KR101593756B1 - Novel tetranuclear arene-Ru based metalla-bowls compound and pharmaceutical composition for preventing or treating cancer containing the same as active ingredient - Google Patents

Abstract

The present invention relates to a novel 4-nuclear arene-ruthenium-based bowel compound or a pharmaceutically acceptable salt thereof, which is useful for the treatment of human HCT-15 (rectal cancer), SK-hep-1 (liver cancer) and AGS Inhibits proliferation at a low micromolar concentration, exhibits excellent anticancer activity by inhibiting the activity of cancer cells by increasing the expression of a tumor suppressor, and thus can be effectively used for prevention and treatment of cancer.

Description

TECHNICAL FIELD [0001] The present invention relates to a novel tetranuclear arene-ruthenium-based bowl-shaped compound and a pharmaceutical composition for preventing or treating cancer containing the same as an active ingredient. ingredient}

The present invention relates to a novel 4-nuclear arene-ruthenium-based bowl-shaped compound or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition for preventing or treating cancer containing the same as an active ingredient.

The field of metal pharmacy has emerged as an important new area of medicinal chemistry due to the therapeutic application of metal based drugs ((a) M. Mascini, G. Bagni, MLD Pietro, M. Ravera, S. Baracco and D. Osella, , 2006, 19, 409; (b) TW Hambley, Dalton Trans., 2007, 4929). The rearrangement of the organic ligands in a three-dimensional space from a wide range of coordination numbers and the accessible oxidation-reduction state of the adjustable metal centers provide a broad spectrum of reactivity that can be used for medicinal purposes (U. Schatzschneider and N. Metzler- Nolte, Angew. Chem., Int. Ed., 2006, 45, 1504.). Platinum complexes in particular cisplatin, carboplatin, and oxoplatin have been shown to be effective in treating cancer patients despite their high toxicity and unnecessary nerve, liver, and renal toxic side effects ((a) Y. Jung and SJ Lippard, Chem. Rev., (B) C. Gianomenico and MUS Christen, Patent 6413953, 2000), are currently being used as the most effective chemical therapeutic agents ((a) L. Kelland, Nat. Rev. Cancer, 2007, Reedijk, Eur., J. Inorg. Chem., 2009, 1303). However, the high systemic toxicity and resistance problems associated with platinum-based drugs have been addressed by alternative metal antineoplastic agents (CHA Goss, W. Henderson, AL Wilkins and CJ Evans, J. Organomet. Chem. 2003, 679, (C) W. Henderson, BK Nicholson and ERT Tiekink, Inorg. Chim. Acta, 2006, 359, 2046) and safer and more effective Opened a new era of interest in the design and pharmacological development of therapeutic agents. In particular, the ruthenium complex represents a new class of promising metal-based drugs with low toxicity and high activity in tumors that do not respond well to platinum-based drugs ((a) CG Hartinger, S. Zorbas-Selfried, MA Jakupee, B. 2006, 100, 891; (b) YK Yan, M. Melchart, A. Habtemariam and PJ Sadler Chem. Commun., 2005, 4764). Two ruthenium complexes, ImH [Trans RuCl4 (DMSO) Im]] (NAMI-A) and KO1019 were the first nuclear ruthenium anticancer drugs to successfully pass the Phase 1 clinical trial ((a) JM Rademaker-Lakhai, (B) M. Groessl, CG Hartinger, K. Polec-Pawlak, M. Jarosz, PJ Dyson and BK Keppler (2004), Dennis Bongard, D. Pluim, JH Beijnen and JHM Schellens, Clin. Cancer Res., 2004, , Chem. Biodiversity, 2008, 5, 1609.).

Polynuclear drugs have also been designed to increase the range of treatable tumors. Many polymer platinum compounds ((a) Z. Yang, X. Wang, H. Diao, J. Zhang, H. Li, H. Sung and Z. Guo, Chem. Commun., 2007, , AI Day, RJ Blanch, AP Arnold, C. Cullinane and JG Collins, Chem. Commun., 2004, 1424), for example polymer linked diaminocyclohexyl metal chemically curable (AP5346) and pt- A polyglycerol polymer with branched structure is formed through selective cross-linking (EPR) (Y. Matsumura and H. Maeda, Cancer Res., 1986, 46, 6387.) and long range strand liver and strand DNA cross-linking May be potentially useful for target-specific tumor cells because of their unique extracellular environment that allows the drug to selectively accumulate in cancer cells. Accordingly, studies on the polynuclear metal-polymer complex compounds have been actively conducted.

Korea Public Patent 2012-0101798

The present invention provides a novel 4-nuclear arene-ruthenium-based bowl-shaped compound which is a kind of polynuclear metal-polymer complex and its use for chemotherapy.

In order to solve the above-mentioned object, the present invention provides a quaternary arene-ruthenium-based bowl-shaped compound represented by the following formula 1 or a pharmaceutically acceptable salt thereof.

