Operation of a pilot plant for clinical lots of ...Operation of a pilot plant for clinical lots of biopharmaceuticals Client types: • Academic institutions (1990’s) – starting

Operation of a pilot plant for clinical lots of biopharmaceuticals

Neophytos Papamichael, PhD

Quality Assurance GMP | Braunschweig

Workshop “Biopharmaceutical process development and regulatory issues”

Universidade Federal do Rio de Janeiro,

6th.-7th. August 2009


Contents

• Introduction to the Fraunhofer ITEM and Division

• GMP pilot plants

• Quality system requirements in a development environment

• Project development overview and examples

• Outlook


Introduction


Fraunhofer Institute for Toxicology and Experimental MedicineHannover, Germany, www.item.fraunhofer.de

The institute in figures (2008):

Founded 1981

Employees 246

Institute budget € 22 million

(Industrial funding 50%)

Laboratory/office space approx. 7,500 m²

Fraunhofer Institute for Toxicology and Experimental Medicine | Pharmaceutical Biotechnology Division, Braunschweig SiteBraunschweig, Germany, www.item.fraunhofer.de

The division in figures (2008):

Transfer to ITEM 2008

Employees 40

Budget 2008-2013 € 27 million

(Payroll/Investment/Building)

GMP plants 3


Campus of


AachenSchmallenberg

Hannover

St. Ingbert

Stuttgart

Leipzig

Golm

Freising

Fraunhofer Life Sciences Group

IBMT (Biomedical Engineering)IGB (Interfacial Engineering and Biotechnology)IME (Molecular Biology and Applied Ecology)ITEM (Toxicology and Experimental Medicine)IZI (Cell Therapy and Immunology)IVV (Process Engineering and Packaging)

ITEM embedded in and chairs the Fraunhofer Life Sciences Group


•Screening-assay for antimycoticsubstances

•Databases (gene expression profiles)

•DNA-arrays (breast cancer, pathogenic fungi)

•2-D-DNA-gel-electrophoresis for universal differential transcription analysis

•Cartridge for protein and nucleic acid isolation using magnetic beads

•Vascular model demonstrating blood vessel flow characteristics •Demonstration of 3 separate brain models for stroke cell therapy

•Pulsatile bioreactor for physiological in-vitro-cultivation of 3D-tissues

•3D-skin model

•Cell therapy

•Clinical trials phases I and II: asthma, allergic rhinitis, COPD

•Segmental lung application

•Development of API manufacturing processes

•GMP manufacturing of APIs and drugs for clinical trials

ITEM embedded in and chairs the Fraunhofer Life Sciences Group:


•Founded 1966as GMBF•Natural products•Strong engineeringBiochemical Engineering Division

•1995: Shift towards molecular biology•Establishment of GMP unit

•2001: Shift focus to infection research

•2006: Name change to reflect research focus

•1.1.2008: Transfer to

National research centre for biochemical engineering

Pharmaceutical Biotechnology Division


Client types:

• Academic institutions (1990’s) – starting development of biopharmaceuticals (very early entry in product life cycle, poor understanding of costs involved and effort required)

• Small biotech startups (1990’s) – needed access to scale-up and clean room facilities

• Biotech bubble bursts 2001! Financing became difficult.

• Generic manufacturers (2000-2006) - seeking market potential in off-patent biologicals (biogenerics, biosimilars)

• Virtual biotech companies (2004-now) – require complete product development after drug discovery (small scale, pilot scale, transfer to GMP production)


Pharmaceutical Biotechnology Division – bridging the gap in the FhG ITEM GXP Platform...

Processdevelopment /

GMPmanufacturing

GCPclinical trials phases I/II

GLPpre-clinical development

GXP


Pharmaceutical Biotechnology Division - Scope of operation:

• GMP-Cell banking (MCB/WCB generation and storage in LN2 atmosphere)

• Culture medium development and optimisation• API manufacturing process development (USP, DSP)• Process validation studies• Cleaning validation studies• Analytical procedure development and validation to ICH guidelines• Stability studies for purified bulk and intermediates• Non-GMP (pre-clinical) and GMP manufacturing of APIs• GMP-manufacturing of final dosage forms for clinical trials

(individual therapy; future expansion to ampoules, vials and small bottles to 50 ml planned)

• Consultancy services including mock auditing

• Pharmaceutical documentation (development reports, validation reports, master batch production and control records, …)


GMP pilot plants


GMP-Pilot Plants .• GMP I: 460 m², operative 1997 (microbial fermentation, animal cell culture,

purification)

• GMP II: 930 m², operative 2008 (microbial fermentation, animal cell culture, purification, cell banking). Planned extension of GMP II to include aseptic Fill & Finish (vials and ampoules; capacity 60 min-1; online 2010/2011)

• GMP III: 130 m², operative 2001 (cell banking)

• Aseptic Fill & Finish (Hannover) : 100 m² (small scale supply of clinical trial drugs; in process of qualification/validation, application for manufacturing authorisation 2009)

Non-GMP facilities .• Microbial cell culture pilot plant (200 l fermentation)• Animal cell culture pilot plant (70 L bioreactor)• Downstream processing development labs• Analytical labs• Support areas (central autoclaves, cold storage, etc.)


