Part of the MaRS Best Practices Series. Speaker: Valentia Lee-Brotherton, PhD, Ashuren. This session, led by seasoned industry experts, will explore how to effectively set up your pre-clinical POC studies, address pre-clinical safety requirements and issues, and give you an overview of the manufacturing standards required for Phase I studies. More information: http://www.marsdd.com/Events/Event-Calendar/Best-Practices-Series/ind-05132008.html

1. Accelerating Development
Preclinical Development
Planning for Emerging Pharma
and Biotech Firms
Valentia Lee-Brotherton, PhD
May 13, 2008
MaRS Workshop: From Benchtop to IND

2. Objective of Presentation
• To provide some points to consider when planning
and conducting a preclinical development program
to enable First-in-Human (FIH) studies
• Assumptions:
– Candidate selected (i.e. screening completed)
– Money in the bank (~3-4 $MM)
– Sufficient test article
– New Chemical Entity (not generic or reformulation)

3. Typical Timelines for Non-Clinical Testing
Programs
In-House Discovery
and Candidate Selection
Chemistry, Stability, (Bio) Analytical Development
Nonclinical Toxicology, Pharmacokinetics,
Safety Pharmacology
Preparation of Regulatory Documentation
Phase I Clinical Trial(s)
0 1 2 3 4 5 6 7 8 9 10 11 months

4. Challenges to the Emerging Biotech Company
• Lack of Resources • Investor Pressure
– $ – Meeting (realistic)
timelines in the face of
– Experience in product
issues (e.g., poor
development
candidate selection)
• Clearly Defined Vision
• Managing multiple
– Lack of clinical plan service providers
• Manufacturing/CMC – Contract laboratories
Issues (preclinical, analytical)
– Lack of test article – Contract manufacturer
(supply of test article)
– Formulation, scale-up
issues – Clinical CROs
– Consultants (clinical,
nonclinical, manufacturing,
regulatory affairs,
biostatistics, etc.)

5. The IND Application: A Significant Effort
• What's Needed?:
– General Investigational Plan (clinical development plans overall)
– Investigator’s Brochure (“IB”)
– Proposed Clinical Trial Protocol(s) and Investigator Information
– Chemistry, Manufacturing, and Controls Information
– Pharmacology and Toxicology Information
– Previous Human Experience
• Preparation Time
– 3-4 people, 1 month
• How Big?
– Typically 10-15 volumes (2,500-4,500 pages)

6. Getting Started
• Product Development Strategy should incorporate:
– Manufacturing
– Preclinical Pharmacology and Toxicology
– Clinical Plans (from FIH to Phase II)
– Regulatory
– Remember that all aspects are inter-related
• Budget/Resource Management

7. Approaching the Nonclinical Safety Testing
Program
• Toxicology studies should not be considered a box-
checking exercise to simply satisfy Regulators
– Contributes to the understanding of the product
– Provides the supporting data to enable FIH studies (target
organs, predict toxicology, reversibility, exposure levels,
starting doses, etc.)

8. Perspective
• Preclinical development is an expensive investment
for a small/emerging company that requires:
– A good plan/strategy that considers regulatory
expectations and the Company’s objectives
(scientific/medical and business)
– Efficient and expedient implementation by experienced
individuals
– Interpretation and positioning of results by experts
• If not designed, conducted, and/or interpreted
correctly, preclinical studies can add considerable
time and expense to a program

9. General Toxicology Program Considerations
• Regulatory expectation (21 CFR Part 312): Nonclinical
safety studies should be conducted to determine the
safety of proposed clinical trials
• Studies should be Conducted in accordance with Good
Laboratory Practice (GLP) regulations/principles
– An international quality standard
– It’s about the documentation!
– Covers personnel, facilities, equipment, operations, test article,
data entry, reports, etc.
– Does not cover interpretation or evaluation of data
– Contributes to the timing and expense of studies
– Sponsor has obligations
› Ensuring the integrity of the data - monitoring the study

10. General Toxicology Program Considerations
(cont’d)
• The preclinical development plan will depend on a
number of factors, including:
– Product type and similarity to existing agents with known
safety profiles
– Proposed indication in humans (i.e., cancer vs rheumatoid
arthritis)
– Proposed duration of administration (i.e., short term vs
chronic; dosing regimen)
– Target population (i.e., adults, infants, pregnant women,
elderly, etc.)
– Proposed route(s) of administration
– Use pattern considerations (i.e., concomitant medications,
adjuvant therapy)

11. General Toxicology Program Considerations
(cont’d)
• Studies should be designed specifically for the drug
under development
– Relevant animal species (i.e., pharmacologically active)
› Particularly for biologics / therapeutic proteins
– Knowledge of expected toxicities
› Dose range finding data and pilot studies
› Drug class effects (published literature, Freedom of
Information, E.U. EPARs, scientific meetings, etc.)
› First principles
› Interaction with regulatory authorities

12. Objectives of Early Toxicology Studies
• Identify the target organs / systems of the drug
– Monitoring in clinical trials
– Gender differences
– Expected? (based on pharmacology)
• Characterize the dose-response curve
– No-Observed-Adverse-Effect Level (NOAEL)
› Important for Maximum Recommended Starting Dose (NCE)
– Maximum tolerated dose (MTD)
– Therapeutic Index

13. Objectives of Early Toxicology Studies (cont’d)
• Characterize the toxicity
– Reversible?
– Dose-dependent?
• Assess the systemic exposure
– Calculate pharmaco-/toxicokinetics (Tmax, Cl, Vd, t½)
– Margins of safety relative to human exposures based on
AUC and Cmax
– Aid in dose selection for further animal toxicology, FIH
studies

