Research Projects

Breast cancer remains a leading cause of cancer-related mortality worldwide, necessitating the development of novel, effective, and safer therapeutics. Conventional treatments often cause severe side effects, highlighting the need for alternative strategies. Both natural and synthetic pyrrole containing compounds are found to have varied biological activities. Some are particularly important for their anticancer activities. We have isolated and identified a novel small pyrrole containing compound namely (S)-1-(methoxycarbonyl)-2-methyl-2,5-dihydro-1H-pyrrole-2-carboxylic acid which is highly effective even at micromolar concentration against breast cancer cell but their inhibitory mechanism of action against breast cancer yet to be studied. Inspiring this molecule, we would synthesize pyrrole derivatives and perform preliminary screening for anti-breast cancer activity. We would study the cell killing potency of the selected compounds and the mechanisms they followed and the pathway would be elucidated by assaying different markers involved. Their effects on intracellular ROS generation, lipid peroxidation, cell cycle profile, DNA damage, anti-metastatic effect would be studied in breast cancer cell line and will compare with the positive (with conventional anti breast cancer drug) and negative controls (without treatment). Before going to the invitro study, the prediction of molecular mechanisms along with the determination of specific target site and the pathway involved in the processes, a systems-level perspective of drug–target–pathway interactions through network pharmacology would be follow which may accurately captures the biological complexity of cancer development. Thus, to know the molecular mechanism of the compound, network pharmacology can be established. Different publicly available servers, such as PharmMapper, Swiss Target Prediction, can be utilized to predict the drug target. Further, GeneCard, DisGeNET, and the Genetic Association Database will be used to identify disease targets. Thereafter, a Venn diagram will be created along with a PPI network to identify the most important target. The study of the network will be done on Cytoscape software. Functional enrichment analysis through KEGG and DAVID highlights key pathways involved. The compound(s) can be docked with target proteins using AutoDock Vina, and then binding affinities will be studied. GROMACS will be used to understand and to perform molecular dynamics simulations, to understand the modification of the compound(s) in physiological conditions. To understand the drug-like properties of the compound(s) ADMET study will be performed using SwissADME and pkCSM. Following computational validation, the promising compound(s) will be tested in an in vitro model. Initially apoptosis induction properties and IC₅₀ values will be calculated. Based on RNA-sequencing we would determine the differential gene expression profiling in treated breast cancer cell lines, and identify different modulation of major cancer-associated pathways such as protein kinase, DNA replication, DNA repairing, cell cycle pathways etc. Also to validate the results obtained from network studies, the expression of target specific genes will be evaluated through RT-PCR, and the primers will be designed based on the in-silico experiment. Western blot experiment will be performed to understand the target protein expression after treating with the drug-like molecule(s). These comprehensive studies including sequence-based genome wide differential mRNA expression of treated cells compare with untreated control, network pharmacology and target specific gene expression using RT-PCR may discover novel targets. Thus, discovery of such promising drug like molecules with effective anticancer activities may overcome the drawbacks of conventional drugs if successfully translated into medicine for cancer patients.