≪ Formula 1 >

는 하기 화학식 2 내지 4로 표시되는 화합물 중 하나임).(In the formula 1,

Is one of the compounds represented by the following formulas (2) to (4)).

(2)

(3)

≪ Formula 4 >

The present invention also provides a pharmaceutical composition for the prevention or treatment of cancer comprising the above-mentioned novel 4-nuclear arene-ruthenium-based bowl-shaped compound or a pharmaceutically acceptable salt thereof as an active ingredient.

The quaternary arene-ruthenium-based bowl-shaped compound according to the present invention inhibits the proliferation of human HCT-15 (rectal cancer), SK-hep-1 (liver cancer) and AGS (gastric cancer) cells at low micromolar concentrations, And exhibits excellent anticancer activity by inhibiting the activity of cancer cells by increasing expression, and thus can be usefully used for prevention and treatment of cancer.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a ¹ H NMR spectrum of a compound of Example 1 which is a bowl-shaped compound according to an embodiment of the present invention. FIG.
2 is a ¹ H NMR spectrum of the compound of Example 2, which is a bowl-shaped compound according to an embodiment of the present invention.
3 is a ¹ H NMR spectrum of the compound of Example 3, which is a bowl-shaped compound according to an embodiment of the present invention.
4 is a ¹³ C NMR spectrum of the compound of Example 1, which is a bowl-shaped compound according to an embodiment of the present invention.
5 is a ¹³ C NMR spectrum of the compound of Example 2, which is a bowl-shaped compound according to an embodiment of the present invention.
6 is a ¹³ C NMR spectrum of the compound of Example 3, which is a bowl-shaped compound according to an embodiment of the present invention.
7 is a graph of a theoretical (top) and experimental (bottom) ESI-MS result for self-assembly [2 + 2] of Bowl Compounds Examples 1 to 3 according to one embodiment of the present invention.
8 (a) is an X-ray crystal structure of a bowl-shaped compound of Example 1 according to an embodiment of the present invention (solvent molecules, counter anions, and hydrogen atoms are omitted for the sake of clarity), (b) Is a space-filling CPK model of a bowl-shaped compound of Example 1 according to one embodiment of the present invention.
9 is a methanol (1 × 10 ^-5 M) in donor 1, acceptor 2 - 4 and shows the electronic absorption spectrum of the bowl-shaped compound, according to one embodiment of the present invention.
Figure 10 is a graph showing the results of a < RTI ID = 0.0 > Lt; RTI ID = 0.0 > RT-PCR < / RTI > analysis of APC expression in HCT116 cells.
Figure 11 is a graph showing the effect of the compound of Example 2 treated with the compound of Example 2 Lt; RTI ID = 0.0 > RT-PCR < / RTI > analysis of p53 expression in HCT116 cells.
12 is a graph showing the loss of growth inhibitory activity of the compound of Example 2 according to an embodiment of the present invention, depending on the solvent for HCT-15 cells.

The present invention provides a quaternary arene-ruthenium-based bowl-shaped compound represented by the following general formula (1) or a pharmaceutically acceptable salt thereof:

≪ Formula 1 >

는 하기 화학식 2 내지 4로 표시되는 화합물 중 하나임).(In the formula 1,

Is one of the compounds represented by the following formulas (2) to (4)).

(2)

(3)

≪ Formula 4 >

The quaternary arene-ruthenium-based bowl-like compound of the present invention represented by the above-mentioned formula (1) can be used in the form of a pharmaceutically acceptable salt, and the salt includes the acid addition salt formed by a pharmaceutically acceptable free acid Is useful. As the free acid, inorganic acid and organic acid can be used. As the inorganic acid, hydrochloric acid, bromic acid, sulfuric acid, sulfurous acid, phosphoric acid and the like can be used. As the organic acid, citric acid, acetic acid, maleic acid, fumaric acid, , Acetic acid, glycolic acid, succinic acid, tartaric acid, 4-toluenesulfonic acid, galacturonic acid, embonic acid, glutamic acid, citric acid and arpartic acid. Preferably, hydrochloric acid is used as the inorganic acid, and methanesulfonic acid is used as the organic acid.

In addition, the quaternary arene-ruthenium-based bowl-like compound represented by the above formula (1) of the present invention includes not only pharmaceutically acceptable salts, but also all salts, hydrates and solvates which can be prepared by a conventional method .

The addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the compound of Chemical Formula 1 in a water-miscible organic solvent such as acetone, methanol, ethanol, acetonitrile, etc., And then precipitating or crystallizing the acid solution. Subsequently, in this mixture, a solvent or an excess acid is evaporated and dried to obtain an additional salt, or the precipitated salt may be produced by suction filtration.

The quaternary arene-ruthenium-based bowl-shaped compound of Formula 1 according to the present invention is a compound of Formula 2, which is a N, N'-bis (4- (pyridin-4-ylethynyl) phenyl) terephthalamide donor ligand, -Ruthenium acceptor in a ratio of 1: 1, and the reaction can be performed in a 1: 1 solution of nitromethane-dichloromethane, but is not limited thereto. The reaction can take place under stirring at room temperature for 1 to 10 hours, and the addition of diethyl ether to the concentrated reaction mixture results in the formation of a pure self-assembled product of formula (1).

