Xeno
Xeno
Some xenobiotics cause toxicity by disrupting normal cell functions: Bind and damage proteins (structural, enzymes) Bind and damage DNA (mutations)
Mutagenesis - UV light
Carcinogenesis - benzene, asbestos Teratogenesis - thalidomide
Metabolism of xenobiotics Chemical transformation of xenobiotics Occurs in mostly in liver (enzymatic processes) Convertion into more hydrophil. subst. - excretion urine
Introduction
Purpose
Converts lipophilic to hydrophilic compounds Facilitates excretion
Consequences
Detoxification Metabolic activation
Biotransformation
Potentially toxic xenobiotic
Relatively harmless
Detoxification
Metabolic activation
Inactive metabolite
Reactive intermediate
Xenobiotic
Phase I - Activation Reactive intermediate Phase II - Conjugation Conjugate
(non-polar)
Excretion
Phase I
introduction of functional group
hydrophilicity increases slightly may inactivate or activate original compound major player is CYP or mixed function oxygenase (MFO) system in conjunction with NAD(P)H location of reactions is smooth endoplasmic reticulum
Phase II
conjugation with endogenous molecules
(GSH, glycine, cystein, glucuronic acid)
hydrophilicity increases substantially neutralization of active metabolic intermediates facilitation of elimination location of reactions is cytoplasm
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Phase I reactions
Oxidation
Hydroxylation (addition of -OH group) N- and O- Dealkylation (removal of -CH side chains) Deamination (removal of -NH side chains) Epoxidation (formation of epoxides) Oxygen addition (sulfoxidation, N-oxidation) Hydrogen removal
O C C
epoxide
Reduction
Hydrogen addition (unsaturated bonds to saturated) Donor molecules include GSH, FAD, NAD(P)H Oxygen removal
Hydrolysis
Splitting of amide and ester bonds
Drug Metabolism
Metabolic enzymes
1. Microsomal:
1. 2. CYP450 monooxygenases Flavin monooxygenase
2. Non-microsomal
1. 2. 3.
1. 2. 3.
3. Both
CYP genes have multiple alleles (2D6 has 53, and 2E1 has 13)
N Fe N
N N
Heme
Different forms of heme are found in hemoglobins and proteins known as cytochromes.
The iron in a heme can bind oxygen and transfer electrons.
Cytochrome P450 Cytochrome P450s are integral membrane proteins found in the endoplasmic reticulum-membrane network in cells
NADP+
CYP
Drug
NADPH CO
CYP-Fe+2 Drug
CO hu
H2O 2H+
CYP Nomenclature
Families - CYP plus arabic numeral (>40% homology of amino acid sequence, eg. CYP1) Subfamily - 40-55% homology of amino acid sequence; eg. CYP1A Subfamily - additional arabic numeral when more than 1 member has been identified; eg. CYP1A2 Italics indicate gene (CYP1A2); regular font for enzyme
CYP Biotransformations
Chemically diverse small molecules are converted, generally to more polar compounds Reactions include:
Aliphatic hydroxylation, aromatic hydroxylation Dealkylation (N-,O-, S-) N-oxidation, S-oxidation Deamination Dehalogenation
O C HN CH3
OH Acetaminophen
At normal doses, sulfate or a sugar is attached to OH and the drug is easily removed by the kidney.
At high doses, this pathway cannot keep up and a liver P450 converts acetaminophen to a toxic metabolite which causes liver damage.
Other Conjugation Reactions: O-Methylation, SMethylation, Amino Acid Conjugation (glycine, taurine, glutathione) Many conjugation enzymes exhibit polymorphism
Require
molecular
oxygen,
NADPH,
flavin
adenosine
dinucleotide (FAD) FMOs are heat labile and metal-free, unlike CYPs Factors affecting FMOs (diet, drugs, sex) not as highly studied as CYPs
HO
These are released into the blood where they bind to receptors on the surface of cells.
To ensure that the action of amine neurotransmitters is short, they are destroyed by the enzyme monoamine oxidase.
