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First Analysis of Relation Between

CYP2C19 Genotype and Platelet Function in

Ticagrelor Versus Clopidogrel Treated Patients:

::

Results from the ONSET/OFFSET and RESPOND Studies









Gurbel PA1, Bliden K1, Storey RF2, Wei C3, Armstrong M3, Butler K3, Tantry US1



1Sinai Center for Thrombosis Research, Baltimore, Maryland, USA

2Univ of Sheffield, Sheffield, United Kingdom

3AstraZeneca, Wilmington, Delaware, USA

Disclosures



Research Grants/Support Honoraria/Consulting

Pozen Pozen

Astra Zeneca Novartis

Nanosphere Bayer

Accumetrics Astra Zeneca

Helena Eli Lilly

Accumetrics

Multiplate

Nanosphere

Portola

Sanofi Aventis

Daiichi Boehringer

Merck

Medtronic

Background

• Variable and insufficient active metabolite generation by hepatic CYP P450 isoenzymes

result in clopidogrel response variability and resistance.1

• LOF SNPs of CYP isoenzyme genes, particularly 2C19, linked to:

- diminished clopidogrel PK and PD effects

- increased ischemic events in PCI patients

- highlighted by FDA, EMEA, ACCF/AHA.1





• Ticagrelor is a reversibly-binding, noncompetitive, direct-acting, oral P2Y12 antagonist.2



• Ticagrelor therapy was associated with:

- less MACE in ACS patients in PLATO trial.3

- more rapid onset, offset, and greater level of inhibition in ONSET/OFFSET Study.4

- greater platelet inhibition in both clopidogrel responders and non-responders in

RESPOND Study.5

• Unknown facts:

- influence of CYP2C19 genotype on ticagrelor PD

- comparative PD effects of clopidogrel and ticagrelor in relation to 2C19 genotype

- whether ticagrelor treated patients have greater inhibition than clopidogrel treated

patients with “clopidogrel responder” genotype.





1. Bonello et al. J Am Coll Cardiol. 2010;56:1024-31. 2. Gurbel PA et al. Expert Opin Pharmacother. 2010;11:2251-9.

3. Wallentin L et al. N Engl J Med. 2009;361:1045-57. 4. Gurbel PA et al. Circulation. 2009;120:2577-85.

5. Gurbel PA et al. Circulation. 2010;121:1188-99.

Objectives

In stable CAD patients taking either ticagrelor or clopidogrel



in the ONSET/OFFSET and RESPOND studies:





1) compare platelet function between genetic categories

(CYP2C19 and ABCB1)

for each treatment:

Kruskal-Wallis test





2) compare platelet function between treatment groups

for each genetic category:

Wilcoxon rank-sum test

ONSET/OFFSET Study Design

Visit 1

Screening/Washout Period (n=154)

days

- 21

Screen Failures (n=31)



Visits 2-3

Onset









Randomization (n=123)

Platelet Function Testing

(Pre-dose and 0.5,1,2,4,8,24 hours post-dose)

1 day









180 mg Ticagrelor (n=57)

600 mg Clopidogrel (n=54) Placebo (n=12)

PM: 90 mg Ticagrelor

Maintenance



6 weeks ± 3

days









90 mg Ticagrelor bid 75 mg Clopidogrel qd Placebo







Visit 4

Offset









Last Dose

Platelet Function Testing

10 days









(0,2, 4,8 hours post last dose)





Visits 5-10

Platelet Function Testing

Day 1-3,5, 7 and 10 after last dose

RESPOND Study Design: Nonresponders





Screening * Randomization Period 1 Crossover Period 2

14 ± 2 days 14 ± 2 days

(n=144)



Treatment C (n=20) Treatment T (n=17)

Clopidogrel 600 mg/75 mg Ticagrelor 180mg/90 mg





Nonresponders

(n=41) Treatment T (n=21) Treatment C (n=17)

Ticagrelor 180mg/90 mg Clopidogrel 600 mg/75 mg









Visit 1 Visit 2 Visit 3 Visit 4= Visit 5

Visit 3+1 day

PK/PD: PK/PD: PK/PD: PK/PD:

