at the crossroads of comparability and characterization...

Pfizer BioTherapeutics Pharmaceutical Sciences

Laura Bass, Katherine Arch-Douglas, Michele Bailey Piatchek, Stephanie Ferrari, Parag

Kolhe, Sue Leander, Khurram Sunasara, Roberto Rodriguez, Lavinia Lewis

Pfizer BioTherapeutics Pharmaceutical Sciences

Analytical Research and Development

St. Louis, MO

At the crossroads of comparability and characterization:

The case study for Trumenba® –

A vaccine for invasive meningococcal disease

WCBP 2015

Outline

• Introducing Trumenba®

• Process changes triggering the comparability study

• Characterization studies performed to enhance product/process

understanding

• Components of the comparability strategy

• Lot selection

• Testing Strategy

• Approach to assessing comparability

• Lessons Learned

Label Category Target

INDICATIONS AND

Vaccine indicated for active immunization to prevent invasive

meningococcal disease caused by Neisseria meningitidis

serogroup B expressing either subfamily A or subfamily B fHBP

in persons 10-25 yrs of age

DISEASE PROFILE

AND INCIDENCE

Meningococcal disease can progress from initial symptoms to

death within 24 hours. Symptoms are similar to the common cold.

Despite the availability of antibiotic treatment, between 10-15% of

patients with meningococcal disease die and 11-19% of those

who survive are afflicted with long-term disabilities, such as

brain damage, hearing loss, learning disabilities or limb

amputations

Meningitis due to Neisseria meningitidis serogroup B accounts for

~ 40 percent of all cases of meningococcal disease

DOSAGE FORMS 0.5 ml of 60 µg of each Subfamily A and Subfamily B lipoprotein,

Histidine buffered saline, Polysorbate 80, aluminum phosphate,

pH=6, in a pre-filled syringe

DOSAGE REGIMEN 3-dose schedule of 0,2,6 months.

Trumenba®; Bivalent rLP2086 Vaccine

Mechanism of Action • fHBP/LP2086 binds human factor H &

protects bacteria from complement attack

• Anti-LP2086 antibodies generated by

vaccine protect through inhibition of factor H binding allowing for complement bactericidal action

Drug Substance • Two rLP2086 lipoproteins (~28 kDa)

manufactured as separate drug substance

• Formulation includes PS80 which likely results in micellar formation and an apparent MW of ~600-700 kDa

Madico et al., 2006; Schneider et al., 2006;

Mascioni et al., 2009; Seib et al., 2009;

Ala’Aldeen et al., 2010; McNeil et al., 2009;

Jacobsson, Mölling & Olcen, 2009

Drug Product • Manufactured by mixing and diluting the

two rLP2086 proteins (subfamily A & B)

with stabilizer (aluminum phosphate)

Drug Substance and Drug Product Process Changes

Change Phase 1/2 Phase 2/3

DS: 120 L

DP: 4L, 10 L or 19 L

(glass vessels)

DS: 2400 L

DP: 150 L

(stainless steel tanks)

Manufacturing

DS: 2 Pilot scale

DP: 2 Pilot Scale

DS: 1 Commercial scale

DP: 2 Commercial scale

DS Storage

250 mL polypropylene (PP) bottle stored

at –70 ± 10°C

5-liter ethylene vinyl acetate

(EVA) bag stored at -55 ± 8°C

Formulation

Succinate Histidine

NaCl NaCl

PS80 PS80 (decreased conc)

AlPO4 AlPO4 (increased conc)

pH 6.0 pH 6.0

Histidine

pH 6.0

DP container

Vial Pre-filled syringe

Pre-filled syringe

DS Comparability Strategy Overview

Assess risk to product quality/consistency/potency

Lipid isoforms were identified early in development as a source of molecular

heterogeneity

Literature data indicated O-acyl-linked lipids were critical to potency

Product and Process Characterization • Leverage heightened product

characterization data to assess risk

associated with process changes

• Example #1: structure/function

characterization of lipids

• Understand impact of process changes on

quality attributes

• Example #2: bioreactor conditions

and purity / lipid isoform profile

Comparability Study • Testing Strategy

• Lot selection

• Strategy for evaluation of

comparability

DS Characterization Invest early in product understanding Example#1; Heightened characterization of lipoproteins

