JP6154231B2 - Anticancer drug - Google Patents

Description

This invention relates to an anti-cancer agent.

  Cisplatin (cis-dichlorodiammine platinum (II)), carboplatin (cis-1,1-cyclobutanedicarboxylate diammine platinum (II), see Patent Document 1), nedaplatin (cis-O, O′-glycolate diammine) Platinum preparations such as platinum (II) (see Patent Document 2) have strong anticancer activity, and are currently used as major anticancer agents against many cancers such as lung cancer and malignant lymphoma. These platinum preparations are known to kill cancer cells by strongly binding to the base portion constituting DNA and preventing DNA replication.

  However, conventional platinum preparations have the following problems. First, since platinum is used as a raw material, there has been a problem that the price of the drug becomes high. In addition, some cancer cells have a platinum preparation-resistant cancer that is excreted out of the cell without binding to DNA even when a platinum preparation is administered.

  On the other hand, the inventors have studied a heterocyclic compound having a specific structure, a metal complex having this as a ligand, and the use of the metal complex as an oxidation catalyst, and have already filed patent applications (Patent Documents). 3).

JP 49-48621 JP 59-222497 International application number PCT / JP2013 / 062176

  Thus, an object of the present invention is to provide an anticancer agent that is inexpensive without using expensive noble metals such as platinum and has a different mechanism of action from conventional platinum preparations.

  The inventors have speculated that the metal complex and its analogs can be stably present in water and can be used as an anticancer agent, and as a result of earnest research, the metal complex is found in cancer cells. The present inventors have found that it has excellent cytotoxicity, and have completed the present invention.

  That is, the anticancer agent of the present invention is an anticancer agent containing a metal complex represented by the following general formula (1) as an active ingredient.

(In the formula , M represents one of iron, manganese, and cobalt. X represents a counter ion, and n represents 1 or 2.)

Among them, M is manganese, X is ClO ₄ ^- in which is preferable.

  The anticancer agent of the present invention can be produced at low cost without using platinum, which is a noble metal, as a raw material. In addition, the coordination of the metal complex which is an active ingredient of the anticancer agent of the present invention is generally substitution activity and does not bind to the base constituting DNA. Therefore, cancer cells can be killed by an action mechanism different from that of the platinum preparation, which is also effective for cancer in which the platinum preparation does not exert its effect.

FIG. 1 is a diagram showing a synthesis route of H-dpaq ^{H, ester} , which is a heterocyclic compound constituting the anticancer agent of the present invention.

The present invention, the heterocyclic compounds for anticancer agents as an active ingredient a metal complex having a ligand. Therefore, these will be described in detail below. In addition, this invention is not limited in any meaning by the following description, In the range which does not impair the meaning of this invention except the following illustrations, it can change suitably.

1. Heterocyclic Compound (1) Heterocyclic Compound The heterocyclic compound constituting the anticancer agent of the present invention is a heterocyclic compound represented by the following general formula ( 2 ). This heterocyclic compound is abbreviated as “H-dpaq ^{H, ester} ”. In addition, a compound in which —OCH ₂ CO ₂ C ₂ H _{5 of} “H-dpaq ^{H, ester} ” is replaced with a hydrogen atom is replaced with “H-dpaq ^{H, H} ” or “H-dpaq ^{H, H} ” A compound in which the 8-position is substituted with a nitro group is abbreviated as “H-dpaq ^{NO2, H} ”.

(2) Method for producing heterocyclic compound A method for producing “H-dpaq ^{H, ester} ” ^, which is a heterocyclic compound constituting the anticancer agent of the present invention, will be described below. In addition, the said heterocyclic compound is not limited to what is obtained with the following manufacturing method.

The heterocyclic compound constituting the anticancer agent of the present invention uses an etherified product of 6-hydroxy-8-aminoquinoline represented by the following general formula ( 3 ) as a starting material.

Incidentally, etherification products of the 6-hydroxy-8-aminoquinoline are readily obtained by example as shown in Chemical Formula 4 and Chemical Formula 5 below. Specifically, first, a 4-amino-3-nitrophenol, by Debuna von-Miller (DOEBNER-von MILLER) quinoline Synthesis and Sukuraupu (SKRAUP) known quinoline synthesis of quinoline synthesis 6- hydroxy - 8 - After synthesizing Nitorokinori down, etherifying the 6-position hydroxyl group in the halide of ethyl acetate. Next, as shown in the following chemical formula 5 , the nitro group at the 8-position is reduced to an amino group.

