Antitumor Complexes Research Articles

Platinum Antitumor Complexes: 50 Years Since Barnett Rosenberg's Discovery.

Author(s): Muggia, Franco M; Bonetti, Andrea; Hoeschele, James D; Rozencweig, Marcel; Howell, Stephen B

Nucleolar targeting by platinum: p53-independent apoptosis follows rRNA inhibition, cell-cycle arrest, and DNA compaction.

TriplatinNC is a highly positively charged, substitution-inert derivative of the phase II clinical anticancer drug, BBR3464. Such substitution-inert complexes form a distinct subset of polynuclear platinum complexes (PPCs) interacting with DNA and other biomolecules through noncovalent interactions. Rapid cellular entry is facilitated via interaction with cell surface glycosoaminoglycans and is a mechanism unique to PPCs. Nanoscale secondary ion mass spectrometry (nanoSIMS) showed rapid distribution within cytoplasmic and nucleolar compartments, but not the nucleus. In this article, the downstream effects of nucleolar localization are described. In human colon carcinoma cells, HCT116, the production rate of 47S rRNA precursor transcripts was dramatically reduced as an early event after drug treatment. Transcriptional inhibition of rRNA was followed by a robust G1 arrest, and activation of apoptotic proteins caspase-8, -9, and -3 and PARP-1 in a p53-independent manner. Using cell synchronization and flow cytometry, it was determined that cells treated while in G1 arrest immediately, but cells treated in S or G2 successfully complete mitosis. Twenty-four hours after treatment, the majority of cells finally arrest in G1, but nearly one-third contained highly compacted DNA; a distinct biological feature that cannot be associated with mitosis, senescence, or apoptosis. This unique effect mirrored the efficient condensation of tRNA and DNA in cell-free systems. The combination of DNA compaction and apoptosis by TriplatinNC treatment conferred striking activity in platinum-resistant and/or p53 mutant or null cell lines. Taken together, our results support that the biological activity of TriplatinNC reflects reduced metabolic deactivation (substitution-inert compound not reactive to sulfur nucleophiles), high cellular accumulation, and novel consequences of high-affinity noncovalent DNA binding, producing a new profile and a further shift in the structure–activity paradigms for antitumor complexes.

Condensations of single DNA molecules induced by heptaplatin and its chiral isomer

Heptaplatin is a third-generation platinum antitumor drug. It has a chiral isomer. We studied the interactions between the two isomers and DNA by using magnetic tweezers and atomic force microscopy (AFM) to investigate the effect of chiralities of the isomers on the interactions. We found that the extension curves and average condensation rates of DNA molecules incubated with heptaplatin were nearly the same as those incubated with its chiral isomer. In addition, the structures of DNA molecules incubated with heptaplatin were also similar to those incubated with its chiral isomer. These results indicate the difference in chirality of the two isomers does not induce different interactions of the isomers with DNA. Our study may facilitate the understanding of interactions of platinum complexes with DNA and the design of new antitumor platinum complexes.

Anticancer activity and DNA-binding properties of novel cationic Pt(II) complexes

In this study, three structurally related cationic Pt complexes, [Pt(ppy)(dppe)]CF3CO2: C1, [Pt(bhq)(dppe)]CF3CO2: C2, and [Pt(bhq)(dppf)]CF3CO2: C3, in which ppy=deprotonated 2-phenylpyridine, bhq=deprotonated benzo[h]quinoline, dppe=bis(diphenylphosphino)ethane and dppf=1,1′-bis(diphenylphosphino)ferrocene, were used for the assessment of their anticancer activities against Jurkat and MCF-7 cancer cell lines. The Pt complexes (C1–C3) demonstrated significant level of anticancer properties, as measured using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Moreover, the changes in nuclear morphology with Acridine Orange (AO) staining reveal that these complexes are capable to induce apoptosis, and only C1 stimulates activity of Caspase-3 in Jurkat cancer cells. To get a better insight into the nature of binding between these cationic Pt complexes and DNA, different spectroscopic techniques and gel electrophoresis were applied. On the basis of the results of UV/vis absorption spectroscopy, CD experiment and fluorescence quenching of ethidium bromide (EB)–DNA, the interaction between DNA and the Pt complexes is likely to occur through a mixed-binding mode. Overall, the present work suggests that a controlled modification could result in new potentially antitumor complexes which can survive the repair mechanism and induce facile apoptosis.

