Development of a Point of Care preparation device for Cell Therapies

Martin Smith, Closed Cell Systems13th AMC Technical meeting

Overview

• The Healthcare Challenge for Cellular Therapies

• Cell Preparation (Cprep) system– Concept & early work (Phase1)– Development & Commercialisation (Phase 2)

• Closed Cell Systems

Cell Therapy Healthcare challenge

• Cell therapies offer cures for patients and reduced through-life costs.

• Cell therapies are expensive(> £20k typically).

• A shelf life of only a few hours is a huge logistical and administration constraint.

• Freezing removes the shelf life constraint, but often requires cryoprotectant removal at the PoC.

Scale-out challenge; processing at PoC?

• Clinical Research Facility with licensed GMP clean room.

• Cryoprotectant removal by thawing, dilution, centrifugation in classified area/isolator

Overcoming the scale-out challenge

• Cryoprotectant removal is a manufacturing step within the manufacturer’s license.

• CPrep automates this process in a closed environment suitable for use in an unclassified area, and includes the necessary patient tracking and release information.

• Its benefits include, lower cost (50% cheaper), lower risk, remote release.

Cprep. Concept & early work

• 2012-2015 Innovate Project 101098– UCL– eXmoor pharma– LONZA– Amercare

Side View – Wash operation

Cell Suspension

outlet

Bottom flow inlet

The Design:

1. The membrane filter is used as a cell retention device;2. Cell settling is a part of the design (for Fibroblasts)

Novel cell chamber development

3. Water displacement (i.e. volume reduction) by air pressure;

membrane

membrane

Side View – Concentration

inlet

outlet

To syringe

Air bubbleThe Design:

Novel cell chamber development

3. Water displacement (i.e. volume reduction) by air pressure;

Manual Test Rig at UCL

Heater Block

Cell Chamber

Aspirating needle (Vial)

Syringe Pump

Cprep Processor V1

• Fully Automated• Aseptic vial dock and closed

process• Single Use Disposable;

– One touch, error free placement

• Rigid Frame Disposable comprises;– excipient buffer, cell chamber,

product syringe, waste container

• Bag Expressers drive fluid flow through vial and cell chamber

Cprep Operation

• Starting with the device, disposable and vial• Confirm patient, vial and paperwork etc• Switch on device, self check, log on• lock in disposable• remove vial from shipper and install• start sequence

– controlled thaw & priming– automatically dock vial to disposable– Priming and cell wash into chamber– wash sequence begins– air concentrations and fills syringe at target volume

• Print BMR, Release, Administer.

Regulatory considerations

• Processor will be regulated as GMP equipment. • The processor forms part of the Cell Therapy Manufacturers (CTM) process.

• Disposable is a GMP consumable• Disposable will be manufactured, assembled, filled, wrapped, labelled and

sterilised by a reputable single use supplier.

• Cell therapies will be released as follows • CTM part certification before dispatch.• Point Of Care inspects and retains both logistics and CPrep records.• CPrep processing by trained operators.• Product released locally by suitable person (after review of records).• Cell Therapy administered• CTM completes final certification (after administration)• CTM holds all records as part of the BMR including the CPrep element

End of Concept Phase 1

• Test rig built and many operating configurations / geometries tested.• Viable cell recovery similar to conventional centrifugation control. • “Best” arrangement patented and built into demonstrator.• Heads of terms agreed and license drafted from phase 1 collaboration to

CCS.

N ≥ 3 for each data point

1. DMSO removal

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20

40

60

80

100

Run 1 Run 2 Run 3 Run 4Input viability (%)Output viability (%)

3. Viability

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0.2

0.4

0.6

0.8

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1.2

Run 1 Run 2 Run 3 Run 4Input Volume (mL) Ouput Volume (mL)

2. Volume reduction

Commercialisation Phase 2

Outcome:•Production version + supply chain.•Trials demonstrating

– clinical utility & effectiveness. •Document package for regulators.•Market testing and order pipeline.

WP1 Test demonstratorWP2 Innovate improvementsWP3 Processors and disposables.WP4 Test production equipment.WP5 Process development on CT1.