Assess function

of proteins In Vitro Relative Antigenicity

In Vivo Potency

Native &

Degraded

Assess function

of lipids

In Vivo Potency

TLR2 Receptor Assay

Native &

De-O-Acylated

Confirm Primary

Structure of

lipoprotein and

Identify the major

and minor product

isoforms

Peptide level: Non-reduced peptide

mapping of Lys-C proteolytic fragments

by RP-UHPLC/UHR-ESI-Orbitrap MS

Intact level: Molecular mass

determination for intact molecule by

RP-UHPLC/UHR-ESI-QTOF MS

Confirm structure

of lipids

GC/MS of released derivatized fatty acids

LC-MS of lipase digested lipoprotein

DS Process Characterization; Invest Early in Process Understanding Example #2: Impact of bioreactor conditions on Lipid Isoform Profile

Challenges:

Very tight lipid profile observed for clinical lots

manufactured early in development

Bioreactor conditions modified during scale up

Resolution:

Define operating space for lipid isoform profile

Execute multifactor fermentation DOE using qualified scale down

models and monitor lipid profiles as an output.

Define process parameter ranges to consistently achieve desired

lipid peak ratio as monitored by RP-HPLC

Early process development data indicated potential link between

bioreactor conditions and lipid isoform profile

Consistency of lipid isoform profile is critical to potency

Lipid Profile RP-HPLC for lipid isoform profile

3 Major, 5 Minor

overlay_TS

Project Name: H210_SHARE_PHARMSCIReported by User: Tanya Shang (TShang)

Report Method: overlay_TS Printed 4:17:42 PM 2/4/2011

Sample Name: 17Nov10_A_A1002; Date Acquired: 11/17/2010 5:29:00 PM; Injection Volume 10.00; Acq Method Set rLP2086_RP_STM7187_MSSample Name: 17Nov10_A_A1003; Date Acquired: 11/17/2010 6:30:00 PM; Injection Volume 10.00; Acq Method Set rLP2086_RP_STM7187_MSSample Name: 17Nov10_A_B302; Date Acquired: 11/17/2010 7:31:00 PM; Injection Volume 10.00; Acq Method Set rLP2086_RP_STM7187_MSSample Name: 17Nov10_A_B303; Date Acquired: 11/17/2010 8:32:10 PM; Injection Volume 17.00; Acq Method Set rLP2086_RP_STM7187_MSSample Name: 17Nov10_A_B304; Date Acquired: 11/17/2010 9:33:10 PM; Injection Volume 10.00; Acq Method Set rLP2086_RP_STM7187_MS

Minutes

5.00 10.00 15.00 20.00 25.00 30.00 35.00

4MnB2086A-1002

Degradants De-O-Acylated

AB C D E

4MnB2086A-1003

G10700B302

G10700B303

G10700B304

overlay_TS

Minutes

5.00 10.00 15.00 20.00 25.00 30.00 35.00

4MnB2086A-1002

AB C D E

4MnB2086A-1003

G10700B302

G10700B303

G10700B304

Site 2 pilot

Pilot Scale batch#1

Pilot Scale batch#2

Commercial Scale batch#1

DS Process Characterization Lipid Peak Ratio: Peak 3 [C16:0, C18:1, C16:0]

• Manufacturing batches run at center points

• Fermentation DOE data were used to define operating ranges at

manufacturing scale to deliver consistent lipid profiles

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17

rmalized

Batch Number

Lipid Peak 3 Process Control

peak 3

Pk3 Prcs Cap: high

Pk3 Prcs Cap: low

DS Comparability Lot Selection

• Lot selection included planned use of the DS materials

• Key CMC milestones; PV, Stability, Reference Std

• Key Clinical milestones: Ph1, Ph2, Ph3

Batch Scale Manufacture Site Batch Use

10 L Site#1 Phase 1, Non-clinical Toxicology

120 L Site#1 Phase 2, Non-clinical Toxicology

120 L (2 batches) Site#1 Phase 2 Clinical

2400 L (6 batches) Site#2/Commercial Demonstration

2400 L Site#2/Commercial Process Validation

2400 L Site#2/Commercial Process Validation, Primary Stability

2400 L Site#2/Commercial Process Validation, Primary Stability,

Reference Material

2400 L Site#2/Commercial Process Validation, Primary Stability

2400 L

(2 batches)

