2 Synthesis of sodium salt
Figure JPOXMLDOC01-appb-C000058
Figure JPOXMLDOC01-appb-C000058
　システアミン塩酸塩（５３０　ｍｇ，４．６７　ｍｍｏｌ）をジクロロメタン（１０　ｍｌ）に溶解させた後、ジクロロメタン（５．０　ｍｌ）に溶解させたＢｏｃ２Ｏ（１．４３　ｇ，６．５７　ｍｍｏｌ）を滴下した。続いてトリエチルアミン（７５４　μｌ，５．５９　ｍｍｏｌ）を加え、室温で２４時間攪拌した。飽和重層水、水、１５％食塩水にて順に洗浄し、無水硫酸マグネシウムで乾燥後、ろ過を行い、ろ液を減圧濃縮することで、保護システアミンの粗生成物（１．０３　ｇ）を得た。 Cysteamine hydrochloride (530 mg, 4.67 mmol) was dissolved in dichloromethane (10 ml), and then Boc 2 O (1.43 g, 6.57 mmol) dissolved in dichloromethane (5.0 ml) was added. It was dripped. Subsequently, triethylamine (754 μl, 5.59 mmol) was added, and the mixture was stirred at room temperature for 24 hours. Washing with saturated multistory water, water, and 15% saline in order, drying over anhydrous magnesium sulfate, filtering, and concentrating the filtrate under reduced pressure gave a crude product of protected cysteamine (1.03 g). It was.
　続いて得られた保護システアミンの粗生成物のうち、一部（２１０　ｍｇ）をアセトニトリル（５．０　ｍｌ）に溶解し、合成例１９で得られた化合物（２９７　ｍｇ，０．６３０　ｍｍｏｌ）と炭酸カリウム（２９９　ｍｇ，２．１６　ｍｍｏｌ）を加え、６０℃で２時間攪拌した。１Ｈ－ＮＭＲにて合成例１９で得られた化合物の残存が確認されたため、保護システアミンの粗生成物（２６８　ｍｇ）を加え、さらに１６時間攪拌した。反応液を濃縮後、シリカゲルカラムクロマトグラフィーにより精製し、標題化合物（２９５　ｍｇ，０．４３０　ｍｍｏｌ，収率６８％）を淡黄色粉体として得た。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤ３ＯＤ－ｄ４）δ（ｐｐｍ）：０．５８－１．８６（ｂｒ，１１Ｈ），１．０５（ｔ，１２Ｈ，Ｊ＝７．４　Ｈｚ），１．４０－１．４９（ｍ，８Ｈ），１．４６（ｓ，９Ｈ），１．６５－１．７３（ｍ，８Ｈ），２．６７（ｔ，２Ｈ，Ｊ＝７．１　Ｈｚ），２．７３（ｔ，　２Ｈ，Ｊ＝６．８　Ｈｚ），３．２３－３．２９（ｍ，８Ｈ），３．３２－３．３４（ｍ，２Ｈ），３．６１－３．７３（ｍ，６Ｈ）． Subsequently, a part (210 mg) of the obtained crude product of protected cysteamine was dissolved in acetonitrile (5.0 ml), and the compound (297 mg, 0.630 mmol) obtained in Synthesis Example 19 and Potassium carbonate (299 mg, 2.16 mmol) was added and stirred at 60 ° C. for 2 hours. Since the residue of the compound obtained in Synthesis Example 19 was confirmed by 1 H-NMR, a crude product of protected cysteamine (268 mg) was added, and the mixture was further stirred for 16 hours. The reaction mixture was concentrated and purified by silica gel column chromatography to give the title compound (295 mg, 0.430 mmol, yield 68%) as a pale-yellow powder.
1 H-NMR (400 MHz, CD 3 OD-d 4 ) δ (ppm): 0.58-1.86 (br, 11H), 1.05 (t, 12H, J = 7.4 Hz), 1 .40-1.49 (m, 8H), 1.46 (s, 9H), 1.65-1.73 (m, 8H), 2.67 (t, 2H, J = 7.1 Hz), 2.73 (t, 2H, J = 6.8 Hz), 3.23-3.29 (m, 8H), 3.32-3.34 (m, 2H), 3.61-3.73 ( m, 6H).
合成例２３
［Ｂ１２Ｈ１１－Ｏ（ＣＨ２）２Ｏ（ＣＨ２）２Ｓ（ＣＨ２）２ＮＨ２ＨＣｌ］２－の塩の合成 Synthesis Example 23
[B 12 H 11 —O (CH 2 ) 2 O (CH 2 ) 2 S (CH 2 ) 2 NH 2 HCl] Synthesis of 2-
Figure JPOXMLDOC01-appb-C000059
Figure JPOXMLDOC01-appb-C000059
　合成例２２で得られた化合物（２９５　ｍｇ，０．４３９　ｍｍｏｌ）をメタノール（１０　ｍｌ）に溶解し、４Ｎ塩酸の１，４－ジオキサン溶液（０．５０　ｍｌ，２．０　ｍｍｏｌ）を加え、１時間攪拌した。反応液を減圧濃縮することで、標題化合物（３０１　ｍｇ，０．４９６　ｍｍｏｌ，収率ｑｕａｎｔ．）を淡黄色粉体として得た。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤ３ＯＤ－ｄ４）δ（ｐｐｍ）：０．６３－１．８５（ｂｒ，１１Ｈ），１．０５（ｔ，１２Ｈ，Ｊ＝７．３　Ｈｚ），１．４０－１．４９（ｍ，８Ｈ），１．６５－１．７３（ｍ，８Ｈ），２．７８（ｔ，２Ｈ，Ｊ＝５．２　Ｈｚ），２．９７（ｔ，２Ｈ，Ｊ＝６．４　Ｈｚ），３．２２－３．２８（ｍ，８Ｈ），３．６１（ｔ，２Ｈ，Ｊ＝４．３　Ｈｚ），３．６８－３．７２（ｍ，４Ｈ），３．８２（ｔ，２Ｈ，Ｊ＝４．３　Ｈｚ）． The compound obtained in Synthesis Example 22 (295 mg, 0.439 mmol) was dissolved in methanol (10 ml), 4N hydrochloric acid in 1,4-dioxane (0.50 ml, 2.0 mmol) was added, Stir for 1 hour. The reaction mixture was concentrated under reduced pressure to give the title compound (301 mg, 0.496 mmol, yield quant.) As a pale yellow powder.
1 H-NMR (400 MHz, CD 3 OD-d 4 ) δ (ppm): 0.63-1.85 (br, 11H), 1.05 (t, 12H, J = 7.3 Hz), 1 40-1.49 (m, 8H), 1.65-1.73 (m, 8H), 2.78 (t, 2H, J = 5.2 Hz), 2.97 (t, 2H, J = 6.4 Hz), 3.22-3.28 (m, 8H), 3.61 (t, 2H, J = 4.3 Hz), 3.68-3.72 (m, 4H), 3 .82 (t, 2H, J = 4.3 Hz).
実施例８
ポリエチレングリコール（Ｍｗ＝１０，０００）－（ドデカボレート－アミド結合）ポリグルタミン酸（２５ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｂ１２Ｈ１１））の合成 Example 8
Synthesis of polyethylene glycol (Mw = 10,000)-(dodecaborate-amide bond) polyglutamic acid (25mer) -block copolymer (PEG-bP (Glu-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000060
Figure JPOXMLDOC01-appb-C000060
　合成例１４で得られた化合物（２０．０　ｍｇ，１．４９　μｍｏｌ，Ｇｌｕ残基：０．０３７３　ｍｍｏｌ）をＤＭＳＯ（４．０　ｍｌ）に溶解し、Ｎ－ヒドロキシスクシンイミド（１３．２　ｍｇ，０．１５５　ｍｍｏｌ）、及びＷＳＣ・ＨＣｌ（２９．７　ｍｇ，０．１５５　ｍｍｏｌ）を加え、室温で２時間攪拌した。続いて、合成例２３で得られた化合物（９４．２　ｍｇ，０．１５５　ｍｍｏｌ）、トリエチルアミン（３０．０　μｌ，０．２３０　ｍｍｏｌ）を加えた後、室温で２０時間攪拌した。この溶液について、１５０　ｍＭ食塩水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）、及び脱イオン水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（１８．０　ｍｇ，１．１３　μｍｏｌ，収率７３％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．３３であった。ＩＣＰ－ＡＥＳ測定結果より、ＰＧｌｕの側鎖カルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、３６％であった。これは、１ポリマーに対して、約９個の天然型Ｂ－ドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．１１９－２．１９（ｍ，３４１Ｈ），２．２０－２．４８（ｍ，５０Ｈ），２．５７－２．７９（ｍ，９３Ｈ），３．２６－３．４０（ｍ，７１Ｈ），３．４６－３．７６（ｍ，１７４４Ｈ），４．１１－３．４２（ｍ，２６Ｈ）． The compound obtained in Synthesis Example 14 (20.0 mg, 1.49 μmol, Glu residue: 0.0373 mmol) was dissolved in DMSO (4.0 ml), and N-hydroxysuccinimide (13.2 mg, 0.155 mmol) and WSC · HCl (29.7 mg, 0.155 mmol) were added, and the mixture was stirred at room temperature for 2 hours. Subsequently, the compound obtained in Synthesis Example 23 (94.2 mg, 0.155 mmol) and triethylamine (30.0 μl, 0.230 mmol) were added, followed by stirring at room temperature for 20 hours. This solution was subjected to three dialysis against 150 mM saline (total 20 hours, fractional molecular weight 6000 to 8000 Da) and three times against deionized water (total 20 hours, fractional molecular weight 6000 to 8000 Da). Purification and lyophilization gave the title compound (18.0 mg, 1.13 μmol, 73% yield) as a white powder. The molecular weight distribution (Mw / Mn) was 1.33 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction rate of natural B-dodecaborate with respect to the side chain carboxylic acid of PGlu was 36%. This corresponds to the introduction of about 9 natural B-dodecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.119-2.19 (m, 341H), 2.20-2.48 (m, 50H), 2.57-2.79 (M, 93H), 3.26-3.40 (m, 71H), 3.46-3.76 (m, 1744H), 4.11-3.42 (m, 26H).
合成例２４
［Ｂ１２Ｈ１１－Ｏ（ＣＨ２）２Ｏ（ＣＨ２）２ＳＣ（ＮＨ２）２］－テトラブチルアンモニウム塩の合成 Synthesis Example 24
[B 12 H 11 -O (CH 2) 2 O (CH 2) 2 SC (NH 2) 2] - Synthesis of tetrabutylammonium salt
Figure JPOXMLDOC01-appb-C000061
Figure JPOXMLDOC01-appb-C000061
　合成例１９で得られた化合物（６００　ｍｇ，１．２７　ｍｍｏｌ）をエタノール（３０　ｍｌ）に溶解させ、チオ尿素（１９４　ｍｇ，２．５６　ｍｍｏｌ）を加え、８５℃で１６時間攪拌した。室温に戻し、生じた沈殿をろ取して、標題化合物（３５０　ｍｇ，０．６３９　ｍｍｏｌ，収率５０％）を白色粉体として得た。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６）δ（ｐｐｍ）：０．２５－１．７６（ｂｒ，１１Ｈ），０．９２（ｔ，１２Ｈ，Ｊ＝７．３　Ｈｚ），１．２７－１．３６（ｍ，８Ｈ），１．５３－１．６１（ｍ，８Ｈ），３．３０－３．３８（ｍ，２Ｈ），３．４３－３．４６（ｍ，２Ｈ），３．４９－３．５２（ｍ，２Ｈ），３．６８（ｔ，２Ｈ，Ｊ＝５．５　Ｈｚ）． The compound (600 mg, 1.27 mmol) obtained in Synthesis Example 19 was dissolved in ethanol (30 ml), thiourea (194 mg, 2.56 mmol) was added, and the mixture was stirred at 85 ° C. for 16 hours. The temperature was returned to room temperature, and the resulting precipitate was collected by filtration to give the title compound (350 mg, 0.639 mmol, yield 50%) as a white powder.
1 H-NMR (400 MHz, DMSO-d 6 ) δ (ppm): 0.25-1.76 (br, 11H), 0.92 (t, 12H, J = 7.3 Hz), 1.27 -1.36 (m, 8H), 1.53-1.61 (m, 8H), 3.30-3.38 (m, 2H), 3.43-3.46 (m, 2H), 3 .49-3.52 (m, 2H), 3.68 (t, 2H, J = 5.5 Hz).
合成例２５
［Ｂ１２Ｈ１１－Ｏ（ＣＨ２）２Ｏ（ＣＨ２）２ＳＨ］２－２テトラブチルアンモニウム塩の合成 Synthesis Example 25
[B 12 H 11 -O (CH 2) 2 O (CH 2) 2 SH] 2- 2 Synthesis of tetrabutylammonium salt
Figure JPOXMLDOC01-appb-C000062
Figure JPOXMLDOC01-appb-C000062
　合成例２４で得られた化合物（３５０　ｍｇ，０．６３９　ｍｍｏｌ）をメタノール（３０　ｍｌ）に溶解させ、５Ｍナトリウムメトキシド／メタノール（０．５１０　ｍｌ，２．５６　ｍｍｏｌ）を加え、室温で２時間半攪拌した。その後、反応液を減圧濃縮し、水（１０　ｍｌ）に再度溶解させ、酢酸を添加してｐＨを４．０に合わせた。続いて、水（５．０　ｍｌ）に溶解させたテトラブチルアンモニウムブロミド（４１２　ｍｇ，１．２８　ｍｍｏｌ）を加え、生じた沈殿物をろ取し、標題化合物（３８９　ｍｇ，０．５２４　ｍｍｏｌ，収率８２％）を白色粉体として得た。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤ３ＯＤ－ｄ４）δ（ｐｐｍ）：０．４９－１．７６（ｂｒ，１１Ｈ），０．９２（ｔ，１２Ｈ，Ｊ＝７．３　Ｈｚ），１．２９－１．３８（ｍ，８Ｈ），１．５４－１．６２（ｍ，８Ｈ），２．５３（ｔ，２Ｈ，Ｊ＝６．６　Ｈｚ），３．１６－３．１８（ｍ，８Ｈ），３．４９－３．５３（ｍ，４Ｈ），３．５９－３．６４（ｍ，２Ｈ）． The compound (350 mg, 0.639 mmol) obtained in Synthesis Example 24 was dissolved in methanol (30 ml), 5M sodium methoxide / methanol (0.510 ml, 2.56 mmol) was added, and 2 at room temperature was added. Stir for half an hour. Thereafter, the reaction solution was concentrated under reduced pressure, redissolved in water (10 ml), and acetic acid was added to adjust the pH to 4.0. Subsequently, tetrabutylammonium bromide (412 mg, 1.28 mmol) dissolved in water (5.0 ml) was added, and the resulting precipitate was collected by filtration to give the title compound (389 mg, 0.524 mmol, Yield 82%) was obtained as a white powder.
1 H-NMR (400 MHz, CD 3 OD-d 4 ) δ (ppm): 0.49-1.76 (br, 11H), 0.92 (t, 12H, J = 7.3 Hz), 1 29-1.38 (m, 8H), 1.54-1.62 (m, 8H), 2.53 (t, 2H, J = 6.6 Hz), 3.16-3.18 (m , 8H), 3.49-3.53 (m, 4H), 3.59-3.64 (m, 2H).
実施例９
ポリエチレングリコール（Ｍｗ＝１０，０００）－（ドデカボレート－チオエステル結合）ポリグルタミン酸（２３ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｂ１２Ｈ１１））の合成 Example 9
Synthesis of polyethylene glycol (Mw = 10,000)-(dodecaborate-thioester bond) polyglutamic acid (23mer) -block copolymer (PEG-bP (Glu-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000063
Figure JPOXMLDOC01-appb-C000063
　合成例１８で得られた化合物（１０．０　ｍｇ，０．７４２　μｍｏｌ，Ｇｌｕ残基：０．０１６９　μｍｏｌ）をＮ－メチル－２－ピロリドン（ＮＭＰ）（５．０　ｍｌ）に溶解し、Ｎ－ヒドロキシスクシンイミド（９．７０　ｍｇ，０．０８４７　μｍｏｌ）、及びＷＳＣ・ＨＣｌ（１６．２　ｍｇ，０．０８４７　ｍｍｏｌ）を加え、室温で２時間攪拌した。続いて、合成例２５で得られた化合物（６３．３　ｍｇ，０．８４７　ｍｍｏｌ）、及びＮ，Ｎ－ジメチル－４－アミノピリジン（ＤＭＡＰ）（２．１　ｍｇ，０．０１６９　ｍｍｏｌ）を加えた後、室温で１６時間攪拌した。この溶液について、１５０　ｍＭ食塩水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）、及び脱イオン水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（９．８０　ｍｇ，０．０５６０　ｍｍｏｌ，収率７５％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．１２であった。ＩＣＰ－ＡＥＳ測定結果より、ＰＧｌｕの側鎖のカルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、６５％であった。これは、１ポリマーに対して、約１５個の天然型Ｂ－ドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．３２－２．２１（ｍ，１２２Ｈ），２．１９－２．７０（ｍ，４２Ｈ），３．５１－３．８４（ｍ，９９９Ｈ），４．１１－４．４２（ｍ，６Ｈ）． The compound obtained in Synthesis Example 18 (10.0 mg, 0.742 μmol, Glu residue: 0.0169 μmol) was dissolved in N-methyl-2-pyrrolidone (NMP) (5.0 ml). -Hydroxysuccinimide (9.70 mg, 0.0847 μmol) and WSC · HCl (16.2 mg, 0.0847 mmol) were added, and the mixture was stirred at room temperature for 2 hours. Subsequently, the compound obtained in Synthesis Example 25 (63.3 mg, 0.847 mmol) and N, N-dimethyl-4-aminopyridine (DMAP) (2.1 mg, 0.0169 mmol) were added. Then, the mixture was stirred at room temperature for 16 hours. This solution was subjected to three dialysis against 150 mM saline (total 20 hours, fractional molecular weight 6000 to 8000 Da) and three times against deionized water (total 20 hours, fractional molecular weight 6000 to 8000 Da). Purification and lyophilization gave the title compound (9.80 mg, 0.0560 mmol, 75% yield) as a white powder. The molecular weight distribution (Mw / Mn) was 1.12 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction rate of natural B-dodecaborate with respect to the carboxylic acid of the side chain of PGlu was 65%. This corresponds to the introduction of about 15 natural B-dodecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.32-2.21 (m, 122H), 2.19-2.70 (m, 42H), 3.51-3.84 (M, 999H), 4.11-4.42 (m, 6H).
合成例２６
ポリエチレングリコール（Ｍｗ＝１０，０００）－末端アルキン結合ポリグルタミン酸（２３ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｂ１２Ｈ１１））の合成 Synthesis Example 26
Synthesis of polyethylene glycol (Mw = 10,000) -terminal alkyne-linked polyglutamic acid (23mer) -block copolymer (PEG-bP (Glu-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000064
Figure JPOXMLDOC01-appb-C000064
　合成例１８で得られた化合物（１５０　ｍｇ，１１．１　μｍｏｌ，Ｇｌｕ残基：０．２５４　ｍｍｏｌ）をＮＭＰ（１０　ｍｌ）に溶解し、Ｎ－ヒドロキシスクシンイミド（１４６　ｍｇ，１．２７　ｍｍｏｌ）、及びＷＳＣ・ＨＣｌ（２４３　ｍｇ，１．２７　ｍｍｏｌ）を加え、室温で２時間攪拌した。続いて、２－［２－（２－プロピニルオキシ）エトキシ］エチルアミン（１８２　ｍｇ，１．２７　ｍｍｏｌ）、及びトリエチルアミン（３５４　μｌ，２．５４　ｍｍｏｌ）を加えた後、室温で１６時間攪拌した。この溶液について、１５０　ｍＭ食塩水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）、及び脱イオン水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（１５４　ｍｇ，１０．２　μｍｏｌ，収率９２％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．０７であった。１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）測定結果より、ＰＧｌｕの側鎖のカルボン酸に対する２－［２－（２－プロピニルオキシ）エトキシ］エチルアミンの導入率は７０％であった。これは、１ポリマーに対して、約１６個の２－［２－（２－プロピニルオキシ）エトキシ］エチルアミンが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：１．７８－２．１３（ｍ，４１Ｈ），２．１６－２．４０（ｍ，４１Ｈ），２．７７－２．８３（ｍ，１４Ｈ），３．２２－３．３８（ｍ，３２Ｈ），３．５６－３．７３（ｍ，１１７２Ｈ），４．１０－４．３１（ｍ，１７Ｈ）． The compound obtained in Synthesis Example 18 (150 mg, 11.1 μmol, Glu residue: 0.254 mmol) was dissolved in NMP (10 ml), N-hydroxysuccinimide (146 mg, 1.27 mmol), And WSC · HCl (243 mg, 1.27 mmol) were added, and the mixture was stirred at room temperature for 2 hours. Subsequently, 2- [2- (2-propynyloxy) ethoxy] ethylamine (182 mg, 1.27 mmol) and triethylamine (354 μl, 2.54 mmol) were added, followed by stirring at room temperature for 16 hours. This solution was subjected to three dialysis against 150 mM saline (total 20 hours, fractional molecular weight 6000 to 8000 Da) and three times against deionized water (total 20 hours, fractional molecular weight 6000 to 8000 Da). Purification and lyophilization gave the title compound (154 mg, 10.2 μmol, 92% yield) as a white powder. The molecular weight distribution (Mw / Mn) was 1.07 by size exclusion chromatography. From the result of 1 H-NMR (400 MHz, D 2 O) measurement, the introduction rate of 2- [2- (2-propynyloxy) ethoxy] ethylamine with respect to the carboxylic acid in the side chain of PGlu was 70%. This corresponds to the introduction of about 16 2- [2- (2-propynyloxy) ethoxy] ethylamine per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 1.78-2.13 (m, 41H), 2.16-2.40 (m, 41H), 2.77-2.83 (M, 14H), 3.22-3.38 (m, 32H), 3.56-3.73 (m, 1172H), 4.10-4.31 (m, 17H).
実施例１０
ポリエチレングリコール（Ｍｗ＝１０，０００）－（ドデカボレート－トリアゾール結合）ポリグルタミン酸（２３ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｂ１２Ｈ１１））の合成 Example 10
Synthesis of polyethylene glycol (Mw = 10,000)-(dodecaborate-triazole bond) polyglutamic acid (23mer) -block copolymer (PEG-bP (Glu-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000065
Figure JPOXMLDOC01-appb-C000065