As described above, the novel 4-nuclear arene-ruthenium-based bowl-shaped compound prepared according to the present invention can be produced by a method such as infrared spectroscopy, nuclear magnetic resonance spectrum, mass spectrometry, liquid chromatography, X- Ultraviolet / visible light absorption spectroscopy and elemental analysis of representative compounds The molecular structure can be identified by comparison of calculated and measured values.

As shown in the following examples, the 4-nuclear arene-ruthenium-based bowl-shaped compound of formula (1) according to the present invention can be used for the treatment of human SK-hep-1 (liver cancer), AGS (gastric cancer) and HCT- Inhibits proliferation at low micromolar concentrations, inhibits the activity of cancer cells through apoptosis that regulates the cell growth cycle, and exhibits excellent anticancer activity by increasing the expression of tumor suppressors such as APC and p53. It can be used as an active ingredient. Accordingly, the present invention relates to a pharmaceutical composition for preventing or treating cancer comprising the above-mentioned 4-nuclear areene-ruthenium-based bowl-shaped compound or a pharmaceutically acceptable salt thereof as an active ingredient, the 4-nuclear areen- A method for treating cancer comprising administering to a subject a 4-nuclear arene-ruthenium-based bowl-shaped compound.

In one embodiment of the present invention, the pharmaceutical composition for preventing or treating cancers containing the 4-nuclear areene-ruthenium-based bowl-shaped compound or a pharmaceutically acceptable salt thereof as an active ingredient is a 4 The present invention can include 0.01 to 90 parts by weight, 0.1 to 90 parts by weight, 1 to 90 parts by weight, or 10 to 90 parts by weight of a nuclear areene-ruthenium-based bowl compound or a pharmaceutically acceptable salt thereof, but is not limited thereto , The condition of the patient, the type of disease and the degree of progression.

In another embodiment of the present invention, the pharmaceutical composition for preventing or treating cancers containing the 4-nuclear Areen-ruthenium-based bowl-shaped compound or a pharmaceutically acceptable salt thereof as an active ingredient is a carrier, an excipient, a disintegrant, The composition may further comprise at least one auxiliary agent selected from the group consisting of coating agents, swelling agents, lubricants, lubricants, flavors, antioxidants, buffers, bacteriostats, diluents, dispersants, surfactants, binders and lubricants.

Specific examples of carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. Solid formulations for oral administration may be in the form of tablets, pills, powders, granules, capsules These solid preparations can be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., into the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, syrups and the like, and various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin which are commonly used simple diluents. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As the suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

In another embodiment of the present invention, the formulation of the pharmaceutical composition for preventing or treating cancer comprising the 4-nuclear Areen-ruthenium-based bowl-shaped compound or a pharmaceutically acceptable salt thereof as an active ingredient is a granule, a powder, , Tablets, pills, capsules, suppositories, gels, syrups, juices, suspensions, emulsions, drops or solutions.

According to one embodiment of the present invention, the pharmaceutical composition may be administered orally, intraarterally, intraperitoneally, intramuscularly, intraarterally, intraperitoneally, intrasternally, transdermally, nasally, inhaled, topically, rectally, ≪ / RTI > can be administered to the subject in a conventional manner.

The preferred dose of the pharmaceutical composition for preventing or treating cancer comprising the 4-nuclear islet-ruthenium-based bowl-shaped compound or a pharmaceutically acceptable salt thereof as an active ingredient depends on the condition and body weight of the patient, Type, route of administration, and duration of administration, and may be appropriately selected by those skilled in the art. According to one embodiment of the present invention, the daily dose may be 0.01 to 1,000 mg / kg, specifically 0.1 to 1,000 mg / kg, more specifically 0.1 to 100 mg / kg, though it is not limited thereto. The administration may be performed once a day or divided into several times, and thus the scope of the present invention is not limited thereto.

In the present invention, the 'subject' may be a mammal including a human, but is not limited thereto.

In one embodiment of the present invention, the cancer may be solid cancer, and more specifically, the 4-nuclear arene-ruthenium-based bowl-like compound of Formula 1 according to the present invention may be used as a brain tumor, a benign astrocytoma ), Malignant astrocytoma, pituitary adenoma, meningioma, CNS lymphoma, oligodendroglioma, craniopharyngioma, ependymoma, brain tumors Brainstem tumor, Head & Neck tumor, Larygeal cancer, Oropgaryngeal cancer, Nasal cavity / PNS tumor, Nasopharyngeal tumor, Salivary gland tumor, small cell lung cancer, non-small cell lung cancer, hypopharyngeal cancer, thyroid cancer, oral cavity tumor, Chest Tumor, , Thymoma Esophageal cancer, Breast cancer, Male breast cancer, Abdomen-pelvis tumor, Stomach cancer, Hepatoma, Gallbladder cancer, Esophageal cancer, Breast cancer, Small bowel cancer, gall bladder cancer, billiary tract tumor, pancreatic cancer, small intestinal tumor, large intestinal tumor, anal cancer, bladder cancer, ), Renal cell carcinoma, male genital cancer, penile cancer, prostate cancer, female genital cancer, cervix cancer, Endometrial cancer, ovarian cancer, uterine sarcoma, vaginal cancer, female vulvar cancer, female urethral cancer or skin cancer) cancer. < / RTI > And more preferably for the treatment of liver cancer, stomach cancer or rectal cancer.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples in order to facilitate understanding of the present invention. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

