JPWO2011125911A1 - Metal complex and anticancer agent containing this as active ingredient - Google Patents

Abstract

The present invention provides a compound having reduced side effects (particularly nephrotoxicity) and excellent anticancer activity as compared with conventional anticancer agents such as cisplatin. A metal complex represented by the following chemical formula 1: or a metal complex represented by the following chemical formula 2: all symbols in the formula are defined in the specification. [Selection] Figure 16

Description

  The present invention relates to a metal complex and an anticancer agent containing this as an active ingredient.

  Cisplatin classified into “platinum preparation (platinum preparation)” is known as an anticancer agent containing a metal complex as an active ingredient. Cisplatin has a structure as a platinum complex as represented by the following chemical formula.

  When this cisplatin is administered, the chlorine atom is eliminated and the platinum atom binds to the N-7 position of guanine or adenine, which is a base of DNA contained in cancer cells. As a result, a cross-link is formed in the DNA strand, and as a result of inhibiting DNA replication, the division and proliferation of cancer cells are suppressed, and the cancer cells are killed.

  Cisplatin was discovered in 1965 by Dr. Rosenberg of the United States as an antibacterial drug that suppresses bacterial growth, and after its antitumor effect was confirmed, it was used for cancer treatment. In Japan, indications for cisplatin are testicular tumor, bladder cancer, renal pelvis / ureter tumor, prostate cancer, ovarian cancer, head and neck cancer, non-small cell lung cancer, esophageal cancer, cervical cancer, neuroblastoma Cellular, gastric cancer, small cell lung cancer, osteosarcoma, germ cell tumor, malignant lymphoma, and so on. In addition, the global market for platinum products is expanding rapidly, and until several years ago, the market grew at a rate of about 20% per year. Although the rate has slowed in recent years, the market for platinum products is still expanding.

  Thus, while cisplatin has a wide range of tumor shrinking effects, it also has a characteristic of showing severe side effects. The most serious side effect is an impaired renal function such as renal failure due to strong nephrotoxicity, which is regarded as a major problem in administration. Since such kidney damage is likely to occur when urine volume decreases, measures such as reducing nephrotoxicity by ingesting water by infusion or increasing urine volume using diuretics can be taken. is necessary. In addition, it is known that gastrointestinal symptoms such as nausea / vomiting and anorexia seen in many patients are expressed considerably more strongly than other anticancer agents. At present, such digestive symptoms are mainly dealt with by using antiemetics together.

  By the way, as one of the platinum preparations as described above, Patent Document 1 discloses various cationic or anionic compounds. The compound described in Patent Document 1 is mainly directed to the treatment of bone-related diseases such as bone cancer.

Special Table 2007-521257

  Although a certain anticancer effect is recognized also about the compound of patent document 1, the effect was not yet satisfactory. In addition, Patent Document 1 does not evaluate the nephrotoxicity of the disclosed compound, and the possibility as a means for solving the problem of side effects inherent in conventional platinum preparations such as cisplatin is unknown. is there.

  Therefore, an object of the present invention is to provide a compound having reduced side effects (particularly nephrotoxicity) and excellent anticancer activity as compared with conventional anticancer agents such as cisplatin.

  The present inventor has intensively studied in view of the above-described problems in the prior art. As a result, it has been found that a certain metal complex containing a noble metal such as platinum (Pt) or palladium (Pd) can solve the above problems, and has completed the present invention.

  That is, according to the first aspect of the present invention, a metal complex represented by the following chemical formula 1 or a salt thereof:

Is provided.

  According to the second embodiment of the present invention, a metal complex represented by the following chemical formula 2 or a salt thereof:

Is provided. In addition, the code | symbol in the said Chemical formula 1 and Chemical formula 2 is demonstrated in detail below.

  ADVANTAGE OF THE INVENTION According to this invention, the side effect (especially nephrotoxicity) is reduced compared with conventional anticancer agents, such as cisplatin, and the compound excellent in anticancer activity is provided.

Obtained by evaluation using cultured human cancer cells panel is a diagram showing a fingerprint for synthesized Pt (5-MP) (AtC3 ) Cl 2 in Example 1-5. Of the fingerprints for Pt (5-MP) (AtC3) Cl ₂ obtained by evaluation using a human cultured cancer cell panel, the “GI50” graph is a known platinum complex anticancer agent. It is the figure shown side by side with the "GI50" graph of cisplatin, carboplatin, and oxaliplatin. It is a graph which shows the result of having evaluated the nephrotoxicity of Pt (5-MP) (AtC3) (Example 1-5) in "Nephrotoxicity evaluation" of an Example. In "nephrotoxicity Evaluation" in _{Example, (Pt (NH 3) 2} · Pt (dach)) - is a graph showing the results of evaluation of the renal toxicity of IP ₆ (Example 2-2). In "nephrotoxicity Evaluation" in Example is a graph showing the results of evaluation of the renal toxicity of _{Pt (Pt (dach) -IP 6} ) 2 ( Example 2-3). It is a graph which shows the result of having evaluated the nephrotoxicity of cisplatin in "Nephrotoxicity evaluation" of an Example. In the “nephrotoxicity evaluation” in the examples, the complex of the present invention was administered to mice, and the mean value of blood glucose values ± standard measured for each sample administration group as compared with the cisplatin administration group and the control group It is a graph which shows a deviation (relative value). In the “nephrotoxicity evaluation” in the examples, the mean value of blood BUN values measured for each sample administration group as compared with the cisplatin administration group and the control group administered to the mice according to the complex of the present invention ± standard It is a graph which shows a deviation (relative value). In the “nephrotoxicity evaluation” in the examples, the mean value of blood Crea values ± standard measured by administering the complex of the present invention to mice and comparing each sample administration group as compared with the cisplatin administration group and the control group It is a graph which shows a deviation (relative value). In the "nephrotoxicity evaluation" of the examples, the complex of the present invention was administered to mice, and the blood hematocrit (Hct) value measured for each sample administration group as compared with the cisplatin administration group and the control group. It is a graph which shows an average value +/- standard deviation (relative value). In the `` nephrotoxicity evaluation '' of the examples, the average of blood hemoglobin (Hb) value measured for each sample administration group as compared with the cisplatin administration group and the control group after administering the complex of the present invention to mice. It is a graph which shows value +/- standard deviation (relative value). Comparison of changes in mean tumor volume ratio of bile cancer mice treated with Pt (5-MP) (AtC3) Cl <sub>2</sub> compared to control group in “Evaluation of anti-cancer effect in vivo (cationic complex)” in Example It is a graph shown. In the “Evaluation of anti-cancer effect in vivo (cationic complex)” in the Examples, the change in the average body weight ratio of the bile cancer mice administered with Pt (5-MP) (AtC3) Cl <sub>2</sub> was compared with the control group. It is a graph shown. Changes in mean tumor volume ratio of bile cancer mice treated with cisplatin or Pd (5-MP) (AtC3) (NO <sub>3</sub> ) <sub>2</sub> in “Evaluation of anticancer effect in vivo (cationic complex)” in Examples Is a graph showing the comparison with the control group. In "Evaluation of anti-cancer effect in vivo (cationic complex)" in the examples, the change in the average body weight ratio of bile cancer mice administered with cisplatin or Pd (5-MP) (AtC3) (NO <sub>3</sub> ) <sub>2</sub> It is a graph shown in comparison with a control group. In the “Evaluation of anti-cancer effect in vivo (anion complex)” in the Examples, the change in the average tumor volume ratio of bile cancer mice administered with cisplatin or Pt (Pt (dach) -IP <sub>6</sub> ) <sub>2</sub> was controlled. It is a graph shown in comparison with. In the `` in vivo evaluation using rat bone metastasis model '' in the examples, the average tumor volume ratio of the cisplatin 33 μmol / kg once-administered group and the Pt (Pt (dach) -IP ₆ ) ₂ 33 μmol / kg once-administered group It is a graph which shows a change compared with a control group. In the “in vivo evaluation using rat bone metastasis model” in the examples, cisplatin 8.25 μmol / kg administered once (day 1 administration) and Pt (Pt (dach) -IP ₆ ) ₂ 8.25 μmol / kg administered twice 3 is a graph showing changes in the average tumor volume ratio of the administration group (administration on days 1 and 8) compared with the control group. In the “in vivo evaluation using rat bone metastasis model” in the examples, cisplatin 8.25 μmol / kg administered once (day 1 administration) and Pt (Pt (dach) -IP ₆ ) ₂ 8.25 μmol / kg administered twice It is a graph which shows the change of the right-and-left foot ratio (average value of a median value) of an administration group (the 1st and 8th day administration) compared with a control group. In the “in vivo evaluation using rat bone metastasis model” in the examples, cisplatin 8.25 μmol / kg administered once (day 1 administration) and Pt (Pt (dach) -IP ₆ ) ₂ 8.25 μmol / kg administered twice It is a graph which shows the change of the body weight (average value) of an administration group (the 1st and 8th day administration) compared with a control group.

  Hereinafter, specific embodiments for carrying out the present invention will be described in detail, but the technical scope of the present invention is not limited to the following specific embodiments.

[First Form: Cation Complex]
A first aspect of the present invention is a metal complex represented by the following chemical formula 1 or a salt thereof:

It is. The metal complex of the first form of the present invention may be in the form of a salt. In this case, the salt of the complex is formed from a cation complex and an anion as a counter ion. Therefore, the metal complex of the first aspect of the present invention may be referred to as “cationic complex”. Moreover, it may be named generically as a "metal complex" including the form of a salt.

  In Chemical Formula 1, M is Pt or Pd. The metal complex (or a salt thereof) of the present embodiment exhibits an excellent anticancer effect by containing platinum (Pt) or palladium (Pd) which are noble metals. Note that M is preferably Pt from the viewpoint that the side effect is less when it is less susceptible to substitution reaction with biological substances such as proteins in the body.

In Chemical Formula 1, R _{1 to} R ₁₇ are each independently a hydrogen atom, an alkyl group having 1 to 30 carbon atoms, a cycloalkyl group having 3 to 30 carbon atoms, an alkenyl group having 2 to 30 carbon atoms, or 3 carbon atoms. -30 cycloalkenyl group, C2-C30 alkynyl group, C7-C30 aralkyl group, C7-C30 aralkenyl group, C7-C30 aralkynyl group, C6-C30 aryl Group, halogen atom, haloalkyl group having 1 to 30 carbon atoms, haloalkenyl group having 2 to 30 carbon atoms, haloalkynyl group having 2 to 30 carbon atoms, haloaryl group having 6 to 30 carbon atoms, alkoxy having 1 to 30 carbon atoms Group, aryloxy group having 6 to 30 carbon atoms, hydroxy group, amino group, alkylamino group having 1 to 30 carbon atoms, arylamino group having 6 to 30 carbon atoms, cyano group, or nitro group A group.

  Examples of the alkyl group having 1 to 30 carbon atoms include, for example, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, t-butyl group, and n-pentyl. Group, isopentyl group, neo-pentyl group, 1,2-dimethylpropyl group, n-hexyl group, cyclo-hexyl group, 1,3-dimethylbutyl group, 1-isopropylpropyl group, 1,2-dimethylbutyl group, n-heptyl group, 1,4-dimethylpentyl group, 2-methyl-1-isopropylpropyl group, 1-ethyl-3-methylbutyl group, n-octyl group, 2-ethylhexyl group, 3-methyl-1-isopropylbutyl Group, 2-methyl-1-isopropyl group, 1-t-butyl-2-methylpropyl group, n-nonyl group, 3,5,5-trimethylhexyl And the like.

  Examples of the cycloalkyl group having 3 to 30 carbon atoms include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, and a cyclooctyl group.

  Examples of the alkenyl group having 2 to 30 carbon atoms include a vinyl group, an allyl group, a propenyl group, an isopropenyl group, a 2-methyl-1-propenyl group, a 2-methylallyl group, and a 2-butenyl group.

  Examples of the C3-C30 cycloalkenyl group include, for example, a cyclopropenyl group, a cyclobutenyl group, a 2-cyclopenten-1-yl group, a 2-cyclohexen-1-yl group, and a 3-cyclohexen-1-yl group. Can be mentioned.

  As an example of a C2-C30 alkynyl group, an ethynyl group, 2-propynyl group, and 2-butynyl group are mentioned, for example.

  Examples of the aralkyl group having 7 to 30 carbon atoms include a benzyl group, a phenethyl group, and a diphenylmethyl group.

  Examples of the aralkenyl group having 7 to 30 carbon atoms include a styryl group, 2-phenyl-1-propenyl group, 3-phenyl-2-butenyl group, and 2-naphthylethenyl group.

  Examples of the aralkynyl group having 7 to 30 carbon atoms include a 2-phenylethynyl group and a 2-naphthylethynyl group.

  Examples of the aryl group having 6 to 30 carbon atoms include a phenyl group, a tolyl group, a xylyl group, an ethylphenyl group, a naphthyl group, and an anthranyl group.

  The halogen atom is fluorine, chlorine, bromine or iodine.

  Examples of the haloalkyl group having 1 to 30 carbon atoms include, for example, a trifluoromethyl group, a difluoromethyl group, a trichloromethyl group, a dichloromethyl group, a fluoromethyl group, a chloromethyl group, an iodomethyl group, a bromomethyl group, and a pentafluoroethyl group. And pentachloroethyl group.

  Examples of the haloalkenyl group having 2 to 30 carbon atoms include, for example, 2,2-difluoroethenyl group, 2,2-dichloroethenyl group, 3-chloro-2-allyl group, 3,3-dichloro-2- Examples include allyl group and 2,3-dibromo-2-allyl.

  Examples of the haloalkynyl group having 2 to 30 carbon atoms include a 3-chloro-2-propynyl group, a 1,3-dichloro-2-propynyl group, and a 1,3-dibromo-2-propynyl group.

