JP2021088511A - Anticancer drug - Google Patents

Abstract

To provide an anticancer drug that can be produced at low cost and has high anticancer activity.SOLUTION: An anticancer drug contains, as an active ingredient, a metal complex comprising an anion, which is a myo-inositol-1,2,3,4,5,6-hexakisphosphate derivative, represented by formula (1) (where m and n independently represent 0, 1 or 2, m+n is 2) and a counter cation, which is an alkali metal ion, an alkaline earth metal ion or a proton.SELECTED DRAWING: Figure 5

Description

The present invention relates to an anticancer agent containing a metal complex.

で表されるシスプラチン及びオキサリプラチン等の白金抗がん剤は、腎毒性等の副作用が強いにもかかわらず広範囲のがんによく効くため、多剤治療のペアとして、また、転移がん等の悪性腫瘍に対してよく用いられている。しかし、前記の通り、これらの白金抗がん剤には、腎毒性が高いことや耐性ができやすい等の問題があり、新規の白金抗がん剤が求められていた。 The following formula:

Platinum anticancer agents such as cisplatin and oxaliplatin represented by are effective against a wide range of cancers despite their strong nephrotoxicity and other side effects. It is often used for malignant tumors. However, as described above, these platinum anticancer agents have problems such as high nephrotoxicity and easy resistance, and new platinum anticancer agents have been demanded.

As such a novel platinum anticancer agent, Patent Document 1 describes myo-inositol-1,2,3,4,5,6-hexaxphosphate derivative, which is a sum of cisplatin structure and oxaliplatin structure. Disclosed is an anti-cancer agent containing three trinuclear platinum complexes. The anti-cancer agent disclosed in Patent Document 1 has achieved excellent anti-cancer activity and reduction of side effects, but is expected to further improve in terms of ease of manufacture and yield, resulting in cost reduction. It was desired.

Japanese Unexamined Patent Publication No. 2016-74639

As described above, although the conventional platinum anticancer agent has achieved excellent anticancer activity and reduction of side effects, cost reduction has also been desired. Therefore, an object of the present invention is to provide an anticancer agent that can be produced at low cost and has high anticancer activity.

As a result of various studies on means for solving the above-mentioned problems, the present inventors have myo-inositol-1,2,3,4,5,6-hexa having a total of two cisplatin structures and oxaliplatin structures. We have found that a metal complex containing a kissphosphate derivative as an anion has high anticancer activity and can be produced at low cost, and completed the present invention.

（式中、ｍ及びｎは、それぞれ独立に０、１又は２であり、ｍ＋ｎは２である。）
で表される、ｍｙｏ−イノシトール−１，２，３，４，５，６−ヘキサキスホスフェート誘導体であるアニオンと、アルカリ金属イオン、アルカリ土類金属イオン又はプロトンであるカウンターカチオンからなる金属錯体を有効成分として含有する抗がん剤。
（２）前記アニオンが、式（１−１）

で表されるアニオンである、前記（１）に記載の抗がん剤。
（３）前記アニオンが、式（１−２）

で表されるアニオンである、前記（１）に記載の抗がん剤。 That is, the gist of the present invention is as follows.
(1) Equation (1)

(In the equation, m and n are 0, 1 or 2, respectively, and m + n is 2.)
A metal complex composed of an anion, which is a myo-inositol-1,2,3,4,5,6-hexax phosphate derivative, and a counter cation, which is an alkali metal ion, an alkaline earth metal ion, or a proton, represented by. An anticancer drug contained as an active ingredient.
(2) The anion is the formula (1-1).

The anticancer agent according to (1) above, which is an anion represented by.
(3) The anion has the formula (1-2).

The anticancer agent according to (1) above, which is an anion represented by.

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide an anticancer agent that can be produced at low cost and has high anticancer activity.

FIG. 1 is a graph showing the amount of Pt in each organ of ddY mice intravenously administered with cis-cis and cis-oxali. FIG. 2 is a graph showing the amount of Pt in each organ of ddY mice orally administered with cis-cis and cis-oxali. FIG. 3A is a graph showing the amount of Pt (dose%) in each organ of ddY mice intravenously administered with cis-cis and cis-oxali. FIG. 3B is a graph showing the amount of Pt (dose%) in each organ of ddY mice intravenously administered with cisplatin. FIG. 4A is a graph showing changes in blood urea nitrogen (BUN) before and after administration of control, cisplatin, cis-oxali, oxali-cis and oxali-oxali. FIG. 4B is a graph showing changes in creatinine before and after administration of controls, cisplatin, cis-oxali, oxali-cis and oxali-oxali. FIG. 5 is a graph showing the tumor volume ratio of the control group and each test compound administration group. FIG. 6 is a graph showing the body weight ratios of the control group and each test compound administration group. FIG. 7A is a graph showing the tumor volume ratio of the control group, the cis-cis administration group, and the cis-oxali administration group, and FIG. 7B is a graph showing the tumor volume ratio of the control group and the cisplatin administration group. FIG. 8A is a graph showing the body weight ratios of the control group, the cis-cis administration group, and the cis-oxali administration group, and FIG. 8B is a graph showing the body weight ratios of the control group and the cisplatin administration group.

