Comparability of biotheurapeutic products following manufacturing

process improvement

Quality aspects and comparability issues

Inger MollerupCorporate Vice President

Regulatory AffairsNovo Nordisk A/S

Manufacturing of biologics: each process is unique

Transfer Plasmid

into Host Cell

Cell culture orfermentation

Purify DrugSubstance

Formulate Product for

UseDownstream Processing

Clone Gene into DNA Vector

Biosimilar

MaybeA differentformulation

DifferentPurification

protocol

Probably different DNA

vector

Same AA -maybe same

genetic sequence

Different Recombinant

cell system

Different In-process

controls

Slide no 213 Feb 2013

Specifications are linked to the entire manufacturing process (ICH Q6B)

Cell Bank Manuf.process

Drugsubstance

Drugproduct

Q5B Genetic stab.

Q5B Cell substrate

Q5B Viral safety evaluation

Q6B Specifications of biotech product

Q5B Stability of biotech product

Tox studiesClinical studies

Biosimilar Slide no 3

13 Feb 2013

Comparability exercise and Process changes

Change filter supplier

Move equipment different part of facility

Move manufacturing to new facility

New cell line

New Process

New Formulation

Scale-up Manufacturing

Manufacturing Change

•Low risk•Frequent•Supported

by: Analytical and Process Data

•Highest risk•Rare•Extensive Data:

Analytical, Process and Human data

Biosimilars

New:

•DNA?

•Cell line

•Process Technology?

•Fermentation process

•Purification process

•Analytics

•Facilities

•Formulation?

•And - no history

Stepwise Comparability exercise:

Clinical Comparability

Nonclinical Comparability

Chemical Comparability

Character of Change

Complete Comparability exercise:

Clinical Comparability

Nonclinical Comparability

Chemical Comparability

Presentation title Slide no 5Date

How is a biosimilar designed?

Target probably well defined”Design space”?

The Comparability Concept for BiosimilarsAnalytical data + nonclinical and clinical data

Innovator Testing Following a Process Change

Biosimilar Testing

Pre-change Post-change New process

In process controls(15 – 50 tests)

In process controls(15 – 50 tests) No comparison possible

Process validation(5 – 15 tests)

Process validation(5 – 15 tests)

No comparison possible

Stability profile(25 – 50 tests)

Stability profile(25 – 50 tests) No comparison possible

Degradation profile(25 – 50 tests)

Degradation profile(25 – 50 tests)

No comparison possible

Drug substance QC(10 – 25 tests)

Drug substance QC(10 – 25 tests)

No comparison possible

Drug product QC(10 – 25 tests)

Drug product QC(10 – 25 tests)

Drug product QC(10 – 25 tests)

Non-clinical testing(tox and others)

Non-clinical testing(tox and others)

Human studiesEfficacy, safety, immunogenicity

Human studiesEfficacy, safety, immunogenicity

Adapted from: T. Lubiniecki (2005)

Manufacturing Process Steps

Cell expansion

Production

Recovery

Purification

Characterisatio

n

Stability

Drug

Substance

Drug Product

Drug Product

Drug Product

Biosimilars, Overview of Scientific/Regulatory Guidelines and Experience, Inger Mollerup Novo Nordisk Slide no 7Oct 08 2009

Innovator examples

Biomass

Clear broth with secreted insulin precursor

Purified insulin precursor

Insulin ester

Insulin ester

Human Insulin (crude)

Purified Human Insulin

Cell removal

Capture process

Purification

Hydrolysis

Purification

Enzymatic conversion

Novo NordiskExpression system: S. CerevisiaeExpressed molecule: • Insulin precursor (3 AA bridge)No refolding needed

Different •HCP’s•reagents•solvents•pH ranges •etc. etc. etc.

Different Expression

Systems

Eli LillyExpression system: E. ColiExpressed molecule: • Pro-Insulin (35 AA bridge)Unfolding and refolding

Biomass

Inclusion Bodies

Trp-LE'-Met-Proinsulin

Proinsulin (unfolded)

Proinsulin-SSO3

Proinsulin (refolded)

Human Insulin (crude)

