JP4838394B1 - Gold complex and pharmaceutical composition containing the same - Google Patents

Abstract

で表わされる光学活性なN-(2-ピコリノイル）スピロ[4,4]ノナン-1,6-ジアミンを配位子として有する新規金錯体および該錯体を有効成分とする悪性腫瘍治療剤を提供する。
【選択図】なしDisclosed are a novel gold complex which has strong antitumor activity and is effective at a lower dose, and a pharmaceutical composition containing the same.
[Solution]
The following general formula

And a novel gold complex having an optically active N- (2-picolinoyl) spiro [4,4] nonane-1,6-diamine as a ligand and a therapeutic agent for malignant tumors containing the complex as an active ingredient .
[Selection figure] None

Description

  The present invention relates to a novel gold complex and a pharmaceutical composition comprising the same as an active ingredient, particularly a malignant tumor therapeutic agent.

  Conventionally, the gold auranofin [I] has been used as an anti-rheumatic drug. Further, this compound [I] has an antitumor effect (see Non-Patent Document 1).

  Since then, many gold complexes have been synthesized and their antitumor activity has been investigated. Among them, gold complex [II] having N- (2-picolinoyl) -2-aminoquinoline of the following formula as a ligand has been reported to have an antitumor action (see Non-Patent Document 2).

  As platinum complexes, cisplatin [III], carboplatin [IV], oxaliplatin [V] and the like have been developed as anticancer agents and used for treatment (for example, Non-Patent Document 3 to Non-Patent Document 5). reference). The applicant of the present application has registered Japanese Patent No. 4621285 (Patent Document 1) as a gold complex with an antitumor effect. However, gold complexes have not yet been used as anticancer agents.

Japanese Patent No. 4621285

J. Med. Chem. 1986 29, 218-223 Dalton Trans 2003, 3419-3424 Nature, 1969 222 385-386 Cancer Treat Reviews 1985 12 21-33 Cancer Letters 1985 27 135-143

  Platinum complexes developed as anticancer agents have the disadvantage of having enormous side effects.

  The object of the present invention is based on a report disclosed in Non-Patent Document 2, and (R, R, R) -N- (2-picolinoyl) spiro [4,4] nonane-1,6-diamine and ( Gold complex with S, S, S) -N- (2-picolinoyl) spiro [4,4] nonane-1,6-diamine as a ligand, with strong antitumor activity and lower dosage The object is to provide a novel gold complex which is effective and therefore has reduced side effects.

  In order to achieve such an object, the present invention provides an optically active (R, R, R) -N- (2-picolinoyl) spiro [4,4] nonane-1,6- Novel gold complexes (D), (E) and the complexes having diamine and (S, S, S) -N- (2-picolinoyl) spiro [4,4] nonane-1,6-diamine as ligands The active ingredient.

Here, X in the formula represents a halogen ion such as Cl or an anion such as NO ₃ , HSO ₄ , PF ₆ , or AuCl ₄ .

  The present invention is a gold complex having N- (2-picolinoyl) spiro [4,4] nonane-1,6-diamine represented by the following three-dimensional structural formula (A) as a ligand.

  The present invention provides a gold having (R, R, R) -N- (2-picolinoyl) spiro [4,4] nonane-1,6-diamine represented by the following three-dimensional structural formula (B) as a ligand. It is a complex.

  The present invention provides a gold having (S, S, S) -N- (2-picolinoyl) spiro [4,4] nonane-1,6-diamine represented by the following three-dimensional structural formula (C) as a ligand. It is a complex.

  The present invention is the gold complex described above, which is a gold complex represented by the formula (D).

Here, X in the formula represents a halogen ion such as Cl or an anion such as NO ₃ , HSO ₄ , PF ₆ , or AuCl ₄ .

  The present invention is the gold complex described above, which is a gold complex represented by the formula (E).

Here, X in the formula represents a halogen ion such as Cl or an anion such as NO ₃ , HSO ₄ , PF ₆ , or AuCl ₄ .

