Critical aspects regarding the application of GLP ... · Critical aspects regarding the application...

OECD OECD EventEvent , Villa , Villa TuscolanaTuscolana , Frascati (Roma), , Frascati (Roma), ItalyItaly , , AprilApril 10 10 –– 11,11, 2008 2008

Critical aspects regarding the Critical aspects regarding the application of GLP Principles to application of GLP Principles to

new compounds such as new compounds such as biotechnology products biotechnology products

MariaMaria Mercede BrunettiMercede Brunetti

RTC RTC SpASpA ,, PomeziaPomezia , Rome, Italy, Rome, Italyee--mail:mail: mercebrmercebr @@rtcrtc .it.it

CoordinatorCoordinator --GLP group of GIQARGLP group of GIQAR


INTRODUCTION

• Introduction

• Applicable GLP Principles & guidelines

• Conclusions - Final considerations

• Examples (cases)


INTRODUCTION

Biopharmaceutical vs conventional products

• Biological & structural properties of product

� macromolecules (proteins, complexbiopolymers) vs small molecules (organic)

� biological/biotechnology-derived vsorganic synthesis


• Study types: in vitro test /analytical methods

� Test & reference items� Analytical aspects

� characterisation� analysis of doses� analysis of biological matrixes

(TK/PK, immunogenicity)

Biopharmaceutical vs conventional products

INTRODUCTION


GLP (OECD, 1997)

Applicable GLP Principles

GLP (Italy, 2007)


OECD Advisory Document No.: 14, 2004

Applicable GUIDELINES (1)



ICH 6S, 1998

Farmacogenomics, 2005

Immunogenicity, 2008


FDA - Bioanalytical method validation, 2001

Workshop - AAPS/FDA, 2007

Immunoassay, 2004

FDA Warning letters



OECD Advisory Document No.: 14, 2004


PURPOSE: To facilitate the proper application and interpretation of the GLP Principles for the organisation and management of in vitro studies

• standard regulatory tests

• new methods/areas ( ...omics)

• high trough-put screening tests

OECD Advisory Document No.: 14


APPLICATION

• Standard in vitro studies



• in vitro methods (part of in vivo studies)APPLICATION



EXAMPLES(cases)


CASE - 1a

•Test item protein (vaccine, Ab)

•Type of study 13 wks in...

•Study Plan includes the evaluation of immunogenicity(in responsive specie)


CASE - 1b

• REFERENCE ITEM ?

• ELISA TEST ?


CASE-1c: reference item

Ab: Antibody/reference item

• Preparation• Purification• Quantitation and ...


Ab: Preparation

• Must be prepared in a GLP study ?

CASE-1d: reference item


Ab : Purification

GLP - characterisation & labelling:

• identity , concentration, storage condition , preparation date, expiry/retest date

CASE - 1e: reference item


Ab: Quantitation and...

Ex.: in mg of protein/ml -Lowry

CASE-1f: reference item



Characterisation: meaning, extension ?


FDA Guidance - Bioanalytical Method Validation (2001) - in house STD:• source and lot number, expiration date, certificate of analysis when available, and/or evidence of identity and purity

Characterisation: meaning, extension ?



CASE-1h: reference item


CASE-1i: ELISA TEST

• set up• validation• analysis


CASE-1l: ELISA TEST

SET UP ad hoc per studio


CASE-1m: ELISA TEST

VALIDATION• Precision• Accuracy• LOQ (L/U) & range• Calibration curve• Matrix & buffer effects• Stability (freezing)

• acceptancecriteria

• reagents in house

• stability


ANALYSES

• Storage ofsamples

CASE-1n: ELISA TEST

• acceptancecriteria

• reagents in house

• stability


CASE-2: control matrix

� Characterisation ?

� Stability/retest date ?

� endogenous product ?


CASE - 3a

• Test item: recombinant protein (therapeutic use )

• Type of study: repeated toxicity

• Study Plan includes:� evaluation of immunogenicity

(immuno-response present/absent )

� kinetic analysis (non-chromatographic methods)


CASE - 3b

2 TYPES of TEST

• set up• validation• analysis

A: immunogenicity(test item + reference item - biosimilar)

B: kinetic (test item + reference item - biosimilar)


CASE - 3c

☯ ELISA test in house ☯ Commercial Ab - surrogate

(mono or policlonal ?)

(antigen : test item in vivo study and/or reference item -biosimilar)

☯ Response: YES/NO (cut point definition)

A: immunogenicity - qualitative/semiquantitative(test item + reference item - biosimilar)


CASE - 4a

• Test item DNA vaccine

• Type of studyrepeated toxicity

• Study Plan includes: biodistribution and evaluation ofimmunogenicity(in responsive strain)


CASE - 4b

A: biodistribution: PCR(Sponsor - non GLP site in a multisite study-OK phase I)

B: immunogenicity humoral and cell-mediated (validation ? acceptance criteria ? scientific publications ?)

2 TYPE of TEST


CASE - 4c

Semiquantitative analysis

☯ Humoral immunogenicity test� Reference item: commercial monoclonal Ab -

surrogate (calibration curve , QC)

� “ Test item ” : OGM (Balb 3T3/...antigen) - supplied by the Sponsor (through University)

Verification at receipt - acceptance (FACS- surrogate)

� Results analyses: criteria ?


CASE - 4d

☯ cell-mediated immunogenicity test� Primary cells (splenocytes) conjugated or not with

peptide + fluorochrome (antigen - high and low concentration)

� Inoculum in treated mice: fluorescence measurement in samples (FACS)

� Results analyses: criteria ?

GLP: enough internal preliminary test/s ?

Semiquantitative analysis


CASE - 5

� qualitative, quantitative andsemiquantitative analyses(PCR, FACS, ELISA, Multiplex, …)

Type of test: Biomarkers (exploratory)


• SCIENTIFIC PERSONNEL (SD)

• PERSONNEL - Technicians

Human Factors: manual activities, variability

FINAL CONSIDERATION


FINAL CONSIDERATION


FINAL CONSIDERATION

RELATE WITH AUTHORITIESRELATE WITH AUTHORITIESRELATE WITH AUTHORITIESRELATE WITH AUTHORITIES


THANKS to :

S. Cinelli & I. Andreini

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