Platinum (IV) complex
10988496 · 2021-04-27
Assignee
Inventors
Cpc classification
C07F15/00
CHEMISTRY; METALLURGY
International classification
Abstract
According to the present invention, there is a demand for novel platinum(IV) complex which has sufficient water-solubility, stability, antitumor activity, and the like that are required as a medicine, and may be used clinically. Thus, provided is a platinum(IV) complex represented by the following General Formula (I) [wherein X.sub.1 and X.sub.2 each represent a halogen atom or are bonded together to form a dicarboxylate selected from the group consisting of oxalate, malonate, succinate, and o-phthalate; and Y represents a halogen atom. ##STR00001##
Claims
1. An isolated platinum(IV) complex of the following General Formula (I): ##STR00006## wherein X.sub.1 and X.sub.2 form oxalate and Y represents a chlorine atom or a bromine atom.
2. The isolated platinum(IV) complex according to claim 1, wherein the 1,2-cyclohexanediamine ligand is a (1R,2R)-cyclohexanediamine ligand.
3. A medicine comprising the isolated platinum(IV) complex according to claim 1 as an active ingredient.
4. A medicine comprising the isolated platinum(IV) complex according to claim 2 as an active ingredient.
Description
BRIEF DESCRIPTION OF DRAWINGS
(1)
DETAILED DESCRIPTION OF THE INVENTION
(2) Hereinafter, the details of the present invention will be described.
(3) The halogen atom according to the present invention is a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom. It is preferable that X.sub.1 and X.sub.2 both represent the same halogen atom, and above all, it is particularly preferable that X.sub.1 and X.sub.2 both represent a chlorine atom or a bromine atom. Y is preferably a chlorine atom or a bromine atom.
(4) The dicarboxylate, which is a leaving group, according to the present invention is not particularly limited, and examples include a (C1-C6) alkyl group having two carboxyl groups, and a (C6-C10) aryl group having two carboxyl groups. Among them, oxalate, malonate, succinate, and o-phthalate shown below are preferred.
(5) ##STR00003##
(6) The platinum complex of the present invention is particularly preferably a compound represented by the following General Formula (II) or General Formula (IV).
(7) ##STR00004##
wherein Y represents a halogen atom.
(8) Regarding the steric structure of the 1,2-cyclohexanediamine ligand of the platinum(IV) complex of the present invention, (1R, 2R)-trans disposition is preferred from the viewpoint of physiological activity or the like.
(9) That is, the platinum(IV) complex of the present invention is particularly preferably a compound represented by the following General Formula (III) or General Formula (V).
(10) ##STR00005##
wherein Y represents a halogen atom.
(11) The platinum(IV) complex of the present invention may be produced by applying methods described is the literature such as Non Patent Literature 2. That is, a method of treating a platinum(II) complex with an oxidizing agent such as hydrogen peroxide, or subjecting a platinum(II) complex to an oxidative halogenation, and thereby obtaining an intended platinum(IV) complex; or a method of subjecting a platinum(IV) complex to a substitution reaction, and thereby obtaining an intended platinum(IV) complex, may be used. Examples of these production methods are described in the following Examples.
(12) A medicine including the platinum(IV) complex of the present invention as an active ingredient is also included in the present invention. The pharmaceutical application in which the platinum(IV) complex of the present invention exhibits efficacy is not particularly limited; however, a use application as an anticancer agent is preferred. Regarding the use as an anticancer agent, the platinum(IV) complex may be used alone, or may be mixed with pharmaceutically acceptable additives such as a carrier, an excipient, a disintegrant, a binder, a lubricating agent, a fluidizing agent, a coating agent, a suspending agent, an emulsifier, a stabilizer, a preservative, a flavoring agent, a fragrance, a diluents, and a dissolution aid. The anticancer agent may be administered orally or parenterally (systemic administration, topical administration, or the like) in the form of preparations such as a powder preparation, a granular preparation, a tablet, a caplet, a capsule, an injectable preparation, a suppository, and an ointment. The platinum(IV) complex of the present invention in the preparation may vary widely depending on the preparation; however, the proportion is usually 0.1% to 100% by weight. The dose may vary depending on the route of administration, the age of the patient, the actual symptoms to be prevented or treated, and the like; however, for example, in the case of administering the preparation to an adult, the platinum(IV) complex may be administered, as an active ingredient, at a dose of 0.01 mg to 2,000 mg, and preferably 0.1 mg, to 1,000 mg, per day, and may be administered once a day or in several divided portions a day.
