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N Farrell

Publications and source records attributed to N Farrell.

50 records · Page 3Linked to original sources
PubMed1994

Sequence specificity of DNA-DNA interstrand cross-link formation by cisplatin and dinuclear platinum complexes.

The sequence specificity of interstrand cross-links induced in DNA by mononuclear and dinuclear platinum complexes in a 49-base-pair DNA duplex has been determined directly. This new assay takes advantage of the fact that 3'-->5' exonuclease digestion of randomly platinated DNA produces a pool of fragments of different lengths. This treatment allows identification of the spectrum of adducts impeding the exonuclease scission. Interstrand cross-linked adducts produce fragments that may remain complementary in the proximity of the binding site. As a result, these fragments may act as primer templates for extension upon subsequent treatment with a DNA polymerase. This extension increases the size of the oligonucleotide fragments, which may be evidenced by a more slowly migrating band on a sequencing gel. Concomitantly, the original band corresponding to the digested cross-link decreases in intensity. Therefore, comparison of a sequencing gel after digestion only and after the "digestion-extension" treatment should show the disappearance, or diminished band intensity, of only those fragments with interstrand cross-links. This approach was applied to the analysis of DNA interstrand cross-links formed by cis-[PtCl2(NH3)2] (cis-DDP) and [(trans-PtCl(NH3)2)2H2N(CH2)4NH2]Cl2. Cis-DDP was confirmed to form interstrand cross-links at d(GC) sequences but, interestingly, interstrand cross-links predominated in a sequence GCGG, with possible 1,3-intrastrand but no 1,2-intrastrand cross-links forming. The dinuclear compound formed 1,2, 1,3, and 1,4 DNA interstrand cross-links between guanines on opposite strands. In 1,3 and 1,4 cross-links, the guanines are separated by one and two base pairs, respectively, whereas a 1,2 cross-link is formed from guanines on neighboring base pairs.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗
PubMed1994

Radiosensitizing and toxic properties of quinoline and nitroquinoline complexes of platinum [PtCl2(NH3)quinoline].

The assessment of drugs designed to be useful in the eradication of hypoxic (resistant) cells involves comparison of hypoxic and aerobic radiosensitization, cytotoxicities, as well as DNA binding and reduction potentials. Three pairs of isomers of quinoline complexes [amino dichloro quinoline platinum (II)] were studied in this context. For the cis 5- and 6-nitroquinoline complexes, the DNA binding and toxicity were higher with the 5-substituted ligand. Reduction potentials were similar (-260 and -280 mV). No selectivity for hypoxic toxicity was observed, but radiosensitizing ability by both complexes was greater in hypoxic (than oxic) Chinese hamster ovary (CHO) cells. The trans 5-nitroquinoline complex produced better sensitization in hypoxia than its cis isomer [enhancement ratio (ER) 1.7 at 10 microM versus 40 microM for cis]. However, this was accompanied by some aerobic sensitization. The trans isomer of the (unsubstituted) quinoline complex was considerably more toxic than its cis isomer. Neither showed selectivity for hypoxia, either as radiosensitizers or as cytotoxins, which may be attributable to the lack of a reducible (nitro) function. Four quinoline complexes showed high activity in cisplatin-resistant L1210 cells, with the lowest resistance factor being for the trans quinoline complex. Results suggest that trans complexes with one aromatic ring may have activity different from the cis geometry, which should be exploited with respect to cisplatin resistance and cross-resistance with radiation.

Animals↗
PubMed1993

Homodinuclear (Pt,Pt) and heterodinuclear (Ru,Pt) metal compounds as DNA-protein cross-linking agents: potential suicide DNA lesions.

Homodinuclear (Pt,Pt) and heterodinuclear (Ru,Pt) metal compounds having the generalized formula M(a)NH2(CH)4NH2M(b) are shown to form specific DNA lesions which can efficiently cross-link proteins to DNA. In this study, the homodinuclear case is represented by M(a) = M(b) = [cis-Pt(Cl2)-(NH3)] and the heterodinuclear case is represented by M(a) = [cis-RuCl2(DMSO)3] and M(b) = [cis-PtCl2(NH3)]. Native and denaturing polyacrylamide gel electrophoresis was used to show the formation of ternary coordination complexes between the metal-treated 49-bp DNA fragment and the Escherichia coli UvrA and UvrB DNA repair proteins. Treatment with proteinase K results in loss of the DNA-protein cross-links. DNA-protein cross-links formed between UvrA and DNA previously modified with the dinuclear metal compounds are reversible with the reducing agent beta-mercaptoethanol. The DNA lesion responsible for efficient DNA-protein cross-linking is most probably a DNA-DNA interstrand cross-link in which each metal atom is coordinated with one strand of the DNA helix. The formation of DNA repair protein associated DNA cross-links, potential "suicide adducts", suggests a novel action mechanism for these anticancer compounds. In addition, these dinuclear metal compounds should be very useful agents for the investigation of a wide range of protein-DNA interactions.

