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5. IMPLICATIONS FOR DRUG METABOLISM
1.
Termination of drug action
2.
Activation of prodrug
3.
Bioactivation and toxication
4.
Carcinogenesis
5.
Tetratogenesis
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6. Termination of Drug Action
atropine
propranolol
(active)
tropic acid and tropine
→
hydroxypropranolol
(active)
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7. Termination of Drug Action
Conversion of drug to active metabolite to active
metabolite to inactive metabolite
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9. Inactive Terfenadine is Converted to its Active
Metabolite Fexofenadine
activation of prodrug
terfenadine
fexofenadine
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10. Some Xenobiotics Are Metabolized to Carcinogenic Agents
carcinogenesis
• 3,4 Benzopyrene
• Aflatoxin
• N-Acetylaminoflluorene
Metabolites of these agents interact with DNA
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11. Small Amounts of Acetaminophen is Converted to the
Reactive Metabolite N-Acetylbenzoquinoneimine
bioactivation
Bioactivation of acetaminophen; under certain conditions, the electrophile Nacetylbenzoquinoneimine reacts with tissue macromolecules, causing liver necrosis.
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12. Thalidomide is a Teratogen
teratogensis
– THALIDOMIDE: Fetal malformations in
humans, monkeys, and rats occur due to
metabolism of the parent compound to a
teratogen. This occurs very early in gestation.
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14. Factors Affecting Drug Metabolism
•
•
•
•
•
•
•
•
•
•
Age
Diet
Genetic Variation
State of Health
Gender
Degree of Protein Binding
Species Variation
Substrate Competition
Enzyme Induction
Route of Drug Administration
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15. Factors Affecting Drug Metabolism
• Route of drug administration
– Oral versus systemic administration
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16. Many Drugs Undergo First Pass Metabolism
Upon Oral Administration
• Oral administration
• Drug travels from gut to portal vein to liver
• Vigorous metabolism occurs in the liver. Little drug
gets to the systemic circulation
• The wall of the small intestine also contributes to first
pass metabolism
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17. ORGAN SITES OF DRUG METABOLISM
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18. Organ Sites of Drug Metabolism
•
•
•
•
•
•
•
Liver
Small intestine
Kidney
Skin
Lungs
Plasma
All organs of the body
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22. 80
Velocity Of Metabolism Of A Drug
70
Velocity
(ng/g tissue/min)
60
50
40
30
20
10
0
0
10
20
30
40
[Drug] mM
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50
60
70
D:summer1Kmx1.pzm
23. Velocity Of Metabolism Of A Drug
80
70
Velocity
(ng/g tissue/min)
60
zero order metabolism
50
40
30
20
10
0
0
first order metabolism
5 10 15 20 25 30 35 40 45 50 55 60
[Drug] mM
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Kmx2.pzm
24. First Order Metabolism
A drug may be given in doses that produce blood
concentrations less than the Km of the enyzme for the drug.
v = Vmax [C]
Km + [C]
When
then
Km >>> [C],
v = Vmax [C] ,
Km
and
v α [C]
Metabolism of the drug is a first order process. A constant
fraction of the remaining drug is metabolized per unit time.
Most drugs are given at concentrations smaller than the Km
of the enzymes of their metabolism.
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25. Velocity Of Metabolism Of A Drug
80
70
Velocity
(ng/g tissue/min)
60
zero order metabolism
50
40
30
20
10
0
0
first order metabolism
5 10 15 20 25 30 35 40 45 50 55 60
[Drug] mM
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Kmx2.pzm
26. Zero Order Metabolism
A drug may be given in doses that produce blood concentrations
greater than the Km of the enyzme for the drug.
v = Vmax [C]
K m + [C]
When [C] >>> Km,
then
v = Vmax [C] ,
[C]
and
v = Vmax
Metabolism of the drug is a zero order process. A constant
amount of the remaining drug is metabolized per unit time.
Phenytoin undergoes zero order metabolism at the doses
given.
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27. Velocity Of Metabolism Of A Drug
80
70
Velocity
(ng/g tissue/min)
60
zero order metabolism
50
40
30
20
10
0
0
first order metabolism
5 10 15 20 25 30 35 40 45 50 55 60
[Drug] mM
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Kmx2.pzm
28. Velocity
(ng/g tissue/min )
Velocity Of Metabolism Of Three Drugs
By The Same Enzyme
70
60
50
Drug A
Drug B
Drug C
40
30
20
10
0
0
10
20
30
40
50
[Drug] mM
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60
70
80
90
32. Phase II Metabolism
A molecule endogenous to the body donates a portion
of itself to the foreign molecule
D+ENDOX
DX+ENDO
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33. Patterns of Drug Metabolism
• Parent molecule → Phase 1 metabolism
• Phase 1 metabolite → Phase 2 metabolism
• Parent molecule → Phase 2 metabolism
• Phase 2 metabolite → Phase 1 metabolism
Some drugs are not metabolized, for example, gallamine
and decamethonium. Atracurium undergoes spontaneous
hydrolysis.
