Key Pharmacokinetic

Concepts Single Dose and

Steady State Drug
Administration
Pankaj B. Desai. Ph.D.
Professor of Pharmacokinetics
and Biopharmaceutics
Director, Drug Development
Graduate Program

Morning Agenda: Wake Up and

Smell the Coffee (Cytochrome
P450 1A2 Substrate)
Overview of ADME principles
Important PK Parameters
First Pass Metabolism
Compartmental & NonCompartmental Analyses
Single Dose Kinetics
Multiple Dose Kinetics
Drug-Drug Interactions
Inter-Subject Variability

CYP1A2
Substrate

ADME ISSUES IN ADME

ANTIssues I-CANCER DRUG
ADME
DEVELOPMENT

Clinical Pharmacology
First in Human -Pharmacokinetically Guided Dose
Escalation/ Drug Tolerance Study
Pharmacokinetics-Pharmacodynamics
Drug Metabolism
Mass Balance with Radiolabeled Compounds
Bioequivalence:Generic compounds
Single and multiple doses
Conventional versus controlled release formulations
Bioavailability of metabolites

Drug-Drug/Drug Dietary Product Interactions

Special Populations

1.

2.

Drug Input & Different Routes of

I.V. and I.A. injections: Administration

Bolus dosing

Zero-Order Input (Infusions)

Extravascular Administration

First Order (mostly passive diffusion)

Zero Order (active transport and controlled release systems)

Factors Affecting Drug

Distribution
Phyisco-chemical properties of the drug
Small vs. Large mol.wt. Compounds
Hydrophilic vs. Lipophilic compounds
pH of the milieu and pKa of the drug

Perfusion rate (blood flow/min/g tissue)

Protein binding
Anatomical restrictions

CNS- protected by the blood brain barrier

Transport across placenta
Salivary Drug Excretion (S/P ratios)
Excretion of the drug in milk (M/P ratios)

Apparent Volume of Distribution

Mathematical term to correlate amount &
concentration
Merely a tool to understand the EXTENT of drug
distribution- not a real physiological volume
Compare to the volume of body waters
Best calculated from I.V. Dosing as
I.V. Dose/Cpo

Drug
Sulfisoxazole
Phenytoin
Phenobarbital
Diazepam
Digoxin

L/Kg
0.16
0.63
0.55
2.4
7

L/70 kg
11.2
44.1
38.5
168
490

Apparent Volume of Distribution

100 mg

Conc = 2 mg/ml
Vd = 50 ml

Beaker without
Charcoal

100 mg

Conc = 0.2 mg/ml

Vd = 500 ml

Beaker with
Charcoal

T
B
W

E
C
W

Plasma Water-3.5 L,
~4.5 % body wt (w/w)
Total extracellular
water - 15 L, 20 %
body wt (w/w)
Total Intracellular water
20 L, 30 % body wt
(w/w)

Total body Water 40 L,

~55 % body wt (w/w)

Major Drug Elimination Pathways

(Coordinated defense mechanism)

Biotransformation

HEPATIC

Phase I

Excretion

Extra-Hepatic
Renal

Phase II

Biliary

Glomerular Filtration
Kidney receives 1.1 L of blood (20 25%)
of cardiac output
10 % is filtered at the glomerulus
Compounds with Mol.wt < 20,000 filtered
GFR = 120 ml/min
CLR of Inulin - a measure of GFR
Filtered freely into the tubule
Not influenced by protein binding and neither
secreted nor reabsorbed

Rate of filtration = Fu. Cp.GFR

Not a very effective drug extraction
process (maximal ~ 0.11 or 10 %)

Active Secretion

Detected when the overall rate of urinary

drug excretion exceeds the rate of filtration
Secretory processes (proteins) located
predominantly within the proximal tubules
Mechanisms exist for secreting acids
(anions) and bases (cations) from plasma
into the tubular lumen
Energy-dependent
Saturable processes
Subject to competitive inhibition
Effect of Protein-Binding
Depends upon secretion efficiency
and contact time at the secretory
sites
Restrictive (dependent on the Fub)
vs. Non-Restrictive (perfusion-rate
limited)

Reabsorption
Must occur when CLR < fu.GFR
Reabsorption occurs all long the nephron,
associated with reabsorption of water; majority
however occurring from the proximal tubules
Predominantly a passive diffusion process
Driven by concentration-gradient across
the tubular lumen
Active secretion occurs for many
endogenous compounds such as vitamins,
electrolytes, glucose and amino acids
Urine-Plasma Ratio (U/P) based on HendersonHasselbalch equation
Influence of pKa and pH of urine

Major Tissues Involved in

Drug Metabolism

Liver
Small intestines
Kidney
Lung
Other portals of entry into the body
and protected organs.
-e.g. nasal mucosa

Representation of drug
metabolism and excretion by the
hepatocyte

Biliary Excretion is Transporter

Mediated

Phase I and Phase II

Drug Metabolizing
Enzymes

Phase I enzymes: Predominantly cytochrome P450 (CYP)