Bovine lactoferrin binds oleic acid to form an anti-tumor complex similar to HAMLET

α-Lactalbumin (α-LA) can bind oleic acid (OA) to form HAMLET-like complexes, which exhibited highly selective anti-tumor activity in vitro and in vivo. Considering the structural similarity to α-LA, we conjectured that lactoferrin (LF) could also bind OA to obtain a complex with anti-tumor activity. In this study, LF–OA was prepared and its activity and structural changes were compared with α-LA–OA. The anti-tumor activity was evaluated by methylene blue assay, while the apoptosis mechanism was analyzed using flow cytometry and Western blot. Structural changes of LF–OA were measured by fluorescence spectroscopy and circular dichroism. The interactions of OA with LF and α-LA were evaluated by isothermal titration calorimetry (ITC). LF–OA was obtained by heat-treatment at pH8.0 with LD50 of 4.88, 4.95 and 4.62μM for HepG2, HT29, and MCF-7 cells, respectively, all of which were 10 times higher than those of α-LA–OA. Similar to HAMLET, LF–OA induced apoptosis in tumor cells through both death receptor- and mitochondrial-mediated pathways. Exposure of tryptophan residues and the hydrophobic regions as well as the loss of tertiary structure were observed in LF–OA. Besides these similarities, LF showed different secondary structure changes when compared with α-LA, with a decrease of α-helix and β-turn and an increase of β-sheet and random coil. ITC results showed that there was a higher binding number of OA to LF than to α-LA, while both of the proteins interacted with OA through van der Waals forces and hydrogen bonds. This study provides a theoretical basis for further exploration of protein–OA complexes.

Anticancer C,N-Cycloplatinated(II) Complexes Containing Fluorinated Phosphine Ligands: Synthesis, Structural Characterization, and Biological Activity

A series of potent C,N-cycloplatinated(II) phosphine antitumor complexes containing fluorous substituents in the cyclometalated or the ancillary phosphine ligands [Pt(C-N)(PR3)Cl] or both have been synthesized and characterized. The crystal structure of [Pt(dmba){P(C6H4CF3-p)3}Cl]·2CH2Cl2 (dmba = dimethylaminomethyl)phenyl) has been established by X-ray diffraction. Values of IC50 of the new platinum complexes were calculated toward a panel of human tumor cell lines representative of ovarian (A2780 and A2780cisR) and breast cancers (T47D). Complexes containing P(C6H4CF3-p)3 as ancillary ligand (with a bulky and electronegative CF3 substituent in para position) were the most cytotoxic compounds in all the tested cancer cell lines. In some cases, the IC50 values were 16-fold smaller than that of cisplatin and 11-fold smaller than the non-fluorous analogue [Pt(dmba)(PPh3)Cl]. On the other hand, very low resistance factors (RF) in A2780cisR (cisplatin-resistant ovarian carcinoma) at 48 h were observed (RF ≈ 1) for most of the new compounds. Analysis of cell cycle was done for the three more active compounds in A2780. They arrest cell growth in G0/G1 phase in contrast to cisplatin (S phase) with a high incidence of late-stage apoptosis. They are also good cathepsin B inhibitors (an enzyme implicated in a number of cancer related events).

Location of Paclitaxel and Poly(ethylene glycol) in Triple Antitumor Complexes with Single-Walled Nanotubes According to the Molecular Docking Method

Location of Paclitaxel and Poly(ethylene glycol) in Triple Antitumor Complexes with Single-Walled Nanotubes According to the Molecular Docking Method

Synthesis, characterization and equilibrium studies of some potential antimicrobial and antitumor complexes of Cu(II), Ni(II), Zn(II) and Cd(II) ions involving 2-aminomethylbenzimidazole and glycine

