Rapid estimation of activation enthalpies for cytochrome‐P450‐mediated hydroxylations

Cytochrome P450 (CYP) enzymes play a critical role in detoxication and bioactivation of xenobiotics; thus, the ability to predict the biotransformation rates and regioselectivity of CYP enzymes toward substrates is an important goal in toxicology and pharmacology. Here, we present the use of the semiempirical quantum chemistry method SAM1 to rapidly estimate relative activation enthalpies (Δ H ‡ ) for the hydroxylation of aliphatic carbon centers of various substrates. The Δ H ‡ were determined via a reaction path calculation, in the reverse direction (RRP), using the iron‐hydroxo‐porphine intermediate and the substrate radical. The SAM1 Δ H ‡ were calculated via unrestricted Hartree‐Fock (UHF) and configuration interaction (CI) formalisms for both the doublet and quartet spin states. The SAM1 RRP Δ H ‡ , after subtracting a correction factor, were compared with density functional theory (DFT) B3LYP activation energies for two sets of substrates and showed R 2 ranging from 0.69 to 0.89, and mean absolute differences ranging from 1.2 ± 1.0 to 1.7 ± 1.5 kcal/mol. SAM1 UHF and CI RRP calculation times were, on average, more than 200 times faster than those for the corresponding forward reaction path DFT calculations. Certain key transition‐state (TS) geometry measurements, such as the forming O···H bond length, showed good correlation with the DFT values. These results suggest that the SAM1 RRP approach can be used to rapidly estimate the DFT activation energy and some key TS geometry measurements and can potentially be applied to estimate substrate hydroxylation rates and regioselectivity by CYP enzymes. © 2010 Wiley Periodicals, Inc. J Comput Chem, 2011