corticosteroid effects › assets › legacy_acops › 2008acop › main… · william j. jusko,...

William J. Jusko, Ph.D.Department of Pharmaceutical Sciences

ACoP 03/10/08

Corticosteroid Pharmacological

Effects

⇒ Treatment for Immune Related Diseases- Rheumatoid arthritis - Lupus erythematosus - Bronchial asthma- Organ transplantation

• Immunological Effects– Immunosuppressive– Anti-inflammatory

• Metabolic Effects– Carbohydrate metabolism– Lipid metabolism– Protein metabolism

Adverse Effects⇒ steroid diabetes⇒ abnormal fat distribution⇒ muscle wasting

negative nitrogen balance

Capacity-Limitation Turnover and Homeostasis

Effe

ct, %

Concentration

γγ50

γmax

CECCEE

+•

=

Hill Function

The Law of Mass Action( D + R DR ) and smallquantity of targets leads to capacity-limitations in most responses.

Production LossBiologicalFactor

(R)

RkkdtdR

lossproduction •−=

Both diseases and therapeuticagents often interfere with thehomeostasis in the body resulting from the natural turnover of biological substances or functions.

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Giant Rat Experiments: Single- dosing and chronic infusion regimens. PK assessment.Parallel Analysis of Multiple Tissues: Liver, muscle, kidney, fat, etc.Molecular Biology: Gene Arrays and QRTPCR of selected genes.Biochemistry: Measurement of relevant gene products (proteins and enzymes), and signals.Systems Physiology: Blood glucose, insulin, lipid profiles, others…‘Disease’ Models: Diabetes, Arthritis, Pregnancy.Development of Mechanism-Based PK/PD/PG/DISModels. Dhahbi et al, American Journal of Physiology, E352-60 (1999).

Metabolic/Genomic Effects of Corticosteroids

TyrosineAminotransferase

(TAT)

Catabolic Anabolic

Pharmacogenomics: Drugs & Genes & Models

Protein

Drug

DNA

Production Loss

Receptor

±

Production Loss

+

+

R. Ramakrishnan, DC Debois, RR Almon, NA Pyszczynski, and WJ Jusko, J. Pharmacokin. Pharmacodyn. 21: 1-24 (2002).

Fifth-Generation Model for Corticosteroid Pharmacodynamics: Application to Steady-State Receptor Down-Regulation and Enzyme Induction Patterns during Seven-Day Continuous Infusion of Methylprednisolone in Rats

Giant Rat Study

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Methylprednisolone PK/PD/PG in Rats

0 1 2 3 4 5 6 7

10000

1000

100

10

1MPL

Con

cent

ratio

n (n

g/m

l)

Time (hr)

0 4 8 12 16 20 24

DR

(N)

Con

cent

ratio

n(fm

ol/m

g pr

otei

n)

0

100

200

300

400

Time (hr)

0 12 24 36 48 60 72

GR

den

sity

(fm

ole/

mg

prot

ein)

0

200

400

600

800

1000

0 12 24 36 48 60 72

GR

mR

NA

(fm

ole/

gliv

er)

0

10

20

30

40

50

0 4 8 12 16 20 24

TAT

mR

NA

(pm

ole/

gliv

er)

0

1

2

3

4

Time (hr)

0 4 8 12 16 20 24

TAT

Act

ivity

(Δ

A/m

in/m

g pr

otei

n)0

1

2

3

PK GR mRNA TAT mRNA

DR(N) Cytosol GR TAT

Liver WeightSLR

ksLR kdLR

kdeg T

mRNATAT TATS

kdeg

EF

ksynD

DR

kd R (1-Rf) *kre

Rf*kre

kon kT

ks R

mRNARks Rm kd Rm

I(t)

RDR(N)+

Corticosteroid Pharmacogenomics

Fifth-generation modelRamakrishnan et alJPP 21: 1 (2002).

