In-Silico and In-Vitro Evaluation of Annona Muricata Leaf Extract Conjugated with Doxorubicin Via Liposomal Drug Delivery System against (MDA-MB-231) Triple Negative Breast Cancer Cell Line

This study aims on in-silico and in-vitro evaluation of breast cancer activity. The in-silico study focused against the P-glycoprotein, which hammers the therapeutic efficacy of various chemotherapeutic agents because of P-gp efflux mechanism. The present study involves in the collection of phytoconstituents of Annona muricata leaf and evaluated against the selected targets (MDR 1, ABCB1, AP2, Cav-1, Transferrin). In-silico study involves in the analysis of drug-likeness, ADMET analysis and molecular docking study. ➢ The drug-likeness study results reveals that almost all the Phytoconstituents of Annona muricata were not suitable for oral administration due to poor bioavailability and molecular weight more than 500 Daltons and all the ligands violates the two or more violations in Lipinski rule of five. ➢ ADMET analysis of the Phytoconstituents shows the significant results. Among the compounds Annocatalin, Annomuricin A, Annomuricin B, Annomuricin C, Annomuricin-D-one, Annomuricin E, Annomutacin, Annopentocin A, Annopentocin B, Annopentocin C, Iso-Annonacin, Muricatocin B and Muricatocin C exhibits the good ADMET profile. ➢ The present study shows that DOX can deliver into cancer cell by the inhibition of P-gp efflux by phytoconstituents present in Annona muricata. So, the therapeutic efficacy of DOX can be enhanced. ➢ Molecular docking of phytoconstituents reveals that Annopentocin A shows significant binding energy of (-9.2 kcal mol−1) with MDR 1 protein, Annocatalin shows significant binding energy of (-10.5 kcal mol−1), (-10 kcal mol−1) and (-7.6 kcal mol−1) with ABCB1, AP2 and CAV-1 protein and Annomuricin B shows significant binding energy of (-9.8 kcal mol−1) with Transferrin. ➢ The Physiochemical characterization of AM-DOX liposome shows the Particle size of 181nm; Polydispersity index of 0.625; Zeta potential of -6.87mV and Entrapment efficiency of 84.85%. Further this formulation is evaluated by cytotoxicity study using MDA-MB-231 breast cancer cell line. The formulation AM-DOX liposome shows the least cell viability of 33.1% at 100μg/ml concentration compare with AM liposome and DOX liposome shows the cell viability of 53.76% and 47.8% at 100μg/ml concentration respectively, which shows that the formulation AM+DOX liposome shows the more significant percentage inhibition of MDA-MB-231 cell line. The AM-DOX conjugated liposomal drug delivery system significantly enhanced the delivery of Annona muricata leaf extract and Doxorubicin to the breast cancer cells and shows the least cell viability. FUTURISTIC STUDY: ➢ In-silico study shows only predicted values but scientific validation of these ligands requires further in-vitro and in-vivo animal studies. ➢ Further, these studies to be carried out in future by isolation of particular above mentioned compounds from Annona muricata and making the novel drug delivery formulation to overcome the P-gp efflux by inhibiting the multi-drug resistance proteins in the breast cancer to promote the therapeutic efficacy of the chemotherapeutic agent

Literature-based discovery of known and potential new mechanisms for relating the status of cholesterol to the progression of breast cancer

Breast cancer has been studied for a long period of time and from a variety of perspectives in order to understand its pathogeny. The pathogeny of breast cancer can be classified into two groups: hereditary and spontaneous. Although cancer in general is considered a genetic disease, spontaneous factors are responsible for most of the pathogeny of breast cancer. In other words, breast cancer is more likely to be caused and deteriorated by the dysfunction of a physical molecule than be caused by germline mutation directly. Interestingly, cholesterol, as one of those molecules, has been discovered to correlate with breast cancer risk. However, the mechanisms of how cholesterol helps breast cancer progression are not thoroughly understood. As a result, this study aims to study known and discover potential new mechanisms regarding to the correlation of cholesterol and breast cancer progression using literature review and literature-based discovery. The known mechanisms are further classified into four groups: cholesterol membrane content, transport of cholesterol, cholesterol metabolites, and other. The potential mechanisms, which are intended to provide potential new treatments, have been identified and checked for feasibility by an expert

The war against pain : the design, synthesis and testing of potential COX-2 selective inhibitors.

