The Development and GMP manufacture of adoptive T cell therapies

Ryan Guest University of Manchester – Cellular Therapeutics Unit

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Overview

• T cell immunotherapy overview

• Origin of the University of Manchester CTU

• ATMP products & current clinical trials and treatments

• Associated toxicity & Challenges for the industry

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T-cell receptor or CD3 complex

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Vα

Cα

Vβ

Cβ

CD3ζ homodimer

α - chain & β - chain heterodimer

CD3-γ,-ε heterodimer

CD3 -ε,-δ heterodimer

Cytoplasmic signalling domain

T cell membrane

Variable binding domain for MHC-antigen recognition

Processed Tumour peptide in MHC

Antigen presenting/Tumour cell membrane

T-cell receptor & MHC complex

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Vα

Cα

Vβ

Cβ

Induces T cell stimulation

T cell membrane

Tumour cell membrane

Collect T-cells from Patient

Expansion of tumour specific T cells

T-cells returned to Patient

Adoptive T cell Therapy for Cancer

(1) Natural anti-tumour

T cells

(2) Engineered anti-tumour

T cells

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1 - Natural anti-tumour T cells for Therapy

Excised tumour

Rapid expansion

Re-infusion to conditioned

patient

Proliferation of lymphocytes infiltrating tumour driven

by cytokines

Tumour Infiltrating Lymphocytes

Cells frozen after culture (< 30 days)

Cells defrosted

TIL - 1010-1011 cells

TIL - 107-108 cells

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2 - Gene engineered - T cells for Therapy

T-cell receptor gene therapy

Recombinant TCR α & β

Antibody targeted T-cells

CAR

Chimeric Antigen Receptor

scFV (single chain variable

fragment)

CD3 Signalling domain

MHC + epitope

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± co-stimulatory

domains

Collect T-cells from patient

Genetic modification with viral vector

Virus encoding TCR or CAR

Expansion of cells

Re-infusion to conditioned

patient

Gene engineered T cells for Therapy

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A brief history of the UoM CTU….

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2003 to 2006 MFEz & aCD19z PD & PQ

2007 to 2010 MFEz & aCD19z

Trials

1990 to 2003 UoM – Prof Robert Hawkins

Basic science immunotherapy of cancer

2011 CTU MHRA IMP & MS licence 1st patient treated Oct

2011 under MS special

Dec 2008 to Feb 2010

CTU inception to opening

• Continue existing Gene therapy T Cell trials

• Develop novel cell therapy treatments for cancer and other diseases

• Must be effective, flexible and safe systems of cell engineering 5 products simultaneously

• Develop and train staff

• MHRA IMP & MS specials licences

Cellular Therapeutic Unit - Objectives

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• Simple and effective Quality management system

• Modular processing methods that can be used across products

• Move away from classical clean rooms

• Isolator technology controlled aseptic environment

• Protects patients cells from infection or contamination

• Allows rapid decontamination with vaporised hydrogen peroxide without chemical residues

• Allows multi product processing

Cellular Therapeutics Unit (CTU)

Cellular Therapeutics Unit - Trials

• CAR - A Phase I Study of CD19 Specific T cells in CD19 Positive Malignancy • Up to 20 patients – open to recruitment 6/20 treated response in 3/6

• TIL - METILDA - Randomised Phase II Study in Metastatic Melanoma to

Evaluate the Efficacy of Adoptive Cellular Therapy with Tumour Infiltrating Lymphocytes (TIL) and Interleukin-2 Dose Assessment

• Up to 90 patients – Opened March 2014 with the EU FP7 ATTACK - consortium • TCR - NY-ESO-1/LAGE-1 peptide expressed in a subset of tumours:

[1] ATTACK Trial 1 - Oesophagogastric cancer – single arm • Up to 28 patients across – Opened Sep 2014

[2] ATTACK Trial 2 - Metastatic melanoma Standard vs Optimised manufacturing

• Up to 42 patients – Planned to open Q3 2015

-14 -13 -12 -11 -10 -9 -8 -7 -6 -5 -4 -3 -2 -1 0 1 2 3 4 5 6 7

Mon Tue Wed Thu Fri Sat Sun Mon Tue Wed Thu Fri Sat Sun Mon Tue Wed Thu Fri Sat Mon Sun