  The present invention is the above gold complex in which X is chlorine.

  The present invention is a pharmaceutical composition containing the above gold complex as an active ingredient.

  This invention is a malignant tumor therapeutic agent which contains the above-mentioned gold complex as an active ingredient.

  The novel gold complexes (D) and (E) of the present invention are malignant tumors, particularly human lung cancer cells, human gastric cancer cells, human liver cancer cells, human pancreatic cancer cells, human colon cancer cells, human rectal cancer cells, It has strong antitumor activity against various malignant tumors such as human malignant melanoma cells, human bladder cancer cells, human lymphoma cells, etc., and is effective at smaller doses compared with conventional platinum complex malignant tumor therapeutic agents Therefore, side effects are relatively reduced.

It is a graph which shows the toxicity (cell growth inhibition rate (%)) at the time of making the compound (D) of this invention and oxaliplatin act on a normal hepatocyte (THLE-2).

  Embodiments of the gold complex of the present invention and a malignant tumor therapeutic agent containing the same will be described below.

  The spiro [4,4] nonane-1,6-diamine as a raw material according to the present invention has three stereoisomers of cis, cis-, cis, trans-, trans and trans- in terms of stereochemistry. However, the trans and trans isomers are sterically unable to form a complex with gold in the molecule, and are preferably cis, cis, cis, and trans isomers. Particularly preferred are cis and cis isomers. This compound has two enantiomers of (R, R, R) and (S, S, S).

  Where both enantiomers, ie (R, R, R) -cis, cis-spiro [4,4] nonane-1,6-diamine, (S, S, S) -cis, cis-spiro [4,4] Nonane-1,6-diamine is synthesized and converted into amides (B) and (C) of 2-picolinic acid, and then gold complexes (D) and (E) having this compound as a ligand are provided. The present invention has been completed.

  Since both enantiomers are synthesized by the same method, a gold complex having (R, R, R) -N- (picolinoyl) spiro [4,4] nonane-1,6-diamine (B) as a ligand (D ) Will be described.

  This compound (D) is easily synthesized by the method represented by the following reaction formula (i) with reference to Non-Patent Document 1 (J. Med. Chem. 1986 29, 218-223).

  The novel compound (B) was synthesized from (R, R, R) -cis, cis-spiro [4,4] nonane-1,6-diamine (F) according to the method of the following reaction formula (ii).

  (R, R, R) -cis, cis-spiro [4,4] nonane-1,6-diamine (F) is prepared by a known method (ACC Chan et al. Tetrahedoron. Lett., 2004, 45, 7379), for example, reaction. It can be synthesized by the method shown in formula (iii). That is, it can be synthesized from the corresponding (S, R, S) -trans, trans-spiro [4,4] nonane-1,6-diol (J) in three steps.

Here, MsCl represents methanesulfonyl chloride (CH ₃ SO ₂ Cl).

  (S, R, S) -trans, trans-spiro [4,4] nonane-1,6-diol (J) is prepared by a known method (ACC Chan et al. Tetrahedoron. Lett., 2004, 45, 7379), that is, spiro. It can be synthesized by reducing [4,4] -nonane-1,6-dione (R) with borane using known optically active (R) -CBS (M) as a catalyst (reaction formula (iv)) .

  The (R) -CBS catalyst (M) can be synthesized by a known method (E. J. Corey, R. K. Bakshi, S. Shibata J. Am. Chem. Soc., 1987, 109, 5551) according to the following reaction formula (v).

  Spiro [4,4] nonane-1,6-dione (N) as a raw material is obtained from a commercially available compound (S) according to a known method (JA Nieman, BA Keay, Synthetic Comm., 1999, 29, 3929).