Drug Metabolism by
CYPs
Theophylline,
caffeine,
Olanzapine
CYP2C8
Paclitaxel
Rosiglitazon
e
CYP2C9
cerivastatin(15%)
Includes:
warfarin
phenytoin
tolbutamide
Losartan

CYP2A6 (Coumarin)
CYP2E1
CYP1A2(Chlorzoxazone)
CYP2B6
5%
bupropion,
tamoxifen,
efavirenz

CYP3A (50%)

Includes:
lovastatin
cyclosporin
nifedipine
midazolam
CYP2D6
ethinylestradiol
(25%)
Ritonavir
Includes: Tricyclic antidepressants,
Midazolam
SSRI's, haliperidol, propanolol, atomoxetinetestosterone
Detxromethorphan,

Phase II Reactions
Also known as Synthetic (conjugation)
reactions
Major reaction: Transfer of the conjugating
moiety to the drug
Enzymes involved are transferase

Glucuronosyl transferase
Sulfotransferases
N-acetyltransferase
Methyltransferase
Glycine transferase
Glutathione-S-transferase

Drug Biotransformation
Reactions

Active Drug to Inactive Metabolite

Amphetamine Phenylacetone
Phenobarbital
Hydroxyphenobarbital
Taxol
6-hydroxytaxol

Active Drug to Active Metabolite

Codeine Morphine
Procainamide N-acetylprocainamide
tamoxifen
4-hydroxytamoxifen

Drug Biotransformation
Reactions

Inactive Drug to Active Metabolite

Hetacillin Ampicillin
Sulfasalazine
5 ASA
Cyclophosphamide

Sulfapyridine +
Nitrogen mustard

Active Drug to Reactive Intermediates

Acetaminophen Reactive metabolites
(hepatic necrosis)
Benzo(a)pyrene
Reactive
metabolite
(carcinogenic)

Nomenclature
Basis: Amino acid sequence
Families: Less than 40 % a.a.
sequence
assigned to
different gene families
(gene
families 1, 2, 3, 4 etc.)
Subfamilies: 40 55 % identical
sequence
(2A, 2B, 2C, 3A etc.)
Family

Subfamily
CYP3A4

Isoform

CYP Nomenclature
(Contd.)
Cytochrome P450 Nomenclature, e.g.
for CYP2D6
CYP = cytochrome P450
2 = genetic family
D = genetic sub-family
6 = specific gene
NOTE that this nomenclature is
genetically based: it has NO
functional implication

Examples
of
CYP
mediated
Oxidative
Examples of reactions catalyzed by cytochrome P450:
Metabolism
Hydroxylation of aliphatic carbon

Examples of CYP mediated Oxidative

Heteroatom dealkylation
Metabolism
Examples of reactions catalyzed by cytochrome P450:

Clearance Concepts

Compartmental Modeling

One-Compartment
Open Model
I.V. bolus

DB1 Cp1
Vd
k10
K10 = overall
Elimination
Rate Constant

I.V. Bolus

k
10 t
Cp C p e

D
Cp
Vd

Two-compartment Open
model
1t
Zt
Cp C1
Cz
Central or Plasma
I.V. bolus

Cp1 VC
Dp

k12
k21

Tissue
Dt
Ct
Vt

1- hybrid rate constant (distribution)

z- hybrid rate constant (terminal)

Two-compartment Open Model

Elimination
only

Blood flow to human tissues

Tissue

Percent Body
Weight

Percent Cardiac
Output

Adrenals

0.02

550

Kidney

0.4

24

450

Liver

2.0

25

Hepatic
Portal

Blood Flow
(ml/100 g
tissue/min)

20

20

75

Brain

2.0

15

55

Skin

7.0

Muscle
(basal)

40.0

15

Connective
Tissue

7.0

Fat

15.0

Extravascular dose
e.v. dose

Site of ka
absorption

Dp
Cp
Vd

k10

NCA
Used to estimate
AUC
Bioavailability
Clearance
Volume of Distribution
Average Steady State
Concentration

AUC
Trapezoidal Rule

AUC= (t3-t2)
(C2+C3)

AUC

Example
10

Conc(ug/ml)

0
0

Tim e(hr)

Cp(last)=
2.75/0.1419

10

12

14

16

Bioavailability
Absolute
Bioavailability

Relative
Bioavailability

Bioequivalence
Two products are considered to be
bioequivalent if the concentration time
profiles are so similar that they are
likely to produce clinically relevant
differences in either efficacy or toxicity.
Common measures used to assess
differences are
Tmax, Cmax and AUC.