EXAMPLES
(13) Hereinafter, the present invention will be described in more detail by way of Examples. However, the present invention is not intended to be limited to these Examples.
(14) In the Examples of the present invention, the following abbreviations will be used.
(15) ox: oxalate
(16) cbdc: 1,1-cyclobutanedicarboxylate
(17) 1-OHP: oxaliplatin
(18) Measurement of the purity of compounds in the present Example was carried out by using high performance liquid chromatography and using L-column2 ODS (4.6 mm I.D.×250 mm; purchased from Chemicals Evaluation and Research Institute, Japan) as a column; a buffer solution prepared by dissolving 2.72 g of potassium dihydrogen phosphate, 1.89 g of sodium 1-pentanesulfonate, and 0.5 ml of triethylamine in 2,000 ml of distilled water and adjusting the solution to pH 4.3 with phosphoric acid, as a mobile phase (A); and methanol as a mobile phase (B), under the following analysis conditions 1 or 2.
(19) Analysis Conditions 1 (Isocratic Analysis):
(20) Mobile phase (B) concentration: 15% (0 min) to 15% (20 min)
(21) Mobile phase flow rate: 1 ml/min, detection: 210 nm
(22) Analysis Conditions 2 (Gradient Analysis):
(23) Mobile phase (3) concentration: 15% (0 min) to 90% (10 min)
(24) Mobile phase flow rate: 1 ml/min, detection 210 nm.
Example 1
Synthesis of trans,cis,cis-[PtCl(OH) (R,R-dach) (ox)]: Y═Cl in General Formula (III)
(25) N-chlormide (66.8 mg) was dissolved in 14 ml of distilled water, and a liquid obtained by suspending 1-OHP (200 mg) in 6 ml of distilled water was added thereto. The mixture thus obtained was stirred for 4 hours at room temperature in the dark. After completion of the reaction, insoluble materials in the reaction liquids were separated by filtration, the filtrate was concentrated under reduced pressure, and thereby a solid was obtained. The solid thus obtained was recrystallized from ethanol/water, and thus the title compound (114 mg) was obtained. .sup.1H-NMR (D.sub.2O): δ 2.89-2.72 (2H, m), 2.15 (2H, d, J=12.2 Hz), 1.53-1.41 (4H, m), 0.97-0.90 (2H, m), MS (ESI; Electrospray Ionization): 450 (M+1), 451 (M+2), purity (HPLC, analysis conditions 2): 99.4%.
Example 2
Synthesis of trans,cis,cis-[PtBr(OH) (R,R-dach) (ox)]: Y═Br in General Formula (III)
(26) N-bromosuccinmide (89.6 mg) was dissolved in 14 ml of distilled water, a liquid obtained by suspending 1-OHP (200 mg) in 6 ml of distilled water was added thereto. The mixture thus obtained was stirred for 3 hours at room temperature in the dark. After completion of the reaction, insoluble materials in the reaction liquid were separated by filtration, the filtrate was concentrated under reduced pressure, and thereby a solid was obtained. The solid thus obtained was suspended in water and collected by filtration again, and thus the title compound (216 mg) was obtained. .sup.1H-NMR (DMSO-d.sub.6): δ 7.91-7.65 (2H, m) 7.14-7.03 (2H, m), 2.65-2.55 (2H, m), 2.07-1.94 (2H, m), 1.50-1.46 (4H, m), 1.15-1.02 (2H, m), MS (ESI): 495 (M+1), purity (HPLC, analysis conditions 2): 98.9%.