　合成例２６で得られた化合物（１０．０　ｍｇ，０．６６１　μｍｏｌ，Ｇｌｕ残基：１５．２　μｍｏｌ）を１００　ｍＭリン酸緩衝液（１０　ｍｌ）に溶解させ、合成例２０で得られた化合物（１６．３　ｍｇ，３０．４　μｍｏｌ）、Ｎ，Ｎ，Ｎ’，Ｎ’’，Ｎ’’－ペンタメチルジエチレントリアミン（ＰＭＤＥＴＡ）（５．０　μｌ，２８．９　μｍｏｌ）、及びトリス（３－ヒドロキシプロピルトリアゾリルメチル）アミン（ＴＨＰＴＡ）（１９．８　ｍｇ，４５．５　μｍｏｌ）を加え、アルゴンガスで５分間バブリングした。続いて、臭化銅（Ｉ）（１．３０　ｍｇ，９．０６　μｍｏｌ）を加え、アルゴン雰囲気下で室温にて９６時間攪拌した。この溶液について、１５０　ｍＭ食塩水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）、及び脱イオン水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物を（１３．８　ｍｇ，０．６９８　μｍｏｌ，収率ｑｕａｎｔ．）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．２４であった。ＩＣＰ－ＡＥＳ測定結果より、ＰＧｌｕの側鎖のカルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、３９％であった。これは、１ポリマーに対して、約９個の天然型Ｂ－ドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．１９９－２．１５（ｍ，１８７Ｈ），２．１５－２．５６（ｍ，４６Ｈ），３．１４－３．３９（ｍ，１８Ｈ），３．３９－３．７１（ｍ，１３９８Ｈ），４．０６－４．３６（ｍ，２７Ｈ）． The compound obtained in Synthesis Example 26 (10.0 mg, 0.661 μmol, Glu residue: 15.2 μmol) was dissolved in 100 mM phosphate buffer (10 ml) and obtained in Synthesis Example 20. Compound (16.3 mg, 30.4 μmol), N, N, N ′, N ″, N ″ -pentamethyldiethylenetriamine (PMDETA) (5.0 μl, 28.9 μmol), and Tris (3 -Hydroxypropyltriazolylmethyl) amine (THPTA) (19.8 mg, 45.5 μmol) was added and bubbled with argon gas for 5 minutes. Subsequently, copper (I) bromide (1.30 mg, 9.06 μmol) was added, and the mixture was stirred at room temperature for 96 hours under an argon atmosphere. This solution was subjected to three dialysis against 150 mM saline (total 20 hours, fractional molecular weight 6000 to 8000 Da) and three times against deionized water (total 20 hours, fractional molecular weight 6000 to 8000 Da). Purification and lyophilization gave the title compound (13.8 mg, 0.698 μmol, yield quant.) As a white powder. The molecular weight distribution (Mw / Mn) was 1.24 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction rate of natural B-dodecaborate with respect to carboxylic acid in the side chain of PGlu was 39%. This corresponds to the introduction of about 9 natural B-dodecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.199-2.15 (m, 187H), 2.15-2.56 (m, 46H), 3.14-3.39 (M, 18H), 3.39-3.71 (m, 1398H), 4.06-4.36 (m, 27H).
合成例２７
Ｎ－保護アミノヘキサン酸活性エステルの合成 Synthesis Example 27
Synthesis of N-protected aminohexanoic acid active ester
Figure JPOXMLDOC01-appb-C000066
Figure JPOXMLDOC01-appb-C000066
　６-アミノヘキサン酸（８４８　ｍｇ，６．４８　ｍｍｏｌ）を水（１０　ｍｌ）に溶解させた後、ＮａＨＣＯ３（８１２　ｍｇ，９．６７　ｍｍｏｌ）を添加した。続いてＣｂｚ－Ｃｌ（１．１　ｍｌ，７．７８　ｍｍｏｌ）を加え、室温で２４時間攪拌した。水層を、濃塩酸を用いてｐＨ１．０にし、ジクロロメタンで抽出後、１５％食塩水で洗浄した。有機層を無水硫酸マグネシウムで乾燥後、ろ過を行い、ろ液を減圧濃縮することで、Ｎ－保護アミノヘキサン酸（１．２　ｇ，４．５４　ｍｍｏｌ，収率７０％）を白色紛体として得た。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤ３ＯＤ－ｄ４）δ（ｐｐｍ）：１．３１－１．４２（ｍ，２Ｈ），１．４９－１．５６（ｍ，２Ｈ），１．５９－１．６８（ｍ，２Ｈ），２．３０（ｔ，２Ｈ，Ｊ＝７．４　Ｈｚ），３．１４（ｑ，２Ｈ，Ｊ＝６．８　Ｈｚ，１３．２　Ｈｚ），５．０８（ｓ，２Ｈ），７．２４－７．３６（ｍ，５Ｈ）． After dissolving 6-aminohexanoic acid (848 mg, 6.48 mmol) in water (10 ml), NaHCO 3 (812 mg, 9.67 mmol) was added. Subsequently, Cbz-Cl (1.1 ml, 7.78 mmol) was added, and the mixture was stirred at room temperature for 24 hours. The aqueous layer was adjusted to pH 1.0 using concentrated hydrochloric acid, extracted with dichloromethane, and washed with 15% brine. The organic layer is dried over anhydrous magnesium sulfate and filtered, and the filtrate is concentrated under reduced pressure to obtain N-protected aminohexanoic acid (1.2 g, 4.54 mmol, yield 70%) as a white powder. It was.
1 H-NMR (400 MHz, CD 3 OD-d 4 ) δ (ppm): 1.31-1.42 (m, 2H), 1.49-1.56 (m, 2H), 1.59- 1.68 (m, 2H), 2.30 (t, 2H, J = 7.4 Hz), 3.14 (q, 2H, J = 6.8 Hz, 13.2 Hz), 5.08 ( s, 2H), 7.24-7.36 (m, 5H).
　得られたＮ－保護アミノヘキサン酸（１．２　ｇ，４．５４　ｍｍｏｌ）をアセトニトリル（１０　ｍｌ）に溶解し、クロロぎ酸４-ニトロフェニル（１．４７　ｍｌ，７．２９　ｍｍｏｌ）、トリエチルアミン（０．９４４　ｍｌ，７．０９　ｍｍｏｌ）、ＤＭＡＰ（８２．０　ｍｇ，０．６６８　ｍｍｏｌ）を加え、４℃で４５分攪拌した。反応液を濃縮後、シリカゲルカラムクロマトグラフィーにより精製し、標題化合物（６３２　ｍｇ，１．６４　ｍｍｏｌ，収率３６％）を淡黄色油状物質として得た。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤ３ＯＤ－ｄ４）δ（ｐｐｍ）：１．４４－１．５１（ｍ，２Ｈ），１．５５－１．６２（ｍ，２Ｈ），１．７５－１．８２（ｍ，２Ｈ），２．６６（ｔ，２Ｈ，Ｊ＝７．４　Ｈｚ），３．１７（ｔ，２Ｈ，Ｊ＝６．８　Ｈｚ），５．０８（ｓ，２Ｈ），７．２９－７．３８（ｍ，７Ｈ），８．３０（ｄ，２Ｈ，Ｊ＝９．２　Ｈｚ）． The obtained N-protected aminohexanoic acid (1.2 g, 4.54 mmol) was dissolved in acetonitrile (10 ml), 4-nitrophenyl chloroformate (1.47 ml, 7.29 mmol), triethylamine. (0.944 ml, 7.09 mmol) and DMAP (82.0 mg, 0.668 mmol) were added and stirred at 4 ° C. for 45 minutes. The reaction mixture was concentrated and purified by silica gel column chromatography to give the title compound (632 mg, 1.64 mmol, yield 36%) as a pale yellow oil.
1 H-NMR (400 MHz, CD 3 OD-d 4 ) δ (ppm): 1.44 to 1.51 (m, 2H), 1.55 to 1.62 (m, 2H), 1.75- 1.82 (m, 2H), 2.66 (t, 2H, J = 7.4 Hz), 3.17 (t, 2H, J = 6.8 Hz), 5.08 (s, 2H), 7.29-7.38 (m, 7H), 8.30 (d, 2H, J = 9.2 Hz).
合成例２８
アミノドデカボレートの合成 Synthesis Example 28
Synthesis of aminododecaborate
Figure JPOXMLDOC01-appb-C000067
Figure JPOXMLDOC01-appb-C000067
　天然型Ｂ－ドデカハイドロドデカボレート・２ナトリウム塩（３．０　ｇ，１５．９　ｍｍｏｌ）を水（５０　ｍｌ）に溶解させた後、ヒドロキシルアミン-Ｏ-スルホン酸（３．６　ｇ，３１．８　ｍｍｏｌ）を加え、１１０℃で２８時間攪拌した。続いて氷浴中でテトラメチルアンモニウムクロリド（５．２　ｇ,　４７．９　ｍｍｏｌ）を添加し攪拌した。析出物をろ取した。この析出物を水に溶解後、４℃に冷却して晶析させ、ろ取し、真空乾燥することで標題化合物（１．２　ｇ，収率２０％）を白色粉体として得た。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６）δ（ｐｐｍ）：０．１５６－１．９４（ｍ，１１Ｈ），３．０９（ｓ，１２Ｈ），５．３５－５．７６（ｂｒ，３Ｈ）. Natural B-dodecahydrododecaborate disodium salt (3.0 g, 15.9 mmol) was dissolved in water (50 ml), and then hydroxylamine-O-sulfonic acid (3.6 g, 31.31 g). 8 mmol) was added, and the mixture was stirred at 110 ° C. for 28 hours. Subsequently, tetramethylammonium chloride (5.2 g, 47.9 mmol) was added and stirred in an ice bath. The precipitate was collected by filtration. The precipitate was dissolved in water, crystallized by cooling to 4 ° C., collected by filtration and dried in vacuo to give the title compound (1.2 g, yield 20%) as a white powder.
1 H-NMR (400 MHz, DMSO-d 6 ) δ (ppm): 0.156-1.94 (m, 11H), 3.09 (s, 12H), 5.35-5.76 (br, 3H).
合成例２９
アミノヘキサン酸―ドデカボレートアミドの合成 Synthesis Example 29
Synthesis of aminohexanoic acid-dodecaborate amide
Figure JPOXMLDOC01-appb-C000068
Figure JPOXMLDOC01-appb-C000068
　合成例２８で得られた化合物（２３０　ｍｇ，０．６４５　μｍｏｌ）をＤＭＦ（５．０　ｍｌ）に溶解し、ＮａＨ（６３．０　ｍｇ，２．５８　ｍｍｏｌ）を加え、５０℃で１時間攪拌した。続いて、合成例２７で得られた化合物（２９９　ｍｇ，０．７７４　ｍｍｏｌ）を加え、５０℃で１６時間攪拌した。反応液を濃縮後、ジクロロメタン（５　ｍｌ）を加え、不溶物をろ去した。続いてろ液にヘキサン（２５　ｍｌ）を加えて沈殿を生じさせ、上清をデカンテーションにより除去し、Ｎ－保護アミノヘキサン酸―ドデカボレートアミド（１５８　ｍｇ，０．３２９　μｍｏｌ，収率４３％）を淡黄色粘性物質として得た。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＤＭＳＯ－ｄ６）δ（ｐｐｍ）：０．４５－２．１７（ｍ，１１Ｈ）,１．３３－１．４６（ｍ，２Ｈ）,１．４７－１．７４（ｍ，４Ｈ）,２．
５９－２．７５（ｍ，２Ｈ）,３．０９－３．１７（ｍ，２Ｈ）,５．０４－５．１０（ｓ，２Ｈ）,７．２５－７．４２（ｍ，５Ｈ）. The compound (230 mg, 0.645 μmol) obtained in Synthesis Example 28 was dissolved in DMF (5.0 ml), NaH (63.0 mg, 2.58 mmol) was added, and the mixture was stirred at 50 ° C. for 1 hour. did. Subsequently, the compound obtained in Synthesis Example 27 (299 mg, 0.774 mmol) was added, and the mixture was stirred at 50 ° C. for 16 hours. The reaction mixture was concentrated, dichloromethane (5 ml) was added, and insoluble material was removed by filtration. Subsequently, hexane (25 ml) was added to the filtrate to cause precipitation, the supernatant was removed by decantation, and N-protected aminohexanoic acid-dodecaborateamide (158 mg, 0.329 μmol, yield 43%) Was obtained as a pale yellow viscous substance.
1 H-NMR (400 MHz, DMSO-d 6 ) δ (ppm): 0.45-2.17 (m, 11H), 1.33-1.46 (m, 2H), 1.47-1. 74 (m, 4H), 2.
59-2.75 (m, 2H), 3.09-3.17 (m, 2H), 5.04-5.10 (s, 2H), 7.25-7.42 (m, 5H).
　得られた化合物の一部（５６．０　ｍｇ，０．１１７　ｍｍｏｌ）をメタノール（８．０　ｍｌ）に溶解させ、アルゴン雰囲気下で、５％Ｐｄ／Ｃ（６０．０　ｍｇ，うち水分５２．５％）を加えた。続いて水素置換し、水素雰囲気下で５時間攪拌した。ろ過を行い、ろ液を減圧濃縮して標題化合物（３１．０　ｍｇ，０．０９７４　ｍｍｏｌ，収率８３％）を褐色固体として得た。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤ３ＯＤ－ｄ４）δ（ｐｐｍ）：０．５８－２．０３（ｂｒ，１１Ｈ），１．３５－１．４４（ｍ，２Ｈ），１．４７－１．５６（ｍ，２Ｈ），１．５８－１．６６（ｍ，２Ｈ），２．６６（ｔ，２Ｈ，Ｊ＝７．９　Ｈｚ），３．１４（ｔ，２Ｈ，Ｊ＝６．９　Ｈｚ），５．０８（ｓ，２Ｈ），７．２９－７．３９（ｍ，５Ｈ）． Part of the obtained compound (56.0 mg, 0.117 mmol) was dissolved in methanol (8.0 ml), and 5% Pd / C (60.0 mg, of which water content was 52. 5 mg) under an argon atmosphere. 5%) was added. Subsequently, the atmosphere was replaced with hydrogen, and the mixture was stirred for 5 hours under a hydrogen atmosphere. Filtration was performed, and the filtrate was concentrated under reduced pressure to obtain the title compound (31.0 mg, 0.0974 mmol, yield 83%) as a brown solid.
1 H-NMR (400 MHz, CD 3 OD-d 4 ) δ (ppm): 0.58 to 2.03 (br, 11H), 1.35 to 1.44 (m, 2H), 1.47— 1.56 (m, 2H), 1.58-1.66 (m, 2H), 2.66 (t, 2H, J = 7.9 Hz), 3.14 (t, 2H, J = 6. 9 Hz), 5.08 (s, 2H), 7.29-7.39 (m, 5H).
実施例１１
ポリエチレングリコール（Ｍｗ＝１０，０００）－（ドデカボレート－アミド結合）ポリグルタミン酸（２３ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－ＨＮ－Ｂ１２Ｈ１１））の合成 Example 11
Synthesis of polyethylene glycol (Mw = 10,000)-(dodecaborate-amide bond) polyglutamic acid (23mer) -block copolymer (PEG-bP (Glu-HN-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000069
Figure JPOXMLDOC01-appb-C000069
　合成例１８で得られた化合物（１０　ｍｇ，０．７３４　μｍｏｌ，Ｇｌｕ残基：０．０１６９　ｍｍｏｌ）をＤＭＳＯ（５．０　ｍｌ）に溶解し、Ｎ－ヒドロキシスクシンイミド（９．７　ｍｇ，０．０８４５　ｍｍｏｌ）、及びＷＳＣ・ＨＣｌ（１６．０　ｍｇ，０．０８４５　ｍｍｏｌ）を加え、室温で２時間攪拌した。続いて、合成例２９で得られた化合物（２９　ｍｇ，０．８４７　ｍｍｏｌ）、トリエチルアミン（２３．６　μｌ，０．１６９　ｍｍｏｌ）を加えた後、室温で２０時間攪拌した。この溶液について、１５０　ｍＭ食塩水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）、及び脱イオン水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（１２．０　ｍｇ，０．６９９　μｍｏｌ，収率ｑｕａｎｔ．）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．５２であった。ＩＣＰ－ＡＥＳ測定結果より、ＰＧｌｕの側鎖カルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、５６％であった。これは、１ポリマーに対して、約１３個の天然型Ｂ－アミノドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．４０７－２．６３（ｍ，３５６Ｈ），３．０３－３．２６（ｍ，３５Ｈ），３．５４－３．７８（ｍ，９０９Ｈ），４．０６－４．４５（ｍ，２２Ｈ）． The compound obtained in Synthesis Example 18 (10 mg, 0.734 μmol, Glu residue: 0.0169 mmol) was dissolved in DMSO (5.0 ml), and N-hydroxysuccinimide (9.7 mg, 0. 0845 mmol) and WSC · HCl (16.0 mg, 0.0845 mmol) were added, and the mixture was stirred at room temperature for 2 hours. Subsequently, the compound obtained in Synthesis Example 29 (29 mg, 0.847 mmol) and triethylamine (23.6 μl, 0.169 mmol) were added, followed by stirring at room temperature for 20 hours. This solution was subjected to three dialysis against 150 mM saline (total 20 hours, fractional molecular weight 6000 to 8000 Da) and three times against deionized water (total 20 hours, fractional molecular weight 6000 to 8000 Da). Purification and lyophilization gave the title compound (12.0 mg, 0.699 μmol, yield quant.) As a white powder. The molecular weight distribution (Mw / Mn) was 1.52 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction rate of natural B-dodecaborate with respect to the side chain carboxylic acid of PGlu was 56%. This corresponds to the introduction of about 13 natural B-aminododecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.407-2.63 (m, 356H), 3.03-3.26 (m, 35H), 3.54-3.78 (M, 909H), 4.06-4.45 (m, 22H).
合成例３０
ポリエチレングリコール（Ｍｗ＝２０，０００）－ベンジル保護ポリグルタミン酸（２２ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ（Ｂｎ）））の合成 Synthesis Example 30
Synthesis of polyethylene glycol (Mw = 20,000) -benzyl protected polyglutamic acid (22mer) -block copolymer (PEG-bP (Glu (Bn)))
Figure JPOXMLDOC01-appb-C000070
Figure JPOXMLDOC01-appb-C000070
　合成例１３と同様にして、Ｈ２Ｎ－ＰＥＧ－ＯＭｅ（Ｍｗ＝２０，０００，３．０　ｇ，０．１４２　ｍｍｏｌ）とＢＬＧ－ＮＣＡ（９３４　ｍｇ，３．５５　ｍｍｏｌ）より、標題化合物（３．２４　ｇ，０．１３３　ｍｍｏｌ，収率９４％）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤＣｌ３）により、得られた化合物の重合度が２２であることを確認した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤＣｌ３）δ（ｐｐｍ）：１．９７－２．７５（ｍ，８５Ｈ），３．５２－３．８２（ｍ，１８１８Ｈ），３．８９－４．０５（ｍ，２２Ｈ），４．９７－５．１９（ｍ，４４Ｈ），７．１０－７．４６（ｍ，１１０Ｈ）． In the same manner as in Synthesis Example 13, from the H 2 N-PEG-OMe (Mw = 20,000, 3.0 g, 0.142 mmol) and BLG-NCA (934 mg, 3.55 mmol), the title compound ( 3.24 g, 0.133 mmol, 94% yield) was obtained as a white powder. It was confirmed by 1 H-NMR (400 MHz, CDCl 3 ) that the polymerization degree of the obtained compound was 22.
1 H-NMR (400 MHz, CDCl 3 ) δ (ppm): 1.97-2.75 (m, 85H), 3.52-3.82 (m, 1818H), 3.89-4.05 ( m, 22H), 4.97-5.19 (m, 44H), 7.10-7.46 (m, 110H).
合成例３１
ポリエチレングリコール（Ｍｗ＝２０，０００）－ポリグルタミン酸（２２ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ））の合成 Synthesis Example 31
Synthesis of polyethylene glycol (Mw = 20,000) -polyglutamic acid (22mer) -block copolymer (PEG-bP (Glu))
Figure JPOXMLDOC01-appb-C000071
Figure JPOXMLDOC01-appb-C000071
　合成例３０で得られた化合物（３．２　ｇ，０．１３３　ｍｍｏｌ）を０．５Ｎ水酸化ナトリウム水溶液（２７　ｍｌ）中で１６時間攪拌した。この溶液を、脱イオン水に対する３回の透析（計６時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（２．１５　ｇ，０．０９２　ｍｍｏｌ，収率６５％）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）により、脱保護が完了したことを確認した。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．０６であった。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：１．７５－２．０９（ｍ，４３Ｈ），２．１１－２．３０（ｍ，４０Ｈ），３．５０－３．７５（ｍ，１８１８Ｈ），４．１５－４．３２（ｍ，１９Ｈ）． The compound obtained in Synthesis Example 30 (3.2 g, 0.133 mmol) was stirred in 0.5N aqueous sodium hydroxide solution (27 ml) for 16 hours. This solution was purified by dialysis 3 times against deionized water (6 hours in total, fractional molecular weight 6000 to 8000 Da), lyophilized to give the title compound (2.15 g, 0.092 mmol, yield 65 %) As a white powder. 1 H-NMR (400 MHz, D 2 O) was thus confirmed that the deprotection was complete. The molecular weight distribution (Mw / Mn) was 1.06 by size exclusion chromatography.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 1.75-2.09 (m, 43H), 2.11-2.30 (m, 40H), 3.50-3.75 (M, 1818H), 4.15-4.32 (m, 19H).
実施例１２
ポリエチレングリコール（Ｍｗ＝２０，０００）－（ドデカボレート－アミド結合）ポリグルタミン酸（２２ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｂ１２Ｈ１１））の合成 Example 12
Synthesis of polyethylene glycol (Mw = 20,000)-(dodecaborate-amide bond) polyglutamic acid (22mer) -block copolymer (PEG-bP (Glu-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000072
Figure JPOXMLDOC01-appb-C000072
　実施例５と同様にして、合成例３１で得られた化合物（２００　ｍｇ，０．５６７　μｍｏｌ，Ｇｌｕ残基：０．１８８　ｍｍｏｌ）と合成例２１で得られた化合物（３３１　ｍｇ，０．４７１　ｍｍｏｌ）をＤＭＳＯ中で縮合させた。この溶液について、１５０　ｍＭ食塩水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）、脱イオン水に対する３回の透析（計２０時間、分画分子量６０００～８０００　Ｄａ）、及び１００　ｍＭリン酸緩衝液（ｐＨ８．０）に対する２回の透析（計６時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（２１９　ｍｇ，８．３１　μｍｏｌ，収率９７％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．１２であった。ＩＣＰ－ＡＥＳ測定結果より、ポリグルタミン酸（ＰＧｌｕ）の側鎖のカルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、５５％であった。これは、１ポリマーに対して、約１２個の天然型Ｂ－ドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．２０－２．４８（ｍ，１７０Ｈ），２．１９－２．４８（ｍ，３１Ｈ），３．１８－３．４０（ｍ，３３Ｈ），３．５０－３．８０（ｍ，１９１０Ｈ），４．１５－４．４５（ｍ，１７Ｈ）． In the same manner as in Example 5, the compound obtained in Synthesis Example 31 (200 mg, 0.567 μmol, Glu residue: 0.188 mmol) and the compound obtained in Synthesis Example 21 (331 mg, 0.471). mmol) was condensed in DMSO. For this solution, three dialysis against 150 mM saline (total 20 hours, fractional molecular weight 6000-8000 Da), three dialysis against deionized water (total 20 hours, fractional molecular weight 6000-8000 Da), and The product was purified by dialysis twice against a 100 mM phosphate buffer (pH 8.0) (total 6 hours, molecular weight cut off: 6000 to 8000 Da), lyophilized, and the title compound (219 mg, 8.31 μmol) was collected. 97%) was obtained as a white powder. The molecular weight distribution (Mw / Mn) was 1.12 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction rate of natural B-dodecaborate with respect to the carboxylic acid in the side chain of polyglutamic acid (PGlu) was 55%. This corresponds to the introduction of about 12 natural B-dodecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.20-2.48 (m, 170H), 2.19-2.48 (m, 31H), 3.18-3.40 (M, 33H), 3.50-3.80 (m, 1910H), 4.15-4.45 (m, 17H).