Purified Human Insulin

Cell harvesting

Cell disruption

IB recovery

IB dissolution

CNBr cleavage

Oxidative sulfitolysis

Folding, S-S bond formation

Enzymatic conversion

Purification

Biosimilar Slide no 8

13 Feb 2013

HI derivatives originating from fermentation

Asn

Cys

Tyr

Asn

GluLeu

GlnTyrLeuSer

IleCys

Ser

Thr

Cys

Cys

Gln

Glu

Val

Ile

Gly

Pro

Lys

Thr

Tyr

Phe

Phe

Gly

Glu

Arg

Gly

Cys

LeuVal

LeuAla TyrLeu

GluVal

His

Cys

Gly

Ser

Leu

His

Val

Gln

Asn

Phe

S

SS

S S

S

DesPhe(B1)-HI

Thr(A8)-O-mannosyl HI

Thr

DesPhe(B1)-Val(B2)-HI

B1

A10

A21

B30

B20B10

A1

Arg

O-Mannosyl

B0Arg-HI

O-Glycosylation site

Impurities

B29

Gln

Asn

AsnCys

TyrAsn

GluLeuGlnTyrLeuSer

IleCys

Ser

ThrCys

CysGln

GluVal

Ile Gly

Pro

Lys

Thr

TyrPhe

PheGly

GluArg

GlyCys

LeuValLeuAla Tyr

LeuGluVal

His

CysGly

Ser

Leu

His

ValPhe

S

SS

S

LysAla

Ala

S

S

B1

A21

B20B10

A1

B29

Gln

Asn

AsnCys

TyrAsn

GluLeuGlnTyrLeuSer

IleCys

Ser

ThrCys

CysGln

GluVal

Ile Gly

Pro

Lys

Thr

TyrPhe

PheGly

GluArg

GlyCys

LeuValLeuAla Tyr

LeuGluVal

His

CysGly

Ser

Leu

His

ValPhe

S

SS

S

S

S

AlaAlaLysB1

A21

B20B10

A1

B22

Gln

Asn

AsnCys

TyrAsn

GluLeuGlnTyrLeuSer

IleCys

Ser

ThrCys

CysGln

GluVal

Ile Gly

GluArg

GlyCys

LeuValLeuAla Tyr

LeuGluVal

His

CysGly

Ser

Leu

His

ValPhe

S

SS

S

S

S

B1

A21

B20B10

A1B29

Gln

Asn

AsnCys

TyrAsn

GluLeuGlnTyrLeuSer

IleCys

Ser

ThrCys

CysGln

GluVal

Ile Gly

Pro

Lys

ThrTyr

PhePhe

Gly

GluArg

GlyCys

LeuValLeuAla Tyr

LeuGluVal

His

CysGly

Ser

Leu

His

ValPhe

S

SS

S

S

S

B1

A21

B20B10

A1

B29

Gln

Asn

AsnCys

TyrAsn

GluLeuGlnTyrLeuSer

IleCys

Ser

ThrCys

CysGln

GluVal

Ile Gly

Pro

Lys

ThrTyr

PhePhe

Gly

GluArg

GlyCys

LeuValLeuAla Tyr

LeuGluVal

His

CysGly

Ser

Leu

His

ValPhe

S

SS

S

S

S

LysAla

Ala

B1

A21

B20B10

A1

IM1/IM3 IM5

DOI S-comp.

IM2/IM4

Biosimilar

Pharmacopeia compliance is not a guarantee

Test

Specification Method of Analysis

Identification A by Assay Complies Ph Eur, USP

Identification B by Peptide mapping Complies Ph Eur, USP

Bioidentity ≥15 USP U/mg USP

High molecular weight proteins ≤1.0% Ph Eur, USP

Related proteins:Insulin human related substances

A21 desamido insulin human

Insulin human related impurities

≤2.0%≤2.0%

Ph Eur

Zinc (calculated on dried basis)

≤1.0% Atomic absorptionPh Eur

Loss on drying ≤10.0% Ph Eur, USP

Sulphated ash (calculated on dried basis)

≤2.5% Ph Eur

Microbial control, total viable count ≤300 CFU/g Ph Eur, USP

Bacterial endotoxins ≤10 IU/mg Kinetic chromogenic method Ph Eur method D, USP

Assay (calculated on dried basis)

95.0 – 105.0% ≥27.5 USP U/mg

Ph Eur, USP

Slide no 11

13 Feb 2013

Biosimilars, Overview of Scientific/Regulatory Guidelines and Experience, Inger Mollerup Novo Nordisk Slide no 12Oct 08 2009

An Entirely New Manufacturing ProcessNovo Nordisk Case Studies

• Insulin Product• New production cell, DNA, cell bank• Optimised fermentation, recovery and purification processes• New facility

• Quality: • Full CMC + comparability:

Minute differences in impurity profile• Clinical

• Pharmacokinetics/Dynamics• Immunogenicity

• FDA• No safety threshold for new impurities• Not known when “low” is “low enough”

Even for a small product as insulin, clinical safety studies are necessary to demonstrate the absence of clinically meaningful

differences

Biosimilars, Overview of Scientific/Regulatory Guidelines and Experience, Inger Mollerup Novo Nordisk Slide no 13Oct 08 2009

Haemophilia Product (rFVIIa): Innovator Process Change Requiring New BLA

Change in host cell line led to change in glycosylation patternHuman PK study revealed significant difference

• New cell line (CHO) evaluated post approval and compared with original cell line (BHK)

• Quality: Comparability data showed differences in glycosylation

• N-acetyl-galactosamine (GalNAc) present in BHK-rFVIIa only (~20%)

• Slightly lower content of terminal sialic acid residues on BHK-rFVIIa compared with CHO-rFVIIa.