Example 3
Synthesis of trans,cis,cis-[PtCl(OH) (R,R-dach) (Cl).SUB.2.]: X.SUB.1., X.SUB.2., Y═Cl in General Formula (I)
(27) N-chlorosuccinimide (105.4 mg) was dissolved in 7 ml of distilled water, the solution was added to a liquid obtained by suspending Pt (R,R-dach) Cl.sub.2 (300 mg) in 60 ml of tetrahydrofuran. The mixture thus obtained was stirred for 4 hours at room temperature in the dark. After completion of the reaction, insoluble materials in the reaction liquid were separated by filtration, the filtrate was concentrated under reduced pressure, and thereby a solid was obtained. The solid thus obtained was suspended in ethanol and collected by filtration again, and thus the title compound (322 mg) was obtained. .sup.1H-NMR (DMSO-d.sub.6): δ 7.53-7.29 (2H, m), 6.89-6.78 (2H, m), 2.75-2.60 (2H, m), 2.10-2.00 (2H, m), 1.47 (2H, d, J=8.0 Hz), 1.10-0.93 (2H, m), MS (PSI): 433 (M+1), purity (HPLC, analysis conditions 2): 98.1%.
Comparative Example 1
Synthesis of trans,cis,cis-[Pt(OH)(OAc)(R,R-dach) (ox)]
(28) 0.135 ml of a 30% aqueous solution of hydrogen peroxide was added to a liquid obtained by suspending 1-OHP (200 mg) in 9 ml of acetic acid. The mixture thus obtained was stirred for 19 hours at room temperature in the dark. After completion of the reaction, the mixture was concentrated under reduced pressure several times while water added thereto, and thus a solid was obtained. The solid thus obtained was recrystallized from ethanol/methanol, and thus the title compound (55 mg) was obtained. .sup.1H-NMR (D.sub.2O): δ 2.78-2.73 (2H, m), 2.17 (2H, d, J=9.2 Hz), 1.94 (3H, s), 1.54-1.44 (4H, m), 1.20-1.05 (2H, m) purity (HPLC, analysis conditions 1): 94.0).
Comparative Example 2
Synthesis of trans,cis,cis-[PtCl (OCH.SUB.2.CH.SUB.2.OH) (R,R-dach) (ox)]
(29) N-chlorosuccinimide (66.8 mg) was added to a liquid obtained by suspending 1-OHP (200 mg) in 2 ml of ethylene glycol. The mixture thus obtained was stirred for 3 hours at room temperature in the dark. After completion of the reaction, 10 ml of acetone and 30 ml of diethyl ether were added to the reaction liquid, and a solid precipitated therefrom was collected by filtration. The solid thus obtained was recrystallized from ethanol/water, and thus the title compound (154 mg) was obtained. .sup.1H-NMR (D.sub.2O): 53.58-3.45 (2H, m), 3.22-3.08 (2H, m), 2.85-2.83 (2H, m), 2.14 (2H, d, J=11.2 Hz), 1.53-1.44 (4H, m), 1.15-1.07 (2H, m), purity (HPLC, analysis conditions 1): 98.0%.
Comparative Example 3
Synthesis of trans,cis,cis-[Pt(OH).SUB.2 .(R,R-dach) (ox)]
(30) 2.58 ml of a 30% aqueous solution of hydrogen peroxide was added to a liquid obtained by suspending 1-OHP (900 mg) in 12 ml of distilled water. The mixture thus obtained was stirred for 20.5 hours at room temperature in the dark. After completion of the reaction, the mixture was concentrated under reduced pressure several times while water was added thereto, and a solid was obtained. The solid thus obtained was recrystallized from distilled water, and thereby the title compound (422 mg) was obtained. .sup.1H-NMR (D.sub.2O): δ 2.74-2.72 (2H, m), 2.17 (2H, d, J=12.8 Hz), 1.54-1.45 (4H, m), 1.18-1.12 (2H, m), purity (HPLC, analysis conditions 1): >98.0%.
