WO2012176823A1 - Anti-cancer agent composition - Google Patents

Anti-cancer agent composition Download PDF

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WO2012176823A1
WO2012176823A1 PCT/JP2012/065787 JP2012065787W WO2012176823A1 WO 2012176823 A1 WO2012176823 A1 WO 2012176823A1 JP 2012065787 W JP2012065787 W JP 2012065787W WO 2012176823 A1 WO2012176823 A1 WO 2012176823A1
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group
cyclodextrin
transition metal
substituent
agent composition
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PCT/JP2012/065787
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French (fr)
Japanese (ja)
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一博 中對
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日本化学工業株式会社
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Priority to CN201280024893.2A priority Critical patent/CN103547269B/en
Publication of WO2012176823A1 publication Critical patent/WO2012176823A1/en

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    • 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
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A61K47/40—Cyclodextrins; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/66—Phosphorus compounds
    • A61K31/675—Phosphorus compounds having nitrogen as a ring hetero atom, e.g. pyridoxal phosphate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C07F9/50—Organo-phosphines
    • C07F9/5004—Acyclic saturated phosphines
    • 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
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/645—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having two nitrogen atoms as the only ring hetero atoms
    • C07F9/6509—Six-membered rings
    • C07F9/650952—Six-membered rings having the nitrogen atoms in the positions 1 and 4
    • C07F9/650994—Six-membered rings having the nitrogen atoms in the positions 1 and 4 condensed with carbocyclic rings or carbocyclic ring systems

Definitions

  • the present invention relates to an anticancer agent composition containing a phosphine transition metal complex.
  • the present applicant has previously proposed an anticancer agent containing a phosphine transition metal complex represented by the following formula (1 ′) (see Patent Document 1).
  • R 1 , R 2 , R 3 and R 4 are alkyl groups and the like.
  • M is gold, copper or silver.
  • This anticancer agent has a high anticancer activity as compared with platinum preparations such as cisplatin and Taxol (registered trademark), which are conventionally used anticancer agents.
  • the phosphine transition metal complex Since the phosphine transition metal complex has low solubility in water, when the drug solution is administered to a living body, the precipitation of the phosphine transition metal complex in the body and the anticancer effect may be impaired. In addition, there is a problem that the form of administration to a living body becomes quite limited.
  • the subject of the present invention is to improve the above-mentioned anticancer agent proposed by the present applicant.
  • the present invention provides an anticancer agent composition
  • R 1 and R 2 represent a linear or branched alkyl group, a cycloalkyl group, a cycloalkyl group having a substituent, an adamantyl group, a phenyl group, or a phenyl group having a substituent;
  • the number may be the same or different
  • R 3 and R 4 each represents a hydrogen atom or a linear or branched alkyl group and has 1 carbon atom.
  • R 3 and R 4 may be bonded to each other to form a saturated or unsaturated ring, and the saturated or unsaturated group (The ring in (1) may have a substituent.
  • M represents a transition metal atom selected from the group consisting of gold, copper and silver, and X ⁇ represents an anion.
  • an anticancer agent composition having excellent solubility and high anticancer activity while having low toxicity is provided.
  • the anticancer composition of the present invention contains a phosphine transition metal complex represented by the above formula (1) and a cyclodextrin compound.
  • R 1 and R 2 are a linear or branched alkyl group, a cycloalkyl group, a cycloalkyl group having a substituent, an adamantyl group, a phenyl group, or a phenyl group having a substituent. Indicates. R 1 and R 2 have 1 to 10 carbon atoms. R 1 and R 2 may be the same group or different groups.
  • Examples of the alkyl group related to R 1 and R 2 include methyl group, ethyl group, isopropyl group, n-propyl group, isobutyl group, n-butyl group, sec-butyl group, tert-butyl group, isoheptyl group, n -Heptyl group, isohexyl group, n-hexyl group.
  • examples of the cycloalkyl group according to R 1 and R 2 include a cyclopentyl group and a cyclohexyl group.
  • R 1 and R 2 are a cycloalkyl group having a substituent or a phenyl group having a substituent
  • the substituents relating to the cycloalkyl group having a substituent or the phenyl group having a substituent include an alkyl group, nitro Group, amino group, hydroxyl group, fluoro group, chloro group, bromo group and iodo group.
  • R 1 is a t-butyl group and R 2 is a methyl group from the viewpoint of high anticancer activity.
  • R 3 and R 4 in the general formula (1) represent a hydrogen atom or a linear or branched alkyl group.
  • R 3 and R 4 have 1 to 6 carbon atoms.
  • R 3 and R 4 may be the same group or different groups.
  • R 3 and R 4 may be bonded to each other to form a saturated or unsaturated ring.
  • the unsaturated ring may have a substituent.
  • Examples of the alkyl group related to R 3 and R 4 include an ethyl group, an isopropyl group, an n-propyl group, an isobutyl group, an n-butyl group, a sec-butyl group, a tert-butyl group, an isoheptyl group, and an n-heptyl group. , Isohexyl group, n-hexyl group, cyclopentyl group, cyclohexyl group and the like.
  • the ring formed by combining R 3 and R 4 with each other is a saturated or unsaturated five-membered ring.
  • a 6-membered ring is mentioned, For example, a phenyl group, a cyclohexyl group, a cyclopentyl group etc. are mentioned.
  • the ring R 3 and R 4 are bonded to formed together may have a monovalent substituent, the monovalent with the rings R 3 and R 4 are bonded are formed by mutually substituted Examples of the group include linear or branched alkyl groups having 1 to 5 carbon atoms, nitro group, amino group, hydroxyl group, fluoro group, chloro group, bromo group, and iodo group.
  • M represents a transition metal atom selected from the group consisting of gold, copper and silver. And it is preferable that M is a gold atom at the point that anticancer activity becomes high.
  • X ⁇ represents an anion, and examples thereof include chlorine ion, bromine ion, iodine ion, boron tetrafluoride ion, hexafluorophosphate ion, and perchlorate ion.
  • X ⁇ is preferably a chlorine ion, a bromine ion or an iodine ion from the viewpoint of high anticancer activity.
  • R 3 and R 4 are bonded to each other to form a benzene ring from the viewpoint of high anticancer activity.
  • the phosphine transition metal complex when R 3 and R 4 are bonded to each other to form a benzene ring is a phosphine transition metal complex represented by the following general formula (2).
  • R 1, R 2, M and X - is, R 1 in the general formula (A), R 2, M and X - as synonymous.
  • R 5 represents a monovalent substituent.
  • substituents according to R 5 include linear or branched alkyl groups having 1 to 5 carbon atoms, nitro groups, amino groups, hydroxyl groups, fluoro groups, chloro groups, bromo groups, iodo groups.
  • Groups. N represents an integer of 0 to 4.
  • the phosphine transition metal complex represented by the general formula (1) is a group in which R 1 and R 2 are different from each other, the following general formula (3): (Wherein R 1 , R 2 , R 3 , R 4 , M and X ⁇ are the same as described above, * represents an asymmetric phosphorus atom), and a phosphine having an asymmetric center on the phosphorus atom It becomes a transition metal complex.
  • the phosphine transition metal complex represented by the general formula (3) has four asymmetric phosphorus atoms, there are many isomers.
  • the types of these isomers are as follows.
  • the phosphine transition metal complex may be a mixture containing isomers. Specifically, these isomers are composed of a single enantiomer, such as (R, R) (R, R) or (S, S) (S, S), on the phosphorus atom. Or may be composed of a racemic ligand, such as (R, R) (S, S), or as (R, S) (S, R).
  • each may be composed of a meso form, or may be composed of one enantiomer and its meso form, such as (R, R) (S, R).
  • This phosphine transition metal complex can be obtained by reacting a 2,3-bisphosphinopyrazine derivative represented by the formula (4) with a salt of gold, copper or silver.
  • the 2,3-bisphosphinopyrazine derivative represented by the formula (4) includes, for example, as shown in the reaction formula (9), 2,3-dichloropyrazine (6) and phosphine-borane (7). Reaction is carried out to obtain bis (phosphine-borane) pyrazine (8), and then the resulting bis (phosphine-borane) pyrazine (8) is subjected to a deboraneation reaction.
  • reaction formula (9) the reaction between 2,3-dichloropyrazine (6) and phosphine-borane (7) is carried out in the presence of a base such as n-butyllithium, sec-butyllithium, tetrahydrofuran, N , N-dimethylformamide and the like in an inert solvent at ⁇ 78 to 30 ° C. for 1 to 24 hours.
  • a base such as n-butyllithium, sec-butyllithium, tetrahydrofuran, N , N-dimethylformamide and the like in an inert solvent at ⁇ 78 to 30 ° C. for 1 to 24 hours.
  • 2,3-dichloropyrazine (6) and phosphine-borane (7) are produced by known methods.
  • 2,3-dichloropyrazine (6) is commercially available.
  • Phosphine-borane (7) is disclosed in, for example, JP-A No. 2003-3000988, JP-A No. 2001-253889, JP-A No. 2007-70310, J. Org. Chem. 2000, vol. 65, P4185-4188, etc. Manufactured using the methods described.
  • the deboraneation reaction of bis (phosphine-borane) pyrazine (8) is carried out by adding N, N, N ′, N ′,-tetramethylethylenediamine (TMEDA) to a reaction system containing bis (phosphine-borane) pyrazine (8). And the like, and a reaction is carried out at 0 to 100 ° C. for 10 minutes to 3 hours.
  • TEDA trimethylethylenediamine
  • this 2,3-bisphosphinopyrazine derivative represented by the formula (4), two phosphorus atoms become chiral centers because R 1 and R 2 are different groups.
  • this 2,3-bisphosphinopyrazine derivative has three types of isomers, (R, R), (S, S) and (R, S), which have different steric configurations.
  • the (R, S) isomer is a meso isomer
  • an equimolar mixture of (R, R) isomer and (S, S) isomer is a racemate.
  • Gold, copper or silver salts that react with the 2,3-bisphosphinopyrazine derivative represented by the formula (4) include, for example, halides, nitrates, perchlorates, and tetrafluoroboric acids of these metals. Salts, hexafluorophosphate and the like. Moreover, the valence of these metals is monovalent. In addition, these metal salts may be two or more salts in which either one or both of the metal and the anion are different.
  • Preferred gold transition metal salts include, for example, gold chloride (I) acid, gold chloride (I), or tetrabutylammonium chloride / gold chloride (I) (“5th edition, Experimental Chemistry Course 21”, editor) The Chemical Society of Japan, publisher Maruzen, publication date March 30, 2004, p. 366-380, see Aust. J. Chemm., 1997, 50, pages 775-778).
  • Preferred transition metal salts of copper include, for example, copper chloride (I), copper bromide (I), copper iodide (I), etc. (“Fifth edition Experimental Chemistry Course 21", Editor, The Chemical Society of Japan, published) Tokoro Maruzen, issue date March 30, 2004, p349-361).
  • Preferred silver transition metal salts include, for example, silver chloride (I), silver bromide (I), silver iodide (I), etc. ("5th edition Experimental Chemistry Course 21", Editor, The Chemical Society of Japan). Issue place Maruzen, issue date March 30, 2004, pages 361-366).
  • the transition metal salt according to the method for producing a phosphine transition metal complex of the present invention may be an anhydride or a hydrate.
