Ensuring Quality Management of PK Samples from Collection to Archival and Selecting the Right Biorepository for Your Nee

1. Ensuring Quality Management of PK Samples from Collection to Archival and Selecting the Right Biorepository for Your Needs Gina McMullenSr. PK Research AssociateISIS Pharmaceuticals, Inc.

2. Objectives: • Provide an overview of PK sample collection, processing, analysis, data reporting, and eventual archival. • Identify considerations for protocol planning & Bioanalytical Lab selection to ensure integrity & accurate data reporting. • Evaluate key aspects when choosing a commercial Biorepository for long term storage (of PK samples), in order to maintain integrity for potential future use, and ultimately benefit your drug development program in the long run.

3. Example of First Order Kinetics What is PK? • Pharmacokinetics (PK) is essentially the study of what the body does to a drug once it’s been administered by a chosen route (i.e. Topical, Enteral, or Parenteral). • PK samples are collected at precise time points (per protocol) in a specific type of biological matrix (typically plasma) & subsequently analyzed for drug concentrations. • Bioanalytical methods used to analyze these samples should be selective and sensitive, in order to construct an accurate concentration-time profile for your drug.

4. The Importance of PK Samples • Pharmacokinetic (PK) samples play an integral role in the drug discovery & development process. • PK concentration determinations not only allow for accurate characterization of the drug, but also assist in establishing the most effective dose regimen for the intended patient population. • Ensuring these samples are properly collected, processed, analyzed, and eventually archived, is critical to the success of your drug.

5. General Overview of Drug Development Timelines Studies published in 2003 indicate the cost to bring a new drug to market is around $800 million.[1,2] More recent studies estimate that cost to be as high as $2,000 million, depending upon the therapy or the company developing it. [3]

6. Preclinical IND Enabling Toxicology Studies Phase 1, First in Human Studies Phase 2A/2B Studies Phase 3 Studies PK Component of Drug Development Timelines Up to1000 PK samples generated for analysis in a single IND enabling Toxicology study (nonhuman primate study) Up to 1500 PK samples generated for analysis in a single Phase 1, FIH, SAD/MAD study design (35-40 collected per patient) Around 500-1000 PK samples generated from a single Phase 2A/2B study depending upon number of patients enrolled (25-30collected per patient) 1000+ PK samples generated from a Phase 3 study, but only around 5-10 collected per patient (increased # patients enrolled) Time to Market

7. General Considerations The Importance of proper PK sample collection, processing, reporting, and eventual archival

8. Pack sufficient dry ice in shipping container to ensure samples won’t thaw in event of delays. Two transfer tubes labeled identically: 1) Primary Aliquot for initial analysis 2) Duplicate Aliquot as a backup Purple topped EDTA collection tube Ensure frozen immediately at appropriate temperature (i.e. -80oC) Proper identification at PK Lab is essential to accurate data reporting i.e. 1500g for 15 min @ 4oC Overview of PK Sample Collection to Data Reporting Use validated methods for PK analysis under GLP compliance to ensure quality final data. Centrifuge Freeze Sample Log-In Ship to PK Lab for Analysis Analyze samples and report data Split into 2 equal aliquots Collect Sample

9. PK Life Cycle Study Considerations in order to ensure sample viability for future use Protocol Planning for Sample Collection • Select optimum time points for drug characterization and identify in the protocol • Communicate sample collection & processing expectations • Type of collection tube (i.e. K2EDTA) • Centrifugation speed, time, & temperature • Amount to be apportioned to separate transfer tubes • Specify information contained on tube labels • Importance of assigning primary & duplicate aliquots for sample analysis availability • Temperature for storage PK Analysis • Ensure bioanalytical method has been validated prior to analysis • Verify samples clearly identified at time of collection and are correctly reported in final data set Eventual Sample Archival • Proper handling & management will allow samples to be utilized for additional analyses in the future if needed

