Drug Metabolism Phase II Transformations Acetyl Conjugation

Like methylation, acetylation results in a less hydrophilc metabolite (e.g.; an amine is converted into an amide). The physiological consequence is that the it may deactivate a drug or its Phase I metabolites (if they are active as well). Acetylation involves two steps: 1. Cofactor acetyl-CoA activates acetyltransferase 2. acetytransferase transfers the acetyl group to the substrate.

Drug Metabolism Phase II Transformations Acetyl Conjugation

cilastin is an inhibitor of dehydropeptidase, which hydrolyzes the antibacterial agent imipenem. Thus, cilastin prevents metabolism of imipenem when the two agents are coadministered.

Drug Metabolism Phase II Transformations Acetyl Conjugation

Sometimes acetyl conjugation occurs to detoxify substances (e.g. anilines) that could further produce other harmful metabolites!

Drug Metabolism Phase II Transformations Amino Acid Conjugation

Amino acid conjugation occurs for a variety of carboxylic acids and the result is the formation of an amide bond to an amino acid, usually glycine or glutamine. The reaction is catalyzed by amino acid acyl transferase enzymes. Glycine is the most common conjugating amino acid in mammals.

Drug Metabolism Phase II Transformations Amino Acid Conjugation

The mechanism of amino acid conjugation occurs in three steps: 1. 2. 3. activation of the carboxylic acid by ATP to generate the AMP ester conversion of the AMP ester to a coenyzme A thioester Acyl transferase-mediated coupling of the thioester with an amino acid.

Drug Metabolism Phase II Transformations Amino Acid Conjugation

Metabolism of brompheniramine (an antihistamine in Dimetapptm): Several Phase I metabolic processes are observed (N-oxidation, N-dealkylation, oxidative deamination, and aldehyde oxidation) to ultimately provide a carboxylic acid metabolite which is finally conjugated with glycine (Phase II).

Drug Metabolism Phase II Transformations Glucuronidation is one of the most common mammalian Phase II conjugation
pathways. There are three steps to form the conjugate: 1. coupling of D-glucose-1-phosphate to UTP to give UDP-glucose 2. oxidation of the primary alcohol yields the coenzyme UDP-glucuronic acid 3. conjugation with the substrate to yield the glucuronide

Drug Metabolism Phase II Transformations Glucuronidation example: metabolism of nortriptyline.

Note: the polar hydroxyl groups of the glucuronide impart great water solubility, which facilitates excretion!

Drug Metabolism Phase II Transformations Glucuronidation

There are 4 general classes of glucuronides that are produced: O-Glucuronide hydroxyl (alcohol, phenol) carboxylic acid N- Glucuronide amine amide sulfonamide : S- Glucuronide sulfhydryls (e.g. thiols) C- Glucuronide 1,3-dicarbonyls

R-SO2NH-R

R-SH

Drug Metabolism
Phase II Transformations O-Glucuronidation

Silverman, p. 331

Drug Metabolism
Phase II Transformations N-Glucuronidation

Silverman, p. 331

Drug Metabolism
Phase II Transformations S- and C-Glucuronidation

Silverman, p. 331

Drug Metabolism
Phase II Transformations

Sulfate Conjugation
Sulfate Conjugation is less prevalent than glucuronidation, presumably due to the lower availability of inorganic sulfate in mammals. The most common substrates that undergo sulfate conjugation are phenols.

Drug Metabolism
Phase II Transformations

Sulfate Conjugation
There 1. 2. 3. are three steps that occur with sulfate conjugation: activation of inorganic sulfate by ATP phosphorylation of the 3 OH to generate the sulfation cofactor. conjugation with the substrate to yield the sulfate.

Drug Metabolism
Phase II Transformations

Glutathione Conjugation
Glutathione (GSH) is a tripeptide and is found in nearly all mammalian tissues. Conjugation of this sort occurs in the cytoplasm of most cells, especially in the liver and kidney where the GSH concentration is 5-10 mM. Its apparent function is to scavenge potentially harmful electrophilic compounds; (either xenobiotics or their metabolites). This chemical basis for its function its reactive nucleophilic thiol group.

Drug Metabolism
Phase II Transformations Glutathione Conjugation
The conjugation is mediated by the enzyme glutathione transferase (GST) although with more reactive electrophiles the conjugation may occur nonenzymatically. Glutathione conjugation differs from the previous Phase II reactions since electrophiles (rather than nucleophiles) are subject to conjugation. Glutathione can to conjugate to many types of electrophilic species Well examine these two types: alkyl halide equivalents/epoxides (SN2 reactions!) activated aryl halides (SNAr reactions!)

Drug Metabolism
Phase II Transformations Glutathione Conjugation SN2-type reactions:

Drug Metabolism
Phase II Transformations Glutathione Conjugation
SNAr-type reactions: general mechanism

SNAr-type reactions: metabolism of azathiprine:

Drug Metabolism
Phase II Transformations Glutathione Conjugation
Once formed, GSH conjugates are rarely excreted directly but more often undergo a couple more steps of metabolism to be excreted as N-acetylcysteine conjugates. This pathway is considered by some to be Phase III metabolism.

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