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56. CYP3A4
• CYP3A4 is responsible for metabolism of 60%
of all drugs
• It comprises approximately 28% of hepatic
cytochrome P450
• Metabolizes terfenadine
• Ingestion of grapefruit juice reduces expression
of this enzyme
• Inhibited by some regularly used drugs
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57. Some Drugs That Inhibit CYP3A4
• Macrolide antibiotics
– Erythromycin
– Clarithromycin
– Other such agents
• Antifungal agents
– Ketoconazole
– Itraconazole
– Other such agents
• HIV protease inhibitors
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58. CYP3A4
• Ketoconazole and terfenadine can produce a
drug interaction with fatal consequences.
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64. CYP3A4 And P-Glycoprotein
• P-Glycoprotein and CYP3A4 control oral bioavailability
of many drugs
• P-Glycoprotein and CYP3A4 share many substrates
and inhibitors
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65. CYP2D6 is an Enzyme with Polymorphisms
• Approximately 70 nucleotide polymorphisms are
known
• Four phenotype subpopulations of metabolizers*
– Poor metabolizers (PM)
– Intermediate metabolizers (IM)
– Extensive metabolizers (EM)
– Ultrarapid metabolizers (UM)
• Variations according to racial background
• More than 65 commonly used drugs are
substrates
• Codeine is a well known substrate
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* The Pharmacological Basis of Therapeutics
66. Codeine is a Substrate of CYP2D6
-CH3
(methyl morphine)
Consider the variation in codeine’s metabolism among
PM, IM, EM, UM individuals
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67. CYP2C9
• Metabolizes some 16 commonly used drugs
• Warfarin and phenytoin are among the substrates
• Two allelic variants are known: metabolizes substrates
5% to 12% of the wild type enzyme
– Warfarin clearance is greatly reduced in individuals
possessing the allelic variants
• Dose adjustments are required for drugs in individuals
who have the mutant enzymes
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68. CYP2C19
• S-mephenytoin is a substrate
– (4-hydroxylation at the phenyl ring)
• As much as eight allelic variants identified
– All are nonfunctional proteins
• Poor metabolizers of S-mephenytoin lack 4-hydroxylase
activity, but N-demethylation to nirvanol is an alternative
but slow metabolic pathway
– Dose adjustments must be made for poor
metabolizers of S-mephenytoin and for other drugs
that are substrates for this enzyme
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69. CYP1A1
• Polycyclic hydrocarbons are among its
substrates
• Inducers include
– Polycyclic hydrocarbons such as 3,4,-benzopyrene,
3-methylcholanthrene, etc.
– Charcoal broiled foods (polycyclic hydrocarbons)
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72. Nonmicrosomal Oxidations
Alcohol dehydrogenation is conducted by the enzyme
alcohol dehydrogenase (cytosolic)
Aldehyde dehydrogenation is conducted by the enzyme
aldehyde dehydrogenase (cytosol and mitochondria)
Xanthine oxidation is conducted by the cytosolic enzyme
xanthine oxidase.
Diamine oxidase (cytosolic) oxidizes histamine and
diamines such as cadaverine and putrescine.
Monoamine oxidation is conducted by mitochondrial
monoamine oxidase (norepinephrine, epinephrine,
dopamine and serotonin are endogenous substrates.
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74. Some Popular Substrates of Monoamine Oxidase
•
•
•
•
•
Serotonin
Epinephrine
Norepinephrine
Dopamine
Tyramine (found in certain foods)
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76. Alcohol Dehydrogenase
• A soluble enzyme, found almost exclusively in the
parenchymal cells of the liver
• Converts ethanol to acetaldehyde
• Converts methanol to formaldehyde
• Converts ethylene glycol to its respective aldehyde
metabolites
• Is inhibited by pyrazole
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97. PHASE 2 METABOLISM
A molecule endogenous to the body donates a portion
of itself to the foreign molecule
D+ENDOX
DX+ENDO
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100. Uridine-5’-α-D-glucuronic Acid
The microsomal enzyme glucuronyl transferase conducts the
donation of glucuronic acid from the endogenously synthesized
UDPGA to various substrates to form glucuronide conjugates.