  As an example of a C6-C30 haloaryl group, a chlorophenyl group (for example, 4-chlorophenyl group), a bromophenyl group, and a fluorophenyl group are mentioned, for example.

  Examples of the alkoxy group having 1 to 30 carbon atoms include methoxy group, ethoxy group, propoxy group, butoxy group, hexyloxy group, octyloxy group, and 2-ethylhexyloxy.

  Examples of the aryloxy group having 6 to 30 carbon atoms include a phenyloxy group, a naphthyloxy group, and a biphenyloxy group.

  Examples of the alkylamino group having 1 to 30 carbon atoms include a methylamino group, an ethylamino group, a hexylamino group, and a dodecylamino group.

  Examples of the arylamino group having 6 to 30 carbon atoms include a phenylamino group, a naphthylamino group, a benzylamino group, an indanylamino group, and an indenylamino group.

In a preferred embodiment of the present invention, the anthracene ring is unsubstituted and the phenanthroline ring has a substituent only at the 5-position. In other words, R _{1 to} R ₃ and R _{5 to} R ₁₇ are hydrogen atoms, and R ₄ is a group other than a hydrogen atom. In this case, the substituent (R ₄ ) at the 5-position of the phenanthroline ring is preferably an alkyl group having 1 to 30 carbon atoms (in particular, an alkyl group having 1 to 6 carbon atoms, more preferably 1 carbon atom). An alkyl group having 3 to 3 carbon atoms, a halogen atom (especially preferably a chlorine atom), an alkoxy group having 1 to 30 carbon atoms (particularly preferably an alkoxy group having 1 to 6 carbon atoms, more preferably carbon Or a nitro group. In a more preferred embodiment, R _{1 to} R ₃ and R _{5 to} R ₁₇ are hydrogen atoms, and R ₄ is a methyl group, a nitro group, a chlorine atom, or a methoxy group. That is, a preferred specific example of the cation complex (or salt thereof) of this embodiment is represented by any one of the following chemical formulas 1a to 1d.

  Among these, from the viewpoint of having high anticancer activity, the complex represented by Chemical Formula 1a or Chemical Formula 1b is preferable, and the complex represented by Chemical Formula 1a is most preferable.

  In the cation complexes represented by the chemical formulas 1a to 1d described above, stereoisomers may exist depending on the coordination direction of the ligand with respect to the central metal M. This is why some of the coordination bonds are represented by wavy lines (~~~) in the chemical structure described above, and more specifically, both of the two stereoisomers represented by the following chemical formulas. (In the following chemical formula, the case where the central metal M is Pt is described as an example).

  As in the examples described later, when producing a cation complex in which such a stereoisomer may exist, the product may contain the above-described two kinds of steric compounds unless they are selectively produced. It is usually obtained as an equivalent mixture of isomers. In this case, examples of means for purifying only one of the obtained mixture of stereoisomers include a crystallization method and a chromatography method. In the present invention, even in the form of a mixture that has not been subjected to such purification treatment, or in the form of only one isomer after being subjected to purification treatment, both are claimed. It is intended to be included in the technical scope of the described invention.

  In another preferred embodiment of the present invention, an even number of identical alkyl groups having 1 to 30 carbon atoms are bonded to symmetrical positions in the phenanthroline ring in Formula 1. Examples of the symmetrical position in the phenanthroline ring to which an even number of alkyl groups can be bonded include, for example, in the case of two alkyl groups, positions 5, 6 and 2, 9 and the like. Moreover, when there are four alkyl groups, the 3, 4, 7, and 8 positions are listed. In this case, the alkyl group having 1 to 30 carbon atoms is preferably an alkyl group having 1 to 6 carbon atoms, more preferably an alkyl group having 1 to 3 carbon atoms, and most preferably a methyl group. That is, a preferred specific example of the cation complex (or a salt thereof) of this embodiment is represented by any one of the following chemical formulas 1e to 1g.

  As described above, the cation complex of the present invention may be in the form of a salt, but there is no particular limitation on the type of counter anion when the cation complex of the present invention is in the form of a salt. Such counter anions include, for example, chloride ions, fluoride ions, bromide ions, iodide ions and other halide ions, nitrate ions, sulfate ions, phosphate ions, acetate ions, carbonate ions, perchlorate ions. Etc.

  There is no restriction | limiting in particular about the manufacturing method of the cation complex of this invention, Manufacture is possible by considering the description of the Example mentioned later and the technical common sense at the time of application of this application.

As an example of the method for producing the cation complex of the present invention, a 1,10-phenanthroline derivative (DI) is converted into a noble metal complex such as chloroplatinate or palladium chloride as described in the scheme represented by the following chemical formula 9. (Y ₂ MCl ₄ ; Y is an alkali metal) to form a platinum or palladium complex, and the resulting complex is reacted with a diamine derivative having an anthracene ring represented by “AtC3”. The method of obtaining the cation complex of this invention represented by M (DI) (AtC3) is illustrated (refer also the Example mentioned later).

In this case, by appropriately selecting the 1,10-phenanthroline derivative or type of AtC3 used, it is possible to produce a cationic complex having a desired substituent _(R 1 _{to R 17} in Formula 1). Further, by selecting a noble metal atom (Pt or Pd) of the noble metal complex (Y ₂ MCl ₄ ) to be used, a desired noble metal atom can be introduced into the resulting cation complex. In the above scheme, the noble metal complex as a raw material has a chlorine atom as a ligand. However, the present invention is not limited to such a form. It can be used without

  The obtained cation complex can be appropriately purified by a conventionally known purification means. The same applies to other compounds described later.

[Second form: anion complex]
A second aspect of the present invention is a metal complex represented by the following chemical formula 2 or a salt thereof:

It is. The metal complex of the second form of the present invention may be in the form of a salt. In this case, the salt of the complex is formed from an anion complex and a cation as a counter ion. Therefore, the metal complex of the second form of the present invention may be referred to as “anion complex”. Moreover, it may be named generically as a "metal complex" including the form of a salt. In addition, there is no restriction | limiting in particular about the kind of counter cation in case the anion complex of this invention is a salt form. Examples of such counter cations include alkali metal ions such as sodium ions and potassium ions, alkaline earth metal ions such as calcium ions and magnesium ions, quaternary ammonium ions, transition metal ions such as zinc, and lanthanum. Examples include rare earth metal ions and hydrogen ions.

  In Chemical Formula 2, M is Pt or Pd. The metal complex (or a salt thereof) of the present embodiment exhibits an excellent anticancer effect by containing platinum (Pt) or palladium (Pd) which are noble metals. Note that M is preferably Pt from the viewpoint that the side effect is less when it is less susceptible to substitution reaction with biological substances such as proteins in the body.

  In the chemical formula 2, R is a hydrogen atom, or together with the adjacent R, forms a structure represented by the following chemical formula 7.

  Here, “adjacent R” means “R” bonded to the other nitrogen atom (N) bonded to the metal atom (M) bonded to the nitrogen atom (N) bonded to itself (R). . Therefore, although there are four R groups in Chemical Formula 2, the two R groups at the left end in Chemical Formula 2 are the same as each other, and the two R groups at the right end in Chemical Formula 2 are also the same as each other. . That is, two adjacent Rs may be both hydrogen atoms, or otherwise, together, they form a structure represented by the above chemical formula 7.

  Note that when two adjacent Rs together form the structure represented by the chemical formula 7, the three-dimensional structure in the structure is not particularly limited. In the present invention, the bonds bonded to the cyclohexane ring may be in a cis relationship or in a trans relationship. From the viewpoint of anticancer activity, it is more preferable that they have a trans relationship with each other, and it is particularly preferable that they have 1R and 2R configurations.

  In the chemical formula 2, X may be a structure represented by the following chemical formula 3.

  In this case, the anion complex may have a structure represented by the following chemical formula 2d (see also Example 2-4 described later). The anion complex represented by the following chemical formula 2d is one of the preferred embodiments in the second aspect of the present invention.

  In Formula 2d, all Rs form the structure represented by Formula 7 described above. However, in the anion complex of the present invention, at least two adjacent Rs may be hydrogen atoms, Of course, all R may be hydrogen atoms.

  There is no restriction | limiting in particular about the manufacturing method of the anion complex represented by Chemical formula 2d, and it can manufacture by considering the description of the Example (for example, Example 2-4) mentioned later, and the technical common sense at the time of the application of this application. It is.

  On the other hand, in Chemical Formula 2, X may be a structure represented by the following Chemical Formula 4.

The structure represented by Chemical Formula 4 is a structure derived from myo-inositol 6-phosphate ester. Here, myo-inositol 6-phosphate has a structure represented by the following chemical formula. In the present application, the structure represented by the above chemical formula 4 or a structure corresponding thereto may be described as “IP ₆ ”. In the following chemical formula, the phosphate ester group is described as “—OPO ₃ ”, and each has a divalent negative charge (not shown).

As shown in the above chemical formula 4, in the structure represented by the chemical formula 4, four OPO ₃ groups are bonded to the cyclohexane ring, and the remaining two OPO ₃ groups are used for bonding to the metal atom M through an oxygen atom. It has been. In this embodiment, these six groups may be assigned to any of positions 1 to 6 (see the above position numbers) of myo-inositol 6-phosphate ester. However, in the case where X in Chemical Formula 2 has a structure represented by Chemical Formula 4, a preferred embodiment is that two OPO ₃ groups used for bonding to the metal atom M are 2 of myo-inositol 6-phosphate ester. It is the form arrange | positioned so that it may be located in the position and the 5th position. The anion complex according to such an embodiment may have a structure represented by the following chemical formula 2a (see also Example 2-1 described later). The anion complex represented by the following chemical formula 2a is one of the preferred embodiments in the second aspect of the present invention. In the following chemical formula, the phosphate ester group is described as “—OPO ₃ ”, and each has a divalent negative charge (not shown).

In the chemical formula 2a, all R are hydrogen atoms. However, in the anion complex of the present invention, at least two adjacent Rs may be combined to form the structure represented by the chemical formula 7 described above. Alternatively, all Rs may be combined to form the structure represented by Chemical Formula 7 described above. According to such a configuration, as another preferred embodiment, an anion complex represented by the following chemical formula 2b or chemical formula 2e can also be provided (see Example 2-2 and Example 2-5, which will be described later), respectively. . In the following chemical formula, the phosphate ester group is described as “—OPO ₃ ”, and each has a divalent negative charge (not shown).

In addition, in another preferred embodiment in which X in Chemical Formula 2 has a structure represented by Chemical Formula 4, two OPO ₃ groups used for bonding to metal atom M are myo-inositol hexaphosphate esters. It is the form arrange | positioned so that it may be located in 1st place and 2nd place. The anion complex according to such an embodiment may have a structure represented by the following chemical formula 2f (see also Example 2-6 described later). The anion complex represented by the following chemical formula 2f is one of the preferred embodiments in the second aspect of the present invention. In the following chemical formula, the phosphate ester group is described as “—OPO ₃ ”, and each has a divalent negative charge (not shown).

  There is no restriction | limiting in particular about the manufacturing method of the anion complex represented by Chemical formula 2a, 2b, 2e, 2f, and the Example (for example, Example 2-1, 2-2, 2-5, and 2-6) mentioned later is mentioned. Manufacture is possible by taking into consideration the description and common general knowledge at the time of filing of the present application.

  One feature of the anion complex provided by the present invention is that two or more noble metal atoms (Pt or Pd) are contained in one molecule of the metal complex (that is, a multinuclear complex). In the anion complex of the form described so far, two noble metal atoms are contained in one molecule of the complex (binuclear complex). However, according to another embodiment of the present invention, three noble metal atoms are contained in one molecule. A trinuclear complex into which can be introduced can also be provided.

  In such a form, X in Chemical Formula 2 described above is a structure represented by Chemical Formula 5 below.

  In Chemical Formula 5, M has the same definition as above (that is, Pt or Pd). In Chemical Formula 5, A is a structure represented by Chemical Formula 6 described above.

  Also in this embodiment, as in the above-described chemical formula 4, any of the six groups bonded to the cyclohexane ring in chemical formula 6 is in any of positions 1 to 6 (see the above position numbers) of myo-inositol 6-phosphate ester. It may be assigned. However, in a preferred embodiment where X in Chemical Formula 2 has a structure represented by Chemical Formula 5, two bonds in Chemical Formula 6 are located at the 1-position and 2-position of myo-inositol 6-phosphate ester. It is preferable that they are arranged as described above. The anion complex according to such an embodiment may have a structure represented by the following chemical formula 2c (see also Example 2-3 described later). An anion complex represented by the following chemical formula 2c is also one of preferred embodiments in the second aspect of the present invention. The following chemical formula represents a structure in which two units surrounded by parentheses are coordinated to the metal atom M via two oxygen atoms.

  In Formula 2c, all Rs form the structure represented by Formula 7 described above. However, in the anion complex of the present invention, at least two adjacent Rs may be hydrogen atoms, Of course, all R may be hydrogen atoms.

  There is no restriction | limiting in particular about the manufacturing method of the anion complex represented by Chemical formula 2c, It can manufacture by considering the description of the Example (for example, Example 2-3) mentioned later, and the technical common sense at the time of the application of this application. It is.

  However, for example, when the present inventors examined a method for producing a trinuclear complex represented by the chemical formula 2c, it was found that the trinuclear complex can be produced by a scheme different from the above-described binuclear complex. That is, according to this application, the novel manufacturing method of the trinuclear complex in the 2nd form of this invention can also be provided.

  This production method can be summarized as follows: a metal complex represented by the following chemical formula 8:

A reaction with a halogenated platinum salt or a halogenated palladium salt. Thereby, the trinuclear anion complex represented by Chemical Formula 2 described above is produced.

  In Chemical Formula 8, since the definitions of M and R are as described above, detailed description is omitted here.