Hereinafter, preferred embodiments of the present invention will be described in detail.

The present invention presents an anion, which is a myo-inositol-1,2,3,4,5,6-hexakisphosphate derivative, and a counter cation. The present invention relates to an anticancer agent containing a metal complex composed of the above as an active ingredient.

（式中、ｍ及びｎは、それぞれ独立に０、１又は２であり、ｍ＋ｎは２である。）
で表される、ｍｙｏ−イノシトール−１，２，３，４，５，６−ヘキサキスホスフェート誘導体であるアニオンと、アルカリ金属イオン、アルカリ土類金属イオン又はプロトンであるカウンターカチオンからなる。本発明の金属錯体は塩の形態で存在する。本発明の金属錯体は、シスプラチン構造とオキサリプラチン構造とを合計で２つ有する二核白金錯体であり、リン酸基（−ＯＰＯ_３ ^２−）を多く有しており、骨のヒドロキシアパタイトと強く吸着する。そのため、本発明の抗がん剤は骨へのドラッグデリバリーシステム（ＤＤＳ）に利用することが期待できる。 The metal complex of the present invention has the formula (1).

(In the equation, m and n are 0, 1 or 2, respectively, and m + n is 2.)
It is composed of an anion which is a myo-inositol-1,2,3,4,5,6-hexax phosphate derivative represented by, and a counter cation which is an alkali metal ion, an alkaline earth metal ion or a proton. The metal complex of the present invention exists in the form of a salt. Metal complexes of the present invention is a two having binuclear platinum complex cisplatin structure and oxaliplatin structure in total, has many phosphate groups (-OPO 3 ^_2-), strongly hydroxyapatite bone Adsorb. Therefore, the anticancer agent of the present invention can be expected to be used in a drug delivery system (DDS) to bone.

In addition, the metal complex of the present invention has anticancer activity comparable to that of a trinuclear platinum complex having a total of three cisplatin and oxaliplatin structures. Therefore, by using the metal complex of the present invention, an anticancer agent having anticancer activity comparable to that of the trinuclear platinum complex can be produced at low cost.

In the metal complex of the present invention, the counter cation is an alkali metal ion, an alkaline earth metal ion or a proton. The alkali metal ion is not particularly limited, and examples thereof include sodium ion and potassium ion. The alkaline earth metal ion is not particularly limited, and examples thereof include calcium ion and magnesium ion. Among these, alkali metal ions are preferable as counter cations, and sodium ions are more preferable.

In the metal complex of the present invention, the anion is preferably represented by the following formulas (1-1) to (1-4). The complexes of formulas (1-3) and (1-4) contain isomers derived from the symmetry of IP6.

In the metal complex of the present invention, the anion is more preferably represented by the above formulas (1-1) and (1-2). The metal complex containing an anion represented by the formulas (1-1) and (1-2) has a symmetric structure of myo-inositol-1,2,3,4,5,6-hexax phosphate at the time of manufacture. Since it can be obtained as a single compound, it can be easily produced.

The metal complex of the present invention can be produced by using a commercially available starting material or a starting material obtained by a known synthetic method by the method described in the synthetic example described later or a method similar thereto. Further, the metal complex of the present invention can be isolated and purified by known means such as concentration, solvent extraction, recrystallization and the like.

The metal complex of the present invention may be a hydrate, an anhydrous product, a solvate product, or a non-solvate product. Further, the metal complex of the present invention may be a crystal, a single crystal form, or a mixture of crystal forms. The crystal can be produced by a crystallization method known per se.

The anticancer agent of the present invention contains the above-mentioned metal complex as an active ingredient. The anticancer agent of the present invention contains at least one of the above-mentioned metal complexes. The metal complex may be only one kind or a mixture of two or more kinds. The metal complex preferably contains anions represented by formulas (1-1) to (1-4), and more preferably represented by formulas (1-1) and (1-2). It contains anions.

The cancer to which the anticancer agent of the present invention is applied is not particularly limited, and is, for example, leukemia, malignant melanoma, malignant lymphoma, digestive organ cancer, lung cancer, esophageal cancer, gastric cancer, colon cancer, and the like. Rectal cancer, colon cancer, urinary tract tumor, bile sac cancer, bile duct cancer, biliary tract cancer, breast cancer, kidney cancer, liver cancer, pancreatic cancer, testicle tumor, maxillary cancer, tongue cancer, lips Cancer, oral cancer, pharyngeal cancer, laryngeal cancer, ovarian cancer, uterine cancer, prostate cancer, thyroid cancer, brain tumor, capsicum sarcoma, hemangiomas, true polyemia, neuroblastoma, retinal bud Examples include tumors, myelomas, bladder tumors, sarcomas, osteosarcomas, myomas, skin cancers, basal cell cancers, skin appendage cancers, skin metastatic cancers, and cutaneous melanomas.

