Novel 3,4-diaminothieno[2,3-b]thiophene-2,5-dicarbohydrazide-based scaffolds as EGFRWT, EGFRT790M, and tubulin polymerization inhibitors with anti-proliferative activity

Using 3,4-diaminothieno[2,3-b]thiophene-2,5-dicarbohydrazide 5, new scaffolds were created that inhibit EGFR^WT, EGFR^T790M protein kinases, and tubulin polymerization. Target compounds were evaluated for anti-proliferative efficacy toward A431, H1975, and A549 cells via the MTT technique. The results indicated that compounds 7a, 7b, and 9 exhibited the most significant anti-proliferative activity, exhibiting IC₅₀ values ranging from 4.64 to 9.45 μM, surpassing the potency of erlotinib, gefitinib, and osimertinib. Compound 7a inhibited EGFR^T790M and EGFR^WT kinases via IC₅₀ values of 24.6 and 28.1 nM, respectively. The outcomes exceeded those of osimertinib and gefitinib, which had IC₅₀ values of 58.1, 8.1, 17.8, and 373.5 nM. Derivative 7a showed significant anti-proliferative effects on HCT116 and T47D cells, via IC₅₀ values of 3.12 and 4.12 μM, respectively. Analog 7a selectively suppressed HEK293 cells and hampered tubulin polymerization through IC₅₀ values of 5.1 μM, comparable with CA-4 (IC₅₀ = 4.3 μM). The synthetic analogs (7a, 7b, and 9) DOCK at the EGFR^WT, EGFR^T790M, and tubulin active binding sites, matching their in vitro data. Analogs 7a and 7b, like osimertinib and neratinib, can create an irreversible covalent binding with Cys797 in the EGFR^T790M ATP cleft, potentially treating mutations. MD simulations by the iMODs server evaluated protein-7a complex stability. In silico ADME analysis of prospective EGFR inhibitors 7a and 9 utilizing the egg-boiled technique showed favorable lipophilicity, GIT absorption, and BBB permeability. Thus, our proposed compounds 7a and 9 showed promising anti-proliferative properties, targeting EGFR^WT and EGFR^T790M kinases and inhibiting tubulin.

Anti-cancer agents; Dual EGFR(WT); EGFR(T790M) inhibitors; Hydrazide; Tubulin inhibitors.