Inhibition slowly reversible/irreversible inhibitors

Studies in recent years have revealed a number of remarkable drug interactions with irreversible or mechanism-based inhibitors of CYP3A, many of which can be attributed to inhibition of sequential intestinal and hepatic first-pass metabolism. Mechanism-based inhibition involves the metabolism of an inhibitor to a reactive metabolite, which either forms a slowly reversible metabolic-intermediate (MI) complex with the heme moiety or inactivates the enzyme irreversibly via covalent binding to the enzyme catalyzing the last step in the bioactivation sequence. As a result, mechanism-based inhibition is both... 

An important feature of enzymes is that their active sites can often be occupied by, or react with, molecules other than the substrate, leading to inhibition of enzyme activity. Several inhibition mechanisms are known, but it is necessary only to distinguish between irreversible and reversible inhibition. Irreversible inhibition arises when the inhibitor molecule I dissociates very slowly or not at all from the enzyme active site. The best-known examples occur when I reacts covalently with a critical residue in the active site. Inhibition of cholinesterase enzymes by the reaction of organo-phosphorus compounds with a serine residue is a case in point. This type of inhibition is said to be noncompetitive—enzyme activity cannot be restored by addition of excess substrate. So although addition of I reduces V,n x, Km is unaffected. The double-reciprocal plot in such cases has the same. v-axis intercept as the plot for the uninhibited enzyme, but greater slope. 

It should be recalled here that the alcoholic hydroxyl of serine does not possess a dissociation constant within the pH range, accessible to enzymic reactions. Therefore, this amino acid cannot influence the pH-activity curve. On the other hand, it is well known that DFP inhibition is initially reversible and becomes only slowly irreversible. This has been demonstrated for true ChE from electric eel by Nachmansohn and associates (46) and for plasma ChE by Mackworth and Webb (47). Similarly, a stepwise reaction with inhibitors, containing the diethyl phosphoryl moiety, has been made probable by Hobbiger (34)- Therefore, it appears possible that phosphates are first attacked by the imidazol moiety of the esteratic site, in conformity with the catalytic influence of free imidazol on phosphate hydrolysis (48). This step is followed by transfer to serine. The final product is a trialkyl phosphate XV, which is not split by imidazol (scheme F). 

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