The administration form of the anticancer agent of the present invention may be oral administration, or may be parenteral administration such as intravenous, intramuscular, subcutaneous or intradermal injection, rectal administration, and transmucosal administration. Examples of the formulation form suitable for oral administration include tablets, pills, granules, powders, capsules, liquids, suspensions, emulsions, syrups and the like. Examples of the pharmaceutical form suitable for parenteral administration include external preparations such as injections, infusions, nasal drops, sprays, inhalants and suppositories, or ointments, creams, powdered coatings and liquid coatings. , Percutaneous absorption preparations such as patches and the like. The preferred administration form, formulation form and the like are appropriately selected by the doctor according to the age, sex, constitution, symptom, treatment time and the like of the patient.

The anticancer agent of the present invention may be a solid preparation such as tablets, pills, powders, powders and granules. This solid preparation can be produced by appropriately mixing the metal complex of the present invention and a raw material for a solid preparation according to a conventional method. Raw materials for solid preparations include, for example, lactose, sucrose, D-mannitol, corn starch, synthetic or natural gum, excipients such as crystalline cellulose, starch, hydroxypropyl cellulose, hydroxypropyl methylcellulose, gum arabic, gelatin, etc. Binders such as polyvinylpyrrolidone, disintegrants such as carbosikimethylcellulose calcium, carbosikimethylcellulose sodium, starch, corn starch, sodium alginate, lubricants such as talc, magnesium stearate, sodium stearate, calcium stearate, calcium carbonate, carbonic acid. Examples thereof include fillers such as sodium, calcium phosphate, and sodium phosphate. If necessary, the anticancer agent of the present invention such as tablets may be coated with sugar, gelatin, enteric coating, film coating or the like using hydroxypropylmethyl cellulose, sucrose, polyethylene glycol, titanium oxide or the like. ..

The anticancer agent of the present invention may be a liquid preparation such as an injection, an eye drop, a nasal drop, an inhalant, a spray, a lotion, a syrup, a liquid, a suspension, or an emulsion. This liquid preparation can be produced by appropriately mixing the metal complex of the present invention and the raw material for the liquid preparation according to a conventional method. Raw materials for liquid preparations include, for example, purified water, phosphate buffer, physiological saline, ringer solution, rock solution, vegetable oil such as cacao butter, sesame oil, olive oil, mineral oil, higher alcohol, higher fatty acid, ethanol and the like. Organic solvents, emulsifiers such as cholesterol, suspending agents such as gum arabic, dispersion aids, infiltrates, surfactants such as polyoxyethylene hydrogenated castor oil and polyethylene glycol, solubilizers such as sodium phosphate, sugars , Stabilizers such as sugar alcohols and albumins, preservatives such as parabens, tonicity agents such as sodium chloride, glucose and glycerin, buffers, soothing agents, anti-adsorption agents, moisturizers, antioxidants, colorants , Sweeteners, flavors, aromatic substances and the like. When the anticancer agent of the present invention is an injection, its pH is preferably about 6 to 8.

The anticancer agent of the present invention may be a semi-solid preparation such as a lotion, a cream, or an ointment. This semi-solid preparation can be produced by appropriately mixing the metal complex of the present invention and the raw material for the semi-solid preparation according to a conventional method. Examples of the raw material for the semi-solid preparation include fat, fatty oil, lanolin, vaseline, paraffin, wax, ointment, resin, glycols, higher alcohol, glycerin, water, emulsifier, suspending agent and the like.

The content of the metal complex in the anticancer agent of the present invention may vary depending on the administration form, severity, desired dose, etc., but is generally about 0.001 to 80% by weight, preferably about 0.001 to 80% by weight. Is about 0.1 to 50% by weight.

The dose of the anticancer agent of the present invention is appropriately determined according to conditions such as the age, sex, body weight, symptoms and administration route of the patient. The daily dose of the active ingredient for an adult is generally about 1 μg / kg to 1000 mg / kg, preferably about 10 μg / kg to 10 mg / kg. The anticancer agent of the present invention may be administered once a day, or may be administered in several divided doses (for example, about 2 to 4 times) a day. The administration of the anticancer agent of the present invention may be used in combination with known chemotherapy, surgical treatment, radiation therapy, hyperthermia, immunotherapy and the like.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the technical scope of the present invention is not limited to these examples.