Comparative Example 4
Synthesis of trans,cis,cis-[Pt(OCOCH.SUB.2.CH.SUB.2.C.SUB.6.H.SUB.5.).SUB.2 .(R,R-dach)(ox)]
(31) 3-Phenylpropionic acid (77 mg) and N,N-dimethylaminopyridine (5.7 mg) were dissolved in 2 ml of N,N-dimethylformamide, 0.086 ml of diisopropylcarbodiimide was added thereto, and then the mixture was stirred for 0.5 hours at room temperature. To the reaction liquid, a liquid obtained by suspending trans,cis,cis-[Pt(OH).sub.2 (R,R-dash)(ox)] (200 mg) obtained in Comparative Example 3 in 2 ml of N,N-dimethylformamide was added. The mixture thus obtained was stirred for 23 hours at room temperature in the dark. The reaction liquid was filtered to exclude any unreacted platinum complex, and a solid was precipitated by adding water to the filtrate thus obtained. The solid was collected by filtration and was washed with cold ethanol, and thus the title compound (38 mg) was obtained. .sup.1H-NMR (DMSO-d.sub.6): δ 8.30 (4H, brs), 7.27-7.14 (10H, m), 2.80-2.76 (4H, m), 2.60-2.56 (4H, m), 2.40-2.30 (2H, m), 2.05 (2H, d, J=12.4 Hz), 1.47 (2H, d, J=8.0 Hz), 1.40-1.22 (2H, m), 1.15-1.14 (2H, m), purity (HPLC, analysis conditions 2): 98.0%.
Comparative Example 5
Synthesis of trans,cis,cis-[PtCl(OH) (R,R-dach) (cbdc)]
(32) The title compound was synthesized according to the method described in Non Patent Literature 3. .sup.1H-NMR (DMSO-d.sub.6): δ 7.71-7.43 (2H, m), 7.00-6.90 (2H, m), 2.60-2.29 (6H, m), 2.03-1.93 (2H, m), 1.84-1.49 (2H, m), 1.50-1.30 (4H, m), 1.05-0.95 (2H, m), MS (ESI): 504 (M+1), 486 (M+OH), purity (HPLC, analysis conditions 2): 95.6%.
Comparative Example 6
Synthesis of trans,cis,cis-[Pt(OH).SUB.2.(R,R-dach) (cbdc)]
(33) Cis,cis-[Pt(R,R-dach) (cbdc)] (100 mg) synthesized according to the method described in Non Patent Literature 3 was dissolved in 14 ml of a 50% acetone solution, 14 ml of a 30% aqueous solution of hydrogen peroxide was added thereto. The mixture thus obtained was stirred for 4 hours at room temperature in the dark. After completion of the reaction, the mixture was concentrated under reduced pressure several times while water was added thereto, and a solid was obtained. The solid thus obtained was suspended and purified in acetone, and thus the title compound (41 mg) was obtained. .sup.1H-NMR (D.sub.2O): 2.97 (2H, d, J=10.0 Hz), 2.77-2.72 (4H, m), 2.36-2.32 (2H, m), 2.14-2.10 (2H, m), 1.74-1.64 (4H, m) 1.37-1.34 (2H, m), MS (ESI): 486 (M+1), 486 (M−OH), purity (HPLC, analysis conditions 2): 96.8%.
Test Example 1
In Vitro Antitumor Assay for Example Compounds and Comparative Example Compounds
(34) Gastric cancer and pancreatic cancer cell lines were respectively inoculated on a 96-well plate. Gastric cancer cells KATO III were inoculated at a rate of 1×10.sup.4 cells/well gastric cancer cells MKN-1 were inoculated at a rate of 5×10.sup.5 cells/well, gastric cancer cells MKN-45 were inoculated at a rate of 1×10.sup.4 cells/well, gastric cancer cells MKN-74 were inoculated at a rate of 1×10.sup.4 cells/well, pancreatic cancer cells AsPC-1 were inoculated at a rate of 5×10.sup.7 cells/well, pancreatic cancer cells BxPC-3 were inoculated at a rate of 5×10.sup.5 cells/well, pancreatic cancer cells DAN-G were inoculated at a rate of 5×10.sup.5 cells/well, and pancreatic cancer cells SUIT-2 were inoculated at a rate of 5×10.sup.5 cells/well. After culturing the cells for 24 hours, each of the Example compounds or each of the Comparative Example compounds was added to the cells to obtain a final concentration of from 0.0244 μmol/L to 100 μmol/L at a common ratio of 4. Three technical replicates were used. Wells to which no drug was added were prepared as control, and wells to which cells and drugs were not added were prepared as blanks. After the cells were cultured for 72 hours, the culture fluid was removed, the cells were fixed with methanol, and then the cells were stained using a Methylene Blue stain solution. After excess Methylene blue stain solution was washed off, 200 μL of 0.1% hydrochloric acid was added to each well, and the dye was extracted. The light absorbance at 660 nm was measured using a microplate reader, and the cell proliferation inhibitory activity (GI %) was calculated from the light absorbance thus obtained by the following formula.