Using the compound represented by the general formula ( 3 ) thus obtained, for example, the following reactions ( 6 ) and ( 7 ) are performed.

Here, X ¹ and X ² are halogen atoms, and preferably both are bromine atoms. Reaction ( 6 ) is an amidation reaction of an amino group, and reaction ( 7 ) is an N-alkylation reaction of an amine with an alkyl halide.

Here, amine used in the reaction (7) is 2,2' Jipikoriruami down.

Regarding heterocyclic compounds other than the above-mentioned “H-dpaq ^{H, ester} ”, for example, “H-dpaq ^{H, H} ”, “H-dpaq ^{NO2, H} ”, “H-dpaq ^{OMe, H} ”, Patent Document 3 And may be synthesized according to the description.

2. Metal complex (1) Metal complex The metal complex which is an active ingredient of the anticancer agent of this invention is represented by the following general formula ( 1 ).

Here, M represents any one of iron, manganese, and cobalt, and manganese is preferable because of its high activity. X represents a counter ion. Specifically, ClO ₄ ⁻ , Cl ⁻ , Br ⁻ , AcO ⁻ , TfO ⁻ , CF ₃ CO ₂ ⁻ , BF ₄ ⁻ , ReO ₄ ⁻ , AsF ₆ ⁻ , SbF ₆ ⁻ Among them, ClO ₄ ⁻ is preferable because of its high activity. Note that n = 1 or 2 .

(2) Method for producing metal complex The metal complex which is an active ingredient of the anticancer agent of the present invention is obtained by coordinating the heterocyclic compound of the present invention to a metal, and the method is particularly limited as long as it is a known method. Can be used without

For example, in a solvent, it is sufficient under the conditions capable of forming a complex coexist and said heterocyclic compound and a predetermined metal ions, specifically, the heterocyclic compound is dissolved in a solvent together with a basic compound, If a predetermined metal ion solution is added, complex microcrystals can be formed.

In addition, as said solvent, polar organic solvents, such as methanol and acetonitrile, are suitable. Further, as the basic compound, triethylamine, N, N-diisopropylethylamine and the like are preferable. Furthermore, after complexed, high purity of the metal complex is obtained by the washing with a solvent such as methanol.

3. Anti-cancer agent The anti-cancer agent of the present invention contains a metal complex represented by the following general formula (1) as an active ingredient, and is used in combination with a metal complex alone or a known pharmaceutical carrier, or a human suffering from cancer or other It is to be administered to animals. The structure, application case, administration mode, etc. will be described in detail below.

Here, M, X, for n will be omitted because it is same as the metal complex.

(1) Application case The kind of cancer to which the anticancer agent of this invention is applied is not particularly limited. Applicable cancer types include malignant melanoma, malignant lymphoma, digestive organ cancer, lung cancer, esophageal cancer, stomach cancer, colon cancer, rectal cancer, colon cancer, ureteral tumor, gallbladder cancer, bile duct cancer, biliary tract cancer, breast cancer, Liver cancer, pancreatic cancer, testicular 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, leukemia, true Examples include polycythemia, neuroblastoma, retinoblastoma, myeloma, cystoma, sarcoma, osteosarcoma, myoma, skin cancer, basal cell cancer, skin appendage cancer, skin metastasis cancer, skin melanoma, etc. It can be applied not only to malignant tumors but also to benign tumors. Further, the anticancer agent of the present invention can be used for suppressing cancer metastasis, and can also be used as a postoperative cancer metastasis inhibitor.

(2) Administration form etc. The anticancer agent of this invention can be administered to humans or animals in various forms, and the administration form is not particularly limited. Specifically, it may be oral administration, or parenteral administration such as intravenous, intramuscular, subcutaneous or intradermal injection, rectal administration or transmucosal administration.

  Examples of the dosage form suitable for oral administration include tablets, pills, granules, powders, capsules, solutions, suspensions, emulsions, syrups and the like, and pharmaceutical compositions suitable for parenteral administration include injections. And transdermal agents such as drops, nasal drops, sprays, inhalants, suppositories, ointments, creams, powdery coatings, liquid coatings, patches and the like. Furthermore, it may be a sustained-release preparation by embedding pellets or a known technique. In addition, a preferable dosage form, formulation form, etc. may be appropriately selected by a doctor according to the age, sex, constitution, symptom, treatment timing, etc. of the patient.