Halogenated platinum salt (Y ₂ PtZ ₄ (Y is an alkali metal, Z is a halogen atom)) or halogenated palladium salt (Y ₂ PdZ ₄ (Y is an alkali metal, Z is a halogen atom) (Some) can be appropriately selected depending on the type of noble metal atom desired to be introduced into the resulting trinuclear complex. It is preferable to use a salt having the same noble metal atom as the noble metal atom of the metal complex represented by Chemical Formula 8 described above. Moreover, there is no restriction | limiting in particular about an alkali metal, Potassium, sodium, lithium, a cesium etc. are used, Preferably potassium or sodium is used. The halogen atom is not particularly limited, and fluorine (only Pd), chlorine, bromine and iodine can be used, but chlorine is preferably used. Among them, if it is desired to introduce platinum (Pt), potassium chloroplatinate (K ₂ PtCl ₄ ) is preferably used. If it is desired to introduce palladium (Pd), sodium chloropalladate (Na ₂ PdCl ₄ ) is preferably used.

  As is clear from the description of the above-described reaction formula (and Example 2-3 described later), the metal complex represented by the chemical formula 8 described above is approximately about the halogenated platinum salt or the halogenated palladium salt. It is necessary to use 2 times mole. However, the specific amount of raw material to be used can be appropriately determined in consideration of the order of preparation.

  This reaction is preferably carried out in the presence of a halogen scavenger. In the above reaction, halide ions are produced as a by-product, and when this is combined with hydrogen ions, hydrogen halide is generated. The halogen scavenger is added for the purpose of trapping by-produced halide ions, driving them out of the reaction system as a precipitate, and promptly proceeding with the substitution reaction in the solution. As such a halogen scavenger, for example, a silver salt such as silver nitrate, silver sulfate, or silver oxide, or a method such as heating to expel it into the atmosphere as hydrogen halide can be used. The amount of such a halogen scavenger used is not particularly limited, but preferably an amount of halogen scavenger capable of capturing the total amount of by-produced halide ions is used.

  There is no restriction | limiting in particular also about reaction conditions, It can set suitably. For example, the reaction temperature is usually room temperature to 80 ° C, preferably room temperature to 40 ° C. The reaction time is usually 2 hours to 1 week, preferably several hours to several days.

  Since the metal complex according to the present invention has high anticancer activity as described later, it can be used as an anticancer agent.

  That is, according to this invention, the anticancer agent which contains the metal complex mentioned above as an active ingredient is provided.

  The type of cancer to which the anticancer agent of the present invention is applied is not particularly limited. For example, leukemia, malignant melanoma, malignant lymphoma, gastrointestinal 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 tumor, 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, polycythemia vera, neuroblastoma, retinoblastoma, myeloma, cystoma, sarcoma, osteosarcoma, sarcoma, skin cancer, basal cell carcinoma, skin Examples include adnexal cancer, skin metastatic cancer, and cutaneous melanoma. Moreover, it can be applied not only to malignant tumors but also to benign tumors. In addition, the anticancer agent of the present invention can be used for suppressing cancer metastasis, and is particularly useful as a postoperative cancer metastasis inhibitor.

  In using the anticancer agent of the present invention, the anticancer agent of the present invention can be administered to humans or animals (particularly preferably to humans) in various forms. The administration form of the anticancer agent of the present invention may be oral administration if possible, or parenteral administration such as intravenous, intramuscular, subcutaneous or intradermal injection, rectal administration, transmucosal administration and the like. But you can. Examples of the dosage form suitable for oral administration include tablets, pills, granules, powders, capsules, solutions, suspensions, emulsions, syrups and the like. Examples of the pharmaceutical composition suitable for parenteral administration include solid preparations for external use such as injections, drops, nasal drops, sprays, inhalants, suppositories, or ointments, creams, powder coatings, Examples include liquid coating agents, transdermal absorbents such as patches. Furthermore, examples of the dosage form of the anticancer agent of the present invention include embedding pellets and sustained-release preparations prepared by known techniques.

  A preferable administration form, preparation form, and the like are appropriately selected by a doctor according to the age, sex, constitution, symptom, treatment time, etc. of the patient.

  When the anticancer agent of the present invention is a solid preparation such as a tablet, pill, powder, powder, granule or the like, these solid preparations contain the metal complex according to the present invention as a suitable additive according to a conventional method, For example, lactose, sucrose, D-mannitol, corn starch, synthetic or natural gum, excipients such as crystalline cellulose, starch, hydroxypropylcellulose, hydroxypropylmethylcellulose, gum arabic, gelatin, polyvinylpyrrolidone and other binders, carbo Disintegrants such as carboxymethylcellulose calcium, sodium carboxymethylcellulose, starch, corn starch, sodium alginate, lubricants such as talc, magnesium stearate, sodium stearate, calcium stearate, calcium carbonate, sodium carbonate, calcium phosphate, By appropriately mixing the filler or diluent, such as sodium and the like, it can be manufactured. 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, titanium oxide and the like.

  When the anticancer agent of the present invention is a liquid preparation such as injection, eye drop, nasal drop, inhalant, spray, lotion, syrup, liquid, suspension, emulsion, etc., these liquid preparations Is a metal complex according to the present invention, purified water, a suitable buffer solution such as phosphate buffer, physiological saline solution, physiological salt solution such as Ringer's solution, lock solution, vegetable oil such as cocoa butter, sesame oil, olive oil, etc. , Mineral oil, higher alcohols, higher fatty acids, ethanol and other organic solvents, and if necessary, emulsifiers such as cholesterol, suspending agents such as gum arabic, dispersion aids, wetting agents, polyoxyethylene hydrogenated castor oil , Surfactants such as polyethylene glycol, solubilizers such as sodium phosphate, stabilizers such as sugar, sugar alcohol and albumin, preservatives such as paraben, sodium chloride, glucose, glycerin Isotonic agent, buffer, soothing agent, anti-adsorption agent, moisturizer, antioxidant, colorant, sweetener, flavor, fragrance, etc. It can be prepared as a suspension, liposome or emulsion. In this case, the injection preferably has a physiological pH, and particularly preferably has a pH within the range of 6-8.

  When the anticancer agent of the present invention is a semi-solid preparation such as a lotion, cream or ointment, these semi-solid preparations contain the metal complex according to the present invention as a fat, fatty oil, lanolin, petrolatum, paraffin, wax. , Plasters, resins, plastics, glycols, higher alcohols, glycerin, water, emulsifiers, suspending agents and the like.

  In the anticancer agent of the present invention, the content of the metal complex according to the present invention may vary depending on the dosage form, severity, desired dosage, etc. It is 0.001-80 mass% with respect to the total mass of an adhesive agent, Preferably it is 0.1-50 mass%.

  The dosage of the anticancer agent of the present invention can be appropriately determined by a doctor according to conditions such as the age, sex, weight, symptoms, and administration route of the patient. In general, the amount of active ingredient per day for an adult 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. Such a dose of anticancer agent may be administered once a day, or may be administered divided into several times a day (for example, about 2 to 4 times).

  In using the anticancer agent of the present invention, it may be used in combination with known chemotherapy, surgical treatment, radiation therapy, hyperthermia, immunotherapy and the like.

  The anticancer agent of the present invention exhibits extremely high anticancer properties as shown in the examples described later. In addition, the anticancer agent of the present invention has significantly reduced side effects as compared with conventional platinum preparations in which serious side effects (nephrotoxicity) are a problem. Furthermore, it has been suggested that the mode of action of the anticancer agent of the present invention is also different from existing anticancer agents. Therefore, the anticancer agent of the present invention has a possibility of replacing an existing platinum preparation, and is a very promising new drug candidate.

  In addition, the proteasome inhibitory action was recognized in the metal complex of the present invention as shown in Examples described later. The proteasome is an enzyme that selectively degrades polyubiquitinated intracellular proteins, and plays a central role in the degradation of proteins that control the cell cycle and apoptosis. For this reason, it is considered that this proteasome inhibitory action is involved in at least part of the mechanism by which the complex of the present invention exhibits anticancer properties. In addition, most of the complexes of the present invention inhibit one or more of various cancer-related enzymes such as telomerase, farnesyltransferase, histone deacetylase, and protein kinase. From this, it is considered that the anticancer property of the complex of the present invention is exhibited as a comprehensive action in combination with not only the above-described inhibitory action of the proteasome but also the inhibitory action against other enzymes. . Based on the above findings, the present application also includes a proteasome inhibitor, a telomerase inhibitor, a farnesyltransferase inhibitor, a histone deacetylase inhibitor, or a protein kinase inhibitor, which contains the above-described metal complex of the present invention as an active ingredient. It can also be provided.

Furthermore, among the anion complexes of the present invention, as a characteristic characteristic of those having an IP ₆ structure, the anion complex is more hydroxyapatite (HAP; Ca ₁₀ (PO ₄ ) ₆ (OH) than the conventional platinum preparation. ₂ ) is characterized by a large adsorption equilibrium constant a. Accordingly, among the anion complexes of the present invention, those having an IP ₆ structure alone or complexed with HAP can be targeted to cells affected by bone-related diseases such as bone cancer. It can be used as a system (DDS) formulation. Based on such knowledge, according to the present invention, an anion complex of the present invention having an IP ₆ structure complexed with HAP can also be provided. The anion complex complexed with HAP can be used for the same applications as described above. In particular, it is expected to be useful as a therapeutic agent for bone-related diseases such as bone cancer. The method for producing such a complex is not particularly limited. For example, the complex can be produced by incubating the predetermined anion complex with HAP in an appropriate buffer (for example, HEPES buffer). is there.

  EXAMPLES Hereinafter, although this invention is demonstrated in detail using an Example, the technical scope of this invention is not necessarily restrict | limited only to the form as described in the following Example.

[Synthesis of Cation Complex (M (DI) (AtC3))]
The cation complex of the present invention was synthesized according to the scheme shown in the following chemical formula 9.

  In the structure of the compound represented by Chemical Formula 9, R represents a substituent to the phenanthroline skeleton, A represents an alkali metal, and M represents Pt (II) or Pd (II). Of the target products synthesized in the following examples, those in which a substituent is introduced asymmetrically into the phenanthroline ring (that is, a product in which a substituent is introduced only at the 5-position of the phenanthroline ring) It is obtained as an equal mixture of two kinds of products represented by the following chemical formula 10.

  Reagents used in the synthesis of the cation complex were obtained by the following routes, respectively.

K ₂ PtCl ₂ , Na ₂ PdCl ₄ : purchased from Tanaka Kikinzoku Kogyo Co., Ltd. 5,6-dimethylphenanthroline (5,6-DMP): purchased from Fluka 2,9-dimethylphenanthroline (2,9-DMP), 3 , 4,7,8-tetramethylphenanthroline (3,4,7,8-TMP): purchased from SIGMA 5-nitrophenanthroline (5-NP), 5-methylphenanthroline (5-MP), 5-chlorophenanthroline (5-ClP): purchased from Tokyo Chemical Industry Co., Ltd. 5-methoxyphenanthroline (5-OMP): synthesized according to the literature (Y. Shen, BP Sullivan, Inorg. Chem., 34, 6235-6 (1995)). Dimethyl sulfoxide (DMSO): purchased from Nacalai Tesque, Inc. Acetone: purchased from Wako Pure Chemical Industries, Ltd. [Synthesis Example 1-1: Pt Synthesis of (5,6-DMP) Cl ₂ ]
0.708 g (3.4 mmol) of 5,6-DMP was dissolved in 1 mL of DMSO, and 1.38 g (3.3 mmol) of K ₂ PtCl ₂ was suspended in 24 mL of H ₂ O. After mixing these, the mixture was heated and stirred at 80 ° C. for 3 hours, and then stirred at room temperature for 2 hours. The produced yellow powder was collected by filtration, washed with H ₂ O and dried.

[Synthesis Example 1-2: Synthesis of Pd (5,6-DMP) Cl ₂ ]
0.708 g (3.4 mmol) of 5,6-DMP was dissolved in 1 mL of DMSO, and 0.745 g (3.3 mmol) of Na ₂ PdCl ₄ was suspended in 24 mL of H ₂ O. This was heated and stirred at 80 ° C. for 3 hours, and then stirred at room temperature for 2 hours. The produced yellow powder was collected by filtration, washed with H ₂ O and dried.

[Synthesis Example 1-3: Synthesis of AtC3 · 2HCl]
AtC3 · 2HCl was synthesized by the method described in JP-T-2007-521257 (paragraphs “0079” to “0081”).

Example 1-1 Synthesis of Pt (5,6-DMP) (AtC3) Cl ₂

0.474 g (1 mmol) of Pt (5,6-DMP) Cl ₂ synthesized by the same method as in Synthesis Example 1-1 was suspended in 10 mL of H ₂ O. Next, 0.42 g (1.3 mmol) of AtC3 · 2HCl and 0.16 g (1.5 mmol) of Na ₂ CO ₃ synthesized by the method of Synthesis Example 1-3 described above were added to 10 mL of H ₂ O and ethanol. A solution dissolved in 5 mL of a mixed solvent was added. Then, when the obtained reaction liquid was heated and stirred at 80 ° C. for 3 hours, a yellow solution was obtained. This was immediately filtered while hot, and then concentrated under reduced pressure until the amount of the filtrate became 1 to 2 mL. Acetone was added to the concentrated solution and left in the refrigerator overnight to obtain a yellow powder. This powder was collected by filtration, washed with acetone and dried to obtain the desired product (yield 0.22 g, yield 26%).