The ternary complexes of Cu(II), Zn(II), Ni(II) and Cd(II) with 2-aminomethylbenzimidazole (AMBI) and glycine as a representative example of amino acids have been isolated and characterized by elemental analyses, IR, ESR, UV–vis, magnetic moment, molar conductance and 1H NMR spectra. AMBI behaves as neutral bidentate ligands with coordination through imidazole and amino group nitrogens while the glycine amino acid behaves as a monodenate anion with coordination involving the amino group and carboxylate oxygen after deprotonation. The magnetic and spectral data indicates a square planar geometry for both Cu2+ and Ni2+ complexes and a tetrahedral geometry for both Zn2+ and Cd2+ complexes. The isolated chelates have been screened for their antifungal and antibacterial activities using the disc diffusion method. A cytotoxicity of the compounds against colon (HCT116) and larynx (HEP2) cancer cells have been studied. The stability constants of ternary M-AMBI-Gly complexes were determined potentiometrically in aqueous solution at I=0.1moldm−3 NaCl.

Theoretic study of DNA base guanine and adenine and protein residues’ binding mode of the trans geometries of new antitumor non-classical platinum complexes containing pyridine and picoline ligand

The first and second substitution reactions of five activated trans platinum (II) compounds with DNA purine bases and protein residues are explored here. The free energy barriers relative to the first substitutions of these compounds are 19.5/22.9 kcal/mol (trans-[PtCl2(NH3)pyridine], (1), 16.5/14.6 kcal/mol (trans-[PtCl2(pyridine)2], (2), 18.3/18.0 kcal/mol (trans-[PtCl2(NH3)(2-picoline)], (3), 16.4/18.4 kcal/mol(trans-[PtCl2(NH3)(3-picoline)], (4), and 17.5/18.7 kcal/mol (trans-[PtCl2(NH3)(4-picoline)], (5), starting from trans-Pt-chloroaqua to trans-Pt-G/A monoadduct, while 22.5/24.8 kcal/mol(1), 18.5/23.9 kcal/mol(2), 24.3/24.9 kcal/mol(3), 23.3/24.3 kcal/mol(4), and 18.7/24.0 kcal/mol(5) are observed for trans-Pt-diaqua → trans-Pt-G/A monoadduct. In the second substitutions, reaction barriers for (G-Pt-G) head-to-head (HH)/head-to-tail (HT) formation are 17.6/16.3 kcal/mol(1), 17.7/14.8 kcal/mol(2),17.7/17.3 kcal/mol(3),18.9/18.0 kcal/mol(4), and 22.4/21.7 kcal/mol(5). Corresponding values for (G-Pt-A) HH/HT formation are 20.8/20.8(1), 24.9/22.9(2), 27.3/21.6(3), 27.5/24.6(4), and 28.1/25.4(5) kcal/mol in turns. The competition reactions reveal that for 1 and 4, methionine residue stabilizes the transition state (ΔGaq/ZPE = 13.1/12.7 kcal/mol) for platination more efficiently than purine bases and other protein residues, while cysteine residue owns the lower barrier height (11.1/13.4/11.8 kcal/mol) for 2, 3, and 5. The binding mechanism of trans platinum complexes containing planar amines, trans-[PtCl2(L)(L′)] (where L = L′ = pyridine; L = NH3, L′ = pyridine, or L′ = 2-picoline, 3-picoline, 4-picoline), with DNA and protein residues has been theoretically investigated. Our calculations reveal that the symmetric compound trans-[PtCl2(pyridine)2] is the most kinetically and thermodynamically preferred one not only for binding with guanine in the first substitution but also for binding with cysteine in the second substitution.

Improved pharmacological profile of the lipophilic antitumor dichloro-(N-dodecyl)-propanediamine-platinum(II) complex after incorporation into pegylated liposomes

