From analyses of human P450 substrates and their physicochemical properties, it is apparent that baseline lipophilicity relationships exist for over 70 substrates of eight drug-metabolizing P450 enzymes from families CYP1, CYP2, and CYP3. Equations of the general form shown below result in all cases investigated thus far: deltaG(bind) = adeltaG(part) + b where a is the slope of the line which can be termed the hydrophobicity factor of the enzyme active site, possibly being related to the extent of hydrophobic amino acid residues lining the heme pocket; b is the intercept on the y axis and can be regarded as the sum of nonhydrophobic interactions between enzyme and substrate; deltaG(bind) is the free energy change for substrate binding to P450, based on the relationship deltaG(bind) = RTlnKm where Km is the Michaelis constant, and deltaG(part) is the free energy change for partitioning between n-octanol and water based on the relationship deltaG(part) = -RTlnP where P is the n-octanol/water partition coefficient. These findings facilitate the analysis of P450 enzyme-substrate binding interactions and provide information about the likely hydrophobic character of human P450 active site regions. This shows that there are common interactions for certain numbers of substrates in each case composed of hydrogen bonding and pi-pi stacking, the extent of which varies from one P450 enzyme to another.