Site#2/Commercial Phase 2 and Phase 3 Clinical, Stability

2400 L Site#2/Commercial Phase 3 Clinical, Stability

2400 L (3 batches) Site#2/Commercial Stability

DS Comparability Testing Strategy

Type of Testing Data Provided

Release

Tabulated data for CQA’s in 3.2.S.2.6

Tabulated data for all attributes in

3.2.S.4.4

Stability

Tabulated data for all attributes in

3.2.S.7.3

Characterization

Tabulated data and overlays of profiles

provided in 3.2.S.2.6

DS Comparability Testing Strategy; Characterization Testing

Test Executed Information Provided

Intact Molecular Mass by

RP-HPLC/UHR-ESI-QTOF MS

Confirm primary structure and post-

translational modifications at the

intact protein level

Non-reduced peptide mapping by

RP-HPLC/UHR-ESI-QTOF MS

Confirm primary structure and identify

post-translational modifications at the

peptide level

Lipidated N-terminal peptide

profiling by organic SE-HPLC/UHR-

ESI-QTOF MS

Identify structures of N-terminal

lipopeptide isoforms

GC/MS of released fatty acids

Confirm fatty acid composition

Far-UV CD

Assess secondary structure

Near-UV CD

Assess tertiary structure

DS Comparability Assessment Characterization Testing; Peptide Map Profiles

overlay_TS

Sample Name: 15Nov10_A_A1002; Date Acquired: 11/16/2010 3:03:48 PM; Acq Method Set rLP2086_map_Hclass

Sample Name: 15Nov10_A_A1003; Date Acquired: 11/16/2010 10:53:16 AM; Acq Method Set rLP2086_map_Hclass

Sample Name: 15Nov10_A_B302; Date Acquired: 11/16/2010 11:55:38 AM; Acq Method Set rLP2086_map_Hclass

Sample Name: 15Nov10_A_B303; Date Acquired: 11/16/2010 4:06:28 PM; Acq Method Set rLP2086_map_Hclass

Minutes

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 45.00

4MnB2086A-1002

4MnB2086A-1003

G10700B302

G10700B303

G10700B304

overlay_TS

Sample Name: 15Nov10_A_A1002; Date Acquired: 11/16/2010 3:03:48 PM; Acq Method Set rLP2086_map_Hclass

Sample Name: 15Nov10_A_A1003; Date Acquired: 11/16/2010 10:53:16 AM; Acq Method Set rLP2086_map_Hclass

Sample Name: 15Nov10_A_B302; Date Acquired: 11/16/2010 11:55:38 AM; Acq Method Set rLP2086_map_Hclass

Minutes

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 45.00

4MnB2086A-1002

4MnB2086A-1003

G10700B302

G10700B303

G10700B304

DS Comparability Assessment Release Testing; RP-HPLC for purity, lipid isoform profile and degradants

overlay_TS

Minutes

5.00 10.00 15.00 20.00 25.00 30.00 35.00

4MnB2086A-1002

AB C D E

4MnB2086A-1003

G10700B302

G10700B303

G10700B304

overlay_TS

Minutes

5.00 10.00 15.00 20.00 25.00 30.00 35.00

4MnB2086A-1002

AB C D E

4MnB2086A-1003

G10700B302

G10700B303

G10700B304

Site 2 pilot

Pilot Scale batch#2

Pilot Scale batch#1

DS Comparability Assessment Conclusions

Type of Testing Assessment of Comparability

Release

• All results met specification acceptance criteria

and no data trends observed across lots tested.

• Results were comparable across batches.

Stability

• All results met specification acceptance criteria

and no data trends observed across lots tested.

• Results were comparable across batches.

Characterization

• Qualitative assessment confirms comparability of

profiles and tabulated data for each individual

attribute tested.

DP Process Changes

Change Phase 1/2 Phase 2/3

DP: 4L, 10 L or 19 L

(glass vessels)

DP: 150 L

(stainless steel tanks)

Manufacturing

DP: Site#1, Site#2

Formulation

Succinate Histidine

NaCl NaCl

PS80 PS80 (decreased conc)

AlPO4 AlPO4 (increased conc)

pH 6.0 pH 6.0

Histidine

pH 6.0

DP container

Vial Pre-filled syringe

Pre-filled syringe

DP Comparability Process Characterization Well-controlled and characterized process delivers consistent product

“Building in” Comparability During Site Change

• DeRisk impact of changes via “at scale” process characterization studies and strategic process fitting

• Execute process at commercial site and scale at high & low end of NOR (process charact.)

