Reversible inhibition modalities,

If the inhibitor is found to bind rapidly (linear progress curves) and dissociate rapidly (rapid recovery of activity upon dilution) from its target enzyme, then one can proceed to analyze its inhibition modality and affinity by classical methods. The modes of reversible inhibition of enzymes were described in Chapter 3. In the next section of this chapter we will describe convenient methods for determining reversible inhibition modality of lead compounds and lead analogues during compound optimization (i.e., SAR) studies. 

DETERMINING REVERSIBLE INHIBITION MODALITY AND DISSOCIATION CONSTANT... 

Determining Reversible Inhibition Modality and Dissociation Constant 129... 

By experiments performed as discussed here, the reversible inhibition modality of each lead compound, representing a distinct pharmacophore or chemical structural class, can be defined and the dissociation constant (A) or aK<) can be determined from the data fitting to Equations (3.1) through (3.6). As lead analogues are produced within a structural series, one can generally assume that the inhibiton modality will be the same as that of the founder molecule (i.e., the lead) of that structural series. This assumption simplifies the determination of dissociation constants for other series molecules, as described below. However, this assumption must be... 

In this chapter we described the thermodynamics of enzyme-inhibitor interactions and defined three potential modes of reversible binding of inhibitors to enzyme molecules. Competitive inhibitors bind to the free enzyme form in direct competition with substrate molecules. Noncompetitive inhibitors bind to both the free enzyme and to the ES complex or subsequent enzyme forms that are populated during catalysis. Uncompetitive inhibitors bind exclusively to the ES complex or to subsequent enzyme forms. We saw that one can distinguish among these inhibition modes by their effects on the apparent values of the steady state kinetic parameters Umax, Km, and VmdX/KM. We further saw that for bisubstrate reactions, the inhibition modality depends on the reaction mechanism used by the enzyme. Finally, we described how one may use the dissociation constant for inhibition (Kh o.K or both) to best evaluate the relative affinity of different inhibitors for ones target enzyme, and thus drive compound optimization through medicinal chemistry efforts. 

---