14 October, 2010 Regulatory Challenges for Human … challenges... · • Cell manufacturing requires special training, facilities, and ... XF vs XF+LM. 10 ... • 510(k) cleared

Regulatory Challenges for Human Cell and Tissue-Based Products:

Considerations from the Tool Provider’s Perspective

Mark Bonyhadi, PhDDirector, Clinical Business DevelopmentCell Therapy

2010 Taipei International Symposium on Human Cell and Tissue-based

Products and Tissue Banks14 October, 2010

2

Disclaimer

• This presentation is for your general knowledge and background only. The presentation includes information from various sources considered to be dependable. However, we make no representations, warranties or other expressed or implied warranties or guarantees regarding the accuracy, reliability or completeness of the information. Proper attribution should be provided for any use of non-public information contained in this presentation. The information provided does not constitute legal advice. You must consult with competent legal counsel for legal matters. You are fully responsible for complying with all applicable regulatory and safety requirements associated with the products discussed in this presentation. Under no circumstances shall Life Technologies be liable for damages or losses or any kind that may arise from your use of the information in this presentation.

3

Expand Qualify StoreIsolate

Growth factors& substrates

ELISA

Cell selectionbeads

Cell imaging

Gene expression

Media, supplements& single use

processing systems

Cryopreservationmedia

Flow cytometry

Monitor

Dissociation enzymes

Processingdevices

Cell Therapy Workflow Solutions

HLA typing

Immuneresponse

monitoring

4

Cell Therapy’s Challenges in an Evolving Space: Why is cell therapy so unique?

• Cells are unlike conventional drugs− Complex - Persist in vivo− Change over time - Multiple modes of action− Source varies (donor, tissue, age)

• Traditional models for testing safety/toxicity may not apply

• They and/or the process for their manufacture are targets for infection and/or contamination− They cannot be terminally sterilized

• The tools and methods for characterizing, selecting, processing,growing (stem) cells are still evolving

• Cell manufacturing requires special training, facilities, and compliance

5

Safety Issues: Learning Things the Hard Way

October 20, 2003

Gene therapy 'caused T-cell leukemia'Insertional mutagenesis pinpointed as cause of T-cell leukemia in X-SCID gene therapy trial | By Jo LyfordAn unanticipated complication of gene therapy has been confirmed as the cause of T-cell leukemia in two boys receiving the pioneering treatment for X-linked severe combined immunodeficiency (X-SCID). In the October 17 Science, an international research team confirms that inappropriate insertion of the retroviral vector near the proto-oncogene LMO2 promoter led to uncontrolled clonal proliferation of mature T cells (Science, 302:415-419, October 17, 2003).“This is the nightmare scenario,” said Terry Rabbitts of the Medical Research Council's Laboratory of Molecular Biology in Cambridge, whose previous work on LMO2 played a key role in the investigation. “It's time to step back and give ourselves some breathing space.”

6

Safety Issues: ESC/iPSC - TeratomasThe inherent multipotential nature of ESC/iPSC carries risk of causing teratomas and/or innapropriate differentiation

pp. 743-745

7

Safety Issues: ContaminationThe culture process for generating cells, as well as the quality of the (raw) materials used to isolate, select, culture and expand (stem) cells for therapy is a potential source of contamination

8

Safety Issues: Mitigating risk through new technology

• Evolution of new reagents, tools, and instrumentation appropriate for cell-based therapies

1. Reduce risk of contaminationRapid contamination evaluationTransition to “safer” reagentsRemoving undifferentiated cellsImproving “rare event” detection

2. Expanding cell characterization capabilitiesGetting real-time data from precious samplesMaximizing data capture: amount and kinds of data

3. GMP cell manufacturingAutomationClosed systemsMonitoring/sampling

9

Transition to safer reagentsAnimal-Origin-Free

Xeno-FreeSerum-Free

Serum-containing

FBS, MSC-Qualified

StemPro®MSC SFM

StemPro®MSC SFM XenoFree

Next Generation

Von Kossa (Calcium Deposit)

Alkaline Phosphatase

Alcian Blue (Proteoglycan)

Oil Red O (Lipid Vessible)

Osteogenesis

Osteogenesis

Chondrogenesis

Adipogenesis

Day 12 Expanded MSCs

MSC Isolation and Expansion

Data generated by Christian Prante, Data generated by Christian Prante, Wolfgang Wolfgang ProhaskaProhaska and Knut and Knut KleesiekKleesiek --

