cGMP Production on RFH Campus

Dr Mark W LowdellDirector of Cellular Therapy

Royal Free HospitalUCL Medical School

London, UK

Paul O’GormanLaboratory of Cellular Therapeutics

Royal Free Hospital

How are cell therapy trials regulated?

• Pre 2004– 1995 Council of Europe begins proposals for regulation of clinical cell &

tissue therapies– 1997 UK DoH publishes CoP and voluntary accreditation scheme via MCA

(MHRA)– 1999 POG LCT accredits to voluntary scheme– cGMP not required; proposal for EU GTP standards akin to FDA

• 2004 Clinical trials directives enacted – include “substantially modifiedsomatic cells” as IMP for the first time. cGMP manufacture requiredplus MA (IMP) and Qualified Person.

• 2007 First LCT product to be deemed an IMP by MHRA

• 2009 ATMP Regulations published– Procurement of starting material regulated by HTA and requiring licence– “nonsubstantial” defined– Inclusion of HEC for one-off, non-trial products

How are cell therapy medicinalproducts regulated?

• Investigational: MA(IMP) and QP required– Clinical trials directives for conduct (GCP)

– Annex 1-13 of medicinal products directives

– ATMP regulations 2009 for manufacture (cGMP)

– HTA Quality & Safety Regulations

• Non-investigational: MA(Specials) (or HES) and QC required– Annex 1-13 of medicinal products directives

– ATMP regulations 2009 for manufacture (cGMP)

– HTA Quality & Safety Regulations

Both require full cGMP production

cGMP requirements

• cGMP production– Document control

– Staff training

– Materials qualification, control & tracking

– Equipment qualification and validation

– Environmental control & monitoring• Air borne particles (in process and at rest)

• Viable particles (settle plates, finger dabs and product testing)

• “Open” –

– A in a B background (A in C in US)

– In isolators in D background

• “Closed” – D environment

– Released products control & tracking• AR/SAR reporting

Paperwork more important than facility

How did we meet cGMP at RFH?

• New labs designed in 1999 to GTP standards in US– Grade C background– No Grade B gowning required so single stage change

• Opened in 2000• 2004 apparent that cGMP standards will be applied in UK

– Grade B background needed• Increased air volume exchange – no recirc possible• Increased differential pressures – increased fan speeds

– Grade B gowning• Stage 1 change area needed

– BUT:• cGMP gowning not compatible with human tissues (HOWIE rules)• Wash hand basin required in de-gowning area

– Facility design imperfect for minimisation of X-contamination• Process design to overcome limitations of facility design

EU vs FDA for “open” processes

EU cGMP requires ISO5 in ISO5 background (at rest) but ISO7 background in operationFDA cGMP requires ISO5 in ISO7 background at rest and in operation (effectively a “C”)

3,500,000

3,500,0002,000350,000

2,000350,00003,500

03,50003,500

5m0.5µm5m0.5µm

GradeMaximum permitted number of particles / m2 at or above size specified

A ISO5

B ISO5

C ISO7

D ISO8

At Rest In Operation

20,000

20,000

Not Defined Not Defined

How did we meet cGMP at RFH?

Stage 2Change

Class C

Stem cell and pre-IMP processing LaboratoryLab 1 - Class B

La

b2

-C

lass

B

Lab 4 - Class B

CO2 incubator

Fridge / freezer

Transfer hatch

Bench

Critical work area

(under bench)

Class II microbiological cabinetClass A

Wash hand basin

28.26m3

40.8m3

18.0m3

15.9m3

8..8m3

Lab 3 - Class B

StoreRoom

Quarantine

LockerRoom

&Stage 1Change

Class C lab

The RFH cGMP facility

Lab 2

Lab 3

Lab 4

How did we meet cGMP at RFH?