  • the molar ratio of the 2,3-bisphosphinopyrazine derivative represented by the formula (4) to the salt of gold, copper or silver is preferably 1 to 5 times mol, more preferably 1.8 to 2.2 times mol.
  • the reaction can be carried out in a solvent such as acetone, acetonitrile, methanol, ethanol, tetrahydrofuran, dichloromethane, chloroform and the like.
  • the reaction temperature is preferably ⁇ 20 to 60 ° C., more preferably 0 to 25 ° C., and the reaction time is preferably 0.5 to 48 hours, more preferably 1 to 3 hours.
  • a phosphine transition metal complex represented by the formula (1) is obtained. After completion of the reaction, conventional purification can be performed as necessary.
  • the anion in the phosphine transition metal complex represented by the formula (1) thus obtained may be converted into another desired anion.
  • a phosphine transition metal complex in which X ⁇ in formula (1) is a halide ion is synthesized according to the production method described above, and then the phosphine transition metal complex and an inorganic acid having a desired anion,
  • a phosphine transition metal complex in which X ⁇ is a desired anion can be obtained by reacting an organic acid or an alkali metal salt thereof in a suitable solvent. Details of such a method are described in, for example, JP-A-10-147590, JP-A-10-114782, and JP-A-61-15944.
  • the phosphine transition metal complex is included in a cyclodextrin compound to form a complex (hereinafter referred to as “cyclodextrin inclusion body”).
  • cyclodextrin inclusion body The inclusion of the cyclodextrin compound is considered to involve the molecular diameter of the guest molecule to be included, the Van der Waals force with the guest molecule, and the hydrogen bond with the hydroxyl group derived from the cyclodextrin compound. It is difficult to predict the effects of cyclodextrins on specific compounds.
  • the cyclodextrin inclusion body can be prepared, for example, by preparing an aqueous solution of a cyclodextrin compound, adding the phosphine transition metal complex thereto, and stirring.
  • the compounding amount of the phosphine transition metal complex with respect to the cyclodextrin compound is 10 to 75 parts by weight, preferably 20 to 50 parts by weight with respect to 100 parts by weight of the cyclodextrin compound. This is preferable from the viewpoint of suppressing undissolved residue that does not participate in the inclusion of the complex.
  • the cyclodextrin clathrate thus obtained is used as an anticancer agent composition.
  • the anticancer agent composition of the present invention can be administered to humans or animals in various forms.
  • the administration form may be oral administration, or parenteral administration such as intravenous, intramuscular, subcutaneous or intradermal injection, rectal administration, transmucosal administration and the like.
  • Examples of the dosage form suitable for oral administration include tablets, pills, granules, powders, capsules, solutions, suspensions, emulsions, syrups and the like.
  • pharmaceutical compositions suitable for parenteral administration include injections, drops, nasal drops, sprays, inhalants, suppositories, ointments, creams, powdered coatings, liquid coatings, patches, etc. And the like.
  • examples of the dosage form of the anticancer agent of the present invention include an embedding pellet and a sustained-release preparation using a known technique.
  • preferred administration forms, preparation forms, and the like are appropriately selected by a doctor according to the age, sex, constitution, symptoms, treatment timing, etc. of the patient.
  • the anticancer composition of the present invention is a solid preparation such as a tablet, pill, powder, powder, granule, etc.
  • these solid preparations are suitable for the cyclodextrin inclusion body of the present invention according to a conventional method.
  • Additives such as lactose, sucrose, D-mannitol, corn starch, synthetic or natural gum, excipients such as crystalline cellulose, binders such as starch, hydroxypropylcellulose, hydroxypropylmethylcellulose, gum arabic, gelatin, polyvinylpyrrolidone , Carboxymethylcellulose calcium, sodium carboxymethylcellulose, starch, corn starch, sodium alginate and other disintegrants, lubricants such as talc, magnesium stearate, sodium stearate, calcium carbonate, sodium carbonate, calcium phosphate, sodium phosphate They are prepared by mixing as appropriate with fillers or diluents such as the like. Tablets and the like may be subjected to sugar coating, gelatin,
  • Liquid preparations include the cyclodextrin inclusion body of the present invention, purified water, a suitable buffer solution such as phosphate buffer, physiological saline solution such as physiological saline, Ringer's solution, lock solution, cocoa butter, sesame oil, Dissolved in vegetable oils such as olive oil, mineral oil, higher alcohols, higher fatty acids, ethanol and other organic solvents, etc., emulsifiers such as cholesterol, suspending agents such as gum arabic, dispersion aids, wetting agents, polyoxy if necessary Surfactants such as ethylene hydrogenated castor oil and polyethylene glycol, solubilizers such as sodium phosphate, stabilizers such as sugar, sugar alcohol and albumin, preservatives such as paraben, sodium chloride, Sterilized by appropriate addition of isotonic agents
  • the anticancer agent composition of the present invention is a semisolid preparation such as a lotion, cream or ointment
  • these semisolid preparations contain the cyclodextrin inclusion body of the present invention as a fat, fatty oil, lanolin or petrolatum.
  • the content of the cyclodextrin inclusion body of the present invention in the anticancer agent composition varies depending on the administration form, severity, target dose, etc.
  • the ratio of the total amount of the cyclodextrin inclusion body of the present invention to the total mass is preferably 0.001 to 90% by mass, more preferably 0.1 to 80% by mass.
  • the dose of the anticancer agent composition of the present invention is appropriately determined by a doctor according to conditions such as the age, sex, weight, symptom, and route of administration of a patient, but is generally an adult.
  • the amount of the active ingredient per day is in the range of about 1 ⁇ g / kg to 1,000 mg / kg, preferably in the range of about 10 ⁇ g / kg to 10 mg / kg as the phosphine transition metal complex.
  • the anticancer agent can be administered once a day, or can be administered in several divided doses (for example, about 2 to 4 times).
  • the type of cancer to which the anticancer composition of the present invention is applied is not particularly limited, and examples thereof include malignant melanoma, malignant lymphoma, digestive organ cancer, lung cancer, esophageal cancer, stomach cancer, colon cancer, Rectal cancer, colon cancer, ureteral tumor, gallbladder cancer, bile duct cancer, biliary tract cancer, breast cancer, liver cancer, pancreatic cancer, testicular cancer, maxillary cancer, tongue cancer, lip cancer, oral cancer, pharyngeal cancer, laryngeal cancer, ovarian cancer , Uterine cancer, prostate cancer, thyroid cancer, brain tumor, Kaposi's sarcoma, hemangioma, leukemia, polycythemia vera, neuroblastoma, retinoblastoma, myeloma, cystoma, sarcoma, osteosarcoma, muscle tumor, skin cancer, Examples include basal cell cancer, skin appendage cancer, skin metastatic cancer, cutaneous
  • the anticancer composition of the present invention can be combined with known chemotherapy, surgical treatment, radiation therapy, hyperthermia, immunotherapy and the like.
  • an aqueous solution prepared by dissolving potassium hydroxide (13.5 g, 240 mmol), potassium persulfate (19.4 g, 72.0 mmol) and ruthenium trichloride trihydrate (624 mg, 2.4 mmol) in 150 mL of water was prepared. Then, the acetone solution was gradually added at 0 ° C. while vigorously stirring the aqueous solution. After 2 hours, the reaction mixture was neutralized with 3M hydrochloric acid and extracted three times with ether. The organic phase was washed with saturated brine and dried over sodium sulfate.
  • Examples 1 to 8 ⁇ The amounts and types of cyclodextrin compounds shown in Table 1 were added to 4 ml of water. Next, 10 mg of the phosphine gold complex sample prepared above was added, mixed by stirring for 16 hours at 25 ° C., and various cyclodextrin inclusion bodies were obtained. Next, after standing for 2 hours, 1 mL of the supernatant was collected and centrifuged. An additional 0.7 ml of this supernatant was collected and centrifuged twice, and after centrifugation, 0.100 ml of the supernatant was collected and placed in a 5 ml volumetric flask and made up to 5 ml with methanol.
  • Example 9 (Preparation of ⁇ -cyclodextrin clathrate sample) 5 ml of a 2 wt% aqueous solution of ⁇ -cyclodextrin was prepared, 40 mg of a phosphine gold complex sample was added, and the mixture was stirred and mixed at 25 ° C. for 16 hours to prepare a ⁇ -cyclodextrin clathrate aqueous solution.
  • the cells were washed with PBS, and after counting the number of cells, a 1 ⁇ 10 6 cell / ml suspension was prepared using the same medium. The suspension was added to a sterile 96-well microplate to a density of 50000 cells / well. Next, ⁇ -cyclodextrin clathrate aqueous solution prepared above, cisplatin solution completely dissolved in dimethyl sulfoxide (Comparative Example 1), or phosphine gold complex sample completely dissolved in water without adding ⁇ -cyclodextrin (Reference Example 1) was added, followed by culturing in an incubator for 24 hours. Thereafter, the number of viable cells was evaluated by a modified method of Mosmann (T. Mosmann, J. Immunol.
  • ⁇ -cyclodextrin inclusion complex has higher anticancer activity than cisplatin, and even if phosphine gold complex is used as ⁇ -cyclodextrin inclusion complex, It can be seen that the high anticancer activity is maintained as it is.
  • an anticancer agent composition having excellent solubility and high anticancer activity while having low toxicity is provided.

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Abstract

Provided is an anti-cancer agent composition exhibiting excellent solubility and exerting high anti-cancer activity while exhibiting low toxicity. An anti-cancer composition characterized by containing a cyclodextrin compound and a phosphine transition metal complex represented by general formula (1). (In the formula, R1 and R2 represent a linear or branched alkyl group, a cycloalkyl group, a cycloalkyl group having a substituent, an adamantyl group, a phenyl group, or a phenyl group having a substituent, have a carbon number between 1 and 10, and may represent the same group or different groups. R3 and R4 represent a hydrogen atom or a linear or branched alkyl group, have a carbon number between 1 and 6, and may represent the same group or different groups. R3 and R4 may form a saturated or unsaturated ring by bonding to one another and said saturated or unsaturated ring may have a substituent. M represents a transition metal atom selected from among gold, copper, and silver. X- represents an anion.)

Description

抗がん剤組成物Anticancer composition
　　本発明は、ホスフィン遷移金属錯体を含有する抗がん剤組成物に関する。 The present invention relates to an anticancer agent composition containing a phosphine transition metal complex.
　本出願人は先に、以下の式（１’）で表されるホスフィン遷移金属錯体を含有する抗がん剤を提案した（特許文献１参照）。式中、Ｒ1、Ｒ2、Ｒ3、Ｒ4はアルキル基等である。
　またＭは、金、銅又は銀である。この抗がん剤は、従来用いられてきた抗がん剤であるシスプラチン等の白金製剤やタキソール（登録商標）に比較して、高い抗がん活性を有するものである。 The present applicant has previously proposed an anticancer agent containing a phosphine transition metal complex represented by the following formula (1 ′) (see Patent Document 1). In the formula, R 1 , R 2 , R 3 and R 4 are alkyl groups and the like.
M is gold, copper or silver. This anticancer agent has a high anticancer activity as compared with platinum preparations such as cisplatin and Taxol (registered trademark), which are conventionally used anticancer agents.