10. Important Roles in Clinical PK Studies Clinical Site • Ensure protocol is followed and samples are properly identified at time of collection. Central Lab • Preparation of PK Kits for Clinical site, including collection tubes & labels. • Collate samples from multiple Clinical sites for shipment to designated PK Lab: • Initial sample identifier reconciliation. • Potential shipment directly to Biorepository. PK Lab • Maintain information provided by Central Lab through analysis to preserve integrity of samples and ensure quality concentration data. • Additional sample identifier reconciliation if needed. • Subsequent shipment to BioRepository upon completion of analysis and final data submission. BioRepository • Retain value of samples by ensuring proper storage for potential future use.

11. Bioanalytical Considerations during Protocol Planning • Matrix Selection • Clinical: typically plasma or urine, occasionally tissue biopsies • Preclinical: typically tissue, plasma, and urine for initial IND enabling studies • Species selection (i.e. rodent, non-human primate, canine, etc.) • Select appropriate type of assay to analyze PK samples in specified matrix • LC/MS • CE • Ligand Binding Assay (ELISA)

12. Additional Considerations • Select appropriate bioanalytical facility for future PK sample analysis • Ensure viable assay for concentration determination and efficient method transfer • Establish sample collection, stability & storage requirements • Anticoagulant to support analysis • Detailed Processing instructions • Transfer tube type (i.e. polypropylene vs. polystyrene, etc) • Labels that will adhere to these tubes during freezing process • Storage temperature & light sensitivity considerations to prevent degradation • Limit number of freeze/thaw cycles to maintain sample viability

13. PK Study Considerations • For Clinical Trials, identify suitable central lab to ensure adequate supervision of clinical sites, sample management, & subsequent shipment to PK lab for analysis • Preparation of sample collection kits, forms, labels, supplies, etc. • For Preclinical Studies, ensure chosen In-Life facility can employ proper collection/processing/shipping/ and documentation technique. • Proper information to be included on sample label (based on protocol) to ensure accurate identification for future use • Requirements typically include the following: • Protocol • Accession # barcode • Patient/Animal # • Collection Date & Time (and/or Study Day & Time point per the protocol) • Matrix type (plasma, urine, etc.) • Aliquot assignment (primary vs. duplicate)

14. Investigator Information Barcode Label Visit information Patient information and sampling information Airway Bill Number and comments Number of specimens collected/ name, signature and date Send the white copy of the form with ambient specimens to Central Lab. Send the yellow copy of the form with frozen specimens to Central Lab. Retain the pink copy at clinical site. Documentation of Proper PK Sample Collection in a Clinical Trial Complete and check all the information on the Laboratory Requisition Form IMPORTANT Ensure the Laboratory Requisition Form is complete and legible. If an error is made, strike-through the wrong information without obscuring the original entry, correct it, sign and date the change. Once completed, this form must be faxed to the Central Lab the day of sample collection. This is for tracking purposes. Please use the forms as follows:

15. Shipment Details • Current address of PK facility plus contact information for individual who will be responsible for sample receipt • Especially important when procuring specialized shipping permits. • Detailed shipping manifest • Preferably electronic to be populated with electronic data • Temperature of sample shipment (wet vs. dry ice) • Estimated shipping schedule (i.e. batching samples based on collection time points) • Preferred courier for shipping (company who will ensure temperature is maintained throughout process & will re-fill ice if shipment is delayed) • Tracking # allowing you to check status of shipment on-line

16. Sample Log-In • Upon arrival at PK Lab • Notification to sponsor of safe receipt • Sample log-in (discrepancy reconciliation between tube label & manifest) • LIMS (Laboratory Information Management System) for internal sample tracking at PK lab

17. Sample Analysis & Data Reporting • Use validated method for analysis • Proper handling of samples during analysis can prevent contamination • If insufficient volume in primary aliquot use duplicate (ensure accurate identification and reporting). • Ensure sample identifier information is accurately reported in the final data (i.e. tracking system) • Can also be used as a manifest for shipping to selected off-site Biorepository • QC/QA review of data and finalization of report • Watch for anomalous concentration values which might otherwise be explained by incorrectly reported sample identifiers or contamination.

