5-Lipoxygenase (5-LOX) plays a key role in the pathway of leukotriene biosynthesis. To predict the inhibitory activity of flavonoid inhibitors against 5-LOX from Spatholobus suberectus Dunn, Autodock 4.2 and comparative molecular field analysis (CoMFA) were employed. For the positive inhibitors (n=7), the value of the coefficient of determination ( R 2 ) between the binding free energy, calculated using Autodock 4.2, and the experimental pIC 50 is 0.838. In the training set (n=21) of inhibitors against 5-LOX, the R 2 of non-cross-validated partial least squares analysis between the actual and predicted pIC 50 values, using the no-validation with the optimum number of components set to 6, is 0.997 (p=0.000). For the model generated by CoMFA, the contribution of electrostatic and steric factors are 0.522 and 0.478, respectively. Among the flavonoids of S. suberectus , liquiritigenin, catechin, butin, 3′,4′,7-trihydroxyflavone, plathymenin, and gallocatechin are the more potent inhibitors of 5-LOX based on the calculated binding free energy and the predicted pIC 50 value.