  When a pharmaceutical composition containing an effective amount of the gold complex of the present invention is administered clinically, it is administered orally or parenterally. The dosage forms include tablets, dragees, pills, capsules, powders, troches, solutions, suppositories, injections, and the like, which are produced by blending pharmaceutically acceptable excipients. The pharmaceutical composition of the present invention is preferably prepared as a parenteral preparation. The following can be illustrated as an excipient | filler. Lactose, sucrose, glucose, sorbitol, mannitol, potato starch, amylopectin, other various starches, cellulose derivatives (eg, carboxymethylcellulose, hydroxyethylcellulose, etc.), gelatin, calcium stearate, magnesium stearate, polyvinyl alcohol, polyethylene glycol Wax, gum arabic, talc, titanium dioxide, olive oil, peanut oil, sesame oil, etc. Agents, stabilizers, isotonic agents, buffers and the like, and other pharmaceutically acceptable excipients.

  The therapeutic agent of the present invention can contain 0.001 to 85% by weight, preferably 0.005 to 60% by weight of the gold complex of the present invention.

  The dosage of the therapeutic agent of the present invention depends mainly on the symptoms, but is 0.005 to 200 mg, preferably 0.01 to 50 mg per adult body weight per day.

Hereinafter, the present invention will be described in more detail with reference to examples.
Example 1 Synthesis of Compound (B)
Dissolve 2.00 g of (R, R, R) -cis, cis-spiro [4,4] nonane-1,6-diamine (F) in 60 ml of 10% triethylamine in methanol and stir for 15 minutes. A solution prepared by dissolving di-t-butyl dicarbonate ((Boc) ₂ O 1.03 g, 0.36 eq.) In 60 ml of anhydrous methanol is dropped into this solution over about 1.5 hours. Stir for 2 days after dropping. The solvent is distilled off, 40 ml of water is added and extracted 5 times with 50 ml of methylene chloride. Dry over anhydrous sodium sulfate. After distilling off the solvent, the residue is separated by silica gel (60 g) chromatography. The eluent is chloroform-methanol-aqueous ammonia (92: 8: 1). 1.2 g (yield 36%) of (R, R, R) -N- (t-butoxycarbonyl) spiro [4,4] nonane-1,6-diamine (G) were obtained.
Results of infrared absorption analysis
IR (cm ^-1 ): 3294, 2960, 2890, 1699, 1506, 1475, 1446, 1390, 1367, 1250, 1172, 1089, 1043
Dissolve 0.625 g of 2-picolinic acid in 12 ml of DMF, add 0.686 g of 1-hydroxy-1H-benzotriazole (HOBt) and 0.972 g of dicarbodiimide (DCC), and stir for 1 hour under ice cooling. Next, 1.175 g of (R, R, R) -N- (t-butoxycarbonyl) spiro [4,4] nonane-1,6-diamine (G) synthesized earlier is dissolved in 15 ml of DMF, and this solution is added dropwise. . Stir overnight after the addition. The solution is filtered through celite, washed with 150 ml of ethyl acetate and the filtrate is washed 3 times with 30 ml of water and once with brine. Dry over anhydrous sodium sulfate.