• “Fit” process into new site using same scale and equipment allowing for identical target ranges

Low end

High end

Target

Normal Operating Range (NOR)

0 0.5 1 1.5 2 2.5 3 3.5

Sub A- Low Range

Sub B- Low Range

Sub A- High Range

Sub B- High Range

0 0.5 1 1.5 2 2.5 3 3.5

Sub A- Low Range

Sub B- Low Range

Sub A- High Range

Sub B- HIgh Range

Beginning Middle End Beginning Middle End

Low Spec

High Spec

DP Comparability Lot selection and Testing Strategy

• Lot Selection • Scale: Pilot and Commercial

• Site: Site#2 Pilot, Site#3 Commercial, Site#4 Commercial

• Use: Ph2, Ph3 (lot consistency), PV, Primary Stability

• Testing Strategy:

Type of

Testing

Lots tested Data Provided

Release

All release/stability

lots included in

comparability

assessment

n=5 Site#2

n=7 Site#3

n=9 Site#4

Link to 3.2.P.5.4 Batch analysis

Tabulated data for CQA’s in

3.2.P.2.3

Graphs of CQA results for

holistic data comparison in

3.2.P.2.3

Stability

Link to 3.2.P.8.3 Stability

DP Comparability DP Release Data

Pfizer, Pearl River Vetter, Germany Pfizer, Ireland

Site#2 Pilot Site#1

Commercial Site#2

Commercial

Site#1

Commercial

Site#2

Commercial Site#2 Pilot

DP Comparability Data DP Stability Data

Site#1

Commercial Site#2

Commercial

DP Comparability Assessment Conclusions

Type of Testing Assessment of Comparability

Release

• All results met commercial acceptance criteria and

no data trends observed across lots tested

• Results were comparable across batches

Stability

• All results met commercial acceptance criteria and

no data trends observed across lots tested

• Results were comparable across batches

Comparability Strategy Lessons Learned

Invest early in process characterization Highlight understanding of link between process parameters and key quality

attributes

Invest early in heightened product characterization Provide clear summary of product understanding with a focus on CQA’s

Go beyond tabulation of data

Leverage process/product characterization data to design comparability

strategy

Be Strategic in lot selection

Comparability Strategy Lessons Learned

• Have Frequent and open dialogue with the regulatory authorities!

Robust CMC package supported 18 month acceleration of BLA after FDA

granted Breakthrough Therapy Designation

BLA Planning started ~3 yrs prior to targeted filing date

BLA Authoring started ~2 yrs prior to targeted filing date

CMC sections of BLA submitted 5 mos after BTD granted

FDA feedback on initial comparability package:

Does the Agency agree that Pfizer has demonstrated the

comparability of pilot scale materials and commercial scale

materials for drug substance and drug product?

“No, there is insufficient commercial scale lot information” **

FDA feedback on comparability package submitted in BLA briefing pkg:

CBER agrees that, pending the potency results and a thorough review, the

comparability data among vaccine lots prepared using the commercial

manufacturing scale may support an “Accelerated Approval” submission

Acknowledgements

• Herb Runnels

• Dave Cooper

• Arthur Duffy,

• Zhaowei Jin

• Willy Tan

• Rob Dufield

• Tanya Shang

• John Whittemore

• Marja van Zeijl

• Elisabeth Crandall

• Claire Roche

• Marta Czupryn

• Sriram Srinivasan

• Jean Crinean

• John Cundy

• Kathrin Jansen

• Emilio Emini

Abstract

• Although the frequently cited paradigm “the process is the product” is often applied rigorously

for vaccines, application of an extensive analytical toolkit is required to gain an in-depth

understanding of the process and the product (and ultimately to demonstrate

comparability). During vaccine development the goal is to minimize process changes to

ensure delivery of a product with a consistent quality and safety profile. The driver for

minimizing process changes is often due to the limitations of the analytical tools available and

difficulty associated with assessing the impact of process changes on vaccine product quality

attributes and associated clinical outcomes (an appropriate clinical surrogate is often not

available). While the goal for vaccine development is to lock the process early, changes are

often inevitable and therefore comparability studies are unavoidable. If characterization

investments are not ramped up until the later stages of development, you may encounter a

crossroad where the analytical toolkit used for process/product understanding is inadequate

for establishing comparability.

• This presentation provides, as a case study, the comparability strategy executed for a

bivalent meningococcal serogroup B (MnB) vaccine. The MnB vaccine development

example will illustrate how, consistent with quality by design principles, the application of an

extensive analytical toolkit facilitated the in-depth characterization of both the process and

product. In addition, this case study will highlight how delivery of a robust comparability

package enabled the project to advance to Phase 3 and ultimately enabled the accelerated

approval of this vaccine.

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