Predose, Predose, Predose, Predose,

0.5,1,2,4,8 2,4,8 hours 0.5,1,2,4,8 2,4,8 hours

hours postdose hours postdose

postdose postdose



*patients were dosed with study drug >14 days after screening

RESPOND Study Design: Responders



Screening * Randomization Period 1 Period 2

14 ± 2 days 14 ± 2 days

(n=144)



Treatment C (n=13)

Clopidogrel 75 mg

Treatment C (n=29)

Clopidogrel 600 mg/75 mg

Treatment T (n=15)

Ticagrelor 180mg/90 mg



Responders Treatment C (n=13)

(n=57) Clopidogrel 600 mg/75 mg



Treatment T (n=28)

Ticagrelor 180mg/90 mg

Treatment T (n=13)

Ticagrelor 90 mg







Visit 1 Visit 2 Visit 3 Visit 4= Visit 5

Visit 3+1 day

PK/PD: PK/PD: PK/PD: PK/PD:

Predose, Predose, Predose, Predose,

0.5,1,2,4,8 2,4,8 hours 0.5,1,2,4,8 2,4,8 hours

hours hours

postdose postdose

postdose

postdose





*patients were dosed with study drug >14 days after screening

Demographics

Methods

Platelet Function Assessments:



- Light Transmittance Aggregometry (5 and 20 M ADP, maximal)

(Chronolog, Havertown, PA)



- VASP-P Assay (Biocytex, Marseille, FR)



- VerifyNow P2Y12 Assay (Accumetrics, San Diego, CA)





Genotyping:

TaqMan® Drug Metabolism Genotyping Assays

(Life Technologies; Pleasanton, CA)



- loss of function CYP2C19*2 variant (rs4244285)

- other functional variants of CYP2C19 [*3 (rs4986893),*4 (rs28399504),

*5 (rs56337013),*6 (rs72558184), *7 (rs72558186),*8 (rs41291556)

*17 (rs12248560)] and ABCB1 (rs1045642)

CYP2C19 Genotype/Predicted Phenotype Relationships



Genotype Phenotype

*1/*1 WT/WT Extensive

*1/*2-*8 WT/LOF Intermediate

*2-8/*2-*8 LOF/LOF Poor

*17/*17 GOF/GOF Ultra-rapid

*1/*17 WT/GOF Rapid heterozygous

*2-8/*17 LOF/GOF Poor/rapid



Group I- Metabolizer Status

Ultra-rapid Metabolizer (UM) [Ultra-rapid] +[Rapid Het]

Extensive Metabolizer (EM) Extensive

Intermediate Metabolizer (IM) [Intermediate] + [Poor/rapid]

Poor Metabolizer (PM) Poor



Group II- LOF Carrier Status

LOF carriers [Intermediate] + [Poor/rapid ] + [Poor]

LOF non-carriers [Extensive] + [Ultra-rapid] + [Rapid het]



Group III- GOF Carrier Status

GOF Carriers [Ultra-rapid] + [Rapid het]

Extensive Metabolizer (EM) Extensive

LOF carriers [Intermediate] + [Poor/rapid] + [Poor]

CYP2C19 Genotype Frequencies, n (%)

Ticagrelor Clopidogrel Total

(n=92) (n=82) (n=174)

Group 1

Ultra-rapid Metabolizer 27 (30) 28 (24) 55 (32)

Extensive Metabolizer 28 (30) 31 (38) 59 (34)

Intermediate Metabolizer 35 (38) 20 (24)* 55 (32)

Poor Metabolizer 2 (2) 3 (4) 5 (3)

Group II

LOF carrier 37 (40) 23 (28) 60 (35)

LOF non-carrier 55 (60) 59 (72) 114 (66)

Group III

GOF carrier 27 (29) 28 (34) 55 (32)

Extensive Metabolizer 28 (30) 31 (38) 59 (34)

CYP2C19 Diplotype

*1/*1 28 (30) 31 (38) 59 (34)

*1/*2 20 (22) 13 (16) 33 (19)

*1/*3 1 (1) 1 (1) 2 (1)

*1/*17 22 (24) 28 (34) 50 (29)

*2/*2 2 (2) 3 (4) 5 (3)

*2/*17 13 (14) 6 (7) 19 (11)

*8/*17 1 (1) 0 (0) 1 (0)

