Drug – Drug Interaction and Drug Transporters :

What ‘ s new ?

J.M.SCHERRMANN

Club Phase 1 Meeting, Paris, 22 March 2016

FROM WHERE DO WE COME?

TRANSPORTERS and PHARMACOKINETICS in 1987Michael SCHWENK : Review, Drug Transport in intestine, liver and kidney.

Arch. Toxicol. 60, 37, 1987

"Drug transport in the intestine

occurs mainly by diffusion"....

" Intestinal drug carriers

contribution seems to be of minor

importance."

"The liver is the organ where

carrier-mediated drug transport

predominates"

ENZYMES and PHARMACOKINETICS

in 1987

« Genetic polymorphism of human

cytochrome P450 (S)-mephenytoin 4-

hydroxylase (P-450 meph)

(U.T. MEIER and U.A. MEYER, 1987) »

« Metabolism of cyclosporin A. Interaction

of erythromycin, using rabbit hepatocytes

and microsome fractions

(P-450 LM3c or P-450IIIA4)

I. FABRE et al. (J.P. CANO), 1988

TRANSPORTERS and PHARMACOKINETICS

in 1987Michael SCHWENK : Review, Drug Transport in intestine, liver and

kidney. Arch. Toxicol. 60, 37, 1987

Four hypothetical carrier systems for

hepatocellular drug uptake

Carrier 1 Carrier 2 Carrier3 Carrier4

Bile acids

Estronesulfate

Estradiol

glucuronide

Bilirubin

Bromosulphophtalein

ANS

Indocyanine green

Morphine

NalorphineOuabain

"bile acid carrier" "organic anions" "organic cations"

OATPs

NTCP

OATPs

NTCP

OCT1, 3 OATPs

DIFFUSIONAL versus VECTORIAL PHARMACOKINETICS

X

CYPs

CEs

X-OC

X

X-OH

X-OC

D

A

E

Ante 2000

« Diffusional PK »

Passive

Diffusion

Passive

Diffusion?

M

XXX

CYPs

CEsX-OH

Post 2000

« Vectorial PK »

Active

Efflux

Active

Influx

Active

Efflux

Passive

Diffusion

X

Phase 0

Phase I

Phase II

Phase III

X-OC

X-OC

1st period : cancer research (1970 - )

MDR (Multi Drug Resistance) phenotype

( Juliano and Ling , 1976 )

MDR1, P-glycoprotein, ABCB1

Mdr1a et Mdr1b in rodentscavéoline

2nd period : pharmacology (1990 - )

Expression in healthy tissues (Protection and

Detoxification , the << GATEKEEPER >> )

1st PK-Pgp study published by Tsuji and Terasaki in

1992 (Life Sci.51, 1427,1992 )

3rd period : Guidance for Industry (2006 - )

substrate/inhibitor identification

P-GLYCOPROTEIN : PIONEER and LEADER

Aller et al., Science

2009,323:1718‐1722.

70 Å

~13

6 Å

~30

Å

WHERE WE ARE TODAY ?

• ABC (ATP Binding Cassette) SUPERFAMILY48 human genes; ≈ 9 drug transporters

SUPERFAMILIES and FAMILIES of DRUG TRANSPORTERS

• SLC (SoLute Carrier) SUPERFAMILY

> 362 mammalian genes; ≈ 30 drug transporters

« 15% of human genes code for 4500 transport proteins »

Paulsen, FEBS Lett., 1998 ; J. Mol. Biol., 1998

EFFLUX (Conjugated Metabolites)

EFFLUX (Xenobiotics(MDR, Tissue Defense)

SECONDARY (TERTIARY) ACTIVE TRANSPORTERS

ABC Superfamily

PRIMARY ACTIVE TRANSPORTERS

S

ATP

S

ATP

S ABCCS

ABCB, ABCG

GSH

INFLUX and/or EFFLUX(Xenobiotics, Conjugated Metabolites)

SLC Superfamily

1

2

S S

3

H+

H+

K+

Na+

ABC and SLC TRANSPORTERS

P-glycoprotein, ABCB1, MDR1

Breast Cancer Resistance Protein (BCRP, ABCG2)

MRPs, Multi-Drug resistance associated Proteins

OATs Organic Anion T.OATPs Organic Anion Polypeptide T.OCTs Organic Cation T.

