Merck Microgravity Biologics Crystallization Experiments

Paul Reichert Structural Sciences October 15, 2014

CASIS Connection

• Solicit proposal for protein crystallization experiments

− identified as processes which could benefit from microgravity

• Proposal emphasis; benefit to human health – Cost: payment in kind – Minimal astronaut intervention – Use flight certified hardware – Ground control – Legal agreement emphasis; Publish rather than IP

• Peer reviewed selection of experiments

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Pharmaceutical Applications for Crystalline Biologics

Area Goal Application

Drug Discovery

Manufacturing

Product Development

– Structure determination – Structure based drug design

– Purification – Storage/stability

– Controlled Release – High Concentration formulation – SC and Pulmonary Delivery

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Why Microgravity? • Microgravity Enhanced Crystallization Effects:

– Crystallization Physics • Reduce sedimentation

• Minimize convection currents

• Reduce molecular diffusion rates

– Crystal effects • Larger

• Higher order

• More pure

• More uniform suspensions

• Learning experience (Entrepreneurs, hardware, methodology, researchers, etc.)

• COOOL!

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www.nasa.gov

Merck’s Microgravity Experience: 11 Space Shuttle Missions as Secondary Payloads (1993-2003)

Application Flight Results:

Flight vs. Ground

Drug Discovery STS- 51 Larger crystals

Drug Delivery: Controlled Release

STS-70,101 More uniform suspensions

(animal studies)/better dissolution properties

Drug Discovery STS-107* Structural information

Mixed Results STS-51,63,68,

71, 73,79,83 and 89

Poor quality crystals (multiple scrubs, crew activity,

hardware issues, etc.)

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* Final fatal mission of Columbia, February 2003

MRL’s Proposal Crystallization studies of monoclonal antibodies being investigated for multiple human diseases

Mab2

I. Microgravity growth of single crystals for structure determination (SpaceX-3, April 2014)

II. Microgravity growth of crystalline suspensions for multiple pharmaceutical applications; drug delivery, purification and storage of bulk active (SpaceX-6 planned for mid-year 2015)

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Mab1

MERCK, CASIS and NASA Scientists At Work in SSPF Lab Preparing Samples

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Loading Growth cells (PCF) NASA, CASIS and Merck loading team

Secured Labs

Protein Crystallization Hardware and Temperature Controlled Storage Units

8 PCG Hardware with 27 C GEL Packs

PCG Hardware in Flight Stowage Bags Ground Samples Storage (Control)

Flight samples in the stowage bag

Liftoff of SpaceX-3 Falcon 9 from Cape Canaveral AFS, April 18, 2014 3:35 PM

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Biologics Crystallization aboard ISS: Activation & Deactivation (Apr 21 to May 16)

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NASA Astronaut Steve Swanson

Photo Microscopy (Mab1)

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Ground (A5)

Flight (A3)

Plans for Future Flights (SPX-6) ~ 2Q’2015

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Space X-5 (November 2014) • Samples: Mab2 and Mab3 • Conditions:

4o C or 22o C …. ramp to 30o C in MERLIN incubator • Hardware:

High Density Protein Crystallization Facility (HD-PCF) Hand Held Protein Crystallization Facility ( HH-PCF)

Acknowledgements

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SpaceX-3 Experiment: Paul Reichert1, Giovanna Scapin1, Winifred Prosise1, Mark McCoy1, Steven Geng2, Suzanne Mercorelli3, Marianne Hayes4, Chakravathy Narasimhan5, April Spinale6, Ray Polniak7, Xiaoyu Yang8, Alexandre Ambrogelly8, Jeff Zhang9 and Ramesh Kashi10

Sr. Management: Dave Pollard, Mohammed Shameem, Steve Farrand (Bioprocess) Corey Strickland, Frank Brown and Chris Hill (Global Chemistry)

1Structural Chemistry - Structural Sciences, 2Rensselaer Polytechnic Institute, 3Biologics and Vaccines Formulation -Testing, 4Biologics and Vaccines Formulation – Potency and Functional Characterization, 5 Biologics and Vaccines Formulation – Design and Material Compatibility, 6CASIS Integration, 7NASA Quality Assurance, 8 Biologics and Vaccines Formulation-Process Characterization, 9Molecular Discovery–Protein Purification, and 10Biologics and Vaccines Formulation – Bioprocess Technology & Expression

Space Shuttle Experiments: Paul Reichert, Gerald Hammond, Peter Orth, Alan Hruza, Susan Cannon-Carlson, Wasyl Sydor, Suzanne Mercorelli, Fred Ward, Roland Mengisen, Yan-Hui Liu, T.L. Nagabhushan, B. Twarowska, U. Mirza, S. Wang and Hung V. Le UAB/CBSE Staff Marianna Long, Karen Moore, Vickie King Johnson, Dan Connors, David Ray, Lisa Smith, Charles Bugg and Larry DeLucas (This work was funded in part by NASA Cooperative Agreement NCC8-246)