• Clinical• Pharmacokinetic (BE) study• Further data required

• Safety/Efficacy • Immunogenicity

• FDA:• Requires new BLA and substantial

clinical studies

Comparative Human PK*Mean AUC for CHO 30%> than for BHK

* Submitted for publication

Biosimilars, Overview of Scientific/Regulatory Guidelines and Experience, Inger Mollerup Novo Nordisk Slide no 14Oct 08 2009

Myozyme® (rhGAA) Example: Scale Up Resulted in Different Product Characteristics

• Orphan drug produced in CHO cells, both 160 litre and 2000 litre cell culture scale needed for supply

• EMEA:• Concerns with regard to the potential levels of process impurities and

possible immunogenicity of CHO cell impurities for the 160L scale• Impurity profile of 2000L process improved compared to 160L process• Clinical experience from the 2000L process was taken into account• The 2000L product (only) was granted market authorisation

• New facility recently approved• FDA:

• FDA decided that batches manufactured in the two scales are different (incl. differences in glycosylation) and therefore constitute distinct products that must be licensed separately (separate BLA’s) and marketed under different trade names

• Genzyme submitted Lumizyme® for FDA approval

Not yet possible to link quality attributes to clinical S&E

Biosimilars, Overview of Scientific/Regulatory Guidelines and Experience, Inger Mollerup Novo Nordisk Slide no 15Oct 08 2009

Biosimilar Case Studies

Product class-specific Guidelines

EU „Biosimilarity“ GuidelinesEMA Guidelines

Biosimilar

Guideline on similar biological medicinal product

Guidelines on similar biological medical products containing biotechnology derived proteins as active substances

Quality Non-clinical/Clinical

Rh Insulin

Somatropin rG-CSF Epoetin Interferonα LMWH mAbs

2013 reviewAnaloguesHM long acting 2013

review2013 finalise

2013 finalise + β

2013 review

Slide no 16

13 Feb 2013

Bundesinstitut für Arzneimittelund Medizinprodukte

Principles of Biosimilar Approach

„Similarity“ in terms of quality, safety and efficacy

To a reference product licensed in the EU

(Reference product: licensed based on full application, data protection expired)

Same reference product for all aspects of the comparability exercise

Biosimilar Approvals In Europe to Date:Negative opinions and withdrawals

Trade Name Common NameInternational

Nonproprietary Name

Biosimilar Sponsor(s)

Reference Product

Decision Decision Date

Alpheon® Interferon alfa-2a BioPartners

Roferon-A® Negative Opin. Jun 2006

Insulin Rapid Marvel Soluble Insulin Marvel Humulin® Withdrawn Jan 2008

Insulin Long Marvel Isophane Insulin Marvel Humulin® Withdrawn Jan 2008

Insulin 30/70 Mix Marvel

Biphasic Insulin Marvel Humulin® Withdrawn Jan 2008

Epostim® Epoietin Reliance Genemedix

Eprex® Withdrawn Apr 2011

Insulin Solumarv Soluble Insulin Marvel Humulin® Withdrawn Dec 2012

Insulin Isomarv Isophane Insulin Marvel Humulin® Withdrawn Dec 2012

Insulin Combimarv Biphasic Insulin Marvel Humulin® Withdrawn Dec 2012

EMA Public Assessment Report

Marvel Insulins: Human Long, Rapid and Mix (2008)

• Not possible to conclude that purity is comparable to reference product

• Clinical (PK/PD) data: Significant differences in bioavailability compared to reference products

• Products were not biosimilar, file withdrawn from EMA

PD data/Long

PD studies demonstrated clinically meaningful differences Clinical safety and efficacy study confirmed difference

PD data/Mix

* Source: European Public Assessment Report

Biosimilar Slide no 19

13 Feb 2013

Marvel Insulins: Human Long, Rapid and Mix (2012)