The meanings of the abbreviations of the raw materials used in the following synthetic examples and test examples are as follows.
dach: 1R, 2R-1,2-Cyclohexanediamine IP6: myo-inositol-1,2,3,4,5,6-hexax phosphate anion (composition formula: C ₆ H ₆ O ₂₄ P ₆ )
AtC3: Anthracene-CH ₂ NH (CH ₂ ) ₃ NH ₂

The synthesized metal complexes were ^{identified by elemental analysis, 31} P-NMR and ¹⁹⁵ Pt-NMR. The equipment used is described below.
Elemental analysis: Leco CHN 900
³¹ P-NMR: JEOL-ECA600 (25 ° C, 243 MHz)
¹⁹⁵ Pt-NMR: Varian-VXR300S (25 ° C, 64.497 MHz)

Ｋ_２ＰｔＣｌ_４（２．０ｇ、４．８ｍｍｏｌ）を水（１０ｍｌ）に溶解させ、この溶液にＫＩ（３．３ｇ、１９．８ｍｍｏｌ）を加えて、５０℃の湯浴で温めながら５分間撹拌した。この溶液を、１．０ｍｏｌ／Ｌアンモニア水（９．６ｍＬ、９．６ｍｍｏｌ）に加え、室温で３時間静置後、沈殿物をろ取し、熱水、エタノール、ジエチルエーテルの順で洗浄後、真空乾燥して、Ｐｔ（ＮＨ_３）_２Ｉ_２を黄色粉末として得た（収率：９７％）。
元素分析：Ｈ_６Ｎ_２Ｉ_２Ｐｔ
計算値 Ｃ：０．００、Ｈ：１．２５、Ｎ：５．８０
実測値 Ｃ：０．００、Ｈ：１．０９、Ｎ：５．４９ <Synthesis of metal complex>
(1) Synthesis of _{Pt (NH 3} ) ₂ I ₂ (cisPt-I ₂₎

Dissolve K ₂ PtCl ₄ (2.0 g, 4.8 mmol) in water (10 ml), add KI (3.3 g, 19.8 mmol) to this solution, and stir for 5 minutes while warming in a hot water bath at 50 ° C. did. This solution is added to 1.0 mol / L aqueous ammonia (9.6 mL, 9.6 mmol), allowed to stand at room temperature for 3 hours, the precipitate is collected by filtration, and washed in the order of hot water, ethanol, and diethyl ether. , Vacuum dried to give Pt (NH ₃ ) ₂ I ₂ as a yellow powder (yield: 97%).
Elemental analysis: H ₆ N ₂ I ₂ Pt
Calculated values C: 0.00, H: 1.25, N: 5.80
Measured values C: 0.00, H: 1.09, N: 5.49

Ｋ_２ＰｔＣｌ_４（２．０ｇ、４．８ｍｍｏｌ）を水（１０ｍｌ）に溶解させ、この溶液にＫＩ（３．３ｇ、１９．８ｍｍｏｌ）を加えて、５０℃の湯浴で温めながら５分間撹拌した。この溶液を、ｄａｃｈ（０．５５ｇ、４．８ｍｍｏｌ）を含む溶液に加え、室温で３時間静置した後、沈殿物をろ取し、熱水、エタノール、ジエチルエーテルの順で洗浄後、真空乾燥して、Ｐｔ（ｄａｃｈ）Ｉ_２を黄色粉末として得た（収率：９９％）。
元素分析：Ｃ_６Ｈ_１４Ｎ_２Ｉ_２Ｐｔ
計算値 Ｃ：１２．７９、Ｈ：２．４９、Ｎ：４．９７
実測値 Ｃ：１２．９２、Ｈ：２．３７、Ｎ：５．１３ (2) Synthesis of Pt (dach) I ₂ (oxalipt-I ₂ )

Dissolve K ₂ PtCl ₄ (2.0 g, 4.8 mmol) in water (10 ml), add KI (3.3 g, 19.8 mmol) to this solution, and stir for 5 minutes while warming in a hot water bath at 50 ° C. did. This solution is added to a solution containing dach (0.55 g, 4.8 mmol), allowed to stand at room temperature for 3 hours, the precipitate is collected by filtration, washed with hot water, ethanol, and diethyl ether in this order, and then vacuumed. Drying gave Pt (dach) I ₂ as a yellow powder (yield: 99%).
Elemental analysis: C ₆ H ₁₄ N ₂ I ₂ Pt
Calculated values C: 12.79, H: 2.49, N: 4.97
Measured values C: 12.92, H: 2.37, N: 5.13