Membrane Transporters

in Drug Development:

Giacomini et al., Nature Rev

Drug Disc 2010

White Paper 2010

ABC Transporters

P-gp

BCRP

BSEP

MRP2

MRP3

MRP4

MDR3

International Transporter Consortium in 2007

White Paper 2010

SLC TransportersOATP1B1

OAT1

OATP1B3

OAT3

OCT2

OATP1A2

OATP2B1

OCT1

PEPT1

PEPT2

MATE1

MATE2

Membrane Transporters

in Drug Development:

Giacomini et al., Nature Rev

Drug Disc 2010

P-gp/BCRP Inhibition Tree

High Solubility Low Solubility

HighPermeability

Class 1(High Solubility, High

Permeability)Metabolism

”High hepatic extraction”Transporter effects

minimal

Class 2(Low Solubility, High

Permeability)

MetabolismEfflux transporter effects

predominate

LowPermeability

Class 3(High Solubility, Low

Permeability)

Renal and/or BiliaryElimination

Absorptive transporter effects predominate

Class 4(Low Solubility, Low

Permeability)

Renal and/or BiliaryElimination

Absorptive and efflux transporter effects could be

important

The Biopharmaceutics Classification System and transporters

From G. Amidon , Pharm. Res, 12, 413, 1995 and C.Y. Wu , Pharm Res, 22, 11, 2005

INTESTINE(enterocyte)

FECESORALEXPOSURE SecretionAbsorption

LIVER(hepatocyte)

BILE

Efflux

Uptake

Excretion

BBB(brain microcapillary

endothelial cell)

Efflux Uptake

BRAIN

KIDNEY(tubuleproximal cell)

URINE

Secretion

Reabsorption

Filtration

From JM SCHERRMANN-Comprehensive Med.Chem, 2007

PHARMACOKINETIC ROLE OF TRANSPORTERS

Drugs as substrate, inhibitor or regulator?

TRANSPORTER PROTEIN

INHIBITION

MDR Reversal

TRANSPORT

SUBSTRATE

EXPRESSION

( Induction or Repression of Regulation PathwaysGenetic Polymorphisms)

And Candidates for DDI ?

INTESTINAL LUMEN

SYSTEMIC BLOOD

;;

basolateral

membrane

Apical

(brush border)

membrane

MRP5 MRP4 MRP3 OCT1 MCT1

MRP2 BCRP MDR1 OATP1A2

OATP2B1

OATP3A1

OATP4A1

MCT1PEPT1

IMPACT OF TRANSPORTERS ON DRUG ABSORPTION

FRUIT JUICE, A GLASS FULL OF DRUG INTERACTIONS

F Fexofenadine (57 %), Talinolol (44 %), Celiprolol (87 %)

CYP3A4

OATP1A2

ABCB1

!

IC50 5 M

IC50 3000 M

GRAPE FRUIT JUICE

Naringin

ORANGE FRUIT JUICE

Hesperidin

COMPETITIVE

INHIBITION GRAPE FRUIT JUICE

Bergamottin6,7-dihydroxybergamottin

MECHANISM-BASED

INHIBITION

From DG Bailey et al, CPT, 81, 2007

BIOAVAILABILITY OF FEXOFENADINE

Influence of delay betweengrapefruit juice and fexofenadine intake

4h

+ 2h (37%)

0h (57%)

F

From H. Glaeser, Clin. Pharmacol. Ther., 2007

Traditional Chinese Medicine and Transport Interactions

Herb Active compounds Transporter target DDI risks Ref

Rhubarb AnthraquinonesOAT1OAT3

(kidney)