Figure JPOXMLDOC01-appb-C000003
Figure JPOXMLDOC01-appb-C000003
特開２００７－３２０９０９号公報JP 2007-320909 A
　前記ホスフィン遷移金属錯体は水に対して溶解性が低いため、薬剤溶液を生体に投与した場合に、体内でホスフィン遷移金属錯体の析出や、また抗がん効果が損なわれる可能がある。また、生体への投与形態もかなり限られたものになると言う問題もある。 Since the phosphine transition metal complex has low solubility in water, when the drug solution is administered to a living body, the precipitation of the phosphine transition metal complex in the body and the anticancer effect may be impaired. In addition, there is a problem that the form of administration to a living body becomes quite limited.
　上述の現況に鑑み、本発明の課題は、本出願人が提案した前述の抗がん剤の改良にある。 In view of the above-mentioned present situation, the subject of the present invention is to improve the above-mentioned anticancer agent proposed by the present applicant.
　本発明は、下記一般式（１）で表されるホスフィン遷移金属錯体と、シクロデキストリン化合物とを含有することを特徴とする抗がん剤組成物を提供するものである。
Figure JPOXMLDOC01-appb-C000004
 （式中、Ｒ１及びＲ２は、直鎖状若しくは分岐鎖状のアルキル基、シクロアルキル基、置換基を有するシクロアルキル基、アダマンチル基、フェニル基又は置換基を有するフェニル基を示し、炭素数が１～１０であり、同一の基であっても異なる基であってもよい。Ｒ３及びＲ４は、水素原子又は直鎖状若しくは分岐鎖状のアルキル基を示し、炭素数が１～６であり、同一の基であっても異なる基であってもよい。Ｒ３及びＲ４は、互いに結合して飽和又は不飽和の環を形成していてもよく、該飽和又は不飽和の環は、置換基を有していてもよい。Ｍは、金、銅及び銀の群から選ばれる遷移金属原子を示す。Ｘ－は、アニオンを示す。） The present invention provides an anticancer agent composition comprising a phosphine transition metal complex represented by the following general formula (1) and a cyclodextrin compound.
Figure JPOXMLDOC01-appb-C000004
 (Wherein R 1 and R 2 represent a linear or branched alkyl group, a cycloalkyl group, a cycloalkyl group having a substituent, an adamantyl group, a phenyl group, or a phenyl group having a substituent; The number may be the same or different, and R 3 and R 4 each represents a hydrogen atom or a linear or branched alkyl group and has 1 carbon atom. And may be the same group or different groups, and R 3 and R 4 may be bonded to each other to form a saturated or unsaturated ring, and the saturated or unsaturated group (The ring in (1) may have a substituent. M represents a transition metal atom selected from the group consisting of gold, copper and silver, and X − represents an anion.)
　本発明の抗がん剤組成物によれば、溶解性に優れ、高い抗がん活性を有しつつ、毒性の低い抗がん剤組成物が提供される。 According to the anticancer agent composition of the present invention, an anticancer agent composition having excellent solubility and high anticancer activity while having low toxicity is provided.
　本発明の抗がん剤組成物は、前記の式（１）で表されるホスフィン遷移金属錯体とシクロデキストリン化合物と、を含有している。 The anticancer composition of the present invention contains a phosphine transition metal complex represented by the above formula (1) and a cyclodextrin compound.
　前記一般式（１）中、Ｒ１及びＲ２は、直鎖状若しくは分岐鎖状のアルキル基、シクロアルキル基、置換基を有するシクロアルキル基、アダマンチル基、フェニル基又は置換基を有するフェニル基を示す。また、Ｒ１及びＲ２は、炭素数が１～１０である。また、Ｒ１及びＲ２は、同一の基であっても異なる基であってもよい。 In the general formula (1), R 1 and R 2 are a linear or branched alkyl group, a cycloalkyl group, a cycloalkyl group having a substituent, an adamantyl group, a phenyl group, or a phenyl group having a substituent. Indicates. R 1 and R 2 have 1 to 10 carbon atoms. R 1 and R 2 may be the same group or different groups.
　Ｒ１及びＲ２に係るアルキル基としては、例えば、メチル基、エチル基、イソプロピル基、ｎ－プロピル基、イソブチル基、ｎ－ブチル基、ｓｅｃ－ブチル基、ｔｅｒｔ－ブチル基、イソヘプチル基、ｎ－ヘプチル基、イソヘキシル基、ｎ－ヘキシル基が挙げられる。また、Ｒ１及びＲ２に係るシクロアルキル基としては、シクロペンチル基、シクロヘキシル基が挙げられる。また、Ｒ１及びＲ２が置換基を有するシクロアルキル基又は置換基を有するフェニル基の場合、置換基を有するシクロアルキル基又は置換基を有するフェニル基に係る置換基としては、アルキル基、ニトロ基、アミノ基、ヒドロキシル基、フルオロ基、クロロ基、ブロモ基、ヨード基が挙げられる。また、Ｒ１がｔ－ブチル基であり、Ｒ２がメチル基であることが、抗がん活性が高くなる点で好ましい。 Examples of the alkyl group related to R 1 and R 2 include methyl group, ethyl group, isopropyl group, n-propyl group, isobutyl group, n-butyl group, sec-butyl group, tert-butyl group, isoheptyl group, n -Heptyl group, isohexyl group, n-hexyl group. In addition, examples of the cycloalkyl group according to R 1 and R 2 include a cyclopentyl group and a cyclohexyl group. In the case where R 1 and R 2 are a cycloalkyl group having a substituent or a phenyl group having a substituent, the substituents relating to the cycloalkyl group having a substituent or the phenyl group having a substituent include an alkyl group, nitro Group, amino group, hydroxyl group, fluoro group, chloro group, bromo group and iodo group. In addition, it is preferable that R 1 is a t-butyl group and R 2 is a methyl group from the viewpoint of high anticancer activity.
　前記一般式（１）中のＲ３及びＲ４は、水素原子又は直鎖状若しくは分岐鎖状のアルキル基を示す。また、Ｒ３及びＲ４は、炭素数が１～６である。また、Ｒ３及びＲ４は、同一の基であっても異なる基であってもよい。また、Ｒ３及びＲ４は、互いに結合して飽和又は不飽和の環を形成していてもよく、Ｒ３及びＲ４が互いに結合して飽和又は不飽和の環を形成する場合、飽和又は不飽和の環は、置換基を有していてもよい。 R 3 and R 4 in the general formula (1) represent a hydrogen atom or a linear or branched alkyl group. R 3 and R 4 have 1 to 6 carbon atoms. R 3 and R 4 may be the same group or different groups. R 3 and R 4 may be bonded to each other to form a saturated or unsaturated ring. When R 3 and R 4 are bonded to each other to form a saturated or unsaturated ring, The unsaturated ring may have a substituent.
　Ｒ３及びＲ４に係るアルキル基としては、例えば、エチル基、イソプロピル基、ｎ－プロピル基、イソブチル基、ｎ－ブチル基、ｓｅｃ－ブチル基、ｔｅｒｔ－ブチル基、イソヘプチル基、ｎ－ヘプチル基、イソヘキシル基、ｎ－ヘキシル基、シクロペンチル基、シクロヘキシル基等が挙げられる。 Examples of the alkyl group related to R 3 and R 4 include an ethyl group, an isopropyl group, an n-propyl group, an isobutyl group, an n-butyl group, a sec-butyl group, a tert-butyl group, an isoheptyl group, and an n-heptyl group. , Isohexyl group, n-hexyl group, cyclopentyl group, cyclohexyl group and the like.
　また、Ｒ３及びＲ４が互いに結合して飽和又は不飽和の環を形成している場合、Ｒ３及びＲ４が互いに結合して形成された環としては、飽和又は不飽和の五員環又は六員環が挙げられ、例えば、フェニル基、シクロヘキシル基、シクロペンチル基等が挙げられる。また、Ｒ３及びＲ４が互いに結合して形成された環は、一価の置換基を有してもよく、Ｒ３及びＲ４が互いに結合して形成された環が有する一価の置換基としては、例えば、直鎖状又は分岐鎖状であり且つ炭素数が１～５のアルキル基、ニトロ基、アミノ基、ヒドロキシル基、フルオロ基、クロロ基、ブロモ基、ヨード基が挙げられる。 When R 3 and R 4 are bonded to each other to form a saturated or unsaturated ring, the ring formed by combining R 3 and R 4 with each other is a saturated or unsaturated five-membered ring. Or a 6-membered ring is mentioned, For example, a phenyl group, a cyclohexyl group, a cyclopentyl group etc. are mentioned. The ring R 3 and R 4 are bonded to formed together may have a monovalent substituent, the monovalent with the rings R 3 and R 4 are bonded are formed by mutually substituted Examples of the group include linear or branched alkyl groups having 1 to 5 carbon atoms, nitro group, amino group, hydroxyl group, fluoro group, chloro group, bromo group, and iodo group.
　前記一般式（１）中、Ｍは、金、銅及び銀の群から選ばれる遷移金属原子を示す。そして、Ｍが金原子であることが抗がん活性が高くなる点で好ましい。 In the general formula (1), M represents a transition metal atom selected from the group consisting of gold, copper and silver. And it is preferable that M is a gold atom at the point that anticancer activity becomes high.
　前記一般式（１）中、Ｘ－は、アニオンを示し、例えば、塩素イオン、臭素イオン、ヨウ素イオン、四フッ化ホウ素イオン、六フッ化リン酸イオン、過塩素酸イオン等が挙げられる。これらのうち、Ｘ－が、塩素イオン、臭素イオン、ヨウ素イオンであることが、抗がん活性が高くなる点で好ましい。 In the general formula (1), X − represents an anion, and examples thereof include chlorine ion, bromine ion, iodine ion, boron tetrafluoride ion, hexafluorophosphate ion, and perchlorate ion. Of these, X − is preferably a chlorine ion, a bromine ion or an iodine ion from the viewpoint of high anticancer activity.
　前記一般式（１）で表されるホスフィン遷移金属錯体において、Ｒ３及びＲ４が互いに結合してベンゼン環を形成していることが、抗がん活性が高くなる点で好ましい。 In the phosphine transition metal complex represented by the general formula (1), it is preferable that R 3 and R 4 are bonded to each other to form a benzene ring from the viewpoint of high anticancer activity.
　Ｒ３及びＲ４が互いに結合してベンゼン環を形成している場合のホスフィン遷移金属錯体とは、下記一般式（２）で表されるホスフィン遷移金属錯体である。
Figure JPOXMLDOC01-appb-C000005
 The phosphine transition metal complex when R 3 and R 4 are bonded to each other to form a benzene ring is a phosphine transition metal complex represented by the following general formula (2).
Figure JPOXMLDOC01-appb-C000005
　前記一般式（２）中、Ｒ１、Ｒ２、Ｍ及びＸ－は、前記一般式（Ａ）中のＲ１、Ｒ２、Ｍ及びＸ－と同義である。 In the general formula (2), R 1, R 2, M and X - is, R 1 in the general formula (A), R 2, M and X - as synonymous.