1. General

The reactants used in this experiment were purchased commercially and used without further purification. ^{The 1} H and ¹³ C NMR spectra were recorded on a Bruker 300 MHz spectrometer. The chemical shift (?) In the ¹ H NMR spectrum was recorded as relative ppm values with tetramethylsilane (Me ₄ Si) as the international standard (0.0 ppm). The mass spectra were recorded on a Micromass Quattro II triple-quadrupole mass spectrometer using an electron spray ionization with a MassLynx operating system. Elemental analyzes were performed using an Elemental GmbH Vario EL-3 instrument. The UV-Vis spectrum was recorded on the Cary 100 Conc. Fluorescence titration studies were performed with the HORIBA FluoroMax-4 fluorescence intensity meter.

< Example  1-3> New  4 nuclear Areen - Manufacture of ruthenium-based bowl-shaped compounds

In an embodiment of the present invention, as shown in Reaction Scheme 1 below, AgO₃SCF₃In the presence of 2-nuclear ruthenium ruthenium complex [Ru₂- (Allen)₂ (OO ∩OO) · Cl₂] (2, OO ∩OO = 2,5-dioxido-1,4-benzoquinone;3, OO ∩OO = 5,8-dioxido-1,4-naphthacquinone;4, OO ∩OO = lauric acid) and bis-pyridine ligandsOneof 1: 1 molar ratio to prepare a 4-nuclear arene-ruthenium-based bowl-shaped compound.

Specifically, one of the compounds 2 to 4 (0.1 mmol) and 2 equivalents of AgCF ₃ SO ₃ (0.2 mmol) were added to methanol, stirred at room temperature for 2 hours, and then filtered to remove AgCl. Compound 1 (0.1 mmol), the donor ligand corresponding to the filtrate, was then added. The mixture was stirred at room temperature for 12 hours and then the solvent was removed under reduced pressure. The residue was taken up in a nitromethane / methanol (1: 1) solution and the extract was filtered and concentrated. The product was precipitated by adding diethyl ether to give Example 1 (Compound 5 ) or Example 2 (Compound 6 ) Or the compound of Example 3 (Compound 7 ) was collected.

<Reaction Scheme 1>

The compound of Example 1:

Red crystalline solid of 90% yield.

Elemental analysis of C ₉₀ H ₈₆ F ₁₂ N ₁₀ O ₂₄ Ru ₄ S ₄ Calculated: C, 44.08; H, 3.53; N, 5.71; Measured: C, 44.18; H, 3.70; N, 5.90.

MS (ESI): calculated for [M ² OTf] ²⁺ m / z 1077.59.6, found 1077.32; Calculated for [M 3 OTf] ³⁺ m / z 668.74, found 668.70.

Absorption spectrum [λ _max , CH ₃ OH]: 305 and 500 nm.

¹ H NMR (nitromethane-d ₃ , 300 MHz,?, Ppm): 10.61 (s, 4H, CONH), 8.16-8.10 (m, 12H, H _a / H _c ), 8.05-7.92 _{H d / H b), 6.05} (m, 8H, H cym), 5.91-5.75 (m, 12H, H cym / H benz), 2.95-2.87 (m, 4H, -CH (CH 3) 2), 2.19 _{(s, 12H, -CH 3)} , 1.36 (d, 24H, -CH (CH 3) 2).

¹³ C NMR (nitromethane _{-d 3, 300 MHz, δ,} ppm): 185.14 (C = O), 185.07 163.83, 153.89, 148.90, 148.56, 141.66, 127.81, 124.29, 120.40, 120.65, 117.44, 105.43, 102.87, 102.40, 100.29, 84.81, 84.63, 82.81, 82.73, 32.67, 22.59, 22.57, 18.45.

The compound of Example 2:

91% yield of a green crystalline solid.

C ₉₈ H ₉₀ F ₁₂ N ₁₀ O ₂₄ Elemental analysis for Ru ₄ S ₄ Calculated: C, 46.12; H, 3.55; N, 5.49; Measured: C, 46.40; H, 3.72; N, 5.35.

MS (ESI): calculated for [M ² OTf] ²⁺ m / z 1127.60, found 1127.82; Calculated for [M 3 OTf] ³⁺ m / z 702.09, found 702.07.

Absorption spectrum _{[λ max, CH 3 OH]} : 290 and 450nm.