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Examples of such substrates are morphine and acetaminophen.
101. UDP-α-D-Glucuronsyltransferase
•
•
•
•
•
Is also called glucuronyl transferase
A microsomal enzyme
Substrates are called aglycones
Conducts phase 2 metabolic reactions
Products are called glucuronides
• Glucuronides formed
–
RN-G; RO-G; RCOO-G; RS-G; RC-G
• Bilirubin is an endogenous substrate
• Induced by phenobarbital
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107. Induction Of UDP-α-D-Glucuronyl Transferase
• Induced by phenobarbital
• Induced by 3-methylcholanthrene
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108. Glucuronidation in the Cat
• The cat can glucuronidate bilirubin but cannot
glucuronidate phenolic compounds such as phenol
and napthol
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110. Sulfate Conjugation
• Conducted by the soluble enzyme sulfotransferase
• Endogenous donor molecule to conjugation is
3’-phosphoadenosine-5’-phosphosulfate (PAPS)
• Conjugates are ethereal in character
• Noninducible
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111. 3’-Phosphoadenosine-5’-phosphosulfate (PAPS)
The cytosolic enzyme sulfotransferase conducts the donation of
sulfate from the endogenously synthesized PAPS to various
substrates to form sulfate conjugates. An example of such substrate
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is acetaminophen.
112. Sulfate Conjugation of p-Hydroxyacetanilid
PAP: 3’-phosphoadenosine- 5’-phosphate
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114. Species Differences in Sulfate Conjugation
• Some species are deficient in the sulfate conjugation
pathway
– Pig
– Opposum
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116. N-Acetyltransferase
• A soluble enzyme
• Isoniazid is a substrate
• Genetic variation occurs
– Some individuals are fast acetylators
– Some individuals are slow acetylators
• Acetyl coenzyme A is the endogenous donor
molecule
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117. Acetyl CoA
Various acetylases, for examples, choline acetylase and N-acetyl
transferase, all soluble enzymes, conduct the transfer of the acetyl
group of acetyl CoA to various substrates. For example, N-acetylation
of isoniazid. Genetic polyporphism occurs with N-acetyltransferase.
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119. N-Acetyltransferase
• The dog cannot acetylate aromatic amino
compounds because it lacks the appropriate
isoenzyme of NAT
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123. S-Adenosylmethionine
Cytosolic enzymes such as catechol-O-methyl transferase (COMT) and
phenylethanolamine-N-methyl transferase (PNMT) conducts the
donation of the methyl group from the endogenously synthesized SAM
to various substrates to form methylated conjugates. Norepinephrine is
N-methylated by PNMT to form epinephrine. Norepinephrine,
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epinephrine, dopamine, and L-DOPA are O-methylated by COMT.
124. Methyltransferases
• A family of soluble enzymes that conducts
–
–
–
N-methylation; N-CH3
O-methylation; O-CH3
S-methylation; S-CH3
• S-adenosylmethionine (SAM)is the endogenous donor
molecule. It is demethylated to S-adenosylhomocysteine
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128. S-Methylation of 6-Mercaptopurine
TPMT - thiopurinemethyltransferase; some individuals are
deficient in this enzyme that is critically important for the
metabolism of this agent
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129. METABOLISM OF MERCAPTOPURINE (1)
6-Mercaptopurine
TMPT
6-Methylmercaptopurine
• TMPT -Thiomethylpurinetransferase
– Conducts S-methylation of the substrate
– Found in RBC’s
– Isoforms exist
• active enzyme
• inactive enzyme
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132. Multiple Metabolic Pathways Exist
for Aspirin’s Metabolism
Hydolysis of aspirin produces salicyclic acid, as
seen in the next slide
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133. Salicyluric Acid is the Glycine Conjugate of Aspirin
Salicyluric acid, the glycine conjugate of salicyclic acid, is the main
metabolite of aspirin. Approximately 76% of aspirin is metabolized
through amino acid conjugation.
