synthesis of which endogenous substrates share enzymes associated with metabolism of xenobiotics
steroid hormones
Vitamin D cogeners
bile acids
Vitamin D cogeners
bile acids
usual mechanism of phase I reactions
convert drug to more polar metabolite by introducing or unmasking functional group
typical functional groups used in phase I rxns
OH
NH2
SH
NH2
SH
hallmarks of phase II rxns
combines glucuronic acid, sulfurinc acid, acetic acid, or amino acid with the polar functional group added in phase I
principal organ of drug metabolism
liver
other organs involved in drug metabolism
GI
skin
kidneys
lungs
brain
skin
kidneys
lungs
brain
location of subcellular enzymes used in drug transformations
ER
mitochondria
cytosol
lysosomes
nuclear envelope
plasma membrane
mitochondria
cytosol
lysosomes
nuclear envelope
plasma membrane
MFO/monooxygenases
responsible for oxidative drug metabolism
require reducing agent (NADPH) and O2
require reducing agent (NADPH) and O2
typical MFO rxn in general
one O2 consumed per substrate molecule, 1 O in product and 1 in H2O
NADPH-cytochrome P450 reductase is made of
FMN and FAD
cytochrome P450
hemoprotein
terminal oxidase
inc with repeat exposure to drug
terminal oxidase
inc with repeat exposure to drug
what is the rate limiting step in hepatic drug oxidations
P450 heme reduction
the reductase is less abundant than the hemoprotein
the reductase is less abundant than the hemoprotein
what do microsomal drug oxidations require
P450, P450 reductase, NADPH, O2
general steps of microsomal drug oxidations
1. oxidized (Fe3+) P450 + drug substrate= binary complex
2. NADPH donates electron to P450 reductase (reduces the P450-drug complex)
3. NADPH adds another e- to reductase, reducing O and making an activated O P450-substrate complex
4. transfers the activated O to the drug
2. NADPH donates electron to P450 reductase (reduces the P450-drug complex)
3. NADPH adds another e- to reductase, reducing O and making an activated O P450-substrate complex
4. transfers the activated O to the drug
what is the common feature of drugs oxidized by the p450 system
high lipid solubility
most important forms of liver P450 enzymes
CYP1A2
CYP2A6
CYP2B6
CYP2C9
CYP2D6
CYP2E1
CYP3A4
CYP2A6
CYP2B6
CYP2C9
CYP2D6
CYP2E1
CYP3A4
which P450 enzyme is responsible for over 50% of the Rx drugs metabolized in liver
CYP3A4
how is P450 expression induced
inc rate of synthesis or dec rate of degradation
sources of other organic compounds that induce CYP1A
tobacco smoke, charred meat, organic pyrolysis products
chemicals that induce CYP3A
dexamethasone
rifampin
mifepristone
phenobarbital
atorvastatin
hyperforin (St. John's Wort)
rifampin
mifepristone
phenobarbital
atorvastatin
hyperforin (St. John's Wort)
basic mechanism of induced P450 synthesis
AhR (cytoplasmic receptor) binds with polycyclic aromatic hydrocarbons-> into nucleus-> dimer with Arnt-> activation of reg elements of CYP1A genes
CAR
constitutive adrostane receptor
used for phenobarbital class of inducers for CYP2B6, CYP2C9, CYP3A4
used for phenobarbital class of inducers for CYP2B6, CYP2C9, CYP3A4
PPARalpha
nuclear receptor highly expressed in liver and kidneys
lipid lowering drugs as ligands
eg- fenofibrate, gemfibrozil
mediates induction of CYP4A
lipid lowering drugs as ligands
eg- fenofibrate, gemfibrozil
mediates induction of CYP4A
RXR
retinoid X receptor
forms heterodimer with PXR, CAR, and PPARalpha upon their binding their ligand
forms heterodimer with PXR, CAR, and PPARalpha upon their binding their ligand
CYP4A is responsible for
fatty acid metabolism
eg arachidonic acid
eg arachidonic acid
PXR
pregnane X receptor
steroid-retinoid-thyroid receptor family
mediates CYP3A induction
steroid-retinoid-thyroid receptor family
mediates CYP3A induction
substrate stabilization
induction of P450s by dec degradation
how does imidizole inhibit P450
bind tightly to P450 heme iron and reduce substrate metabolism thru competitive inhibition
drug ex: cimetidine, ketoconazole
drug ex: cimetidine, ketoconazole
how do macrolide antibiotics inhibit P450
metabolized by CYP3A, metabolites complex the P450 heme iron and render it catalytically inactive
suicide inhibitors
inactivators that attack the heme or protein moiety
drug conjugates
drugs coupled with endogenous substance (phase II rxn)
polar molecules that are readily excreted and often inactive