Elemental analysis _{C 32 H 32 N 4 PtCl 2} · 6H 2 O
Found: C 45.5%, H 4.83%, N 6.60%
Calculated values: C 45.4%, H 5.20%, N 6.62%

Example 1-2: Pd (5,6-DMP) (AtC3) Synthesis of Cl _2]

Pd (5,6-DMP) Cl ₂ 0.385 g (1 mmol) synthesized by the same method as in Synthesis Example 1-2 described above was suspended in 10 mL of H ₂ O. Next, a solution of 0.42 g (1.3 mmol) of AtC3 · 2HCl and 0.16 g (1.5 mmol) of Na ₂ CO _{3 in} a mixed solvent of 10 mL of H ₂ O and 5 mL of ethanol was added to this suspension. . Then, when the obtained reaction liquid was heated and stirred at 80 ° C. for 3 hours, a yellow solution was obtained. This was immediately filtered while hot, and then concentrated under reduced pressure until the amount of the filtrate became 1 to 2 mL. Acetone was added to the concentrated solution and left in the refrigerator overnight to obtain a yellow powder. This powder was collected by filtration, washed with acetone and dried to obtain the desired product (yield 0.53 g, yield 47.5%).

Elemental analysis _{C 32 H 32 N 4 PdCl 2} · 3H 2 O · 7NaCl
Found: C 34.4%, H 3.54%, N 4.88%
Calculated values: C 34.5%, H 3.41%, N 5.03%

Example 1-3: Pt (2,9-DMP) (AtC3) Synthesis of Cl _2]

0.474 g (1 mmol) of Pt (2,9-DMP) Cl ₂ synthesized by the same method as in Synthesis Example 1-1 was suspended in 10 mL of H ₂ O. Next, a solution of 0.42 g (1.3 mmol) of AtC3 · 2HCl and 0.16 g (1.5 mmol) of Na ₂ CO _{3 in} a mixed solvent of 10 mL of H ₂ O and 5 mL of ethanol was added to this suspension. . Then, when the obtained reaction liquid was heated and stirred at 80 ° C. for 2 hours, a yellow solution was obtained. This was immediately filtered while hot, and then concentrated under reduced pressure until the amount of the filtrate became 1 to 2 mL. Acetone was added to the concentrated solution and left in the refrigerator overnight to obtain a yellow powder. The powder was collected by filtration, washed with acetone and dried to obtain the desired product (yield 0.30 g, yield 38%).

Elemental analysis _{C 32 H 32 N 4 PtCl 2} · 1EtOH
Found: C 51.5%, H 5.37%, N 6.87%
Calculated values: C 52.0%, H 4.85%, N 7.14%

Example 1-4: Pd (2,9-DMP) (AtC3) Synthesis of Cl _2]

Pd (2,9-DMP) Cl ₂ 0.385 g (1 mmol) synthesized by the same method as in Synthesis Example 1-2 described above was suspended in 10 mL of H ₂ O. Next, a solution of 0.42 g (1.3 mmol) of AtC3 · 2HCl and 0.16 g (1.5 mmol) of Na ₂ CO _{3 in} a mixed solvent of 10 mL of H ₂ O and 5 mL of ethanol was added to this suspension. . Thereafter, the obtained reaction solution was heated and stirred at 80 ° C. for 4 hours, and stirred at 40 ° C. overnight, whereby a yellow solution was obtained. This was immediately filtered while hot, and then concentrated under reduced pressure until the amount of the filtrate became 1 to 2 mL. Acetone was added to the concentrated solution and left in the refrigerator overnight to obtain a yellow powder. The powder was collected by filtration, washed with acetone, and dried to obtain the desired product (yield 0.21 g, yield 17%).

Elemental analysis _{C 32 H 32 N 4 PdCl 2} · 4.5H 2 O · 0.5C 3 H 6 O · 8NaCl
Found: C 33.1%, H 3.21%, N 4.08%
Calculated values: C 32.8%, H 3.34%, N 4.56%

Example 1-5: Pt (5-MP) (AtC3) Synthesis of Cl _2]

0.460 g (1 mmol) of Pt (5-MP) Cl ₂ synthesized by the same method as in Synthesis Example 1-1 was suspended in 10 mL of H ₂ O. Next, a solution of 0.42 g (1.3 mmol) of AtC3 · 2HCl and 0.16 g (1.5 mmol) of Na ₂ CO _{3 in} a mixed solvent of 10 mL of H ₂ O and 5 mL of ethanol was added to this suspension. . Then, when the obtained reaction liquid was heated and stirred at 80 ° C. for 3 hours, a yellow solution was obtained. This was immediately filtered while hot, and then concentrated under reduced pressure until the amount of the filtrate became 1 to 2 mL. Acetone was added to the concentrated solution and left overnight in the refrigerator to obtain a black powder. The powder was collected by filtration, washed with acetone, and dried to obtain the desired product (yield 0.45 g, yield 53%).

Elemental analysis _{C 31 H 30 N 4 PtCl 2} · 5H 2 O · 0.5NaCl
Found: C 44.1%, H 4.30%, N 6.26%
Calculated values: C 44.1%, H 4.74%, N 6.64%

Example 1-6: Pt (5-MP) (AtC3) (NO 3) 2 of Synthesis

Pt synthesized by the procedure of Example 1-5 described above (5-MP) (AtC3) a Cl ₂ was dissolved in water (about 1M), ion exchange resins IRA400J ^- was passed through a (NO ₃ form). Acetone was added to a solution obtained by concentrating the filtrate under reduced pressure, and the filtrate was left overnight in a refrigerator to obtain a powder. The powder was collected by filtration, washed with acetone and dried to obtain the desired product.

Elemental analysis _{C 31 H 30 N 6 O 6} Pt · 0.5H 2 O
Actual value: C 47.2%, H 3.86%, N 10.3%
Calculated values: C 47.3%, H 3.94%, N 10.7%

Example 1-7-1: Pd (5-MP) (AtC3) Synthesis of Cl _2]

Pd (5-MP) Cl ₂ 0.371 g (1 mmol) synthesized by the same method as in Synthesis Example 1-2 described above was suspended in 10 mL of H ₂ O. Next, a solution of 0.42 g (1.3 mmol) of AtC3 · 2HCl and 0.16 g (1.5 mmol) of Na ₂ CO _{3 in} a mixed solvent of 10 mL of H ₂ O and 5 mL of ethanol was added to this suspension. . Then, when the obtained reaction liquid was heated and stirred at 80 ° C. for 3 hours, a yellow solution was obtained. This was immediately filtered while hot, and then concentrated under reduced pressure until the amount of the filtrate became 1 to 2 mL. Acetone was added to the concentrated solution and left overnight in the refrigerator to obtain a black powder. The powder was collected by filtration, washed with acetone, and dried to obtain the desired product (yield 0.37 g, yield 39%).

Elemental analysis _{C 31 H 30 N 4 PdCl 2} · 1H 2 O · 5NaCl
Actual value: C 39.2%, H 3.36%, N 5.73%
Calculated values: C 39.3%, H 3.38%, N 5.92%

Example 1-7-2: Pd (5-MP) (AtC3) (NO 3) 2 of Synthesis

Using Pd (5-MP) (AtC3) Cl ₂ synthesized by the same method as in Example 1-7-1 described above as a raw material, the target product was obtained by the same method as in Example 1-6 described above It was.

Elemental analysis _{C 31 H 30 N 6 O 6} Pd · 2H 2 O
Actual value: C 51.68%, H 4.38%, N 11.3%
Calculated values: C 51.35%, H 4.69%, N 11.62%
Example 1-8: Pt (5-NP) (AtC3) Synthesis of Cl _2]

0.484 g (1 mmol) of Pt (5-NP) Cl ₂ synthesized by the same method as in Synthesis Example 1-1 was suspended in 10 mL of H ₂ O. Next, a solution of 0.42 g (1.3 mmol) of AtC3 · 2HCl and 0.16 g (1.5 mmol) of Na ₂ CO _{3 in} a mixed solvent of 10 mL of H ₂ O and 5 mL of ethanol was added to this suspension. . Then, when the obtained reaction liquid was heated and stirred at 80 ° C. for 3 hours, a black solution was obtained. This was immediately filtered while hot, and then concentrated under reduced pressure until the amount of the filtrate became 1 to 2 mL. Acetone was added to the concentrated solution and left overnight in the refrigerator to obtain a black powder. The powder was collected by filtration, washed with acetone, and dried to obtain the desired product (yield 0.17 g, yield 20%).

Elemental analysis _{C 30 H 27 N 5 O 2} PtCl 2 · 6H 2 O
Found: C 41.7%, H 4.38%, N 7.6%
Calculated values: C 41.7%, H 4.52%, N 8.11%

Example 1-9: Pt (5-NP) (AtC3) (NO 3) 2 of Synthesis

Using Pt synthesized in the same manner as in Example 1-8 described above (5-NP) to (AtC3) Cl ₂ as a starting material, the same manner as in Example 1-6 described above, to yield the expected product.

Elemental analysis _{C 30 H 27 N 5 O 2} PtCl 2 · 3C 3 H 6 O
Found: C 45.8%, H 3.93%, N 9.07%
Calculated values: C 44.9%, H 4.31%, N 9.40%

Example 1-10: Pd (5-NP) (AtC3) Synthesis of Cl _2]

0.402 g (1 mmol) of Pd (5-NP) Cl ₂ synthesized by the same method as in Synthesis Example 1-2 described above was suspended in 10 mL of H ₂ O. Next, a solution of 0.42 g (1.3 mmol) of AtC3 · 2HCl and 0.16 g (1.5 mmol) of Na ₂ CO _{3 in} a mixed solvent of 10 mL of H ₂ O and 5 mL of ethanol was added to this suspension. . Then, when the obtained reaction liquid was heated and stirred at 80 ° C. for 3 hours, a black solution was obtained. This was immediately filtered while hot, and then concentrated under reduced pressure until the amount of the filtrate became 1 to 2 mL. Acetone was added to the concentrated solution and left overnight in the refrigerator to obtain a black powder. The powder was collected by filtration, washed with acetone and dried to obtain the desired product (yield 0.27 g, yield 31%).

Elemental analysis _{C 30 H 27 N 5 O 2} PdCl 2 · 1H 2 O · 3.5C 3 H 6 O
Actual value: C 54.2%, H 5.19%, N 7.41%
Calculated values: C 54.7%, H 5.63%, N 7.89%

Example 1-11: Pt (3,4,7,8-TMP) (AtC3) Synthesis of Cl _2]

Pt (3,4,7,8-TMP) Cl ₂ 0.31 g (0.62 mmol) synthesized by the same method as in Synthesis Example 1-1 was suspended in 10 mL of H ₂ O. Next, a solution prepared by dissolving 0.26 g (0.80 mmol) of AtC3 · 2HCl and 0.092 g (0.86 mmol) of Na ₂ CO _{3 in} a mixed solvent of 10 mL of H ₂ O and 5 mL of ethanol was added to the suspension. . Then, when the obtained reaction liquid was heated and stirred at 80 ° C. for 3 hours, a yellow solution was obtained. This was immediately filtered while hot, and then concentrated under reduced pressure until the amount of the filtrate became 1 to 2 mL. Acetone was added to the concentrated solution and left overnight in the refrigerator to obtain a black powder. The powder was collected by filtration, washed with acetone and dried to obtain the desired product (yield 0.16 g, yield 21%).

Elemental analysis C ₃₄ H ₃₆ N ₄ PtCl ₂ · 7H ₂ O · 2NaCl
Actual value: C 40.4%, H 4.60%, N 5.67%
Calculated values: C 40.4%, H 4.96%, N 5.55%

Example 1-12: Pt (3,4,7,8-TMP) (AtC3) (NO 3) 2 of Synthesis

Using Pt (3,4,7,8-TMP) (AtC3) Cl ₂ synthesized by the same method as in Example 1-11 described above as a raw material, The product was obtained.
Elemental analysis _{C 34 H 36 N 6 O 6} Pt · 0.5C 3 H 6 O
Found: C 49.3%, H 5.04%, N 9.41%
Calculated values: C 49.7%, H 4.67%, N 9.80%
Example 1-13: Pd (3,4,7,8-TMP) (AtC3) Synthesis of Cl _2]

Pd (3,4,7,8-TMP) Cl ₂ 0.39 g (0.94 mmol) synthesized by the same method as in Synthesis Example 1-2 described above was suspended in 10 mL of H ₂ O. Next, a solution of 0.39 g (1.2 mmol) of AtC3 · 2HCl and 0.14 g (1.3 mmol) of Na ₂ CO _{3 in} a mixed solvent of 10 mL of H ₂ O and 5 mL of ethanol was added to this suspension. . Then, when the obtained reaction liquid was heated and stirred at 80 ° C. for 3 hours, a black solution was obtained. This was immediately filtered while hot, and then concentrated under reduced pressure until the amount of the filtrate became 1 to 2 mL. Acetone was added to the concentrated solution and left in the refrigerator overnight to obtain a yellow powder. The powder was collected by filtration, washed with acetone and dried to obtain the desired product (yield 0.53 g, yield 62%).

Elemental analysis C ₃₄ H ₃₆ N ₄ PdCl ₂ · 7H ₂ O · 2NaCl
Found: C 44.0%, H 4.95%, N 6.00%
Calculated: C 44.3%, H 5.43%, N 6.08%

Example 1-14: Pt (5-ClP) (AtC3) Synthesis of Cl _2]

Pt (5-ClP) Cl ₂ 0.33 g (0.7 mmol) synthesized by the same method as in Synthesis Example 1-1 was suspended in 10 mL of H ₂ O. Next, a solution of 0.32 g (1.0 mmol) of AtC3 · 2HCl and 0.14 g (1.3 mmol) of Na ₂ CO _{3 in} 10 mL of H ₂ O was added to this suspension. Then, when the obtained reaction liquid was heated and stirred at 80 ° C. for 3 hours, a yellow solution was obtained. This was immediately filtered while hot, and then concentrated under reduced pressure until the amount of the filtrate became 1 to 2 mL. Acetone was added to the concentrated solution and left overnight in the refrigerator to obtain a black powder.

Example 1-15: Pt (5-ClP) (AtC3) (NO 3) 2 of Synthesis

Using Pt synthesized in the same manner as in Example 1-14 described above (5-ClP) a (AtC3) Cl ₂ as a starting material, the same manner as in Example 1-6 described above, to yield the expected product.