Cell culture & expansion

Patient Conditioning prior

to infusion

Cytokines support

Cell infusion 5x109 - 5x1010 cells

Leukapheresis

Monitoring

In patient

Anti-CD19z CAR T cells

Functionality assay

Assess: Cell expansion Transduction efficiency

Transduction

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Days in culture

Adoptive T cell Therapy for Cancer: CAR immunotherapy – anti-CD19 T cells

-9 -8 -7 -6 -5 -4 -3 -2 -1 0 1 2 3 4 5 6 7 8

Mon Tue Wed Thu Fri Sat Sun Mon Tue Wed Thu Fri Sat Sun Mon Tue Wed Thu

T-cell processing & expansion

Cyclophosphamide 60mg/kg

Fludarabine 25mg/m2

Interleukin-2 Up to

12 doses

T-cell infusion ≥109 cells

Monitoring

Prophylaxis & GCSF

Tumour resection or Biopsy

Thaw & Initiate REP

TIL growth Patient eligibility report

Assess: Cell expansion Sterility Mycoplasma

Day -X Day -X +14 to 30 Day -17

TIL expansion either expand immediately

Or freeze

35±5 Days in culture

Natural T cell therapy - TIL

Adoptive T cell Therapy for Cancer: TIL Immunotherapy

CTU manufactured cell therapy for Metastatic Melanoma in MS special Aug 2012 Patient response 24 months • large mediastinal lymph node • Cerebellar metastasis

Adoptive T cell Therapy for Cancer: TIL immunotherapy

CTU manufactured cell therapy for Metastatic Melanoma in MS special Jul 2013 Patient response 6+ weeks • Sub cutaneous • Lung • Axillary disease

6 weeks after cell therapy Before cell therapy

Encouragingly all patients have had some tumour reduction so far…

Adoptive T cell Therapy for Cancer: TIL immunotherapy: 6 MS specials patients

Baseline PET scan January 2, 2013

Day 101 April 19, 2013. Normal heart

and tonsilar uptake

Synovial sarcoma Adoptive Therapy with: TCR Immunotherapy therapy - NY-ESO-1 Engineered T Cells.

CAR service history

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• September 2013 there were 111 protocols registered NIH - 104 of which targeted cancer, with more than 500 subjects dosed - 40 protocols address haematological malignancies & 34 targeting

CD19 • Key factors:

- The target antigen is expressed on the cell surface - Good clinical evidence in CD19+ tumors at multiple centres

No.

of C

AR

pro

toco

ls (N

IH)

Jacqueline Corrigan-Curay et al., Molecular Therapy, 22, 1564-1574

TIL immunotherapy

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• TIL • Isolation of TIL from tumour is complex and unpredictable • Unknown target difficult to prove specific functionality – trials using functional TIL did not correlate with efficacy • TIL therapy – Response in 40 to 50% of patients of which 20 to 35% CR • Current theory – highly mutated cancer > frequency of tumour specific T cells

Michael Lawrence et al., Nature, 2013, 499, 214–218

TCR immunotherapy

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• TCR • HLA dependant

- Patient must screen positive for HLA and antigen - High rate of screen fails i.e. HLA-A2 approx. 40% NY-ESO-1/LAGE-1 approx. 30-40% in Oesophago-Gastric cancer - 1 to 2 in every 10 screened • Evidence of activity in: - Melanoma – Response 9/18 (CR 4/18) - Synovial sarcoma – Response 10/16 (CR 0/16) - Multiple myeloma – Response 13/20 (CR N/A)

Toxicity associated with current T cell immunotherapy

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• Systemic conditioning chemotherapy – Systemic conditioning chemotherapy - lympho/neutropenia & sepsis – IL-2 – Pulmonary oedema/fevers/riggers

• Off target activity effects of cancer immunotherapy: • TCR associated toxicity seen with high-avidity TCRs targeting off

tumour epitope – MART-1, GP-100, CEA & MAGE-A3 • CAR associated off target toxicity – Carbonic anhydrase (CA) IX - Bile duct – CEA • TIL

– vitiligo & uveitis

• Active therapy in bulky tumours – Cytokine release syndrome – Tumour lysis syndrome

Challenges for the industry

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• Scale up to treat large numbers of patients Just in time manufacturing Shortage in manufacturing capability Labour intensive facilities and processes Secure supply of critical reagent & CoGs

