Lucinda H. Cohen, Director, DMPK NJ Bioanalytical Group Merck Research Laboratories

1. Let the Robot Do the Work: A Global Automation Strategy in Drug Metabolism & PharmacokineticsHamilton Mid-Atlantic User Group MeetingDecember 9, 2010 Lucinda H. Cohen, Director, DMPK NJ Bioanalytical Group Merck Research Laboratories

2. Outline • Setting the stage – the case for change • Baseline assumptions in designing an automation program • Implementation • The next generation • Lessons learned

3. In the beginning....

4. On NOT Reinventing the wheel

5. Talents Skill sets Training Relationships Culture People Equipment Processes Inputs Outputs Customers Business needs Metrics

6. Past/Current State Study PI Sample Preparation Mass Spectrometry Data Analysis and Reporting Variable Results, Timelines and Costs Lab 1 Lab 2 Lab 3

7. Current State of Preclinical Industrialization (Year 4 of 5) Study Request System LIMS Automated Data Analysis Study PI Standardized Reporting Project Teams Qualitative UHPLC-HRMS Automated Sample Preparation Shorter Timelines and Lower Cost Automated CompoundTuning Quantitative UHPLC-MS

8. A Global Automation Strategy One site (RY) collaborates with Hamilton Co. to develop highest usage method Second site (MF) beta-tests program and begins implementation Automation leader travels to remaining sites (WP, BO) to facilitate full implementation Duplication of effort avoided, consistent harmonized process utilized Working group met monthly in Year 1 to prioritize and develop new automation functionalities

9. “Begin with the end in mind.” – Stephen Covey • An automated sample handling and extraction routine to support routine rat plasma PK • >90% of rat PK experiments will be prepared using Hamilton after implementation • Focus is on large sites and Merck Frosst • Development in collaboration with Hamilton and originating from Rahway

10. Rahway Biomek 2000,3000, FX Hamilton STAR Boston Hamilton STAR+ IRBM Hamilton STAR+ Banyu --- West Point Preclinical Biomek 3000, FX units Tecan units Hamilton STAR Clinical: PE Janus Packard Multi-probe Global Technologies – now defunct PGP assay - Tecan Automation instrumentation – site capabilities at onset

11. Data collection - survey

12. Elements of Core Program • Preparation of standards, QCs, blanks, samples and diluted samples • Addition of internal standard via ACN crash solution • Plate shaking • Tip sparing algorithm • Initial program for plasma, quickly expanded to mouse blood PK (exact volumes of blood in citrate buffer)

13. Design flexibility • User inputs study design: text delimited file • Hamilton interface for manual parameter input: • stock solution starting concentration • Text file name

14. Controlled vs. Flexible Parameters Controlled • 50 uL plasma aliquot • EDTA plasma • ACN as crash solution • Plate shake time (5 min) • Final plate format • Plasma blank Flexible • Standard curve range and concentrations • # standards • # and concentration of QCs • # samples • Sample dilutions

15. The Quality Controls Dilemma

16. What were our options? • Gold standard is <5% organic in any standard/QC • Pfizer Ann Arbor program’s method of initial organic addition with subsequent plasma dilutions led to issues with low QCs • Ultimate solution of <5% organic with dilution strategy to generate acceptable QCs would have required significant programming and method changes • Issues with either Hamilton or West Point preclinical BA method would not be discovered until GLP validation or Clinical BA method transfer

17. Implementation – the rubber meets the road for PK • Merck Frosst actually did beta-testing and fully leveraged Hamilton immediately • Initial rollout in RY used Open Access concept, was centralized with a contractor • West Point uses in centralized manner, rotates scientists in/out of Hamilton “workstation” • Boston uses contractors, hybrid approach Bottom line impact: Global improvement in bioanalytical cycle times, routinely analyzing >80% studies in 4 business days or less

18. Long Term Initiatives • Other matrix types • other species’ plasma • mouse blood • rodent tissues • Submission of multiple batches: current program has 4 plate capacity • Error response and reporting • Dynamic queueing: towards a 24/7 operation? • Additional deck functionality: centrifuge, sample plate hotels, tip stacking? • Sample pooling

