Design, synthesis, and evaluation of novel quinazoline derivatives as potent EGFR inhibitors: In silico and in vitro approaches.

Searching for novel epidermal growth factor receptor (EGFR) inhibitors, 1-substituted 3,5-diphenyl pyrazolines 4a-i, 5a-i, 6a, and 6b bearing the terminal piperidine or morpholine moieties commonly observed in clinically approved EGFR inhibitors were synthesized as novel anti-cancer agents acting via EGFR inhibition. A series of 3,5-diphenyl pyrazolines was synthesized and screened for in vitro anti-cancer activity against 60 NCI cell lines. Pyrazolines 5d and 6a revealed broad-spectrum cytotoxic activities and potent EGFR inhibition with IC50 values of 2.30 µM and 1.47 µM, respectively, in comparison to Vandetanib (IC50 = 0.5 µM) and Gefitinib (IC50 = 0.04 µM). Interestingly, compound 6a demonstrated a promising cytotoxic activity against the leukemia cell line (HL-60) and safety toward the normal cell line HSF. Additionally, compound 6a up-regulated proapoptotic markers and down-regulated Bcl-2 as an antiapoptotic marker in HL-60 cells. Docking simulations explained the EGFR inhibitory actions of 5d and 6a compared to Gefitinib. According to predictive models of oral bioavailability and drug-likeness, pyrazolines 5d and 6a are expected to be bioavailable and drug-like compounds. Pyrazolines 5d and 6a are novel EGFR inhibitors with a broad-spectrum anti-cancer activity, and 6a has off-target antileukemic effect.

6,7-Dimorpholinoalkoxy quinazoline derivatives as potent EGFR inhibitors with enhanced antiproliferative activities against tumor cells

Cancer, particularly non-small cell lung cancer (NSCLC), remains a major global health challenge, necessitating the development of novel and more effective therapeutic agents. Epidermal growth factor receptor (EGFR) inhibitors, such as erlotinib, have shown clinical efficacy in inhibiting EGFR-mediated tumor growth. However, the emergence of resistance mechanisms and concerns regarding bioavailability highlight the need for more potent and selective EGFR inhibitors. Molecular docking has emerged as a valuable tool in the early stages of drug discovery, providing insights into ligand-receptor interactions and predicting binding affinities. This review evaluates the accuracy of molecular docking by comparing in silico binding predictions with in vitro cytotoxicity data. Erlotinib, a well-established EGFR inhibitor, serves as a reference for assessing the performance of docking simulations. We further explore the structure-activity relationship (SAR) of novel 1,3-diazetidin-2-one derivatives as potential EGFR inhibitors, highlighting key pharmacophoric features that contribute to enhanced binding and specificity. Despite the utility of docking studies, limitations such as neglecting pharmacokinetic factors and metabolic stability are discussed. The review concludes by proposing strategies to integrate docking with in vitro and in vivo testing to improve the accuracy of predictive models and optimize lead compounds for clinical development

Cancer is a major health problem to human beings around the world. Many quinazoline derivatives were reported to have potent cytotoxic activity. Our aim in this work is the discovery of potent epidermal growth factor receptor (EGFR) inhibitors with anti-breast cancer activity containing 4-substituted quinazoline pharmacophore. Novel series of 4-substituted 6,8-dibromo-2-(4-chlorophenyl)-quinazoline derivatives have been designed and synthesized. New derivatives were tested against MCF-7 (human breast carcinoma cell line) and screened for their inhibition activity against epidermal growth factor receptor tyrosine kinase (EGFR-TK). Most of the tested compounds show potent antiproliferative activity and EGFR-TK inhibitory activity. Compounds VIIIc and VIIIb exerted powerful cytotoxic activity (IC50 3.1 and 6.3 μM) with potent inhibitory percent (91.1 and 88.4%) against EGFR-TK. Compounds IX, VIIa, X, VIIb, VIc, V, IV, VIa and VIb showed promising cytotoxic effects with IC50 range (12-79 μM) with good activity against EGFR-TK with the inhibitory percent (85.4-60.8%). On the other hand, compounds VIIc, VIIIa exerted low cytotoxic effects as revealed from their IC50 value (124 and 144 μM) with low activity against EGFR-TK with inhibitory percent 30.6 and 29.1% respectively.