• Dual standard operation– Non-cGMP production

• Labs operated to Class B air standard BUT …• Gowning to Class C standard

– cGMP production• All labs operate to cGMP with full class B gowning

• Campaign use of Labs– Lab 1 is preparation lab for starting material

(the only lab to be used for non-cGMP work)– Lab 2 is preferred lab for short process cGMP manufacture– Lab 3 is preferred lab for mid length cGMP manufacture (2 days)– Lab 4 is the only lab for long-term cGMP manufacture (lines) and

for islet cell production (most “open” procedure currently)– No access to Lab 4 when Lab 3 is in operation and vice versa

Gowning and access

• Non-cGMP production – “A” in a “D” background

Sterile docker used to makemost processed “closed”Although some are “open”

Gowning and access

• cGMP production - “A” in a “B” background

Our first cell therapy IMP - TaNK

• Pre-clinical testing

– Regular research data repeated to GLP standards

2.01.3520.0010

0.40.286.009

2.21.4710.108

2.71.8416.707

4.42.9923.006

3.02.0224.905

2.01.3614.504

2.11.4128.903

4.02.7337.902

3.62.4525.901

x10^7/kgx10^9x10^9Donor

CD56CD56MNC STABILITY OF CTV-1 PHENOTYPETHROUGH 28 MEDIA CHANGES

0 2 4 6 8 10 120

5

10

15

20

CD2

CD3

cCD3

CD4

CD5

CD7

CD8

CD15

CD19

CD33

CD34

CD56

HLA-DR

70

80

90

100

Week

%P

OS

ITIV

E

RPMI +10%fcs XVIVO-100.0

2.5

5.0P1

P2

P3

P4

P5

P6

Comparison of CTV-1 proliferation in TC flasks (RPMI+10%FCS )with cells cultured in T-cell expansion bag in X-VIVO 10

Frozen vials of CTV-1 from same previous passage were thawed and cultured in either RPMI+10%FCS

or X-VIVO 10. Initial cell conc. for both 0.5x106/ml. Cells were split 50:50 every 48hours (except P5 were cellsfed after 72hrs)with fresh medium and cell conc. checked.

Ce

llc

ou

nt/

ml

Our first cell therapy IMP - TaNK

• Pre-clinical testing

– Regular in vitro research data repeated to GLPstandards

– No animal model (xenogeneic system inappropriate forsafety or efficacy – e.g Tegenero)

Our first cell therapy IMP - TaNK

• Pre-clinical scale-up

– Anti-CD56 selection reagent

– Immunomagnetic sorting system

Our first cell therapy IMP - TaNK

• Pre-clinical scale-up problems– Validation

• Product definition? (e.g. MSC)

• Gold Standard reference analyte?

• Validated GLP assays?

• GLP labs?

– PSF design• Starting materials are human cells and thus highly variable

• Functional assays impossible to validate to usual pharmaceuticalstandards

Our first cell therapy IMP - TaNK

• Manufacturing for phase I/II trial

– Release criteria• Lack of reference analyte

• Highly variable functional assays

• In vitro potency assays not available on thawed product

• Viability not available on thawed product

• Sterility not available on fresh product

Our first cell therapy IMP - TaNK

• Current status

– Trial commenced 10 months late

– 14 patients enrolled and 8 treated in 18 months (1 offtrial)

– No deaths in treated group

– 7/8 patients responded to treatment

– Trial closed

Grade BLab 1

Grade B corridor

Grade BLab 2

Grade BLab 3

Grade BLab 4

Male

sta

ge

1

Fem

ale

sta

ge

1

Grade DT&CD processing

&QC

CL IICL II

CL

II

CL II

CL IICL II

Gene Rx culture lab

Gene Rx preparation lab

CO

2

CO2 CO2 CO2 CO2

CO2 CO2 CO2 CO2

CO

2+

4C

+4C

CLII

CO2 CO2 CO2 CO2

+4C

CL

IIC

LII

Stage 2 change

CL

IIC

LII

Gra

de

CT

&C

Dp

rocessin

g

+4C

+4C +4C +4C

+4C

+4C

Grade C Lab

Qu

ara

nti

ne

Go

od

sS

tore

LN2CO2 CO2 CO2 CO2

Grade B -ve

Grade B -ve

AirLock

Planned cGMP facility for RFH

Conclusions

• Increasingly we will have to move to full cGMP compliance

• This is less about facilities than systems

• Process can be used to overcome inadequacies of design

• Pragmatism is the order of the day – don’t “gold plate” theregulations

• BUT ………

Sometime even the best process can’tovercome the design!!!