After distilling off the solvent, the residue is separated by silica gel (36 g) chromatography. The eluent is hexane-ethyl acetate (1.5: 1). (R, R, R) -N- (t-butoxycarbonyl) -N`- (2-picolinoyl) -spiro [4,4] nonane-1,6-diamine (H) 1.19 g (yield 72%) was gotten.
IR (cm <sup>-1</sup> ): 3322, 2964, 2874, 1709, 1670, 1591, 1560, 1522, 1466, 1435, 1366, 1246, 1171, 1090, 1043
0.55 g of N- (t-butoxycarbonyl) -N`- (2-picolinoyl) diamine (H) is dissolved in 15 ml of methylene chloride, and 3 ml of trifluoroacetic acid (TFA) is slowly added dropwise under ice cooling. Thereafter, the mixture is stirred at room temperature for about 6 hours. Add 12 ml of benzene and distill off about half amount of solvent. After performing this operation three times, the whole amount is distilled off. The residue is dissolved in a small amount of water, desalted with an ion exchange resin, the obtained aqueous solution is concentrated, filtered, washed with methylene chloride, and then the aqueous solution is extracted. Dry over anhydrous sodium sulfate. The solvent was distilled off to obtain 0.20 g (yield 49%) of (R, R, R) -N- (picolinoyl) spiro [4,4] nonane-1,6-diamine (B).
IR (cm ^-1 ): 3361, 2959, 2874, 1664, 1591, 1568, 1529, 1466, 1435, 1202, 1174, 1130
[Example 2] Synthesis of compound (C) In the same manner as in Example 1, (S, S, S) -cis, cis-spiro [4,4] nonane-1,6-diamine ((S) -F), Hereinafter, the S-form compound is represented as (S)-. ) 1.00 g gave 0.58 g (yield 35%) of N- (t-butoxycarbonyl) diamine ((S) -F). (S, S, S) -N- (t-butoxycarbonyl) spiro [4,4] nonane-1,6-diamine ((S) -G) was used in the same manner as in Example 1 (S, S , S) -N- (t-butoxycarbonyl) -N`- (2-picolinoyl) spiro [4,4] nonane-1,6-diamine ((S) -H) 0.60 g (yield 73%) Obtained.

Performed from 0.20 g of (S, S, S)-(N- (t-butoxycarbonyl) -N`- (2-picolinoyl) spiro [4,4] nonane-1,6-diamine ((S) -H) In the same manner as in Example 1, 0.14 g (yield 97%) of (S, S, S) -N- (2-picolinoyl) spiro [4,4] nonane-1,6-diamine (C) was obtained.
IR (cm ^-1 ): 3364, 2957, 2870, 1666, 1591, 1568, 1520, 1467, 1435, 1199, 1171, 1128
Example 3 Synthesis of Compound (D) (R, R, R) Form (X = Cl) Potassium tetrachloroaurate (III) (KAuCl ₄ ) (0.61 g, 1.63 mmol) is added and dissolved in 15 ml of water. Next, a solution of 0.45 g (1.74 mmol) of (R, R, R) -N- (2-picolinoyl) spiro [4,4] nonane-1,6-diamine (B) dissolved in 60 ml of methanol was added and heated for 5 hours. Reflux. Filter through Celite, concentrate, add 15 ml of acetolyl and leave to crystallize. Cool and filter the precipitated crystals. The filtrate is washed with tolyl in aceto and dried. 0.43 g of gold complex was obtained. The results of instrumental analysis are as follows.

IR (KBr) cm ^-1 : 1685, 1605, 1505, 1466, 1387, 1348, 829, 756, 663
¹ H-NMR: 7.2-9.2 ppm (4H), 7.40 ppm (1H), 8.25 ppm (1H), 1.3-4.0 ppm (10H)
¹³ C-NMR: 170.4 ppm (C = O), 127.5-149.5 ppm (aromatic 4C), 57.2 ppm, 57.3 ppm (2C-N), 57.6 ppm (tC), 20.1-38.4 ppm (aliphatic 6C)
ESI-MS: The numerical value around m / z 490 showed that there was one Cl atom.
_{[C 15 H 20 N 3 ClOAu} ] matches ^+.

From the above results, it was confirmed that the compound of Example 3 had a chemical structure represented by the compound (D) (X = Cl).
Example 4 Synthesis of Compound (E) (S, S, S) Form (X = Cl) In the same manner as in Example 3, (S, S, S) -N- (2-picolinoyl) spiro [4 , 4] nonane-1,6-diamine (C) 0.38 g (1.46 mmol) gave 0.40 g of gold complex (E).
IR (KBr) cm ^-1 : 1654, 1607, 1487, 1447,, 1370, 1339, 797, 758, 667
As a result of IR measurement, it was confirmed that the compound of Example 4 had a chemical structure represented by the compound (E) (X = Cl) because it coincided with the complex (D).
[Example 5] Drug effect test A test solution was prepared by dissolving compound (D) in DMSO (dimethyl sulfoxide) at a concentration of 8 mg / ml.

