Manufacture of Somatic Cell Therapy- and Tissue-Engineered

Section 6/3 - R. Sanzenbacher

Manufacture of Somatic Cell Therapy- and Tissue-Engineered Products: Considerations on Quality Aspects

Disclosure: I attend this conference as an individual expert. Some views expressed here are my personal views, and may not be understood or quoted as being made on behalf of the PEI or EMEA or reflecting the official position of those.

ISCT Rotterdam, 21. May 2011

Cell-based products: Manufacturing and Control The key material: Cells as starting material and active substance EU Guidance: Overview

http://www.celltherapysociety.org/Default.aspx


Procedures for Tissue/Cell-Products at PEI Div 6 (2004- 2Q/2010)

(without Therapeutic Tumor Vaccines) (without non-manipulated HSC for heterologous use)

61 26 42 44

2010 (2Q) 2009

2004 2005 2006 2007 2008

centralised national

0

5

10

15

20

25

5

18

4 2

1

3

7

1

10


Quality Aspects of Cell-based Medicinal Products (CBMPs)

Transport

Manufacturing

Formulation for Administration

Transport Testing

Release Storage

Procurement

Active Substance

Drug Product



Manufacturing of Cell-based Products: Qualification / Validation

Transport

Manufacturing

Formulation

Transport Testing

Release Storage

Procurement

Active Substance

Drug Product


Starting materials Donor Qualification

Staff Training Transport

Materials, Reagents, Exipients

Process Product Characterisation

In Process controls

Testing

Release Specifications

Transport Stability

Packaging materials

Labelling

Traceability

Administration

Process Variations

Handling at clinical site: Staff Training

Procurement Process

Standardisation

All steps have potential impact on the quality/safety of the product !


human tissues/cells for human application

incl. SCs excl. blood blood components

National Accreditation System

Dir 2004/23/EC Processing, Preservation, Storage, Distribution,…

Tissues, Cells

autologous grafts used within same surgical procedure

(parts of) organs for homologeous use

Dir 2004/23/EC Donation

Procurement Testing

Reproductive cells

Dir 2006/17/EC

Dir 2006/86/EC

Regulatory Environment in the EU

not “prepared“

Tissue Preparations



human tissues/cells for human application

incl. SCs excl. blood blood components

National Accreditation System

Dir 2004/23/EC Processing, Preservation, Storage, Distribution,…

Tissues, Cells

autologous grafts used within same surgical procedure

ATMPs Centralised MA Medicinal Products

Organs/ parts of organs for homologeous use

Dir 2004/23/EC Donation

Procurement Testing

Reproductive cells

Dir 2006/17/EC

Dir 2006/86/EC

CBMPs: Regulatory Environment in the EU

Reg (EC)1394/2007 on ATMPs

Dir 2001/83/EC as amended

not “prepared“

National Accreditation (Hospital Exemption)

Tissue Preparations



ATMPs: Current EU Guidance

✘ 2008: Potency testing of cell-based immuno- therapy MPs for treatment of cancer Doc Ref: CHMP/BWP/271475/06 ✘ 2008: Guideline on Human cell-based MPs Doc Ref: CHMP/410869/06 ✘ 2010: Reflection paper on Chondrocyte containing MPs for cartilage repair Doc Ref: CAT/CPWP/568181/2009 ✘ 2010: Guideline on Xenogeneic CBMPs Doc Ref: CHMP/CPWP/83508/09 ✘ 2011: Reflection paper on stem-cell based MPs Doc Ref: CAT/571134/09

http://www.ema.europa.eu/ema


✘ PART IV: Advanced Therapy Medicinal Products

A risk-based approach may be applied to determine the extent of quality, non-clinical and clinical data to be included in the MAA

Special provisions applying for ATMPs ✘ PART I: (Section 3.2)

The Pharm. Eur. monographs shall be applicable to all substances, preparations and pharmaceutical forms appearing in it. Reference shall be given.

In the case of analytical procedure(s) included in the Pharm. Eur., the description shall be replaced by the appropriate reference to the monograph(s) and general chapter(s).

Where starting and raw materials, active substance(s) or exipient(s) are not desribed in a Pharm. Eur. or a EU MS monograph, compliance with a monograph of a third country pharmacopoeia can be accepted.

In case where a specicification in a Pharm. Eur. or in a EU MS monograph might be insufficient to ensure the quality of the substance, the CA may request more appropriate specifications. Where applicable and if needed, a certificate of conformity (CE marking) on medical devices shall be provided.