Elemental analysis _{C 30 H 27 N 6 O 6} PtCl · 2H 2 O
Found: C 43.3%, H 3.26%, N 9.61%
Calculated values: C 43.2%, H 3.72%, N 10.1%

Example 1-16: Pd (5-ClP) (AtC3) Synthesis of Cl _2]

Pd (5-ClP) Cl ₂ 0.39 g (1.0 mmol) synthesized by the same method as in Synthesis Example 1-2 described above was suspended in 10 mL of H ₂ O. Then, a solution of 0.42 g (1.3 mmol) of AtC3 · 2HCl and 0.16 g (1.5 mmol) of Na ₂ CO _{3 in} 10 mL of H ₂ O was added to this suspension. Then, when the obtained reaction liquid was heated and stirred at 80 ° C. for 3 hours, a yellow solution was obtained. This was immediately filtered while hot, and then concentrated under reduced pressure until the amount of the filtrate became 1 to 2 mL. Acetone was added to the concentrated solution and left in the refrigerator overnight to obtain a yellow powder. The powder was collected by filtration, washed with acetone, and dried to obtain the desired product (yield 0.76 g, yield 89%).

Elemental analysis _{C 30 H 27 N 4 PdCl 2} · 3H 2 O · 3NaCl
Actual value: C 43.1%, H 3.74%, N 5.96%
Calculated values: C 42.4%, H 3.88%, N 6.59%

Example 1-17: Pd (5-ClP) (AtC3) (NO 3) 2 of Synthesis

Using Pd synthesized in the same manner as in Example 1-14 described above (5-ClP) a (AtC3) Cl ₂ as a starting material, the same manner as in Example 1-6 described above, to afford the desired product ( Yield 0.76 g, 89% yield).

Elemental analysis _{C 30 H 27 N 6 O 6} Pd · 1.5H 2 O
Actual value: C 48.91%, H 4.23%, N 10.93%
Calculated: C 48.93%, H 4.78%, N 11.42%

Example 1-18: Pt (5-OMP) (AtC3) Synthesis of Cl _2]

0.476 g (1 mmol) of Pt (5-OMP) Cl ₂ synthesized by the same method as in Synthesis Example 1-1 was suspended in 20 mL of H ₂ O. To this suspension was then added 0.42 g (1.3 mmol) of AtC3 · 2HCl and 0.15 g (1.5 mmol) of Na ₂ CO ₃ . Then, when the obtained reaction liquid was heated and stirred at 80 ° C. for 3 hours, a brown solution was obtained. This was immediately filtered while hot, and then concentrated under reduced pressure until the amount of the filtrate became 1 to 2 mL. Ethanol was added to the concentrated solution to form an ocher precipitate. The precipitate was collected by filtration, washed with ethanol and ether, and dried with a vacuum desiccator. This was distributed to n-butanol / water, the n-butanol layer was removed by an evaporator, and the residue was recrystallized from hexane / ether (9/1) to obtain the desired product (yield 0.26 g, Yield 34%)
Elemental analysis _{C 31 H 32 N 4 O 3} PtCl 2
Found: C 48.1%, H 3.95%, N 7.09%
Calculated values: C 48.1%, H 4.16%, N 7.23%

[Synthesis of anion complexes]
Reagents used in the synthesis of the anion complex were obtained by the following routes, respectively.

K ₂ PtCl _2: Tanaka Kikinzoku purchased from Ltd. myo- inositol 1,2,3,4,5,6-hexaphosphate ester 12 Sodium salt _(IP 6 · 12Na): SIGMA Co. purchased from 1R, 2R- 1,2-cyclohexanediamine (dach): purchased from Tokyo Chemical Industry Co., Ltd. N, N-dimethylacetamide (DMA), AgNO ₃ : purchased from Wako Pure Chemical Industries, Ltd. 28% by mass ammonia water: purchased from Nacalai Tesque, Inc. [Synthesis Example 2-1: Synthesis of cis-Pt (NH ₃ ) ₂ I ₂ ]
Cis-Pt (NH ₃ ) ₂ I ₂ was synthesized by the method described in JP-T-2007-521257 (paragraph “0050”).

[Synthesis Example 2-2: Synthesis of cis-Pt (NH ₃ ) ₂ -IP ₆ ]
Cis-Pt (NH ₃ ) ₂ -IP ₆ was synthesized by the method described in JP-T-2007-521257 (paragraphs “0095” to “0098”).

[Synthesis Example 2-3: Synthesis of cis-Pt (dach) I ₂ ]
Cis-Pt (dach) I ₂ was synthesized by the method described in JP-T-2007-521257 (paragraphs “0057” to “0058”).

[Synthesis Example 2-4: Synthesis of cis-Pt (dach) -IP ₆ ]
Cis-Pt (dach) -IP ₆ was synthesized by the method described in JP-T-2007-521257 (paragraphs “0098” to “0103”).

[Synthesis Example 2-5: Synthesis of cis-Pt (dach) -PP]
Cis-Pt (dach) -PP was synthesized according to the scheme shown in Chemical Formula 11 below.

Ag ₂ SO ₄ (0.315 g, 0.98 mmol) was suspended in H ₂ O (40 mL). To this, 10 mL of dimethylacetamide (DMA) solution in which 0.57 g (1.0 mmol) of cis-Pt (dach) I ₂ synthesized by the method of Synthesis Example 2-3 described above was dissolved was added, and light-shielded. Stir for hours. To the solution from which AgI was removed by filtration, 0.315 g (0.98 mmol) of Ba (OH) ₂ .8H ₂ O was added and stirred for 30 minutes. Pyrophosphate (PP) (1.0 mmol) was added to the solution from which BaSO ₄ was removed by filtration, and stirred for 4 hours. The filtrate was concentrated with an evaporator to obtain a dark brown powder. The precipitated powder was reprecipitated with H ₂ O and acetone to obtain the desired product (yield 0.37 g, yield 77%).

Elemental analysis _{C 6 H 16 N 2 O 7} P 2 Pt · H 2 O
Calculated values: C 14.31%, H 3.61%, N 5.58%
Found: C 14.79%, H 3.60%, N 5.57%
X-ray fluorescence analysis Calculated values: P 2.00, Pt 1.00
Actual value: P 2.00, Pt 0.990

Example _{2-1: (Pt (NH 3)} 2) Synthesis of 2 -IP _6]
(Pt (NH ₃ ) ₂ ) ₂ -IP ₆ which is a binuclear anion complex of the present invention was synthesized according to the scheme shown in the following chemical formula 12.

AgNO ₃ (0.34 g, 2.0 mmol) was dissolved in 40 mL of H ₂ O, and cis-Pt (NH ₃ ) ₂ I ₂ (0.48 g, 1. 0 mL) was dissolved in 5.0 mL of dimethylacetamide solution, and the mixture was stirred overnight under light shielding. The solution was filtered to remove AgI, and 40 mL of an aqueous solution containing cis-Pt (NH ₃ ) ₂ -IP ₆ (1.2 g, 1.0 mmol) synthesized by the method of Synthesis Example 2-2 described above was added thereto. , Stirred overnight. The filtrate was concentrated under reduced pressure using an evaporator to obtain a yellowish white powder. The precipitated powder was reprecipitated with H ₂ O and methanol to obtain the desired product (yield 1.16 g, yield 77%).

Elemental analysis _{C 7 H 42 N 4 O 34} P 6 Pt 2 Na 8 ((Pt (NH 3) 2) 2 -IP 6 · 8Na · 10H 2 0 · 1MeOH)
Measured value: C 5.60%, H 2.70%, N 3.70%
Calculated values: C 5.59%, H 2.78%, N 3.58%
X-ray fluorescence analysis Measured values: P 6.00, Na 8.54, Pt 1.73
Calculated values: P 6.00, Na 8.00, Pt 2.00

[Example 2-2: Synthesis of (Pt (NH ₃ ) ₂ · Pt (dach))-IP ₆ ]
(Pt (NH ₃ ) ₂ .Pt (dach))-IP ₆ which is a binuclear anion complex of the present invention was synthesized by the scheme shown in the following chemical formula 13.

AgNO ₃ (0.34 g, 2.0 mmol) was dissolved in 40 mL of H ₂ O, and cis-Pt (dach) I ₂ (0.56 g, 1.0 mmol) synthesized by the method of Synthesis Example 2-3 described above was dissolved therein. 5.0 mL of a dimethylacetamide solution in which was dissolved was added, and the mixture was stirred overnight while protected from light. The solution was filtered to remove AgI, and 40 mL of an aqueous solution containing cis-Pt (NH ₃ ) ₂ -IP ₆ (1.2 g, 1.0 mmol) synthesized by the method of Synthesis Example 2-2 described above was added thereto. , Stirred overnight. The filtrate was concentrated with an evaporator to obtain a yellow powder. The precipitated powder was reprecipitated with H ₂ O and methanol to obtain the desired product (yield 1.22 g, yield 83%).

Elemental analysis C ₁₄ H ₄₆ N ₄ O ₃₄ P ₆ Pt ₂ Na ₈ ((Pt (NH ₃ ) ₂ · Pt (dach)) IP ₆ · 8Na · 10H ₂ 0)
Actual value: C 9.42%, H 2.35%, N 3.12%
Calculated values: C 9.29%, H 2.97%, N 3.61%
X-ray fluorescence analysis Measured values: P 6.00, Na 8.54, Pt 1.73
Calculated values: P 6.00, Na 8.00, Pt 2.00

[Example 2-3: Synthesis of Pt (Pt (dach) -IP ₆ ) ₂ ]
Pt (Pt (dach) -IP ₆ ) ₂ , which is a trinuclear anion complex of the present invention, was synthesized according to the scheme shown in the following chemical formula 14.

AgNO ₃ (0.10 g, 0.6 mmol) was dissolved in 5 mL of H ₂ O, and cis-Pt (dach) -IP ₆ (0.41 g, 0.3 mmol) synthesized by the method of Synthesis Example 2-4 described above was dissolved therein. ) And a 7.5 mL aqueous solution of K ₂ PtCl ₄ (0.06 g, 0.15 mmol) were added, and the mixture was stirred in a constant temperature bath at 55 ° C. for 3 nights. The solution was filtered to remove AgCl, the filtrate was concentrated under reduced pressure with an evaporator, and methanol was added thereto to obtain a yellow powder. The precipitated powder was reprecipitated with H ₂ O and methanol to obtain the desired product (yield 337 mg, yield 39%).

Elemental analysis _{C 24 H 94 N 4 Na 14} O 73 P 12 Pt 3
Found: C 9.77, H 2.87, N 1.45
Calculated values: C 9.99, H 3.28, N 1.94

[Example 2-4: Synthesis of 2Pt (dach) ₂ -PP]
2Pt (dach) ₂ -PP, which is a binuclear anion complex of the present invention, was synthesized according to the scheme shown in the following chemical formula 15.

Ag ₂ SO ₄ 31.5 mg of (0.098 mmol) was suspended in _H 2 O 4 mL, this above-synthesized by the method of Synthesis Example 2-3 was the _cis-Pt and (dach) I 2 57mg (0.1mmol ) 1 mL of dissolved dimethylacetamide solution was added, and the mixture was stirred for 4 hours while protected from light. To the solution from which AgI was removed by filtration, 31.5 mg (0.098 mmol) of Ba (OH) ₂ .8H ₂ O was added and stirred for 30 minutes. To the solution from which BaSO ₄ was removed by filtration, 54 mg (0.1 mmol) of cis-Pt (dach) -PP synthesized by the method of Synthesis Example 2-5 described above was added and stirred for 1 day. The filtrate was concentrated with an evaporator to obtain a dark brown powder. The precipitated powder was reprecipitated with H ₂ O and acetone to obtain the desired product (yield 34 mg, yield 40%).

Elemental analysis _{C 12 H 28 N 4 O 7} P 2 Pt 2 · 3H 2 O
Actual value: C 16.71%, H 4.12%, N 6.59%
Calculated values: C 17.01%, H 4.02%, N 6.61%
[Example 2-5: Synthesis of (Pt (dach) · Pt (NH ₃ ) ₂ ) -IP ₆ ]
(Pt (dach) · Pt (NH ₃ ) ₂ ) -IP ₆ which is a binuclear anion complex of the present invention was synthesized by the scheme shown in the following chemical formula 16.

AgNO ₃ (0.17 g, 1.0 mmol) was dissolved in 20 mL of H ₂ O, and cis-Pt (NH ₃ ) ₂ I ₂ (0.24 g, 0.5 mmol) synthesized by the method of Synthesis Example 2-1 described above. 2.5 mL of dimethylacetamide solution was added, and the mixture was stirred overnight while protected from light. The solution was filtered to remove AgI, and 20 mL of an aqueous solution containing cis-Pt (dach) -IP ₆ (0.58 g, 0.5 mmol) synthesized by the method of Synthesis Example 2-4 described above was added and stirred for 3 days. did. The filtrate was concentrated with an evaporator and methanol was added to obtain the desired product as a brown powder (yield 0.32 g, yield 41%).

Elemental analysis _{C 18 H 68 N 4 Na 4} O 39 P 6 Pt 2 ((Pt (dach) · Pt (NH 3) 2) IP 6 · 4Na · 4H · 15H 2 O)
Calculated values: C 10.45%, H 3.64%, N 3.75%
Measured value: C 10.30%, H 3.36%, N 3.37%
[Example 2-6: Synthesis of (Pt (dach)) ₂ -IP ₆ ]
(Pt (dach)) ₂ -IP ₆ which is a binuclear anion complex of the present invention was synthesized according to the scheme shown in the following chemical formula 17.