　前記一般式（２）中、Ｒ５は、一価の置換基を示す。Ｒ５に係る置換基としては、例えば、直鎖状又は分岐鎖状であり且つ炭素数が１～５のアルキル基、ニトロ基、アミノ基、ヒドロキシル基、フルオロ基、クロロ基、ブロモ基、ヨード基が挙げられる。また、ｎは、０～４の整数を示す。 In the general formula (2), R 5 represents a monovalent substituent. Examples of the substituent according to R 5 include linear or branched alkyl groups having 1 to 5 carbon atoms, nitro groups, amino groups, hydroxyl groups, fluoro groups, chloro groups, bromo groups, iodo groups. Groups. N represents an integer of 0 to 4.
　本発明において、前記一般式（１）で表されるホスフィン遷移金属錯体は、式中のＲ１とＲ２が互いに異なる基の場合、下記一般式（３）：
Figure JPOXMLDOC01-appb-C000006
 （式中、Ｒ１、Ｒ２、Ｒ３、Ｒ４、Ｍ及びＸ－は前記と同じ。＊は不斉リン原子を示す。）で表される、リン原子上に不斉中心を有するホスフィン遷移金属錯体になる。 In the present invention, when the phosphine transition metal complex represented by the general formula (1) is a group in which R 1 and R 2 are different from each other, the following general formula (3):
Figure JPOXMLDOC01-appb-C000006
 (Wherein R 1 , R 2 , R 3 , R 4 , M and X − are the same as described above, * represents an asymmetric phosphorus atom), and a phosphine having an asymmetric center on the phosphorus atom It becomes a transition metal complex.
　前記一般式（３）で表されるホスフィン遷移金属錯体は、不斉なリン原子を４個有するため、数多くの異性体が存在するが、本発明においては、これらの異性体の種類については、特に制限されるものではない。また、ホスフィン遷移金属錯体は、異性体を含む混合物であってもよい。具体的には、これらの異性体は、リン原子上の立体が、（Ｒ，Ｒ）（Ｒ，Ｒ）や、（Ｓ，Ｓ）（Ｓ，Ｓ）のように、単一のエナンチオマーから構成されていてもよく、また、（Ｒ，Ｒ）（Ｓ，Ｓ）のように、配位子のラセミ体から構成されていてもよく、また、（Ｒ，Ｓ）（Ｓ，Ｒ）のように、お互いにメソ体から構成されていてもよく、また、（Ｒ，Ｒ）（Ｓ，Ｒ）のように、１つのエナンチオマーとそのメソ体から構成されていてもよい。 Since the phosphine transition metal complex represented by the general formula (3) has four asymmetric phosphorus atoms, there are many isomers. In the present invention, the types of these isomers are as follows. There is no particular limitation. The phosphine transition metal complex may be a mixture containing isomers. Specifically, these isomers are composed of a single enantiomer, such as (R, R) (R, R) or (S, S) (S, S), on the phosphorus atom. Or may be composed of a racemic ligand, such as (R, R) (S, S), or as (R, S) (S, R). In addition, each may be composed of a meso form, or may be composed of one enantiomer and its meso form, such as (R, R) (S, R).
　異性体を含む混合物の一例として、１）下記の式（１ａ）～式（１ｄ）で表される化合物を２種以上を組み合わせもの、２）下記の式（１ａ）～（１ｄ）で表される化合物を２種以上含有することに加えて、下記の式（２ａ）～式（２ｃ）で表される化合物の群から選択される少なくもとも１種以上の化合物を含有するもの、３）下記の式（１ａ）～（１ｄ）で表される化合物を２種以上と、下記の式（Ａ）で表される化合物を含有するもの、４）下記の式（１ａ）～（１ｄ）で表される化合物を２種以上、下記の式（２ａ）～式（２ｃ）で表される化合物の群から選択される少なくもとも１種以上及び下記の式（Ａ）で表される化合物を含有するもの、或いは５）下記の式（２ａ）～（２ｃ）で表される化合物の群から選択される少なくとも１種と、下記の式（Ａ）で表される化合物を含有するもの等が挙げられる。 As an example of a mixture containing isomers, 1) a combination of two or more compounds represented by the following formulas (1a) to (1d), and 2) represented by the following formulas (1a) to (1d) In addition to containing at least one compound selected from the group of compounds represented by the following formulas (2a) to (2c), 3) A compound containing two or more compounds represented by the following formulas (1a) to (1d) and a compound represented by the following formula (A): 4) In the following formulas (1a) to (1d) At least one compound selected from the group of compounds represented by the following formulas (2a) to (2c) and a compound represented by the following formula (A): Or 5) at least selected from the group of compounds represented by the following formulas (2a) to (2c) Even with one, and the like which contains a compound represented by formula (A) below.
Figure JPOXMLDOC01-appb-C000007
Figure JPOXMLDOC01-appb-C000007
Figure JPOXMLDOC01-appb-C000008
Figure JPOXMLDOC01-appb-C000008
Figure JPOXMLDOC01-appb-C000009
Figure JPOXMLDOC01-appb-C000009
　次に、本発明の抗がん剤組成物に含まれる前記一般式（１）で表されるホスフィン遷移金属錯体の好適な製造方法について説明する。このホスフィン遷移金属錯体は、式（４）で表される２，３－ビスホスフィノピラジン誘導体と、金、銅又は銀の塩とを反応させることで得られる。
Figure JPOXMLDOC01-appb-C000010
 Next, the suitable manufacturing method of the phosphine transition metal complex represented by the said General formula (1) contained in the anticancer agent composition of this invention is demonstrated. This phosphine transition metal complex can be obtained by reacting a 2,3-bisphosphinopyrazine derivative represented by the formula (4) with a salt of gold, copper or silver.
Figure JPOXMLDOC01-appb-C000010
　式（４）で表される２，３－ビスホスフィノピラジン誘導体は、例えば、反応式（９）に示すように、２，３－ジクロロピラジン（６）と、ホスフィン―ボラン（７）とを反応させ、ビス（ホスフィン－ボラン）ピラジン（８）を得、次いで、得られたビス（ホスフィン－ボラン）ピラジン（８）の脱ボラン化反応を行うことにより製造される。
Figure JPOXMLDOC01-appb-C000011
 The 2,3-bisphosphinopyrazine derivative represented by the formula (4) includes, for example, as shown in the reaction formula (9), 2,3-dichloropyrazine (6) and phosphine-borane (7). Reaction is carried out to obtain bis (phosphine-borane) pyrazine (8), and then the resulting bis (phosphine-borane) pyrazine (8) is subjected to a deboraneation reaction.
Figure JPOXMLDOC01-appb-C000011
　反応式（９）において、２，３－ジクロロピラジン（６）と、ホスフィン－ボラン（７）との反応は、例えば、ｎ－ブチルリチウム、ｓｅｃ－ブチルリチウム等の塩基の存在下、テトラヒドロフランやＮ，Ｎ－ジメチルホルムアミド等の不活性溶媒中、－７８～３０℃で、１～２４時間反応させることにより行なわれる。 In the reaction formula (9), the reaction between 2,3-dichloropyrazine (6) and phosphine-borane (7) is carried out in the presence of a base such as n-butyllithium, sec-butyllithium, tetrahydrofuran, N , N-dimethylformamide and the like in an inert solvent at −78 to 30 ° C. for 1 to 24 hours.
　２，３－ジクロロピラジン（６）及びホスフィン－ボラン（７）は、公知の方法により製造される。２，３－ジクロロピラジン（６）は、市販されている。ホスフィン－ボラン（７）は、例えば特開２００３－３００９８８号公報、特開２００１－２５３８８９号公報、特開２００７－７０３１０号公報及びJ.Org.Chem,2000,vol.65,P4185-4188等に記載されている方法を用いて製造される。 2,3-dichloropyrazine (6) and phosphine-borane (7) are produced by known methods. 2,3-dichloropyrazine (6) is commercially available. Phosphine-borane (7) is disclosed in, for example, JP-A No. 2003-3000988, JP-A No. 2001-253889, JP-A No. 2007-70310, J. Org. Chem. 2000, vol. 65, P4185-4188, etc. Manufactured using the methods described.
　ビス（ホスフィン－ボラン）ピラジン（８）の脱ボラン化反応は、ビス（ホスフィン－ボラン）ピラジン（８）を含有する反応系に、Ｎ，Ｎ，Ｎ’，Ｎ’，－テトラメチルエチレンジアミン（ＴＭＥＤＡ）等の脱ボラン化剤を添加し、０～１００℃で、１０分～３時間反応させることにより行なわれる。 The deboraneation reaction of bis (phosphine-borane) pyrazine (8) is carried out by adding N, N, N ′, N ′,-tetramethylethylenediamine (TMEDA) to a reaction system containing bis (phosphine-borane) pyrazine (8). And the like, and a reaction is carried out at 0 to 100 ° C. for 10 minutes to 3 hours.
　式（４）で表される２，３－ビスホスフィノピラジン誘導体においては、Ｒ１とＲ２が異なる基であることによって、２つのリン原子がキラル中心となる。その結果、この２，３－ビスホスフィノピラジン誘導体は、立体配置の異なる（Ｒ，Ｒ）体、（Ｓ，Ｓ）体及び（Ｒ，Ｓ）体の３種類の異性体が存在する。これらの３種類の異性体のうち、（Ｒ，Ｓ）体はメソ体であり、（Ｒ，Ｒ）体と（Ｓ，Ｓ）体の等モル混合物がラセミ体となる。こらの３種類の異性体を適切な量で用いることで、目的とする立体構造を有するホスフィン遷移金属錯体を得ることができる。 In the 2,3-bisphosphinopyrazine derivative represented by the formula (4), two phosphorus atoms become chiral centers because R 1 and R 2 are different groups. As a result, this 2,3-bisphosphinopyrazine derivative has three types of isomers, (R, R), (S, S) and (R, S), which have different steric configurations. Of these three types of isomers, the (R, S) isomer is a meso isomer, and an equimolar mixture of (R, R) isomer and (S, S) isomer is a racemate. By using these three types of isomers in appropriate amounts, a phosphine transition metal complex having the desired steric structure can be obtained.
　式（４）で表される２，３－ビスホスフィノピラジン誘導体と反応する金、銅又は銀の塩は、例えば、これらの金属のハロゲン化物、硝酸塩、過塩素酸塩、四フッ化ホウ素酸塩、六フッ化リン酸塩等である。また、これらの金属の価数は、一価である。また、これらの金属の塩は、金属又はアニオンのいずれか一方又は両方が異なる２種以上の塩であってもよい。 Gold, copper or silver salts that react with the 2,3-bisphosphinopyrazine derivative represented by the formula (4) include, for example, halides, nitrates, perchlorates, and tetrafluoroboric acids of these metals. Salts, hexafluorophosphate and the like. Moreover, the valence of these metals is monovalent. In addition, these metal salts may be two or more salts in which either one or both of the metal and the anion are different.