558 m² total area225 m² clean area

Facility GMP II in building Y (1) Basement

BL2 / class C(10 000)

• Microbial Cultivation• Cell Culture• Primary Separation

(centrifugation, homogenisation, micro- and ultrafiltration, depth filtration)

• In-process control lab

BL2 / class D(100 000)


• Media preparation• Cleaning• Purification

(ultrafiltration, chromatography, etc.)

• Cold room processingand storage

376 m² total area342 m² clean area

Facility GMP II in building Y (2) Upper floor

BL2 / EU class C(10 000)

BL2 / EU class D(100 000)


• Multi-use facilities, campaign production

• Easy to clean surfaces (powder-coated steel, stainless steel walls and ceilings, silicone sealed; monolithic pharmaterazzo floor; flush-fitted windows and light units)

• Free space around process equipment affording easy access and cleaning

• Rear-side of cabinets silicone sealed to prevent dust collection

• Highly automated process equipment (low manpower requirements, low human error rate, closed systems)

• Central waste disposal system (automated, coordinated with central CIP system; process streams, floor drains)

Facility design (GMP II):


• 6 air systems:

- Central fresh air supply

- 3 recirculated systems (basement, upper floor class D, upper floor class C)

- Central exhaust air system (filtered for BL2)

- Cold room air recirculation and supply

• 3 level air filtration (F6, F9 and EU13 final HEPA filters)

• Temperature and humidity control

Air handling systems in the GMP II facility:


• Central water supply of deionised water

• Production of purified water by reverse osmosis (approx. 300 kg/h; 3000 l holding vessel; supply ring)

• Production of 1100 kg/h clean steam from deionised water

• Production of WFI from clean steam (approx. 100 kg/h; held at 80°C; 1000 l holding vessel; supply ring)

Water types in the GMP II facility:


• Central Clean-In-Place system (configurable: cleaning agent concentration, temperature, duration, rinse water types [deionised, AP, WFI], optional drying with heated air; ring system supplies all major process equipment)

• GMP grade washer for small articles (stainless steel, glass, plastics; configurable)

• Mobile Clean-In-Place system for other equipment (configurable)

• Central waste disposal system (automated, coordinated with CIP system)

Cleaning process equipment in the GMP II facility:


• Clean room monitoring

- Airborne particulates

- Microbial level (airborne, surfaces)

- Differential pressures

- Temperatures

• Storage

- Refrigerator, freezer temperatures

- Liquid nitrogen tanks (limit alarms)

- Cold room temperatures

• Water quality (cf. European Pharmacopoeia)

- Conductivity

- Microbial contaminants incl. identification of organisms to genus / species level

- Endotoxin

- TOC

- Nitrates

- Heavy metals

Monitoring:


GMP II: Pilot Plant for Production of Biopharmaceuticals for Clinical TrialsBasementBasement: : CultivationCultivation and and primaryprimary separationseparation

65 L Bioreactor, 500 L Bioreactor



Disk Stack Separator



Homogeniser


GMP II: Pilot Plant for Production of Biopharmaceuticals for Clinical TrialsUpper floor: Purification

BioProcess Chromatographysystem


GMP II: Pilot Plant for Production of Biopharmaceuticals for Clinical TrialsUpper floor: Infrastructure

GMP-grade Washer


GMP II: Pilot Plant for Production of Biopharmaceuticals for Clinical TrialsUpper floor: Infrastructure

Chamber Autoclave


GMP II: Pilot Plant for Production of Biopharmaceuticals for Clinical TrialsBasement: Infrastructure

Clean Steam, Purified Water,

Water for Injection


The new aseptic fill & finish unit in Hannover

80 m² clean rooms EU classes D / C / B; class A processing

Approval: Q4/2009 ( scheduled)

Suite 1

Suite 2

D

CB


Quality system requirements

in a development environment


Quality Assurance during Development

Annex 13 of the EU GMP-Guideline states:

• Investigational medicinal products should be produced in accordance with the GMP Guidelines

• Other EU Guidelines should be taken into account where relevant and as appropriate to the stage of development of the product

• The application of GMP is intended to ensure that trial subjects are not placed at risk, and that the results of clinical trials are unaffected by inadequate safety, quality or efficacy arising from unsatisfactory manufacture


Quality Assurance during Development

Annex 13 of the EU GMP-Guideline states:

• Production processes for investigational medicinal products are not expected to be validated to the extent necessary for routine production but