合成例３２
ポリエチレングリコール（Ｍｗ＝２０，０００）－ベンジル保護ポリグルタミン酸（４３ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ（Ｂｎ）））の合成 Synthesis Example 32
Synthesis of polyethylene glycol (Mw = 20,000) -benzyl protected polyglutamic acid (43mer) -block copolymer (PEG-bP (Glu (Bn)))
Figure JPOXMLDOC01-appb-C000073
Figure JPOXMLDOC01-appb-C000073
　合成例１３と同様にして、Ｈ２Ｎ－ＰＥＧ－ＯＭｅ（Ｍｗ＝２０，０００，３．２　ｇ，０．１５５　ｍｍｏｌ）とＢＬＧ－ＮＣＡ（２．０　ｇ，５．３５　ｍｍｏｌ）より、標題化合物（４．６　ｇ，０．１５６　ｍｍｏｌ，収率ｑｕａｎｔ．）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤＣｌ３）により、得られた化合物の重合度が４３であることを確認した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤＣｌ３）δ（ｐｐｍ）：１．９８－２．８１（ｍ，１６５Ｈ），３．５５－３．８０（ｍ，１８１８Ｈ），３．８９－４．１９（ｍ，２６Ｈ），４．９０－５．１１（ｍ，８９Ｈ），７．１５－７．４６（ｍ，２２０Ｈ）． In the same manner as in Synthesis Example 13, from H 2 N-PEG-OMe (Mw = 20,000, 3.2 g, 0.155 mmol) and BLG-NCA (2.0 g, 5.35 mmol) The compound (4.6 g, 0.156 mmol, yield quant.) Was obtained as a white powder. It was confirmed by 1 H-NMR (400 MHz, CDCl 3 ) that the degree of polymerization of the obtained compound was 43.
1 H-NMR (400 MHz, CDCl 3 ) δ (ppm): 1.98-2.81 (m, 165H), 3.55-3.80 (m, 1818H), 3.89-4.19 ( m, 26H), 4.90-5.11 (m, 89H), 7.15-7.46 (m, 220H).
合成例３３
ポリエチレングリコール（Ｍｗ＝２０，０００）－ポリグルタミン酸（４３ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ））の合成 Synthesis Example 33
Synthesis of polyethylene glycol (Mw = 20,000) -polyglutamic acid (43mer) -block copolymer (PEG-bP (Glu))
Figure JPOXMLDOC01-appb-C000074
Figure JPOXMLDOC01-appb-C000074
　合成例３２で得られた化合物（４．６　ｇ，０．１５５　ｍｍｏｌ）を０．５Ｎ水酸化ナトリウム水溶液（６７　ｍｌ）中で１６時間攪拌した。この溶液を、脱イオン水に対する３回の透析（計６時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（２．７７　ｇ，０．０１０４　ｍｍｏｌ，収率６７％）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）により、脱保護が完了したことを確認した。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．０６であった。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：１．７９－２．１０（ｍ，８０Ｈ），２．１０－２．３１（ｍ，７７Ｈ），３．５２－３．７９（ｍ，１８１８Ｈ），４．１５－４．３５（ｍ，４１Ｈ）． The compound obtained in Synthesis Example 32 (4.6 g, 0.155 mmol) was stirred in 0.5N aqueous sodium hydroxide solution (67 ml) for 16 hours. This solution was purified by dialysis 3 times against deionized water (total 6 hours, molecular weight cut off 6000 to 8000 Da), freeze-dried, and the title compound (2.77 g, 0.0104 mmol, yield 67). %) As a white powder. 1 H-NMR (400 MHz, D 2 O) confirmed the completion of deprotection. The molecular weight distribution (Mw / Mn) was 1.06 by size exclusion chromatography.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 1.79-2.10 (m, 80H), 2.10-2.31 (m, 77H), 3.52-3.79 (M, 1818H), 4.15-4.35 (m, 41H).
実施例１３
ポリエチレングリコール（Ｍｗ＝２０，０００）－（ドデカボレート－アミド結合）ポリグルタミン酸（４３ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｂ１２Ｈ１１））の合成 Example 13
Synthesis of polyethylene glycol (Mw = 20,000)-(dodecaborate-amide bond) polyglutamic acid (43mer) -block copolymer (PEG-bP (Glu-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000075
Figure JPOXMLDOC01-appb-C000075
　実施例１２と同様にして、合成例３３で得られた化合物（２００　ｍｇ，７．５４　μｍｏｌ，Ｇｌｕ残基：０．３２４　ｍｍｏｌ）と合成例２１で得られた化合物（５６９　ｍｇ，０．８１０　ｍｍｏｌ）より、標題化合物（２４８　ｍｇ，７．２４　μｍｏｌ，収率９６％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．１７であった。ＩＣＰ－ＡＥＳ測定結果より、ポリグルタミン酸（ＰＧｌｕ）の側鎖のカルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、７２％であった。これは、１ポリマーに対して、約３１個の天然型Ｂ－ドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．３５－２．１８（ｍ，３７３Ｈ），２．２０－２．４７（ｍ，６３Ｈ），３．２０－３．４２（ｍ，７３Ｈ），３．５０－３．６５（ｍ，２０２９Ｈ），４．１５－４．４０（ｍ，２８Ｈ）． In the same manner as in Example 12, the compound obtained in Synthesis Example 33 (200 mg, 7.54 μmol, Glu residue: 0.324 mmol) and the compound obtained in Synthesis Example 21 (569 mg, 0.810 mmol) gave the title compound (248 mg, 7.24 μmol, yield 96%) as a white powder. The molecular weight distribution (Mw / Mn) was 1.17 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction rate of natural B-dodecaborate with respect to the carboxylic acid in the side chain of polyglutamic acid (PGlu) was 72%. This corresponds to the introduction of about 31 natural B-dodecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.35-2.18 (m, 373H), 2.20-2.47 (m, 63H), 3.20-3.42 (M, 73H), 3.50-3.65 (m, 2029H), 4.15-4.40 (m, 28H).
合成例３４
ポリエチレングリコール（Ｍｗ＝２０，０００）－ベンジル保護ポリグルタミン酸（９７ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ（Ｂｎ）））の合成 Synthesis Example 34
Synthesis of polyethylene glycol (Mw = 20,000) -benzyl protected polyglutamic acid (97mer) -block copolymer (PEG-bP (Glu (Bn)))
Figure JPOXMLDOC01-appb-C000076
Figure JPOXMLDOC01-appb-C000076
　合成例１３と同様にして、Ｈ２Ｎ－ＰＥＧ－ＯＭｅ（Ｍｗ＝２０，０００，５００　ｍｇ，０．０２５　ｍｍｏｌ）とＢＬＧ－ＮＣＡ（７２４　ｍｇ，２．７５　ｍｍｏｌ）をＤＭＦ（１Ｍチオウレア含有）中にて重合させ、標題化合物を白色粉体として得た。 In the same manner as in Synthesis Example 13, H 2 N-PEG-OMe (Mw = 20,000, 500 mg, 0.025 mmol) and BLG-NCA (724 mg, 2.75 mmol) were added to DMF (containing 1M thiourea). Polymerized in to give the title compound as a white powder.
合成例３５
ポリエチレングリコール（Ｍｗ＝２０，０００）－ポリグルタミン酸（９７ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ））の合成 Synthesis Example 35
Synthesis of polyethylene glycol (Mw = 20,000) -polyglutamic acid (97mer) -block copolymer (PEG-bP (Glu))
Figure JPOXMLDOC01-appb-C000077
Figure JPOXMLDOC01-appb-C000077
　合成例３４で得られた化合物を０．５Ｎ水酸化ナトリウム水溶液（１９　ｍｌ）中で１６時間攪拌した。この溶液を、脱イオン水に対する３回の透析（計６時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（３９５　ｍｇ，０．０１１２　ｍｍｏｌ，２段階収率４５％）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）により、脱保護の完了、及び重合度が９７であることを確認した。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．１２であった。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：１．７９－２．０９（ｍ，２０４Ｈ），２．１０－２．３２（ｍ，１９５Ｈ），３．５１－３．７９（ｍ，１８１８Ｈ），４．１８－４．３９（ｍ，９４Ｈ）． The compound obtained in Synthesis Example 34 was stirred in 0.5N aqueous sodium hydroxide solution (19 ml) for 16 hours. This solution was purified by dialysis 3 times against deionized water (total 6 hours, molecular weight cut off 6000-8000 Da), lyophilized, and the title compound (395 mg, 0.0112 mmol, 2-step yield 45 %) As a white powder. 1 H-NMR (400 MHz, D 2 O) confirmed the completion of deprotection and the degree of polymerization was 97. The molecular weight distribution (Mw / Mn) was 1.12 by size exclusion chromatography.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 1.79-2.09 (m, 204H), 2.10-2.32 (m, 195H), 3.51-3.79 (M, 1818H), 4.18-4.39 (m, 94H).
実施例１４
ポリエチレングリコール（Ｍｗ＝２０，０００）－（ドデカボレート－アミド結合）ポリグルタミン酸（９７ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｂ１２Ｈ１１））の合成 Example 14
Synthesis of polyethylene glycol (Mw = 20,000)-(dodecaborate-amide bond) polyglutamic acid (97mer) -block copolymer (PEG-bP (Glu-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000078
Figure JPOXMLDOC01-appb-C000078
　実施例１２と同様にして、合成例３５で得られた化合物（１２０　ｍｇ，３．５２　μｍｏｌ，Ｇｌｕ残基：０．３４２　ｍｍｏｌ）と合成例２１で得られた化合物（６２４　ｍｇ，０．８５５　ｍｍｏｌ）より、標題化合物（１４１　ｍｇ，７．２４　μｍｏｌ，収率８１％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．１０であった。ＩＣＰ－ＡＥＳ測定結果より、ポリグルタミン酸（ＰＧｌｕ）の側鎖のカルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、６１％であった。これは、１ポリマーに対して、約５９個の天然型Ｂ－ドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．２２－２．２３（ｍ，９９９Ｈ），２．２３－２．４８（ｍ，１８５Ｈ），３．１８－３．４３（ｍ，２２７Ｈ），３．６４－３．８７（ｍ，２３３２Ｈ），４．１６－４．４６（ｍ，８１Ｈ）． In the same manner as in Example 12, the compound obtained in Synthesis Example 35 (120 mg, 3.52 μmol, Glu residue: 0.342 mmol) and the compound obtained in Synthesis Example 21 (624 mg, 0.855 mmol) gave the title compound (141 mg, 7.24 μmol, 81% yield) as a white powder. The molecular weight distribution (Mw / Mn) was 1.10 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction rate of natural B-dodecaborate with respect to the carboxylic acid in the side chain of polyglutamic acid (PGlu) was 61%. This corresponds to the introduction of about 59 natural B-dodecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.22-2.23 (m, 999H), 2.23-2.48 (m, 185H), 3.18-3.43 (M, 227H), 3.64-3.87 (m, 2332H), 4.16-4.46 (m, 81H).
合成例３６
ポリエチレングリコール（Ｍｗ＝３０，０００）－ベンジル保護ポリグルタミン酸（２２ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ（Ｂｎ）））の合成 Synthesis Example 36
Synthesis of polyethylene glycol (Mw = 30,000) -benzyl protected polyglutamic acid (22mer) -block copolymer (PEG-bP (Glu (Bn)))
Figure JPOXMLDOC01-appb-C000079
Figure JPOXMLDOC01-appb-C000079
　合成例１３と同様にして、Ｈ２Ｎ－ＰＥＧ－ＯＭｅ（Ｍｗ＝３０，０００，５００　ｍｇ，０．０１６７　ｍｍｏｌ）とＢＬＧ－ＮＣＡ（１１０　ｍｇ，０．４１７　ｍｍｏｌ）より、標題化合物（５４７　ｍｇ，０．０１５７ｍｍｏｌ，収率９４％）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤＣｌ３）により、得られた化合物の重合度が２２であることを確認した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤＣｌ３）δ（ｐｐｍ）：１．９８－２．７０（ｍ，８０Ｈ），３．５５－３．８０（ｍ，２７２７Ｈ），３．８９－４．０５（ｍ，１６Ｈ），４．９２－５．２２（ｍ，４３Ｈ），７．１２－７．４０（ｍ，１０８Ｈ）． In the same manner as in Synthesis Example 13, from the H 2 N-PEG-OMe (Mw = 30,000,500 mg, 0.0167 mmol) and BLG-NCA (110 mg, 0.417 mmol), the title compound (547 mg , 0.0157 mmol, 94% yield) as a white powder. It was confirmed by 1 H-NMR (400 MHz, CDCl 3 ) that the polymerization degree of the obtained compound was 22.
1 H-NMR (400 MHz, CDCl 3 ) δ (ppm): 1.98-2.70 (m, 80H), 3.55-3.80 (m, 2727H), 3.89-4.05 ( m, 16H), 4.92-5.22 (m, 43H), 7.12-7.40 (m, 108H).
合成例３７
ポリエチレングリコール（Ｍｗ＝３０，０００）－ポリグルタミン酸（２２ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ））の合成 Synthesis Example 37
Synthesis of polyethylene glycol (Mw = 30,000) -polyglutamic acid (22mer) -block copolymer (PEG-bP (Glu))
Figure JPOXMLDOC01-appb-C000080
Figure JPOXMLDOC01-appb-C000080
　合成例３６で得られた化合物（５４７　ｍｇ,　０．０１５７　ｍｍｏｌ）を０．５Ｎ水酸化ナトリウム水溶液（４　ｍｌ）中で１６時間攪拌した。この溶液を、脱イオン水に対する３回の透析（計６時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（５０８　ｍｇ，０．０１５３　ｍｍｏｌ，収率９７％）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）により、脱保護が完了したことを確認した。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．０８であった。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：１．８０－２．０９（ｍ，４０Ｈ），２．０１－２．３２（ｍ，３９Ｈ），３．４９－３．７４（ｍ，２７２７Ｈ），４．１９－４．３９（ｍ，１５Ｈ）． The compound obtained in Synthesis Example 36 (547 mg, 0.0157 mmol) was stirred in 0.5N aqueous sodium hydroxide solution (4 ml) for 16 hours. This solution was purified by dialysis three times against deionized water (total 6 hours, molecular weight cut off 6000 to 8000 Da), freeze-dried, and the title compound (508 mg, 0.0153 mmol, yield 97%) Was obtained as a white powder. 1 H-NMR (400 MHz, D 2 O) was thus confirmed that the deprotection was complete. The molecular weight distribution (Mw / Mn) was 1.08 by size exclusion chromatography.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 1.80-2.09 (m, 40H), 2.01-2.32 (m, 39H), 3.49-3.74 (M, 2727H), 4.19-4.39 (m, 15H).
実施例１５
ポリエチレングリコール（Ｍｗ＝３０，０００）－（ドデカボレート－アミド結合）ポリグルタミン酸（２２ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｂ１２Ｈ１１））の合成 Example 15
Synthesis of polyethylene glycol (Mw = 30,000)-(dodecaborate-amide bond) polyglutamic acid (22mer) -block copolymer (PEG-bP (Glu-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000081
Figure JPOXMLDOC01-appb-C000081
　実施例１２と同様にして、合成例３７で得られた化合物（２５０　ｍｇ，７．５０　μｍｏｌ，Ｇｌｕ残基：０．１６５　ｍｍｏｌ）と合成例２１で得られた化合物（３０１　ｍｇ，０．４１２　ｍｍｏｌ）より、標題化合物（２３３　ｍｇ，６．４１　μｍｏｌ，収率８５％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．１６であった。ＩＣＰ－ＡＥＳ測定結果より、ポリグルタミン酸（ＰＧｌｕ）の側鎖のカルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、５５％であった。これは、１ポリマーに対して、約１２個の天然型Ｂ－ドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．３１－２．１８（ｍ，１５５Ｈ），２．１８－２．４２（ｍ，２８Ｈ），３．２０－３．３９（ｍ，３３Ｈ），３．４８－３．７７（ｍ，２８２２Ｈ），４．１７－４．４２（ｍ，１１Ｈ）． In the same manner as in Example 12, the compound obtained in Synthesis Example 37 (250 mg, 7.50 μmol, Glu residue: 0.165 mmol) and the compound obtained in Synthesis Example 21 (301 mg, 0.412) mmol) gave the title compound (233 mg, 6.41 μmol, yield 85%) as a white powder. The molecular weight distribution (Mw / Mn) was 1.16 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction rate of natural B-dodecaborate with respect to the carboxylic acid in the side chain of polyglutamic acid (PGlu) was 55%. This corresponds to the introduction of about 12 natural B-dodecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.31-2.18 (m, 155H), 2.18-2.42 (m, 28H), 3.20-3.39 (M, 33H), 3.48-3.77 (m, 2822H), 4.17-4.42 (m, 11H).
合成例３８
ポリエチレングリコール（Ｍｗ＝３０，０００）－ベンジル保護ポリグルタミン酸（３９ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ（Ｂｎ）））の合成 Synthesis Example 38
Synthesis of polyethylene glycol (Mw = 30,000) -benzyl protected polyglutamic acid (39mer) -block copolymer (PEG-bP (Glu (Bn)))
Figure JPOXMLDOC01-appb-C000082
Figure JPOXMLDOC01-appb-C000082
　合成例１３と同様にして、Ｈ２Ｎ－ＰＥＧ－ＯＭｅ（Ｍｗ＝３０，０００，５００　ｍｇ，０．０１６７　ｍｍｏｌ）とＢＬＧ－ＮＣＡ（２２０　ｍｇ，０．８３５　ｍｍｏｌ）より、標題化合物（６１０　ｍｇ，０．０１５８ｍｍｏｌ，収率９５％）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤＣｌ３）により、得られた化合物の重合度が３９であることを確認した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤＣｌ３）δ（ｐｐｍ）：１．９８－２．８９（ｍ，１４８Ｈ），３．５１－３．８０（ｍ，２７２７Ｈ），３．８０－４．２０（ｍ，３５Ｈ），４．９０－５．２５（ｍ，７７Ｈ），７．１２－７．４０（ｍ，１９３Ｈ）． In the same manner as in Synthesis Example 13, from the H 2 N-PEG-OMe (Mw = 30,000, 500 mg, 0.0167 mmol) and BLG-NCA (220 mg, 0.835 mmol), the title compound (610 mg , 0.0158 mmol, 95% yield) as a white powder. It was confirmed by 1 H-NMR (400 MHz, CDCl 3 ) that the obtained compound had a degree of polymerization of 39.
1 H-NMR (400 MHz, CDCl 3 ) δ (ppm): 1.98-2.89 (m, 148H), 3.51-3.80 (m, 2727H), 3.80-4.20 ( m, 35H), 4.90-5.25 (m, 77H), 7.12-7.40 (m, 193H).
合成例３９
ポリエチレングリコール（Ｍｗ＝３０，０００）－ポリグルタミン酸（３９ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ））の合成 Synthesis Example 39
Synthesis of polyethylene glycol (Mw = 30,000) -polyglutamic acid (39mer) -block copolymer (PEG-bP (Glu))
Figure JPOXMLDOC01-appb-C000083
Figure JPOXMLDOC01-appb-C000083