　好ましい金の遷移金属塩としては、例えば、塩化金（Ｉ）酸、塩化金（Ｉ）、あるいはテトラブチルアンモニウムクロリド・塩化金（Ｉ）等(「第５版　実験化学講座２１」、編者　社団法人日本化学会、発行所　丸善、発行日　平成１６年３月３０日、ｐ３６６～３８０、Aust.J.Chemm.,1997,50,775-778頁参照)が挙げられる。好ましい銅の遷移金属塩としては、例えば、塩化銅（Ｉ）、臭化銅（Ｉ）、ヨウ化銅（Ｉ）等（「第５版　実験化学講座２１」、編者　社団法人日本化学会、発行所　丸善、発行日　平成１６年３月３０日、ｐ３４９～３６１）が挙げられる。また、好ましい銀の遷移金属塩としては、例えば
、塩化銀（Ｉ）、臭化銀（Ｉ）、ヨウ化銀（Ｉ）等（「第５版　実験化学講座２１」、編者　社団法人日本化学会、発行所　丸善、発行日　平成１６年３月３０日、ｐ３６１～３６６）が挙げられる。なお、本発明のホスフィン遷移金属錯体の製造方法に係る遷移金属塩は、無水物であっても含水物であってもよい。 Preferred gold transition metal salts include, for example, gold chloride (I) acid, gold chloride (I), or tetrabutylammonium chloride / gold chloride (I) (“5th edition, Experimental Chemistry Course 21”, editor) The Chemical Society of Japan, publisher Maruzen, publication date March 30, 2004, p. 366-380, see Aust. J. Chemm., 1997, 50, pages 775-778). Preferred transition metal salts of copper include, for example, copper chloride (I), copper bromide (I), copper iodide (I), etc. ("Fifth edition Experimental Chemistry Course 21", Editor, The Chemical Society of Japan, published) Tokoro Maruzen, issue date March 30, 2004, p349-361). Preferred silver transition metal salts include, for example, silver chloride (I), silver bromide (I), silver iodide (I), etc. ("5th edition Experimental Chemistry Course 21", Editor, The Chemical Society of Japan). Issue place Maruzen, issue date March 30, 2004, pages 361-366). The transition metal salt according to the method for producing a phosphine transition metal complex of the present invention may be an anhydride or a hydrate.
　金、銅又は銀の塩に対する、式（４）で表わされる２，３－ビスホスフィノピラジン誘導体のモル比は、好ましくは１～５倍モル、更に好ましくは１．８～２．２倍モルとする。反応は、アセトン、アセトニトリル、メタノール、エタノール、テトラヒドロフラン、ジクロロメタン、クロロホルム等の溶媒中で行うことができる。反応温度は好ましくは－２０～６０℃、更に好ましくは０～２５℃であり、反応時間は好ましくは０．５～４８時間、更に好ましくは１～３時間である。この反応によって、式（１）で表されるホスフィン遷移金属錯体が得られる。反応終了後は、必要に応じて常法の精製を行うことができる。 The molar ratio of the 2,3-bisphosphinopyrazine derivative represented by the formula (4) to the salt of gold, copper or silver is preferably 1 to 5 times mol, more preferably 1.8 to 2.2 times mol. And The reaction can be carried out in a solvent such as acetone, acetonitrile, methanol, ethanol, tetrahydrofuran, dichloromethane, chloroform and the like. The reaction temperature is preferably −20 to 60 ° C., more preferably 0 to 25 ° C., and the reaction time is preferably 0.5 to 48 hours, more preferably 1 to 3 hours. By this reaction, a phosphine transition metal complex represented by the formula (1) is obtained. After completion of the reaction, conventional purification can be performed as necessary.
　このようにして得られた式（１）で表されるホスフィン遷移金属錯体におけるアニオンを、他の所望のアニオンに変換してもよい。例えば、先ず、上述の製造方法に従い、式（１）中のＸ-が、ハロゲン化物イオンであるホスフィン遷移金属錯体を合成し、次いで、このホスフィン遷移金属錯体と、所望のアニオンを有する無機酸、有機酸又はそれらのアルカリ金属塩とを、適切な溶媒中で反応させることにより、Ｘ-が、所望のアニオンであるホスフィン遷移金属錯体を得ることができる。このような方法の詳細は、例えば特開平１０－１４７５９０号公報、特開平１０－１１４７８２号公報及び特開昭６１－１０５９４号公報等に記載されている。 The anion in the phosphine transition metal complex represented by the formula (1) thus obtained may be converted into another desired anion. For example, first, a phosphine transition metal complex in which X − in formula (1) is a halide ion is synthesized according to the production method described above, and then the phosphine transition metal complex and an inorganic acid having a desired anion, A phosphine transition metal complex in which X − is a desired anion can be obtained by reacting an organic acid or an alkali metal salt thereof in a suitable solvent. Details of such a method are described in, for example, JP-A-10-147590, JP-A-10-114782, and JP-A-61-15944.
　前記ホスフィン遷移金属錯体は、シクロデキストリン化合物に包接され、複合体を形成する（以下、「シクロデキストリン包接体」という。）。シクロデキストリン化合物の包接は、包接するゲスト分子の分子径、ゲスト分子とのＶａｎ　ｄｅｒ　Ｗａａｌｓ力やシクロデキストリン化合物由来のヒドロキシル基との水素結合が関与しているものと考えられ、不溶性化合物ならすべてに適応できるものではなく、また、特定の化合物に対するシクロデキストリンの効果も予測することは困難である。 The phosphine transition metal complex is included in a cyclodextrin compound to form a complex (hereinafter referred to as “cyclodextrin inclusion body”). The inclusion of the cyclodextrin compound is considered to involve the molecular diameter of the guest molecule to be included, the Van der Waals force with the guest molecule, and the hydrogen bond with the hydroxyl group derived from the cyclodextrin compound. It is difficult to predict the effects of cyclodextrins on specific compounds.
　本発明で使用できるシクロデキストリン化合物としては、例えばα―シクロデキストリン、β－シクロデキストリン、γ―シクロデキストリン、メチル－β－シクロデキストリン、ジメチル－β－シクロデキストリン、トリメチル－β－シクロデキストリン、ペルメチル－β－シクロデキストリン、２－ヒドロキシプロピル－β―シクロデキストリン、２－ヒドロキシプロピル－α－シクロデキストリン、ジヒドロキシプロピル－β－シクロデキストリン、ヒドロキシエチル－β－シクロデキストリン、カルボキシメチル－β－シクロデキストリン、モノアセチル－β―シクロデキストリン、２－ヒドロキシプロピル－γ－シクロデキストリン、グルコシル－β－シクロデキストリン、マルトシル－α－シクロデキストリン、マルトシル－β－シクロデキストリン、パーシャリーメチル－β－シクロデキストリン、α－シクロデキストリンスルフェート、β－シクロデキストリンスルフェート、６－スルホヘキシル－β―シクロデキストリン、５－スルホペンチル－β―シクロデキストリン、４－スルホブチル－β―シクロデキストリン、３－スルホプロピル－β―シクロデキストリン、２－スルホエチル－β―シクロデキストリン等が挙げられ、このうちα―シクロデキストリン、β－シクロデキストリン、γ―シクロデキストリンが好ましく、特にβ－シクロデキストリンが、少ない添加量で溶解性をいっそう向上させることができる観点から好ましい。 Examples of cyclodextrin compounds that can be used in the present invention include α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, methyl-β-cyclodextrin, dimethyl-β-cyclodextrin, trimethyl-β-cyclodextrin, permethyl- β-cyclodextrin, 2-hydroxypropyl-β-cyclodextrin, 2-hydroxypropyl-α-cyclodextrin, dihydroxypropyl-β-cyclodextrin, hydroxyethyl-β-cyclodextrin, carboxymethyl-β-cyclodextrin, mono Acetyl-β-cyclodextrin, 2-hydroxypropyl-γ-cyclodextrin, glucosyl-β-cyclodextrin, maltosyl-α-cyclodextrin, maltosyl-β-cyclode Xistrin, partial methyl-β-cyclodextrin, α-cyclodextrin sulfate, β-cyclodextrin sulfate, 6-sulfohexyl-β-cyclodextrin, 5-sulfopentyl-β-cyclodextrin, 4-sulfobutyl-β -Cyclodextrin, 3-sulfopropyl-β-cyclodextrin, 2-sulfoethyl-β-cyclodextrin, etc., among which α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin are preferred, and β-cyclodextrin is particularly preferred. Dextrin is preferred from the viewpoint that the solubility can be further improved with a small addition amount.
　シクロデキストリン包接体は、例えばシクロデキストリン化合物の水溶液を調製し、これに前記ホスフィン遷移金属錯体を添加し攪拌することで調製することができる。シクロデキストリン化合物に対するホスフィン遷移金属錯体の配合量は、シクロデキストリン化合物１００重量部に対してホスフィン遷移金属錯体１０～７５重量部、好ましくは２０～５０重量部とすることがシクロデキストリン化合物及びホスフィン遷移金属錯体の包接に関与しない溶け残りを抑制する観点から好ましい。 The cyclodextrin inclusion body can be prepared, for example, by preparing an aqueous solution of a cyclodextrin compound, adding the phosphine transition metal complex thereto, and stirring. The compounding amount of the phosphine transition metal complex with respect to the cyclodextrin compound is 10 to 75 parts by weight, preferably 20 to 50 parts by weight with respect to 100 parts by weight of the cyclodextrin compound. This is preferable from the viewpoint of suppressing undissolved residue that does not participate in the inclusion of the complex.
　このようにして得られたシクロデキストリン包接体は抗がん剤組成物として用いられる。 The cyclodextrin clathrate thus obtained is used as an anticancer agent composition.
　本発明の抗がん剤組成物の使用においては、種々の形態でヒト又は動物に、本発明の抗がん剤組成物を投与することができる。投与形態としては、経口投与でもよいし、静脈内、筋肉内、皮下又は皮内等への注射、直腸内投与、経粘膜投与等の非経口投与でもよい。経口投与に適する製剤形態としては、例えば錠剤、丸剤、顆粒剤、散剤、カプセル剤、液剤、懸濁剤、乳剤、シロップ剤などを挙げることができる。非経口投与に適する医薬組成物としては、例えば、注射剤、点滴剤、点鼻剤、噴霧剤、吸入剤、坐剤、あるいは、軟膏、クリーム、粉状塗布剤、液状塗布剤、貼付剤等の経皮吸収剤等が挙げられる。更に、本発明の抗がん剤の製剤形態として、埋め込み用ペレットや公知の技術を用いた持続性製剤が挙げられる。 In the use of the anticancer agent composition of the present invention, the anticancer agent composition of the present invention can be administered to humans or animals in various forms. The administration form may be oral administration, or parenteral administration such as intravenous, intramuscular, subcutaneous or intradermal injection, rectal administration, transmucosal administration and the like. Examples of the dosage form suitable for oral administration include tablets, pills, granules, powders, capsules, solutions, suspensions, emulsions, syrups and the like. Examples of pharmaceutical compositions suitable for parenteral administration include injections, drops, nasal drops, sprays, inhalants, suppositories, ointments, creams, powdered coatings, liquid coatings, patches, etc. And the like. Furthermore, examples of the dosage form of the anticancer agent of the present invention include an embedding pellet and a sustained-release preparation using a known technique.
　上述したうち、好ましい投与形態や製剤形態等は、患者の年齢、性別、体質、症状、処置時期等に応じて、医師によって適宜選択される。 Among the above, preferred administration forms, preparation forms, and the like are appropriately selected by a doctor according to the age, sex, constitution, symptoms, treatment timing, etc. of the patient.