¹ H NMR (nitromethane _{-d 3, 300 MHz, δ,} ppm): 10.42 (s, 4H, CONH), 8.25-8.17 (m, 12H, H a / H c), 8.10 (t, 2H, H d ), 7.82 (d, 8H, J = 6.0 Hz, H b), 7.24 (d, 8H, J = 6.0 Hz, H benz), 5.77 (d, 8H, J = 6.0 Hz, H cym), 5.56 (d , 8H, J = 6.0 Hz, H cym), 2.95-2.86 (m, 4H, -CH (CH 3) 2), 2.17 (s, 12H, -CH 3), 1.38 (d, 24H, -CH (CH ₃ ) ₂ ).

¹³ C NMR (nitromethane _{-d 3, 300 MHz, δ,} ppm): 172.58 (C = O), 172.16 163.65, 153.32, 148.79, 148.29, 138.96, 138.68, 124.29, 127.18, 116.86, 112.79, 105.10, 100.40, 85.25, 84.48, 32.02, 22.50, 22.50, 17.49.

Compound of Example 3:

Yellow crystalline solid of 90% yield.

C ₈₄ H ₈₄ F ₁₂ N ₁₆ O ₂₄ Elemental analysis of Ru ₄ S ₄ Calculated: C, 40.98; H, 3.44; N, 9.10; Measured: C, 40.80; H, 3.62; N, 9.35.

MS (ESI): calculated for [M 2OTf] ²⁺ m / z 1094.59, found 1094.32; Calculated for [M 3 OTf] ³⁺ m / z 680.08, found 680.00.

Absorption spectrum _{[λ max, CH 3 OH]} : 292 and 400nm.

¹ H NMR (nitromethane _{-d 3, 300 MHz, δ,} ppm): 11.75 (s, 4H, CONH, ligand), 10.57 (s, 2H, CONHCO, receptors), 8.14-8.11 (m, 12H, H a _{/ H c), 7.98-7.90 (m} , 10H, H d / H b), 6.20-6.13 (m, 8H, H cym), 5.86-5.78 (m, 8H, H cym), 2.86-2.78 (m, _{4H, -CH (CH 3) 2} ), 1.95-1.87 (m, 12H, -CH 3), 1.34-1.21 (m, 24H, -CH (CH 3) 2).

¹³ C NMR (nitromethane _{-d 3, 300 MHz, δ,} ppm): 179.91 (C = O), 163.55 (C = O), 155.90, 153.87, 148.75, 141.52, 128.04, 127.79, 124.26, 120.02, 106.98, 101.11, 86.01, 85.49, 84.47, 81.55, 80.84, 32.45, 22.82, 22.75, 22.71, 22.61, 22.51, 18.25.

&Lt; Experimental Example 1 >

One. NMR  analysis

^{The 1} H and ¹³ C NMR spectra were recorded on a Bruker 300 MHz spectrometer.

The chemical shift (?) In the ¹ H NMR spectrum was recorded as relative ppm values with tetramethylsilane (Me ₄ Si) as the international standard (0.0 ppm).

The ¹ H NMR spectra of the compounds of Examples 1 to ₃ in nitromethane-d ₃ are shown in FIGS. 1 to 3, respectively, and ¹³ C NMR spectra are shown in FIGS. 4 to 6, respectively.

As shown in FIGS. 1 to 3, the compounds of Examples 1 to 3 according to the present invention clearly showed the formation of symmetrical species with typical resonance movements occurring during metal complex formation. The signal corresponding to the H _a nucleus on the pyridine ring shifted to the upfield (0.1-0.6 ppm) compared to ligand 1, presumably due to current shielding, which is due to the downfield ).

Similarly, as shown in Figs. 4 to 6, the 13 C NMR spectra of the compounds of Examples 1 to 3 according to the present invention are very clearly shown and are well decomposed.

2. Mass spectrum analysis

High resolution electron spray ionization mass spectrometry (HR-ESI-MS) data was also obtained.

The mass spectra were recorded on a Micromass Quattro II triplequadrupole mass spectrometer using electrospray ionization with a MassLynx operating system. The results are shown in Fig.

Examples of the charge state of the compound 1 was observed at m / z = 1077.32 ([M -2OTf] 2+) and ^{668.70 ([M-3OTf] 3+} ) ( corresponding to the top of FIG. 7, Compound 5), and the charged state of the embodiment of example 2, the compound was observed at m / z = 1127.82 ([m -2OTf] 2+) and ^{702.07 ([m-3OTf] 3+} ) ( corresponding to the intermediate of Fig. 7, compound 6), carried out The charge state of the compound of Example 3 was observed at m / z = 1094.32 ([M-2OTf] ²⁺ ) and 680.00 ([M-3OTf] ³⁺ ) (corresponding to compound 7 at the bottom of FIG. These peaks (red graph) correspond to the theoretical distribution (blue graph), which strongly supports the formation of self-assembled products [2 + 2].