19. PK Scientists PD Scientists Previous Workflow Aria/Analyst Analyst User: Creates/modifies sequence Creates new .DAM file Phact Watson Create R number Generate sequence Operator Hamilton Hamilton User: Reference R number Get sequence file for chemists Insure correct consumables on deck

20. PK Scientists PD Scientists Current Workflow Phact Watson Hamilton DB Job List Protocols Plate Map Aria/Analyst Create R number Generate sequence Analyst User: Barcode is used to compile new sequence and dam file Hamilton User: Loads 3 different matrix and consumables Based on barcode, robot will assign correct protocols Hamilton

21. Phact Software HTTP User Workstations Web App ( HTTP ) HTTP Hamilton Workstation Technician Hamilton Robot Architecture Diagram Main Server Phact DB Mongrel Rails Service SQL SQL MySQL DB Static Files Apache Web Server Worklist File

22. Web interface

23. Job data can be either entered on the web or imported from XML files • Job data can also be exported to XML files XML with Plate Data Enter data on the web

24. After data are entered, job is saved and can be visualized on the web interface • Data can be modified quickly through the web interface as needed Job data can be saved into another copy as template

25. Validation performed after data entry • Labels enable lab operators to identify plates and tubes • Detailed information is accessible through the barcode PROJECT DP L-123456 Rat Wistar L-123456 PROJ DP

26. Dosing solutions and samples are transferred between geographically distant departments at Merck An online tracking tool was developed to track sample life cycle and delivery Sample Tracking & Workflow Management

27. Key Advantages • Hamilton technician scans the plate barcode(s), inputs consumables info. Data capture and parsing on the Hamilton system is completely electronic. • Barcode system facilitates easy and correct labeling of plates, enables plate tracking and metrics collection • Elimination of paper forms greatly reduces the opportunity for errors • Web application provides a user-friendly interface to enter job information with data validation, and visualize data vs. physical samples • Template functionality allows users to quickly clone standard jobs and plates, reducing redundant data entry

28. Outcomes • A method has been developed to centralize and simplify the data entry process and to automate worklist generation for the Hamilton Star robotics platform • The web application centralizes data collection regarding sample plates and job parameters from users • The barcode from the sample plate is the only piece of job information required from the robot operator • The server generates sequence files in the same format as before, allowing seamless integration with the current software • The use of a database to track job information allows robot usage metrics generation

29. Lessons Learned • Liquid handling capabilities of Hamilton system have been assessed and continue to perform per specifications. • Monitored air displacement (MAD) is key to Hamilton functionality • Process has been (non-GLP) “validated” and implemented. • Original goal of 80% in vivo studies prepared using Hamilton was exceeded. • Implementation of Hamilton approach allowed more work with fewer people • Goal is to free up time from routine/mundane work to focus on more cerebral, “value-added” research

30. Lessons Learned, cont’d. • Standardization is a laborious process. Define a time period for debate, end it and implement. Once the new process is in place, people generally accept the new paradigm and move on to different complaints. • Automation can drive much more standardization than expected. • Change will take much longer than you think it should. • Few people lose their aversion to change, but many improve their adaptability (kinetics!) • Recognizing and rewarding the change agents will push the process forward.

31. Rahway Bernard Choi Sharon Tong Mike Wolff JoAnna DiVincenzo Kyle Lady Rong Li West Point Deb McLoughlin Ken Wilson Ken Anderson Rick King Merck Frosst Robert Forget Laird Trimble Kevin Bateman Banyu Norihiro Takenaga Takuro Hasegawa Boston Andreas Harsch Todd Stawicki IRBM Enzo Pucci (Boston) Acknowledgments Hamilton Marek Rybka Roberta Ross Steve Corron Brandon Houchins Merck Executive Sponsorship: Lisa Shipley Tom Baillie Dennis Dean Jane Harrelson Rick Morrison Greg Winchill