Liposome encapsulation of platinum (Pt) drugs has emerged as a promising strategy to overcome their toxicity and cellular Pt resistance. The aim of the present work was to examine the impact of liposome encapsulation of a novel antitumor lipophilic Pt complex, dichloro-(N-dodecyl)-propanediamine-platinum(II) complex (DDPP), on its pharmacological profile as an antitumor agent. Biological assays included acute toxicity and histopathological evaluations, pharmacokinetics, and growth inhibition of B16-F1 tumor cells in C57Bl/6 mice. Comparison was made with cisplatin and free DDPP dissolved in castor oil. DDPP encapsulated in pegylated liposomes showed reduced acute toxicity in mice following intraperitoneal administration, compared with the free complex. Free DDPP at 5 mg Pt/kg induced histopathological alterations in the liver, in contrast to liposomal DDPP and cisplatin. Interestingly, the marked loss of body weight following the treatment of mice with cisplatin was not observed after liposomal DDPP at the same Pt dose. Liposomal DDPP was found to inhibit tumor growth significantly, when administered at 5 mg Pt/kg/day for 3 days, similar to cisplatin, but in contrast to the free complex. Pharmacokinetic studies after intraperitoneal and intravenous administrations at 5 mg Pt/kg indicated greater and more prolonged Pt levels in the plasma, liver, spleen, and kidneys from liposomal DDPP, compared with free DDPP or cisplatin. The tumor concentration of Pt increased after liposomal DDPP over the 24-h period, whereas it decreased after cisplatin. In conclusion, the encapsulation of DDPP in pegylated liposomes reduced the drug toxicity and enhanced its antitumoral activity in mice, as a result of improved drug pharmacokinetics.

Complex formation reactions and equilibrium studies of mixed ligand complexes of diaqua (1-Phenyl piperazine)(Palladium)(II) with some biologically relevant ligands.

With the purpose of the search of new antitumor metal complexes, the complex-formation reactions of [Pd(Phenpip)(H2O)2] 2+ (Phenpip = 1-Phenylpiperazine) with some selected bio-relevant ligands, containing different functional groups (amino acids, peptides, DNA constituents and dicarboxylic acids) were investigated. Stoichiometry and stability constants for the complexes formed are reported. The results show the formation of 1 : 1 complexes with amino acids and dicarboxylic acids. The effect of chelate ring size of the dicarboxylic acid complexes on their stability constants was examined. Peptides form both 1 : 1 complexes and the corresponding deprotonated amide species. Structural effects of the peptide on the amide deprotonation were investigated. DNA pyrimidinic constituents, such as uracil, uridine, thymidine and thymine, form 1 : 1 and 1 : 2 complexes, whereas purinic constituents, such as inosine, inosine 5´-monophosphate (5-IMP) form only 1 : 1 complexes. The stability constant of the complexes formed in solution were determined and the binding centers of the ligands were assigned. The concentration distribution diagrams of the complexes were evaluated

Influence of pH on the Structure and Oleic Acid Binding Ability of Bovine α-Lactalbumin

The biological function of α-lactalbumin (α-LA) depends on its conformation. α-LA can adopt a stable intermediate state induced by heating or pH change. This intermediate state associates with oleic acid (OA) to form an anti-tumor complex. The effect of temperature on the formation the complex has been studied, whereas the effect of pH on complex formation remains unresolved. The effect of pH on tryptophan residues, hydrophobic clusters and secondary structure of Ca(2+)-depleted bovine α-LA (BLA) was studied by fluorescence spectroscopy and circular dichroism. BLA was found to adopt a more flexible conformation between pH 7.0 and 9.0 with buried hydrophobic clusters. The binding ability of α-LA towards OA and the anti-tumor activity of the corresponding complex were also studied. BLA was found to bind more OA over the pH range of 7.0-9.0 and the corresponding complexes showed a higher anti-tumor activity than those complexes formed under acidic conditions. Our study indicates that alkaline pH aided the formation of the complex as well as its anti-tumor activity. We also propose a possible characteristic structure that facilitates binding of OA.

Understanding trans platinum complexes as potential antitumor drugs beyond targeting DNA

This focused review is devoted to our advances in developing trans platinum complexes as antitumoral complexes, in particular those bearing aliphatic amines. It includes some fundamental aspects of the understanding of non conventional metallodrug mechanism. The perspective is arranged by those fundamental aspects and within each section only the most successful examples have been included and also those in which clearly SAR's (structure activity relationships) have been observed.

