A 16-electron half-sandwich cationic complex with anticancer activity and its preparation method and application

Abstract

The present invention discloses a 16-electron half-sandwich cationic complex with anticancer activity, a preparation method and an application thereof. The structural formula of the complex is shown in formula (I), the ligand and the dimer react under reaction conditions, and after the inorganic salt AX is continuously added to the reaction system, a 16-electron half-sandwich cationic [N, NH] complex I is obtained, which is free of HCl compared to the common 18-electron complex; the role of AX is to serve as a counter ion to facilitate the precipitation of the cationic complex. This type of complex exhibits good anticancer activity, and its performance is close to or even better than that of commercial cisplatin, and is a very potential anticancer drug.

Description

Technical field:

[0001] The invention relates to a metal complex, in particular to a 16-electron half-sandwich cationic complex with anticancer activity and a preparation method and application thereof, belonging to the field of chemical pharmacy. Background technology:

[0002] Platinum drugs have achieved great success in the field of chemotherapy. The widespread use of cisplatin, carboplatin and oxaliplatin has promoted the rapid development of metal complexes in the field of anticancer. However, its serious side effects and poor selectivity have prompted people to turn their attention to other platinum metal anticancer drugs, such as Ru, Rh, Ir and Os complexes. Among them, semi-sandwich metal complexes have attracted widespread attention due to their easily adjustable structure, excellent anticancer activity and specific mechanism of action (MoAs).

[0003]

[0004] A large number of reported half-sandwich 18-electron complexes [(η 5 -Cp) / (η 6 -arene)M(XY)Cl] 0 / + (Cp = cyclopentadienyl functional group; M = Ru, Rh, Ir, Os; XY = bidentate ligand), adopts a hexacoordinated geometric structure (see structure, A), in which the chlorine atom, the bidentate ligand XY and η 5 -Cp / η 6 -arene occupies one coordination site, two coordination sites and three coordination sites respectively. However, there are few studies on the synthesis and anticancer activity of pentacoordinated 16-electron (without leaving group Cl) half-sandwich platinum group metal (including Rh, Ir, Ru and Os) complexes (see structure, B). The new special five-coordination mode of 16-electron half-sandwich complexes may increase the lipophilicity and cellular uptake of the complexes, thereby achieving better anticancer activity and being used to improve or supplement the performance of existing platinum drugs. Summary of the invention:

[0005] A series of [N,NH2] ligands with different substituents are introduced to obtain a series of novel 16-electron half-sandwich cationic [N,NH] complexes with good anti-tumor effects. Such novel 16-electron half-sandwich cationic [N,NH] complexes show excellent anti-tumor effects.

[0006]

[0007] The molecular structure of the half-sandwich cationic 16-electron [N, NH] complex is:

[0008]

[0009] In the formula, R1 and R2 are simultaneously or separately hydrogen, C1~C 15 Alkyl, halogen or halogenated C1~C 15 The alkyl group; wherein R3 is One of the following, X is Cl - PF6 - 、BF4 - ,BPh4 - 、SbF6 - , M is one of Ir, Rh, and Ru.

[0010] The chemical structural formulas of formula (I) of the present invention are as follows:

[0011] In formula (I), R1 is tert-butyl, R2 is hydrogen, and R3 is M is Ir, X is PF6 - The specific structural formula is shown in Formula 1; in Formula (I), R1 is methyl, R2 is methyl, and R3 is M is Ir, X is PF6 - The specific structural formula is shown in Formula 2; in Formula (I), R1 is tert-butyl, R2 is hydrogen, and R3 is M is Ru, X is PF6 - The specific structural formula is shown in Formula 3; in Formula (I), R1 is methyl, R2 is methyl, and R3 is M is Ru, X is PF6 - , the specific structural formula is shown in Formula 4;

[0012]

[0013] The preparation method of the 16-electron half-sandwich cationic [N, NH] complex of the present invention comprises the following steps: under a nitrogen atmosphere, a dimer represented by formula (III), a ligand represented by formula (II) and an inorganic salt AX are reacted in a methanol solvent at room temperature for 8 hours to obtain a 16-electron half-sandwich cationic complex represented by formula (I); the inorganic salt AX is one of NaX, KX, AgX and NH4X; the specific synthesis route is:

[0014]

[0015] The synthesis method of the four complexes 1-4 adopts the following steps:

[0016] When the complex is 1, it is prepared by the following method:

[0017] Under nitrogen atmosphere, 40.03 mg of iridium dimer (formula (III) R3 = M=Ir), 25.36mg (formula (Ⅱ) R2=hydrogen, R1=tert-butyl), 33mg ammonium hexafluorophosphate and 10mL anhydrous methanol, react at room temperature for 8h. After the reaction, the solution was dried by rotary evaporator, the solid was dissolved with dichloromethane, and the ammonium chloride and excess ammonium hexafluorophosphate were removed by sand core filtration; the filtrate was recrystallized by dichloromethane and n-hexane diffusion method, a precipitate appeared, the precipitate was filtered by sand core and washed with n-hexane, and dried in vacuo to obtain a red solid 1.

