IN SILICO SCREENING AND IN VITRO TESTING OF POTENTIAL MULTITARGET TYROSINE KINASE INHIBITORS OF EGFR, HER2, AND VEGFR2

• Main Author: Sangande, Frangky
• Format: Dissertations
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/56547
• Institution: Institut Teknologi Bandung

Cancer is still a major public health issue worldwide. In the cancer treatment, cytotoxic agents are commonly used class of drugs. However, these agents lack selectivity, causing side effects in their use. Therefore, targeted therapies have been developed that work more selectively by targeting specific molecules that play a role in cancer development such as EGFR (epidermal growth factor receptor), HER2 (human EGFR-related receptor 2), and VEGFR2 (vascular endothelial growth factor receptor 2). They represent validated targets for cancer therapy. EGFR and HER2 are members of the tyrosine kinase receptor that belong to the same family. Both are synergistic in their signal transduction processes to regulate cell proliferation, differentiation, migration, and apoptosis. In pathological conditions, coexpression of EGFR and HER2 has been reported to be assosiated with poor prognosis and leads to EGFR inhibitors resistant. On the other hand, VEGFR2 plays a role in supporting the growth of cancer cells through the process of angiogenesis. This receptor has also been reported to participate in cases of resistance to EGFR inhibitors. Therefore, simultaneous inhibition of EGFR, HER2, VEGFR2 is considered to increase the efficacy and overcome the resistance problem. Based on this, the present study was designed to identify potential compounds as multitargeted EGFR, HER2, and VEGFR2 inhibitors by in silico and in vitro method. The study was initiated by conducting in silico screening of ZINC database using validated pharmacophore modeling and molecular docking. From this screening, a hit compound, called ZINC21942954, was predicted to have inhibitory activity against the three targets. Using the scaffold of ZINC21942954, a structural similarity search was carried out against the MCULE database to obtain compounds with a Tanimoto coefficient ?0.7 and re-screened them using molecular docking. The final resluts obtained three additional hit compounds, called ZINC35560729, ZINC35372090, and ZINC35560766 which predicted their inhibitory activity was better than ZINC21942954. The four compounds were further analyzed for their interaction stability against the three targets using molecular dynamics simulations and also calculated their free energy using MMPBSA method. The study was continued by testing their in vitro inhibitory activity at a single concentration of 1 µM. The test results showed that ZINC21942954 had multitarget EGFR and VEGFR2 inhibitory activity. Its inhibitory activity against VEGFR2 was moderate while against EGFR was weak. The other hit compounds showed multitarget EGFR and HER2 inhibitory activity. The order of inhibitory potency for each compound against both targets was ZINC35560729 > ZINC35372090 > ZINC35560766. ZINC35560729 had EGFR inhibitory activity at a moderate level, while against HER2 was strong. Meanwhile, two other compounds showed weak inhibitory activity, except against HER2, ZINC35372090 showed moderate inhibitory activity. Based on the above results, ZINC35560729 can be considered as a potential candidate for EGFR and HER2 dual inhibitor. This compound had inhibitory activity against EGFR and HER2 with IC50 values of 5.02 µM and 0.83 µM, respectively. In addition, this compound was also selective in targeting EGFR and HER2. Thus, ZINC35560729 can be used as a lead compound for further development.