• GMP vector for CAR & TCR therapy Shortage in manufacturing capability Cost barrier to do early phase trials

• Shortage of treatment centres Intensive chemotherapy High dose IL-2 management

ATTACK • Multicentre Phase II

– 2 trials with up to 70 patients – Trial 1 – Single arm

Oesophago/gastric cancer with n=28 patients

– Trial 2 - Melanoma 2arm n=42 patients

– Centralised Production • Manchester & Amsterdam

– Start Q3 2014

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7 -7 -6 -5 -4 -3 -2 -1 0 1 2 3 4 5 6

Patient Conditioning prior

to infusion

Cytokines support

Cell infusion 5x109 - 1x1011 cells

Frozen product

Monitoring

Scale up requires scheduled manufacturing & treatment

Manufacturing, Quality control

Cryopreservation Product release

Frozen starting material

In patient

NY-ESO-1 TCR T cells Phase II clinical trials – Oseophago/Gastic cancer & Melanoma

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Several weeks manufacture

10 Days Release testing

• To date less than 1,000 patients treated globally

• High value products & clinical results have lead to a movement from chemical drugs to biotherapeutics to ATMPs

• Huge increase in pharmaceutical interest

• Regulations have adapted to industry requirements (e.g.):

- ATMP regulations

- Multi product manufacturing

- Risk management principles (ICH Q9)

Future of manufacturing?.....in the CAR industry

2014

1914

Nov 10 CTU refurbishment

complete

Aug 09 2nd patient

treated

2005 2007 2006

2012

2008

2011 2010 2009

May 09 Major protocol amendment Submitted to

MHRA and GTAC

?

CD19 trial timeline – lack of experience as an industry

May 05 Application

to GTAC

June 05 GTAC

approval

Mar 06 Tegenero

Anti-CD28 trial

Nov 06 Application to MHRA

May 07 MHRA

approval May 07

GMP retrovirus available

Jan 09 1st patient

treated

Mar 10 Study suspended NBS lab closure CTU established

Jun 09 MHRA

And GTAC approval

Oct 07 GMP aCD19z process qualification complete

Mar 08 Christie R+D

approval

Oct 09 3rd patient

treated

Q4 10 – Q4 11 CTU aCD19z GMP process

Requalification & facility

validation Q4 11

CTU MHRA License approved

2011 2013 2012

2018

2014

2017 2016 2015

NYESO-1 T cells – better understanding

EU Grant

application

April 2012 EU approval

Q3/2012 CTL Process development

OG Trial Protocol

IMPD Feb/13

GMP process validation (enough virus for 1 run)

Nov 2013

OG Trial ETHICS MHRA

Approval Jun/13

Delay Availability

of virus

Delay only due to contractual agreements

OG Trial Trial

OPEN Sep/14

Acknowledgements Program Lead Robert Hawkins Cellular Therapeutics Unit Ryan Guest Natalia Kirilova Julie Duckworth - CTL Martine Thomas Fiona Baluwa - CTL GMP assays & GCLP Clinical Monitoring Group Dominic Rothwell Debbie Burt Experimental Cellular Therapy Group David Gilham Vicky Sheard Hannah Gornall Vania Baldan CTU Design & Implementation Ryan Guest Nikki Price - CTL Angela Osborne Clive Brooks Gavin Sutton

Clinical Cell Therapy Fiona Thistlethwaite Was Mansoor Kerry Dunn Michelle Davies Jenny Haughton Lorraine Turner Tracey Heslop Melanoma Group Paul Lorigan Jackie Hodgetts Christie Hospital Andrea Spencer-Shaw Rita Dowse Sponsor/Christie Clinical Trials Unit Ian Emerson Phil Barley Angela Ball GMP TIL manufacturing - Harmonisation Team Bianca Heemskerk – NKI-AVL Joost vd Berg - NKI-AVL Marco Donia – CCIT/Herlev Hospital Ryan Guest – University of Manchester/Christie

Research Management Nicola Price - CTL Helena Kondryn TIL Advisors Mark Dudley Michal Besser Surgeons Aali Sheen Ajith Siriwardena Mark Jones Piotr Krysiak David Sherlock Deemesh Oudit Gary Ross Vijay Ramani