　本発明の抗がん剤組成物が、錠剤、丸剤、散剤、粉剤、顆粒剤等の固形製剤の場合、これらの固形製剤は、本発明のシクロデキストリン包接体を、常法に従って適当な添加剤、例えば乳糖、ショ糖、Ｄ－マンニトール、トウモロコシデンプン、合成若しくは天然ガム、結晶セルロース等の賦形剤、デンプン、ヒドロキシプロピルセルロース、ヒドロキシプロピルメチルセルロース、アラビアゴム、ゼラチン、ポリビニルピロリドン等の結合剤、カルボキシメチルセルロースカルシウム、カルボキシメチルセルロースナトリウム、デンプン、コーンスターチ、アルギン酸ナトリウム等の崩壊剤、タルク、ステアリン酸マグネシウム、ステアリン酸ナトリウム等の滑沢剤、炭酸カルシウム、炭酸ナトリウム、リン酸カルシウム、リン酸ナトリウム等の充填剤又は希釈剤等と適宜混合して製造される。錠剤等は、必要に応じて、ヒドロキシプロピルメチルセルロース、白糖、ポリエチレングリコール、酸化チタン等のコーティング剤を用いて、糖衣、ゼラチン、腸溶被覆、フイルムコーティング等が施されても良い。 When the anticancer composition of the present invention is a solid preparation such as a tablet, pill, powder, powder, granule, etc., these solid preparations are suitable for the cyclodextrin inclusion body of the present invention according to a conventional method. Additives such as lactose, sucrose, D-mannitol, corn starch, synthetic or natural gum, excipients such as crystalline cellulose, binders such as starch, hydroxypropylcellulose, hydroxypropylmethylcellulose, gum arabic, gelatin, polyvinylpyrrolidone , Carboxymethylcellulose calcium, sodium carboxymethylcellulose, starch, corn starch, sodium alginate and other disintegrants, lubricants such as talc, magnesium stearate, sodium stearate, calcium carbonate, sodium carbonate, calcium phosphate, sodium phosphate They are prepared by mixing as appropriate with fillers or diluents such as the like. Tablets and the like may be subjected to sugar coating, gelatin, enteric coating, film coating, and the like, if necessary, using a coating agent such as hydroxypropylmethylcellulose, sucrose, polyethylene glycol, and titanium oxide.
　本発明の抗がん剤組成物が、注射剤、点眼剤、点鼻剤、吸入剤、噴霧剤、ローション剤、シロップ剤、液剤、懸濁剤、乳剤等の液状製剤である場合、これらの液状製剤は、本発明のシクロデキストリン包接体を、精製水、リン酸緩衝液等の適当な緩衝液、生理的食塩水、リンゲル溶液、ロック溶液等の生理的塩類溶液、カカオバター、ゴマ油、オリーブ油等の植物油、鉱油、高級アルコール、高級脂肪酸、エタノール等の有機溶媒等に溶解して、必要に応じてコレステロール等の乳化剤、アラビアゴム等の懸濁剤、分散助剤、浸潤剤、ポリオキシエチレン硬化ヒマシ油系、ポリエチレングリコール系等の界面活性剤、リン酸ナトリウム等の溶解補助剤、糖、糖アルコール、アルブミン等の安定化剤、パラベン等の保存剤、塩化ナトリウム、ブドウ糖、グリセリン等の等張化剤、緩衝剤、無痛化剤、吸着防止剤、保湿剤、酸化防止剤、着色剤、甘味料、フレーバー、芳香物質等を適宜添加することにより、滅菌された水溶液、非水溶液、懸濁液、リポソーム又はエマルジョン等として調整される。この際、注射剤は、生理学的なｐＨを有することが好ましく、６～８の範囲内のｐＨを有することが特に好ましい。 When the anticancer composition of the present invention is a liquid preparation such as an injection, eye drops, nasal drop, inhalant, spray, lotion, syrup, liquid, suspension, emulsion, etc. Liquid preparations include the cyclodextrin inclusion body of the present invention, purified water, a suitable buffer solution such as phosphate buffer, physiological saline solution such as physiological saline, Ringer's solution, lock solution, cocoa butter, sesame oil, Dissolved in vegetable oils such as olive oil, mineral oil, higher alcohols, higher fatty acids, ethanol and other organic solvents, etc., emulsifiers such as cholesterol, suspending agents such as gum arabic, dispersion aids, wetting agents, polyoxy if necessary Surfactants such as ethylene hydrogenated castor oil and polyethylene glycol, solubilizers such as sodium phosphate, stabilizers such as sugar, sugar alcohol and albumin, preservatives such as paraben, sodium chloride, Sterilized by appropriate addition of isotonic agents such as dough sugar and glycerin, buffers, soothing agents, adsorption inhibitors, moisturizers, antioxidants, colorants, sweeteners, flavors, fragrances, etc. It is prepared as an aqueous solution, non-aqueous solution, suspension, liposome or emulsion. At this time, the injection preferably has a physiological pH, and particularly preferably has a pH within the range of 6-8.
　本発明の抗がん剤組成物が、ローション剤、クリーム剤、軟膏等の半固形製剤の場合、これらの半固形製剤は、本発明のシクロデキストリン包接体を脂肪、脂肪油、ラノリン、ワセリン、パラフィン、蝋、硬膏剤、樹脂、プラスチック、グリコール類、高級アルコール、グリセリン、水、乳化剤、懸濁化剤等と適宜混和することにより製造される。 When the anticancer agent composition of the present invention is a semisolid preparation such as a lotion, cream or ointment, these semisolid preparations contain the cyclodextrin inclusion body of the present invention as a fat, fatty oil, lanolin or petrolatum. , Paraffin, wax, plaster, resin, plastic, glycols, higher alcohol, glycerin, water, emulsifier, suspending agent and the like.
　抗がん剤組成物中の本発明のシクロデキストリン包接体の含有量は、投与形態、重篤度や目的とする投与量などによって様々であるが、一般的には、抗がん剤の全質量に対して、本発明のシクロデキストリン包接体の全量の割合が、好ましくは０．００１～９０質量％、更に好ましくは０．１～８０質量％である。 The content of the cyclodextrin inclusion body of the present invention in the anticancer agent composition varies depending on the administration form, severity, target dose, etc. The ratio of the total amount of the cyclodextrin inclusion body of the present invention to the total mass is preferably 0.001 to 90% by mass, more preferably 0.1 to 80% by mass.
　本発明の抗がん剤組成物の投与量は、例えば患者の年齢、性別、体重、症状及び投与経路などの条件に応じて適宜医師により決定されるものであるが、一般的には、成人一日あたりの有効成分の量として、ホスフィン遷移金属錯体として１μｇ／ｋｇから１，０００ｍｇ／ｋｇ程度の範囲であり、好ましくは１０μｇ／ｋｇから１０ｍｇ／ｋｇ程度の範囲である。この投与量の範囲内において、抗がん剤を一日一回で投与することができ、あるいは数回（例えば、２～４回程度）に分けて投与することができる。 The dose of the anticancer agent composition of the present invention is appropriately determined by a doctor according to conditions such as the age, sex, weight, symptom, and route of administration of a patient, but is generally an adult. The amount of the active ingredient per day is in the range of about 1 μg / kg to 1,000 mg / kg, preferably in the range of about 10 μg / kg to 10 mg / kg as the phosphine transition metal complex. Within this dose range, the anticancer agent can be administered once a day, or can be administered in several divided doses (for example, about 2 to 4 times).
　本発明の抗がん剤組成物が適用されるがんの種類は、特に限定されるものではなく、例えば、悪性黒色腫、悪性リンパ腫、消化器癌、肺癌、食道癌、胃癌、大腸癌、直腸癌、結腸癌、尿管腫瘍、胆嚢癌、胆管癌、胆道癌、乳癌、肝臓癌、膵臓癌、睾丸腫瘍、上顎癌、舌癌、口唇癌、口腔癌、咽頭癌、喉頭癌、卵巣癌、子宮癌、前立腺癌、甲状腺癌、脳腫瘍、カポジ肉腫、血管腫、白血病、真性多血症、神経芽腫、網膜芽腫、骨髄腫、膀胱腫、肉腫、骨肉腫、筋肉腫、皮膚癌、基底細胞癌、皮膚付属器癌、皮膚転移癌、皮膚黒色腫等が挙げられる。更に、悪性腫瘍ばかりでなく良性腫瘍にも適用され得る。また、本発明の抗がん剤組成物は、がん転移を抑制するために使用されることができ、特に、術後のがん転移抑制剤としても有用である。 The type of cancer to which the anticancer composition of the present invention is applied is not particularly limited, and examples thereof include malignant melanoma, malignant lymphoma, digestive organ cancer, lung cancer, esophageal cancer, stomach cancer, colon cancer, Rectal cancer, colon cancer, ureteral tumor, gallbladder cancer, bile duct cancer, biliary tract cancer, breast cancer, liver cancer, pancreatic cancer, testicular cancer, maxillary cancer, tongue cancer, lip cancer, oral cancer, pharyngeal cancer, laryngeal cancer, ovarian cancer , Uterine cancer, prostate cancer, thyroid cancer, brain tumor, Kaposi's sarcoma, hemangioma, leukemia, polycythemia vera, neuroblastoma, retinoblastoma, myeloma, cystoma, sarcoma, osteosarcoma, muscle tumor, skin cancer, Examples include basal cell cancer, skin appendage cancer, skin metastatic cancer, cutaneous melanoma, and the like. Furthermore, it can be applied not only to malignant tumors but also to benign tumors. Moreover, the anticancer agent composition of the present invention can be used for suppressing cancer metastasis, and is particularly useful as a postoperative cancer metastasis inhibitor.
　本発明の抗がん剤組成物の使用においては、既知の化学療法、外科的治療法、放射線療法、温熱療法や免疫療法などと組み合わせることもできる。 In the use of the anticancer composition of the present invention, it can be combined with known chemotherapy, surgical treatment, radiation therapy, hyperthermia, immunotherapy and the like.
　以下、本発明を実施例により詳細に説明するが本発明はこれらの実施例に限定されるものではない。 Hereinafter, the present invention will be described in detail by way of examples, but the present invention is not limited to these examples.
（ホスフィン金錯体試料の調製）
＜ｔｅｒｔ－ブチルメチルホスフィン－ボラン（７ａ）の合成＞
　下記反応式（１１）に従って、ｔｅｒｔ－ブチルメチルホスフィン－ボラン（７ａ）の合成を行った。
Figure JPOXMLDOC01-appb-C000012
 　ｔｅｒｔ－ブチル（ヒドロキシメチル）メチルホスフィン－ボラン（１０）（１．７８ｇ、１２．０ｍｍｏｌ）を７２ｍＬのアセトンに溶解し、アセトン溶液を得た。次いで、水酸化カリウム（１３．５ｇ、２４０ｍｍｏｌ）、過硫酸カリウム（１９．４ｇ、７２．０ｍｍｏｌ）及び三塩化ルテニウム三水和物（６２４ｍｇ、２．４ｍｍｏｌ）を１５０ｍＬの水に溶解した水溶液を調製し、該水溶液を激しく撹拌した状態で、０℃で、前記アセトン溶液を徐々に添加した。２時間経過後、反応混合液を３Ｍの塩酸で中和し、エーテルで３回抽出した。有機相を飽和食塩水で洗浄し、硫酸ナトリウムで脱水した。ロータリーエバポレータで溶媒を室温下で除去し、シリカゲルのカラムクロマトグラフィー（移動相：ペンタン／エーテル＝８／１）で残渣を精製して、ｔｅｒｔ－ブチルメチルホスフィン－ボラン（７ａ）を得た。収量は２．２７ｇ、収率は８０％であった。 (Preparation of phosphine gold complex sample)
<Synthesis of tert-butylmethylphosphine-borane (7a)>
According to the following reaction formula (11), tert-butylmethylphosphine-borane (7a) was synthesized.
