Comprehensive exploration of the anticancer activities of procaine and its binding with calf thymus DNA: a multi spectroscopic and molecular modelling study

Abstract

Procaine is an anesthetic drug commonly administrated topically or intravenously for use in local anesthesia. Promisingly, some anticancer activities of procaine have also been reported. Therefore, the mechanism of interaction between anesthetic drug procaine with ct-DNA was determined collectively by means of various spectroscopic and molecular docking methods. Minor groove 1 : 1 binding of procaine to the ct-DNA was evidenced from absorption spectroscopy, fluorescence quenching, DNA melting, competitive binding measurements with EB and DAPI dyes, viscosity and CD spectroscopy together with molecular docking simulations and DFT calculations. Molecular docking on five different B-DNA structures (taken from the Protein Data Bank) shows that procaine binds in the AT rich region of all five B-DNA structures. Thermodynamic parameters, evaluated using van't Hoff's isotherm, shown that the interaction was feasible and the binding forces involved were hydrophobic as well as hydrogen bonding which were, further, confirmed by molecular docking. The frontier molecular orbitals (HOMO and LUMO) of procaine and DNA bases have been calculated by DFT method and the chemical potential (m), chemical hardness (h) and fraction number of electrons (DN) from procaine to DNA bases were evaluated, which have shown that procaine acts as an electron donor to the DNA bases. Simultaneously, anticancer activities of procaine alone and in combination with doxorubicin were observed on the MCF-7 breast cancer cell line. The results showed that the combined treatment with both procaine and doxorubicin enhanced the cytotoxic and apoptotic inducing potential of doxorubicin.

Design, synthesis, and DNA interaction studies of furo-imidazo[3.3.3]propellane derivatives: Potential anticancer agents

“…In the same time and as shown in Fig. 6A and 6B, the three propellane derivatives 5c, 5d and 5f were able to replace the minor groove binder Hoechst 33258 dye from CT-DNA in a dose dependent manner with EC 50 ′s 9, 19 and 34 μM respectively ( [46,47] or DAPI [56][57][58] dye from the minor groove of the DNA helix, resulting in decreased fluorescence intensity of Hoechst-DNA or DAPI-DNA system. These experiments suggest the binding mode of these new propellanes to be groove binding rather than intercalation, the observation that requires several future experiments to be confirmed.…”

“…In this experiment, we tested the effect of the new propellane derivatives on the ability of EB or TO dye to intercalate with CT-DNA and DAPI or Hoechst 33258 dye to bind the CT-DNA groove. DAPI has been reported to bind the DNA minor groove (in the case of A-T rich DNA) in addition to its ability to intercalate with the DNA (in the case of G-C rich DNA) [56,58]. The calf thymus DNA used in this study is 41.9 mol % G-C and 58.1 mol % A-T, which makes DAPI to act mostly as a minor groove binder in this experiment.…”

“…Replacing such a dye by a ligand may suggest the ligand binding site. Thus, intercalator ligands are expected to replace an intercalator dye, such as ethidium bromide (EB) or thiazole orange (TO), while a groove binder is expected to replace a groove binding dye, such as DAPI or Hoechst 33258 [46,47,52,56,57]. In this experiment, we tested the effect of the new propellane derivatives on the ability of EB or TO dye to intercalate with CT-DNA and DAPI or Hoechst 33258 dye to bind the CT-DNA groove.…”

Fluorescent delivery vehicle containing cobalt oxide–umbelliferone nanoconjugate: DNA/protein interaction studies and anticancer activity on MF7 cancer cell line

“…HSA and DNA binding studies were carried out using UV-visible, uorescence quenching and circular dichroism methods and the detailed experimental procedures for these studies have been described elsewhere. 20,[23][24][25][26][27]…”

“…It is considered that the binding site of the complex is somewhere at the interfacial region of the intercalating site and groove region. To conrm our hypothesis we have performed dye displacement assays using EB which is a classical intercalator and DAPI which is known to bind at minor groove under our experimental conditions 26,27 and the results are shown in Fig. 6.…”