RuhrRuhr--UniversitUniversitäätt BochumBochum

Passage 9Passage 9

• Recombinant AOF Enzymes• Xeno-Free Substrates• Mf in GMP Facilities

Population doublings for ADSC: Classic media + FBS, XF vs XF+LM

10

Rapid contamination evaluationMycoplasma Detection Assay: MicroSEQ® Assay• An integrated sample preparation

and real-time, quantitative PCR assay for the detection of Mycoplasma in cell culture samples

• Rapid sample preparation and same-day results allow for in-process testing

10 copies

1 copy

Improving “rare event” detectionAttune™ - Acoustic Focusing Cytometer• Faster sample acquisition times (10x)

• Can use dilute samples/no-wash techniques

• Absolute cell counts with external counting reference (e.g. beads)

• Allows for rapid “rare-event”detection/enumeration

11

Removing undifferentiated cellsCASE IN POINT:

Cellular therapies derived from hESC:Considerations for Pre-Clinical Safety testing and Patient MonitoringCellular, Tissue and Gene Therapies Advisory Committee, CBER, FDA, April 10 2008“Cell therapy products: Adopt manufacturing practices that minimize the number of undifferentiated hESC present in the final formulated preparation”

Neural stem cells derived from hESCcontain undifferentiated hESCBead Based Cell

Separation Systems

Dynabeads®

SSEA

-4 A

PC

0.2 %14 %SS

EA-4

APC

Pre-SSEA4 Bead

Depletion

Post-SSEA4 Bead

Depletion

12

Expanding Cell Characterization Capabilities

Massively parallel next-generation sequencing platform that supports a wide range of applications, with multiplexing capability allow multiple experiments in a single run.

• Safety is a major challenge for cell-based therapies• Cells go through multiple passages and are in culture for long periods of time

Concerns:Genomic stabilityEpigenetic integrityGene-Rx insertional effects

GenomeDe Novo SequencingTargeted ResequencingWhole Genome ResequencingEpigenomeChromatin Immunoprecipitation SequencingMethylation AnalysisTranscriptomeGene Expression ProfilingSmall RNA AnalysisWhole Transcriptome Analysis

SOLiD™ System with Barcoding technology

13

Methylation-specific castPCR• Detection of Rare Undifferentiated hESCs• Detection of Specific Lineage Methylation Signatures

Product purity/differentiation potential

14

Development & Regulatory Challenges facing Cell Therapy Developers

• Pre-clinical data generation

• IND filing

• Clinical manufacturing/trials

• BLA

All require demonstrated phase appropriate risk control for reagents selected as part of the Chemistry, Manufacturing and Control

(CMC) process

15

Reagents for the Cell Therapy Work Flow

Tissue dissociation enzymes

Culture medium

Cryopreservation solutions

Serum

Growth factors

Antibiotics

Monoclonal antibodies

• You must list in your IND any reagents used in manufacturing the “product”• Reagents are materials that are used for:

• Cellular growth

• Differentiation

• Selection/depletion

• Purification

• Other critical mfg steps, but are not intended to be part of the final product

• All can affect the safety, potency, and purity of the final product (e.g. introduction of adventitious agent)

16

Challenges include:

• Variability and complexity inherent in the components used to generate the final product

• Source of cells

• Potential for adventitious agent contamination

• Aseptic processing

• Inability to “sterilize” the final product

17

Control of Raw Material Quality

• Manufacture of a cellular product of defined quality relies on thorough description, characterization, and testing beginning with:

Source MaterialsReagents Ingredients Components used throughout the manufacturing process.

• Contingent upon developing a qualification program implemented during product development: applies to raw materials used to manufacture product.

18

“Tabulation of Reagents Used in Manufacture”• Recommended to provide the following information on

“ALL” reagents used during product manufacturingConcentration of reagent at each manufacturing stepVendor/supplierSource:

Human: licensed product, clinical or research gradeCOA or info regarding testing of donor and/or reagent

Porcine:COA or documentation that product is free of porcine parvovirus

Bovine:ID Bovine material and sourceLocation and history of herd

19

Reagent Quality

• Recommended to use FDA-approved or cleared, GMP manufactured, or clinical grade reagents whenever they are available.

• COA or cross-reference letters

• USP grade

• Even FDA-approved materials may need additional testing

20

Reagent Quality

• If a reagent is not FDA-approved or cleared (e.g. research grade):

− Identify source of material− Additional testing may be needed− Implement a qualification program to provide info in IND

Safety testingMycoplasmaAdventitious agentsPurity testing

Sterility EndotoxinPerformanceAssays (e.g. residuals)

• Recommended to use FDA-approved or cleared, GMP manufactured, or clinical grade reagents whenever they are available.