Adenosine Triphosphatases↗
PubMed1993

Ligand effects on platinum binding to DNA. A comparison of DNA binding properties for cis- and trans-[PtCl2(amine)2] (amine = NH3, pyridine).

The DNA binding properties of cis- and trans-[PtCl2(pyridine)2] have been examined and compared with their NH3 analogs, cis- and trans-DDP. The presence of a planar ligand reduces the rates of DNA binding but does not greatly affect the overall conformation of CT DNA, as measured by circular dichroism spectroscopy. The sequence specificity of trans-[PtCl2(py)2] includes alternating purine-pyrimidine sequences. The sequence specificity is further different between the two pyridine isomers, and the steric effects of two cis-pyridine groups are demonstrated by the appearance of relatively few binding sites in the 49-bp duplex. The effects of the pyridine ligand are further manifested by a greatly enhanced DNA-DNA interstrand cross-linking efficiency for the trans isomer, with a cross-link per adduct frequency of between 0.14 and 0.23, depending on the rb of the sample. The unwinding of closed circular pUC19 DNA by trans-[PtCl2(pyridine)2] is also more efficient than that by either DDP isomer, with an unwinding angle calculated at phi = 17 degrees (compare cis-DDP with phi = 13 degrees and trans-DDP with phi = 9-10 degrees). In contrast, little unwinding is induced by cis-[PtCl2(pyridine)2], with phi = 4 degrees. These results in particular invert the standard cis/trans structure-activity relationships observed previously for [PtCl2(NH3)2]. The results are discussed with respect to the previously demonstrated effect of activation of the trans-platinum geometry using sterically hindered ligands.

Base Sequence↗
PubMed1993

Nonclassical platinum antitumor agents: perspectives for design and development of new drugs complementary to cisplatin.

Studies over the last few years have shown that the range of platinum complexes with useful cytotoxicity and antitumor activity is not strictly limited to structural analogs of cisplatin. In general, we can expect that cells will process structurally different species in different manners. The metabolic chemistry and DNA binding will be altered in in comparison to the cis-[PtX2 (amine)2] class. This point is of particular importance because any altered pattern of antitumor activity of structural analogs of cisplatin is likely to be due to unpredictable pharmacokinetic, rather than truly mechanistic, factors. The fact that discrete cisplatin-DNA adducts vary in their biological activity further supports the hypothesis that complexes structurally dissimilar to cisplatin may produce biological activity complementary to the parent drugs. The mechanism of action of nonclassical complexes is different from that of cisplatin and its analogs. Their pattern of antitumor activity is also altered with respect to cisplatin--thus, not all platinum-containing drugs need necessarily be similar in their clinical profile to cisplatin. Note that both the dinuclear bis(platinum) complexes and the trans complexes give their own distinct patterns of tumor specificity--different from cisplatin and each other (see Tables 1 and 3). New cytotoxic mechanisms for platinum complexes may also be placed in context with cisplatin resistance. Modes of overcoming cisplatin resistance may reside at the various levels of uptake, interaction with "detoxifying" intracellular thiols, and DNA repair. Likewise complexes with novel mechanisms of action may circumvent resistance by more than one unique route. Indeed, the three major routes to resistance are all affected to varying degrees by the complexes outlined above. From the discovery of cisplatin, the development of analogs has essentially been an empirical exercise. Because of their similar mechanism of action, much comparison has been made between platinum complexes and the classic alkylating agents. Yet the alkylating agents represent a good example where a number of structurally distinct drugs with different anticancer activities are clinically available. This desirable feature may be achieved for platinum complexes by emphasis on complexes structurally dissimilar to the presently used agents. The dinuclear bis(platinum) complexes and mononuclear complexes in the trans geometry are of special interest. Comparison of common features and differences between different classes may point to guidelines for the rational design of complexes with a different spectrum of clinical antitumor activity to cisplatin and activity against cisplatin-resistant tumors.