Figure JPOXMLDOC01-appb-C000012
 Tert-butyl (hydroxymethyl) methylphosphine-borane (10) (1.78 g, 12.0 mmol) was dissolved in 72 mL of acetone to obtain an acetone solution. Next, an aqueous solution prepared by dissolving potassium hydroxide (13.5 g, 240 mmol), potassium persulfate (19.4 g, 72.0 mmol) and ruthenium trichloride trihydrate (624 mg, 2.4 mmol) in 150 mL of water was prepared. Then, the acetone solution was gradually added at 0 ° C. while vigorously stirring the aqueous solution. After 2 hours, the reaction mixture was neutralized with 3M hydrochloric acid and extracted three times with ether. The organic phase was washed with saturated brine and dried over sodium sulfate. The solvent was removed with a rotary evaporator at room temperature, and the residue was purified by column chromatography on silica gel (mobile phase: pentane / ether = 8/1) to obtain tert-butylmethylphosphine-borane (7a). The yield was 2.27 g, and the yield was 80%.
＜２，３－ビス（ｔｅｒｔ－ブチルメチルホスフィノ）キノキサリン（３ａ）の合成＞
　下記反応式（１２）に従って、２，３－ビス（ｔｅｒｔ－ブチルメチルホスフィノ）キノキサリン（３ａ）を合成した。
Figure JPOXMLDOC01-appb-C000013
 　２３６ｍｇ（２．０ｍｍｏｌ）のｔｅｒｔ－ブチルメチルホスフィン－ボラン（７ａ）を４ｍＬのテトラヒドロフランに溶解して溶液を得た。この溶液を液体窒素で－７８℃に冷却し、そこに、ｎ－ブチルリチウムのヘキサン溶液（１．６Ｍ）を１．２５ｍＬ滴下した。１５分経過後、１３３ｍｇ（０．６７ｍｍｏｌ）の２，３－ジクロロキノキサリン（６ａ）（関東化学社製）を４ｍＬのテトラヒドロフランに溶解させ、得られた溶液を、激しく撹拌しながら滴下した。１時間かけて液温を室温（２５℃）にした後、３時間撹拌を行った。次いで、１ｍＬのＴＭＥＤＡを添加して、更に２時間撹拌を継続した。１Ｍの塩酸を添加して反応を終了させ、反応液をヘキサンで抽出した。有機相を１Ｍの塩酸及び飽和食塩水で洗浄し、硫酸ナトリウムで脱水した。溶媒を真空吸引で除去し、シリカゲルのカラムクロマトグラフィー（移動相：ヘキサン／酢酸エチル＝３０／１）で残渣を精製し、橙色の固体物を得た。この固体物を、熱メタノール（１．７ｍＬ）で再結晶した。これにより、２，３－ビス（ｔｅｒｔ－ブチルメチルホスフィノ）キノキサリン（３ａ）の橙色結晶を得た。このときの収率は８０％であった。得られた２，３－ビス（ｔｅｒｔ－ブチルメチルホスフィノ）キノキサリン（３ａ）の物性値は以下の通りであった。
（同定データ）
１Ｈ ＮＭＲ（３９５．７５ＭＨｚ，ＣＤＣｌ３）：β １．００－１．０３（ｍ，１８Ｈ）、１．４２－１．４４（ｍ，６Ｈ）、７．７０－７．７４（ｍ，２Ｈ）、８．０８－８．１２（ｍ，２Ｈ）；
１３Ｃ ＮＭＲ（９９．４５ＭＨｚ，ＣＤＣｌ３）：β ４．７７（ｔ，Ｊ＝４．１Ｈｚ）、２７．５９（ｔ，Ｊ＝７．４Ｈｚ）、３１．９０（ｔ，Ｊ＝７．４Ｈｚ）、１２９．５０，１２９．６０，１４１．６３，１６５．１２（ｄｄ，Ｊ＝５．７，２．４Ｈｚ）；
３１Ｐ ＮＭＲ（２０２．３５ＭＨｚ，ＣＤＣｌ３）：β －１７．７（ｓ）；
ＩＲ（ＫＢＲ）２９５０，１４７０，７８０ｃｍ－１；
ＨＲＭＳ（ＦＡＢ）計算値（Ｃ１８Ｈ２９Ｎ２Ｐ２（Ｍ＋＋Ｈ））３３５．１８０９、実測値３３５．１８２６ <Synthesis of 2,3-bis (tert-butylmethylphosphino) quinoxaline (3a)>
2,3-bis (tert-butylmethylphosphino) quinoxaline (3a) was synthesized according to the following reaction formula (12).
Figure JPOXMLDOC01-appb-C000013
 236 mg (2.0 mmol) of tert-butylmethylphosphine-borane (7a) was dissolved in 4 mL of tetrahydrofuran to obtain a solution. This solution was cooled to −78 ° C. with liquid nitrogen, and 1.25 mL of a hexane solution (1.6 M) of n-butyllithium was added dropwise thereto. After 15 minutes, 133 mg (0.67 mmol) of 2,3-dichloroquinoxaline (6a) (manufactured by Kanto Chemical Co.) was dissolved in 4 mL of tetrahydrofuran, and the resulting solution was added dropwise with vigorous stirring. The liquid temperature was brought to room temperature (25 ° C.) over 1 hour, followed by stirring for 3 hours. Then 1 mL TMEDA was added and stirring was continued for another 2 hours. 1M hydrochloric acid was added to terminate the reaction, and the reaction solution was extracted with hexane. The organic phase was washed with 1M hydrochloric acid and saturated brine, and dried over sodium sulfate. The solvent was removed by vacuum suction, and the residue was purified by silica gel column chromatography (mobile phase: hexane / ethyl acetate = 30/1) to obtain an orange solid. This solid was recrystallized with hot methanol (1.7 mL). As a result, orange crystals of 2,3-bis (tert-butylmethylphosphino) quinoxaline (3a) were obtained. The yield at this time was 80%. The physical properties of the obtained 2,3-bis (tert-butylmethylphosphino) quinoxaline (3a) were as follows.
(Identification data)
1 H NMR (395.75 MHz, CDCl 3 ): β 1.00-1.03 (m, 18H), 1.42-1.44 (m, 6H), 7.70-7.74 (m, 2H) ), 8.08-8.12 (m, 2H);
13 C NMR (99.45 MHz, CDCl 3 ): β 4.77 (t, J = 4.1 Hz), 27.59 (t, J = 7.4 Hz), 31.90 (t, J = 7.4 Hz) ) 129.50, 129.60, 141.63, 165.12 (dd, J = 5.7, 2.4 Hz);
31 P NMR (202.35 MHz, CDCl 3 ): β-17.7 (s);
IR (KBR) 2950, 1470, 780 cm −1 ;
HRMS (FAB) calcd (C 18 H 29 N 2 P 2 (M + + H)) 335.1809, Found 335.1826
＜ビス（２，３－ビス（ｔｅｒｔ－ブチルメチルホスフィノ）キノキサリン）金(Ｉ)クロリド（１Ａ）の合成＞
　窒素ガスで置換した２５ｍｌ二口フラスコに、前記合成例１で調製した２，３－ビス（ｔｅｒｔ－ブチルメチルホスフィノ）キノキサリン（３ａ）１．３３ｇ（３．９８ｍｍｏｌ）と脱気したＴＨＦを加えた。ここにテトラブチルアンモニウム金（Ｉ）ジクロリド１．０２ｍｇ（１．９９ｍｍｏｌ）を加え、室温で２０時間撹拌した。沈殿をろ別し、ろ液を乾固した。得られた褐色固体を減圧下で乾燥し、１．４６ｇの下記式（１ａ）で表されるビス（２，３－ビス（ｔｅｒｔ－ブチルメチルホスフィノ）キノキサリン）金(Ｉ)クロリド（１Ａ）を得た（以下、「ホスフィン金錯体試料」と呼ぶ。）。この時の収率は８２％であった。
Figure JPOXMLDOC01-appb-C000014
 （同定データ）
３１Ｐ ＮＭＲ（１２１．５５ＭＨｚ，ＣＤＣｌ３）：８．８（ｓ）
ＭＳ（ＥＳＩ、ＰＯＳ）m/z ８６６ (Ｍ＋－Ｃｌ－) <Synthesis of bis (2,3-bis (tert-butylmethylphosphino) quinoxaline) gold (I) chloride (1A)>
To a 25 ml two-necked flask purged with nitrogen gas was added 1.33 g (3.98 mmol) of 2,3-bis (tert-butylmethylphosphino) quinoxaline (3a) prepared in Synthesis Example 1 and degassed THF. It was. Tetrabutylammonium gold (I) dichloride (1.02 mg, 1.99 mmol) was added thereto, and the mixture was stirred at room temperature for 20 hours. The precipitate was filtered off and the filtrate was dried. The obtained brown solid was dried under reduced pressure, and 1.46 g of bis (2,3-bis (tert-butylmethylphosphino) quinoxaline) gold (I) chloride (1A) represented by the following formula (1a) (Hereinafter referred to as “phosphine gold complex sample”). The yield at this time was 82%.
Figure JPOXMLDOC01-appb-C000014
 (Identification data)
31 P NMR (121.55 MHz, CDCl 3 ): 8.8 (s)
MS (ESI, POS) m / z 866 (M + -Cl -)
｛実施例１～８｝
　表１に示した量と種類のシクロデキストリン化合物を水４ｍｌに加えた。次いで上記で調製したホスフィン金錯体試料１０ｍｇを添加し、２５℃で１６時間攪拌混合して混合処理を行い、各種のシクロデキストリン包接体を得た。
　次いで、２時間静置した後上澄み液を１ｍＬ採取し遠心分離を行った。この上澄み液をさらに０．７ｍｌ採取し遠心分離を２回行い、遠心分離後、上澄み液の０．１００ｍｌを採取し５ｍｌメスフラスコに入れメタノールで５ｍｌにメスアップした。次いで、この溶液を２μＬシリンジで採取しＨＰＬＣで分析してホスフィン金錯体試料の量を測定し、この測定値から溶解度を求めた。その結果を表１に示した。なお、シクロデキストリン化合物を添加しないで、ホスフィン金錯体試料のみのものも同様にして溶解度を求め、ブランクとして表１に結果を併記した。
Figure JPOXMLDOC01-appb-T000015
 {Examples 1 to 8}
The amounts and types of cyclodextrin compounds shown in Table 1 were added to 4 ml of water. Next, 10 mg of the phosphine gold complex sample prepared above was added, mixed by stirring for 16 hours at 25 ° C., and various cyclodextrin inclusion bodies were obtained.
Next, after standing for 2 hours, 1 mL of the supernatant was collected and centrifuged. An additional 0.7 ml of this supernatant was collected and centrifuged twice, and after centrifugation, 0.100 ml of the supernatant was collected and placed in a 5 ml volumetric flask and made up to 5 ml with methanol. Next, this solution was collected with a 2 μL syringe and analyzed by HPLC to measure the amount of the phosphine gold complex sample, and the solubility was determined from the measured value. The results are shown in Table 1. The solubility of the phosphine gold complex sample alone was determined in the same manner without adding the cyclodextrin compound, and the results are shown in Table 1 as blanks.