14. Developing a Biologic is Different From a Drug
Differences between small molecules and biologics
– a generalization
Small Molecule Drug Biologic
Low molecular weight High molecular weight
Familiar antecedents Potentially unique
Known impurities Unfamiliar impurities
Often orally dosed Often parenteral, IV dosing
Maximal tolerated dose Optimal biologic dose
Meaningful chronic tox Uncertain chronic tox
Species-independent Species-specific
Biotransformed Degraded
Not immunogenic Immunogenicity issues

15. Typical IND-Enabling Preclinical Safety Studies
Species Duration of Studies Cost ($)*
(Bio)analytical Assay development 1,000/day
Validation (per species) 15,000-20,000
Running samples 70-100/sample
Dose formulation Analyses $5K/time study
Rat Single dose 29,000-75,000
7 day DRF 50,000-125,000
14 days 165,000-200,000
28 days 120,000-275,000
Dog Maximum tolerated dose (MTD) 30,000-65,000
7 day DRF 75,000-145,000
14 days 140,000-300,000
28 days 200,000-450,000
* Pricing will vary depending on the actual study design, route of administration,
numbers of groups, numbers of animals, bioanalytical determinations, special
tests required, etc.

16. Typical IND-Enabling Preclinical Safety Studies
(cont’d)
Species Type of Studies Cost ($)*
Monkey MTD 75,000-125,000
7 day DRF 100,000-240,000
14 days 265,000-410,000
28 days 280,000-550,000
Genetox Bacterial mutagenicity 6,000-8,000
Chromosome aberration 20,000-27,000
Rodent micronucleus 25,000-35,000
Safety Pharm hERG inhibition (patch clamp) ~16,000
CNS rodent 25,000-35,000
Cardiovascular (telemetry) 75,000-120,000
Respiratory 25,000-35,000
* Pricing will vary depending on the actual study design, route of administration,
numbers of groups, numbers of animals, bioanalytical determinations, special tests
required, etc.

17. Typical IND-Enabling Preclinical Safety Study
Generalized design of a repeated-dose rodent toxicity study
Treatment Dose Main (Terminal) Recovery Toxicokinetics*
Group (mg/kg/day) Male Female Male Female Male Female
Vehicle 0 10 10 5 5 0 0
Low Dose A 10 10 0 0 9 9
Mid Dose B 10 10 0 0 9 9
High Dose C 10 10 5 5 9 9
• Number of animals depends on blood volumes required, number
of timepoints, etc.
• GLP-compliance needed for a pivotal (clinical trial-supporting)
study
• Toxicokinetic evaluations
• Standard AND drug- or disease-specific endpoints
• Histopathology

18. Example 1: (NCE, Cancer Indication)
• Program
– Abbreviated, in 2 species
– Focus on repeat-dose studies (cycles of treatment)
• Issues
– Multiple salt forms (available at different times)
– Dose selection in studies (cytotoxic NCE for cancer, so
based on STD10, not NOAEL)
– 2 very different routes of administration (mechanisms of
action dependent on route)

19. Example 2 (Biologic, Non-cancer Indication)
• Program
– Full program, 2 species
– Molecular target (human)
• Issues
– Target down-regulated in healthy/normal animals/humans
(toxicology program in healthy animals? Or disease
models?)
– Test material quality (research/pre-GMP/GMP)
– Need to consider efficacy data when setting doses in
humans (MABEL approach), not just toxicology data
– Need to evaluate immunogenicity (toxicology program;
method development)

20. Strategies for Success
• Desired: a successful preclinical development program
that results in a high quality regulatory submission that
anticipates the questions of the Regulator
• Understand that the animal toxicology data impact the
clinical trial design, but the trial design can also
influence the toxicology program
• Co-ordinate Toxicology and Manufacturing
– Ensure that high-quality (ideally, GMP) product is available
(quantity, timing, quality/identity/purity/stability is documented)
• Have a regulatory strategy
– Pre-IND/Pre-CTA consultation?
– IND submission preparation co-ordinated with timing of
toxicology program?

21. Strategies for Success (cont’d)
• Become “IND-Ready”, even at the preclinical stage
of development:
– Pharmacology studies: proper reports (report numbers,
etc., and consider electronic filing)
– Regulatory toxicology considerations (GLP compliance,
etc.)
– Anticipate types of data that will be needed
– Manufacturing considerations (timing, logistics, stability
data, characterization and release of API/product, with an
eye to GMP product for trials)

22. Strategies for Success (cont’d)
• Present a high-quality submission: avoid
Regulatory “Red Flags” and address reviewer’s
expectations (content, format, quality of data to
support the proposed trial, etc.)

23. Help Needed!
• Regulatory Agency guidelines (Canada, US, EU)
• Regulatory Agency consultations (Pre-IND or Pre-
CTA meetings)
• Published commentaries, opinions, and guidance
• Meetings, Conferences, Colleagues
• Consultants

24. FDA Guidance Related to Preclinical Safety
Evaluation
• FDA Regulatory Pharmacology and Toxicology
homepage:
• www.fda.gov/cder/pharmtox/default.htm
– Links to all FDA guidance documents (draft and final)
› ICH
› FDA
– Internal policies and procedures
– Contact names of Pharm/Tox staff within CDER
• Other sources (examples)
– Oncology Drugs - DeGeorge, et al., 1998. Regulatory
consideration for preclinical development of anticancer drugs.
Cancer Chemother Pharmacol 41:173-185.
– Inhalation Drugs - DeGeorge, et al., 1997. Considerations for
toxicology studies of respiratory drug products. Regul Toxicol
Pharmacol 25:189-193.

25. THANK YOU !
Valentia Lee-Brotherton, PhD
Ashuren Health Sciences
www.ashuren.com