  When the anticancer agent of the present invention is used as a solid preparation such as a tablet, pill, powder, powder, granule or the like, the metal complex is converted into an appropriate additive according to a conventional method such as 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, carboxymethylcellulose calcium, carboxymethylcellulose Mix appropriately with disintegrants such as sodium, starch, corn starch, sodium alginate, lubricants such as talc, magnesium stearate, sodium stearate, fillers or diluents such as calcium carbonate, sodium carbonate, calcium phosphate, sodium phosphate Shi It can be produced. Tablets and the like may be subjected to sugar coating, gelatin, enteric coating, film coating, and the like using a coating agent such as hydroxypropylmethylcellulose, sucrose, polyethylene glycol, titanium oxide or the like, if necessary.

  When the anticancer agent of the present invention is a liquid preparation such as an injection, eye drop, nasal drop, inhalant, spray, lotion, syrup, solution, suspension, emulsion, etc., the metal complex is purified. Water, suitable buffer solution such as phosphate buffer, physiological saline solution, Ringer solution, physiological salt solution such as lock solution, vegetable oil such as cocoa butter, sesame oil, olive oil, mineral oil, higher alcohol, higher fatty acid, ethanol, etc. In an organic solvent or the like, if necessary, an emulsifier such as cholesterol, a suspension agent such as gum arabic, a dispersion aid, a wetting agent, a surfactant such as polyoxyethylene hydrogenated castor oil type, polyethylene glycol type, Solubilizers such as sodium phosphate, stabilizers such as sugar, sugar alcohol and albumin, preservatives such as parabens, isotonic agents such as sodium chloride, glucose and glycerin, buffers, painless It can be prepared as a sterilized aqueous solution, non-aqueous solution, suspension, liposome, emulsion, etc. by appropriately adding an agent, adsorption inhibitor, moisturizer, antioxidant, colorant, sweetener, flavor, fragrance, etc. . At this time, the injection preferably has a physiological pH, specifically a pH range of 6-8.

  When the anticancer agent of this invention is a semi-solid preparation such as a lotion, cream, ointment, the metal complex is a fat, fatty oil, lanolin, petrolatum, paraffin, wax, plaster, resin, plastic, glycols It can be produced by appropriately mixing with higher alcohol, glycerin, water, emulsifier, suspending agent and the like.

  The anticancer agent of the present invention may be administered in the body by enclosing the anticancer agent of the present invention in a carrier such as a liposome, for example, a known DDS technique. In this case, if a carrier that specifically recognizes cells at the target site is used, the contrast agent of the present invention can be efficiently delivered to the target site.

(3) Content, dosage, etc.
The content of the metal complex contained in the anticancer agent of this invention, the dosage form may be adjusted arbitrarily by such dosage to severity and purposes. In addition, the dose of the anticancer agent of the present invention may be appropriately determined by a doctor according to conditions such as the age, sex, weight, symptoms, and administration route of the patient.

  The anticancer agent of the present invention may be combined with known chemotherapy, surgical treatment, radiation therapy, hyperthermia or immunotherapy.

  Hereinafter, the present invention will be described in more detail based on examples. The claims of the present invention are not limited in any way by the following examples.

1. Production of heterocyclic compound H-dpaq ^{H, ester} , which is a heterocyclic compound constituting the anticancer agent of the present invention, was synthesized along the synthetic route of FIG. In addition, in order to make it easy to understand, in the following description, the same symbol as FIG. 1 was used about the same compound.

(1) Synthesis of 6-hydroxy-8-nitroquinoline (hereinafter abbreviated as compound b) 4-amino-3-nitrophenol 5.0 g (compound a, 32 mmol), concentrated hydrochloric acid 40 mL and concentrated phosphoric acid in a reaction vessel After adding 15 g and heating to 80 ° C., 6.5 mL (97 mmol) of acrolein was slowly added. After reacting for 4 hours, the reaction solution was allowed to cool to room temperature, distilled water was added, and the mixture was filtered through Celite (Celite Corporation). The filtrate was neutralized with aqueous ammonia to obtain a red precipitate. The precipitate was collected by filtration and dried in vacuo to give a red solid (yield: 3.0 g, yield: 49%).