3. Single Crystal X-ray diffraction  analysis

The molecular structure of the compound of Example 1 was clearly identified by single crystal X-ray diffraction analysis

Crystals suitable for structural analysis were grown by slow diffusion of diethyl ether into a solution of the compound of Example 1 dissolved in a mixed solvent of CH ₃ OH / CH ₃ NO ₂ (1: 1).

The crystals were coated with paratone oil and diffraction data were measured with Mo K? Radiation at 173 K on an X-ray diffraction camera system using an imaging plate equipped with a graphite crystal incoming linear monochromator. RapidAuto software was used for data collection and data processing. The crystal structure was identified by direct methods using the SIR2008 software package and refined by full-matrix least-squares calculation using the SHELXTL software package. Final scouring was carried out with a modification of the structural parameters using the SQUEEZE option in the PLATON. This procedure is used to measure three methanol molecules with large displacement parameters, the electron density of the four water molecules, and the electron density of the solvent region, which is not completely regular. All non-hydrogen atoms are anisotropically refined; The hydrogen atoms were assigned to the isotropic displacement coefficients U (H) = 1.2 U (C) and 1.5 U (C _methyl ), and their coordinates depend on their respective atomic coordinates. Least-squares refinement of the structural model can be performed under geometric constraints such as DFIX, DANG, FLAT, SAME, EADP, and EXYZ, and in p-cymene, triflate, diethyl ether, and methanol Were performed under the constraints of displacement parameters such as ISOR, DELU and SIMU. The refinement of the structure yields the final R1 = 0.1105 and wR2 = 0.3068 for 21388 reflections with l> 2σ (l); Converged at R1 = 0.1903 and wR2 = 0.3592 for 6897 reflections. The greatest different peaks and holes were 1.093 and -0.992 e Å ³ , respectively.

The results are shown in Fig.

Figure 8 (a) shows the X-ray crystal structure of the compound of Example 1, wherein the four Ru2 + centers and the bowl-shaped bispyridine ligands are shown. 8 (b) shows a space-filling CPK model of the compound of Example 1. Fig. Solvent molecules, counter anions, and hydrogen atoms have been omitted for clarity.

8, the resulting structure is shown in _{[2 + 2] M 4 L} 2 two [Ru ₂ (arene) forming a bowl-shaped metal compound ₂ (5,8-di-oxido-1,4- Naphthacquinoneato) ₂ ] ⁴⁺ building block. Each cement-coated Ru coordination sphere contains nitrogen atoms and two oxygen atoms defining metal-bowl-shaped dipyridyl and naphthacenedioneates peaks, respectively, and they have four nuclear frames with internal dimensions of 8.3 x 13.31 &Lt; / RTI &gt; Bond length of _N-pyr Ru1 and Ru2-N _pyr while each of 2.13 (2) 2.12 and (2), and _pyr Ru3-N-N _pyr Ru4 each length was 2.11 (2) 2.12 and (2). The angle of 82.5 DEG to 87.1 DEG of the NRuO slice slightly deviated from the ideal 90 DEG.

4. Electronic absorption spectrum

The electronic absorption spectra of the compounds of Examples 1 to 3 (together with their corresponding metal acceptors (compounds of formulas 2 to 4)) were observed in a 1 × 10 ^-5 M methanol solution and are shown in FIG.

9, the absorption spectrum in Example λ _abs = with respect to the compound of 1 305 and 500 nm, examples of a compound represented by the 2 λ _abs = 290 and 450 nm, and carried out on the compound of Example 3 λ _abs = = 292 and 400 nm. Although all complexes show considerable migration after donor ligand 1 and coordination, the strong band of the compounds of Examples 1-3 exhibit similar characteristics to the starting receptor 2-4 . The high energy band observed in all example compounds was also within the spectrum of the free ligand 1. 2 nuclear arene-Ru receptor also exhibited high-energy absorption at 290330 nm, as well as broad and low-energy absorption at 400500 nm. As with the use of pyridyl donor bands, these arene-Ru bands are also conserved for self-assembly, which results in strong absorption for the example compounds. These bands appear to be a combination of charge transfer transitions from metal-to-ligand to intermolecular / intermolecular π → π ^* transition.

<Experimental Example 2> Cancer cell growth inhibition assay

For the analysis of the biological effects of the compounds of the present invention, donor 1 (II), 2 to 4 receptor Human HCT-15 of the compound of (formula 3) to (5) and the compound of Examples 1 to 3 (cancer), SK-hep- 1 (liver cancer) and AGS (gastric cancer) cell line.