Correction to “Differences in the Cellular Response and Signaling Pathways between Cisplatin and Monodentate Organometallic Ru(II) Antitumor Complexes Containing a Terphenyl Ligand”

Correction to “Differences in the Cellular Response and Signaling Pathways between Cisplatin and Monodentate Organometallic Ru(II) Antitumor Complexes Containing a Terphenyl Ligand”

2PT150 モルテングロビュール状態の蛋白質を利用した新規抗腫瘍複合体の作製(日本生物物理学会第50回年会(2012年度))

2PT150 モルテングロビュール状態の蛋白質を利用した新規抗腫瘍複合体の作製(日本生物物理学会第50回年会(2012年度))

Pharmacokinetic Study of Di-Phenyl-Di-(2,4-Difluobenzohydroxamato)Tin(IV): Novel Metal-Based Complex with Promising Antitumor Potential

Di-phenyl-di-(2,4-difluobenzohydroxamato)tin(IV)(DPDFT), a new metal-based arylhydroxamate antitumor complex, showed high in vivo and in vitro antitumor activity with relative low toxicity, but no data was reported regarding its pharmacokinetics and dependent toxicity. In this paper, a rapid, sensitive, and reproducible HPLC method in vivo using Diamonsil ODS column with a mixture of methanol and phosphoric acid in water (30 : 70, V/V, pH 3.0) as mobile phase was developed and validated for the determination of DPDFT. The plasma was deproteinized with methanol that contained acetanilide as the internal standard (I.S.). The photodiode array detector was set at a wavelength of 228 nm at room temperature and a linear curve over the concentration range 0.1~25 μg·mL−1 (r = 0.9993) was obtained. The method was used to determine the concentration-time profiles for DPDFT in the plasma after single intravenous administration with doses of 5, 10, 15 mg·kg−1 to rats. The pharmacokinetics parameter calculations and modeling were carried out using the 3p97 software. The results showed that the concentration-time curves of DPDFT in rat plasma could be fitted to two-compartment model.

Novel C,N-chelate rhodium(iii) and iridium(iii) antitumor complexes incorporating a lipophilic steroidal conjugate and their interaction with DNA

The novel steroidal conjugates [M(η(5)-C(5)Me(5))Cl(LEV-ppy)] (M = Rh (1) and Ir (2)) bearing the lipophilic levonorgestrel group 17-α-[2-phenylpyridyl-4-ethynyl]-19-nortestosterone (LEV-ppy), where the chelating ligand is N and C-bound, have been prepared and characterized. Both compounds are more active than cisplatin (about 6-fold) in T47D (breast cancer) at 48 h incubation time. On the other hand, very low resistance factors (RF) of 1 and 2 in A2780cisR (cisplatin-resistant ovarian carcinoma) at 48 h were observed (RF = 0.9 and 1.1, respectively). The iridium steroidal compound 2 is twice as active as the non-steroidal analogue 2', whose promising anticancer activity has recently been reported by Sadler. Theoretical DFT calculations on complexes 1 and 2 at the B3LYP-D/def2-TZVP-ecp level of theory show that the strongest bond to the metal atom is the η(5)-interaction to the Cp* ligand and that both of them feature a rather strong metal-chlorine bond. The new steroidal conjugates 1 and 2 are able to bind to DNA according to Hoechst 33258 displacement experiments and ESI-TOF MS spectrometry studies. Complexes 1 and 2 are also cathepsin B inhibitors, an enzyme implicated in a number of cancer related events.

Inhibition of Cytochrome P450 3A1/2 by Organotin Compound Di-N-Butyl-(4-chlorobenzohydroxamato)tin (IV) Chloride in Rat Liver and BRL Cells