AgNO ₃ (0.17 g, 1.0 mmol) was dissolved in 20 mL of H ₂ O, and cis-Pt (NH ₃ ) ₂ I ₂ (0.24 g, 0.5 mmol) synthesized by the method of Synthesis Example 2-1 described above. 2.5 mL of dimethylacetamide solution was added, and the mixture was stirred overnight while protected from light. The solution was filtered to remove AgI, and 20 mL of an aqueous solution containing cis-Pt (dach) -IP ₆ (0.58 g, 0.5 mmol) synthesized by the method of Synthesis Example 2-4 described above was added and stirred for 3 days. did. The filtrate was concentrated with an evaporator and methanol was added to obtain the desired product as a brown powder (yield 0.35 g, yield 41%).

Elemental analysis _{C 18 H 64 N 4 Na 8} O 39 P 6 Pt 2 ((Pt (dach) 2) IP 6 · 8Na · 15H 2 O)
Calculated values: C 12.56%, H 3.75%, N 3.26%
Found: C 12.67%, H 3.74%, N 3.10%

[Anti-cancer evaluation using human cultured cancer cell panel]
Some complexes synthesized in the above examples (cation complexes: Examples 1-1, 1-2, 1-3, 1-4, 1-5, 1-7, 1-9, 1-10, 1 -11 and 1-13; anion complexes: Examples 2-1 to 2-4) were tested for anti-cancer properties by using a human cultured cancer cell panel (HCC panel) by the Cancer Research Foundation and Cancer Chemotherapy Center. Evaluated. The anticancer evaluation using the HCC panel is a combination of a drug sensitivity test using a plurality of cell lines and an analysis method using a database program. In this example, specifically, lung cancer 7 system, stomach cancer 6 system, colon cancer 5 system, ovarian cancer 5 system, brain tumor 6 system, breast cancer 5 system, kidney cancer 2 system, prostate cancer 2 system, and A total of 39 melanoma 1 systems were treated as one panel, and an in vitro drug sensitivity test was conducted to obtain a sensitivity pattern (fingerprint) for the drug. The fingerprint thus obtained is a visual representation of the effective concentration deviation of the sample substance for each individual cancer cell, and it is possible to grasp the sensitivity of each cancer cell to the sample at a glance. It is. Then, by comparing this fingerprint with the data in the database, it is possible to confirm the same or not with the action mode of the existing anticancer agent.

  The sensitivity of sample substances to each cancer cell was measured by the following method (Ministry of Education, Culture, Sports, Science and Technology, Cancer Specific Area Research / Integrated Cancer Chemotherapy Information Support Group website (http: // Quoted from gantoku-shien.jfcr.or.jp/panel.html) (the existence of the information was confirmed on the filing date)).

[Method]
Cancer cells are placed in a 96-well plate, the sample solution is added the next day, and after 2 days of culture, cell proliferation is measured by colorimetric determination with sulforhodamine B. An effective concentration deviation of each cancer cell line with respect to the measured average drug effective concentration of the cancer cell line 39 system is calculated and displayed as a fingerprint.

[Evaluation of compounds]
By comparing the fingerprints of the samples with the fingerprints of about 70 kinds of standard anticancer drugs that have been measured and databased, the mechanism of action of the samples is estimated or the novelty of the mechanism of action is evaluated.

FIG. 1 shows the fingerprint for Pt (5-MP) (AtC3) Cl ₂ synthesized in Example 1-5. As shown in FIG. 1, the fingerprint includes three graphs. From the left, these three graphs show GI50 (concentration that suppresses cell growth to 50% compared to control) and TGI (concentration that suppresses proliferation to the same cell number as that at zero (apparent cell number). Concentration with no increase / decrease)), LC50 (concentration that reduces the cell number to 50% of the number of cells at zero (indicating the cell killing effect)) each logarithm (log ₁₀ ) value. The “number of cells at time zero” means the number of cells immediately before administration of the drug sample. In each graph, the vertical axis corresponds to a 39-line cell line, and the horizontal axis represents a value larger than the average value with the average value of each index for the 39-line cell line as the origin. The graph extends to the positive side, and to the negative side when it shows a small value. In addition, Table 1 to Table 2 show IC ₅₀ values obtained using this cancer cell panel. This IC ₅₀ value is a graph in which cell viability (= number of cells after drug treatment / number of cells untreated with drug) at five concentrations from 100 μM to 0.01 μM is plotted against the concentration (common logarithm). From this, it is the value obtained as the drug concentration when the cell viability is 50%.

Here, among the fingerprints for Pt (5-MP) (AtC3) Cl ₂ shown in FIG. 1, a graph of “GI50” is obtained by comparing the known platinum complex anticancer agents cisplatin, carboplatin and oxaliplatin with “ It is shown in FIG. 2 along with the graph of “GI50”. In general, it is presumed that the action modes of anticancer agents whose fingerprints show similar profiles are similar. From this point of view, FIG. 2 shows that the fingerprint profile of Pt (5-MP) (AtC3) Cl ₂ is not similar to any of the profiles of the three known anticancer agents. Therefore, it is suggested that Pt (5-MP) (AtC3) Cl ₂ exhibits anticancer properties by a mode of action different from these known anticancer agents. In addition, all the other cation complexes and anion complexes synthesized above showed a fingerprint profile different from the above-mentioned three known platinum complex anticancer agents.

  This evaluation database by the HCC panel also includes a function called “COMPARE program” for more objectively evaluating the similarity of the mode of action with known anticancer agents. This COMPARE program tests the statistical correlation between two drugs. When a fingerprint of a sample drug is input, a fingerprint similar to that sample drug is collected from the data of standard drugs accumulated in the past. The drugs they have are listed in descending order of similarity. At this time, a correlation coefficient (r value) is calculated for the sample for the purpose of evaluating the similarity. The closer the r value is to 1, the more the sample drug is combined with the existing anticancer agent and the mode of action. It will be similar in this respect. Here, of the complexes synthesized above, those having the largest r value among those listed by the COMPARE program are shown in Tables 1 and 2 below.

[Inhibition assay of cancer-related enzymes]
Some complexes synthesized in the above examples (cation complexes: Examples 1-1, 1-2, 1-3, 1-4, 1-5, 1-7, 1-9, 1-10, 1 -11 and 1-13; anion complexes: Examples 2-1 to 2-4) were examined for inhibitory activity against cancer-related enzymes. Specifically, the inhibitory activity against each of proteasome, telomerase, farnesyltransferase (FPTase), histone deacetylase (HDAC), and protein kinase was evaluated. The results are shown in Tables 1-2 below.

  The inhibitory activity for each enzyme was measured by the following method (Ministry of Education, Culture, Sports, Science and Technology: Cancer Specific Area Research / Integrated Cancer Chemotherapy Information Support Group website (http: // gantoku-shien. Quoted from jfcr.or.jp/panel.html) (the existence of the information was confirmed on the filing date)).

[Proteasome inhibitory activity assay]
[Method]
As a first step, the inhibitory activity of the 20S proteasome on the chymotrypsin-like activity is examined, and the specimen in which a significant inhibitory activity is observed is assayed in the second step. In the second step, caspase-like and trypsin-like activities, as well as inhibitory activity against other proteases (α chymotrypsin, cathepsin B) are measured, and the selectivity of the inhibitory activity of the specimen is evaluated. As a positive control of the inhibitory activity, MG132 of known proteasome inhibitor and clast-Lactacytin β-lactone (Lactacytin activator) are used. In the actual reaction, a sample is added to the 20S proteasome at a concentration of 10 μM, incubated at 30 ° C. for 10 minutes, then added with a fluorescent-labeled peptide containing a 20S proteasome cleavage sequence as a substrate, and reacted at 30 ° C. for 1 hour. At this time, the enzyme activity is measured by quantifying the released fluorescent substance (AMC).

[Evaluation of activity]
Examine the presence or absence of inhibition of each activity based on the untreated control group. For samples that showed significant inhibitory activity, a 50% inhibitory concentration (IC ₅₀ ) was determined by making a serial dilution series (Note: IC ₅₀ values calculated thereby are shown in Tables 1-2. Those that did not show inhibitory activity are indicated by "-" in Tables 1-2).

[Test of telomerase inhibitory activity]
[Method]
An extract of human leukemia U937 cells is used as an enzyme source. This is added to an enzyme reaction solution containing an oligonucleotide serving as a telomerase substrate and a sample compound, and an enzyme reaction is performed at 20 ° C. for 30 minutes. After stopping the reaction on ice, the telomere extension product is amplified by PCR (PCR is carried out under quantitative conditions. A non-telomere DNA fragment is added to the reaction solution in advance, and this is also amplified simultaneously as an internal standard). After the reaction solution is electrophoresed on a polyacrylamide gel, CYBR Green-stained DNA is quantitatively analyzed by densitography. The telomerase inhibitor MST-312 is used as a positive control for inhibitory activity.

[Evaluation of activity]
The sample is added to a final concentration of 10 μM and the inhibitory activity is measured. For samples that showed an inhibitory activity of 50% or more at this concentration and a non-specific inhibitory activity against PCR of less than 40%, a 10-fold dilution series of 10 times from 10 μM was prepared, and the inhibitory activity was measured again (Note) : IC ₅₀ values calculated in this manner are shown in Tables 1 and 2. Those not showing inhibitory activity are indicated by “-” in Tables 1 and 2).

[Farnesyltransferase (FPTase) inhibitory activity assay]
[Method]
FPase: A sample was added to an enzyme reaction solution containing crude purified FPase of human squamous cell carcinoma A431 cells as an enzyme source and GST-H-Ras protein and [ ³ H] -FPP as a substrate, and the mixture was incubated at 37 ° C. for 1 hour. Enzymatic reaction is performed. The reaction is stopped by adding a methanol solution containing 30% TCA and 1% SDS, and left on ice for 1 hour. The acid-insoluble fraction containing [ ³ H] -FPP-bound GST-H-Ras is trapped on a glass fiber filter and washed with 6% TCA. After drying the filter, the radioactivity contained in the acid-insoluble fraction is measured with a liquid scintillation counter to obtain FPase activity. FTI-276 is used as a positive / positive control.

[Evaluation of activity]
In the FPase assay system, the sample is added to a final concentration of 10 μM, and the inhibitory activity is measured. For samples that showed inhibitory activity of 70% or more at this concentration, a 10-fold 10-fold dilution series was prepared from 10 μM, and the inhibitory activity was measured again (Note: IC ₅₀ values calculated thereby are shown in Tables 1-2. In addition, the thing which did not show inhibitory activity is shown by "-" in Tables 1-2).

[Assay for histone deacetylase (HDAC) inhibitory activity]
[Method]
Recombinant human HDAC1 (Class I) is used to test the presence or absence of enzyme inhibitory activity of the sample compound. In the actual reaction, a fluorescent labeled peptide acetylated as a substrate is added to purified human HDAC1 and reacted at 37 ° C. for 30 minutes. Thereafter, trypsin is added, and the enzyme activity is measured by quantifying the fluorescent substance (aminomethylcoumarin) released at this time. Trichostatin A is used as a positive control for inhibitory activity.

[Evaluation of activity]
Samples were first assayed at a final concentration of 100 μM, and serial dilution series were prepared for those in which inhibitory activity was observed, and 50% inhibitory concentrations were determined (Note: IC ₅₀ values calculated thereby are shown in Tables 1-2. In addition, the thing which did not show inhibitory activity is shown by "-" in Tables 1-2).

[Protein kinase inhibition assay (I)]
Here, the inhibitory activity of tyrosine kinases of EGF receptor (experimental system 1) and VEGF receptor (Flt-1) (experimental system 2) is examined by a simple in vitro autophosphorylation reaction.

[Method]
Experimental system 1:
A431 cells are disrupted with a sucrose buffer, and after enucleation, the membrane fraction is collected by ultracentrifugation and used as an enzyme solution. A sample and EGF are added thereto, and the mixture is incubated at 25 ° C. for 30 minutes, and then ATP is added and reacted at 0 ° C. for 15 minutes. After stopping the reaction, the phosphorylated receptor is analyzed by Western blot.
Experimental system 2:
The purified VEGF receptor is used as an enzyme solution. The sample and ATP are added to this and reacted at 30 ° C. for 20 minutes. After stopping the reaction, the phosphorylated receptor is analyzed by Western blot.

[Evaluation of activity]
The sample is added to a final concentration of 10 μM, and the presence or absence of phosphorylation inhibition is determined. For a sample having inhibitory activity, a 10-fold dilution series of 3 or more steps is prepared and tested again, and a sample that is inhibited by 50% or more at a concentration of 1 μM or less is determined to be effective.

[Protein kinase inhibition assay (II)]
Here, cells are stimulated with platelet-derived growth factor (PDGF) to activate PDGF receptor tyrosine kinase and major intracellular signaling pathways (PI3 kinase-AKT pathway, classical MAP kinase pathway, PLC -Evaluate the inhibitory effect of the specimen on (PKC pathway).

[Method]
NRK cells are seeded in a 96-well plate, cultured for 3 days, added with a sample solution, treated for 3 hours, and then stimulated with PDGF. Prepare samples for electrophoresis and perform Western blotting with antibodies against phosphorylated signal transduction molecules (AKT, ERK, PKD, PLCγ1, S6 ribosomal protein) and phosphotyrosine to evaluate the effect on intracellular signal transduction from PDGF receptor To do.

[Evaluation of activity]
The sample is added so as to have a final concentration of 1 and 10 μM, and the presence / absence of inhibition and the inhibition pattern are determined (Note: This inhibition pattern is shown in Tables 1-2. -").

From the IC ₅₀ results shown in Table 1 and Table 2, it can be seen that all of the complexes of the present invention show extremely low values as average values of IC ₅₀ values for 39-series cancer cell lines. Here, the IC ₅₀ value calculated by the same method for cisplatin, which is a commonly used platinum preparation, was 8 μM. Since the IC ₅₀ value of many complexes of the present invention is significantly lower than the IC ₅₀ value of cisplatin, the complex of the present invention is expected to be used as a highly active novel anticancer agent.