Epidermal growth factor receptor (EGFR) has been recognized as one of the attractive targets for anticancer drug development. Herein, a set of anilino-1,4-naphthoquinone derivatives (3–18) was synthesized and investigated for their anticancer and EGFR inhibitory potentials. Among all tested compounds, three derivatives (3, 8, and 10) were selected for studying EGFR inhibitory activity (in vitro and in silico) due to their most potent cytotoxic activities against six tested cancer cell lines (i.e., HuCCA-1, HepG2, A549, MOLT-3, MDA-MB-231, and T47D; IC50 values = 1.75–27.91 μM), high selectivity index (>20), and good predicted drug-like properties. The experimental results showed that these three promising compounds are potent EGFR inhibitors with nanomolar IC50 values (3.96–18.64 nM). Interestingly, the most potent compound 3 bearing 4-methyl substituent on the phenyl ring displayed 4-fold higher potency than the known EGFR inhibitor, erlotinib. Molecular docking, molecular dynamics simulation, and MM/GBSA-based free energy calculation revealed that van der Waals force played a major role in the accommodations of compound 3 within the ATP-binding pocket of EGFR. Additionally, the 4-CH3 moiety of the compound was noted to be a key chemical feature contributing to the highly potent EGFR inhibitory activity via its formations of alkyl interactions with A743, K745, M766, and L788 residues as well as additional interactions with M766 and T790.

Based on the analysis of epidermal growth factor receptor (EGFR) complexes with gefitinib with molecular docking, the scaffold-hopping strategy, combination of the active substructures, and structural optimization of EGFR inhibitors, a novel series of benzo[4,5]imidazo[2,1-b]thiazole derivatives was designed, synthesized, and evaluated for antitumor activity in human cancer cell lines and cellular toxicity against human normal cell lines using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay and EGFR inhibitory activities in vitro. Some target compounds such as 2-(benzo[4,5]imidazo[2,1-b]thiazol-3-yl)-N-(2-hydroxyphenyl)acetamide (D04) and 2-(benzo[4,5]imidazo[2,1-b]thiazol-3-yl)-N-(naphthalen-1-yl)acetamide (D08) have shown significant antitumor activity against the EGFR high-expressed human cell line HeLa. All the target compounds showed hardly any antitumor activity against the EGFR low-expressed human cell line HepG2, and nearly no cellular toxicity against the human normal cell lines HL7702 and human umbilical vein endothelial cell lines (HUVEC). The inhibitory activities against EGFR kinase in vitro of the three target compounds were greatly consistent with the anti-proliferative activities. The preliminary structure–activity relationships of the target compounds were summarized. Conclusively, the novel benzo[4,5]imidazo[2,1-b]thiazole derivatives as novel potential EGFR inhibitors may be used as the potential lead compounds for the development of antitumor agents.

Aberrant activation of the epidermal growth factor receptor (EGFR) is closely associated with malignant progression of tumors. EGFR inhibitors have been used successfully in clinic in the treatment of solid tumors. In the present study, we revealed that BB，a new synthetic quinonazoline derivative，was a potent EGFR inhibitor. BB selectively inhibited EGFR with a IC50 value of 50±37 nM, at least 32-fold more potent than suppressed all other 10 tested receptor tyrosine kinases including the same family member ErbB2 (IC50=5.6±3.2 μM). BB effectively abrogated autophosphorylation of the EGF-stimulated EGFR and phosphorylation of its key downstream signaling molecules ERK and AKT in A549 cells. BB was shown to suppress EGF-stimulated proliferation of A549 cells with an apparently lower IC50 value (0.33±0.07 μM) than that (2.7±0.4μM) for the serum-stimulated cells. BB also inhibited the EGF-independent proliferation of a panel of tumor cells. In addition, BB exhibited anti-angiogenesis activity, as evidenced by antagonizing EGF-induced HMEC-1 migration in vitro, blocking HMEC-1 tube formation, and inhibiting microvessel sprouting from rat aortic rings. Most importantly, BB prominently inhibited in vivo tumorigenesis of NIH3T3 cells specifically driven by the activation-mutated EGFR genes. As reported, normal NIH3T3 cells lack tumorigenicity in nude mice. NIH3T3 cells transfected with the EGFR gene with activating mutation (A750P or L858R) produced rapidly growing xenografts in nude mice. BB, when given orally at 100 mg/kg consecutively for 2 weeks, prominently inhibited the growth of the xenografts and reduced the number of microvessels. Taken together, the data indicate that BB is a new selective EGFR inhibitor with potent antitumor activity, revealing its potential as a promising anticancer candidate.

Novel 4-anilinoquinazoline derivatives featuring an 1-adamantyl moiety as potent EGFR inhibitors with enhanced activity against NSCLC cell lines

Design, synthesis and biological evaluation of structurally new 4-indolyl quinazoline derivatives as highly potent, selective and orally bioavailable EGFR inhibitors

Progress in identifying and understanding the molecular and cellular causes of cancer has led to the discovery of anomalies that characterize cancer cells and that represent targets for the development of cancer therapeutics. One such target is the epidermal growth factor receptor (EGFR), a transmembrane protein that is frequently dysregulated in cancer cells and associated with the development, progression and aggressiveness of a number of malignancies. Inhibition of EGFR signaling has thus been identified as an attractive strategy in control of tumor proliferation, and over a decade of intense activity in the field has culminated in the discoveries and subsequent approvals of gefitinib and erlotinib for the treatment of non-small cell lung cancer. However, the drug's resistance to gefitinib and erlotinib has been clinically observed. Therefore, intensive efforts have been made in the discovery of novel potent and selective EGFR inhibitors. This review will focus on the developments of small molecule EGFR inhibitors based on the quinazoline core scaffolds in recent 5 years. Diverse EGFR inhibitors are classified as 4-anilinoquinazolines and 4-nonanilininoquinazolines, their biological data are described, and the structure-activity relationships (SARs) are discussed.