　合成例３８で得られた化合物を０．５Ｎ水酸化ナトリウム水溶液（６　ｍｌ）中で１６時間攪拌した。この溶液を、脱イオン水に対する３回の透析（計６時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（４５２　ｍｇ，０．０１２６　ｍｍｏｌ，収率７５％）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）により、脱保護が完了したことを確認した。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．０５であった。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：１．７５－２．０９（ｍ，７１Ｈ），２．０９－２．３１（ｍ，７０Ｈ），３．５０－３．７８（ｍ，２７２７Ｈ），４．１０－４．３８（ｍ，３５Ｈ）． The compound obtained in Synthesis Example 38 was stirred in 0.5N aqueous sodium hydroxide solution (6 ml) for 16 hours. This solution was purified by dialysis three times against deionized water (total 6 hours, molecular weight cut off 6000 to 8000 Da), lyophilized, and titled compound (452 mg, 0.0126 mmol, yield 75%) Was obtained as a white powder. 1 H-NMR (400 MHz, D 2 O) was thus confirmed that the deprotection was complete. The molecular weight distribution (Mw / Mn) was 1.05 by size exclusion chromatography.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 1.75-2.09 (m, 71H), 2.09-2.31 (m, 70H), 3.50-3.78 (M, 2727H), 4.10-4.38 (m, 35H).
実施例１６
ポリエチレングリコール（Ｍｗ＝３０，０００）－（ドデカボレート－アミド結合）ポリグルタミン酸（３９ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｂ１２Ｈ１１））の合成 Example 16
Synthesis of polyethylene glycol (Mw = 30,000)-(dodecaborate-amide bond) polyglutamic acid (39mer) -block copolymer (PEG-bP (Glu-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000084
Figure JPOXMLDOC01-appb-C000084
　実施例１２と同様にして、合成例３９で得られた化合物（１５０　ｍｇ，４．１８　μｍｏｌ，Ｇｌｕ残基：０．１６３　ｍｍｏｌ）と合成例２１で得られた化合物（２９７　ｍｇ，０．４０７　ｍｍｏｌ）より、標題化合物（１４０　ｍｇ，３．３６　μｍｏｌ，収率８０％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．２４であった。ＩＣＰ－ＡＥＳ測定結果より、ポリグルタミン酸（ＰＧｌｕ）の側鎖のカルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、５９％であった。これは、１ポリマーに対して、約２３個の天然型Ｂ－ドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．１０－２．１５（ｍ，４２２Ｈ），２．１５－２．４７（ｍ，７１Ｈ），３．２０－３．４０（ｍ，６６Ｈ），３．４９－３．７８（ｍ，２９１５Ｈ），４．１５－４．４５（ｍ，３７Ｈ）． In the same manner as in Example 12, the compound obtained in Synthesis Example 39 (150 mg, 4.18 μmol, Glu residue: 0.163 mmol) and the compound obtained in Synthesis Example 21 (297 mg, 0.407) mmol) gave the title compound (140 mg, 3.36 μmol, 80% yield) as a white powder. The molecular weight distribution (Mw / Mn) was 1.24 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction rate of natural B-dodecaborate with respect to the carboxylic acid in the side chain of polyglutamic acid (PGlu) was 59%. This corresponds to the introduction of about 23 natural B-dodecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.10-2.15 (m, 422H), 2.15-2.47 (m, 71H), 3.20-3.40 (M, 66H), 3.49-3.78 (m, 2915H), 4.15-4.45 (m, 37H).
合成例４０
ポリエチレングリコール（Ｍｗ＝４０，０００）－ベンジル保護ポリグルタミン酸（２５ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ（Ｂｎ）））の合成 Synthesis Example 40
Synthesis of polyethylene glycol (Mw = 40,000) -benzyl protected polyglutamic acid (25mer) -block copolymer (PEG-bP (Glu (Bn)))
Figure JPOXMLDOC01-appb-C000085
Figure JPOXMLDOC01-appb-C000085
　合成例１３と同様にして、Ｈ２Ｎ－ＰＥＧ－ＯＭｅ（Ｍｗ＝３０，０００，１．０　ｇ，０．０２５　ｍｍｏｌ）とＢＬＧ－ＮＣＡ（１６５　ｍｇ，０．６２５　ｍｍｏｌ）より、標題化合物（１．２　ｇ,０．０２６４ｍｍｏｌ，収率ｑｕａｎｔ．）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤＣｌ３）により、得られた化合物の重合度が２５であることを確認した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤＣｌ３）δ（ｐｐｍ）：１．９８－２．８９（ｍ，１００Ｈ），３．５１－３．８０（ｍ，３６３６Ｈ），３．８９－４．１１（ｍ，１９Ｈ），４．８９－５．１８（ｍ，５１Ｈ），７．１５－７．４０（ｍ，１３０Ｈ）． In the same manner as in Synthesis Example 13, from the H 2 N-PEG-OMe (Mw = 30,000, 1.0 g, 0.025 mmol) and BLG-NCA (165 mg, 0.625 mmol), the title compound ( 1.2 g, 0.0264 mmol, yield quant.) Was obtained as a white powder. It was confirmed by 1 H-NMR (400 MHz, CDCl 3 ) that the degree of polymerization of the obtained compound was 25.
1 H-NMR (400 MHz, CDCl 3 ) δ (ppm): 1.98-2.89 (m, 100H), 3.51-3.80 (m, 3636H), 3.89-4.11 ( m, 19H), 4.89-5.18 (m, 51H), 7.15-7.40 (m, 130H).
合成例４１
ポリエチレングリコール（Ｍｗ＝４０，０００）－ポリグルタミン酸（２５ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ））の合成 Synthesis Example 41
Synthesis of polyethylene glycol (Mw = 40,000) -polyglutamic acid (25mer) -block copolymer (PEG-bP (Glu))
Figure JPOXMLDOC01-appb-C000086
Figure JPOXMLDOC01-appb-C000086
　合成例４０で得られた化合物を０．５Ｎ水酸化ナトリウム水溶液（７　ｍｌ）中で１６時間攪拌した。この溶液を、脱イオン水に対する３回の透析（計６時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（９５８　ｍｇ，０．０２１８　ｍｍｏｌ，収率８６％）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）により、脱保護が完了したことを確認した。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．１１であった。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：１．７４－２．０９（ｍ，４９Ｈ），２．０９－２．２９（ｍ，４８Ｈ），３．５２－３．７５（ｍ，３６３６Ｈ），４．１５－４．３３（ｍ，２５Ｈ）． The compound obtained in Synthesis Example 40 was stirred in 0.5N aqueous sodium hydroxide solution (7 ml) for 16 hours. This solution was purified by dialysis three times against deionized water (total 6 hours, molecular weight cut off 6000 to 8000 Da), lyophilized, and titled compound (958 mg, 0.0218 mmol, yield 86%). Was obtained as a white powder. 1 H-NMR (400 MHz, D 2 O) was thus confirmed that the deprotection was complete. The molecular weight distribution (Mw / Mn) was 1.11 by size exclusion chromatography.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 1.74-2.09 (m, 49H), 2.09-2.29 (m, 48H), 3.52-3.75 (M, 3636H), 4.15-4.33 (m, 25H).
実施例１７
ポリエチレングリコール（Ｍｗ＝４０，０００）－（ドデカボレート－アミド結合）ポリグルタミン酸（２５ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｂ１２Ｈ１１））の合成 Example 17
Synthesis of polyethylene glycol (Mw = 40,000)-(dodecaborate-amide bond) polyglutamic acid (25mer) -block copolymer (PEG-bP (Glu-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000087
Figure JPOXMLDOC01-appb-C000087
　実施例１２と同様にして、合成例４１で得られた化合物（３００　ｍｇ，７．０８　μｍｏｌ，Ｇｌｕ残基：０．１７７　ｍｍｏｌ）と合成例２１で得られた化合物（３２４　ｍｇ，０．４４４　ｍｍｏｌ）より、標題化合物（２７６　ｍｇ，５．７７　μｍｏｌ，収率８１％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．０９であった。ＩＣＰ－ＡＥＳ測定結果より、ポリグルタミン酸（ＰＧｌｕ）の側鎖のカルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、６２％であった。これは、１ポリマーに対して、約１６個の天然型Ｂ－ドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．１０－２．２８（ｍ，１９３Ｈ），２．２８－２．５２（ｍ，３５Ｈ），３．２３－３．４３（ｍ，３７Ｈ），３．５０－３．８１（ｍ，３７４８Ｈ），４．２０－４．４９（ｍ，１６Ｈ）． In the same manner as in Example 12, the compound obtained in Synthesis Example 41 (300 mg, 7.08 μmol, Glu residue: 0.177 mmol) and the compound obtained in Synthesis Example 21 (324 mg, 0.444) mmol) gave the title compound (276 mg, 5.77 μmol, 81% yield) as a white powder. The molecular weight distribution (Mw / Mn) was 1.09 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction rate of natural B-dodecaborate with respect to the carboxylic acid in the side chain of polyglutamic acid (PGlu) was 62%. This corresponds to the introduction of about 16 natural B-dodecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.10-2.28 (m, 193H), 2.28-2.52 (m, 35H), 3.23-3-43 (M, 37H), 3.50-3.81 (m, 3748H), 4.20-4.49 (m, 16H).
合成例４２
ポリエチレングリコール（Ｍｗ＝４０，０００）－ベンジル保護ポリグルタミン酸（３７ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ（Ｂｎ）））の合成 Synthesis Example 42
Synthesis of polyethylene glycol (Mw = 40,000) -benzyl protected polyglutamic acid (37mer) -block copolymer (PEG-bP (Glu (Bn)))
Figure JPOXMLDOC01-appb-C000088
Figure JPOXMLDOC01-appb-C000088
　合成例１３と同様にして、Ｈ２Ｎ－ＰＥＧ－ＯＭｅ（Ｍｗ＝４０，０００，５００　ｍｇ，０．０１２５　ｍｍｏｌ）とＢＬＧ－ＮＣＡ（１６５　ｍｇ，０．６２５　ｍｍｏｌ）より、標題化合物（５５２　ｍｇ，０．０１１５ｍｍｏｌ，収率９２％）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤＣｌ３）により、得られた化合物の重合度が３７であることを確認した。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，ＣＤＣｌ３）δ（ｐｐｍ）：１．９８－２．８９（ｍ，１４２Ｈ），３．５１－３．８０（ｍ，３６３６Ｈ），３．８９－４．１１（ｍ，３８Ｈ），４．８９－５．１８（ｍ，７５Ｈ），７．１５－７．４０（ｍ，１８５Ｈ）． In the same manner as in Synthesis Example 13, the title compound (552 mg) was prepared from H 2 N-PEG-OMe (Mw = 40,000,500 mg, 0.0125 mmol) and BLG-NCA (165 mg, 0.625 mmol). , 0.0115 mmol, 92% yield) as a white powder. It was confirmed by 1 H-NMR (400 MHz, CDCl 3 ) that the obtained compound had a degree of polymerization of 37.
1 H-NMR (400 MHz, CDCl 3 ) δ (ppm): 1.98-2.89 (m, 142H), 3.51-3.80 (m, 3636H), 3.89-4.11 ( m, 38H), 4.89-5.18 (m, 75H), 7.15-7.40 (m, 185H).
合成例４３
ポリエチレングリコール（Ｍｗ＝４０，０００）－ポリグルタミン酸（３７ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ））の合成 Synthesis Example 43
Synthesis of polyethylene glycol (Mw = 40,000) -polyglutamic acid (37mer) -block copolymer (PEG-bP (Glu))
Figure JPOXMLDOC01-appb-C000089
Figure JPOXMLDOC01-appb-C000089
　合成例４２で得られた化合物を０．５Ｎ水酸化ナトリウム水溶液（７　ｍｌ）中で１６時間攪拌した。この溶液を、脱イオン水に対する３回の透析（計６時間、分画分子量６０００～８０００　Ｄａ）により精製し、凍結乾燥を行い、標題化合物（５１１　ｍｇ，０．０１１２　ｍｍｏｌ，収率９７％）を白色粉体として得た。１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）により、脱保護が完了したことを確認した。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．０６であった。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：１．７５－２．０９（ｍ，７１Ｈ），２．０９－２．３２（ｍ，７１Ｈ），３．５３－３．７７（ｍ，３６３６Ｈ），４．１６－４．３６（ｍ，３６Ｈ）． The compound obtained in Synthesis Example 42 was stirred in 0.5N aqueous sodium hydroxide solution (7 ml) for 16 hours. This solution was purified by dialysis 3 times against deionized water (total 6 hours, molecular weight cut off 6000 to 8000 Da), freeze-dried, and the title compound (511 mg, 0.0112 mmol, yield 97%) Was obtained as a white powder. 1 H-NMR (400 MHz, D 2 O) was thus confirmed that the deprotection was complete. The molecular weight distribution (Mw / Mn) was 1.06 by size exclusion chromatography.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 1.75-2.09 (m, 71H), 2.09-2.32 (m, 71H), 3.53-3.77 (M, 3636H), 4.16-4.36 (m, 36H).
実施例１８
ポリエチレングリコール（Ｍｗ＝４０，０００）－（ドデカボレート－アミド結合）ポリグルタミン酸（３７ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－Ｂ１２Ｈ１１））の合成 Example 18
Synthesis of polyethylene glycol (Mw = 40,000)-(dodecaborate-amide bond) polyglutamic acid (37mer) -block copolymer (PEG-bP (Glu-B 12 H 11 ))
Figure JPOXMLDOC01-appb-C000090
Figure JPOXMLDOC01-appb-C000090
　実施例１２と同様にして、合成例４３で得られた化合物（１８０　ｍｇ，３．９４　μｍｏｌ，Ｇｌｕ残基：０．１４６　ｍｍｏｌ）と合成例２１で得られた化合物（２６６　ｍｇ，０．３６５　ｍｍｏｌ）より、標題化合物（１８６　ｍｇ，３．６４　μｍｏｌ，収率９２％）を白色粉体として得た。サイズ排除クロマトグラフィーにより、分子量分布（Ｍｗ／Ｍｎ）は１．０９であった。ＩＣＰ－ＡＥＳ測定結果より、ポリグルタミン酸（ＰＧｌｕ）の側鎖のカルボン酸に対する天然型Ｂ－ドデカボレートの導入率は、５９％であった。これは、１ポリマーに対して、約２２個の天然型Ｂ－ドデカボレートが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．１６－２．１７（ｍ，２５７Ｈ），２．１７－２．４２（ｍ，４５Ｈ），３．２３－３．４０（ｍ，４５Ｈ），３．４８－３．７８（ｍ,　３７７６Ｈ），４．１７－４．４２（ｍ，２６Ｈ）． In the same manner as in Example 12, the compound obtained in Synthesis Example 43 (180 mg, 3.94 μmol, Glu residue: 0.146 mmol) and the compound obtained in Synthesis Example 21 (266 mg, 0.365) mmol) gave the title compound (186 mg, 3.64 μmol, 92% yield) as a white powder. The molecular weight distribution (Mw / Mn) was 1.09 by size exclusion chromatography. From the results of ICP-AES measurement, the introduction rate of natural B-dodecaborate with respect to the carboxylic acid in the side chain of polyglutamic acid (PGlu) was 59%. This corresponds to the introduction of about 22 natural B-dodecaborates per polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.16-2.17 (m, 257H), 2.17-2.42 (m, 45H), 3.23-3.40 (M, 45H), 3.48-3.78 (m, 3776H), 4.17-4.42 (m, 26H).
実施例１９
ポリエチレングリコール（Ｍｎ＝１０，０００）－ＢＳＨ結合ポリグルタミン酸（４１ｍｅｒ）－ブロック共重合体（ＰＥＧ－ｂ－Ｐ（Ｇｌｕ－ＢＳＨ））の合成 Example 19
Synthesis of polyethylene glycol (Mn = 10,000) -BSH-linked polyglutamic acid (41mer) -block copolymer (PEG-bP (Glu-BSH))
Figure JPOXMLDOC01-appb-C000091
Figure JPOXMLDOC01-appb-C000091
　合成例２と同様の方法で合成した、ポリエチレングリコール（Ｍｎ＝１０，０００）－チオレート化ポリグルタミン酸（４１ｍｅｒ）－ブロック共重合体（２５０　ｍｇ，０．０１４　ｍｍｏｌ）と１０Ｂ－ＢＳＨ（１５０　ｍｇ，ブロック共重合体中のチオレート化された官能基に対して２　ｅｑ）を、１０　ｍＭ　ＨＥＰＥＳ（４－（２－ヒドロキシエチル）－１－ピペラジンエタンスルホン酸）（ｐＨ　８．０）／５００　ｍＭ食塩水（１５　ｍｌ）に溶解し、室温で３日間撹拌した。反応液を１５０　ｍＭ食塩水中で３回透析後、脱イオン水中で３回透析し（分画分子量３５００　Ｄａ）、凍結乾燥することで、標題化合物（２６６　ｍｇ，収率９６％）を得た。１０Ｂ－ＢＳＨの導入率は、１Ｈ－ＮＭＲ分析（Ｄ２Ｏ）により、ホウ素クラスターのプロトン（－ＢＨ；δ（ｐｐｍ）：０．５-２．０）とポリアミノ酸の重合度から、５１％と算出し、ＩＣＰ－ＭＳ分析による１０Ｂ量の定量により、４９％と算出した。ポリマー１分子に対して約２０個の１０Ｂ－ＢＳＨが導入されたことに相当する。
１Ｈ－ＮＭＲ（４００　ＭＨｚ，Ｄ２Ｏ）δ（ｐｐｍ）：０．５－２．０（ｍ，８２Ｈ），２．０－２．３（ｍ，８２Ｈ），２．８－３．０（ｍ，８２Ｈ），３．７（ｍ，９０９Ｈ），４．２－４．３（ｍ，４１Ｈ）. Polyethylene glycol (Mn = 10,000) -thiolated polyglutamic acid (41mer) -block copolymer (250 mg, 0.014 mmol) and 10 B-BSH (150 mg) synthesized in the same manner as in Synthesis Example 2. , 2 eq) to the thiolated functional group in the block copolymer, 10 mM HEPES (4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid) (pH 8.0) / 500 mM Dissolved in brine (15 ml) and stirred at room temperature for 3 days. The reaction solution was dialyzed 3 times in 150 mM saline, then dialyzed 3 times in deionized water (fractionated molecular weight 3500 Da), and lyophilized to obtain the title compound (266 mg, yield 96%). The introduction rate of 10 B-BSH was determined from the degree of polymerization of boron cluster protons (-B H ; δ (ppm): 0.5-2.0) and polyamino acid by 1 H-NMR analysis (D 2 O). , 51%, and 49% by quantification of the amount of 10 B by ICP-MS analysis. This corresponds to the introduction of about 20 10 B-BSH per molecule of polymer.
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.5-2.0 (m, 82H), 2.0-2.3 (m, 82H), 2.8-3.0 (M, 82H), 3.7 (m, 909H), 4.2-4.3 (m, 41H).
試験例１
ＨＵＶＥＣに対する細胞毒性試験
　実施例１～３及び比較例１及び２の化合物、並びに１０Ｂ－ＢＳＨについて、ＣＣＫ－８アッセイによって、ＨＵＶＥＣ（ヒト臍帯静脈内皮細胞）に対する細胞毒性を調べた。ＨＵＶＥＣを内皮細胞増殖培地キット中に懸濁させ、９６ウェルプレート中に、各ウェルで５０００個の細胞となるように播種し、５％の二酸化炭素存在下、３７℃で２４時間培養した。続いて実施例１～３及び比較例１及び２の化合物、並びに１０Ｂ－ＢＳＨのうち何れかを添加し、各種濃度でさらに７２時間培養した。その後、各ウェルに１０　μＬのｃｅｌｌ　ｃｏｕｎｔｉｎｇ　ｋｉｔ－８溶液を添加し、５％の二酸化炭素存在下、３７℃で２．５時間培養した。最後に、各ウェルについて、マイクロプレートリーダーで４５０　ｎｍの波長吸収を測定した。その結果、図１に示すように、実施例１～３及び比較例１及び２の化合物、並びに１０Ｂ－ＢＳＨについて、ＨＵＶＥＣに対する細胞毒性は認められなかった。 Test example 1
Cytotoxicity test against HUVEC The compounds of Examples 1 to 3 and Comparative Examples 1 and 2 and 10 B-BSH were examined for cytotoxicity against HUVEC (human umbilical vein endothelial cells) by CCK-8 assay. HUVECs were suspended in an endothelial cell growth medium kit, seeded in a 96-well plate so that there were 5000 cells in each well, and cultured at 37 ° C. in the presence of 5% carbon dioxide for 24 hours. Subsequently, any one of the compounds of Examples 1 to 3 and Comparative Examples 1 and 2 and 10 B-BSH was added, and the cells were further cultured at various concentrations for 72 hours. Thereafter, 10 μL of a cell counting kit-8 solution was added to each well, and the cells were cultured at 37 ° C. for 2.5 hours in the presence of 5% carbon dioxide. Finally, 450 nm wavelength absorption was measured for each well with a microplate reader. As a result, as shown in FIG. 1, cytotoxicity against HUVEC was not observed for the compounds of Examples 1 to 3, Comparative Examples 1 and 2, and 10 B-BSH.
試験例２
Ｃ２６がん細胞に対する細胞毒性試験
　実施例１～３及び比較例１及び２の化合物、並びに１０Ｂ－ＢＳＨについて、試験例１の方法と同様にして、Ｃ２６がん細胞に対する細胞毒性を調べた。図２に示すように、実施例１～３及び比較例１及び２の化合物、並びに１０Ｂ－ＢＳＨについて、Ｃ２６がん細胞に対する細胞毒性は認められなかった。 Test example 2
Cytotoxicity test for C26 cancer cells The compounds of Examples 1 to 3 and Comparative Examples 1 and 2, and 10 B-BSH were examined for cytotoxicity to C26 cancer cells in the same manner as in Test Example 1. As shown in FIG. 2, cytotoxicity against C26 cancer cells was not observed for the compounds of Examples 1 to 3, Comparative Examples 1 and 2, and 10 B-BSH.