Model EquationsPHARMACOKINETICS tt

P eCeCCD ⋅−⋅− ⋅+⋅== 2121"" λλ

RECEPTOR DYNAMICS

RdRmRm

sRmR mRNAk

NDRICNDRk

dtdmRNA

⋅−⎟⎟⎠

⎞⎜⎜⎝

⎛+

−⋅=)(

)(150

LRmRNAmRNA obsR = 0R

sRmdRm mRNA

kk =

RkRDkNDRkRmRNAkdtdR

dRonrefRsR ⋅−⋅⋅−⋅⋅+⋅= )(

DRkRDkdt

dDRTon ⋅−⋅⋅=

)()( NDRkDRkdt

NdDRreT ⋅−⋅=

dRR

sR kmRNA

Rk ⋅⎟⎟⎠

⎞⎜⎜⎝

⎛= 0

0

Model EquationsLIVER WEIGHT RATIO

LRkNDRSkdt

dLRdLRLRsLR ⋅−⋅+⋅= ))(1(

mRNA DYNAMICS

mRNAkNDRSkdt

dmRNAPsyn ⋅−⋅+⋅= deg))(1(

LRmRNAmRNA obs= 0deg mRNAkksyn ⋅=

TAT DYNAMICS

TATkmRNASkdt

dTATTTATsT ⋅−⋅+⋅= deg)1(

LRTATTAT obs= 0

0

deg mRNATATkk TsT ⋅=

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Gene Arrays

0 12 24 36 48 60 72

GR

mR

NA

(fm

ole/

gliv

er)

0

10

20

30

40

50GR mRNA

Time (hr)

GENE TREE: 4373 of 8799 Probe Sets: Filtered based on genes expressed in liver.

EISEN

PLOT

www.sigenetics.comSilicon Genetics Inc.

Pattern Searches:User-Defined ProfilesSelf-Organizing MapsK-Means Clustering

7.0

Filtration System:Low Variability4 ti > 1.54 ti < 0.65

PK/PD Modeling (Adapt II)

Affymetrix Rat Gene Microarray Patterns: 192 / 8799 Genes

Jin JY, Almon RR, Dubois DC, Jusko WJ, JPET 307: 93-109 (2003).

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Extracting Global Systems Dynamics of Corticosteroid Genomic Effects in Rat Liver

E Yang, RR Almon, DC DuBois, WJ Jusko, IP Androulakis, JPET, in press (2008).

Genes ‘hashing’ to same integer belong to same cluster. n = 529 genes

Clusters of dynamic response patterns in tissues suggest that a limited array of control processes account for pharmacogenomic effects of steroids.

Baseline versus Chronic versus Acute dosing reveals myriad complexities in gene homeostasis.

Response profiles and PK/PD models offer opportunities to formulate hypotheses regarding factors and mechanisms of genomic effects.

Data available at: http://pepr.cnmcresearch.org/

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• Male Rats• Age : 6-9 weeks• Weight: Matched to ~175 g• Induce Arthritis & sacrifice at various times (days

9,12,15,17,19,21,23,25,30,34) up to day 34: for collection of paw tissue, plasma.

• Non-invasive Disease Endpoints: Paw Edema, Body Weight, Bone Mineral Density

• Various dosing regimens of dexamethasone.

Rats with CollagenInduced Arthritis

Justin C. Earp: Trying to gain their trust.

Time (hours post induction)

0 200 400 600 800 1000

2

50

75

100

125

150

175 Arthritic RatsHealthy Controls

Rheumatoid Arthritis: Disease Pathology• Immune Cells

– Migration – Proliferation

• Cyto/Chemo-kines: TNF-α, IL-1β, IL-6, IFN-γ, GM-CSF

• Prostaglandins• Nitric Oxide • Rheumatoid Factor• Edema• Joint Tissue

Erosion:– Bone– Cartilage– Synovial Tissue

Choy HS, et al. (2001) NEJM.

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Arthritic


0.1

1

10

100

1000

10000

10000 200 400 600 8000 200 400 600 800 1000

TNF-αIL-1β

IL-6

Healthy

Neeck G, Renkawitz R, Eggert M 2002.

Cytokines Cell Mol Ther7(2):61-69.