[0018] When the complex is 2, it is prepared by the following method:

[0019] Under nitrogen atmosphere, 40.03 mg of iridium dimer (formula (III) R3 = M=Ir), 23.95mg (formula (Ⅱ) R1=R2=methyl), 33mg ammonium hexafluorophosphate and 10mL anhydrous methanol, react at room temperature for 8h. After the reaction, the solution was dried by rotary evaporator, the solid was dissolved with dichloromethane, and the ammonium chloride and excess ammonium hexafluorophosphate were removed by sand core filtration; the filtrate was recrystallized by dichloromethane and n-hexane diffusion method, a precipitate appeared, the precipitate was filtered by sand core and washed with n-hexane, and dried in vacuo to obtain a red solid 2.

[0020] When the complex is 3, it is prepared by the following method:

[0021] Under nitrogen atmosphere, 29.26 mg of ruthenium dimer (formula (III) R3 = M=Ru), 24.12mg (formula (Ⅱ) R2=hydrogen, R1=tert-butyl), 33mg ammonium hexafluorophosphate and 10mL anhydrous methanol, react at room temperature for 8h. After the reaction, the solution was dried by rotary evaporator, the solid was dissolved with dichloromethane, and the ammonium chloride and excess ammonium hexafluorophosphate were removed by sand core filtration; the filtrate was recrystallized by dichloromethane and n-hexane diffusion method, a precipitate appeared, the precipitate was filtered by sand core and washed with n-hexane, and dried in vacuo to obtain a reddish brown solid 3.

[0022] When the complex is 4, it is prepared by the following method:

[0023] Under nitrogen atmosphere, 29.26 mg of ruthenium dimer (formula (III) R3 = M=Ru), 22.78mg (formula (Ⅱ) R1=R2=methyl), 33mg ammonium hexafluorophosphate and 10mL anhydrous methanol, react at room temperature for 8h. After the reaction, the solution was dried by rotary evaporator, the solid was dissolved with dichloromethane, and the ammonium chloride and excess ammonium hexafluorophosphate were removed by sand core filtration; the filtrate was recrystallized by dichloromethane and n-hexane diffusion method, a precipitate appeared, the precipitate was filtered by sand core and washed with n-hexane, and dried in vacuo to obtain a reddish brown solid 4.

[0024] Beneficial effects:

[0025] (1) The present invention provides a method for preparing a 16-electron half-sandwich cationic [N, NH] complex. A ligand, a dimer and an inorganic salt AX react under reaction conditions to obtain a novel 16-electron half-sandwich cationic complex I with good anti-tumor effect.

[0026] (2) These 16-electron half-sandwich cationic [N,NH] complexes 1-4 exhibit excellent anti-tumor effects, some of which are even better than commercial cisplatin, and are very promising anti-cancer drugs. Description of the drawings:

[0027] Figure 1 is the H NMR spectrum of complex 1 of the present invention.

[0028] Figure 2 is the mass spectrum of complex 1 of the present invention.

[0029] Figure 3 It is a single crystal of complex 1 of the present invention.

[0030] Figure 4 It is the H NMR spectrum of complex 2 of the present invention.

[0031] Figure 5 is the mass spectrum of complex 2 of the present invention.

[0032] Figure 6 It is a single crystal of complex 2 of the present invention.

[0033] Figure 7 It is the H NMR spectrum of complex 3 of the present invention.

[0034] Figure 8 is the mass spectrum of complex 3 of the present invention.

[0035] Fig. 9 is the H NMR spectrum of complex 4 of the present invention.

[0036] Fig.10 is the mass spectrum of complex 4 of the present invention. Specific implementation method:

[0037] The present invention is further illustrated by the following examples of some representative compounds, but these illustrations are not intended to limit the present invention.