試験例３
Ｃ２６がん細胞を用いた細胞内取込試験
　実施例１～３及び比較例１及び２の化合物、並びに１０Ｂ－ＢＳＨについて、細胞内取込量を調べた。６ウェルプレートの各ウェルに、１０６個のＣ２６がん細胞を、２　ｍＬのダルベッコ変法イーグル培地（ＤＭＥＭ培地）と共に播種し、２４時間培養した。続いて、実施例１の化合物、実施例２の化合物、実施例３の化合物、比較例１の化合物、比較例２の化合物、１０Ｂ－ＢＳＨを各々、１０Ｂ－ＢＳＨとして１００　μｇ／ｍＬになるように添加した。１、６、２４時間培養後、各細胞をＰＢＳ（リン酸緩衝液）で３回洗浄し、トリプシン処理を行って細胞を回収した。回収した細胞数を数え、９０％硝酸で灰化処理し、最後に１％硝酸で希釈し、ＩＣＰ－ＭＳ分析にてホウ素量を定量した。 Test example 3
Intracellular uptake test using C26 cancer cells The amount of intracellular uptake of the compounds of Examples 1 to 3 and Comparative Examples 1 and 2 and 10 B-BSH was examined. In each well of a 6-well plate, 10 6 C26 cancer cells were seeded with 2 mL of Dulbecco's modified Eagle medium (DMEM medium) and cultured for 24 hours. Subsequently, the compound of Example 1, the compound of Example 2, the compound of Example 3, the compound of Comparative Example 1, the compound of Comparative Example 2, and 10 B-BSH were each converted to 10 B-BSH to 100 μg / mL. It added so that it might become. After culturing for 1, 6, and 24 hours, each cell was washed 3 times with PBS (phosphate buffer), and treated with trypsin to collect the cells. The number of collected cells was counted, ashed with 90% nitric acid, finally diluted with 1% nitric acid, and the amount of boron was quantified by ICP-MS analysis.
　図３に示すように、１０Ｂ－ＢＳＨ単体よりも、実施例１～３及び比較例１及び２の１０Ｂ－ＢＳＨを備えるペプチド化合物の方が、大幅に高い細胞内取込量であった。さらに、ＰＥＧを有さない比較例１及び２の化合物よりも、ＰＥＧを有する実施例１～３の化合物の方が、大幅に高い細胞内取込量であった。 As shown in FIG. 3, the peptide compounds with 10 B-BSH of Examples 1 to 3 and Comparative Examples 1 and 2 had significantly higher intracellular uptake than 10 B-BSH alone. . Furthermore, the amount of cellular uptake of the compounds of Examples 1 to 3 having PEG was significantly higher than the compounds of Comparative Examples 1 and 2 having no PEG.
試験例４
Ｃ２６がん細胞を用いた細胞内取込試験
　実施例１の化合物が細胞内に入ることを確かめた。まず、実施例１の化合物を、Ａｌｅｘａ４８８－ＮＨＳエステルとの縮合によって蛍光標識した。続いて、得られた化合物を、１０Ｂ－ＢＳＨとして２０　μｇ／ｍＬになるようにＣ２６がん細胞に添加した後、継時的に共焦点レーザー顕微鏡（ＣＬＳＭ）にて観察した。図４に示すように、添加後１時間からがん細胞内に入ることを確認した。 Test example 4
Intracellular uptake test using C26 cancer cells It was confirmed that the compound of Example 1 entered the cells. First, the compound of Example 1 was fluorescently labeled by condensation with Alexa488-NHS ester. Subsequently, the obtained compound was added to C26 cancer cells as 10 B-BSH so as to be 20 μg / mL, and subsequently observed with a confocal laser microscope (CLSM). As shown in FIG. 4, it was confirmed that the cancer cells entered from 1 hour after the addition.
試験例５
ＣＴ２６がん細胞を用いた細胞内取込試験
　ホウ素クラスターを、ＰＥＧを有するペプチドに導入することで、がん細胞内により多く取り込まれることを確かめた。ＲＰＭＩ１６４０培地を用いて培養したＣＴ２６がん細胞（マウス大腸がん細胞、ＡＴＣＣ　ＣＲＬ－２６３８）を、２４ウェルプレートの各ウェルに、１０６個ずつ播種した。ホウ素クラスター化合物をペプチド体に導入した実施例７の化合物、或いは合成例２１のホウ素クラスター化合物の対カチオンが２セシウムカチオンである化合物を、３　ｍＭとなるように各ウェルに添加した後、２４時間培養した。培地を除去後、ＰＢＳ（０．５　ｍｌ）で２回洗浄し、トリプシンを用いてプレートからはがした細胞を１４　ｍｌラウンドチューブに回収した。１０％硝酸（０．９　ｍｌ）、６０％硝酸（０．４　ｍｌ）を添加して３０分静置した後、５０℃で３０分、さらに９０℃で２時間加熱して灰化を行った。この溶液をろ過後、脱イオン水で７倍希釈し、ＩＣＰ－ＭＳにて、がん細胞内のホウ素量を定量した。図５に示すように、実施例７の化合物は、合成例２１の化合物の対カチオンが２セシウムカチオンである化合物に対して２倍以上多く、がん細胞内に入ることを確認した。 Test Example 5
Intracellular uptake test using CT26 cancer cells It was confirmed that boron clusters were incorporated more into cancer cells by introducing them into peptides containing PEG. CT26 carcinoma cells (murine colon carcinoma cells, ATCC CRL-2638) cultured with RPMI1640 medium, to each well of 24-well plates, were seeded in 10 6. 24 hours after adding the compound of Example 7 into which the boron cluster compound was introduced into the peptide body or the compound of Example 21 in which the counter cation of the boron cluster compound was a 2 cesium cation to 3 mM. Cultured. After removing the medium, the cells were washed twice with PBS (0.5 ml), and the cells detached from the plate using trypsin were collected in a 14 ml round tube. After adding 10% nitric acid (0.9 ml) and 60% nitric acid (0.4 ml) and allowing to stand for 30 minutes, ashing was carried out by heating at 50 ° C. for 30 minutes and further at 90 ° C. for 2 hours. . This solution was filtered, diluted 7-fold with deionized water, and the amount of boron in cancer cells was quantified by ICP-MS. As shown in FIG. 5, it was confirmed that the compound of Example 7 was more than twice as much as the counter cation of the compound of Synthesis Example 21 compared to the compound of 2 cesium cation and entered the cancer cell.
試験例６
血中滞留性と腫瘍蓄積性
　１０Ｂ－ＢＳＨと、実施例１～３及び比較例１及び２の化合物について、血中滞留性と腫瘍蓄積性を調べた。Ｃ２６がん細胞を皮下移植したＢＡＬＢ／ｃマウスに対し、実施例１～３及び比較例１及び２の化合物及び１０Ｂ－ＢＳＨを、１０Ｂ－ＢＳＨ換算で５０　ｍｇ／ｋｇ量となるよう、各化合物の生理食塩水溶液０．２　ｍＬ量を静脈内投与し、血液と腫瘍を経時的に採取した。血液はヘパリン処理し、遠心分離で血漿を取得した。血漿、腫瘍のサンプルは塩酸／６０％硝酸（３／１）混合液、或いは９０％硝酸水溶液で処理して組織を分解し、１％硝酸で希釈し、ろ過を行った後、ＩＣＰ－ＭＳにて１０Ｂ量を測定した。 Test Example 6
Blood retention and tumor accumulation 10 B-BSH and the compounds of Examples 1 to 3 and Comparative Examples 1 and 2 were examined for blood retention and tumor accumulation. For BALB / c mice transplanted subcutaneously with C26 cancer cells, the compounds of Examples 1 to 3 and Comparative Examples 1 and 2 and 10 B-BSH were adjusted to a dose of 50 mg / kg in terms of 10 B-BSH. A 0.2 mL volume of physiological saline solution of each compound was intravenously administered, and blood and tumor were collected over time. The blood was heparinized and plasma was obtained by centrifugation. Plasma and tumor samples were treated with hydrochloric acid / 60% nitric acid (3/1) mixed solution or 90% aqueous nitric acid solution to decompose the tissue, diluted with 1% nitric acid, filtered, and then ICP-MS 10 B amount was measured.
　図６に示すように、実施例１～３及び比較例１及び２の化合物は、１０Ｂ－ＢＳＨ単体よりも、長い血中滞留性を示し、さらに、ＰＥＧを有する実施例１～３の化合物は、ＰＥＧを有さない比較例１及び２の化合物よりも、長い血中滞留性を示した。また、図７に示すように、実施例１～３及び比較例１及び２の化合物は、１０Ｂ－ＢＳＨ単体よりも、６～１０倍多く１０Ｂ－ＢＳＨを腫瘍に集積させ、さらに、ＰＥＧを有する実施例１～３の化合物は、ＰＥＧを有さない比較例１及び２の化合物よりも、多く１０Ｂ－ＢＳＨを腫瘍に集積させ、高い集積量を保持する時間も長いことがわかった。 As shown in FIG. 6, the compounds of Examples 1 to 3 and Comparative Examples 1 and 2 showed longer retention in blood than 10 B-BSH alone, and the compounds of Examples 1 to 3 having PEG. Showed longer retention in blood than the compounds of Comparative Examples 1 and 2 without PEG. In addition, as shown in FIG. 7, the compounds of Examples 1 to 3 and Comparative Examples 1 and 2 accumulate 10 B-BSH in the tumor 6 to 10 times more than 10 B-BSH alone, and further PEG It was found that the compounds of Examples 1 to 3 having a higher concentration of 10 B-BSH accumulated in the tumor and a longer amount of accumulation than the compounds of Comparative Examples 1 and 2 having no PEG. .
試験例７
腫瘍蓄積性
　天然型Ｂ－ＢＳＨと、実施例７の化合物について、各化合物の生理食塩水溶液をマウスに静脈内投与し、腫瘍蓄積性を調べた。ＣＴ２６がん細胞を皮下移植したＢＡＬＢ／ｃマウスに対し、実施例７の化合物を天然型Ｂ－ＢＳＨ換算で１０　ｍｇ／ｋｇ及び３０　ｍｇ／ｋｇ量となるように、また天然型Ｂ－ＢＳＨを１００　ｍｇ／ｋｇ量となるように、各々０．２　ｍＬ量を静脈内投与し、腫瘍を経時的に採取した。採取した腫瘍のサンプルを６０％硝酸水溶液中、５０℃で１時間、続いて９０℃で２時間加熱し、組織を分解した。ろ過を行った後、純水で２０倍希釈し、ＩＣＰ－ＭＳにて天然型Ｂ量を測定した。 Test Example 7
Tumor Accumulation Regarding natural B-BSH and the compound of Example 7, physiological saline solution of each compound was intravenously administered to mice, and tumor accumulation was examined. For BALB / c mice transplanted subcutaneously with CT26 cancer cells, the amount of the compound of Example 7 was 10 mg / kg and 30 mg / kg in terms of natural B-BSH, and natural B-BSH was added. Each 0.2 mL amount was intravenously administered so that the amount was 100 mg / kg, and tumors were collected over time. The collected tumor samples were heated in a 60% aqueous nitric acid solution at 50 ° C. for 1 hour and then at 90 ° C. for 2 hours to decompose the tissue. After filtration, it was diluted 20 times with pure water, and the amount of natural B was measured by ICP-MS.
　図８に示すように、実施例７の化合物は、天然型Ｂ－ＢＳＨ単体よりも、３～５倍、さらには時間経過と共にそれ以上多くホウ素クラスターを腫瘍に集積させ、高い集積量を保持する時間も長いことがわかった。 As shown in FIG. 8, the compound of Example 7 accumulates boron clusters in the tumor 3 to 5 times more than natural B-BSH alone, and more over time, and maintains a high accumulation amount. I found that the time was long.
試験例８
抗がん効果
　Ｃ２６がん細胞を皮下移植したＢＡＬＢ／ｃマウスに対して、実施例１の化合物、或いは１０Ｂ－ＢＳＨを、１０Ｂ－ＢＳＨ換算で１００　ｍｇ／ｋｇとなるよう、各化合物の生理食塩水溶液０．２　ｍＬ量を静脈内投与した。２４時間後、このマウスに対して、１．６－２．２ｘ１０１２　ｎｅｕｔｒｏｎ／ｃｍ２の量の熱中性子線を１時間局所照射し、抗がん効果について調べた。熱中性子線照射前後の腫瘍の大きさ（Ｖ）は、Ｖ＝（ａｘｂ２）／２の式にて算出した（ａ、ｂは、各々腫瘍をキャリパーで測定した時の長径、短径）。コントロールとして、Ｃ２６がん細胞を皮下移植したＢＡＬＢ／ｃマウスに対して、実施例１の化合物、或いは１０Ｂ－ＢＳＨを、１０Ｂ－ＢＳＨ換算で１００　ｍｇ／ｋｇとなるよう、０．２　ｍＬ量を静脈内投与し、熱中性子線を照射せずに、同様に腫瘍の大きさを算出した。また、もう一つのコントロールとして、Ｃ２６がん細胞を皮下移植したＢＡＬＢ／ｃマウスに対して、ＰＢＳ（リン酸緩衝液）を投与して、熱中性子線を照射、或いは照射せずに、同様に腫瘍の大きさを算出した。 Test Example 8
Anti-cancer effect For BALB / c mice transplanted subcutaneously with C26 cancer cells, the compound of Example 1 or 10 B-BSH was added to each compound so as to be 100 mg / kg in terms of 10 B-BSH. A physiological saline solution (0.2 mL) was intravenously administered. After 24 hours, the mice were locally irradiated with a thermal neutron beam in an amount of 1.6-2.2 × 10 12 neutron / cm 2 for 1 hour to examine the anticancer effect. The size (V) of the tumor before and after thermal neutron irradiation was calculated by the equation V = (axb 2 ) / 2 (a and b are the major axis and minor axis when the tumor was measured with a caliper, respectively). As a control, BALB / c mice transplanted subcutaneously with C26 cancer cells were treated with 0.2 mL of the compound of Example 1 or 10 B-BSH so that the concentration was 100 mg / kg in terms of 10 B-BSH. The amount of the tumor was administered intravenously, and the size of the tumor was calculated in the same manner without irradiation with thermal neutrons. Moreover, as another control, PBS (phosphate buffer solution) was administered to BALB / c mice transplanted subcutaneously with C26 cancer cells, and irradiated with or without thermal neutron radiation. Tumor size was calculated.
　図９に示すように、実施例１の化合物を投与後、熱中性子線を照射した場合、腫瘍増殖は大きく抑制され、腫瘍はほとんど大きくならなかった。一方、１０Ｂ－ＢＳＨ単体或いはリン酸緩衝液を投与後、熱中性子線を照射した場合、実施例１の化合物、１０Ｂ－ＢＳＨ単体、或いはリン酸緩衝液を投与後、熱中性子線を照射しない場合のコントロール群と同様に、速い速度で腫瘍が増殖した。このことから、本発明の化合物は、ホウ素中性子捕捉療法によって、高い抗腫瘍効果を有することがわかった。 As shown in FIG. 9, when the compound of Example 1 was administered and irradiated with a thermal neutron beam, tumor growth was greatly suppressed, and the tumor was hardly enlarged. On the other hand, when irradiated with thermal neutrons after administration of 10 B-BSH alone or phosphate buffer, it is irradiated with thermal neutrons after administration of the compound of Example 1, 10 B-BSH alone or phosphate buffer. Similar to the control group without, the tumor grew at a fast rate. From this, it was found that the compound of the present invention has a high antitumor effect by boron neutron capture therapy.
　さらに、マウスの全身毒性の指標として、本実験で用いたマウスの体重を測定したところ、図１０に示すように、実施例１の化合物、或いは１０Ｂ－ＢＳＨ単体を静脈内投与したマウス、熱中性子線を照射したマウス、照射しなかったマウス、全てにおいて、明らかな体重抑制は見られず、本発明の化合物、及び本発明の化合物を用いたホウ素中性子捕捉療法は高い安全性を有することがわかった。 Further, the body weight of the mouse used in this experiment was measured as an indicator of the systemic toxicity of the mouse. As shown in FIG. 10, the mouse of Example 1 or 10 B-BSH alone administered intravenously, In mice that were irradiated with neutrons and mice that were not irradiated, there was no apparent weight suppression, and the compounds of the present invention and boron neutron capture therapy using the compounds of the present invention have high safety. all right.
試験例９
腫瘍浸透性
　実施例１の化合物と、がん治療薬として認可されている抗がん剤ドキソルビシンを担持したリポソーム製剤Ｄｏｘｉｌ（登録商標）について、担がんマウスを用いた腫瘍浸透性の比較を調べた。ヒト膵臓腺がんＢｘＰＣ３細胞、或いはＣ２６がん細胞を皮下移植したＢＡＬＢ／ｃヌードマウスに対して、１０Ｂ換算で１０　ｍｇ／ｋｇ量の、実施例１の化合物をＡｌｅｘａ６４７（登録商標）で蛍光標識した化合物と、ドキソルビシン換算で１０　ｍｇ／ｋｇ量のＤｏｘｉｌ（登録商標）の両方を、静脈内投与した。投与して２４時間後、腫瘍を切除し、サンプリングして、ミクロトームにて１０　μｍの薄さに切った。腫瘍切片について、共焦点走査型顕微鏡ＬＳＭ７８０で画像観察し、画像はバックグラウンドの蛍光強度と同じように標準化した。腫瘍は、ヘマトキシリン－エオシン染色のため、同様に５　μｍの薄さに切り、一体型蛍光顕微鏡ＢＺ－Ｘ７００で画像観察した。 Test Example 9
Tumor penetrability A comparison of tumor penetrability using tumor-bearing mice was examined for the liposome preparation Doxil (registered trademark) carrying the compound of Example 1 and the anticancer drug doxorubicin approved as a cancer therapeutic agent. It was. Human pancreatic adenocarcinoma BxPC3 cells, or C26 cancer cell to BALB / c nude mice were implanted subcutaneously, fluorescence 10 mg / kg weight at 10 B terms, the compound of Example 1 with Alexa647 (R) Both the labeled compound and 10 mg / kg Doxil® in terms of doxorubicin were administered intravenously. Twenty-four hours after dosing, tumors were excised, sampled and cut to a thickness of 10 μm with a microtome. The tumor sections were imaged with a confocal scanning microscope LSM780, and the images were normalized in the same way as the background fluorescence intensity. The tumor was similarly cut to a thickness of 5 μm for hematoxylin-eosin staining, and images were observed with an integrated fluorescence microscope BZ-X700.
　図１１に示すように、実施例１の化合物をＡｌｅｘａ６４７（登録商標）で蛍光標識した化合物とＤｏｘｉｌ（登録商標）は、共に多血性のＣ２６腫瘍に同様に分布した。一方、乏血性のＢｘＰＣ３腫瘍モデルには、Ｄｏｘｉｌ（登録商標）はほとんど分布していないのに対し、実施例１の化合物をＡｌｅｘａ６４７（登録商標）で蛍光標識した化合物は、腫瘍内に均一に分布した。この結果から、本発明の化合物は、ナノ粒子より腫瘍への浸透性が高く、がん細胞全体にホウ素を送達できることがわかった。 As shown in FIG. 11, both the compound obtained by fluorescently labeling the compound of Example 1 with Alexa 647 (registered trademark) and Doxil (registered trademark) were similarly distributed in the polycytic C26 tumor. On the other hand, in the ischemic BxPC3 tumor model, Doxil (registered trademark) is hardly distributed, whereas the compound of Example 1 fluorescently labeled with Alexa 647 (registered trademark) is uniformly distributed in the tumor. did. From these results, it was found that the compound of the present invention has higher tumor penetration than nanoparticles and can deliver boron to the entire cancer cells.
　本発明の化合物は、ＥＰＲ効果による優れたがん集積性、及びナノ粒子化しないことによる腫瘍組織内での高い浸透性を示し、がん細胞内に効率的に取り込まれるため、ホウ素中性子捕捉療法における医薬として有用である。 Since the compound of the present invention exhibits excellent cancer accumulation due to the EPR effect and high permeability in tumor tissue due to non-nanoparticulation, it is efficiently taken up into cancer cells, so boron neutron capture therapy It is useful as a pharmaceutical.
　本出願は、日本国で２０１６年３月２３日に出願された特願２０１６－０５９２２５号を基礎としており、その内容は本明細書にすべて包含されるものである。
  This application is based on Japanese Patent Application No. 2016-059225 filed on March 23, 2016 in Japan, the contents of which are incorporated in full herein.