The test consists of 5 types of human lung cancer cells (LU65A, LU99, A549, RERF-LC-MA, H460), 3 types of human gastric cancer cells (LKATO III, MKN-1, MKN-45), and human liver. 2 types of cancer cells (HuH-7, HepG2), 2 types of human pancreatic cancer cells (KP-1N, AsPC-1), 1 type of human colorectal cancer (DLD-1)
), 2 human colon cancer cells (HCT116, HT-29), 1 human rectal cancer cell (CaR-1), 3 human prostate cancer cells (PC-3, DU145, LNCap.FGC), human 1 skin cancer cell (A431), 1 human malignant melanoma cell (G-361), 1 human mesothelioma cell (H2452), 2 human esophageal cancer cells (TT, TE-6), human 1 type of bladder cancer cell (T24), 1 type of human cervical cancer cell (ME-180), 2 types of human lymphoma cells (U937, Jurkat E6.1), 1 type of human leukemia cells (HL60) It was.

These cells were suspended in each culture medium supplemented with 10% serum and dispensed into 96-well plates. Thereafter, the cells were cultured overnight at 37 ° C. in 5% CO ₂ . Test solutions were prepared in various concentrations in a culture medium and dispensed onto plates that had been pre-seeded with cells. The cells were further cultured for 3 days at 37 ° C. in 5% CO ₂ .

  Cell proliferation after drug addition was measured by MTS method (Promega's Cell Titer 96 Aqueous One Solution Cell Proliferation Assay) 1 to 3 days after drug addition.

  From the measured MTS value, the inhibition rate (%) of cell proliferation was determined by the following formula.

Inhibition rate (%) = (1-MTS value in the drug-added group / MTS value in the drug-free group) × 100
Since the value obtained by the above formula represents the cell growth inhibition rate, the higher the numerical value, the higher the drug effect. Those having a value of 50 (%) or more were regarded as having a drug effect. The results are shown below.

  From Table 1 above, human lung cancer cells (LU65A, LU99, A549, RERF-LC-MA), human gastric cancer cells (MKN-1), human liver cancer cells (HuH-7), human colon cancer cells (DLD) -1), human rectal cancer cells (CaR-1), human malignant melanoma cells (G-361), and human bladder cancer cells (T24) compared to oxaliplatin, a conventional anticancer agent A strong medicinal effect was observed.

  Further, the X axis of the graph shown in FIG. 1 represents the cell growth inhibition rate (%) obtained from the formula, and the Y axis represents the drug concentration (μg / ml). From FIG. 1, it was determined that normal hepatocytes (THLE-2) had low toxicity at a low concentration (less than 10 μg / ml) compared to oxaliplatin.

  As described above, the gold complex of the present invention has strong antitumor activity and low toxicity, and is useful as a malignant tumor therapeutic agent.

Claims (8)

A gold complex having N- (2-picolinoyl) spiro [4,4] nonane-1,6-diamine represented by the following three-dimensional structural formula (A) as a ligand.

In Claim 1, it has (R, R, R) -N- (2-picolinoyl) spiro [4,4] nonane-1,6-diamine represented by the following three-dimensional structural formula (B) as a ligand. Gold complex.

In Claim 1, it has (S, S, S) -N- (2-picolinoyl) spiro [4,4] nonane-1,6-diamine represented by the following three-dimensional structural formula (C) as a ligand. Gold complex.

3. The gold complex according to claim 2, wherein the gold complex is a gold complex represented by the formula (D).

(Here, X represents a halogen or a monovalent anion.) 4. The gold complex according to claim 3, wherein the gold complex is a gold complex represented by the formula (E).

(Here, X represents a halogen or a monovalent anion.)   6. The gold complex according to claim 4, wherein the gold complex is X is chlorine.   A pharmaceutical composition comprising the gold complex according to claim 1 as an active ingredient.   A therapeutic agent for malignant tumors comprising the gold complex according to claim 1 as an active ingredient.

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