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ADME Predictions and Safety Profiles of Indole Derivatives as Potential Kinase Inhibitors

INTRODUCTION The physical, biological, and toxicological properties of a drug candidate are directly linked to its structure. The knowledge and awareness of the role of unwanted pharmacokinetic properties of drugs has been noticed and problematic toxicophores are issued as major problems for drug development [1]. For this reason, study on the ADMET properties of drug candidates has become essential in the drug development process to eliminate inappropriate pharmacokinetic profile of compounds [2]. In addition, metabolites with physicochemical and pharmacological properties that differ substantially from those of the parent drug consequently have important implications for both drug safety and efficacy. In fact, ~30% of the attrition in drug discovery and development is still due to safety concerns. To reduce the risk of costly clinical-stage due to metabolic characteristics of drug candidates, there is a need for efficient and reliable ways to predict drug metabolism as well as identify the pitfalls in current approaches to estimating and predicting adverse effects (AEs) and toxicity in humans [3]. Modelling and simulation techniques have become a standard tool for the evaluation of Pharmacokinetic (PK) properties, supporting the extrapolation and prediction of drug disposition and treatment effects in humans. Src Family Tyrosine Kinase Inhibitors (SFKIs) have been designed as anti-cancer agents in past several years and several small molecules were approved by FDA for clinical studies [4]. It was shown that various substituted 2oxindole derivatives have the ability to inhibit several SFKs [5]. In this study, in silico study was applied to predict the cytochrome P450 (CYP) metabolism and toxicity parameter of substituted 2-oxindole derivatives that were previously synthesized and tested on tyrosine kinase activity [6-8]. MATERIALS AND METHODS 2-Oxindole derivates were evaluated in silico by using ADMET PredictorTM (Simulations Plus Inc.). ADMET Predictor was used with its Metabolism module, i.e., CYP substrate/non substrate classification models for CYP isoforms 1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1 and 3A4, their corresponding site of metabolism models (SOM) as well as kinetic parameter models (Km, Vmax and Clint) and CYP inhibition models for 1A2, 2C9, 2C19, 2D6, and 3A4. Metabolites were generated using SMIRKS strings (a reaction transform language) to specify the transformations predicted by the SOM models. The CYP kinetic parameters were predicted from ANNE regression models. RESULTS AND DISCUSSION The results of this study revealed an extensive CYP metabolism of 2-oxindole derivatives mediated mainly by three CYP isoforms (2C9, 2D6 and 3A4) with percentage of metabolized molecules as it follows: CYP3A4_Substr > CYP2C9_Substr > CYP2D6_Substr (81.8% > 31.8% > 27.3%, respectively) and CYP2C9_Inh > CYP3A4_Inh > CYP2D6 (86.6% > 29.5% > 4.5%, respectively). The inhibition of testosterone metabolism by CYP3A4 was predicted for 32 of 44 molecules (CYP3A4_Inh 72.7%) which could have an implication of potential drug-drug interactions of these compounds. The most represented metabolic reactions mediated by CYPs were: aromatic hydroxylation, oxidative Ndealkylation of di-substituted urea- and thiourea-2-oxindoles to their corresponding mono-substituted metabolites. For all thiourea derivatives the reaction of desulphuration was also predicted by which -NH-C(=S)-NH- moiety was converted into urea moiety what implies a potential pro-drug use of thiourea-2oxindoles (Fig. 1). Figure 1. Metbolic pathways of thourea-2oxindole derivatives. All thiouerea 2-oxindoles were metabolized to corresponding urea metabolites what implies their use as pro-drugs, however, for all thiourea-2-oxindoles the highest CYP_Risk scores and TOX_Risks, including mutagenicity and/or carcinogenicity were predicted (Fig. 2). Figure 2. Predicted Toxicity risk of investigated 2-oxindoles CONCLUSION Extensive CYP metabolism by CYP2C9, 2D6 and 3A4 were predicted for Src indole derivatives. Investigated 2-oxindoles with good in vitro activity and predicted IC50 values, physico-chemical and toxicity parameters were found comparable to reference compound imatinib. REFERENCES 1. Hisaka A., et al. Prediction of pharmacokinetic drug-drug interaction caused by changes in cytochrome P450 activity using in vitro formation. Pharmacol. Ther. 2010. 125: 230-248. 2. Jones BC. et al. Combination of docking, molecular dynamics and quantum mechanical calculations for metabolism prediction of 3, 4-methylenedioxybenzoyl2-thienylhydrazone. Prog Med. Chem. 2009. 47: 239–263. 3. Andrade HC. et al. In silico Prediction of Drug Metabolism by P450. Curr. Drug Metab. 2014.15: 514-525. 4. Roskoski R. Src protein-tyrosine kinase structure, mechanism, and small molecule inhibitors. Pharmacol. Res. 2015. 94: 9-25. 5. Zhang S., Yu D. Targeting Src family kinases in anti-cancer therapies: turning promise into triumph. Trends Pharmacol. Sci. 2012. 33: 122-128. 6. Kilic-Kurt Z. et al. Synthesis, biological, and computational evaluation of novel 1, 3, 5-substituted indolin-2-one derivatives as inhibitors of src tyrosine kinase. Arch. Pharm. Chem. Life Sci. 2015. 348: 1-15. 7. Kilic-Kurt Z. et al. Synthesis, biological and computational evaluation of novel oxindole derivatives as inhibitors of src family kinases. LDDD. 2013. 10: 713-718. 8. Kilic-Kurt Z. et al. Synthesis and pp60csrc tyrosine kinase inhibitory activity of novel indole-3-imine derivatives and their amine congeners substituted at N1 and C5. Arch. Pharm. 2009. 342: 333-343.

ADME, SFKIs, tyrosine kinase inhibitors, 2-oxindoles, CYP metabolism

Rad je rezultat znanstvene suradnje Farmaceutsko-biokemijekog fakulteta Sveučilišta u Zagrebu i Farmaceutskog Fakulteta Biruni sveučilišta u Istanbulu (Turska)