ATMPs: Dir 2001/83/EC, Annex 1


✘ 2.6.1. Sterility (Guideline for using the test in in 5.1.9)

✘ 2.6.7. Mycoplasmas

✘ 2.6.14. Bacterial Endotoxins (GL for using the test in 5.1.10)

✘ 2.6.30. Monocyte Activation Test (alternative to rabbit pyrogene test)

✘ 2.7.24. Flow Cytometry

✘ 5.1.6. Alternative Methods for Control of Microbiological Quality

✘ 5.1.7. Viral Safety

(✘ 5.2.3. Cell Substrate for Production of Vaccines)

✘ 5.2.8. TSE

✘ 2.6.27. Microbiological Control of Cellular Products

✘ 2.7.24. Flow Cytometry

✘ 2.7.28. Colony-forming Cell (CFC) Assay

✘ 2.7.29. Nucleated Cell Count and Viability

✘ 01/2008:2262 Bovine Serum

✘ 01/2008:2323 Human Haematopoietic Stem Cells

CBMPs: Relevant Ph.Eur. Chapters and Monographs




CBMPs: Quality Aspects THE MAJOR VARIABLE: Tissues/Cells as starting material

Transport


Transport Testing

Release Storage

IPCs Variations

Procurement Starting Material

Manufacturing


✘ Origin and banking history of cell lines (History Files, )

✘ Definition of (primary) master/working bank

✘ Performance

✘ Comparability of performance over time (passage numbers, storage)

✘ Phenotype, identity, purity

✘ Genetic stability, karyotype

✘ Virus screening (serology, PCR, indicator cell lines): HIV-1/2, HBV, HCV, HTLV I/II, EBV, CMV, Parvovirus B-19, ..)

✘ Adventitious agents screening (Bovine, mouse, porcine, rabbit; ampho-/xeno-/ecotropic retroviruses,…)

✘ Microbiological screening

Starting Material – Qualification of Cell Banking System CBMPs Quality Aspects

References: EP 5.2.3. Cell substrate for Production of Vaccines ICH Q5a (=CPMP/ICH/295/95/ICHtopicQ5a) ICH Q5d EudraLex Vol 4 (GMP), Annex II



CBMPs Quality Aspects Starting Material – Control of a Primary Cell Source

✘ Qualification of Donor - Intensive donor screening must substitute for virus inactivation ✘ Control of Procurement - Standardized procurement of cells/tissues - Training of procurement team - Prevention/Reduction of microbial contamination risk at procurement site ✘ Control on Biopsy Transport and Receipt - Stability considerations (time, temperature, stress conditions,. to preserve functional integrity) - Risk of microbial contamination - X-linking of different traceability systems (30y!!!)



Starting Material from Human Source: Viral Safety Principles

Donor Selection Donor Testing Inactivation/

Removal Vigilance / Traceability

Donor compliance Unknown risks Limited tissue resources

Lim

itatio

ns

Unknown viruses Sensitivity Diagnostic window Test interference post-mortem donor: Limited Shelf life for further preparation

Product integrity Virus stability (Residual contam.)

Compliance Detection

Definition of exclusion criteria Donor evaluation: Medical history Behavioural History Physical examination

Procurement report

ELISA PCR

Filtration Thermodesinfection Pasteurisation Chemical (PES,..) Gamma-irradiation

QM-Systems RMP

Met

hods



Human Donor Testing

Dir 2006/17/EG Annex 2 Other Guidance, eg. for blood, blood components , blood-rich tissues NAT might be applicable Additional product-specific testing might be requested e.g. ParvoB19, WNV, CMV …



Transport


Transport Testing

Release Storage

IPCs Variations

Procurement

Manufacturing

CBMPs Quality Aspects


Sterility of starting material can not be guaranteed Compare to transfusion medicine where 0.2 to 0.5 %

of cellular blood components (Thrombocytes) are bacterially contaminated,

e.g. Staph. epidermidis (Skin as tissue source)

Increasing incidence of multi-resistent hospital strains

(MRSA, Acenetobacter, vancomycin-resistent Enterococci)

Non-sporocidal disinfection procedures

Starting material cannot be sterilised Final product cannot be sterilised e.g. heat, gas, γ-rays, filtration, 0.2 µm filters

Use of antibiotics mask bacterial contamination, concentration might be lower in patient

CBMPs Quality Aspects Microbiological Safety


Sterility testing underlay “sampling error” ✘ The potential result “The test sample is sterile” gives no significant information on the whole volume of product/intermediate ✘ Antibiotics in culture medium disguise contamination

Short shelf life of final product ✘ Testing results not available at release / application to patient


Sterility (EP 2.6.1) Method: Membrane filtration or direct inoculation Readout: Turbidity Incubation time: at least 14 days at 30-35°C

Microbiological Control of Cellular Products (EP 2.6.27) Readout: CO2 Production Incubation time: at least 7 days at 35 - 37°C

