Related Experiment Video
Updated: Aug 19, 2025
A Fluorescence-based Lymphocyte Assay Suitable for High-throughput Screening of Small Molecules
Published on: March 10, 2017
Pharmacophore-based virtual screening approaches to identify novel molecular candidates against EGFR through
F A Dain Md Opo1,2, Mohammed Moulay2,3,4,5, Ali Zari1,3
1Department of Biological Science, Faculty of Sciences, King Abdulaziz University, Jeddah, Saudi Arabia.
Abstract:
Alterations to the EGFR (epidermal growth factor receptor) gene, which primarily occur in the axon 18-21 position, have been linked to a variety of cancers, including ovarian, breast, colon, and lung cancer. The use of TK inhibitors (gefitinib, erlotinib, lapatinib, and afatinib) and monoclonal antibodies (cetuximab, panitumumab, and matuzumab) in the treatment of advanced-stage cancer is very common. These drugs are becoming less effective in EGFR targeted cancer treatment and developing resistance to cancer cell eradication, which sometimes necessitates stopping treatment due to the side effects. One in silico study has been conducted to identify EGFR antagonists using other compounds, databases without providing the toxicity profile, comparative analyses, or morphological cell death pattern. The goal of our study was to identify potential lead compounds, and we identified seven compounds based on the docking score and four compounds that were chosen for our study, utilizing toxicity analysis. Molecular docking, virtual screening, dynamic simulation, and in-vitro screening indicated that these compounds' effects were superior to those of already marketed medication (gefitinib). The four compounds obtained, ZINC96937394, ZINC14611940, ZINC103239230, and ZINC96933670, demonstrated improved binding affinity (-9.9 kcal/mol, -9.6 kcal/mol, -9.5 kcal/mol, and -9.2 kcal/mol, respectively), interaction stability, and a lower toxicity profile. In silico toxicity analysis showed that our compounds have a lower toxicity profile and a higher LD50 value. At the same time, a selected compound, i.e., ZINC103239230, was revealed to attach to a particular active site and bind more tightly to the protein, as well as show better in-vitro results when compared to our selected gefitinib medication. MTT assay, gene expression analysis (BAX, BCL-2, and β-catenin), apoptosis analysis, TEM, cell cycle assay, ELISA, and cell migration assays were conducted to perform the cell death analysis of lung cancer and breast cancer, compared to the marketed product. The MTT assay exhibited 80% cell death for 75 µM and 100µM; however, flow cytometry analysis with the IC50 value demonstrated that the selected compound induced higher apoptosis in MCF-7 (30.8%) than in A549.
Insights
New compounds targeting the epidermal growth factor receptor (EGFR) show promise in cancer treatment. These novel EGFR antagonists demonstrate superior efficacy and reduced toxicity compared to existing therapies, offering a potential advancement in cancer drug development.
Area of Science:
- Oncology
- Pharmacology
- Computational Chemistry
Background:
- Epidermal growth factor receptor (EGFR) gene alterations are implicated in various cancers, including lung and breast cancer.
- Current EGFR-targeted therapies like gefitinib face challenges with efficacy and drug resistance.
- Existing computational studies for EGFR antagonists lack comprehensive toxicity and comparative analyses.
Purpose of the Study:
- To identify novel EGFR antagonists with improved efficacy and reduced toxicity profiles.
- To computationally screen and validate potential lead compounds against EGFR.
- To compare the efficacy and safety of identified compounds with gefitinib.
Main Methods:
- Virtual screening and molecular docking to identify potential EGFR antagonists.
- In silico toxicity analysis and dynamic simulations to assess compound safety and stability.
- In vitro assays including MTT, apoptosis analysis, gene expression, and cell cycle analysis on lung and breast cancer cell lines.
Main Results:
- Four novel compounds (ZINC96937394, ZINC14611940, ZINC103239230, ZINC96933670) were identified with superior binding affinity and stability compared to gefitinib.
- In silico toxicity analysis revealed a lower toxicity profile and higher LD50 values for the identified compounds.
- In vitro studies demonstrated significant cell death and apoptosis induction, with ZINC103239230 showing enhanced efficacy over gefitinib in MCF-7 and A549 cell lines.
Conclusions:
- The identified compounds represent promising novel EGFR antagonists with enhanced efficacy and safety profiles.
- ZINC103239230 warrants further investigation as a potential therapeutic agent for EGFR-driven cancers.
- This study provides a validated computational and in vitro approach for discovering new cancer therapeutics.
