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)
CYP1A2 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

ADME ADME ISSUES IN ANTIssues I-CANCER DRUG DEVELOPMENT ADME

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

Drug Input & Different Routes of Administration

1. I.V. and I.A. injections:
2. 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

Plasma Water-3.5 L, ~4.5 % body wt (w/w) 100 mg 100 mg

Conc = 2 mg/ml Vd = 50 ml

Conc = 0.2 mg/ml Vd = 500 ml

T B W

E C W

Total extracellular water - 15 L, 20 % body wt (w/w) Total Intracellular water 20 L, 30 % body wt (w/w)

Beaker without Charcoal

Beaker with Charcoal

Total body Water 40 L, ~55 % body wt (w/w)

Major Drug Elimination Pathways (Coordinated defense mechanism)

Biotransformation

Excretion

HEPATIC

Extra-Hepatic

Renal

Biliary

Phase I

Phase II

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

CYP2C8 Paclitaxel Rosiglitazon e CYP2C9 cerivastatin(15%) Includes: warfarin phenytoin tolbutamide Losartan

CYP3A (50%)

Includes: lovastatin cyclosporin nifedipine midazolam CYP2D6 ethinylestradiol (25%) Ritonavir Includes: Tricyclic antidepressants, Midazolam SSRI's, haliperidol, propanolol, atomoxetine testosterone 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 Phenobarbital Taxol Phenylacetone Hydroxyphenobarbital 6-hydroxytaxol

Active Drug to Active Metabolite

Codeine Procainamide tamoxifen Morphine N-acetylprocainamide 4-hydroxytamoxifen

Drug Biotransformation Reactions

Inactive Drug to Active Metabolite
Hetacillin Sulfasalazine Cyclophosphamide Ampicillin Sulfapyridine + 5 ASA 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.)

CYP3A4
Family Subfamily 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 10t Cp Cp e

D Cp Vd

Two-compartment Open model

t Zt Cp C1 Cz
1
Central or Plasma
I.V. bolus

Tissue Dt Ct
Vt

Cp1 VC Dp

k12

k21

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 Blood Flow (ml/100 g tissue/min)
550 450

Adrenals Kidney Liver

Hepatic Portal

0.02 0.4 2.0

1 24 25 5 20

20 75 55 5 3 1 1

Brain Skin Muscle (basal) Connective Tissue Fat

2.0 7.0 40.0 7.0 15.0

15 5 15 1 2

Extravascular dose
e.v. dose

ka Site of absorption

Dp Cp Vd

k10
16
Cp'

12

Conc(ug/ml)

8
Cp Cp'-Cp

0 0 5 Time(hrs) 10 15

Cp=

F.Dose.Ka V(ka-k)

(e-k.t- e-ka.t)

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

AUC
Trapezoidal Rule

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

AUC

Example
Time (hr) 0 0.25 0.5 1 2 4 6 8 10 12 14 Conc (ug/ml) 0 2.025 3.53 6.07 8.75 9.36 8.1 6.41 5 3.71 2.75 AUC(0- ) AUC(ug.hr/ml)
10

0 0.25 0.69 2.40 7.41 18.11 17.46 14.51 11.41 8.71 6.46 19.38 106.80

Conc(ug/ml)

0 0 2 4 6 8 10 12 14 16

Tim e(hr)

Cp(last)= 2.75/0.1419
Conc Time Profile (Oral Dose)
10

y = 20.245e-0.1419x R2 = 0.9981

Conc (ug/ml)
1 0 2 4 6 8 Time (hr) 10

12

14

16

Bioavailability
Absolute Bioavailability
F= [AUC]e.v/[DOSE]e.v [AUC]i.v/[DOSE]i.v

Relative Bioavailability

F=

[AUC]e.v/[DOSE]e.v [AUC]std/[DOSE]std

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 Cpmax consideration Cpmin consideration Cpmax and Cpmin consideration

Practical Considerations in Decision Making

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 interval Half-Li

on

Goals of the Dosing Regimen

Dosing Regimen: Loading and Maintenance Doses

Constant Rate Regimens

Sources Sourcesof of Variability Variability

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
Anti-epileptic Drugs
Carbamazepine Phenobarbital Phenytoin Topiramate Felbamate

Anti-Infective Agents

Anti-Cancer Drugs

Miscellaneous
Lovastatin Troglitazone Omeprazole Prednisolone Probencid Phenylbutazone Diazepam fexofenadine Hyperforin

Rifampicin Paclitaxel Rifabutin Docetaxel Rifapentine Cyclophosphamide Clotrimazole Ifophosphamide Sulfadimidine Tamoxifen Suflinpyrazone 4-hydroxyEfavirenz tamoxifen Amprenavir SU5416 Nelfinavir Ritonavir Capravirine

Induction of CYP1A2 (Ethoxyresorufin O-deethylase) 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 Treatment Once weekly (n=3) Twice weekly (n=3) Daily dosing (n=3) AUC Day 8 156 117 329 187 AUC Day 15 131 140 117 92 AUC Day 21/22 141 90 198 321 Induction of clearance 10% 40%

412 111

21 36

9 16

98%

Stopeck et.al. Clin. Cancer Research, 2002 Salzberg et.al, Investigational New Drugs 24: 299304, 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

7
ID: 4

3
ID: 5

7 30

10
ID: 6 ID: 7 ID: 8 ID: 9

30

Midazolam Conc. (ng/ml)

10
ID: 10 ID: 11 ID: 12 ID: 14

30

10
ID: 15

30

10

Time(hrs)

Day 0 Day 1 Day 42

64

Inhibition of Drug Metabolizing Enzymes

Inhibitor absent Inhibitor present

CYP3A
Active drug
Active drug

CYP3A
Inactive drug

Inhibitor
Inactive drug

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

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 -

Former Student/Post-Doc
Srikanth Nallani, Ph.D., FDA

Funding Sources
- Aventis Pharmaceutical, Eli Lily & Co, Bristol Myers - Womens Health (UC), American Cancer Society - NIH, Susan G. Komen Breast Cancer Foundation Squibb