[0038] The starting compounds used in the synthesis of the compounds are commercial products or can be prepared from known synthetic methods, and the preparation methods of all organic compounds can be obtained from the literature, and these methods are basic and obvious to synthetic chemists. Therefore, the description of the following synthetic methods can be considered to be detailed and specific.

[0039] Example 1

[0040] When the complex is 1, it is prepared by the following method:

[0041] Under nitrogen atmosphere, 40.03 mg of iridium dimer (formula (III) R3 = M=Ir), 25.36mg (formula (Ⅱ) R2=hydrogen, R1=tert-butyl), 33mg ammonium hexafluorophosphate and 10mL anhydrous methanol, react at room temperature for 8h. After the reaction, the solution was dried by rotary evaporator, the solid was dissolved with dichloromethane, and the ammonium chloride and excess ammonium hexafluorophosphate were removed by sand core filtration; the filtrate was recrystallized by dichloromethane and n-hexane diffusion method, a precipitate appeared, the precipitate was filtered by sand core and washed with n-hexane, and dried in vacuo to obtain a red solid 1.

[0042] NMR characterization 1 H NMR(500MHz,DMSO)δ9.32(s,1H,CH=N),7.93(d,1H),7.87(d,1H),7.77(t,J=7.6 Hz,1H),7.62(d,2H),7.31(m,3H),1.60(s,15H,Cp*-CH3),1.36(s,9H,C(CH3)3).

[0043] Mass Spectrum: C 27 H 34 The theoretical value of IrN2PF6 is 579.23512, the actual value is 579.23510, [M-PF6] + .

[0044] Elemental analysis. Theoretical value: C 27 H 34 IrN2PF6: C, 44.81; H, 4.74; N, 3.87. Actual measurement: C, 45.07; H, 4.61; N, 3.71.

[0045] Example 2

[0046] When the complex is 2, it is prepared by the following method:

[0047] Under nitrogen atmosphere, 40.03 mg of iridium dimer (formula (III) R3 = M=Ir), 23.95mg (formula (Ⅱ) R1=R2=methyl), 33mg ammonium hexafluorophosphate and 10mL anhydrous methanol, react at room temperature for 8h. After the reaction, the solution was dried by rotary evaporator, the solid was dissolved with dichloromethane, and the ammonium chloride and excess ammonium hexafluorophosphate were removed by sand core filtration; the filtrate was recrystallized by dichloromethane and n-hexane diffusion method, a precipitate appeared, the precipitate was filtered by sand core and washed with n-hexane, and dried in vacuo to obtain a red solid 2.

[0048] NMR characterization 1 H NMR (500MHz, CDCl3) δ8.68(s,1H,CH=N),8.04(m,1H),7.78(m,1H),7.56(m,1H),7.34(d,J=3. 8Hz,1H),7.13(s,2H),2.46(s,3H,Aryl-CH3),2.00(s,6H,Aryl-CH3),1.62(s,15H,Cp*-CH3).

[0049] Mass Spectrum: C 26 H 32 The theoretical value of IrN2PF6 is 565.21947, the actual value is 565.21838, [M-PF6] + .

[0050] Elemental analysis. Theoretical value: C 26 H 32 IrN2PF6: C, 44.00; H, 4.54; N, 3.95. Actual measurement: C, 44.34; H, 4.32; N, 3.67.

[0051] Example 3

[0052] When the complex is 3, it is prepared by the following method:

[0053] Under nitrogen atmosphere, 29.26 mg of ruthenium dimer (formula (III) R3 = M=Ru), 24.12mg (formula (Ⅱ) R2=hydrogen, R1=tert-butyl), 33mg ammonium hexafluorophosphate and 10mL anhydrous methanol, react at room temperature for 8h. After the reaction, the solution was dried by rotary evaporator, the solid was dissolved with dichloromethane, and the ammonium chloride and excess ammonium hexafluorophosphate were removed by sand core filtration; the filtrate was recrystallized by dichloromethane and n-hexane diffusion method, a precipitate appeared, the precipitate was filtered by sand core and washed with n-hexane, and dried in vacuo to obtain a reddish brown solid 3.