• COA or cross-reference letters

• USP grade

• Even FDA-approved materials may need additional testing

21

More things to think about…

• Pay attention to reagent labeling

− Materials designated RUO or other− Animal Origin Free − Xeno-Free− IVD− FFM− For manufacturing of cell-based products

• Pay attention to quality systems under which reagents are manufactured− Produced under GMP vs. ISO

22

• 510(k) cleared products

• Drug Master File

• Raw Materials Specifications

• Raw Material Suppliers Qualifications

• Regulatory Affairs Support

• Compliance with 21 CFR Part 820

• ISO 13485:2003 Certification

Reagent selection criteria to help reduce regulatory burden for Cell Therapy Developers:

23

• 510(k) cleared products

• Drug Master File

• Raw Materials Specifications

• Raw Material Suppliers Qualifications

• Regulatory Affairs Support

• Compliance with 21 CFR Part 820

• ISO 13485:2003 Certification

Reagent selection criteria to help reduce regulatory burden for Cell Therapy Developers:

24

Drug Master File

• DMF ContentsProcessing and Production MethodsRaw Material SpecificationsProduct FormulationsIncoming Quality Control and Finished Product Release TestingQuality Assurance Oversight and Batch Record Review and Release

• Information contained in the DMF may be used to supportInvestigational New Drug Application (IND)New Drug Application (NDA)Abbreviated New Drug Application (ANDA)Amendments and supplements to any of these.

25

Raw Material Specifications

• USP/EP requirements where possible− If a USP/EP grade material is not

available then an alternate source such as: America Chemical Society (ACS), Food Chemical Codex (FCC), or internal specifications can be developed to ensure raw materials meet the required purity appropriate for that material.

• Certificate of Analysis

• Certificate of Origin

26

Raw Material Supplier Qualifications

• Audits to both ISO 13485 standard and 21 CFR Part 820

• Agreements to ensure:− Continuity of supply− Notification of changes

• Manufacturers must work with suppliers to identify − primary/secondary/tertiary components used in

manufacturing− identify animal components or materials that may not

be conducive to cell therapy applications

Need controls to enable confidence in supply of quality goods

27

Animal-Derived Materials

• Eliminate if possible. Explore recombinant or non-animal-derived alternatives. − Bovine serum, porcine trypsin, other enzymes, collagen…

• Establish freedom from species-specific viruses. Other precautions -closed herds, specific countries of origin− Requirements for ingredients of animal origin used for production of

biologics (9 CFR 113.53)

• Bovine-derived materials - potential risk of transmissible spongiform encephalopathy (TSE)− Proposed Rule: Use Of Materials Derived From Cattle In Medical Products

Intended For Use In Humans And Drugs Intended For Use In Ruminants, January 2007

• Investigate the raw material of the raw material. − Bovine material used upstream in raw material manufacturing?

28

Qualification Testing

• Performed to assure quality of raw material received from manufacturer− In addition to testing reported on raw material COA

• Risk-based approach to qualification testing− Intrinsic risks of material− Results of supplier qualification - balance supplier’s limitations− More extensive testing needed for higher risk materials, or to compensate

for supplier limitations

• Testing program - in-house, or (qualified) contract testing lab

29

Materials Qualification - a Risk-Based Approach(USP Chapter 1043)

• Tier 1: low risk, highly qualified− HSA, insulin− Certificate of Analysis, assess removal from final product

• Tier 2: low risk, well-characterized, GMP-manufactured, used as ancillary material, not animal origin− Growth factors, density gradient medium− Add supplier qualification

• Tier 3: moderate risk; not intended for use as an ancillary material, diagnostic or research grade (growth factors, culture medium)− Additional testing needed for qualification

• Tier 4: high risk, potentially toxic, animal-derived (feeder cells, FBS)− Source animal, documentation of country of origin

30

Supplier Regulatory Affairs Support• Manufacturers should have expertise with FDA

and foreign regulators.

• RA professionals should have experience in the following areas:− 510(k) submissions− PMA submissions− EU CE Mark Technical Files− Submissions in LATAM and ASIA/PAC − cGMP Compliance − Knowledge of products and how they may be

used in both clinical and research applications

31

Summary• There are many new “innovations” that can be used to

enhance to safety of cell- and tissue-based products

• There are many tools that can used to help reduce the regulatory burden to cell therapy manufactures− 510(k) Cleared Products

− Drug Master File

− Raw Materials Specifications

− Raw Material Suppliers Qualifications

− Regulatory Affairs Support

− Compliance with 21 CFR Part 820

− ISO 13485:2003 Certification