GI.sub.XY %=(1−(A.sub.XY−B)/(C−B))×100
(35) Here, GI.sub.XY % represents the cell prolj feration-inhibitory activity when the concentration of compound. X is Y μM; A.sub.XY represents the average light absorbance of the well to which compound X has been added at Y μM; B represents the light absorbance of a blank well; and C represents the tight absorbance of a control well.
(36) The GI.sub.XY % was determined for various compound concentrations, and a proliferation-inhibition curve was plotted from the concentration and the cell proliferation-inhibitory activity. Thus, the concentration at which the cell proliferation-inhibitory activity was 50% was designated as the IC.sub.50 value of compound X. The results are presented in Tables 1, 2, and 3.
(37) TABLE-US-00001 TABLE 1 Axial ligand Cell line IC.sub.50 (μM) Y X.sub.1, X.sub.2 KATO III MKN-1 MKN-45 MKN-74 Compound of Example 1 OH, Cl ox 13.8 1.9 0.3 16 Compound of Comparative Example 1 OH, OAc ox 55.7 45.3 6.2 >100 Compound of Comparative Example 2 OC.sub.3H.sub.4OH, Cl ox 42.6 10.8 1.9 58.1 Compound of Comparative Example 3 OH, OH ox n.t >100 n.t >100 Compound of Comparative Example 4 OCOR, OCOR ox n.t 9.2 n.t 27.3 1-OHP — ox 9.2 0.5 0.1 8.6 Pt(dach)Cl.sub.2 — Cl, Cl 13.8 3.7 0.5 22.3 R represents CH.sub.2CH.sub.2C.sub.6H.sub.5. n.t. stands for “not tested”.
(38) TABLE-US-00002 TABLE 2 Axial ligand Cell line IC.sub.50 (μM) Y X.sub.1, X.sub.2 BxPC-3 SUIT-2 DAN-G AsPC-1 Compound of Example 1 OH, Cl ox 1.2 0.9 1 3.1 Compound of Comparative Example 1 OH, OAc ox 44.3 26.1 36.7 62.9 Compound of Comparative Example 2 OC.sub.3H.sub.4OH, Cl ox 5.7 6.1 5.6 9.7 Compound of Comparative Example 3 OH, OH ox n.t 39.2 >100 n.t. Compound of Comparative Example 4 OCOR, OCOR ox n.t 1 6.8 n.t. 1-OHP — ox 0.8 0.4 1.1 1.6 Pt(dach)Cl.sub.2 — Cl, Cl 3.4 2.1 8.5 14.7 R represents CH.sub.2CH.sub.2C.sub.6H.sub.5.
(39) Example 1 compound exhibited high antitumor effects against all cell lines, compared to the compounds of Comparative examples 1 to 4, in which the combinations of the axial ligands were different. From this, it became clear that regarding the combination of axial ligands in a platinum(IV) complex having the dach structure, the combination of a hydroxyl group and a halogen atom of the compound of Example 1 was excellent. Meanwhile, the compound of Example 1 exhibited an activity equivalent to that of 1-OHP that is used as an anticancer agent, and exhibited higher activity compared to Pt (dach) Cl.sub.2.