Ph. D. University of KwaZulu-Natal 2014.Much research has focused on the inhibition of the cyclooxygenase (COX) enzyme as this protein is responsible for the first step in the pain pathway in the conversion of arachidonic acid into prostaglandins and thromboxanes. The binding of curcumin and celecoxib, a known cyclooxygenase-2 (COX-2) selective inhibitor, was investigated computationally in order to identify important ligand-protein interactions which would need to be mimicked by a novel COX-2 selective compound. Initial investigations into the binding of curcumin identified the lesser diketone tautomer as having potential COX-2 selective activity. Two novel COX-2 selective compounds were designed using moieties common amongst known COX-2 selective compounds and moieties found in curcumin. Initial docking and binding scores showed that these compounds interacted in a similar manner with the protein as did celecoxib. Modifications to these initial compounds yielded two classes of compounds which explored the impact of the substitutions on the docking and binding scores, the poses and the ligand-protein interactions. All modifications made resulted in enhanced binding towards COX-2, and in a number of cases a reduction in the binding scores for COX-1. Thirty of the 166 compounds designed were selected for synthesis and biological screening as these compounds exemplified the range of changes observed in the full complement of compounds. Retrosynthesis yielded two potential synthetic pathways, and while the first path proved unsuccessful, the second route, which makes use of convenient reaction conditions, afforded the compounds in modest to good yields. Complete NMR spectroscopic analysis was carried out on all compounds, with Diffusion Ordered Spectroscopy used to determine the diffusion coefficients and hydrodynamic radii of two compounds and illustrated the dependence of these measurements on the properties of the medium. NMR Analysis of Molecular Flexibility In Solution (NAMFIS) analysis of one of the final compounds identified six conformers as existing in solution, based on the comparison of experimentally derived Nuclear Overhauser Enhancement (NOE) data with the results from a conformational analysis carried out in silico. Four of the six poses are responsible for >95% of the solution population, with one pose comprising almost 50%. All but one of the poses show Root Mean Squared Deviation (RMSD) values of less than 2 Å when compared to the predicted pose, indicating that any of these poses could bind into the protein. Initial inhibition screening results of the unsubstituted parent benzenesulfonate compound appeared to show three-fold selectivity of COX-2 over COX-1 at 100 nM. Testing of the substituted compounds revealed that these compounds are not COX-2 selective as desired, rather a number show promise as COX-1 selective compounds, with inhibition scores of over 40%, and several other compounds show potential as non-selective COX inhibitors. There is no obvious correlation between the inhibition results and either the Glide XP docking scores or the Prime binding scores, and as such, additional computational analysis as well as experimental testing is required to identify a correlation between the theoretical results and the experimental data, and illustrates that computational results cannot be the sole criterion on which selectivity is judged

Sewage sludge heavy metal analysis and agricultural prospects for Fiji

Insoluble residues produced in Waste Water Treatment Plants (WWTP) as by products are known as sewage sludge (SS). Land application of SS, particularly in agricultural lands, is becoming an alternative disposal method in Fiji. However, currently there is no legislative framework governing its use. SS together with its high nutrient and organic matter contents, constitutes some undesired pollutants such as heavy metals, which may limit its extensive use. The focus of this study therefore was to determine the total concentrations of Pb, Zn, Cd, Cu, Cr, Ni and Mn in the SS produced at the Kinoya WWTP (Fiji) and in the non-fertile soil amended with the SS at 20, 40, 60, 80% application rates and in the control (100% Soil). The bioavailable heavy metals were also determined as it depicts the true extent of metal contamination. The treatment mixtures were then used to cultivate cabbage plants in which the total heavy metal uptake was investigated. Total Zn (695.6 mg/kg) was present in the highest amounts in the 100% SS (control), followed by Pb (370.9 mg/kg), Mn (35.0 mg/kg), Cu (65.5 mg/kg), Cr (20.5 mg/kg) and finally Cd (13.5 mg/kg) and hence a similar trend was seen in all treatment mixtures. The potential mobility of sludgeborne heavy metals can be classified as Ni > Cu > Cd > Zn > Mn > Cr > Pb. Total metal uptake in plant leaves and stems showed only the bioavailable metals Cu, Cd, Zn and Mn, with maximum uptake occurring in the leaves. Ni, despite being highly mobile was not detected, due to minute concentrations in the SS treatments. Optimum growth occurred in the 20 and 40% SS treatments. However maximum Cu and Mn uptake occurred in the 40% SS treatment thereby making the 20% treatment the most feasible. Furthermore the total and bioavailable metal concentrations observed were within the safe and permitted limits of the EEC and USEPA legislations