Claims (19)

  1. 　ホウ素クラスター及びポリエーテルを有し、中性アミノ酸及び酸性アミノ酸から選ばれるアミノ酸を構成アミノ酸とするペプチド化合物又はその塩。 A peptide compound having a boron cluster and a polyether, and an amino acid selected from neutral amino acids and acidic amino acids, or a salt thereof.
  2. 　ホウ素クラスターが側鎖に結合したアミノ酸残基を少なくとも１個有する請求項１に記載のペプチド化合物又はその塩。 The peptide compound or a salt thereof according to claim 1, wherein the boron cluster has at least one amino acid residue bonded to the side chain.
  3. 　ホウ素クラスターが側鎖に結合したアミノ酸残基が、式（１）：
    Figure JPOXMLDOC01-appb-C000001
    ［式中、Ｒ１は、それぞれ独立して、ホウ素クラスターからなる１価の基又はそれがリンカー構造を介して結合する１価の基で置換された、中性アミノ酸及び酸性アミノ酸から選ばれるα－アミノ酸の側鎖を示し、当該α－アミノ酸の側鎖は、それが結合する炭素原子及び該炭素原子に隣接する窒素原子と一緒になって、環を形成していてもよく、環を形成する場合は該窒素原子に結合するＨは存在しない。］
    で表される、請求項２に記載のペプチド化合物又はその塩。     An amino acid residue having a boron cluster bonded to a side chain is represented by the formula (1):
    Figure JPOXMLDOC01-appb-C000001
    [Wherein, each R 1 independently represents an α selected from a neutral amino acid and an acidic amino acid substituted with a monovalent group consisting of a boron cluster or a monovalent group to which it is bonded via a linker structure. -Indicates the side chain of an amino acid, and the side chain of the α-amino acid may form a ring together with the carbon atom to which it is attached and the nitrogen atom adjacent to the carbon atom. When it does, H couple | bonded with this nitrogen atom does not exist. ]
    The peptide compound or its salt of Claim 2 represented by these.
  4. 　Ｒ１が、それぞれ独立して、式（Ｂ１）：
    Figure JPOXMLDOC01-appb-C000002
     