More specifically, cancer cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 1% penicillin streptomycin and RPMI 1640 at 37 ° C and 5% CO ₂ . The suspension of each of the other cells was dispensed into 96 well plates at a concentration of 1 x 10 ⁴ cells / well. After incubation for 24 hours, the cells were treated with donor 1 (Formula 2), Receptor 2-4 (Formula 3 to 5) compounds and Compounds of Examples 1 to 3 and cisplatin or doxorubicin ) Were treated with 1, 6, 8, 40 and 200 μM. Next, MTT ((3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) was dissolved in phosphate buffer (PBS, pH 7.2) and filtered with a 0.22 μm millipore filter After incubation for 3 hours at 37 ° C and 5% CO ₂ , the MTT solution was removed and the cells were lysed for cytolysis 100 μL of DMSO (dimethylsulfoxide) was added to each well. The plates were then read in absorbance at 550 nm with a multi-reader (Tecan, Switzerland) to determine the ratio of the absorbance The IC ₅₀ values for cell growth inhibition were determined by fitting a plot of the log percentage of surviving cells to the log of drug concentration using a linear regression function ).

Cytotoxicity of donor 1 , receptor 2-4 compounds and compounds of Examples 1 to 3 is shown in Table 1.

compound IC ₅₀ (uM) HCT-15 SK-hep-1 AGS Donor 1 132.7 ± 6.85 120.4 + 1.66 70.1 ± 6.57 Receptor 2 136.8 ± 1.02 84.5 ± 2.24 63.3 ± 2.90 Receptor 3 146.0 ± 9.88 87.0 ± 10.39 30.6 ± 0.78 Receptor 4 38.2 ± 2.74 34.0 + 1.36 6.9 ± 0.46 Example 1 > 200 119.2 ± 3.54 23.0 + - 0.72 Example 2 6.9 ± 0.13 4.3 ± 0.15 4.6 ± 0.10 Example 3 > 200 95.9 ± 2.65 31.7 ± 0.83 Cisplatin 13.2 ± 0.84 14.9 ± 1.08 102.8 ± 3.61 Doxorubicin 15.9 ± 0.92 6.4 ± 0.13 5.2 ± 0.12 IC ₅₀ : drug concentration required for 50% inhibition of cell viability; Data are expressed as mean ± standard error (SE).

Referring to Table 1, it was found that all of the three cancer cell lines were sensitive to the compound of Example 2, and cell growth was effectively inhibited even with a very small amount of the compound treatment of Example 2. [ Particularly, in the case of the compound of Example 2, the IC ₅₀ value of 4-6 μM in the three cancer cell lines shows that the cancer cell is much smaller than the amount of cisplatin or doxorubicin used conventionally as an anticancer drug And thus the anticancer activity is exhibited. Therefore, it is considered that the compound of the present invention can be used as an excellent anticancer agent that can replace conventional cisplatin and doxorubicin.

Experimental Example 3 Quantitative RT-PCR Analysis

The growth of cancer cells is associated with cell cycle control and tumor suppressor. Thus, chemotherapeutic drugs currently used in hospitals include agents that target such regulatory proteins to inhibit tumor growth. In rectal cancer, genetic modification of APC and p53 is known to promote tumor growth. The APC gene plays an important role in cell cycle arrest and apoptosis, and the impairment of APC function is related to the development of rectal tumor formation. Known as tumor suppressors, p53 plays an important role in cellular responses to DNA damage, oxidative stress, and hypoxia, which also frequently changes in rectal cancer cells.

Thus, in this example, quantitative RT-PCR was performed to investigate the effect of the compound of Example 2 on the levels of APC and p53 mRNA in HCT116 colon cancer (wild type).

Total RNA was extracted from HCT116 rectal cancer cells (containing the wild-type p53 gene) using a PureLink ^™ RNA Mini Kit (Ambion, USA). 1 μg of the total RNA was reverse-transcribed in an amount of 20 μl using an oligo (dT) primer with the enzyme and buffer supplied to the PrimeScript II 1st strand cDNA synthesis kit (Takara, Japan). A quantitative real-time PCR reaction was performed on MX3005P (Stratagene, USA) using the following primers; APC was 5'-GGAGTAAAACTGCGGTC-3 ', 5'-GTACTTTCTCTGCTTCCATT-3', and p53 was 5'-CAGCCAAGTCTGTGACTTGCACGTAC-3 ', 5'-CTATGTCGAAAAGTGTTTCTGTCATC-3', β-actin was 5'-GTCCACCGCAAATGCTTCTA- 5'-TGCTGTCACCTTCACCGTTC-3 '. For real-time PCR, SYBR Premix Ex Taq II (Takara, Japan) was used. The final volume of the reaction was 25 μL, including 2 μL cDNA template, 12.5 μL Master Mix, 1 μL of each primer (10 μM stock solution), and 8.5 μL sterile distilled water. The thermal cycling profile consisted of a pre-incubation step at 95 ° C for 3 minutes followed by 40 cycles of 95 ° C (5 seconds) and 60 ° C (30 seconds). The relative quantitative evaluation of APC and p53 gene levels was performed by comparative CT (cycle threshold) method.

As a result, exposure of the compound of Example 2 at 2 [mu] M increased mRNA expression of APC (2.9-fold) as shown in Fig. In addition, as shown in Figure 11, p53 mRNA expression in HCT116 cells treated with 2 [mu] M Example 2 significantly increased (4.1-fold) compared to untreated control.