*17/*17 5 (5) 0 (0)** 5 (3)



* p=0.05, **p=0.04;Ticagrelor vs. Clopidogrel

ABCB1 Genotype/Predicted Phenotype Relationships

and Frequencies, n (%)



Genotype Phenotype

C/C High Expression

Intermediate

C/T

Expression

T/T Low Expression









Ticagrelor Clopidogrel Total

(n=92) (n=82) (n=174)



C/C 30 (33) 21 (26) 51 (29)



C/T 43 (47) 40 (49) 83 (48)



T/T 19 (21) 21 (26) 40 (23)

Predose Platelet Function

5 M ADP-Induced Aggregation

8 hrs After Loading Dose



p-values between genotypes p-values between genotypes p-values between genotypes

80 (UM,EM,IM,PM) (LOF,Non-LOF) (GOF,LOF,EM)

TIG=0.152, CLP=0.289 TIG=0.518, CLP=0.078 TIG=0.117, CLP=0.168

p=0.387

p=0.0006

p<0.0001

p=0.0006

Aggregation (%)









60 p<0.0001 p=0.0006

p<0.0001









40









20









0

UM EM IM PM LOF LOF GOF

Non-carrier



Ticagrelor Clopidogrel

5 M ADP-Induced Aggregation

8 hrs After Last Maintenance Dose



p-values between genotypes p-values between genotypes p-values between genotypes

80 (UM,EM,IM,PM) (LOF,Non-LOF) (GOF,LOF,EM)

TIG=0.95, CLP=0.33 TIG=0.88, CLP=0.097 TIG=0.12, CLP=0.17

p=0.149

Aggregation (%)









p=0.0016 p<0.0001

60 p<0.0001 p<0.0001 p=0.0016



p=0.0004







40









20









0

UM EM IM PM LOF LOF GOF

Non-carrier



Ticagrelor Clopidogrel

20 M ADP-Induced Aggregation

8 hrs After Loading Dose

p-values between genotypes p-values between genotypes p-values between genotypes

80 (UM,EM,IM,PM) (LOF,Non-LOF) (GOF,LOF,EM)

TIG=0.159, CLP=0.306 TIG=0.525, CLP=0.079 TIG=0.209, CLP=0.165

p<0.0001

p=0.0001 p<0.0001

p<0.0001 p=0.149 p=0.0001



p<0.0001

Aggregation (%)









60









40









20









0

UM EM IM PM LOF LOF GOF

Non-carrier



Ticagrelor Clopidogrel

20 M ADP-Induced Aggregation

8 hrs After Last Maintenance Dose



P-values between genotypes P-values between genotypes P-values between genotypes

80 (UM,EM,IM,PM) (LOF,Non-LOF) (GOF,LOF,EM)

TIG=0.864, CLP=0.056 TIG=0.803, CLP 0.049 TIG=0.936, CLP=0.080

p=0.001

p=0.0001 p=0.001

p<0.0001



p=0.139 p<0.0001

60

Aggregation (%)









p=0.0001









40









20









0

UM EM IM PM LOF LOF GOF

Non-carrier



Ticagrelor Clopidogrel

VerifyNow-PRU

8 hrs After Loading Dose

p-values between genotypes p-values between genotypes p-values between genotypes

400 (UM,EM,IM,PM) (LOF,Non-LOF) (GOF,LOF,EM)

TIG=0.552, CLP=0.019 TIG=0.301, CLP=0.010 TIG=0.360, CLP=0.028



p<0.0001 p<0.0001

p<0.0001

p<0.0001

300 p=0.149 p<0.0001

p<0.0001

PRU









200









100









0

UM EM IM PM LOF LOF GOF

Non-carrier



Ticagrelor Clopidogrel

VerifyNow-PRU

8 hrs After Last Maintenance Dose



p-values between genotypes p-values between genotypes p-values between genotypes

400 (UM,EM,IM,PM) (LOF,Non-LOF) (GOF,LOF,EM)

TIG=0.535, CLP=0.006 TIG=0.898, CLP=0.002 TIG=0.905, CLP=0.007

p<0.0001 p<0.0001 p<0.0001



p<0.0001

300 p=0.138

p<0.0001 p<0.0001

PRU









200









100









0

UM EM IM PM LOF LOF GOF

Non-carrier



Ticagrelor Clopidogrel

VASP-PRI:

8 hrs After Loading Dose

p-values between genotypes p-values between genotypes p-values between genotypes

100 (UM,EM,IM,PM) (LOF,Non-LOF) (GOF,LOF,EM)

TIG=0.601, CLP=0.153 TIG=0.616, CLP=0.036 TIG=0.498, CLP=0.109

p<0.0001

p<0.0001 p<0.0001 p<0.0001

p<0.0001

80 p<0.0001

p=0.148

VASP-PRI









60







40







20







0

UM EM IM PM LOF LOF GOF

Non-Carrier





Ticagrelor Clopidogrel

VASP-PRI

8 hrs After Last Maintenance Dose



p-values between genotypes p-values between genotypes p-values between genotypes

100 (UM,EM,IM,PM) (LOF,Non-LOF) (GOF,LOF,EM)

TIG=0.635, CLP=0.069 TIG=0.878, CLP=0.010 TIG=0.828, CLP=0.034

p<0.0001

p<0.0001

p<0.0001

80 p<0.0001

p<0.0001 p<0.0001

p=0.148

VASP-PRI









60







40







20







0

UM EM IM PM LOF LOF GOF

Non-Carrier





Ticagrelor Clopidogrel

Influence of Diplotype on Platelet Function During Maintenance Therapy:

5 M ADP-Induced Aggregation







75

5M ADP-Induced Platelet Aggregation (%)









p=0.004 p<0.0001 p=0.005 p=0.009

60







45







30







15





N/A N/A

0

*1/*1 *1/*2 *1/*3 *1/*17 *2/*2 *2/*17 *8/*17 *17/*17





Ticagrelor Clopidogrel

Influence of Diplotype on Platelet Function During Maintenance Therapy :

20 M ADP-Induced Aggregation



75

20M ADP-Induced Platelet Aggregation (%)









p=0.0001 p<0.0001 p=0.003 p=0.003



60







45







30







15





N/A N/A

0

*1/*1 *1/*2 *1/*3 *1/*17 *2/*2 *2/*17 *8/*17 *17/*17



Within the clopidogrel group, there was an influence of diplotype on platelet function, p=0.09





Ticagrelor Clopidogrel

Influence of Diplotype on Platelet Function During Maintenance Therapy:

VerifyNow P2Y12 Assay



400

p=0.0011

p<0.0001

320 p<0.0001 p<0.0001

P2Y12 Reaction Units









240







160







80





N/A N/A

0

*1/*1 *1/*2 *1/*3 *1/*17 *2/*2 *2/*17 *8/*17 *17/*17



Within the clopidogrel group, there was a significant influence of diplotype on platelet function, p=0.006





Ticagrelor Clopidogrel

Influence of Diplotype on Platelet Function During Maintenance Therapy:

VASP-P Assay



100



p<0.0001

p<0.0001 p<0.0001 p=0.007

80

VASP–PRI (%)









60







40







20





N/A N/A

0

*1/*1 *1/*2 *1/*3 *1/*17 *2/*2 *2/*17 *8/*17 *17/*17



Within the clopidogrel group, there was an influence of diplotype on platelet function, p=0.09



Ticagrelor Clopidogrel

Influence of ABCB1 Genotype on Platelet Function









• No influence of ABCB1 genotype on

platelet reactivity to ADP before treatment or during

treatment in either group by all measurements.

Conclusions

• First report demonstrating the superior antiplatelet effect of

ticagrelor compared to clopidogrel irrespective of CYP2C19 genotype.

• Whereas CYP2C19 genotype influenced the antiplatelet effect of

clopidogrel, there was no effect during ticagrelor therapy.



• Influence of CYP2C19 genotype in clopidogrel-treated patients most

evident during maintenance and by the VerifyNow P2Y12 assay.



• Larger studies needed to examine the relative influences *2 and *17 on

clopidogrel antiplatelet effects.



• Results consistent with:

- overall PLATO trial clinical outcomes

- PLATO genetic substudy:

ticagrelor was more clinically effective than clopidogrel

irrespective of CYP2C19 genotype.

Published online November 15, 2010


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