  Moreover, from the result of the r value (maximum value) shown in Table 1 and Table 2, in the complex of the present invention, the r value is at most about 0.7, and from the viewpoint of the mode of action with existing anticancer agents. It can be seen that no similarity was observed. From this, it is considered that the complex of the present invention exhibits an anticancer action based on a different mode of action from the existing anticancer agents. Therefore, the complex of the present invention is expected as an effective anticancer agent for so-called “drug resistant cancer” in which existing anticancer agents are not effective. In particular, in the cation complex of the present invention, the r value is less than 0.5 in almost half of the examples, which is very promising as a drug-resistant cancer therapeutic agent.

  Furthermore, as shown in Tables 1 and 2, all of the complexes of the present invention showed an inhibitory effect on proteasomes. The proteasome is an enzyme that selectively degrades polyubiquitinated intracellular proteins, and plays a central role in the degradation of proteins that control the cell cycle and apoptosis. For this reason, it is considered that this proteasome inhibitory action is involved in at least part of the mechanism by which the complex of the present invention exhibits anticancer properties. In addition, most of the complexes of the present invention inhibit one or more of various cancer-related enzymes such as telomerase, farnesyltransferase, histone deacetylase, and protein kinase. From this, it is considered that the anticancer property of the complex of the present invention is exhibited as a comprehensive action in combination with not only the above-described inhibitory action of the proteasome but also the inhibitory action against other enzymes. .

[Evaluation of nephrotoxicity]
The nephrotoxicity of several complexes synthesized in the above examples was evaluated by the following method. In addition, cisplatin was used as a control.

Five 6-week-old ICR male mice were taken as a group and blood was collected at 0 hours before administration and 72 hours after drug intravenous administration, and the values of creatinine and BUN, which are indicators of renal function, were examined. The sample was fasted 16 hours before blood collection, and iStat was used to measure creatinine and BUN. In addition, the administered drug amount was the same mole for each drug. The types of drugs evaluated and their dosages are as follows:
Pt (5-MP) (AtC3) (Example 1-5): 28.5 mg / kg
(Pt (NH ₃ ) ₂ · Pt (dach))-IP ₆ (Example 2-2): 53 mg / kg
_{Pt (Pt (dach) -IP 6} ) 2 ( Example 2-3): 94mg / kg
Cisplatin: 10 mg / kg.

Pt (5-MP) (AtC3) (Example 1-5), (Pt (NH ₃ ) ₂ .Pt (dach))-IP ₆ (Example 2-2), Pt (Pt (dach) -IP _{6 2} (Example 2-3) and the results of nephrotoxicity evaluation of cisplatin are shown in FIGS.

  As shown in FIG. 6, 72 hours after cisplatin administration, the BUN value increased approximately twice and the creatinine value increased approximately 3.5 times compared to before administration, and the renal function was impaired. I understand that. On the other hand, as shown in FIGS. 3 to 5, in the case of the complex of the present invention, no significant increase in BUN value or creatinine value was observed even 72 hours after administration.

Similarly, nephrotoxicity of the complex of the present invention was evaluated using 6-week-old male ICR mice (n = 3 to 5 for each group). Specifically, first, the mouse was fasted, and after 16 hours, blood was collected from the vein located behind the tibia using an animal lancet, and the components in the blood (sodium Na, The values of potassium K, ionized calcium iCa, total carbon dioxide tCO ₂ , glucose Glc, blood urea nitrogen BUN, creatinine Crea, hematocrit Hct, hemoglobin Hb) were measured. Thereafter, cisplatin 33 μmol / kg and Pt (Pt (dach) -IP ₆ ) ₂ 33 μmol / kg were administered from the tail vein. After 56 hours, fasting was started, and further 16 hours later, blood was collected in the same manner, and blood components were measured.

FIG. 7 is a graph showing the mean ± standard deviation (relative value) of blood glucose values measured for each sample administration group. As shown in FIG. 7, the blood glucose level (Glc) decreased in the control. This is thought to be due to stress. In addition, in Pt (Pt (dach) -IP ₆ ) ₂ and (Pt (dach) · Pt (NH ₃ ) ₂ ) -IP ₆ , no decrease in Glc was observed, and a significant difference of 1% from the control was observed. On the other hand, (Pt (dach) · Pt (NH ₃ ) ₂ ) -IP ₆ and (Pt (dach)) ₂ -IP ₆ showed a decrease in Glc, and no significant difference from the control was observed.

FIG. 8 is a graph showing the mean ± standard deviation (relative value) of blood BUN values measured for each sample administration group. FIG. 9 is a graph showing the mean ± standard deviation (relative value) of blood Crea values measured for each sample administration group. As shown in FIGS. 8 and 9, cisplatin showed an increase in BUN and Crea, which are renal function markers, but such an increase in BUN and Crea was not observed in the complex of the present invention. This is considered to be due to the fact that the complex of the present invention accumulates in the bone and the accumulation in the kidney is reduced, and that the water excretion is high and renal excretion is promoted. In the measurement results of BUN, there was a decreasing trend for Pt (Pt (dach) -IP ₆ ) ₂ and (Pt (dach)) ₂ -IP ₆ . Since these complexes have particularly strong cell growth-inhibiting effects, it is speculated that loss of appetite, which is one of the side effects of platinum anticancer drugs, is stronger than other complexes, resulting in a decrease in BUN.

FIG. 10 is a graph showing the mean ± standard deviation (relative value) of blood hematocrit (Hct) values measured for each sample administration group. FIG. 11 is a graph showing the mean ± standard deviation (relative value) of blood hemoglobin (Hb) values measured for each sample administration group. As shown in FIGS. 10 and 11, no significant difference was observed between the control and the complex in the measurement results of Hct and Hb. However, Hct showed a decreasing trend except for Pt (Pt (dach) -IP ₆ ) ₂ . The complex of the present invention is expected to accumulate in bone and is expected to affect bone marrow.

As described above, in the measured parameters, except for Glc, Pt (Pt (dach) -IP ₆ ) ₂ , (Pt (NH ₃ ) ₂ · Pt (dach))-IP ₆ , (Pt (dach) · Pt ( There was no significant difference between NH ₃ ) ₂ ) -IP ₆ and (Pt (dach)) ₂ -IP ₆ . These complexes of the present invention have no significant difference in in vitro cell growth-inhibiting activity or adsorption ability to hydroxyapatite (HAP), and it is considered that there was no difference in the effect on the living body.

  From the above, the complex of the present invention has a very low risk of impairing renal function despite the fact that both the cation complex and the anion complex contain noble metals such as platinum. Therefore, it can be a promising candidate for an anticancer drug with extremely reduced side effects.

[Evaluation of adsorptivity to hydroxyapatite (HAP)]
The adsorptivity to hydroxyapatite (HAP; Ca ₁₀ (PO ₄ ) ₆ (OH) ₂ ) was evaluated for the four types of anion complexes synthesized in Examples 2-1 to 2-2 and 2-5. HAP is a main component constituting human teeth and bones.

  This evaluation was performed according to the Langmuir theory which is a monomolecular layer adsorption theory. First, the Langmuir theory will be briefly described.

  When monolayer adsorption occurs between the adsorbed substance and the adsorbent, there is a linear relationship between the concentration of the compound remaining in the solution and the amount of adsorption, and monolayer equilibrium adsorption. Indicates that the reaction is in progress. It is known that when the monomolecular layer adsorption occurs, the Langmuir monomolecular layer adsorption formula of the following formula (1) is established.

  In Equation (1), W represents the mass of the solute adsorbed per unit weight of the adsorbent, Ws represents the saturated adsorption amount, a represents the adsorption equilibrium constant, and C represents the equilibrium concentration of the solute. Here, when the formula (1) is modified, the following formula (2) is derived.

  Based on this equation (2), the slope of the Langmuir monolayer adsorption equation (1 / aWs) and the y-intercept (1 / C plotted on the horizontal axis and 1 / W plotted on the vertical axis) From each value of 1 / Ws), the adsorption equilibrium constant a and the saturated adsorption amount Ws in the monolayer adsorption state are calculated.

_{[(Pt (NH 3) 2} ) Evaluation of HAP adsorptive 2 -IP _6]
According to the above-described Langmuir theory, the adsorptivity of (Pt (NH ₃ ) ₂ ) ₂ -IP ₆ synthesized in Example 2-1 to HAP was evaluated. The specific method is as follows.

To 5 mg of HAP, 5 mM HEPES buffer (pH 7.5) was added and incubated at 37 ° C. for 30 minutes. Samples were prepared by adding a solution of (Pt (NH ₃ ) ₂ ) ₂ -IP _{6 in} the same buffer solution to a total concentration of 5.0 mL at a complex concentration of 1, 2, 3 or 4 mM. did. These were incubated for 1.5 hours and separated into HAP and solution by centrifugation. Next, ³¹ P-NMR of the recovered HAP was measured, and the amount of (Pt (NH ₃ ) ₂ ) ₂ -IP ₆ adsorbed on HAP at each complex concentration was determined. Based on the obtained results, the concentration C (mM) of the compound remaining in the solution and the number of moles W (μmol / mg) of the compound adsorbed per 1 mg of HAP were calculated, respectively. The data was plotted according to 2) to create a graph. The adsorption equilibrium constant a and the saturated adsorption amount Ws ₂ were calculated from the slope of the straight line and the value of the y-intercept in this graph. The results obtained are shown in Table 3 below. Table 3 also shows the results of a similar evaluation of cisplatin and carboplatin, which are known platinum complex anticancer agents.

From the results shown in Table 3, it can be seen that all of the anion complexes of the present invention exhibit a larger adsorption equilibrium constant a as compared with cisplatin and carboplatin. This indicates that the anion complex of the present invention has a very high affinity for HAP. As described above, since HAP is a major component of human teeth and bones, the anion complex of the present invention having an IP ₆ structure alone or complexed with HAP It can be used as a drug delivery system (DDS) formulation that can be targeted to cells affected by bone-related diseases such as cancer.

[Evaluation of in vivo anticancer effect (cationic complex)]
In order to examine the in vivo anticancer effect of the cation complex of the present invention, nude mice were synthesized in Example 1-5 as samples using BALB / c Slc-nu / nu as detailed below. In vivo anticancer activity of Pt (5-MP) (AtC3) Cl ₂ was evaluated.

Passage experiment of human prostate cancer-derived cell DU-145 In order to evaluate the anti-cancer activity of cation complexes in vivo by creating a gall cancer mouse using human prostate cancer-derived cell DU-145, A culture experiment of DU-145 was conducted. The operation procedure of the passage experiment is shown below.

Preparation of CO ₂ incubator
A CO ₂ cylinder was connected to the incubator and adjusted to 5%, 37 ° C. Inside the incubator was installed a vat containing 1000 ml of sterilized distilled water plus disinfectant hibiten.

Preparation of a clean bench The sterilization lamp of the clean bench was turned off, 70% ethanol was sprayed onto the hand, the inside of the bench was wiped with a Kim wipe soaked in ethanol, and sterilized.

Medium creation
10.4 g of RPMI-1640 medium was added to 820 ml of MilliQ water and stirred. Thereto, a penicillin-streptomycin mixed solution was added so as to be penicillin 100 units / μl and streptomycin 100 mg / μl. Further, 2.0 g of NaHCO ₃ and 100 ml of FBS were added and stirred, and then sterilized distilled water was added so that the total amount became 1000 ml. Thereafter, it was sterilized and filtered in a clean bench and used as a medium for the following culture experiments.

Preparation of PBS 9.6 g of Dulbecco's Phosphate Buffered Saline was added to 1000 ml of sterilized distilled water, and the whole was dissolved and then sterilized at 120 ° C. for 20 minutes.

Cell Culture and Passage Human prostate cancer-derived cells DU-145 that had been cryopreserved in sample containers were rapidly lysed at 37 ° C. and transferred to a dish containing 25 ml of medium in a clean bench. Two days later, the medium was removed, washed with 10 ml of PBS, 5 ml of a 5-fold diluted 0.5% -Trypsin / 5.3 mM-EDTA solution was added, and the mixture was incubated at 37 ° C. for 10 minutes. After confirming that all cells were detached, 5 ml of medium was added and subcultured. Two days later, the same operation was performed.

Measurement of the number of cells A cell suspension stained with trypan blue was placed on a counting plate, and a cover glass was applied from above. The number of cells present in a total of eight counter parts of the counting plate was counted with an optical microscope, the average number of cells present in one place was calculated, and the number of cells in the cell suspension before dilution was calculated.

Preparation and injection of cell suspension A cell suspension was prepared so that the number of cells was 8.3 × 10 ⁶ cells per 100 μl of medium. 100 μl of the prepared cell suspension was injected subcutaneously near the right shoulder of the mouse.

Sample administration and measurement of tumor size 20 days after the tumor was injected into the mouse, the experiment was started on day 0, and distilled water (control) or Pt (5-MP) (AtC3) Cl ₂ was used on days 0, 7, and 14. Was administered. Each sample (n = 5) was administered at 8.25 mmol / kg. Measure body weight (mouse volume) and tumor size on day 0, day 1, day 3, day 7, day 8, day 11, day 14, day 15, day 18, day 20 did. Tumor size was measured using calipers, and the mouse volume before drug administration of the tumor volume obtained by A x B ² x 0.52 (A = tumor length, B = tumor width; all units are mm) Tumor volume ratio was calculated as a percentage of.