    ［式中、Ｍ２＋は、カチオンを示し、●は、ＢＨを示し、＊は、リンカー構造との結合部位を示す。］
    で表される基がリンカー構造を介して結合する１価の基で置換された、中性アミノ酸及び酸性アミノ酸から選ばれるα－アミノ酸の側鎖である、請求項３に記載のペプチド化合物又はその塩。     Each R 1 independently represents the formula (B1):
    Figure JPOXMLDOC01-appb-C000002

    [ Wherein M 2+ represents a cation, ● represents BH, and * represents a binding site with a linker structure. ]
    The peptide compound according to claim 3, which is a side chain of an α-amino acid selected from a neutral amino acid and an acidic amino acid, wherein the group represented by is substituted with a monovalent group bonded through a linker structure salt.
  5. 　Ｒ１が、それぞれ独立して、式（Ｒ１ａ）：
    Figure JPOXMLDOC01-appb-C000003
    ［式中、Ｒａは、それぞれ独立して、水素原子、炭素数が１～６のアルキル基で置換されていてもよいカルボキシ基、炭素数が１～６のアルキル基で置換されていてもよいヒドロキシ基、炭素数が１～６のアルキル基で置換されていてもよいメルカプト基、炭素数が６～１４のアリール基、又は炭素数が１～６のアルキル基を示し；Ｑ２は、それぞれ独立して、－Ｏ－、－Ｓ－、－ＮＨ－、－ＳＯ２－、－ＣＯ－、－ＮＨＣＯ－、－ＣＯＮＨ－、－ＯＣＯ－、－ＣＯＯ－、－ＳＣＯ－、－ＣＯＳ－、－ＮＨＣＯＮＨ－、－ＮＨＣＳＮＨ－、－ＯＣＯＮＨ－、－ＮＨＣＯＯ－、－ＳＳ－、－Ｐｈ－、－ＯＰｈＯ－、－ＯＰｈ－、－ＰｈＯ－、－ＳＰｈＳ－、－ＳＰｈ－、－ＰｈＳ－、－ＮＨＰｈＮＨ－、－ＮＨＰｈ－、－ＰｈＮＨ－、－複素環－、－Ｏ－複素環－Ｏ－、－Ｏ－複素環－、－複素環－Ｏ－、－Ｓ－複素環－Ｓ－、－Ｓ－複素環－、－複素環－Ｓ－、－ＮＨ－複素環－ＮＨ－、－ＮＨ－複素環－又は－複素環－ＮＨ－（ここで、Ｐｈは、１，２－フェニレン、１，３－フェニレン又は１，４－フェニレンを示す。）を示し；Ｐ３は、－Ｏ－、－ＮＨ－又は－Ｓ－を示し；Ｍ２＋は、カチオンを示し；●は、ＢＨを示し；ｄは、１又は２を示し；ｆは、それぞれ独立して、１～２０の整数を示し；ｇは、０～２０の整数を示し；＊＊は、α－アミノ酸のα－炭素との結合部位を示す。］
    で表される基である、請求項４に記載のペプチド化合物又はその塩。     Each R 1 independently represents the formula (R1a):
    Figure JPOXMLDOC01-appb-C000003
    [Wherein, each R a is independently a hydrogen atom, a carboxy group optionally substituted with an alkyl group having 1 to 6 carbon atoms, or an alkyl group having 1 to 6 carbon atoms. good hydroxy group, an alkyl mercapto group optionally substituted with a group with a carbon number of 1-6, an aryl group, or an alkyl group having 1 to 6 carbon atoms having a carbon number of 6 ~ 14; Q 2 is Independently, —O—, —S—, —NH—, —SO 2 —, —CO—, —NHCO—, —CONH—, —OCO—, —COO—, —SCO—, —COS—, —NHCONH—, —NHCSNH—, —OCONH—, —NHCOO—, —SS—, —Ph—, —OPhO—, —OPh—, —PhO—, —SPhS—, —SPh—, —PhS—, —NHPhNH -, -NHPh-, -PhNH-, -complex Ring-, -O-heterocycle-O-, -O-heterocycle-, -heterocycle-O-, -S-heterocycle-S-, -S-heterocycle-, -heterocycle-S-,- NH-heterocycle-NH-, -NH-heterocycle- or -heterocycle-NH- (wherein Ph represents 1,2-phenylene, 1,3-phenylene or 1,4-phenylene). P 3 represents —O—, —NH— or —S—; M 2+ represents a cation; ● represents BH; d represents 1 or 2; f represents each independently G represents an integer of 0 to 20; ** represents a binding site with an α-carbon of an α-amino acid. ]
    The peptide compound or its salt of Claim 4 which is group represented by these.
  6. 　Ｒ１のα－アミノ酸が、グリシン、アラニン、バリン、ロイシン、イソロイシン、セリン、トレオニン、システイン、メチオニン、アスパラギン、グルタミン、フェニルアラニン、チロシン、アスパラギン酸、グルタミン酸、ホモセリン、ノルロイシン、ノルバリン、チロニン、シトルリン、α-アミノ酪酸、ホモシステイン及びペニシラミンからなる群から
    選ばれる、請求項３～５の何れか１項に記載のペプチド化合物又はその塩。     The α-amino acid of R 1 is glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, asparagine, glutamine, phenylalanine, tyrosine, aspartic acid, glutamic acid, homoserine, norleucine, norvaline, thyronine, citrulline, α 6. The peptide compound or a salt thereof according to any one of claims 3 to 5, which is selected from the group consisting of aminobutyric acid, homocysteine and penicillamine.
  7. 　Ｒ１のα－アミノ酸が、グルタミン酸及びアスパラギン酸からなる群から選ばれる、請求項３～５の何れか１項に記載のペプチド化合物又はその塩。 The peptide compound or a salt thereof according to any one of claims 3 to 5, wherein the α-amino acid of R 1 is selected from the group consisting of glutamic acid and aspartic acid.
  8. 　ペプチド化合物のＣ末端のカルボキシ基及び／又はペプチド化合物のＮ末端のアミノ基がポリエーテルからなる１価の基で置換されている、請求項１～７の何れか１項に記載のペプチド化合物又はその塩。 The peptide compound according to any one of claims 1 to 7, wherein the C-terminal carboxy group of the peptide compound and / or the N-terminal amino group of the peptide compound is substituted with a monovalent group comprising a polyether. Its salt.
  9. 　ペプチド化合物のＣ末端のカルボキシ基及び／又はペプチド化合物のＮ末端のアミノ基が、式（Ｐ１）：
    Figure JPOXMLDOC01-appb-C000004
    ［式中、Ｚは、置換対象がカルボキシ基の場合－Ｏ－、－ＮＨ－又は－Ｓ－を、置換対象がアミノ基の場合－ＣＯ－、－ＣＯＮＨ－又は－ＣＯＯ－を示し、Ｒｄ及びＲｅは、それぞれ独立して水素原子又は炭素数が１～６のアルキル基を示し、ｓは、それぞれ独立して１～１０の整数であり、ｔは、それぞれ独立して２～１０の整数であり、ｕは、それぞれ独立して１以上の整数であり、＊＊＊は、結合部位を示す。］
    で表される基で置換されている、請求項１～８の何れか１項に記載のペプチド化合物又はその塩。     The C-terminal carboxy group of the peptide compound and / or the N-terminal amino group of the peptide compound is represented by the formula (P1):
    Figure JPOXMLDOC01-appb-C000004
    [In the formula, Z represents —O—, —NH— or —S— when the substitution target is a carboxy group, and —CO—, —CONH— or —COO— when the substitution target is an amino group, R d And R e each independently represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, s is each independently an integer of 1 to 10, and t is independently 2 to 10 Each is an integer, u is each independently an integer of 1 or more, and *** represents a binding site. ]
    The peptide compound or a salt thereof according to any one of claims 1 to 8, which is substituted with a group represented by:
  10. 　ペプチドがポリアミノ酸である請求項１～９の何れか１項に記載のペプチド化合物又はその塩。 The peptide compound or a salt thereof according to any one of claims 1 to 9, wherein the peptide is a polyamino acid.
  11. 　ペプチドのアミノ酸残基数に対するホウ素クラスターの結合数の比が０．１以上である、請求項１～１０の何れか１項に記載のペプチド化合物又はその塩。 The peptide compound or salt thereof according to any one of claims 1 to 10, wherein the ratio of the number of bonds of boron clusters to the number of amino acid residues of the peptide is 0.1 or more.
  12. 　ペプチドのアミノ酸残基数が２００以下である、請求項１～１１の何れか１項に記載のペプチド化合物又はその塩。 The peptide compound or salt thereof according to any one of claims 1 to 11, wherein the peptide has 200 or less amino acid residues.
  13. 　ポリエーテルの数平均分子量が、５０，０００未満である、請求項１～１２の何れか１項に記載のペプチド化合物又はその塩。 The peptide compound or salt thereof according to any one of claims 1 to 12, wherein the polyether has a number average molecular weight of less than 50,000.
  14. 　分子量が、１００，０００未満である、請求項１～１３の何れか１項に記載のペプチド化合物又はその塩。 The peptide compound or a salt thereof according to any one of claims 1 to 13, having a molecular weight of less than 100,000.
  15. 　式（Ｉａ）又は式（Ｉｂ）：
    Figure JPOXMLDOC01-appb-C000005
    ［式中、
    　Ｒ１は、それぞれ独立して、ホウ素クラスターからなる１価の基又はそれがリンカー構造を介して結合する１価の基で置換された、中性アミノ酸及び酸性アミノ酸から選ばれるα－アミノ酸の側鎖を示し、
    　Ｒ２は、それぞれ独立して、置換基を有していてもよい、中性アミノ酸及び酸性アミノ酸から選ばれるα－アミノ酸の側鎖を示し、
    　Ｒ１及びＲ２のα－アミノ酸の側鎖は、それぞれ、それが結合する炭素原子及び該炭素原子に隣接する窒素原子と一緒になって、環を形成していてもよく、環を形成する場合は該窒素原子に結合するＨは存在せず、
    　ｂが１以上の場合、Ｒ１を有するα－アミノ酸残基とＲ２を有するα－アミノ酸残基とは、任意に共重合しており、
    　Ｘ１、及びｃが１の場合のＹ２は、それぞれ独立して、
    （１）水素原子、
    （２）炭素数が１～６のアルキル基、
    （３）炭素数が２～７のアルキルカルボニル基、又は
    （４）ポリエーテルからなる１価の基を示し、
    　Ｘ２、及びｃが１の場合のＹ１は、それぞれ独立して、
    （１）炭素数が１～６のアルキル基で置換されていてもよいヒドロキシ基、
    （２）炭素数が１～６のアルキル基で置換されていてもよいアミノ基、又は
    （３）ポリエーテルからなる１価の基を示し、
    　ｃが２以上の場合のＹ１及びＹ２は、それぞれ、ポリエーテルからなるｃ価の基を示し、
    　式（Ｉａ）においては、Ｒ２のα－アミノ酸の側鎖の置換基、並びにＸ１及びＹ１のうち少なくともいずれか１つが、式（Ｉｂ）においては、Ｒ２のα－アミノ酸の側鎖の置換基、並びにＸ２及びＹ２のうち少なくともいずれか１つが、ポリエーテルからなる１価の基又はポリエーテルからなるｃ価の基であり、
    　ａ及びｃは、それぞれ独立して、１以上の整数を示し、
    　ｂは、０以上の整数を示し、
    　ａとｂの合計は、２以上である。］
    で表される、請求項１～１４の何れか１項に記載のペプチド化合物又はその塩。     Formula (Ia) or Formula (Ib):
    Figure JPOXMLDOC01-appb-C000005
    [Where:
    R 1 is each independently an α-amino acid selected from a neutral amino acid and an acidic amino acid substituted with a monovalent group consisting of a boron cluster or a monovalent group to which it is bonded via a linker structure Showing the chain
    R 2 each independently represents a side chain of an α-amino acid selected from a neutral amino acid and an acidic amino acid, which may have a substituent,
    The side chain of the α-amino acid of R 1 and R 2 may be combined with the carbon atom to which it is attached and the nitrogen atom adjacent to the carbon atom to form a ring. In this case, there is no H bonded to the nitrogen atom,
    When b is 1 or more, the α-amino acid residue having R 1 and the α-amino acid residue having R 2 are optionally copolymerized,
    X 1 and Y 2 when c is 1 are each independently
    (1) a hydrogen atom,
    (2) an alkyl group having 1 to 6 carbon atoms,
    (3) an alkylcarbonyl group having 2 to 7 carbon atoms, or (4) a monovalent group comprising a polyether,
    X 2 and Y 1 when c is 1 are each independently
    (1) a hydroxy group which may be substituted with an alkyl group having 1 to 6 carbon atoms,
    (2) an amino group which may be substituted with an alkyl group having 1 to 6 carbon atoms, or (3) a monovalent group comprising a polyether,
    Y 1 and Y 2 in the case where c is 2 or more each represent a c-valent group composed of a polyether;
    In formula (Ia), at least one of the substituents on the side chain of the α-amino acid of R 2 and X 1 and Y 1 is the side chain of the α-amino acid of R 2 in formula (Ib) And at least one of X 2 and Y 2 is a monovalent group made of a polyether or a c-valent group made of a polyether,
    a and c each independently represent an integer of 1 or more,
    b represents an integer of 0 or more,
    The sum of a and b is 2 or more. ]
    The peptide compound or a salt thereof according to any one of claims 1 to 14, which is represented by:
  16. 　ｃが１である、請求項１４又は１５に記載のペプチド化合物又はその塩。 The peptide compound or a salt thereof according to claim 14 or 15, wherein c is 1.
  17. 　腫瘍疾患のホウ素中性子捕捉療法に用いるための請求項１～１６の何れか１項に記載のペプチド化合物又はその塩。 The peptide compound or salt thereof according to any one of claims 1 to 16, for use in boron neutron capture therapy for tumor diseases.
  18. 　請求項１～１６の何れか１項に記載のペプチド化合物又はその塩、及び薬学的に許容される担体を含む医薬組成物。 A pharmaceutical composition comprising the peptide compound or a salt thereof according to any one of claims 1 to 16, and a pharmaceutically acceptable carrier.
  19. 　腫瘍疾患のホウ素中性子捕捉療法に用いるための医薬を製造するための請求項１～１６の何れか１項に記載のペプチド化合物又はその塩の使用。 Use of the peptide compound or a salt thereof according to any one of claims 1 to 16 for producing a medicament for use in boron neutron capture therapy for tumor diseases.
PCT/JP2017/011843 2016-03-23 2017-03-23 Boron cluster bonded peptide compound WO2017164334A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2018507416A JPWO2017164334A1 (en) 2016-03-23 2017-03-23 Boron cluster-binding peptide compound

