Unadkat Vishal, Rohit Shishir, Parikh Paranjay, Patel Kaushal, Sanna Vinod, Singh Sanjay
Kashiv Biosciences Pvt Ltd, Ahmedabad, 382210, Gujarat, India.
Division of Biological & Life Sciences (Formerly Institute of Life Sciences), School of Arts & Sciences, Ahmedabad University, Ahmedabad, 380009, Gujarat, India.
Onco Targets Ther. 2022 May 2;15:479-495. doi: 10.2147/OTT.S357765. eCollection 2022.
In this work, we have identified heterocyclic derivatives with 1,2,4 oxadiazole scaffold mimicking the functions of tyrosine kinase inhibitors. Fourteen molecules that displayed the best fit were picked from the library of compounds and studied under in-silico and in-vitro conditions. Four compounds were selected for further cytotoxicity and ADME (Absorption, Distribution, Metabolism, Elimination) profiling showing IC (from 8-13 µM) values against EGFR positive cancer cell line (MCF7).
A molecular dynamics simulation study was performed to understand the correlation of non-covalent binding energies with biological activity. The drug-like properties of the selected four compounds (7a, 7b, 7e, and 7m) were evaluated by in-vitro ADME studies. Compounds , and were the active compounds in the molecular dynamics simulations study. Further, EGFR binding activity was confirmed with EGFR and EGFR kinase assay using a luminescence-based method.
These compounds (, and ) showed activity against EGFR and mutant EGFR, exhibiting IC values of <10 and <50 micromolar, respectively. These compounds also possess moderate aqueous solubility in 40-70 µg/mL at pH 7.4 and 30-100 µg/mL at pH 4.0. Further, , and showed balanced lipophilicity with Log D values ranging from 1-3. They demonstrated a good correlation in Caco-2 permeability with Apparent permeability (Papp) 1 to 5 × 10 cm/s in comparison with , which was found to be highly lipophilic (Log D >5) and showed high permeability (Papp 17 × 10 cm/s). Lastly, all these compounds were moderately stable in liver microsomes at alkaline pH with a half-life of 30-60 min, while at a highly acidic pH (2.0), the compounds were stable up to 15-20 min.
Overall, in-vitro ADME results of these molecules showed good drug-like properties, which are well correlated with the in-silico ADME data, making them ideal for developing an oral drug delivery formulation.
在本研究中，我们鉴定了具有1,2,4-恶二唑骨架的杂环衍生物，其模拟酪氨酸激酶抑制剂的功能。从化合物库中挑选出14个拟合度最佳的分子，并在计算机模拟和体外条件下进行研究。选择了4种化合物进行进一步的细胞毒性和ADME（吸收、分布、代谢、排泄）分析，结果显示其对表皮生长因子受体（EGFR）阳性癌细胞系（MCF7）的半数抑制浓度（IC）值为8 - 13µM。
进行了分子动力学模拟研究，以了解非共价结合能与生物活性之间的相关性。通过体外ADME研究评估了所选4种化合物（7a、7b、7e和7m）的类药性质。化合物7a、7b和7e是分子动力学模拟研究中的活性化合物。此外，使用基于发光的方法通过EGFR和EGFR激酶测定法确认了EGFR结合活性。
这些化合物（7a、7b和7e）对EGFR和突变型EGFR均有活性，其IC值分别<10和<50微摩尔。这些化合物在pH 7.4时还具有40 - 70µg/mL的中等水溶性，在pH 4.0时为30 - 100µg/mL。此外，7a、7b和7e显示出平衡的亲脂性，Log D值范围为1 - 3。与Log D>5且具有高渗透性（表观渗透率Papp为17×10 cm/s）的7m相比，它们在Caco - 2通透性方面具有良好的相关性，表观渗透率（Papp）为1至5×10 cm/s。最后，所有这些化合物在碱性pH的肝微粒体中具有中等稳定性，半衰期为30 - 60分钟，而在高酸性pH（2.0）下，化合物在15 - 20分钟内保持稳定。
总体而言，这些分子的体外ADME结果显示出良好的类药性质，与计算机模拟ADME数据具有良好的相关性，使其成为开发口服给药制剂的理想选择。