Antineoplastic Agents↗
PubMed1992

Activation of the trans geometry in platinum antitumor complexes: a survey of the cytotoxicity of trans complexes containing planar ligands in murine L1210 and human tumor panels and studies on their mechanism of action.

The cytotoxicity of transplatinum complexes of structural formula trans-[PtCl2(L)(L')] [L = L' = pyridine or thiazole, or L = quinoline (R' = methyl; R" = methyl, phenyl, or CH2phenyl) and L' = R'R"SO] has been studied in murine L1210 and human tumor cell lines. The results confirm previous observations that use of a sterically hindered planar ligand greatly enhances cytotoxicity, in comparison to trans-[PtCl2(NH3)2], such that in some cases cytotoxicity equivalent to that of the clinically used agent cisplatin [cis-[PtCl2(NH3)2]] is obtained. Results from both the panel of human ovarian carcinoma cell lines and the National Cancer Institute screening panel confirm a different pattern of cytotoxicity, with respect to cisplatin. The new trans-platinum complexes are also non-cross-resistant with cisplatin in both murine and human (human ovarian carcinoma panel) tumor cell lines. Preliminary mechanistic studies using both cis- and trans-[PtCl2(pyridine)2] in L1210 cells have been carried out, to delineate the reasons for both the dramatically enhanced cytotoxicity and the lack of cross-resistance with the clinically used agents. Intracellular uptake is enhanced for pyridine relative to ammine (NH3) complexes. The pyridine complexes also inhibit DNA synthesis, implying a role for DNA binding in their mechanism of action. Binding of the pyridine complexes to calf thymus DNA is, however, significantly less than for the analogous ammine complexes. The presence of trans-pyridine ligands results in steric hindrance, which retards the rate of reaction of trans-[PtCl2(pyridine)2], relative to trans[PtCl2(NH3)2], with other important biomolecules such as glutathione. The results point to a potential new class of platinum antitumor complexes acting by a new mechanism and with activity complementary to agents such as cisplatin.

Animals↗
PubMed1992

B----Z DNA conformational changes induced by a family of dinuclear bis(platinum) complexes.

The reactions of bis(platinum) complexes of general formula [(PtClm(NH3)3-m)2(NH2(CH2)nNH2)]2(2-m)+ were studied with poly(dG-dC).poly(dG-dC), poly(dG-m5dC).poly(dG-m5dC) and poly(dG).poly(dC). When m = 0 (Complexes II, n = 2,4) the complexes are saturated 4+ cations capable only of electrostatic interactions with the polynucleotide. Where m = 1 the complexes contain two monodentate platinum coordination spheres with the chloride trans to the diamine bridge (Complexes I, n = 2,4, 1,1/t,t). Complexes I give CD spectra characteristic of a 'Z-like' conformation upon reaction with poly(dG-dC).poly(dG-dC) and poly(dG-m5dC).poly(dG-m5dC) but not poly(dG).poly(dC). The B----Z transition appears independent of interplatinum diamine chain length. As little as 1 bis(platinum) complex per 25-30 base pairs is sufficient to observe the Z-like spectrum. Covalent binding is however not a prerequisite for Z-DNA formation because the polyvalent cations II are also very effective in inducing the B----Z transition in either poly(dG-dC).poly(dG-dC) or poly (dG-m5dC).poly(dG-m5dC). In these cases, the concentrations of II required are significantly lower than analogous monomeric agents such as [Co(NH3)6]3+. The possible biological consequences of the Z-DNA induction by bis(platinum) complexes are discussed.

Circular Dichroism↗
PubMed1991

Shikimic acid complexes of platinum. Preparation, reactivity, and antitumor activity of (R,R-1,2-diaminocyclohexane) bis(shikimato) platinum(II). Evidence for a novel rearrangement involving platinum-carbon bond formation.

The complex (R,R-1,2-diaminocyclohexane)bis(shikimato)platinum(II) (shikimato = the anion of 3R,4S,5R-trihydroxy-1-cyclohexene-1-carboxylic acid), I, has been synthesized and purified by high performance liquid chromatography (HPLC). The complex is only moderately stable in aqueous solution. Its major hydrolysis product, also purified by HPLC, is proposed to be a unique complex type in which a single shikimate group is coordinated through both the carboxylate oxygen and the C(2) vinylic carbon of the shikimate moiety [Pt(R,R-dach)(O,C-shikimato)], II. In vitro, complex I is active against L1210 leukemia and against an L1210 cell line with acquired resistance to cisplatin. In vivo, the complex is active against L1210, P388, and B16 melanoma; this activity is highly schedule-dependent. Complex II is also active against L1210 leukemia.

