1. Anne M. Pilaro, Ph.D. Center for Drug Evaluation and Research FDA
Nonclinical Safety Testing for Biological Therapeutics for Cancer Treatment
Anne M. Pilaro, Ph.D.
Center for Drug Evaluation and Research
FDA

2. Disclaimer
The opinions expressed by Dr. Pilaro in this presentation are the presenter’s, and do not reflect official policy of the FDA
Information presented was obtained from publicly available sources, including published literature, labeling, and/or SBA documents
No official support or endorsement by FDA is intended or should be inferred

3. Objectives for Today’s Presentation
Discuss review responsibilities for biological therapies for cancer
Review approaches to obtain IND-enabling, nonclinical safety data for protein therapeutic agents for cancer
Introduce current guidance for toxicology testing of biological therapeutic products

4. What is a Biological Product?
Substances derived either from living organisms – including humans, animals, plants, and microorganisms – or produced by biotechnology methods
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5. Who is Responsible for Regulating Biologic Cancer Therapies?
CBER, OCTGT
adoptive cell transfers (i.e., stem cells, embryonic stem cells [ESC], leukocytes with or without gene modification)
gene therapies
tumor vaccines (e.g., pDNA-based, viral vector-based, or autologous/allogeneic tumor with or without gene modification)
siRNAs delivered using gene transfer technology
combination therapies

6. Who is Responsible for Regulating Biologic Cancer Therapies?
CDER/OODP/DDOP
hormones and metabolic factors
synthetic peptides
oligonucleotides and siRNA
CDER/OODP/DBOP
monoclonal antibodies
recombinant proteins
e.g., cytokines, growth factors, fusion proteins
extracted proteins

7. Who is Responsible for Regulating Biologic Cancer Therapies?
For the purposes of this presentation, focus today will be on products regulated in CDER/OODP/DBOP
monoclonal antibodies for cancer treatment
cytokines and/or growth factors
e.g., interleukin-2 for cancer treatment
recombinant hematopoietic factors for treatment and/or prophylaxis of chemotherapy side effects
fusion proteins for cancer treatment
combination antibody~ or protein~small molecule drug conjugates

8. What Do I Need to Start an IND for a New Anti-Cancer Agent?
Product characterization
manufacturing and quality control issues
Nonclinical testing in animals
biologic or pharmacologic activity
pharmacokinetic profile
toxicology testing in animals
Phase 1 clinical trial

9. What are Pharmacology Studies for Anti-Cancer Biologics?
Evaluation of ability of a new agent to induce the desired therapeutic effect
in vitro studies of product binding, tumor cell killing, and other effects
in vivo studies of anti-tumor activity
e.g., human tumor xenograft models
Demonstration of pharmacologic and/or biologic activity is the first step in the development of ANY new drug or biologic

10. Why are Pharmacology Studies Important?
Establish basis for conducting trial
interaction with target receptor, tissue, or organ
degree of pharmacodynamic (P/D) effect
correlation of P/D effect with exposure
Optimize dosing, regimen for the clinic
Optimize route of administration
Selection of species for further testing

11. Where do Pharmacology Data Come From?
Most biologic activity/pharmacology data come from studies conducted in the sponsor’s own laboratory
need to include appropriate control groups
should include sufficient “n” in treatment groups to attain statistical relevance
should be described in the IND in adequate detail that FDA can judge appropriateness of model, findings to proposed human study
Published literature may also serve as data source
data in article must be of sufficient detail, as above
For previously tested agents, access to data may be obtained by permission to cross-reference existing IND or DMF

12. How are Pharmacology Data Used to Support a New IND?
Data from nonclinical bioactivity and/or pharmacology studies are used to establish the rationale for conducting the first clinical trials in humans
for anti-cancer biologic agents, most early trials are done in the index (patient) population, NOT healthy volunteer subjects

13. What are Pharmacokinetic Studies for Anti-Cancer Biologics?
Measurement of in vivo disposition of a new biologic agent
studies of product concentration, exposure, clearance, and half-life
determined from blood, plasma, or serum levels of agent at various times after dosing
differences following different routes of dosing

14. Why are Pharmacokinetic Studies Important?
Demonstration of the pharmacokinetics of a new biologic allows estimation of:
exposure to agent after any given dose
correlation with pharmacologic/therapeutic effect
duration of exposure (half-life)
dosing interval for the clinical study
time to reversal of any biologic or toxic effects
development of anti-product antibodies
both total and neutralizing activity

15. Where do Pharmacokinetic Data Come From?
Serum levels of anti-cancer biologic measured during in vivo pharmacology studies
Stand-alone pharmacokinetic studies in responsive species
Adjunct to single- and repeat-dose toxicology studies in animals
toxicokinetics

16. What are Toxicology Studies for Anti-Cancer Biologics?
Dedicated in vivo studies that provide data describing the safety profile of the agent, at doses greater than those that produce the therapeutic effect
single- and repeat-dose paradigms
sub-chronic and chronic duration
include measurements of exposure, antibody development, and reversibility

17. Why is Toxicology Testing for Any New Biologic Important?
Provides information regarding the safety of single and/or repeated exposure to the biologic anti-cancer agent
toxicities related to the dose of product given
toxicities related to the duration of product administration
identification of target organs
reversibility of toxicities
after clearance of the biologic product
after development of neutralizing antibodies

18. What Does a Nonclinical Toxicology Study Tell Me?
Single dose (acute) studies
designed to maximize exposure to agent (mimic overdose)
high doses, frequently i/v administration
limited endpoints – 14 days
clinical observations, body weights
gross necropsy with no histology
clinical pathology not usually done

19. What Does a Nonclinical Toxicology Study Tell Me?
Repeat-dose administration
Designed to support the intended clinical usage of the agent
doses “bracket” those planned for clinic
dosing for at least the same number of planned administrations
duration may vary from 14 days to 6+ months
should include recovery group(s)

20. Where do Toxicology Data Come From?
Data in support of a clinical trial may come from:
GLP-compliant toxicology studies conducted by a contract laboratory
well-controlled studies conducted in house (“GLP-like”)
published data in peer-reviewed journals
cross-reference to previously submitted IND or DMF

21. What Do I Need to Measure in Toxicology Testing?
Monitoring in-life
clinical observations, behavior
signs/symptoms of toxicity
body weights, weight changes
feed and water consumption
clinical pathology
serum enzyme, electrolyte, glucose and lipid profiles as evidence of organ toxicity
hematology
urinalysis
these can all also be monitored in man!