(2) Synthesis of Ethyl [(8-nitroquino-6-yloxy) acetate] (hereinafter abbreviated as compound c) Compound b 2.8 g (15 mmol) and K ₂ CO ₃ 4.1 g (29 mmol) in a reaction vessel Was added under anaerobic conditions, and 100 mL of dehydrated acetone was added. Further, 3.7 g (22 mmol) of ethyl bromoacetate was added dropwise and stirred overnight at room temperature. The reaction solution was filtered through Celite, and the filtrate was concentrated and then purified with an alumina column to obtain a pale yellow solid (yield: 1.0 g, yield: 25%).

(3) Synthesis of Ethyl [(8-aminoquino-6-yloxy) acetate] (hereinafter abbreviated as Compound d) Compound c 0.96 g (3.5 mmol) and 10% Pd-C 0.096 g were added to a reaction vessel, Under a nitrogen atmosphere, 40 mL of dichloromethane and 80 mL of ethanol were added and dissolved, and then the mixture was stirred overnight under a hydrogen atmosphere. The reaction solution was filtered through Celite, and the filtrate was concentrated to obtain a pale yellow solid (yield: 0.84 g, yield: 98%).

(4) Synthesis of H-dpaq ^{H, ester} (Compound e) Add 0.80 g (3.3 mmol) of compound d and 0.41 g (3.9 mmol) of Na ₂ CO _{3 to the} reaction vessel, put under nitrogen atmosphere, and add 50 mL of dehydrated acetonitrile. Dissolved in. The reaction vessel was stirred at 0 ° C. with an ice bath, and 0.34 mL (3.9 mmol) of bromoacetyl bromide was added dropwise. Twenty minutes after the start of dropping, the resulting pale yellow solid was dissolved in dichloromethane. The reaction mixture was filtered through Celite, and the filtrate was concentrated to give an orange solid.

Add the obtained N-acetylbromide-N-2-quinoline-2-carboxamide, 0.41 g (3.9 mmol) of Na ₂ CO ₃ and 0.78 g (3.9 mmol) of 2,2-dipicolylamine to the reaction vessel, and put under nitrogen atmosphere. After dissolving in 50 mL of dehydrated acetonitrile, the mixture was stirred overnight. The reaction solution was filtered through Celite, and the filtrate was concentrated and purified by an alumina column to obtain a pale yellow solid (yield: 0.95 g, yield: 60%). This compound was identified from the measurement results of nuclear magnetic resonance spectroscopy ( ¹ H NMR and ¹³ C NMR) and Fourier transform infrared spectrophotometer (FT-IR). The results are shown below.

¹ H NMR (500 MHz, CDCl ₃ ): 1.30 (t, J = 7.2 Hz, CH ₂ CH ₃ , 3H), 2.84 (s, -CH ₃ , 3H), 3.51 (s, CH ₂ CO, 2H), 3.98 (s, CH ₂ Py, 4H), 4.26 (q, J = 7.2 Hz, -CH ₂ CH ₃ , 2H), 4.73 (s, CH ₂ , 2H), 6.78 (d, J = 2.6 Hz, Qu7, 1H), 7.15 (dd, J = 7.6, 5.2 Hz, Py5, 2H), 7.35 (d, J = 8.6 Hz, Qu3, 1H), 7.62 (dd, J = 7.6, 7.6 Hz, Py4, 2H), 7.96 (d, J = 8.6 Hz, Qu4, 1H), 7.97 (t, J = 7.6 Hz, Py3, 2H), 8.52 (d, J = 5.2 Hz, Py6, 2H), 8.59 (d, J = 2.6 Hz, . Qu5, 1H), 11.66 ( s, -NHCO-, 1H) 13 C NMR (125 MHz, CDCl 3): 14.4 (CH 2 CH 3), 25.1 (CH 3), 59.7 (-CH 2 CO-), 61.3 (-CH ₂ Py), 61.6 (CH ₂ CH ₃ ), 65.8 (-CH _2- ), 101.8 (Qu7), 108.5 (Qu5), 122.6 (Py5), 123.1 (Qu3), 123.3 (Py3), 127.0 (Qu9), 135.1 (Qu10), 135.2 (Qu8), 135.7 (Qu4), 136.8 (Py4), 149.3 (Py6), 155.0 (Qu2), 156.1 (Qu6), 158.5 (Py2), 169.0 (-CO-) FT-IR (ATR): 3300 cm ^-1 (NH (amido)), 1757 cm ^-1 (C = O (ester)), 1682 cm ^-1 (C = O (amido) ).