Animals↗
PubMed1990

Comparison of chemical reactivity, cytotoxicity, interstrand cross-linking and DNA sequence specificity of bis(platinum) complexes containing monodentate or bidentate coordination spheres with their monomeric analogues.

The properties of a new bis(platinum) complex containing two monodentate coordination spheres, [(trans-PtCl(NH3)2)2H2N(CH2)4NH2]Cl2 (1,1/t,t), are reported. Comparison is made with respect to chemical reactivity, in vitro biological activity in murine and tumor cells, DNA conformational changes, cross-linking efficiency, and sequence specificity between this complex and the previously reported complex containing two bidentate platinum atoms, [(Pt(mal)(NH3))2H2N(CH2)4NH2] (2,2/c,c), as well as with their respective monomeric analogues, [PtCl(dien)]Cl and cis-[PtCl2(NH3)2](cis-DDP). While both bis(platinum) complexes are active against cis-DDP-resistant cells, the monodentate bis(platinum) complex (1,1/t,t) has a lower resistance factor than the complex with bidentate coordination spheres (2,2/c,c). More importantly, this property is repeated in a human ovarian carcinoma cell line. DNA-binding studies show that DNA interstrand cross-linking is more efficient for the 1,1/t,t complex. DNA sequencing studies employing the exonuclease activity of T4-polymerase demonstrate that there are a variety of binding sites; some are common to all complexes and some common to both bis(platinum) complexes, while the monodentate 1,1/t,t species also reacts at unique sites, not attacked by any of the other complexes studied. The circular dichroism of CT DNA modified by the 1,1/t,t complex is also unique and is not seen for any of the other agents.

Animals↗
PubMed1990

Cytotoxicity and antitumor activity of bis(platinum) complexes. A novel class of platinum complexes active in cell lines resistant to both cisplatin and 1,2-diaminocyclohexane complexes.

The in vitro cytotoxicity and in vivo antitumor activity of bis(platinum) complexes of general formula [(PtX2-(L))2H2N(CH2)nNH2] (L = NH3, X = Cl or X2 = malonato or where L = py, X = Cl) is reported. Chloride complexes [(PtCl2(NH3]2H2N(CH2)nNH2] may exist as three possible isomers: those containing both coordination units in the cis configuration (2,2/c,c), both coordination units in the trans configuration (2,2/t,t), and the mixed cis,trans species (2,2/c,t), whose synthesis is reported here. The preparation of further complexes with sterically hindered diamine backbones, such as 2,5-dimethyl-2,5-hexanediamine, is exemplified. The biological activity of all complexes were compared. The 2,2/c,c complexes are particularly active in tissue culture in cells resistant both to cisplatin and its 1,2-diaminocyclohexane (dach) analogue. The inhibition of DNA synthesis in L1210/0 cells by the 2,2/c,c complexes is equivalent to that of cisplatin. The presence of at least one cis-[Pt(amine)2] unit appears necessary for activity in cell lines sensitive to cisplatin.

Animals↗
PubMed1987

Reactions of oxobis(acetylacetonato)-vanadium (IV) with some purine and pyrimidine bases and nucleosides.

A study of the reactions of purine and pyrimidine bases has been initiated with the V(IV) complex [V(O)(acac)2]. Two distinct modes of reaction have been recognized depending on the nature of the base. Simple adduct formation occurs when the base contains a sufficiently basic lone pair, but when bases contain acidic protons bound to nitrogen (inosine, thymine), cleavage of one acetylacetonate ligand occurs to give potentially polymeric species. This behavior is in contrast to that of the inert [Co(NO2)2(acac)2]-ion.

Nucleosides↗
PubMed1985

The effect of tityustoxin and ruthenium red on the release of acetylcholine from slices of cortex of rat brain.

The effect of tityustoxin and ruthenium red on the release of acetylcholine from slices of cortex from the brain of the rat was studied. The results showed that ruthenium red stimulated the elicited release of acetylcholine (ACh) by tityustoxin. The effect was dependent on the incubation time and the concentration of ruthenium red. The stimulation was dependent on calcium and was blocked by the presence of tetrodotoxin and ethyleneglycol-bis(beta aminoethyl ether)N,N'-tetraacetic acid (EGTA). It is suggested that the inhibition of buffering by calcium in mitochondria, by ruthenium red increased the level of ionized calcium in the symaptoplasm, causing an increase in the release of acetylcholine.

Acetylcholine↗