[0054] NMR characterization 1H NMR (500MHz, CDCl3) δ8.42(s,1H,CH=N),7.98(d,J=7.6Hz,1H),7.68(d,J=7.2Hz,3H),7.57(s,1H),7.49(d,3H),5.59(d,1H,arene-H),5.46(s,1H,aren e-H),5.38(d,1H,arene-H),5.23(s,1H,arene-H),2.77(m,1H,CH(CH3)2),1.95(s,3H,arene-CH3),1.39(s,9H,C(CH3)3),0.97(s,6H,CH(CH3)2). Mass spectrum: C 27 H 33 Theoretical value of RuN2PF6 is 487.16872, and the actual value is 487.16869, [M-PF6] +

[0055] Elemental analysis. Theoretical value: C 27 H 33 RuN2PF6: C, 51.34; H, 5.27; N, 4.44. Actual measured: C, 51.67; H, 5.09; N, 4.21.

[0056] Example 4

[0057] Under nitrogen atmosphere, 29.26 mg of ruthenium dimer (formula (III) R3 = M=Ru), 22.78mg (formula (Ⅱ) R1=R2=methyl), 33mg ammonium hexafluorophosphate and 10mL anhydrous methanol, react at room temperature for 8h. After the reaction, the solution was dried by rotary evaporator, the solid was dissolved with dichloromethane, and the ammonium chloride and excess ammonium hexafluorophosphate were removed by sand core filtration; the filtrate was recrystallized by dichloromethane and n-hexane diffusion method, a precipitate appeared, the precipitate was filtered by sand core and washed with n-hexane, and dried in vacuo to obtain a reddish brown solid 4.

[0058] NMR characterization 1H NMR (500MHz, CDCl3) δ8.52(s,1H,CH=N),8.11(d,J=8.6Hz,1H),7.65(t,J=7.2Hz,1H),7.51(m,2H),7.17(s,2H),5.64(d,2H,arene-H),5.51(d,J=6.3H z,2H,arene-H),2.49(s,3H,arene-CH3),2.38(m,1H,CH(CH3)2),2.04(s,6H,Aryl-CH3),2.02(s,3H,Aryl-CH3),1.15(d,J=6.9Hz,6H,CH(CH3)2). Mass spectrum: C 26 H 31 Theoretical value of RuN2PF6 is 473.15307, and the actual value is 473.15195, [M-PF6] +

[0059] Elemental analysis. Theoretical value: C 26 H 31 RuN2PF6: C, 50.57; H, 5.06; N, 4.54. Actual measurement: C, 50.78; H, 4.96; N, 4.32.

[0060] Example 5

[0061] Proliferation inhibition activity experiment of tumor cell lines of complexes 1-4 with anticancer activity:

[0062] (1) Preparation of test compounds: Dissolve the solid complex in DMSO to prepare a stock solution of a certain concentration, further dilute the stock solution with cell culture medium until the working concentration is reached, and culture for 24 h;

[0063] (2) Cell growth inhibition assay (MTT method):

[0064] 1) Take 5000 human cervical cancer cells (HeLa), human non-small cell lung cancer cells (A549) and human liver cancer cells (HepG2), prepare cell suspensions, and inoculate them in 96-well culture plates;

[0065] 2) Pre-culture cells with drug-free medium, incubate at 5% CO2, 310K for 24 hours, add the prepared test compound, and culture for 24 hours;

[0066] 3) Add 15 μL of 5 mg / mL MTT solution to each well and continue culturing for 4 hours to form a purple crystalline substance, formazan;

[0067] 4) Terminate the culture, carefully remove the culture medium in the wells, add 100 μL of DMSO to each well to fully dissolve the formazan precipitate, mix on an oscillator, and measure the optical density of each well with a microplate reader at a wavelength of 570 nm;

[0068] 5) Each experiment was repeated three times, IC 50 = Mean ± SEM. The inhibition rates of complexes 1-4 and commercial cisplatin on the growth of cancer cells HeLa, A549 and HepG2 are shown in Table 1.

[0069] Table 1

[0070]

[0071] It can be seen from Example 5 that complexes 1-4 all show very good anticancer activity, which is close to or better than the activity of commercialized cisplatin. The anticancer activity of complex 1 is different from that of complex 3, proving that metal has an effect on the anticancer activity. The anticancer activity of complex 2 is also better than that of complex 1, proving that the anticancer activity changes with the change of the substituent. The above conclusions provide a theoretical basis for the preparation of new anti-tumor drugs.