- premises and equipment are expected to be validated

- for sterile products, the validation of sterilising processes should be of the same standard as for products authorised for marketing

- when required, virus inactivation/removal and that of other impurities of biological origin should be demonstrated


EU Regulations

• Directive 2003/94/EG

• EU GMP Guideline (Parts 1 and 2) and Annexes

• EU Points to Consider documents

• European Pharmacopoeia

International

• ICH Guidelines

• CFR / FDA Guidelines

• USP

• PIC/S

German Laws and Regulations

• Drug Act (AMG)

• Drug and API Manufacturing Ordinance (AMWHV)

• German Pharmacopoeia


Inspections

• Local competent authority (in Lower Saxony / Germany this is the StaatlichesGewerbeaufsichtsamt = State Trade and Industrial Inspectorate)

• For certain products (e.g. vaccines, sera, blood products) higher competent authority is the Paul Ehrlich Institute

• For all other products higher competent authority is the Federal Institute for Drugs and Medical Devices

• Inspection by EMEA is required for the central authorisation procedure (to date does not affect us)

Manufacturing authorisation

• Issued by local competent authority

• Novel products will require renewed inspection

• Similar products require notification and documentation

• Original manufacturing license (GMP 1)1999

• Last inspection 2008 with extension of license to GMP 2

• Original manufacturing license (GMP 1)1999

• Last inspection 2008 with extension of license to GMP 2


For products going into clinical trials:

• Investigative Medicinal Product Dossier (IMPD) required (Directive 2001/20/EG).

• GMP requirements apply to all clinical trial phases, but degree and scope of application reflect clinical trial phase

• Examples:

- Cleaning validation not required before Phase III material produced

- Analytical method validation requires

- Validation lots (infamous 3 lots!) required at end of Phase III development

In view of introduction of ICH Q8, Q9, Q10 in EU regulations and the FDA GMPs for the 21st century initiative including the newly updated guideline on validation, this will most probably no longer apply

Division of Pharmaceutical Biotechnology


Head of InstituteProf. Uwe Heinrich

Up-StreamProcessing (microbial)

Dr. Anton Roß

Up-Stream Processing (cell culture)

Dr. S. Duvar / Dr. V. Hecht

Down-Stream ProcessingDr. Jens Paulsen

Quality AssuranceDr. Neophytos Papamichael

Head of DivisionDr. Holger Ziehr

Quality ControlDr. Luma Baydoun

Divisional Organisational Chart / FhG ITEM, Braunschweig Site

Fill & FinishDr. Holger Ziehr


Head of InstituteProf. Uwe Heinrich

Head of ProductionDr. Anton Roß

Head of QCDr. Luma Baydoun

Qualified PersonsDr. Holger Ziehr

Dr. Luma Baydoun

Quality AssuranceDr. Neophytos Papamichael

GMP Organisational Chart / FhG ITEM, Braunschweig Site


Materials Management

• Qualification of vendors

• Testing of materials:

- Raw materials (chemicals, cell substrates [microorganisms / mammalian cells], media; water, steam, gases)

- Process aids with direct / indirect contact with product or critical for correct operation (filters, tubing, chromatography materials)

- Intermediates / product

Released storageRejected storage

Quarantine storage


Quarantine label

Quarantine label

OrderOrder

AnalysisAnalysis

SpecificationSpecification

Supplier

Goods Receipt (QC)Sampling

Goods Receipt (QC)Sampling

Quarantine label

Release label

ReleaseReleaseRejectReject

Materials management

Material transfered to production

area(unopened)

Material used(partially)

Unusedmaterial

destroyed orused in R&D


CAPA SystemCAPA System

Investigation /Final Assessment /

Definition of Actions

CAPA close-out(Documentationand execution

checks)

MaßnahmenauftragMaßnahmenauftragAction disposition

Execution

Deviation Change

Internalinspections

Supplierqualification

OOX

Critical /major

External auditsAction tracking

Qualification /Validation

Production

Other events

Initial Assessment / Definition of Actions

Minor

CAPA: Deviations / Changes / Inspections management

• No Product Quality Review

• Integrated handling


• Qualification and Validation: Major equipment qualification based on risk analysis according to FMEA method (DQ, IQ, OQ, in certain cases PQ). Most important as system components:

- Air handling

- Water and clean steam systems

- Autoclaves

- CIP-System and GMP washer

- T-controlled areas (refrigerators, cold rooms, freezers, liquid nitrogen storage)

• Cleaning verification, validation at Phase III

• Process validation: Validation studies on filtration, virus and endotoxin removal, media fills; reproducibility and consistency; stability


Project development overview and examples


GMP phase of product development is like the tip of an iceberg…

early product development requires more resources in time and manpower!


Costs and time requirements of a “typical” GMP project

Resource requirements

0

5

10

15

20

25

30

35

40

45

Lab scale Scale-up Non-GMP manuf. GMP manuf.