YesInhibition

Ma LJ.Ethnopharmacol

2014

Turmeric (Curcuma longa)

Curcumin

ABCA1ABCB1ABCC1ABCG2

Inhibition (MDR

reversal)

Zhang X Front. Physiol 2014

DanshenRosmarinic acid

Tanshinol Lithospermic acid

OAT1OAT3

(kidney)

Yes Inhibition

Wang L Drug Metab

Pharmacokinet. 2013

Huang-Qin-Tang MultipleMCT1

(intestine and BBB)

Yes Inhibition

Yu CP Phytomedicine

2013

Yanhusuo (YHS) glaucineABCB1ABCC1

Yes Inhibition

(MDR reversal)

Lei Y Food Chem 2013

BLOOD

IMPACT OF TRANSPORTERS ON DRUG AT THE HEPATO-BILIARY

LEVEL

From JM SCHERRMANN-Comprehensive Med.Chem, 2007

UPTAKE

Phase 0

EFFLUX

Phase 3a

EFFLUX

Phase 3b

DRUG INTERACTIONS MEDIATED BY TRANSPORTERS

From W. Muck, Clin Pharmacol Ther, 65, 251, 1999

CERIVASTATIN (0.2 mg/d)-CYCLOSPORINE (400mg/d)

in kidney transplant recipients

Myopathy and Rhabdomyolysis

AUC X 5

SIROLIMUS AUCX3

TACROLIMUS AUCX 1,3

PgpCsA

CsA (Ki≈0,2µM)

Cerivastatin

OATP1B1

CsA

(IC50 ≈30µM)

CYP3A bile

SNP

c521T>C

Atorvastatin

OATP1B1

Link et al, NEJM 2010

Effect

Rhabdomyolysis (OR 16)

HEPATIC UPTAKE AND BILIARY EXCRETION OF

VALSARTAN

(angiotensin II AT1-receptor antagonist)Valsartan

Absorption (F ≈ 30-50%)Faecal excretion ≈ 85% (bile)Vss ≈ 17 LClt ≈ 2,2 L/ht1/2 ≈ 7h

From W. Yamashiro et al, Drug Metab Dispo. 2006

UPTAKE

Phase 0

EFFLUX

Phase 3a

BCRPMDR1 MRP4-5

OATP1A2URAT1

OCTN2

MRP1-2-3-5-6 ?OAT3

BLOOD

BRAIN

luminal

membrane

abluminal

membrane

tight

junctions

Transporter-mediated Drug Interactions at BBBIMPACT OF TRANSPORTERS ON DRUG DISTRIBUTION

BLOOD-BRAIN BARRIER

MCT1

MCT1

P-Glycoprotein and Breast Cancer Resistance Protein :

Two Dominant Transport Working Together in

Limiting the Brain Penetration of Topotecan

N.A. de Vries, Clin Cancer Res, 2007

3.20.652.0Brain/

Plasma

1.61.5AUC

Brain/WT

3.72.40.75AUC

Plasma/WT

Bcrp1(-/-)

Mdr 1a/1b (-/-)

Bcrp1(-/-)Mdr1a/1b(-/-)

12

Topotecan IV 5 mg/kg

« Synergistic Effect of P-gp and BCRP

on common substrates »

Erlotinib, Gefitinib, Lapatinib, Imatinib

Flavopiridol, Mitoxantrone, Prazosin

IMAGING P-GLYCOPROTEIN TRANSPORT ACTIVITY AT THE HUMAN BBB WITH PET

Magnetic

resonance image

11C-Verapamil

Control + CSA

L. Sasongko et al,

CPT, 2005

J. Bart, NeuroImage, 2003

CSA

50µg/kg

0

Basolateral

membrane

OCTN1- 2

OAT2

Apical

membrane

URAT1

OAT4

OATP4C1

OAT 1- 3

MRP1- 3- 5- 6

MDR1, BCRP

MRP2- 4

OCT 1-2- 3

PEPT1- 2

SAT1OATP1A2

IMPACT OF TRANSPORTERS ON DRUG ELIMINATION

RENAL LEVEL

From JM SCHERRMANN-Comprehensive Med.Chem, 2007

BLOOD URINE

MATE1

V ≈ 25 L

fup = 0,97

t1/2 6- 10h

Clr (# 100Clt)