Figure JPOXMLDOC01-appb-T000015
　表１の結果より、ホスフィン金錯体をシクロデキストリン化合物へ包接させることで、ホスフィン金錯体試料の溶解度が向上することが分かる。特にシクロデキストリン化合物として、β―シクロデキストリンを用いた場合には、少ない添加量でホスフィン金錯体試料の溶解性を劇的に向上させることができることが分かる。 From the results in Table 1, it can be seen that inclusion of the phosphine gold complex in the cyclodextrin compound improves the solubility of the phosphine gold complex sample. In particular, when β-cyclodextrin is used as the cyclodextrin compound, it can be seen that the solubility of the phosphine gold complex sample can be dramatically improved with a small addition amount.
｛実施例９｝
（β－シクロデキストリン包接体試料の調製）
　β―シクロデキストリンの２重量％水溶液５ｍｌを調製し、ホスフィン金錯体試料４０ｍｇを添加し、１６時間、２５℃で攪拌混合することによりβ―シクロデキストリン包接体水溶液を調製した。 {Example 9}
(Preparation of β-cyclodextrin clathrate sample)
5 ml of a 2 wt% aqueous solution of β-cyclodextrin was prepared, 40 mg of a phosphine gold complex sample was added, and the mixture was stirred and mixed at 25 ° C. for 16 hours to prepare a β-cyclodextrin clathrate aqueous solution.
＜抗がん性の評価＞
　β―シクロデキストリン包接体水溶液の腫瘍細胞に対する活性評価を下記のように実施した。また、比較対象としてシスプラチン（比較例１）及びβ―シクロデキストリンを添加しないで水に完全に溶解させたホスフィン金錯体試料（参考例１）についても同様な試験を実施した。
　癌細胞としてＨＬ－６０（ヒト急性骨髄性白血病細胞）を使用し、１０％ウシ胎児血清および１％抗生物質、抗真菌剤を補足したRosewell Park Memorial Institute培地（RPMI1640）中で、５％二酸化炭素雰囲気下、湿潤インキュベーター中、３７℃で培養した。
　細胞はPBSで洗浄し、細胞数を算定後、同じ培地を用いて１×１０６細胞／ｍｌ懸濁液を調製した。滅菌９６ウエルのマイクロプレートに前記の懸濁液を５００００細胞／ウエルの密度となるように加えた。
　次に上記で調製したβ―シクロデキストリン包接体水溶液、ジメチルスルホキシドに完全に溶解させたシスプラチン溶液（比較例１）又はβ―シクロデキストリンを添加しないで水に完全に溶解させたホスフィン金錯体試料（参考例１）を加え、引き続き２４時間インキュベータ中で培養した。
　その後、生存細胞数をMosmann（T.Mosmann, J.Immunnol.Method(1983)）65,55-63)変法により評価した。即ち、テトラゾリウム塩（3,[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide,MTT）溶液を加え、さらに３時間、同条件で培養した。細胞内のミトコンドリアの酵素活性により生成したホルマザン結晶を０．０４ｍｏｌ／ＨＣｌ／イオソプロピルアルコールで溶解し、マイクロプレートリーダー（Bio-Rad 550）を用い、５９５ｎｍの吸光度を測定した。バックグランドを排除するために６３０ｎｍの吸光度を
測定し、実測値から差し引いた。これを生存細胞数として評価し、５０％細胞発育抑制濃度（ＩＣ５０）を算出した。なお、IC50値の算出に当たっては、同様に実施した少なくとも3回以上の実験値の平均値を採用した。この結果を表２に示す。 <Evaluation of anticancer properties>
Evaluation of the activity of the β-cyclodextrin clathrate aqueous solution against tumor cells was performed as follows. Moreover, the same test was implemented also about the phosphine gold complex sample (reference example 1) dissolved completely in water, without adding cisplatin (comparative example 1) and (beta) -cyclodextrin as a comparison object.
Use HL-60 (human acute myeloid leukemia cells) as cancer cells and 5% carbon dioxide in Rosewell Park Memorial Institute medium (RPMI1640) supplemented with 10% fetal bovine serum and 1% antibiotics and antifungals The cells were cultured at 37 ° C. in a humid incubator under an atmosphere.
The cells were washed with PBS, and after counting the number of cells, a 1 × 10 6 cell / ml suspension was prepared using the same medium. The suspension was added to a sterile 96-well microplate to a density of 50000 cells / well.
Next, β-cyclodextrin clathrate aqueous solution prepared above, cisplatin solution completely dissolved in dimethyl sulfoxide (Comparative Example 1), or phosphine gold complex sample completely dissolved in water without adding β-cyclodextrin (Reference Example 1) was added, followed by culturing in an incubator for 24 hours.
Thereafter, the number of viable cells was evaluated by a modified method of Mosmann (T. Mosmann, J. Immunol. Method (1983)) 65, 55-63). That is, a tetrazolium salt (3, [4,5-dimethylthiazole-2-yl] -2,5-diphenyltetrazolium bromide, MTT) solution was added and further cultured for 3 hours under the same conditions. Formazan crystals generated by intracellular mitochondrial enzyme activity were dissolved in 0.04 mol / HCl / iosopropyl alcohol, and absorbance at 595 nm was measured using a microplate reader (Bio-Rad 550). In order to exclude the background, the absorbance at 630 nm was measured and subtracted from the actual measurement value. This was evaluated as the number of viable cells, and the 50% cell growth inhibitory concentration (IC 50 ) was calculated. In calculating the IC 50 value, an average value of at least three experimental values carried out in the same manner was adopted. The results are shown in Table 2.
Figure JPOXMLDOC01-appb-T000016
Figure JPOXMLDOC01-appb-T000016
　表２の結果より、β―シクロデキストリン包接体は、シスプラチンよりも高い抗がん活性を有し、また、ホスフィン金錯体をβ―シクロデキストリン包接体として用いても、ホスフィン金錯体自体の高い抗がん活性をそのまま保持していることが分かる。 From the results in Table 2, β-cyclodextrin inclusion complex has higher anticancer activity than cisplatin, and even if phosphine gold complex is used as β-cyclodextrin inclusion complex, It can be seen that the high anticancer activity is maintained as it is.
　本発明の抗がん剤組成物によれば、溶解性に優れ、高い抗がん活性を有しつつ、毒性の低い抗がん剤組成物が提供される。 According to the anticancer agent composition of the present invention, an anticancer agent composition having excellent solubility and high anticancer activity while having low toxicity is provided.

Claims (7)

  1. 　下記一般式（１）で表されるホスフィン遷移金属錯体と、シクロデキストリン化合物とを含有することを特徴とする抗がん剤組成物。
    Figure JPOXMLDOC01-appb-C000001
     （式中、Ｒ１及びＲ２は、直鎖状若しくは分岐鎖状のアルキル基、シクロアルキル基、置換基を有するシクロアルキル基、アダマンチル基、フェニル基又は置換基を有するフェニル基を示し、炭素数が１～１０であり、同一の基であっても異なる基であってもよい。Ｒ３及びＲ４は、水素原子又は直鎖状若しくは分岐鎖状のアルキル基を示し、炭素数が１～６であり、同一の基であっても異なる基であってもよい。Ｒ３及びＲ４は、互いに結合して飽和又は不飽和の環を形成していてもよく、該飽和又は不飽和の環は、置換基を有していてもよい。Ｍは、金、銅及び銀の群から選ばれる遷移金属原子を示す。Ｘ－は、アニオンを示す。）     An anticancer agent composition comprising a phosphine transition metal complex represented by the following general formula (1) and a cyclodextrin compound.
    Figure JPOXMLDOC01-appb-C000001
     (Wherein R 1 and R 2 represent a linear or branched alkyl group, a cycloalkyl group, a cycloalkyl group having a substituent, an adamantyl group, a phenyl group, or a phenyl group having a substituent; The number may be the same or different, and R 3 and R 4 each represents a hydrogen atom or a linear or branched alkyl group and has 1 carbon atom. And may be the same group or different groups, and R 3 and R 4 may be bonded to each other to form a saturated or unsaturated ring, and the saturated or unsaturated group (The ring in (1) may have a substituent. M represents a transition metal atom selected from the group consisting of gold, copper and silver, and X − represents an anion.)
  2. 　ホスフィン遷移金属錯体が下記一般式（２）で表されることを特徴とする請求項１記載の抗がん剤組成物。
    Figure JPOXMLDOC01-appb-C000002
     （式中、Ｒ１及びＲ２は、直鎖状若しくは分岐鎖状のアルキル基、シクロアルキル基、置換基を有するシクロアルキル基、アダマンチル基、フェニル基又は置換基を有するフェニル基を示し、炭素数が１～１０であり、同一の基であっても異なる基であってもよい。Ｒ５は、一価の置換基を示す。ｎは、０～４の整数を示す。Ｍは、金、銅及び銀の群から選ばれる遷移金属原子を示す。Ｘ－は、アニオンを示す。）     The anticancer agent composition according to claim 1, wherein the phosphine transition metal complex is represented by the following general formula (2).
    Figure JPOXMLDOC01-appb-C000002
     (Wherein R 1 and R 2 represent a linear or branched alkyl group, a cycloalkyl group, a cycloalkyl group having a substituent, an adamantyl group, a phenyl group, or a phenyl group having a substituent; The number may be the same or different, R 5 represents a monovalent substituent, n represents an integer of 0 to 4, M is gold , Represents a transition metal atom selected from the group consisting of copper and silver, and X − represents an anion.)
  3. 　ホスフィン遷移金属錯体の式中のＲ１がｔ－ブチル基であり、Ｒ２がメチル基であることを特徴とする請求項１又は２のいずれか１項記載の抗がん剤組成物。 3. The anticancer agent composition according to claim 1, wherein R 1 in the formula of the phosphine transition metal complex is a t-butyl group and R 2 is a methyl group.
  4. 　ホスフィン遷移金属錯体の式中のＭが金原子であることを特徴とする請求項１乃至３のいずれか１項記載の抗がん剤組成物。 The anticancer agent composition according to any one of claims 1 to 3, wherein M in the formula of the phosphine transition metal complex is a gold atom.
  5. 　シクロデキストリン化合物がα―シクロデキストリン、β－シクロデキストリン及びγ－シクロデキストリンであることを特徴とする請求項１乃至４のいずれか１項記載の抗がん剤組成物。 The anticancer agent composition according to any one of claims 1 to 4, wherein the cyclodextrin compound is α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin.
  6. 　シクロデキストリン化合物がβ－シクロデキストリンであることを特徴とする請求項１乃至４のいずれか１項記載の抗がん剤組成物。 The anticancer agent composition according to any one of claims 1 to 4, wherein the cyclodextrin compound is β-cyclodextrin.
  7. 　シクロデキストリン化合物１００重量部に対してホスフィン遷移金属錯体１０～７５重量部を含有することを特徴とする請求項１乃至６記載のいずれか１項記載の抗がん剤組成物。
     
          The anticancer agent composition according to any one of claims 1 to 6, which comprises 10 to 75 parts by weight of a phosphine transition metal complex with respect to 100 parts by weight of the cyclodextrin compound.

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