Organotin compounds are high toxiferous chemicals and ubiquitous in our environment, which also have high biological activities. Di-n-butyl-(4-chlorobenzohydroxamato)tin (IV) chloride (DBDCT) represents a new paradigm for tin-based antitumor complexes with high activity. The inhibitory effect of DBDCT on cytochrome P450 3A(CYP3A) was studied in this article. The adult male SD rats were randomly divided into five groups with six in each and treated separately with saline, lipopolysaccharide (LPS, 5mg/kg), DBDCT (1.25, 2.5 and 5.0mg/kg, respectively) intraperitoneally for 2 days after induced with dexamethasone (DEX) at a dose of 100mg/kg for 4 days. The cytochrome P450 (CYP450) content was assayed by the method of Omura and Santa after the protein concentration detected by BCA assay kit. The activity of CYP3A was determined by the method of Nash. Western blot analysis was used to detect the expression of CYP3A1/2 at protein level in rat liver microsomes and Buffalo Rat Liver (BRL) cells. The results demonstrated that the activity of CYP450 and CYP3A were inhibited by exposure to DBDCT in rat liver compared with that of the blank control group. The expression of CYP3A1 and CYP3A2 proteins in rats treated with 1.25, 2.5 and 5.0mg/kg DBDCT were down-regulated respectively by 22.8% (p<0.01),24.3%(p<0.01), 58.4%(p<0.001) and 37.6%(p<0.001), 41.4%(p<0.001), 49.2%(p<0.001), than that of the blank control group. Immunoblot analysis of protein from BRL cell lysate demonstrated that the expression of CYP3A1 and CYP3A2 protein reduced separately by 11.4% (p<0.05), 34.2%(p<0.001), 45.5%(p<0.001), 64.9%(p<0.001), 78.8%(p<0.001) and 12.8% (p<0.05), 9.9% (p<0.05), 28.0% (p<0.001), 24.9% (p<0.001), 34.2% (p<0.001) after treated with DBDCT at a dose of 1µmol/L (for 24, 36, 48h) and 2µmol/L(for 24 and 48h). The activity of CYP3A in rat liver microsomes was decreased remarkably compared with that of the blank control group. Immunoblot analysis showed that the expression of CYP3A1/2 was inhibited significantly by DBDCT in rat liver and BRL cells.

A DFT STUDY ON THE HYDROLYSIS MECHANISM OF THE NAMI-A-TYPE ANTITUMOR COMPLEX (HL)[trans-RUCl4L(dmso-S)](L=4-amino-1,2,4-triazole)

The hydrolysis process of Ru (III) complex (HL)[trans- RuCl4L(dmso-S)] (L=4-amino-1,2,4-triazole) (1), a potential antitumor complex similar to the well-known antitumor agent (ImH)[trans- RuCl4(dmso-S)(Im)](NAMI-A), was investigated using density functional theory (DFT) with the conductor-like polarizable continuum model (CPCM). The structural characteristics and the detailed energy profiles for the hydrolysis processes of this complex were obtained. For the first hydrolysis step, complex 1 with 4-amino-1,2,4-triazole ligand shows much faster aquation than NAMI-A with imidazole ligand and complex 2 with 4H-1,2,4-triazole ligand, and such a calculated result is in good agreement with the experimental one. For the second hydrolysis step, the formation of cis-diaqua products is found to be thermodynamically preferred over the trans isomers. In addition, on the basis of the analysis of electronic characteristics of species in the hydrolysis process, the trend in abilities (A) of hydrolysis products attacked nucleophilicly by pertinent biomolecules is revealed. These theoretical results will help in understanding the action mechanism of this potential Ru (III) drug with pertinent biomolecular targets.

Hydrolysis Mechanism of the NAMI-A-type Antitumor Complex (HL)[trans-RuCl4L(dmso-S)] (L=1-methyl-1,2,4-triazole)

The hydrolysis process of Ru(III) complex (HL)[trans-RuCl4L(dmso-S)] (L=1-methyl-1,2,4-triazole and dmso-S=S-dimethyl sulfoxide) (1), a potential antitumor complex similar to the well-known antitumor agent (Him)[trans-RuCl4(dmso-S)(im)] (NAMI-A, im=imidazole), was investigated using density functional theory combined with the conductor-like polarizable continuum model approach. The structural characteristics and the detailed energy profiles for the hydrolysis processes of this complex were obtained. For the first hydrolysis step, complex 1 has slightly higher barrier energies than the reported anticancer drug NAMI-A, and the result is in accordance with the experimental evidence indicating larger half-life for complex 1. For the second hydrolysis step, the formation of cis-diaqua species is thermodynamic preferred to that of trans isomers. In addition, on the basis of the analysis of electronic characteristics of species in the hydrolysis process, the trend in nucleophilic attack abilities of hydrolysis products by pertinent biomolecules is revealed and predicted.