硝酸銀（１．３６ｇ、８．０ｍｍｏｌ）を水（８０ｍｌ）に溶解させ、この溶液にＰｔ（ＮＨ_３）_２Ｉ_２（１．９２ｇ、４．０ｍｍｏｌ）を溶解させたジメチルアセトアミド溶液（２０ｍｌ）を加えて遮光し、２４時間撹拌した。この溶液をろ過してヨウ化銀を除去し、ＩＰ６・１２Ｎａ（４．４ｇ、４．０ｍｍｏｌ）を含む水溶液（１２ｍｌ）を加え、室温で５時間撹拌した。得られた溶液をろ過し、ろ液を減圧濃縮し、メタノールを加え、茶色沈殿物をろ取し、真空乾燥して、Ｐｔ（ＮＨ_３）_２ＩＰ６・１０Ｎａ・ＭｅＯＨ・１４Ｈ_２Ｏを得た（収率：９３％）。
元素分析：Ｃ_７Ｈ_４２Ｎ_２ＰｔＰ_６Ｏ_３８Ｎａ_１０
Ｐｔ（ＮＨ_３）_２ＩＰ６・１０Ｎａ・ＭｅＯＨ・１４Ｈ_２Ｏ
計算値 Ｃ：６．０１、Ｈ：３．１４、Ｎ：２．００
実測値 Ｃ：６．０７、Ｈ：２．７７、Ｎ：１．８３
^３１Ｐ ＮＭＲ（Ｄ_２Ｏ，８５％ Ｈ_３ＰＯ_４基準）δ（ｐｐｍ）＋７．６６（１Ｐ），＋７．５１（１Ｐ），＋３．５８（４Ｐ）
^１９５Ｐｔ ＮＭＲ（Ｄ_２Ｏ，Ｐｔ（ｅｎ）_２Ｃｌ_２基準）δ（ｐｐｍ）−２８２１ (3) _{Synthesis of Pt (NH 3} ) ₂ IP6 ・ 10Na (cisPt-IP6)

A dimethylacetamide solution (20 ml) in which silver nitrate (1.36 g, 8.0 mmol) was dissolved in water (80 ml) and Pt (NH ₃ ) ₂ I ₂ (1.92 g, 4.0 mmol) was dissolved in this solution was added. In addition, it was shielded from light and stirred for 24 hours. This solution was filtered to remove silver iodide, an aqueous solution (12 ml) containing IP6.12Na (4.4 g, 4.0 mmol) was added, and the mixture was stirred at room temperature for 5 hours. The obtained solution was filtered, the filtrate was concentrated under reduced pressure, methanol was added, the brown precipitate was collected by filtration, and dried in vacuum to obtain Pt (NH ₃ ) ₂ IP6 ・ 10Na ・ MeOH ・ 14H ₂ O. (Yield: 93%).
Elemental analysis: C ₇ H ₄₂ N ₂ PtP ₆ O ₃₈ Na ₁₀
Pt (NH ₃ ) ₂ IP6 ・ 10Na ・ MeOH ・ 14H ₂ O
Calculated values C: 6.01, H: 3.14, N: 2.00
Measured values C: 6.07, H: 2.77, N: 1.83
³¹ P NMR (D ₂ O, 85% H ₃ PO ₄ standard) δ (ppm) +7.66 (1P), +7.51 (1P), +3.58 (4P)
_{^{195 Pt NMR (D 2 O,}} Pt (en) 2 Cl 2 standard) δ (ppm) -2821

硝酸銀（０．３４ｇ、２．０ｍｍｏｌ）を水（２０ｍｌ）に溶解させ、Ｐｔ（ＮＨ_３）_２Ｉ_２（０．４８ｇ、１．０ｍｍｏｌ）を溶解させたジメチルアセトアミド溶液（５ｍｌ）を加えて遮光し、６０℃で２４時間撹拌した。この溶液をろ過してヨウ化銀を除去し、Ｐｔ（ＮＨ_３）_２ＩＰ６・１０Ｎａ（１．４０ｇ、１．０ｍｍｏｌ）を含む水溶液（５ｍｌ）を加え、６０℃で３晩撹拌した。得られた溶液をろ過して、ろ液を減圧濃縮し、得られた残渣にメタノールを加えた後、茶色沈殿物をろ取し、真空乾燥して、（Ｐｔ（ＮＨ_３）_２・Ｐｔ（ＮＨ_３）_２）ＩＰ６・８Ｎａ・ＭｅＯＨ・１１Ｈ_２Ｏを得た（収率：７８％）。
元素分析：Ｃ_７Ｈ_４４Ｎ_４Ｏ_３６Ｐｔ_２Ｐ_６Ｎａ_８
計算値 Ｃ：５．５３、Ｈ：２．８９、Ｎ：３．６８
実測値 Ｃ：５．８１、Ｈ：３．００、Ｎ：３．７１
^３１Ｐ ＮＭＲ（Ｄ_２Ｏ，８５％ Ｈ_３ＰＯ_４基準）δ（ｐｐｍ）＋７．６６（１Ｐ），＋７．５１（１Ｐ），＋３．５８（４Ｐ）
^１９５Ｐｔ ＮＭＲ（Ｄ_２Ｏ，Ｐｔ（ｅｎ）_２Ｃｌ_２基準）δ（ｐｐｍ）−２８２１ (4) Synthesis of (Pt (NH ₃ ) ₂ · Pt (NH ₃ ) ₂ ) IP6.8Na (cis-cis)