FIG. 12 is a graph showing changes in the mean tumor volume ratio of bile cancer mice administered with Pt (5-MP) (AtC3) Cl ₂ in comparison with the control group. As shown in FIG. 12, when Pt (5-MP) (AtC3) Cl ₂ was administered once a week at a dose of 8.25 mmol / kg, there was a tendency to suppress the growth of tumor cells compared to control (distilled water). It was seen. However, no significant difference was observed even 20 days after the start of the experiment. In addition, at each timing when the tumor volume ratio of the mouse was calculated as described above, the weight ratio value of the bile cancer mouse (relative value of the mouse weight when the weight on the 0th day is 1) was also calculated. FIG. 13 is a graph showing the change in the average body weight ratio of the bile cancer mice administered with Pt (5-MP) (AtC3) Cl ₂ in comparison with the control group. As shown in FIG. 13, since there was no difference in the average value of the weight ratios of the mice, the side effects due to the administration of Pt (5-MP) (AtC3) Cl ₂ were shown to be small.

Subsequently, an experiment similar to the above was performed using distilled water (control), cisplatin, and Pd (5-MP) (AtC3) (NO ₃ ) ₂ as samples.

FIG. 14 is a graph showing changes in the mean tumor volume ratio of bile cancer mice administered with cisplatin or Pd (5-MP) (AtC3) (NO ₃ ) ₂ compared to the control group. As shown in FIG. 14, when Pd (5-MP) (AtC3) (NO ₃ ) ₂ was administered once a week at a dose of 8.25 mmol / kg, the growth of tumor cells was suppressed compared to control (distilled water). The degree was similar to that of cisplatin. However, no significant difference was observed even 20 days after the start of the experiment. FIG. 15 is a graph showing the change in the average body weight ratio of the bile cancer mice administered with cisplatin or Pd (5-MP) (AtC3) (NO ₃ ) ₂ compared with the control group. As shown in FIG. 15, since there was no difference in the average value of the body weight ratio of the mice, the administration of Pd (5-MP) (AtC3) (NO ₃ ) _{2 as} a Pd complex was the same as that of the Pt complex. It was shown that side effects were small.

[Evaluation of in vivo anticancer effect (anion complex)]
In order to examine the in vivo anticancer effect of the anion complex of the present invention, the in vivo anticancer activity of Pt (Pt (dach) -IP ₆ ) ₂ synthesized in Example 2-3 as a sample was evaluated. did. In addition, although the experimental protocol of an anion complex is as follows, about the matter which is not described, the method similar to having mentioned above about the cation complex of this invention was employ | adopted.

Day 0 mouse delivery: BALB / c slc-nu / nu male 4-week-old cell planting: DU-145 3.8 × 10 ⁶ cells / mouse subcutaneously on right shoulder
Day 24 Drug administration (intravenous administration)
Cisplatin 17 μmol / kg administration group n = 6
Control (distilled water administration) group n = 5
Pt (Pt (dach) -IP ₆ ) ₂ 17 μmol / kg administration group n = 4
Day 42 Drug administration (intravenous administration)
Pt (Pt (dach) -IP ₆ ) ₂ 17 μmol / kg re-administered to the 17 μmol / kg group (not re-administered to the cisplatin group and control group)
FIG. 16 is a graph showing changes in the average tumor volume ratio of bile cancer mice administered with cisplatin or Pt (Pt (dach) -IP ₆ ) ₂ in comparison with the control group. As shown in FIG. 16, the cisplatin 17 μmol / kg once-administered group is not so different from the control, whereas the Pt (Pt (dach) -IP ₆ ) ₂ 17 μmol / kg twice-administered group is DU-145. There was a tendency to suppress the growth of prostate cancer. In the cisplatin-administered group, the initial weight loss was large, so the second dose was not administered in order to avoid death due to the second dose.

[In vivo evaluation using rat bone metastasis model]
Confirmation of bone metastasis model rat preparation is that the bone surface becomes rough after n-1 days (n-1 = 17, 21) compared to the right foot tibia without transplanted cancer cells and the cancer cells Was confirmed by the proliferation of bone metabolism.

On day n (n = 18, 22) after transplantation of cancer cells, Pt (Pt (dach) -IP ₆ ) ₂ or cisplatin was administered, and tumor size, response to foot stimulation, body weight before and after administration From the changes, tumor growth inhibitory effect, pain alleviation effect, and side effects were evaluated. As described later, various evaluation methods were performed using calipers for tumor size, and reactivity to foot stimulation was performed using von frey filament.

Preparation of medium solution RPMI-1640 medium 5.2 mg, NaHCO ₃ 1.0 mg, Fetal Bovine serum (FBS) 50 ml dissolved in approximately 450 ml MilliQ water, penicillin-streptomycin mixed solution (penicillin 10000 units / ml, 5 ml of streptomycin (10000 μg / ml) was added and stirred. Thereafter, it was sterilized and filtered in a clean bench.

Preparation of PBS solution
9.6 g of Phosphate Buffered Saline (PBS) was added to 1 L of MiliQ water, stirred and then autoclaved.

MRMT-1 cell culture The cryopreserved MRMT-1 cells were thawed in a 37 ° C constant temperature bath. Thereafter, 20 ml of the prepared medium was added to the dish in a clean bench, and pipetted well there, and 1 ml of cells was added. After that, it was stored in a CO ₂ incubator. Two days later, a passage experiment was performed. The stored dish was washed with PBS in a clean bench, added with 10 ml of trypsin, placed in a CO ₂ incubator for about 20 minutes, and the cells were detached from the dish. Next, half of the detached cells were discarded in a clean bench, 20 ml of medium was added to the remaining dish, and then the dish was stored in a CO ₂ incubator. This passage experiment was performed three times.

Creation of bone metastasis model
Under a Nembutal anesthesia, a 7-week-old female SD rat was incised so that the proximal part of the tibia was visible, and a hole reaching the bone marrow cavity was drilled using a 23-gauge needle 5 mm distal to the left knee joint. MRMT-1 rat breast cancer cells (3000 cells / 3 μL) were injected using a Hamilton syringe. On the other hand, a hole was made in the corresponding part of the right foot tibia, and only 3 μL of the same volume of medium was administered as a pseudo treatment. After the operation, the bone hole was filled with bone wax (beeswax), and the wound was sutured.

Confirmation of bone metastasis model preparation After surgery, rats were radiographed over time. After anesthetizing the rats with Nembutal, the rats were affixed to a cassette with an IP (imaging plate) inserted, and X-rays were taken with SOFTEX M-6, and then IP was analyzed with BAS5000.

・ Evaluation of tumor growth inhibition effect, pain relief, and side effects Pt (Pt (dach) -IP ₆ ) ₂ m mol / kg, cisplatin m mol / n n days after transplantation of cancer cells into rat tibia (n = 18, 22) A solution of kg (m = 33, 8.25) dissolved in 50 µl of water was administered from the tail vein. In addition, 100 μl of water was administered to the control group.

  In order to evaluate the pain relieving effect, von frey filament was used. The von frey filament test is a typical method for measuring mechanical allodynia. The sole of the foot of the foot was stimulated with the filament, pressure was gradually applied, and the pressure at which the rat showed a painful reaction was taken as a threshold value. This operation was measured for each rat, 5 times for each left foot and right foot, with 5 minutes alternately (left foot measurement → 5 minutes → right foot measurement → 5 minutes →) × 5 times / 1 rat). The left / right foot ratio (right foot / left foot) of the median value was calculated, and this value was used as an index of pain. The ratio of the values was evaluated relative to the value of the right foot / left foot before administration as 1.

Tumor size was measured using calipers, tumor volume was calculated by A × B ² × 0.52 (A = tumor length, B = tumor width; units are all mm), and the value one day before sample administration was 1 Evaluation was made using relative values.

FIG. 17 is a graph showing changes in the mean tumor volume ratio of the cisplatin 33 μmol / kg once-administered group and the Pt (Pt (dach) -IP ₆ ) ₂ 33 μmol / kg once-administered group compared with the control group. As shown in FIG. 17, the cisplatin 33 μmol / kg once-administered group was not different from the control, whereas the Pt (Pt (dach) -IP ₆ ) ₂ 33 μmol / kg once-administered group was MRMT-1 breast cancer. There was a tendency to suppress the growth of

FIG. 18 shows the tumor growth inhibitory effect with respect to the cisplatin 8.25 μmol / kg one-time administration group (day 1 administration) and Pt (Pt (dach) -IP ₆ ) ₂ 8.25 μmol / kg two-time administration group (1, 8 days). It is a graph which shows the change of the average tumor volume ratio of eye administration compared with a control group. In this experiment, the day before drug administration was defined as day 0 (the same applies hereinafter).

FIG. 19 shows cisplatin 8.25 μmol / kg once-administered group (administration on the first day) and Pt (Pt (dach) -IP ₆ ) ₂ 8.25 μmol / kg twice-administered group (on days 1 and 8) It is a graph which shows the change of the right and left foot ratio (average value) of administration) compared with a control group.

FIG. 20 shows cisplatin 8.25 μmol / kg administered once (day 1 administration) and Pt (Pt (dach) -IP ₆ ) ₂ 8.25 μmol / kg twice administered group (administered on days 1 and 8). ) Is a graph showing changes in body weight (average value) in comparison with a control group.

From the results shown in FIG. 18 (evaluation of tumor growth inhibitory effect), the cisplatin 8.25 μmol / kg once-administered group appeared to be inferior to the control group, but these differences were within the error range. On the other hand, a tendency to suppress the growth of MRMT-1 breast cancer was observed in the Pt (Pt (dach) -IP ₆ ) ₂ 8.25 μmol / kg twice-administered group. Similar to the evaluation using nude mice, since the initial weight loss was large in the cisplatin administration group, the second administration was not performed for the purpose of avoiding death due to the second administration. On the other hand, Pt (Pt (dach) -IP ₆ ) ₂ had almost the same body weight ratio as the control even when administered twice (FIG. 20). Therefore, the side effect of Pt (Pt (dach) -IP ₆ ) ₂ Was suggested to be small.

In addition, with respect to the results shown in FIG. 19 (pain alleviation effect), the left foot of bone cancer generally becomes more sensitive to pain as the cancer progresses, and the threshold value becomes smaller. As the graph grows, the graph rises to the right. As shown in FIG. 19, the cisplatin-administered group shows a tendency similar to that of the control that rises to the right, whereas Pt (Pt (dach) -IP ₆ ) ₂ shows a substantially flat inclination and tends to relieve pain. Admitted.

Claims (15)

A metal complex represented by the following chemical formula 1 or a salt thereof:
Where
M is Pt or Pd;
R _{1 to} R ₁₇ are each independently a hydrogen atom, an alkyl group having 1 to 30 carbon atoms, a cycloalkyl group having 3 to 30 carbon atoms, an alkenyl group having 2 to 30 carbon atoms, or a cyclohexane having 3 to 30 carbon atoms. Alkenyl group, C2-C30 alkynyl group, C7-C30 aralkyl group, C7-C30 aralkenyl group, C7-C30 aralkynyl group, C6-C30 aryl group, halogen atom Haloalkyl group having 1 to 30 carbon atoms, haloalkenyl group having 2 to 30 carbon atoms, haloalkynyl group having 2 to 30 carbon atoms, haloaryl group having 6 to 30 carbon atoms, alkoxy group having 1 to 30 carbon atoms, carbon number A 6-30 aryloxy group, a hydroxy group, an amino group, a C1-C30 alkylamino group, a C6-C30 arylamino group, a cyano group, or a nitro group. R _{1 to} R ₃ and R _{5 to} R ₁₇ are hydrogen atoms, and R ₄ is an alkyl group having 1 to 30 carbon atoms, a halogen atom, an alkoxy group having 1 to 30 carbon atoms, or a nitro group. 2. The metal complex or salt thereof according to 1.   The metal complex or a salt thereof according to claim 1, wherein an even number of the same alkyl group having 1 to 30 carbon atoms is bonded to a symmetrical position in the phenanthroline ring in Chemical Formula 1. The metal complex or a salt thereof according to claim 2, represented by any one of the following chemical formulas 1a to 1d:
The metal complex or a salt thereof according to claim 3, which is represented by any one of the following chemical formulas 1e to 1g:
  The metal complex or a salt thereof according to any one of claims 1 to 5, wherein M is Pt. A metal complex represented by the following chemical formula 2 or a salt thereof:
Where
M is Pt or Pd;
X is a structure represented by the following chemical formula 3:
Or the structure represented by following Chemical formula 4 derived from myo-inositol 6-phosphate ester:
Or a structure represented by the following chemical formula 5:
Where
M is the same definition as above,
A is a structure represented by the following chemical formula 6 derived from myo-inositol 6-phosphate ester:
Each R is independently a hydrogen atom, or together with the adjacent R, forms a structure represented by the following chemical formula 7:
The metal complex or a salt thereof according to claim 7, which is represented by any one of the following chemical formulas 2a to 2f:
  The metal complex according to claim 7 or 8, wherein X in Chemical Formula 2 is a structure represented by Chemical Formula 4 or Chemical Formula 5 and is combined with hydroxyapatite.   The metal complex or a salt thereof according to any one of claims 7 to 9, wherein M is Pt.   The anticancer agent which contains the metal complex of any one of Claims 1-10, or its salt as an active ingredient.   The proteasome inhibitor which contains the metal complex of any one of Claims 1-10, or its salt as an active ingredient.   One or two selected from the group consisting of telomerase, farnesyltransferase, histone deacetylase, and protein kinase, containing the metal complex according to any one of claims 1 to 10 or a salt thereof as an active ingredient. An enzyme inhibitor that inhibits the above enzymes. A metal complex represented by the following chemical formula 8:
Where
M is Pt or Pd;
Each R is independently a hydrogen atom, or together with the adjacent R, forms a structure represented by the following chemical formula 7:
A metal complex represented by the following chemical formula 2 or a salt thereof by reacting with a halogenated platinum salt or a halogenated palladium salt:
Where
M and R have the same definition as above,
X is a structure represented by the following chemical formula 5:
Where
M is the same definition as above,
A is a structure represented by the following chemical formula 6 derived from myo-inositol 6-phosphate ester:
The manufacturing method of the metal complex or its salt including the process of obtaining.   The process according to claim 14, wherein the reaction is carried out in the presence of a halogen scavenger.