Clr =250 ml/min

activité

extracellulaire

Nature 438, 3 novembre 2005

Phosphate

d’Oseltamivir

(Tamiflu®)

Estérases Carboxylate

d’Oseltamivirneuraminidases

(virus grippal)

PK PD

Wartime tactic doubles power of scarce bird-flu drug

Nature 438, 3 novembre 2005

Transporter-mediated Drug Interactions: OAT1, SLC22A1

Renal tubules

OAT1

Wartime tactic doubles power of scarce bird-flu drug

PBN

Transporter-mediated Drug Interactions at Kidney

From Koepsell and Endou, Eur J Physiol, 447, 666, 2004

DDI : Salicylate-methotrexate

(1000 mg) (2.5 mg)

MATE1

Paraquat, 1-methyl-4-phenyl

pyridinium (MPP+)

Lee W.K, Curr Drug Metab 2009

Pralidoxime, (Antidote of

organophosphate)

Renal secretion mediated by OCT1/2

Kayouka M, Crit Care Med, 2011

QUESTIONS ?

(I) In vitro - in vivo correlation

R = fold systemic AUC change (≈ CYP-based drug interactions)

Cl

Cl

AUC(inhibitor) 1R= = 1

fAUC

fu I1+ +1-f

Ki

Ki = in vitro inhibition constant

[I] = inhibitor plasma concentration

fu = unbound plasma fraction

fcl = fraction of the total clearance mediated by the affected transporter

Cl

Iif f =1 R=1+fu (2)

Ki

Question : Quantitative Prediction of in vivo Drug Interaction?

DRUG INTERACTIONS MEDIATED BY TRANSPORTERS

(I) In vitro - in vivo correlation

From CJ. Endres, Eur J Pharm Sci 27, 501, 2006

IAUCR=

AUC

fcl

DRUG INTERACTIONS MEDIATED BY TRANSPORTERS

(II) Quantitative prediction of in vivo drug interaction

From C.J. Endres, Eur J Pharm Sci, 27, 501, 2006

K.Ball et al., AAPS J., 2013

Question : Interspecies differences in Transporter Expression

ABC and SLC differences between species in brain microvessels

ABC differences between in vivo / in vitro BBB (hCMEC/D3)

MDR1

BCRP

MRP1

MRP4

MRP5

0.001

0.01

0.1

1

10

100

Relative expressionin hCMEC/D3compared to

human brain microvessels

S.Dauchy et al.,Biochem. Pharmacol., 2009

Relative to human

brain microvessels

Question : Are in vitro models reproducing the in vivo conditions

P-gp Efflux Ratio for Opioids

N. Tournier, Curr Pharm Design, 2011

11.3

Question : role of active metabolites ?

• Impact of Quantitative Proteomic MS-MS

on transportome atlas -the Terasaki Dream –

• From single transfected cell models to quadruple transfectedsystems – J. König –

• Computational prospecting for drug-transporter Interactions

• Development of non-invasive technologies including imaging

• Ontogeny in the expression and localization of transporters indifferent tissues

• Establish clinically important transporter polymorphisms

• Intracellular concentrations: measurement modeling implications for the liver

Questions : Many others……

WHERE IN ADME ?

TM

A

D E

EFFECT

PD

« Transport and Metabolism are Effectors of the time course of

Drug Absorption, Distribution, and Elimination »

FROM ADME TO ADE ?

THE ADE-TM triangles :

F

CltVd

JM Scherrmann, Chem. Biodiv. 6 (2009)