polar molecules that are readily excreted and often inactive
where are transferases located
microsome or cytosol
which transferases are the most dominant
UGTs
Uridine 5'diphosphate glucuronosyl transferases
Uridine 5'diphosphate glucuronosyl transferases
mechanisms of SULTs
sulfotransferases
sulfation of substrates using PAPS (3'phosphoadenosine 5'phosphosulfate- sulfur donor)
sulfation of substrates using PAPS (3'phosphoadenosine 5'phosphosulfate- sulfur donor)
GSH
glutathione
GST
glutathione transferases
metab drugs, xenobiotics, leukotrienes, prostaglandins
metab drugs, xenobiotics, leukotrienes, prostaglandins
NATs
cytosolic N-acetyltransferases
use acetyl CoA as cofactor for metabolizing chemicals with aromatic amine or hydrazine
use acetyl CoA as cofactor for metabolizing chemicals with aromatic amine or hydrazine
SAMe
S-adenosyl-L-methionine
mediates O, N, and S methylation of drugs/xenobiotics using methyltransferases
mediates O, N, and S methylation of drugs/xenobiotics using methyltransferases
EHs
epoxide hydrolases
hydrolyze endobiotic, drug, and xenobiotic epoxides from P450 oxidations
hydrolyze endobiotic, drug, and xenobiotic epoxides from P450 oxidations
what plays a critical role in the regulation of drug conjugations
nutrition because endogenous substrates originate in teh diet
normal mechanism of acetaminophen metabolism
glucuronidation and sulfation (95%) and P450 dependent GSH conjugation (5%)
when does acetaminophen become toxic
when levels saturate the P450 pathway, no hepatic GSH available for conjugation, toxic metabolite accumulates, damages liver
why is GSH not effective antidote for acetaminophen induced hepatotoxicity
doesn't readily cross cell membranes
2 effective antidotes to acetaminophen overdosage
cysteamine
N-acetylcysteine (safer)
N-acetylcysteine (safer)
what factors influence individual variations in drug distribution, metabolism, and elimination
genetic factors, age, sex, liver size, liver function, circadian rhythm, temperature, nutrition, environment (exposure to inducers/inhibitors of drug metabolism)
individual differences in metabolic rate depend on
the nature of the drug
slow acetylator phenotype
50% blacks/whites in US, more freq in Europeans at high altitudes
higher incidence of drug-induced autoimmune disorders and bicyclic aromatic amine-induced bladder cancer
higher incidence of drug-induced autoimmune disorders and bicyclic aromatic amine-induced bladder cancer
genetic polymorphisms in UGT are associated with
hyperbirubinemic diseases and impaired drug conjugation/elimination
genetic polymorphisms in GST associated with
significant adverse effects and toxicities of drugs dependent on GSH conjugation pathway
debrisoquin-sparteine oxidation polymorphism
3-10% whites, auto recessive
CYP2D6 oxidations of debrisoquin, etc are impaired
faulty expression of the P450 protein
CYP2D6 oxidations of debrisoquin, etc are impaired
faulty expression of the P450 protein
ultrarapid metabolism
CYP2D6 polymorphism with up to 13 repeat genes in tandem
common in Ethiopians and Saudi Arabians
common in Ethiopians and Saudi Arabians
CYP2C19 polymorphism
stereoselective aromatic 4-hydroxylation of mephenytoin
auto recessive
3-5% caucasians, 18-23% Japanese
poor metabolizers completely lack enzyme activity
auto recessive
3-5% caucasians, 18-23% Japanese
poor metabolizers completely lack enzyme activity
CYP2C9 polymorphism
CYP2C9*2- impaired functional rxns with P450 reductase
CYP2C9*3- lowered affinity for many substrates
CYP2C9*3- lowered affinity for many substrates
CYP2C9*3 polymorphism would affect metabolism of which drugs
warfarin, henytoin, losartan, tolbutamide, some NSAIDs
significance of low CYP2A6 activity
smokers with this gene consume less and have lower incidence of lung cancer
Ziegler's enzyme
falvin monooxygenase
polymorphisms with low activity result in fish-odor syndrome
polymorphisms with low activity result in fish-odor syndrome
charbroiled foods and cruciferous vegetables induce which enzyme
CYP1A
grapefruit juice inhibits which enzyme
CYP3A
examples of enzyme-inducing drugs
sedative-hypnotics
antipsychotics
anticonvulsants
rifampin (anti-TB)
insecticides
antipsychotics
anticonvulsants
rifampin (anti-TB)
insecticides
75% of biotransformation from these 2 enzymes
CYP3A4
CYP2D6
CYP2D6