2. Production of Metal Complex “H-dpaq ^{H, ester} ” produced in Example 1 and “H-dpaq ^{H, H} ”, “H-dpaq ^{NO2, H} ” produced by the method described in Patent Document 3, “ H-dpaq ^{OMe, H} "was used to produce the metal complexes of this invention.

(1) Synthesis of [Mn (dpaq ^{H, H} )] ClO ₄
H-dpaq ^{H, H} 100 mg (0.26 mmol) and triethyleamine 40 μL (0.29 mmol) are dissolved in 2 mL of dehydrated acetonitrile, and Mn (ClO ₄ ) ₂ · 6H ₂ O 110 mg (0.31 mmol) is dehydrated in 0.5 mL of dehydrated acetonitrile. The solution dissolved in was added dropwise under a nitrogen atmosphere and stirred overnight. The resulting precipitate was collected with a membrane filter and dried in vacuo to give a pale yellow solid (yield: 126 mg, yield: 90%). This compound was identified from the measurement results of an elemental analyzer, a Fourier transform infrared spectrophotometer (FT-IR), and a mass spectrometer (electrospray method). The results are shown below.

Anal. Calcd for [Mn (dpaq ^{H, H} )] (ClO ₄ ) (H ₂ O) _0.8 : C, 50.11; H, 3.95; N, 12.70. Found: C, 50.19; H, 3.81; N, 12.74. Selected IR frequencies (cm ^-1 , FT-ATR): 1541 (CO). Electronic absorption spectrum in CH ₃ CN (nm (M ^-1 cm ^-1 )): 375 (4500), 262 (37100). Electronic absorption spectrum in MES buffer (pH 7.2, 5% DMSO) (nm (M ^-1 cm ^-1 )): 309 (5300), 241 (23800). ESI-MS, positive mode: m / z 437.08 [Mn (dpaq ^{H, H} )] ⁺ .

(2) Synthesis of [Mn (dpaq ^{NO2, H} )] ClO ₄
^{H-dpaq NO2, H 100 mg} (0.23 mmol) and triethyleamine 34 μL of (0.25 mmol) was dissolved in dehydrated acetonitrile _{3.5 mL, Mn (ClO 4)} 2 · 6H 2 O 99 mg (0.28 mmol) of dehydrated acetonitrile 0.5 mL The solution dissolved in was added dropwise under a nitrogen atmosphere and stirred overnight. The resulting precipitate was collected by filtration with a membrane filter and dried under vacuum to obtain an orange solid (yield: 124 mg, yield: 93%). This compound was identified from the measurement results of an elemental analyzer, a Fourier transform infrared spectrophotometer (FT-IR), and a mass spectrometer (electrospray method). The results are shown below.

Anal. Calcd for [Mn (dpaq ^{NO2, H} )] (ClO ₄ ) (H ₂ O) _0.5 : C, 46.76; H, 3.41; N, 14.22. Found: C, 47.03; H, 3.26; N, 13.92. Selected IR frequencies (cm ^-1 , FT-ATR): 1533 (CO). Electronic absorption spectrum in CH ₃ CN (nm (M ^-1 cm ^-1 )): 428 (19100), 322 (3990), 262 (22900 ). Electronic absorption spectrum in MES buffer (pH 7.2, 5% DMSO) (nm (M ^-1 cm ^-1 )): 380 (10100), 238 (12000). ESI-MS, positive mode: m / z 482.15 [ Mn (dpaq ^{NO2, H} )] ⁺ .

(3) Synthesis of [Mn (dpaq ^{OMe, H} )] ClO ₄
H-dpaq ^{OMe, H} 100 mg (0.24 mmol) and triethyleamine 36 μL (0.26 mmol) were dissolved in 2 mL of dehydrated acetonitrile, and this was dissolved in 103 mL (0.29 mmol) of dehydrated acetonitrile with Mn (ClO ₄ ) ₂ · 6H ₂ O. The solution dissolved in 0.5 mL was added dropwise under a nitrogen atmosphere and stirred overnight. The resulting precipitate was collected with a membrane filter and dried under vacuum to obtain a yellow solid (yield: 75 mg, yield: 55%). This compound was identified from the measurement results of an elemental analyzer, a Fourier transform infrared spectrophotometer (FT-IR), and a mass spectrometer (electrospray method). The results are shown below.

