Feasibility, validation, implementation: 3 pillars for ... · Intra-assay precision 4% CV...

Copyright © 2015 Actelion Pharmaceuticals Ltd

Feasibility, validation, implementation:

3 pillars for quality biomarker results in

clinical studies

Hervé Farine

Department of Translational Science

© 2015 Actelion Pharmaceuticals Ltd

Prostaglandin D2 receptor (CRTH2)

• Chemoattractant-Receptor homologous molecule expressed on Th2 cells

(CRTH2)

• present in blood cells on:

• agonist prostaglandin D2 (PGD2)

• leads to cell migration and activation (e.g.: release of cytokines)

Eosinophils Basophils Th2 cells

Source: Blausen.com staff

01 Dec 2015 1


Receptor internalization assay

CRTH2 antagonist

(ACT-453859) agonist

(dk-PGD2)

Whole blood

basophils

staining antibody

Stimulation Staining Flow cytometric

measurement

01 Dec 2015 2


Receptor internalization assay

c o n c e n tra t io n (n M )

CR

TH

2 (

MF

I)

1 0 -1 1 0 0 1 0 1 1 0 2 1 0 3 1 0 4 1 0 5

0

5 0 0

1 0 0 0

1 5 0 0 E o s in o p h ils

B a s o p h ils

CC

R3

SSC

CRTH2

co

un

ts

Concentration-dependent internalization of CRTH2 on basophils and eosinophils.

Strasser et al. NHTM 2015

01 Dec 2015 3


FEASIBILITY

01 Dec 2015 4


Feasibility study

Study that uses samples which represent the expected clinical situation

Decision point 1:

Is the assay capable to measure the analyte/effect?

Is the assay usable in practice?

Should we validate the assay?

01 Dec 2015 5


Feasibility study setup

Spiking with

100 nM antagonist

+

Sample preparation

Volunteer donation

from 09.00-16.00

at Red Cross

Flow cytometry

measurement

4°C

hourly pick-up

by Actelion employee

anonymization

Receptor

internalization

assay

01 Dec 2015 6


Feasibility study in 53 healthy donors

Assay is capable to measure the PD effect and usable for the expected sample throughput.

normalization

01 Dec 2015 7


VALIDATION

01 Dec 2015 8


Validation

Validation is the proof via extensive laboratory investigations that the

performance characteristics of an analytical test system are suitable and reliable for

its intended use.

Decision point 2:

Is the assay reliable for the intended applications?

01 Dec 2015 9


Validation overview

Validation parameters Results

Cell selection with CCR3 102% recovery(CD203c)

Specificity of CRTH2 antibody (Clone BM16) FMO < isotype control <

minimum control

Intra-assay precision 4% CV

Inter-assay precision 12% CV

Post-collection sample stability 24 h

Stability of stained samples 48 h

Post-collection and stained samples stability 2 h + 24 h

01 Dec 2015 10


IMPLEMENTATION

01 Dec 2015 11


Implementation and the associated challenges

Different laboratory

cultures / practices

Different languages /

countries Different

levels of QA Distance /

travel / cost

Different

interpretation

of guidance

Personnel

changes

Inspired by Galen Radebaugh 2010

01 Dec 2015 12


Method transfer process

yes

no

CRO selection

Analytical procedure review & inspection of the facilities

Method familiarization

Feasibility?

Transfer report

Clinical trial

yes

yes

Satisfying

results?

Lab fully

capable?

no

no

01 Dec 2015 13


Implementation in phase I clinical trial

Sample preparation Sample shipment Sample measurement

01 Dec 2015 14


RESULTS

01 Dec 2015 15


Single ascending dose study results

A dose dependent PD effect was observed with a maximum effect reached 2h after dosing

% C

RT

H2 b

loc

ka

de

Gehin et al. J Clin Parmacol 2015

01 Dec 2015 16


PD effect after 24 hours

The %CV of each group were between 5 and 22%

01 Dec 2015 17


Summary

3 pillars for quality biomarker results in clinical studies

FEASIBILITY

We are able to

measure the

required effect /

analyte

VALIDATION

We do the correct assay.

The CRO is capable to

perform correctly the

assay.

IMPLEMENTATION

01 Dec 2015 18

© 2015 Actelion Pharmaceuticals Ltd 01 Dec 2015 19

Acknowledgement

Drug Discovery

Daniel Strasser

Martin Holdener

Peter Groenen

René Roscher

Clinical Development

Jasper Dingemanse

Jochen Zisowsky

Martine Gehin

Patricia Sidharta

Sandrine Gioria

Feasibility, validation, implementation: 3 pillars for ... · Intra-assay precision 4% CV...

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