Silver nitrate (0.34 g, 2.0 mmol) is dissolved in water (20 ml), _{and dimethylacetamide solution (5 ml) in which Pt (NH 3} ) ₂ I ₂ (0.48 g, 1.0 mmol) is dissolved is added to block light. Then, the mixture was stirred at 60 ° C. for 24 hours. This solution was filtered to remove silver iodide _{, an aqueous solution (5 ml) containing Pt (NH 3} ) ₂ IP6 / 10Na (1.40 g, 1.0 mmol) was added, and the mixture was stirred at 60 ° C. for 3 nights. The obtained solution was filtered, the filtrate was concentrated under reduced pressure, methanol was added to the obtained residue, the brown precipitate was collected by filtration, and the mixture was vacuum dried to (Pt (NH _{3 ) 2.} Pt (Pt (NH 3) 2. Pt). _{NH 3) 2) IP6 · 8Na} · MeOH · 11H was obtained ₂ O (yield: 78%).
Elemental analysis: C ₇ H ₄₄ N ₄ O ₃₆ Pt ₂ P ₆ Na ₈
Calculated values C: 5.53, H: 2.89, N: 3.68
Measured values C: 5.81, H: 3.00, N: 3.71
³¹ P NMR (D ₂ O, 85% H ₃ PO ₄ standard) δ (ppm) +7.66 (1P), +7.51 (1P), +3.58 (4P)
_{^{195 Pt NMR (D 2 O,}} Pt (en) 2 Cl 2 standard) δ (ppm) -2821

硝酸銀（０．３４ｇ、２．０ｍｍｏｌ）を水（２０ｍｌ）に溶解させ、この溶液にＰｔ（ｄａｃｈ）Ｉ_２（０．５６ｇ、１．０ｍｍｏｌ）を溶解させたジメチルアセトアミド溶液（５ｍｌ）を加えて遮光し、６０℃で２４時間撹拌した。この溶液をろ過してヨウ化銀を除去し、Ｐｔ（ＮＨ_３）_２ＩＰ６・１０Ｎａ（１．４ｇ、１．０ｍｍｏｌ）を含む水溶液（５ｍｌ）を加え、６０℃で３晩撹拌した。得られた溶液をろ過して、ろ液を減圧濃縮し、得られた残渣にメタノールを加えた後、茶色沈殿物をろ取し、真空乾燥して、（Ｐｔ（ＮＨ_３）_２・Ｐｔ（ｄａｃｈ））ＩＰ６・８Ｎａ・ＭｅＯＨ・８Ｈ_２Ｏを得た（収率：８７％）。
元素分析：Ｃ_１３Ｈ_４６Ｎ_４Ｐｔ_２Ｐ_６Ｏ_３３Ｎａ_８
計算値 Ｃ：１０．０９、Ｈ：２．９８、Ｎ：３．６２
実測値 Ｃ：１０．４６、Ｈ：２．９８、Ｎ：３．３９
^３１Ｐ ＮＭＲ（Ｄ_２Ｏ，８５％ Ｈ_３ＰＯ_４基準）δ（ｐｐｍ）＋７．８０（２Ｐ），＋３．６０（４Ｐ）
^１９５Ｐｔ ＮＭＲ（Ｄ_２Ｏ，Ｐｔ（ｅｎ）_２Ｃｌ_２基準）δ（ｐｐｍ）−２８２１，−３０２２ (5) Synthesis of (Pt (NH ₃ ) ₂ , Pt (dach)) IP6.8Na (cis-oxali)

Silver nitrate (0.34 g, 2.0 mmol) is dissolved in water (20 ml), _{and a dimethylacetamide solution (5 ml) in which Pt (dach) I 2} (0.56 g, 1.0 mmol) is dissolved is added to this solution. It was shielded from light and stirred at 60 ° C. for 24 hours. This solution was filtered to remove silver iodide _{, an aqueous solution (5 ml) containing Pt (NH 3} ) ₂ IP6 / 10Na (1.4 g, 1.0 mmol) was added, and the mixture was stirred at 60 ° C. for 3 nights. The obtained solution was filtered, the filtrate was concentrated under reduced pressure, methanol was added to the obtained residue, the brown precipitate was collected by filtration, and the mixture was vacuum dried to (Pt (NH _{3 ) 2.} Pt (Pt (NH 3) 2. Pt). dach)) IP6 · 8Na · obtain a MeOH · 8H ₂ O (yield: 87%).
Elemental analysis: C ₁₃ H ₄₆ N ₄ Pt ₂ P ₆ O ₃₃ Na ₈
Calculated values C: 10.09, H: 2.98, N: 3.62
Measured values C: 10.46, H: 2.98, N: 3.39
³¹ P NMR (D ₂ O, 85% H ₃ PO ₄ standard) δ (ppm) +7.80 (2P), +3.60 (4P)
_{^{195 Pt NMR (D 2 O,}} Pt (en) 2 Cl 2 standard) δ (ppm) -2821, -3022

Further, referring to the examples of JP-A-2016-74639, a mononuclear, dinuclear or trinuclear platinum complex having a total of one, two or three cisplatin structures and / or oxaliplatin structures was synthesized. did. These dinuclear and trinuclear complexes contain isomers derived from IP6 symmetry.