Other Parameters
CL = Di.v/AUC
AUMC = (t2-t1)(C1t1 +C2t2)
MRT (Mean Residence Time)
= AUMC/AUC
or MRT = 1/K or CL/V
Vss = CL. MRT

Multiple Dosing Overall Aims

Key Concepts
Principle of Superposition
Drug Accumulation and Steady State
Persistence Factor and Accumulation Factor
Peak, Trough and Steady State Average Levels

Applications
Determination of drug concentrations and amounts
following
multiple i.v. and e.v. doses (Ka > > K10)
max, min and during a dosing interval

Determination of dosing regimens

Doses (Maintenance and Loading) and Dosing Interval
C max consideration
p
C min consideration
p
C max and C min consideration
p
p

Drug Accumulation
Depends on Frequency of
Administration

Multiple I.V. Dosing

The AUC within a dosing interval at

steady state is equal to the total AUC
of a single dose.

Peak, Trough and Css

Average

Accumulation Index - Cssmax/Cmax1

AUC at Steady State = AUC0

Impact of Half-life and dosing interv

Half-Life on

Goals of the Dosing

Regimen

Dosing Regimen: Loading

and Maintenance Doses

Constant Rate Regimens

Sourcesof
of Variability
Variability
Sources

Genetic factors
Genetic differences
within population
Racial differences among
different populations
Environmental factors and
drug interactions
Enzyme induction
Enzyme inhibition
Physiologic considerations
Age
Gender
Diet/nutrition
Pathophysiology
Drug dosage regimen
Route of drug
administration
Dose dependent (nonlinear)
pharmacokinetics

Examples of CYP3A Inducers

Therapeutic Class
Antiepileptic
Drugs

AntiInfective
Agents

Anti-Cancer
Drugs

Miscellaneou
s

Carbamazepi Rifampicin
Paclitaxel
Lovastatin
ne
Rifabutin
Docetaxel
Troglitazone
Phenobarbital Rifapentine Cyclophospham Omeprazole
Phenytoin
Clotrimazole
ide
Prednisolone
Topiramate Sulfadimidin Ifophosphamide
Probencid
Felbamate
e
Tamoxifen
Phenylbutazo
Suflinpyrazo
4-hydroxyne
ne
tamoxifen
Diazepam
Efavirenz
SU5416
fexofenadine
Amprenavir
Hyperforin
Nelfinavir
Ritonavir
Capravirine

Induction of CYP1A2 (Ethoxyresorufin Odeethylase) by SU5416 in Primary Human

Hepatocytes

Stopeck et.al. Clin. Cancer Research, 2002

Salzberg et.al, Investigational New Drugs 24: 299304,
2006)

Example of Auto-Induction SU5416

Oral
Treatmen
t

AUC Day 8

AUC Day
15

AUC Day
21/22

Induction of
clearance

Once
weekly
(n=3)

156 117

131 140

141 90

10%

Twice
weekly
(n=3)

329 187

117 92

198 321

40%

Daily
dosing
(n=3)

412 111

21 36

9 16

98%

Stopeck et.al. Clin. Cancer Research, 2002

Salzberg et.al, Investigational New Drugs 24: 299

304, 2006)

Effect of Tamoxifen (TAM) Mediated

CYP3A4 Induction
Letrozole Alone

Letrozole + Tamoxifen
( 6 weeks & > 4 months)

Dowsett, M. et al. Clin Cancer Res 1999;5:2338-2343

62

PXR

Pharmacogenomics. 2008 November; 9(11): 16951709.

Midazolam
Plasma
Conc.
Profile
Effect
of CYP3A/PXR
Genotypes
on CYP3A
Induction
3
ID: 1

ID: 3

3
ID: 4

ID: 5

30
10

Midazolam Conc. (ng/ml)

30

ID: 6

ID: 7

ID: 8

ID: 9

ID: 10

ID: 11

ID: 12

ID: 14

10
30
10
30

ID: 15

Time(hrs)

10

Day 0
Day 1
Day 42
64

Inhibition of Drug Metabolizing

Enzymes
Inhibitor present

Inhibitor absent

CYP3A

CYP3A
Active drug

Active drug

Inactive drug
Inactive drug

Inhibitor

Saquinavir
+
Ritonavir

AIDS. 1997 Mar

15;11(4):F29-33

Saquinavir

Plasma Rosuvastatin concentration-time

profile in the absence and presence of
Darunavir/Ritonavir

Before DRV/RTV

After DRV/RTV

Desai Lab with the UC President

Graduate Students
- Rucha Sane
Niresh Hariparsad
Fang Li
Ganesh Mugundu

Former Student/PostDoc Srikanth Nallani, Ph.D., FDA

Collaborators

Arthur Buckley, Ph.D., College

of Pharmacy
Julie Nelson, Ph.D., Department
of Molecular Genetics,
Biochemistry and Microbiology
- Elizabeth Shaughnessy, MD
- Judith Feinberg, MD Brian
Goodwin, Ph.D.,
GlaxoSmithKline
Stephen Storm, Ph.D.
University of Pittsburgh
-

Funding Sources
- Aventis Pharmaceutical, Eli Lily & Co, Bristol Myers
Squibb
-

Womens Health (UC), American Cancer Society

NIH, Susan G. Komen Breast Cancer Foundation