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2016059225 2016-03-23
JP2016-059225 2016-03-23

Publications (1)

Publication Number Publication Date
WO2017164334A1 true WO2017164334A1 (en) 2017-09-28

Family

ID=59900523

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2017/011843 WO2017164334A1 (en) 2016-03-23 2017-03-23 Boron cluster bonded peptide compound

Country Status (2)

Country Link
JP (1) JPWO2017164334A1 (en)
WO (1) WO2017164334A1 (en)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018097335A1 (en) * 2016-11-25 2018-05-31 国立大学法人 岡山大学 Novel bsh complex for boron neutron capture therapy
WO2019160129A1 (en) * 2018-02-15 2019-08-22 国立大学法人 岡山大学 Cancer lesion tissue evaluation for optimizing effect of boron neutron capture therapy
WO2020040296A1 (en) * 2018-08-23 2020-02-27 森田薬品工業株式会社 Pegylated boron cluster compound, antitumor agent including pegylated boron cluster compound, and sensitizer including pegylated boron cluster
US11324703B2 (en) 2017-12-15 2022-05-10 3-D Matrix, Ltd. Surfactant peptide nanostructures and uses thereof in drug delivery
JP7453658B2 (en) 2019-01-22 2024-03-21 学校法人近畿大学 Boron-containing compounds and drugs containing them

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006027028A1 (en) * 2004-09-10 2006-03-16 L. Molteni & C. Dei Fratelli Alitti Societa' Di Esercizio S.P.A. Boronated metal-phthalocyanines, process for their preparation, pharmaceutical compositions comprising them and use thereof
WO2009116509A1 (en) * 2008-03-18 2009-09-24 日本化薬株式会社 Polymer conjugate of physiologically active substance
WO2012133884A1 (en) * 2011-03-31 2012-10-04 ナノキャリア株式会社 Pharmaceutical composition containing block copolymer comprising boric acid compound
JP2015044920A (en) * 2013-08-27 2015-03-12 国立大学法人 筑波大学 Polymeric boron compound and its use
WO2017026276A1 (en) * 2015-08-07 2017-02-16 国立大学法人東京工業大学 Pharmaceutical composition containing conjugate of boron-containing compound and protein

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006027028A1 (en) * 2004-09-10 2006-03-16 L. Molteni & C. Dei Fratelli Alitti Societa' Di Esercizio S.P.A. Boronated metal-phthalocyanines, process for their preparation, pharmaceutical compositions comprising them and use thereof
WO2009116509A1 (en) * 2008-03-18 2009-09-24 日本化薬株式会社 Polymer conjugate of physiologically active substance
WO2012133884A1 (en) * 2011-03-31 2012-10-04 ナノキャリア株式会社 Pharmaceutical composition containing block copolymer comprising boric acid compound
JP2015044920A (en) * 2013-08-27 2015-03-12 国立大学法人 筑波大学 Polymeric boron compound and its use
WO2017026276A1 (en) * 2015-08-07 2017-02-16 国立大学法人東京工業大学 Pharmaceutical composition containing conjugate of boron-containing compound and protein

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
MI, PENG ET AL.: "Block copolymer-boron cluster conjugate for effective boron neutron capture therapy of solid tumors", JOURNAL OF CONTROLLED RELEASE, vol. 254, 21 March 2017 (2017-03-21), pages 1 - 9, XP085063387, DOI: doi:10.1016/j.jconrel.2017.03.036 *
TAKESHI NAGASAKI ET AL.: "Houso cluster shushoku polyamine kara naru nano ryushi no sakusei to shuyo shusekisei", POLYMER PREPRINTS, vol. 58, no. 2, 2009 *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018097335A1 (en) * 2016-11-25 2018-05-31 国立大学法人 岡山大学 Novel bsh complex for boron neutron capture therapy
US11324703B2 (en) 2017-12-15 2022-05-10 3-D Matrix, Ltd. Surfactant peptide nanostructures and uses thereof in drug delivery
WO2019160129A1 (en) * 2018-02-15 2019-08-22 国立大学法人 岡山大学 Cancer lesion tissue evaluation for optimizing effect of boron neutron capture therapy
CN111971563A (en) * 2018-02-15 2020-11-20 立美基股份有限公司 Cancer lesion tissue assessment to optimize the effects of boron neutron capture therapy
JPWO2019160129A1 (en) * 2018-02-15 2021-03-04 国立大学法人 岡山大学 Cancer lesion tissue evaluation that optimizes the effect of boron neutron capture therapy
JP7221539B2 (en) 2018-02-15 2023-02-14 国立大学法人 岡山大学 Cancer lesion tissue evaluation to optimize the effect of boron neutron capture therapy
CN111971563B (en) * 2018-02-15 2023-09-08 立美基股份有限公司 Cancerous tissue assessment for optimizing the effects of boron neutron capture therapy
WO2020040296A1 (en) * 2018-08-23 2020-02-27 森田薬品工業株式会社 Pegylated boron cluster compound, antitumor agent including pegylated boron cluster compound, and sensitizer including pegylated boron cluster
JPWO2020040296A1 (en) * 2018-08-23 2021-08-26 森田薬品工業株式会社 An anti-tumor agent containing a PEGylated boron cluster compound and a PEGylated boron cluster compound, and a sensitizer containing a PEGylated boron cluster
JP7337342B2 (en) 2018-08-23 2023-09-04 森田薬品工業株式会社 Pegylated Boron Cluster Compounds and Antitumor Agents Comprising Pegylated Boron Cluster Compounds and Sensitizers Comprising Pegylated Boron Clusters
JP7453658B2 (en) 2019-01-22 2024-03-21 学校法人近畿大学 Boron-containing compounds and drugs containing them

Also Published As

Publication number Publication date
JPWO2017164334A1 (en) 2019-02-28

Similar Documents

Publication Publication Date Title
WO2017164334A1 (en) Boron cluster bonded peptide compound
JP4717016B2 (en) O, O'-amidmalonate and N, O-amidmalonate platinum complexes
CN111542344A (en) Ligand-drug-conjugates comprising a single molecular weight polymyosine
EP2090607B1 (en) Polymeric derivative of nucleic acid metabolic antagonist
US20090253651A1 (en) Drug delivery system based on regioselectively amidated hyaluronic acid
AU2011212742B2 (en) Polyanionic multivalent macromolecules for intracellular targeting of proliferation and protein synthesis
AU2010254013A1 (en) Polyal drug conjugates comprising variable rate-releasing linkers
Yu et al. Fabrication and evaluation of reduction-sensitive supramolecular hydrogel based on cyclodextrin/polymer inclusion for injectable drug-carrier application
CA2636599A1 (en) Modified macromolecule
JP2010533202A (en) Polymeric drug delivery system comprising a polysubstituted aromatic moiety
KR20090057235A (en) Lysine-based polymeric linkers
CA2257233A1 (en) Process for producing drug complexes
KR20100051722A (en) Polymeric linkers containing pyridyl disulfide moieties
BRPI0721759A2 (en) "AMPHIPHYLIC self-assembling polymers as anticancer agents"
JP2010533233A (en) Polymeric drug delivery system containing aromatic allylic acid
WO2013060919A1 (en) Controlled synthesis of polyglutamates with low polydispersity and versatile architectures
WO2015035446A1 (en) Macromolecules of dendrimer-platinum conjugates
EP2613810B1 (en) Biodegradable, water soluble and ph responsive poly(organo)phosphazenes
WO2014084378A1 (en) Anticancer drug comprising cyclic rgd sequence-containing peptide
EP4028447B1 (en) Polymer and composition from renewable source
US20060104908A1 (en) Polymerization method for the synthesis of polypeptide imaging agents
WO2006124205A2 (en) Hydrophilic polymers with pendant functional groups and method thereof
EP3101064B1 (en) Diamine crosslinking agent, acidic polysaccharide crosslinked body, and medical material
CA3222952A1 (en) Dendrimer conjugates and methods of use thereof
JP5189243B2 (en) Drug complex and carrier for drug delivery

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 2018507416

Country of ref document: JP

NENP Non-entry into the national phase

Ref country code: DE

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17770374

Country of ref document: EP

Kind code of ref document: A1

122 Ep: pct application non-entry in european phase

Ref document number: 17770374

Country of ref document: EP

Kind code of ref document: A1