JV: eXmoor pharma & Amercare

Cprep V1 Bag Expresser issues

Bag Expresser concept• Significant spring force

required to compress bags– (high surface area)

• Force varies as bag empties• Control algorithm too complex

• Cprep able to provide equivalent motor force but not without unmounting the disposable

Open heart surgery

3D printing…to the rescue

Reverse engineer peristaltic pump

Scale to fit existing motor drive

3D print pump and housing

1st iteration

2nd iteration

3rd iteration

Cprep V2 system development

Cprep V1.5 Cprep V23D CAD

Single Use Disposable

3D Printed Prototype Process Diagram-Wash step

Process Development Laboratory

• Establish high quality and consistent cell banks– 30 x1mL vials 10^7 MRC-5 Lung Fibroblasts– 300x1mL vials 10^7 umbilical cord MSCs– 300x1mL vials 10^7 T-cells (Jurkat)– 1mL ±50uL, 10^7cells, ±5% per vial

• Dedicated scientist & lab space– 2xCPrep runs per day, 4 days per week.– QC (counts, viability, growth at t=24h– Qualified HPLC method for DMSO

• Flexible, cost effective, collaborative partner– Assistance with qualifying aseptic dock– Host site for client and regulatory demonstrations– Willing to journey with us

Process Development Laboratory

Fibroblast Cell Banking

• Establishment of high quality and consistent cell banks:– 30x1mL Fibroblasts at 1*10^7/vial

Hand filling of low vial numbers

Process Capability Analysis

Baseline Centrifuge Controls

• Cprep automated thaw– Manual aspiration– 1:10 dilution– centrifugation 5min 300g– Resuspension in 0.5mL PBS

Cell Type Initial [DMSO]

Final[DMSO]

Yield post centrifugation

Fibroblasts (n=5) 4.6±0.02% 0.12+0.03% 89±4%

T cells (n=5) 4.6±0.01% 0.37±0.1% 74±5%

uMSCs (n=3) 4.3±0.03% 0.13±0.05% 85±2%

HPLC method for [DMSO]

• Method Transfer• Literature & HPLC vendor

review• Adaptation• Qualification• Validation• Maintenance• <7-10d turnaround

– Standard curve, 0.025 to 5%– LO Quantitation =0.027%– LO Detection =0.008%.

Cprep experimentation

Computational Fluid Dynamics

• Modelled Flow behaviour as function chamber geometry

• Fibroblasts modelled during mid-wash prior to partial settling on chamber floor

Cell Mass Balancing.

• Chamber A3, 3x10^6

• Chamber A3, 8x10^6

Cprep experimentation & CFD-Phase 2

Membrane Type

Membrane Pore Size

Chamber Design & Geometry

Inlet diameter

Wash Flowrate & Volume

Backpulsing

Concentration, rates & amounts

Chamber Materialsof Construction

Chamber surfaceroughness

Needle diameter & orientation

Needle height within vial

Different cell types & viability

Cell concentration

Syringe fill rates

System & Chamber prime

• 235 cell based runs• 4 cell types• 6 chamber

prototypes• 4 processor

designs

Working with Prototype materials…..

Typical Cprep results

8-10*10^6 cells/vial

A3 chamber Yield (%) [DMSO] (%)

Cell/Condition 30mL 60mL 30mL 60mL

Fibroblasts 80% 73% 0.56% 0.25%

T cells* 60% 10% 0.68% 0.18%

uMSCs 86% 80% 0.70% 0.37%

• 0.4um tracked etched membrane, (*0.2um)• 1mL/min• PBS wash buffer• Ambient processing• Prototype acrylic chamber

Aseptic Dock and frozen vial detection

• Control software detects frozen vial

• Air tight Septum closes aseptic dock

Commercialisation update

• Processor V4 • Control system qualification

• Regulatory Acceptance

Q3 15 Q4 15 Q1 16 Q2 16 Q3 16 Q4 16 Q1 17 Q2 17 Q3 17

Test Demonstrator

MHRA Review

Client Demonstrations & Evaluations

Confirm First Client

Improve Demonstrator

Production Engineer

Production Supply Chain

Test Production Device

Preparation for 1st Clinical Trial

Commercialisation Timeline

• eXmoor will grow PD capability• Amercare will supply processors• CCS will sell processors and

disposables• CPrep is the first product of an

expanding portfolio for CCS.

• Amercare and eXmoor jointly own Closed Cell Systems Ltd (CCS) to:• Combine conventional process development, with• engineering new equipment, and then• selling the resulting processors and disposables

Closed Cell Systems Ltd.

Summary

• The technical development of new cell therapy equipment in a heavily regulated environment is difficult.

• Balancing process development with processor / controls / disposable development

• This machine works – it achieves its targets• This machine helps to make cryopreservation of very

expensive, short shelf life products possible• We are ready to introduce this machine into your cell therapy

manufacturing process• We are also interested in helping with the solution of other

atmp manufacturing challenges.

Acknowledgements

• NHSBT– Charlotte Cox– Linda Motz– Richard Forsey– Martin Guttridge– Paul Lloyd Evans– Antony Wright– Jon Smythe– Teresina Pinnington

• Amercare (& associates)– Graham Wilson– Lee Harrington– James Aitken– Oliver Sedlacek

• eXmoor pharma– John Vincent– Angela Osborne– Rob Scott

Collaborations and Evaluations

info@closedcellsystems.com +44 (0)1398 371 463