Mononuclear platinum complex

Dinuclear platinum complex

Trinuclear platinum complex

<Evaluation of in vitro cell proliferation suppression by MTT assay>
Using 39 types of human cancer cells, the in vitro anticancer activity of metal complexes such as cis-cis and cis-oxali was measured. Specifically, cancer cells were seeded on a 96-well plate, the next day, a sample solution (5 doses, 10-4 to 10-8 at 1 log intervals) was added, and after culturing for 2 days, cell proliferation was carried out by sulforhodamine B. It was measured by colorimetric quantification. The measurement results were input to the computer and the data was processed. Table 1 shows the 50% cell proliferation inhibitory concentration (IC ₅₀ ) (average value of 39 types of human cancer cells) of each metal complex.

The 39 types of human cancer cells used were breast cancer (HBC-4, BSY-1, HBC-5, MCF-7, MDA-MB-231), and central nervous system tumors (U251, SF-268, SF-295). , SF-539, SNB-75, SNB-78), colorectal cancer (HCC2998, KM-12, HT-29, HCT-15, HCT-116), lung cancer (NCI-H23, NCI-H226, NCI-H522) , NCI-H460, A549, DMS273, DMS114), melanoma (LOX-IMVI), ovarian cancer (OVCAR-3, OVCAR-4, OVCAR-5, OVCAR-8, SK-OV-3), kidney cancer ( RXF-631L, ACHN), gastric cancer (St-4, MKN1, MKN7, MKN28, MKN45, MKN74), prostate cancer (DU-145, PC-3).

As shown in Table 1, the dinuclear platinum complex of the present invention showed comparable high anticancer activity as compared to the corresponding trinuclear platinum complex.

<Pt distribution experiment>
In order to confirm the accumulation of cis-cis and cis-oxali in organs in vivo and the presence or absence of accumulation in specific organs, male ddY mice were used to each organ when intravenously or orally administered. The distribution of was examined.

Intravenous administration cis-cis, cis-omega or cisplatin was intravenously administered to male ddY mice (6 weeks old, n = 3). The dose was 12.4 μmol / kg for cis-cis and cis-oxali and 8.25 μmol / kg for cisplatin. 24 and 48 hours after administration, each organ was solubilized with a 25% aqueous solution of tetramethylammonium hydroxide (60% nitrate for bone), and the amount of Pt in each organ was inductively coupled plasma-Atomic. It was measured by Emission Spectrometry (ICP-AES).

Oral administration cis-cis or cis-oxali was orally administered to male ddY mice (6 weeks old, n = 3) at a dose of 248 μmol / kg, and 2 hours after administration, each organ was orally administered with a 25% tetramethylammonium hydroxide aqueous solution. It was solubilized with (60% nitrate for bone), and the amount of Pt in each organ was measured by ICP-AES.

FIG. 1 shows the amount of Pt in each organ of ddY mice intravenously administered with cis-cis and cis-oxali, and FIG. 2 shows the amount of Pt in each organ of ddY mice orally administered with cis-cis and cis-oxali. Indicates the amount of Pt. Further, FIG. 3A shows the amount of Pt (dose%) in each organ of the ddY mouse intravenously administered with cis-cis and cis-oxali, and FIG. 3B shows each organ of the ddY mouse intravenously administered with cisplatin. The amount of Pt in (dose%) is shown.

As shown in FIG. 1, when administered intravenously, both cis-cis and cis-oxali tended to accumulate in the liver, kidney, bone and spleen as compared to other organs. Further, as shown in FIG. 2, in the case of oral administration, cis-cis tends to accumulate more in the liver, kidney, bone and spleen, and in the stomach, small intestine and large intestine as compared with other organs. cis-oxali showed a tendency to accumulate more in the liver, kidneys, bones, stomach, small intestine, and large intestine than in other organs.

As shown in FIGS. 3A and 3B, cis-cis and cis-oxali showed higher proportions compared to cisplatin, especially in the liver and spleen. Therefore, the metal complex of the present invention can be expected to be used in a drug delivery system (DDS) to these organs.

<In vivo nephrotoxicity evaluation>
Male ICR mice (7 weeks old) 16 hours after fasting were intravenously administered with water (control), cisplatin, cis-oxali, oxali-cis or oxali-oxali at a dose of 33 μmol / kg. 56 hours later, fasting was started, and 16 hours later, blood was collected from a vein located behind the tibia using an animal lancet, and an index of renal function was used using i-STAT1 Analyzer and cartridge CHEM8 +. Blood urea nitrogen (BUN) and creatinine levels were measured. Since a creatinine value of 0.2 or less cannot be measured, it was set to 0.2. 4A and 4B show changes in BUN and creatinine before and after administration of controls, cisplatin, cis-oxali, oxali-cis and oxali-oxali, respectively.

As shown in FIGS. 4A and 4B, in the cisplatin-administered group, an increase in BUN and creatinine was observed as compared with the control group in which no metal complex was administered, and nephrotoxicity was confirmed. On the other hand, in the metal complex-administered group of the present invention, no increase in the values of BUN and creatinine was observed.