[0072] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. 16 electron half sandwich cationic [N,NH] complex, characterized in that: The structural formula is shown in formula (I): ； In the formula (I), R1 is tert-butyl, R2 is hydrogen, and R3 is ,M is Ir,X is PF6 - The specific structural formula is shown in Formula 1; in Formula (I), R1 is methyl, R2 is methyl, and R3 is ,M is Ir,X is PF6 - The specific structural formula is shown in Formula 2; in Formula (I), R1 is tert-butyl, R2 is hydrogen, and R3 is ,M is Ru, X is PF6 - The specific structural formula is shown in Formula 3; in Formula (I), R1 is methyl, R2 is methyl, and R3 is ,M is Ru, X is PF6 - , the specific structural formula is shown in Formula 4; 。 2. A method for preparing the 16-electron half-sandwich cationic [N,NH] complex according to claim 1, characterized in that: The following steps are involved: Under nitrogen atmosphere, the dimer represented by formula (III), the ligand represented by formula (II) and the inorganic salt AX are reacted in the solvent of methanol at room temperature for 8 h to obtain the half sandwich cationic 16-electron complex represented by formula (I); the inorganic salt AX is one of NaX, KX, AgX and NH4X; the specific synthesis route is: 。 3. The preparation method according to claim 2, characterized in that: When the complex is 1, it is prepared by the following method: Under nitrogen atmosphere, 40.03 mg of iridium dimer of formula (III), 25.36 mg of formula (II), 33 mg of ammonium hexafluorophosphate and 10 mL of anhydrous methanol were added to a 100 mL Schlenk bottle, wherein R3 of formula (III) = , M = Ir, R2 = hydrogen, R1 = tert-butyl in formula (II), react at room temperature for 8 h, after the reaction, the solution is dried by rotary evaporator, the solid is dissolved by dichloromethane, and the ammonium chloride and excess ammonium hexafluorophosphate are removed by sand core filtration; the filtrate is recrystallized by dichloromethane and n-hexane diffusion method, a precipitate appears, the precipitate is filtered by sand core and washed with n-hexane, and vacuum dried to obtain a red solid complex 1; When the complex is 2, it is prepared by the following method: Under nitrogen atmosphere, 40.03 mg of iridium dimer of formula (III), 23.95 mg of formula (II), 33 mg of ammonium hexafluorophosphate and 10 mL of anhydrous methanol were added to a 100 mL Schlenk bottle, wherein R3 of formula (III) = , M=Ir, formula (II) R1=R2=methyl, react at room temperature for 8 h, after the reaction, the solution was dried by rotary evaporator, the solid was dissolved with dichloromethane, and the ammonium chloride and excess ammonium hexafluorophosphate were removed by sand core filtration; the filtrate was recrystallized by dichloromethane and n-hexane diffusion method, and a precipitate appeared. The precipitate was filtered by sand core and washed with n-hexane, and vacuum dried to obtain a red solid complex 2; When the complex is 3, it is prepared by the following method: Under nitrogen atmosphere, 29.26 mg of ruthenium dimer of formula (III), 24.12 mg of formula (II), 33 mg of ammonium hexafluorophosphate and 10 mL of anhydrous methanol were added to a 100 mL Schlenk bottle, wherein R3 of formula (III) = , M=Ru, formula (II) R2=hydrogen, R1=tert-butyl, react at room temperature for 8 h. After the reaction, the solution was dried by rotary evaporator, the solid was dissolved with dichloromethane, and the ammonium chloride and excess ammonium hexafluorophosphate were removed by sand core filtration. The filtrate was recrystallized by dichloromethane and n-hexane diffusion method, and a precipitate appeared. The precipitate was filtered by sand core and washed with n-hexane, and vacuum dried to obtain a reddish brown solid complex 3. When the complex is 4, it is prepared by the following method: Under nitrogen atmosphere, 29.26 mg of ruthenium dimer of formula (III), 22.78 mg of formula (II), 33 mg of ammonium hexafluorophosphate and 10 mL of anhydrous methanol were added to a 100 mL Schlenk bottle, wherein R3 of formula (III) = , M=Ru, formula (II) R1=R2=methyl, react at room temperature for 8 h. After the reaction, the solution was dried by rotary evaporator, the solid was dissolved with dichloromethane, and the ammonium chloride and excess ammonium hexafluorophosphate were removed by sand core filtration. The filtrate was recrystallized by dichloromethane and n-hexane diffusion method, and a precipitate appeared. The precipitate was filtered through a sand core and washed with n-hexane, and dried in vacuo to obtain a reddish-brown solid complex 4.

4. Use of the 16-electron half-sandwich cationic [N, NH] complex according to claim 1 in the preparation of anticancer drugs, wherein the cancer is liver cancer, lung cancer or cervical cancer.

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