• Company reason:• Sufficient time to repeat and

submit bioequivalence T1D PK/PD data on each clamp study

• Comply with the planned new insulin guideline

• Validated CRO• CHMP• Manufacturing concerns• Similarity was questioned

Biosimilar Slide no 20

13 Feb 2013

Biosimilar Approvals In Europe to Date

Trade Name Common NameInternational

Nonproprietary Name

Biosimilar Sponsor(s)

Reference Product

Decision Decision Date

Omnitrope® somatropin Sandoz Genotropin® Approved Apr 2006

Valtropin® somatropin BioPartners Humatrope® Approved Apr 2006

“Sandoz EPO”Abseamed® Epoetin alfa HexalBinocrit®

epoetin alfaMediceHexalSandoz

Eprex® Approved Aug 2007

“Hospira EPO”Silapo® Retacrit®

epoetin zetaStadaHospira

Eprex® Approved Dec 2007

“Teva G-CSF”Tevagrastim® Ratiograstim®

Filgrastim ratiopharm (MA withdrawn July 2011)Biograstim®

FilgrastimTevaRatiopharmCT Arzneimittle

Neupogen® Approved Sep 2008

“Sandoz G-CSF”Filgrastim Hexal Zarzio®

FilgrastimHexalSandoz

Neupogen® Approved Feb 2009

Nivestim® Filgrastim Hospira UK Neupogen® Approved Jun 2010

EMA Public Assessment Report

Biosimilars, Overview of Scientific/Regulatory Guidelines and Experience, Inger Mollerup Novo Nordisk

Slide no 22

Oct 08 2009

EU: Early version of Omnitrope® (somatropin)Reference product: Genotropin (Pfizer)

• Early version of the product: 57% of patients developed antibodies against Omnitrope• Problem was residual host-cell protein

• Re-developed purification process• Conducted a second phase 3 study• Antibody levels reduced (comparable)• Approved by EMEA, FDA, TGA – and recently by Health Canada

and PMDA in Japan

Link between quality parameter (HCP) and clinical safety (immunogenicity) found in clinical studies

* Source: European Public Assessment Report

Biosimilars, Overview of Scientific/Regulatory Guidelines and Experience, Inger Mollerup Novo Nordisk

Slide no 23

Oct 08 2009

EU: Alpheon (alpha interferon)Reference product: Roferon (Roche)

• Differences in impurity profile were observed

• Key Clinical Data• PK (3 studies):

supra-bioavilability(early study); comparable; inconclusive• PD (2 studies): PD equivalence/no PD equivalence• Safety&Efficacy:

• Clinically and statistically significant difference in virological relapse rate found: more patients on Alpheon had relapse

• Different rate of adverse events and laboratory-related events judged as clinically relevant

Clinical studies evaluating efficacy and safety demonstrated clinically meaningful differences leading to Rejection by EMEA

* Source: European Public Assessment Report

Biosimilars, Overview of Scientific/Regulatory Guidelines and Experience, Inger Mollerup Novo Nordisk Slide no 24Oct 08 2009

Other products - not approved under biosimilar framework

Biosimilars, Overview of Scientific/Regulatory Guidelines and Experience, Inger Mollerup Novo Nordisk Slide no 25Oct 08 2009

”Ex-EU” Somatropin: New Thioether Variant Identified

• Thioether variant found in some hGH products (up to ~30%)

• Hormotrop®, Yelit®, Cryotropin®• Thioether variant not identified in

• Saizen®• International standards:

NIBSC 98/574 (r), NIBSC 80/505 (p), EP r-hGH CRS

• Thioether variant: • Not detected by compendial and other

existing chromatographic methods – new methods required

• Generated by high pH at elevated temparature (40 deg C)• Significantly reduced biopotency in rat model

• Analytical methods must be ”tailor made”

* Lispi, M. et al, J. Pharm. Sciences, DOI 10 1002/jps 21774, 2009

New impurity identified with reduced biopotency Change in Clinical efficacy & safety cannot be excluded

Biosimilars, Overview of Scientific/Regulatory Guidelines and Experience, Inger Mollerup Novo Nordisk Slide no 26Oct 08 2009

Conclusion: Quality Aspects and Comparability

• Products must have highly similar molecular structural features

• Amino acid sequence must be the same• Glycosylation patterns must be highly similar

• Same host cell type system

• Impurity profiles must be highly similar• Analytical methods extremely important

• Only way to exclude clinically meaningful differences in efficacy, safety and immunogenic potential – is from clinical data

• Multiple examples illustrate close link between chemical and clinical comparability

• Can’t be predicted from structure

• Full comparability exercise needed – quality, nonclinical and clinical – as outlined in EMEA guidance documents