Paw Glucocorticoid Receptor mRNA & Corticosterone in Plasma


0 200 400 600 800 1000

GR

mR

NA

(ng_

mR

NA

/mg_

Tot

al_R

NA

)

0

100

200

300

400

500

600

700

0 200 400 600 800 1000

Cor

ticos

tero

ne C

once

ntra

tion

(ng/

mL

)

0

100

200

300

400

500

600

700

GR mRNA Corticosterone

Bone Mineral Density by Dual-Energy X-Ray Absorptiometry (Piximus, GE)

Total Femur

Diaphyseal Femur

Metaphyseal Femur

Epiphyseal Femur

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Bone Mineral Density: Femur & Lumbar

0.125

0.150

0.175

0.200

0.225

Bon

e M

iner

al D

ensi

ty (g

/cm

2 )

0.125

0.150

0.175

0.200

0.225

0 200 400 600 8000.075

0.100

0.125

0.150

0.175


0 200 400 600 8000.075

0.100

0.125

0.150

0.175

Diaphyseal Femur

Epiphyseal Femur

Metaphyseal Femur

Lumbar Vertebrate

Open: Controls; Closed: RA

0 4 8 12 16 20 241

10

100

1000

10000

TIME (hr)

0 4 8 12 16 20 24

IM Dose CT

VT

CP

VP

kA

CLD

CL

(F)

Healthy Rats Arthritic Rats

Doses: 2.25 and 0.225 mg/kg

Time (hours post induction)200 400 600 800 1000

IL-6

mR

NA

(ng_

mR

NA

/mg_

Tot

al_R

NA

)

0

2000

4000

6000

8000

10000

Time (hours post induction)200 300 400 500 600 700 800

IL-1

βm

RN

A(n

g_m

RN

A/m

g_T

otal

_RN

A)

0

50

100

150

200

High Dose: 2.25 mg/kgLow Dose: 0.225 mg/kg Disease Progression

Time (hours post induction)400 450 500 550 600 650 700

TN

F-α

mR

NA

(ng_

mR

NA

/mg_

Tot

al_R

NA

)

0

50

100

150

200

250

DEX PD: Paw Glucocorticoid

Receptor mRNA

DEX PD: Plasma Corticosterone


200 300 400 500 600 700 800C

ortic

oste

rone

(ng/

mL

)0

100

200

300

400


200 400 600 800 1000

GR

mR

NA

(ng_

mR

NA

/mg_

Tot

al_R

NA

)

100

200

300

400

500

600

700

Disease ProgressionHigh Dose: 2.25 mg/kg Low Dose: 0.225 mg/kg

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Acute & Chronic DEX PD: Paw Edema


200 400 600 800 1000

2

75

100

125

150

175Chronic: 0.225 mg/kgDisease Progression

Acute: 2.25 mg/kg Acute: 0.225 mg/kg


0 200 400 600 800

2

0.125

0.150

0.175

0.200

0.225

Chronic DEX PD: Bone Mineral Density

Healthy Rats: ο 0.045 mg/kg, once daily, 7 days (4 rats)Δ 0.225 mg/kg, once daily, 7 days (4 rats)

Arthrtic Rats: • 0.225 mg/kg, once daily, 7 days (6 rats)

Total Femur

DEX RA PD Systems Model

• Dex pharmacokinetics.

• Adrenal suppression by Dex.

• 5th-Gen Receptor/Gene control (Dex, CST).

• RA up-regulation of cytokine mRNA.

• Transductional control of BMD.

• Joint cytokine production of edema.

• Indirect response models for multi-component interactions.

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• Assessment of the kinetics and responses of steroids have offered numerous insights intomechanism-based PK/PD/Disease models.

• Drug-alterations of biological systems help probe underlying control steps.

• Models can integrate PK, receptor, gene, physiology, and disease processes.

• Models help formulate or alter hypotheses and design new experiments.

CollaboratorsRichard R. Almon, PhDDebra C. DuBois, PhDIoannis Androulakis, PhDEric Hoffman, PhD

NIH GrantsGM-24211GM-57980GM-67650

PhD Students (Recent)Yu-Nien (Tom) Sun, PhDRohini Ramakrishnan, PhDDonald E. Mager, PhDMahesh Samtani, PhDAna Hazra, PhDZhenling Yao, PhDEric Yang, PhDJustin Earp, PhD

and many previous fellows

and students.

TechniciansNancy A. PyszczynskiSuzette M. Mis

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corticosteroid effects › assets › legacy_acops › 2008acop › main… · william j. jusko,...

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