Damian P. Buck

Solubilisation and cytotoxicity of albendazole encapsulated in cucurbit[n]uril

The aqueous solubilities of albendazole encapsulated in cucurbit[6, 7 and 8]urils (Q[6], Q[7] and Q[8]) have been determined by (1)H NMR spectroscopy, and the effect of encapsulation on their cytotoxicities evaluated. Encapsulation in Q[6] and Q[7] increased the aqueous solubility of albendazole by 2000-fold, from 3 microM to 6 mM at pH 6.6, while Q[8]-encapsulation increased the solubility to over 2 mM. Encapsulation in Q[7] and Q[8] induced significant upfield shifts for the albendazole propyl and benzimidazole resonances, compared to those observed for Q[6]-binding and what would normally be expected for the respective functional groups. The upfield shifts indicate that the albendazole propyl and benzimidazole protons are located within the Q[7] and Q[8] cavity upon encapsulation. Alternatively, encapsulation in Q[6] only induced a large upfield shift for the albendazole carbamate methyl resonance, indicating that the drug associates with Q[6] at its portals, with only the carbamate group within the cavity. Simple molecular models based on the observed relative changes in chemical shift could be constructed that were consistent with the conclusions from the NMR experiments. Cytotoxicity assays against human colorectal cells (HT-29), human ovarian cancer cells (1A9) and the human T-cell acute lymphoblastic leukaemia cells (CEM) indicated that encapsulation in Q[7] did not significantly reduce the in vitro anti-cancer activity of albendazole.

Cucurbituril binding of trans-[{PtCl(NH3)2}2(µ-NH2(CH2)8NH2)]2+ and the effect on the reaction with cysteine

The effect of encapsulation by cucurbiturils Q[7] and Q[8] on the rate of reaction of the anti-cancer dinuclear platinum complex trans-[{PtCl(NH3)2}2(micro-NH2(CH2)8NH2)]2+ with the model biological nucleophiles glutathione and cysteine has been examined by NMR spectroscopy. It was expected that the octamethylene linking chain would fold inside the cucurbituril host and hence position the reactive platinum centres close to the cucurbituril portals, and thereby, confer resistance to degradation by biological nucleophiles. The upfield shifts of the resonances from the methylene protons in the linking ligand observed in 1H NMR spectra of the platinum complex upon addition of either Q[7] or Q[8] indicate that the cucurbituril is positioned over the linking ligand, with the Pt(II) centres projecting out of the portal. Furthermore, the relative changes in chemical shift of the methylene resonances suggest that the octamethylene linking chain folds within the cucurbituril cavity, particularly in Q[8]. Simple molecular models, based on the observed relative changes in chemical shift, could be constructed that were consistent with the proposed folding of the linking ligand within the cucurbituril cavity. Encapsulation by Q[7] was found to reduce the rate of reaction of the platinum complex with glutathione. Encapsulation by Q[7] and Q[8] was also found to reduce the rate of reaction of the platinum complex with cysteine, with Q[8] slowing the reaction to a greater extent than Q[7], consistent with the inferred encapsulation geometries. Encapsulation of dinuclear platinum complexes within the cucurbituril cavity may provide a novel way of reducing the reactivity and degradation of these promising chemotherapeutic agents with blood plasma proteins.

Binding of a dinuclear ruthenium(II) complex to the TAR region of the HIV-AIDS viral RNA

Molecular modelling has identified a new RNA conformational feature created by the insertion of bulge residues into duplex regions that may act as a recognition site for small molecule binding, in particular for inert dinuclear ruthenium complexes.