Designing strategies, structural activity relationship and biological activity of recently developed nitrogen containing heterocyclic compounds as epidermal growth factor receptor tyrosinase inhibitors

Background: The active mutation of epidermal growth factor receptor (EGFR), L858R or deletion in exon19, are present in patients with NSCLC and responsive to EGFR tyrosine kinase inhibitors such as gefitinib and erlotinib. Over time, most tumors develop resistance to these EGFR inhibitors by the acquiring of drug-resistant mutations including gatekeeper T790M mutation (more than 50% cases). Severe skin rash and diarrhea caused by wild-type EGFR (wtEGFR) inhibition limit EGFR TKI treatment. Therefore targeting mutant EGFR (mtEGFR) but not wtEGFR represents an attractive therapeutic strategy. In this report, we identify a small molecule EGFR inhibitor, TAS-121, that targets mtEGFR but not wtEGFR and its biological characterization in vitro. Methods: In vitro enzyme inhibition activity of TAS-121 for EGFR and its mutants was determined by using purified EGFRs and peptide substrates. Cellular phosphorylation of EGFR was assayed by immunoblot analysis. For growth inhibition assay, cells were treated with TAS-121 for 3 days, and living cells were determined by using CellTiter GloTM which measures cellular ATP. Apoptosis assay was conducted by using Caspase-GloTM and Cell Counting Kit. Results: TAS-121 was found to be a more potent mutant selective EGFR-TKI based on bioavailability characteristics compared to our proto-type compound, TAS-2913. TAS-121 inhibits kinase activity of EGFR mutants harboring T790M in a sub-nano molar range and much more potently than that of wtEGFR. TAS-121 inhibits auto-phosphorylation of EGFR and intercellular signaling pathways in NCI-H1975 (EGFR T790M / L858R) and HCC827 (EGFR del E746-A750). In a cell proliferation assay, TAS-121 also suppresses growth of NCI-H1975 and HCC827 more potently than that of a normal cell which grows in an EGF-dependent manner. Therefore, the wtEGFR / mtEGFR inhibition ratio of TAS-121 appears to be higher than those of other EGFR TKIs. Furthermore, TAS-121 induces apoptosis in NCI-H1975 but erlotinib could not. Conclusion: These results suggest that TAS-121, a potent mutant-selective EGFR inhibitor, may represent a new therapeutic strategy in the treatment of NSCLC resistant to currently available EGFR/TKIs. Further pre-clinical evaluations and development are ongoing. Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C111. Citation Format: Kimihiro Ito, Kazutaka Miyadera, Yoshimi Aoyagi, Masanori Kato, Kazuhiko Yonekura, Yoshikazu Iwasawa, Teruhiro Utsugi. In vitro characterization of TAS-121, a novel, highly potent, and mutant-specific EGFR-TKI. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C111.

The epidermal growth factor receptor (EGFR) presents a crucial target for combatting cancer mortality. This study employs a suite of computational techniques, including 3D-QSAR, ligand-based virtual screening, molecular docking, fingerprinting analysis, ADME, and DFT-based analyses (MESP, HOMO, LUMO), supplemented by molecular dynamics simulations and MMGB/PBSA free energy calculations, to explore the binding dynamics of quinazoline derivatives with EGFR. With strong q2 and r2 values from CoMFA and CoMSIA models, our 3D- QSAR models reliably predict EGFR inhibitors' efficacy. Utilizing a potent model compound as a reference, an E-pharmacophore model was developed to sift through the eMolecules database, identifying 19 virtual screening hits based on ShapeTanimoto, ColourTanimoto, and TanimotoCombo scores. These hits, assessed via 3D- QSAR, showed pIC50 predictions consistent with experimental data. Our analyses elucidate key features essential for EGFR inhibition, reinforced by ADME studies that reveal favorable pharmacokinetic profiles for most compounds. Among the primary phytochemicals examined, potential EGFR inhibitors were identified. Detailed MD simulation analyses on three select ligands-1Q1, 2Q17, and VS1-demonstrated their stability and consistent interaction over 200 ns, with MM/GBSA values corroborating their docking scores and highlighting 1Q1 and VS1's superior EGFR1 affinity. These results position VS1 as an especially promising lead in EGFR1 inhibitor development, contributing valuable insights towards crafting novel, effective EGFR1 inhibitors.

Rational design of novel potential EGFR inhibitors by 3D-QSAR, molecular docking, molecular dynamics simulation, and pharmacokinetics studies

Design, preparation and biological evaluation of new Rociletinib-inspired analogs as irreversible EGFR inhibitors to treat non-small-cell-lung cancer