Anal.Calcd for [Mn (dpaq ^{OMe, H} )] (ClO ₄ ) (H ₂ O) _0.5 : C, 50.06; H, 4.03; N, 12.16.Found: C, 50.22; H, 4.00; N, 12.35. Selected IR frequencies (cm ^-1 , FT-ATR): 1549 (CO). Electronic absorption spectrum in CH ₃ CN (nm (M ^-1 cm ^-1 )): 405 (3320), 346 (2330), 264 (33200 ). Electronic absorption spectrum in MES buffer (pH 7.2, 5% DMSO) (nm (M ^-1 cm ^-1 )): 334 (4270), 248 (29200). ESI-MS, positive mode: m / z 467.19 [ Mn (dpaq ^{OMe, H} )] ⁺ .

(4) Synthesis of [Mn (dpaq ^{H, ester} )] ClO ₄
^{H-dpaq H,} was dissolved in ^ester 100 mg (0.21 mmol) and triethyleamine 32 μL (0.25 mmol) of dehydrated ethanol _{50 mL, Mn (ClO 4)} 2 · 6H 2 O 90 mg (0.25 mmol) of dehydrated ethanol 1 The solution dissolved in mL was added dropwise under a nitrogen atmosphere and stirred overnight. The resulting precipitate was collected by filtration with a membrane filter and vacuum dried to obtain a pale yellow solid (yield: 86 mg, yield: 65%). This compound was identified from the measurement results of an elemental analyzer, a Fourier transform infrared spectrophotometer (FT-IR), and a mass spectrometer (electrospray method). The results are shown below.

ESI-MS, positive mode: m / z 539.14 [Mn (dpaq ^{H, ester} )] ⁺ . Selected IR frequencies (cm ^-1 , FT-ATR): 1733 (CO (ester)), 1626 (CO (amido)) .

3. Cytotoxicity Evaluation The activity (IC ₅₀ ) of the metal complex obtained in Example 2 against tumor cells was measured using HeLa cells, which are cancer cells derived from human cervical cancer. Note that cisplatin (manufactured by Wakken Pharmaceutical) was used as an experimental control. Specifically, it measured as follows.

  First, HeLa cells are inoculated into a flask containing Dulbecco's Modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum, 1% antibiotics, and antifungal agents, in a humidified incubator at 37 ° C in a 5% carbon dioxide atmosphere. Cultured. When the cultured cells reached 80% confluence in the flask, the cells in the flask were trypsinized, and the cells were washed with phosphate buffered saline (PBS).

After counting the number of cells contained in the buffer with a hemocytometer, a cell suspension of 5 × 10 ⁴ cells / ml was prepared using the medium used for cell culture. The cell suspension was added to a sterile 96-well microplate at 100 μL / well and cultured at 37 ° C. for 24 hours.

  Next, a serially diluted aqueous solution (containing 5% DMSO) of the metal complex and cisplatin obtained in Example 2 was prepared. 20 μL of this serially diluted aqueous solution was added to each well of the microplate, and cultured under the above conditions for 24 hours or 48 hours.

  After removing the culture medium in all wells, 10 μL of MTT aqueous solution (5 mg / mL) was added to each well, the microplate lid was closed, and the mixture was incubated at 37 ° C. for 4 hours. The lid was removed, and a 2-propanol solution containing 1% of 2N aqueous hydrochloric acid was added at 100 μL / well to dissolve formazan.

Finally, after measuring the absorbance at 570 nm with a 96-well plate reader, this was evaluated as the number of viable cells, and the 50% cell growth inhibitory concentration (IC ₅₀ ) was calculated. In calculating the IC ₅₀ value, an average value of at least five experimental values carried out in the same manner was adopted. The results are shown in Table 1 .

From Table 1 , it was confirmed that [Mn (dpaq ^{H, ester} )] ClO ₄ has higher cytotoxicity than cisplatin and is likely to be an excellent anticancer agent. [Mn (dpaq ^{H, H} )] ClO ₄ , [Mn (dpaq ^{NO2, H} )] ClO ₄ and [Mn (dpaq ^{OMe, H} )] ClO ₄ can also be used as anticancer agents although they do not reach cisplatin. It was confirmed that there is sex.

Claims (2)

The anticancer agent which contains the metal complex represented by following General formula (1) as an active ingredient.

(In the formula , M represents one of iron, manganese, and cobalt. X represents a counter ion, and n represents 1 or 2.) M is manganese, X is ClO ₄ ^- anticancer agents according to the is claim 1.

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