HO NH3 tyramine
Foods containing high levels of tyramine: smoked, aged, or pickled meat or fish; sauerkraut aged cheeses (e.g., swiss, cheddar, blue, boursault, camembert, emmenthaler, stilton); yeast extracts fava beans beef or chicken liver aged sausages (e.g., bologna, pepperoni, salami, summer sausage) game meats (e.g., venison, rabbit) red wines (e.g., chianti, sherry).
Monoamine oxidase
HO HO NH3
O2
HO HO O + NH4
R' N N
O NH
Monoamine oxidase (MAO) inhibitors are used as antidepressants and to treat eating disorders. They keep serotonin levels up. Serotonin controls mood. MAO inhibitors can cause overdoses: No longer break down tyramine from diet. High levels of tryptophan in diet can result in serotonin levels becoming too high. Will prevent metabolism of drugs that are amines. As a result MAO inhibitors can result in serious side effects when used with other drugs. MAO inhibitors have been replaced with drugs which prevent the binding of serotonin to cells, such as Prozac. These do not affect the metabolism of other amines, so that there are far fewer side effects.
Phase I: Hydrolysis
Carboxyesterases & peptidases
hydrolysis of esters eg: valacyclovir, midodrine hydrolysis of peptide bonds e.g.: insulin (peptide)
Epoxide hydrolase
H2O added to epoxides eg: carbamazepine
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Phase I: Reductions
Azo reduction
N=N to 2 -NH2 groups eg: prontosil to sulfanilamide
Nitro reduction
N=O to one -NH2 group eg: 2,6-dinitrotoluene activation
Reductions
Carbonyl reduction
Alcohol dehydrogenase (ADH)
Chloral hydrate is reduced to trichlorothanol
Disulfide reduction
First step in disulfiram metabolism
Sulfoxide reduction
NSAID prodrug Sulindac converted to active sulfide moiety
Reductions
Quinone reduction
Cytosolic flavoprotein NAD(P)H quinone oxidoreductase
two-electron reduction, no oxidative stress high in tumor cells; activates diaziquone to more potent form
Flavoprotein P450-reductase
one-electron reduction, produces superoxide ions metabolic activation of paraquat, doxorubicin
Reductions
Dehalogenation
Reductive (H replaces X)
Enhances CCl4 toxicity by forming free radicals
Phase I: Oxidation-Reduction
Alcohol dehydrogenase
Alcohols to aldehydes Genetic polymorphism; Asians metabolize alcohol rapidly Inhibited by ranitidine, cimetidine, aspirin
Aldehyde dehydrogenase
Aldehydes to carboxylic acids Inhibited by disulfiram
Phase I: Monooxygenases
Monoamine oxidase
Primaquine, haloperidol, tryptophan are substrates Activates 1-methyl-4-phenyl-1,2,5,6tetrahydropyridine (MPTP) to neurotoxic toxic metabolite in nerve tissue, resulting in Parkinsonian-like symptoms
Monooxygenases
Peroxidases couple oxidation to reduction of H2O2 & lipid hydroperoxidase
Prostaglandin H synthetase (prostaglandin metabolism)
Causes nephrotoxicity by activating aflatoxin B1, acetaminophen to DNA-binding compounds
Monooxygenases
Flavin-containing mono-oxygenases
Generally results in detoxification Microsomal enzymes Substrates: nicotine, cimetidine, chlopromazine, imipramine Repressed rather than induced by phenobarbital, 3-methylcholanthrene
Phase II reactions
Glycoside conjugation - glucuronidation Sulfate - sulfation Glutathione (GSH) Methylation Acylation
Acetylation Amino acid conjugation Deacetylation
Phosphate conjugation
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Phase 2: Conjugation
Glucose
UDP-Glucuronate HO OH
Carboxylic acids
1,3-Dicarbonyls
RXH CO2H X O OH OH OH R Kidney Urive excretion CO2H X O OH OH OH R
Poor reabsorb.
RXH
CO2H X O OH OH OH
Sulfation
Sulfation is a high affinity, low capacity pathway
Glucuronidation is low affinity, high capacity
Aromatic -NH2 or NHOH conjugated with COOH of serine, proline, requiring ATP activation
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