<In vivo anticancer experiment by oral administration>
The anticancer activity in vivo when the metal complex of the present invention cis-cis, cis-oxali and the conventional anticancer agents Pt (dach) AtC3 and oxaliplatin were orally administered was evaluated. Pt (dach) AtC3 has the following structure and was synthesized according to the examples of JP-A-2018-135297.

A cell suspension of mouse colorectal cancer-derived cell Colon-26 was prepared so that the number of cells was ^{1 × 10 6} per 100 μl of the medium. This cell suspension was injected into male BALB / c mice (5 weeks old) and the mice were inoculated with cancer cells.

Day 0 was 7 days after the day when the tumor cells were injected into the mice, fasted for 15 hours before administration, and the test compound was orally administered to the treatment group once every 2 days on the 7th, 8th, and 14th days. , Body weight and tumor volume were measured on day 21. The dose was 200 μl water / 20 g untreated (n = 3) in the control group, and 248 μmol / kg (cis-cis administration group: n = 3, cis-oxali administration group: n = 3,) in the test compound administration group. Pt (dach) AtC3 administration group: n = 4, oxaliplatin administration group: n = 3). Tumor volume was calculated as (length x width ² x 0.52) and shown by relative evaluation. FIG. 5 shows the tumor volume ratio of the control group and each test compound administration group, and FIG. 6 shows the body weight ratio of the control group and each test compound administration group. Experiments on Pt (dach) AtC3 and other test compounds are conducted separately.

In FIGS. 5 and 6, control 1 corresponds to the control group of Pt (dach) AtC3, and control 2 corresponds to the control group of other test compounds. As shown in FIGS. 5 and 6, control 1 and control 2 follow almost the same course, so that it can be said that the test results for Pt (dach) AtC3 and other test compounds are comparable.

As shown in FIG. 5, the cis-cis-administered group and the cis-oxali-administered group showed a significantly lower tumor volume ratio as compared with the conventional Pt (dach) AtC3-administered group and the oxaliplatin-administered group. It suppressed the growth of the tumor. In addition, as shown in FIG. 6, weight loss was observed after administration in the conventional Pt (dach) AtC3 administration group, but no weight loss was observed in the cis-cis and cis-oxali administration groups, and appetite was observed. No reduction occurred.

<In vivo anticancer experiment by intravenous administration>
The anticancer activity in vivo when the metal complex of the present invention cis-cis, cis-oxali and the conventional anticancer agents Pt (dach) AtC3 and cisplatin were intravenously administered was evaluated.

A cell suspension of mouse colorectal cancer-derived cell Colon-26 was prepared so that the number of cells was ^{1 × 10 6} per 100 μl of the medium. This cell suspension was injected into male BALB / c mice (5 weeks old) and the mice were inoculated with cancer cells.

Seven days after the day when the cancer cells were injected into the mice was defined as the 0th day, and for the cis-cis administration group and the cis-oxali administration group, the test compound was intravenously administered on the 0th, 7th and 14th days. Then, the body weight and tumor volume were measured. The dose was 100 μl water / 20 g untreated (n = 4) in the control group and 12.4 μmol / kg (n = 3) in the test compound administration group. For the cisplatin-administered group, the test compound was intravenously administered on the 0th day, the 3rd day, the 7th day, the 10th day and the 14th day, and the body weight and the tumor volume were measured. The dose was 100 μl water / 20 g untreated (n = 5) in the control group and 8.25 μmol / kg (n = 5) in the cisplatin-administered group. Tumor volume was calculated as (length x width ² x 0.52) and shown by relative evaluation. FIG. 7A shows the tumor volume ratio of the control group, the cis-cis administration group, and the cis-oxali administration group, and FIG. 7B shows the tumor volume ratio of the control group and the cisplatin administration group. Further, FIG. 8A shows the body weight ratios of the control group, the cis-cis administration group, and the cis-oxali administration group, and FIG. 8B shows the body weight ratios of the control group and the cisplatin administration group.

As shown in FIGS. 7A and 7B, the cis-cis-administered group and the cis-oxali-administered group showed a significantly lower tumor volume ratio and suppressed tumor growth as compared with the cisplatin-administered group. In addition, as shown in FIGS. 8A and 8B, in the cis-cis-administered group and the cis-oxali-administered group, the weight loss observed in the cisplatin-administered group was not observed, and the appetite was not decreased.

Claims (3)

Equation (1)

(In the equation, m and n are 0, 1 or 2, respectively, and m + n is 2.)
A metal complex composed of an anion, which is a myo-inositol-1,2,3,4,5,6-hexax phosphate derivative, and a counter cation, which is an alkali metal ion, an alkaline earth metal ion, or a proton, represented by. An anticancer drug contained as an active ingredient. The anion is of formula (1-1).

The anticancer agent according to claim 1, which is an anion represented by. The anion is of formula (1-2).

The anticancer agent according to claim 1, which is an anion represented by.

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