Binding of a Flexibly-linked Dinuclear Ruthenium(II) Complex to Adenine-bulged DNA Duplexes

Using 1H NMR spectroscopy and molecular modelling, the DNA binding of a chiral dinuclear ruthenium(ii) complex {Δ,Δ-[{Ru(phen)2}2(μ-bb7)]4+; phen = 1,10-phenanthroline, bb7 = 1,7-bis[4(4′-methyl-2,2′-bipyridyl)]-heptane} involving a bridging ligand containing a flexible aliphatic chain has been studied. The binding of the ruthenium(ii) complex was examined with the non-self-complementary duplexes d(CCGAGAATCGGCC):d(GGCCGATTCCGG) (containing a single adenine bulge: designated SB) and d(CCGAGCCGTGCC):d(GGCACGAGCCGG) (containing two adenine bulge sites separated by two base-pairs: designated DB). The NMR data indicated that the ruthenium(ii) complex bound at the bulge site of SB, with one ruthenium centre located at the bulge site with the second metal centre binding with lower affinity and selectivity in the duplex region adjacent to the bulge site. Less specific binding is inferred from chemical shift changes of nucleotide protons two to five base pairs from the single adenine bulge. The ruthenium(ii) complex selectively bound the DB duplex with one metal centre located at each bulge site. The NMR results also suggested that the metal complex binding induced greater changes to the structure of the SB duplex, compared with the DB duplex. Modelling indicates the bridging ligand allowed each ruthenium(ii) metal centre to bind one adenine bulge of the doubly-bulged duplex without disrupting the DNA structure, using the additional torsional flexibility conferred by the aliphatic bridging ligand. However, the second ruthenium(ii) metal centre is not able to bind in the minor groove of the singly-bulged duplex without disrupting the structure, as the metal centre is too bulky. The results of this study suggest dinuclear ruthenium(ii) complexes have considerable potential as probes for DNA and RNA sequences that contain two bulge sites separated by a small number of base-pairs.

Electrochemical reduction of nitrotriazoles in aqueous media as an approach to the synthesis of new green energetic materials

The synthesis of new azo and azoxy compoundsvia electrochemical reduction of nitrotriazoles has been investigated in aqueous media, using nitrotriazolone (NTO) and nitrotriazole (NTr) as representative substrates. Reduction of NTO produces mainly solid azoxytriazolone (AZTO), with azotriazolone (azoTO) and aminotriazolone (ATO) as minor products, while 3-hydroxylaminotriazole is the major product formed from NTr. AZTO and azoTO are of interest as new green high-nitrogen compounds for use as insensitive high explosives (IHE). The effect of varying reaction conditions such as pH and substrate concentration has been evaluated, and a mechanism is proposed accounting for the experimental observations. In particular, the ratio of azoxy to azo in the solid product is influenced by pH and temperature, and the minor product ATO is formed not via direct reduction of NTO but via a novel thermal disproportionation reaction of the hydrazotriazolone intermediate. Conditions of high substrate concentration and low cell temperature maximise the azoxy yield and minimise the formation of minor products. Results indicate that this green electrosynthetic approach may be generally useful for the synthesis of new azoxy and azo triazoles from suitable substrates.

NMR Studies of Metallointercalator–DNA Interactions

The study of the interaction between inert transition metal complexes and nucleic acids has developed from the early work of Dwyer [1], Lippard [2], Norden [3] and Barton [4] to the point that it is now a central theme in bio-inorganic chemistry. While there has been considerable interest in metal complexes that bind nucleic acids, the interaction of metallointercalators with DNA and RNA has received the most attention [5, 6]. Square-planar platinum(II) complexes have demonstrated significant anticancer activity [7], and octahedral ruthenium(II) and rhodium(III) complexes have been used as probes of nucleic acid structure and as a means to study electron transfer reactions mediated by the heteroaromatic bases [5, 6]. While a range of techniques is available to study the nucleic acid binding of metal complexes, NMR spectroscopy (particularly 1H NMR) has proven to be the most useful. NMR spectroscopy can provide a detailed, atom level resolution, picture of the metal complex binding, and if the quality of the data is sufficient, a threedimensional structure of the metal complex bound to the oligonucleotide can be determined. The strategies used to assign the 1H NMR spectrum of an oligonucleotide [8–10], the extension of these methods to study the interaction of metal complexes with DNA and the use of molecular modelling will be presented in this chapter.