(40) TABLE-US-00003 TABLE 3 Ligand Cell line IC.sub.50 (μM) Axial X.sub.1, X.sub.2 MKN-1 MKN-74 SUIT-2 DAN-G Compound of Example 1 OH, Cl ox 1.9 16 0.9 1 Compound of Example 2 OH, Br ox 2.7 n.t. 1.5 n.t. Compound of Example 3 OH, Cl Cl, Cl 2.6 17.1 0.9 8.6 Compound of Comparative Example 3 OH, OH ox >100 >100 39.2 >100 Compound of Comparative Example 5 OH, Cl cbdc 4.5 29 3.3 3.4 Compound of Comparative Example 6 OH, OH cbdc >100 >100 79.4 >100 1-OHP — ox 0.5 8.6 8.4 1.1
(41) Although the compounds of Comparative Examples 5 and 6 had inferior activity compared to the compounds of the present invention, it became clear from the results for the compound of Comparative Example 5 and the compound of Comparative Example 6 that even in a case in which X.sub.1 and X.sub.2 were converted to cbdc, regarding the combination of the axial ligands of the platinum(IV) complex having the dach structure, the combination of a hydroxyl group and a chlorine atom or a bromine atom is excellent. Furthermore, it became clear from the results for the compound of Example 3 that a combination in which Y represents a chlorine atom, and X.sub.1 and X.sub.2 both represent a chlorine atom also exhibits high antitumor activity.
Test Example 2
Test on Solubility in Water of Compound of Example 1 and Compound of Comparative Example 5
(42) The compound of Example 1 and the compound of Comparative Example 5 were weighed, distilled water was slowly added to each of the compounds, and thereby the concentration at which crystals were completely dissolved was measured. The results are presented in Table 4. The solubility of 1-OHP is the reference value calculated from the literature value.
(43) TABLE-US-00004 TABLE 4 Solubility (mg/ml) Compound of Example 1 7 Compound of Comparative Example 5 3 l-OHP 2-2.5
(44) As a result, it became clear that the solubility in water of the compound of Example 1, which is a platinum(IV) complex having a hydroxyl group and a halogen atom introduced thereinto as the axial ligands of the present invention, increased by about 3 times the solubility of 1-OHP, which is a corresponding platinum(II) complex. Furthermore, the solubility was higher by two times or more than that of the compound of Comparative Example 5, which is an existing platinum(IV) complex.
Test Example 3
Test on Solution Stability in Distilled Water of Compound of Example 1 and Compound of Comparative Example 5
(45) The compound of Example 1 and the compound of Comparative Example 5 were weighed in a vessel, and the compounds were dissolved to a concentration of 1 mg/ml using distilled water. Each of the aqueous solutions was filtered using a syringe filter having a pore size of 0.45 μm, and the filtrate was shaken in a water bath at 37° C. in the dark. Sampling was performed over time, and stability was tested by high performance liquid chromatography. The results are presented in
(46) As a result of the test, the residual ratio of the compound of Example 1 after 74 hours was 99.1%, while the residual ratio of the compound of Comparative Example 5, which is an existing platinum(IV) complex, was 63.7%. It is obvious that the compound of Example 1 of the present invention was stable for a long time period in an aqueous solution and was stable even compared to the compound of Comparative Example 5.
Test Example 4
Test on Solution Stability Physiological Saline of Compound of Example 1
(47) The compound of Example 1 was weighed in a vessel, and the compound was dissolved to a concentration of 1 mg/ml using physiological saline. The solution was allowed to stand at 5° C. in the dark or was shaken in a water bath at 37° C. without blocking light, and the residual amount was quantitatively determined by high performance liquid chromatography. The residual ratio is presented in Table 5.
(48) TABLE-US-00005 TABLE 5 Residual ratio Conditions 3 hours 24 hours Compound of 5° C., in the dark, allowed to stand 100% 98.77% Example 1 Compound of 37° C., without blocking light, 100% 94.43% Example 1 shaken
(49) Generally, a platinum complex having a leaving group other than a chlorine atom, for example, 1-OHP, undergoes exchange of chlorine ions in physiological saline, and therefore, the platinum complex is unstable in physiological saline. However, as shown by the results of the present test, the compound of Example 1 of the present invention, which is a platinum(IV) complex having a dicarboxylate as a leaving group, almost does not undergo decomposition after 24 hours at 5° C. in the dark even in physiological saline. Even though the compound of Example 1 was shaken at 37° C. without blocking light, which constituted more severe conditions, the residual ratio was 94.4%, and the compound was stable even in physiological saline.
(50) From the various test results described above, it has become clear that the platinum complex of the present invention has excellent antitumor activity and excellent solubility, and has excellent performance that even if the platinum complex is produced into a solution, the platinum complex is chemically stable.