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134. Acetyl Salicylic Acid (Aspirin) Metabolism
• Salicylic acid the hydrolytic product of acetyl salicylic
acid. Salicylic acid is further metabolized
• Salicyl uric acid is the glycine conjugate and the main
metabolite of aspirin. About 75% of aspirin is
metabolized by this pathway
• Other metabolites of aspirin
– the acyl glucuronide conjugate of salicylic acid (salicylic acid
glucuronide)
– the phenol glucuronide conjugate of salicylic acid (salicyl phenol
glucuronide)
– the ring hydroxylated product of salicylic acid (gentisic acid)
– the ring hydroxylated product of the glycine conjugate (gentisuric
acid
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138. DRUG INTERACTION WITH GLUTATHIONE
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mercapturate
metabolite of drug
139. MERCAPTURIC ACID FORMATION
• Conjugation of substrate to glutathione by the
enzyme glutathione transferase
• Hydrolytic removal of glutamic acid by glutamyl
transpeptidase
• Hydrolytic removal of glycine by cysteinyl glycinase
• Acetylation of the cysteinyl substrate by
N-acetyltransferase to form the N-acetylated cysteinyl
conjugate of substrate; substrate referred to as a
“mercapturate”
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142. ACETAMINOPHEN AND ITS PHASE II METABOLITES
The sulfate and glucuronide conjugates of acetaminophen are the major
metabolites. High doses of acetaminophen can exhaust the metabolic pathways
that produce these conjugates, allowing more of the parent drug to undergo the
phase I metabolic pathway which is involved in bioactivation and toxication.
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144. ACETAMINOPHEN AND ITS PHASE I METABOLITES- pt2
The minor metabolite (4% of acetaminophen), N-hydroxyacetaminophen,
is always produced by microsomal cytochrome P450. It rearranges to
the electrophile N-acetylbenzoquinoneimine, which in turn reacts with
the sulfhydryl group of glutathione. Acetaminophen mercapturic acid is
the final metabolite. If tissue glutathione stores are depleted as a result
of fasting, intake of excessive doses of acetaminophen or through
induction of CYP2E1 as a result of chronic intake of ethanol, the
quinone interacts with nucleophilic sites of cellular macromolecules,
such as proteins. Liver necrosis is the result. Regular intake of
acetaminophen during fasting or chronic ethanol intake should be
avoided. N-acetylcysteine is the antidote for acetaminophen poisoning.
It reacts with the electrophile. A small amount of acetaminophen is
reported to undergo deacetylation to the phase 1 metabolite paminophenol.
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148. Further Metabolism of N-HydroxyAAF Produces Cancer
N-HydroxyAAF undergoes phase II metabolism to the
ultimate carcingogen. The glucuronide pathway is also
involved in carcinogenesis
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154. CORRELATION BETWEEN SLEEPING TIME AND PLASMA
T1/2 IN CHRONIC PENTOBARBITAL PRETREATED RABBITS
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155. Factors Affecting Drug Metabolism
• Enzyme Induction - increased enzyme protein levels
in the cell
– Phenobarbital type induction by many drugs
– Polycyclic hydrocarbon type induction by
polycyclic hydrocarbons such as 3,4-benzopyrene
and 3-methylcholanthrene
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159. FACTORS AFFECTING DRUG METABOLISM
• Diet
– Charcoal broiled foods (contain polycyclic
hydrocarbons that increase certain enzyme protein in
cells)
– Grapefruit juice (the active component is the
furancoumarin 6,7-dihydroxybergamottin which
inhibits a certain a group of microsomal enzymes)
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163. FACTORS AFFECTING DRUG METABOLISM
• State of health
– Hepatitis
– Liver cancer
– Cardiac insufficiency
– Uremia
• degree of protein binding
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164. Changes In Drug Metabolism As A Consequence Of Hepatic Disease
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From Principles of Drug Action
166. FACTORS AFFECTING DRUG METABOLISM
• Gender
– Most studies are performed in the rat. In general,
male rats metabolize drugs faster than female rats
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168. FACTORS AFFECTING DRUG METABOLISM
• Degree of protein binding
– Conditions that displace bound drug from protein
allows more of the drug to be accessible to the
enzyme for which it serves as a substrate e.g.
uremia, low plasma albumin
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170. FACTORS AFFECTING DRUG METABOLISM
• Species variation
– Quantitative
– Qualitative
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171. Factors Affecting Drug Metabolism
• Species variation
– Human beings metabolize amphetamine by
deamination; rats and dogs metabolize the drug
by aromatic hydroxylation
– Guinea pigs have very little sulfotransferase
activity, humans have substantial activity
– Guinea pigs do not N-hydroxylate substrates;
mice, rabbits, dogs do
– Hexobarbital is metabolized at different rates by
different species
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174. Factors Affecting Drug Metabolism
• Substrate competition
– Two or more drugs competing for the same
enzyme can affect the metabolism of each other;
the substrate for which the enzyme has the
greater affinity would be preferentially metabolized
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