Therefore, the 4-nuclear arene-ruthenium-based bowl-shaped compound according to the present invention exhibits excellent anticancer activity by increasing the expression of tumor suppressors such as APC and p53 in cancer cells, and thus it can be used as a novel anticancer agent.

Experimental Example 4 Stability of the prepared compound

To measure the stability of the 4-nuclear arene-ruthenium-based bowl-shaped compound of the present invention, a 10 μM solution of the compound of Example 2 was pre-incubated at 37 ° C. for 0, 12, 24 and 48 hours in cell culture medium and DMSO , And then cell growth on HCT-15 rectal cancer cells was measured.

As shown in FIG. 12, in the growth inhibitory activity of the compound of Example 2, a 50% loss was observed in about 24 hours after the culture in the cell culture medium. On the other hand, in the presence of DMSO, its growth-inhibiting activity was stable until about 48 hours after the cultivation. These results suggest that the 4-nuclear arene-ruthenium-based bowl-shaped compound according to the present invention is stable in DMSO.

Meanwhile, the compound according to the present invention can be formulated into various forms depending on the purpose. The following examples illustrate some formulations containing the compound according to the present invention as an active ingredient, but the present invention is not limited thereto.

Formulation Example 1 Tablet (direct pressurization)

After 5.0 mg of the active ingredient was sieved, 14.1 mg of lactose, 0.8 mg of crospovidone USNF and 0.1 mg of magnesium stearate were mixed and pressed to prepare tablets.

&Lt; Formulation Example 2 > Tablets (wet assembly)

After 5.0 mg of the active ingredient was sieved, 16.0 mg of lactose and 4.0 mg of starch were mixed. 0.3 mg of Polysorbate 80 was dissolved in pure water, and an appropriate amount of this solution was added, followed by atomization. After drying, the granules were sieved and mixed with 2.7 mg of colloidal silicon dioxide and 2.0 mg of magnesium stearate. The fine particles were pressed to prepare tablets.

&Lt; Formulation Example 3 > Powder and capsule

5.0 mg of the active ingredient was sieved and then mixed with 14.8 mg of lactose, 10.0 mg of polyvinylpyrrolidone and 0.2 mg of magnesium stearate. The mixture was extruded through a hard No. 5 &lt; / RTI &gt; gelatin capsules.

&Lt; Formulation Example 4 >

100 mg of the active ingredient, 180 mg of mannitol, 26 mg of Na ₂ HPO _{4 12} H ₂ O and 2974 mg of distilled water were mixed. The mixed solution was filled in an ampoule made of transparent glass, sealed in an upper lattice by dissolving the glass, sterilized by autoclaving at 120 DEG C for 15 minutes or longer, and injected.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that such detail is solved by the person skilled in the art without departing from the scope of the invention. will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (8)

A quaternary arene-ruthenium-based bowl-shaped compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
&Lt; Formula 1 >

(1)

Is a compound represented by the following formula (3)
(3)

.
delete A pharmaceutical composition for preventing or treating cancer comprising the 4-nuclear arene-ruthenium-based bowl-shaped compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient. The method of claim 3,
Wherein the 4-nuclear arene-ruthenium-based bowl-shaped compound or a pharmaceutically acceptable salt thereof exhibits anticancer activity through apoptosis of cancer cells. The method of claim 3,
The pharmaceutical composition for the prevention or treatment of cancer may be a carrier, an excipient, a disintegrant, a sweetener, a coating agent, a swelling agent, a lubricant, a lubricant, a flavoring agent, an antioxidant, a buffer, a bacteriostatic agent, a diluent, a dispersant, a surfactant, Wherein the composition further comprises at least one adjuvant selected from the group consisting of: The method of claim 3,
The pharmaceutical composition for preventing or treating cancer may be selected from the group consisting of granules, powders, coated tablets, tablets, pills, capsules, suppositories, gels, syrups, juices, suspensions, emulsions, Wherein the pharmaceutical composition is for preventing or treating cancer. The method of claim 3,
Wherein said cancer is a solid cancer. 8. The method of claim 7,
The solid tumor may be selected from the group consisting of brain tumor, benign astrocytoma, malignant astrocytoma, pituitary adenoma, meningioma, brain lymphoma, oligodendroglioma, intracranial lesion, ependymoma, brain tumor, head and neck tumor, laryngeal cancer, Cancer, breast cancer, breast cancer, gastric cancer, liver cancer, gallbladder cancer, biliary cancer, pancreatic cancer, small bowel cancer, pancreatic cancer, breast cancer, pancreatic cancer, cancer, hypopharyngeal cancer, thyroid cancer, oral cancer, thoracic tumor, small cell lung cancer, non- Ovarian cancer, uterine sarcoma, vaginal cancer, female gynecomastia, female urethral cancer, ovarian cancer, ovarian cancer, ovarian cancer, uterine cancer, ovarian cancer, ovarian cancer, A pharmaceutical composition for the prevention or treatment of cancer which is skin cancer.

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