BacT/Alert, BioMerieux

BACTEC, BD Diagnostics

Established Methods for Sterility Testing might not be appropriate ✘ Media, Temperature, Interference


Consequences

✘ Implementing of a complex overall microbial safety strategy for cell products

✘ Development of rapid test systems or combination of systems

(PCR, FACS, Substrate, Bioluminescence,…)

✘ If justified by product and indication, CA might accept test limitations

✘ Need for Revision of Guidelines and Monographs

References: EP 2.6.30 Monocyte Activation Test (Pyrogen Testing) FDA 2008 Guidance for Industry: Validation of Growth-Based Rapid Microbiological Methods…



✘ limited amount as starting and test material

✘ active substance often “cell mixtures” where therapeutic effect cannot be related to single cell type or defined combination of cell types

✘ inherent variability within one cell type (donor age, gender, ethnicity, disease state, comorbidities,…….)

✘ heterogeneity of different cell types upon culture, eg dedifferentiation, genetic instability, malignant transformation,..

✘ fragility ✘ undefined mechanism of action(s)- Controls?

✘ cells interact with/depend on (micro)environment

✘ cells “biodistribute” actively

✘ …..

© Steve Sack -The Minneapolis Star Tribune

CBMPs Quality Aspects Cells as Complex Active Substance


Keep in mind: Potential Safety Issues associated with CBMPs

Infections / Microbial Contamination Inflammation Immunogenicity, Rejection Immunosuppressive Action Inappropriate differentiation loss of function Malignant transformation, Tumourgenicity Ectopic engraftment to non-target tissues

Cancer Res 2005

Jan 2008


CBMPs Quality Aspects Cells as Starting Material and Complex Active Substance

Consequences for process development

✘ Due to inherent variability of (primary) cells, variability in some process steps and respective specifications ranges should be included ✘ High impact of process changes /up-out-scaling/ other deviations on products should be considered already in early product development ✘ Biological analytical techniques show a greater variability than physico-chemical tests

✘ For reproducibility other variables must be reduced and/or controlled by QC measures as best possible -staff training -adequate tools to demonstrate lot-to-lot consistency and comparability (relevant IPCs and validated robust accurate tests) -defined quality of reagents and materials -quality risk management system, risk profiling strategy


CBMPs: Influence of Ancillary Materials

Morphology Metabolic activity

Gene Expression Profile

Proliferation

Intracellular Signalling

Differentiation

Extracellular Signalling

Protein Distribution Profile

Secretory Activity Vitality

Migration, Adherence

Processing Steps

Cell functionalities (mode of action + others), stability, safety

Starting Material

Reagents & Materials

??? ???

Culture Conditions


CBMPs Quality Aspects Product Characterisation / Release Specifications

✘ ✘ Total Cell Number ✘ ✘ Cell Viability ✘ ✘ Identity (phenotypic and/or genotypic) ✘ ✘ Purity ✘ ✘ Impurities (product-/process-related, degradation products, adventitious agents) ✘ ✘ Potency ✘ ✘ Sterility ✘ Structural Characteristics (3-dimensional shape) ✘ Senescence


Definition of combined ATMP

• ‘Combined advanced therapy medicinal product’ means an advanced therapy medicinal product that fulfils the following conditions: – it must incorporate, as an integral part of the product, one or

more medical devices within the meaning of Article 1(2)(a) of Directive 93/42/EEC or one or more active implantable medical devices within the meaning of Article 1(2)(c) of Directive 90/385/EEC, and

– its cellular or tissue part must contain viable cells or tissues, or

– its cellular or tissue part containing non-viable cells or tissues must be liable to act upon the human body with action that can be considered as primary to that of the devices referred to.

Art 2(d) of Reg. 1394/2007


Combined ATMPs are evaluated by EMA as a whole, since the agency is responsible for the final decision and control on the medicinal product

MAA should include results of the NB’s report on the device part in section 3.2.R of CTD; the agency shall recognize this results in its evaluation CAT may demand transmission of evaluation results of the medical device part by a NB If no results are available at time of MAA, CAT shall seek an opinion of a NB which is to be determined together with the applicant However, involvement of a NB is not necessary if the CAT believes to have sufficient expertise

Combined ATMPs: Consultation of Notified Bodies (NB)


Ralf Sanzenbacher Paul-Ehrlich-Institut Federal Insitute for Sera and Biomedicines Division of Medical Biotechnology Section Tissue-Engineering & Somatic Cell Therapeutics D-63225 Langen/Germany E-mail: [email protected]

Thank you! MAA

accepted

Manufacture of Somatic Cell Therapy- and Tissue-Engineered Products: Considerations on Quality Aspects

mailto:[email protected]

Documents

Manufacture of Somatic Cell Therapy- and Tissue-Engineered