[%]

Project phases

123 6 9 150

Duration (months)

Process development: lab scale

Process development:pilot scale

Non-GMPmanufact.

API-GMPmanufact.

Total approx. 100 – 150 FTE-months per project

Includes analytical development

Transfer and adaptation, e.g. centrifugation steps

Production of reference and non-clinical test substance (PharmTox), product characterisation; initial validation of

analytical methods; initial stability tests

Includes qualification of external laboratories, final validation of analytical

methods

but “typical” is quite variable! Projects can last a few months to

several years

but “typical” is quite variable! Projects can last a few months to

several years

9. Solubilisation

10. Centrifugation

11. Diafiltration

12. Protein chain mix

13. Co-Refolding

14. Depth filtration

15. Concentration/Diafiltration

16. Lactosyl Sepharose

17. Endotoxin reduction

18. Diafiltration

19. Filling

rMLA Inclusion Bodies

9. Solubiisation

10. Centrifugation

11. Diafiltration

rMLB Inclusion Bodies


Project example 1: mistletoe lectin (Aviscumin)

The process at a glance:

• Total of 19 unit operations• Step 1- 8: cultivation, harvesting , cell

disintegration, inclusion body preparation• Step 9-18 purification• E. coli BL 21• 2 x 150 L bioreactor (A-chain, B-chain)• Batch process, IPTG-induction• Batch size: >2 g (API)• Status: API in clinical trials


Project example 2: Erythropoietin (EPO)


• 16 Unit operations• Step 1- 8: cultivation, harvesting,

microfiltration ultrafiltration• CHO• 5 L bioreactor →150 L bioreactor• perfusion process• 9 purification steps• Batch size: > 2g• Status: approved

Depth filtration

Blue Sepharose chromatography

Diafiltration

Q Sepharose chromatography

Filtration

Reversed phase chromatography

MacroPrep HighS chromatography

Ultrafiltration

Superdex chromatography

Filtration

Viral removal (filtration)

Filling

2.5 l perfused concentrate


Project example 3: DNA / plasmids


• 16 Unit operations• Step 1- 4: cultivation, harvesting• E. coli• 150 L bioreactor• batch process• 12 purification steps• Status: clinical trials• Batch size > 1g purified plasmid

Generic API manufacturing process

E. Coli / plasmid cell mass

Resuspension

Alkali lysis

Na acetate Precipitation

Centrifugation

Concentration

CaCl Precipitation

Centrifugation

Diafiltration

Phenyl Sepharose chromatography

Q Sepharose chromatography

UF concentration

Sterile filtration


Project example 4: Bacteriophages MCB Phage stock

Inoculum

Phage lysate

Incubation

Lysis culture

Depth filtration

Sterile filtration

Medium

Sterile filtration• Synthetic Medium: no TSE / BSE risk• Disposables / dedicated use, WAVE®

bioreactor (20l)

• Sterile filtered intermediate / phage lysate

• Reduction of MOI -> increase in phage titer


Project example 4: Bacteriophages Lysate

Ultrafiltration

Bulk - API

Ion exchangechromatography

Size exclusionchromatogaphy

Sterile filtration

Benzonase

330 ± 7,5 nm330 ± 7,5 nm

94 ± 3,0 nm94 ± 3,0 nm (Head)

(Tail)


Clients 1997 - 2009

MediGene AGMunich/Martinsried

Germany

Unilever Biotechnology Application Centre BVBussum

The Netherlands

Dompé Farmaceutici s.p.a.MilanItaly

B.R.A.I.N. GmbHZwingenberg

GermanyMadaus AG

CologneGermany

Fermentas ABVilnius

Lithuania

ZMBHHeidelbergGermany

Mologen AGBerlin

Germany

Viscum AG Zwingenberg

Germany

BiogeneriX AGMannheimGermany

Bioceuticals Arzneimittel AGBad VilbelGermany

Hexal Biotech ForschungsGmbHHolzkirchen

Germany

F. Hoffmann La-Roche Ltd.Basel

Switzerland

PolyPhag

PolyPhag GmbHViersen

Germany


Outlook


Fraunhofer ITEM objectives:

• Single-stop partner for development, production, pre-clinical and clinical testing of (respiratory) medical products

• Optimisation of microbial fermentation and mammalian cell culture as primary manufacturing routes, efficient downstream processing

• Production of APIs and final dosage forms for clinical trials tointernationally recognised GMP quality levels

• Standardisation of production vehicles – antibody/antibody fragments and DNA/plasmid production platforms for speedy first in human trials

• Consultancy services in GXP


Thank you

for your attention

Documents

Operation of a pilot plant for clinical lots of ...Operation of a pilot plant for clinical lots of biopharmaceuticals Client types: • Academic institutions (1990’s) – starting