18. General Considerations Choosing the Right Biorepository for long term storage needs

19. Key Traits to consider: • Interaction • Identification • Inventory • Invoicing • Integrity

20. Interaction (ensuring quality communication with your chosen Biorepository): • Communicate with Biorepository to ensure they can accommodate proper storage requirements: • Optimal storage temperature (i.e. < -80oC) • Light sensitivity concerns • Length of time for storage requirements: • Minimum amount of time for which samples need to be stored? • Maximum amount of time they are allowed to be stored? • What is your established Long Term Matrix Stability for this compound? How does that compare with industry guidance?

21. Interaction (cont.) • Shipping considerations to Biorepository • Ensure constant temperature in shipment • Clearly labeled sample tubes • Accurate shipping manifest, (preferably electronic), which can easily be linked back to information on tube label • Have Biorepository coordinate with Analytical facility to ensure safe & timely shipment of your PK samples

22. Identification (being able to properly identify a specific sample if/when needed): • Sample identifiers - how will facility be able to locate samples if needed in the future? • Do they affix a specific label upon arrival at their facility during initial log-in of the samples? • Do they have an in-house tracking system? If so, what type? (Paper, electronic, etc.) • Policies on discrepancy resolution if samples arrive and labels don’t match with the shipping manifest. • Additional cost to sponsor? • Utilize PK lab to prepare shipping documents and QC labels prior to sending for long term storage?

23. Inventory (knowing what you’re storing and how much): • Double check policies re. sample inventory: • Can sample information be accessed on-line? • Does this feature result in additional charges (i.e. only certain individuals can access )? • Will hard copy of inventory be sent? If so, how often? Monthly, quarterly, annually? • Determine in advance how long samples should be retained (per industry guidelines & established LTS): • Plan accordingly to have them disposed of at designated time points to ensure room for new samples.

24. $ Invoicing (budgeting accordingly for your Biorepository needs): • Know what you’re being charged for and how often: • Per sample cost vs. entire shelf/freezer? • Monthly bill vs. quarterly or annually? • Hidden fees? • Cost to locate/ship samples if needed? • Cost to destroy samples once established LTS has been reached? • In what capacity will you be utilizing your Biorepository? • Batch shipping samples from ongoing studies for future analysis vs. long term storage once studies have completed?

25. Integrity (acknowledging the value of these samples and ensuring the appropriate measures are in place to retain their value): • Samples are irreplaceable and loss or damage to them would result in delays to your drug development program. • To ensure sample integrity, your chosen facility should have measures in place to account for the following: • Additional freezers for moving/ transitioning samples • Back up generators for power failure • In case of natural disaster are steps in place for additional generators/fuel/ manpower? • Sufficient storage space to accommodate new freezers for future samples? • Multiple locations to accommodate different shipping sites

26. Take Home Message • Maintaining sample integrity for potential future can be achieved by employing proper planning and diligence in the following areas: • Sample Collection/Processing • Sample Analysis • Long Term Storage …which can ultimately add value to your drug development program!

27. References • DiMasi J. "The value of improving the productivity of the drug development process: faster times and better decisions". Pharmacoeconomics 20 Suppl 3: 1–10. PMID 12457421. • DiMasi J, Hansen R, Grabowski H (2003). "The price of innovation: new estimates of drug development costs". J Health Econ 22 (2): 151–85. doi:10.1016/S0167-6296(02)00126-1. PMID 12606142. • Adams C, Brantner V. "Estimating the cost of new drug development: is it really 802 million dollars?". Health Aff (Millwood) 25 (2): 420–8. oi:10.1377/hlthaff.25.2.420. PMID 16522582.

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