22. What Do I Need to Measure in Toxicology Testing?
Pharmacokinetics/toxicokinetics
correlation of serum levels of biologic with toxicity, pharmacodynamic activity
identification of elimination half-life and antibody development (reversibility)
Pathology
gross and microscopic evaluation of tissue damage
most sensitive indicator, but final!
confirmation of target organ toxicity detected biochemically

23. What Else Do I Need to Include in My Toxicology Studies?
Administration/dose selection
route & dosing regimen should mimic proposed clinical use
alternative routes/regimens acceptable in some cases
attainment of toxic dose, no observable adverse effect level (NOAEL) desirable
multiples of human dose needed to determine adequate safety margins
can vary with product class and/or clinical indication

24. Are There Any Specific Safety Concerns for my Biologic Agent?
Monoclonal antibodies (naked or conjugated)
cross-reactivity with normal tissue
exaggerated pharmacodynamic effects
chronicity of exposure and toxicity
immunogenicity/antibody production
“bystander” toxicity of radiolabel or small molecule drug in conjugates

25. Are There Any Specific Safety Concerns for my Biologic Agent?
Cytokines and/or growth factors
demonstration of species-specificity
evaluation of interaction with host endogenous cascade
tumor-promoting potential
immunogenicity/antibody production
effects on neutralization of endogenous counterpart to test agent

26. How Are Toxicology Studies Used to Support a New IND?
Nonclinical toxicology data are used to make recommendations regarding the conduct of the clinical trial, based on the safety data obtained
initial safe starting dose, dose-escalation scheme
target organ(s), reversibility of toxicity
appropriate parameters for clinical monitoring
identify “at risk” patient populations (inclusion/exclusion criteria)

27. What Else Do I Need to Know for Safety Testing of Biologics?
Data from preclinical pharm/tox studies must meet the regulatory requirements set forth in 21 CFR
data must be of unquestionable integrity, quality to support safety of agent for entry into clinical trials
animal studies may be done in house as long as meet the requirements of 21 CFR (8)
full compliance with GLP (21 CFR 58) expected for any pivotal toxicity studies

28. What Else Do I Need to Know for Safety Testing of Biologics?
Product development allowed during preclinical testing phase
use of non-GMP protein products allowed
need to demonstrate comparability with clinical grade material(s) as development progresses
Protein therapeutics are not expected to differ from small molecule cancer drugs in the requirements for demonstration of safety, just in the approach used to obtain the data

29. Where Can I Find Guidance for Safety Testing of Biologics?
The ICH S6 Document
Title: Preclinical Testing of Biotechnology-Derived Pharmaceuticals
Scope: Covers protein and other biological therapeutics produced by biotechnology methods
Does not address: cellular, tissue or gene therapies, blood or plasma products, traditional vaccines 34

30. ICH-S6 – What it Doesn’t Do
Does NOT provide a “one size fits all” or “cookbook” approach to toxicology study design
provides a framework for design of animal studies to address safety of biotechnology derived products, based on characteristics of the product and intended clinical use 26 42 26

31. ICH-S6 – What it Does Do
Recognizes that conventional approaches to toxicity studies used for small molecule drugs are often NOT appropriate for protein therapeutics
unique issues of species restriction, dose and formulation limitations, antibody formation limit utility of standard toxicity paradigms
offers alternative approaches to obtain adequate safety data to support human use 26 42 26

32. ICH-S6 – What it Does Do
Provides guidance to sponsors regarding the design of toxicology studies, including:
usage of pharmacologically relevant vs. nonrelevant animal species
usage of animal models of disease
immunogenicity testing and its implications
genotoxicity, chronic toxicity, carcinogenicity, and reproductive and developmental toxicity testing
GLP compliance 26 42 26

33. Additional Guidance from FDA
Coming soon…
Guidance for Industry and Reviewers: Nonclinical Safety Evaluation of Biotechnology-Derived Pharmaceuticals
FDA general guidance on necessary studies to support development of protein therapies
expands upon the framework of ICH S6
adds FDA experience with unique issues
includes Q&A section to address common questions encountered by sponsors in field 31 47 31

34. Additional Guidance from FDA
Coming soon…
Guidance for Industry, Investigators and Reviewers: Nonclinical Studies for Anticancer Drugs and Biologics
OODP-specific guidance on necessary studies to support development of anti-cancer therapies
identifies differences in nonclinical study approaches for small molecule versus protein cancer therapeutics
addresses issues of timing of studies relative to duration of clinical trial, indication
incorporates principles of both ICH S6 and FDA Guidance for protein therapeutics 31 47 31

35. Summary
What do I need to start an IND for a new anti-cancer biologic agent?
The “Three R’s” of nonclinical testing:
pharmacology = RATIONALE
toxicology = RECOMMENDATIONS
demonstration of safety = REGULATORY EXPECTATIONS

36. Take-Home Messages
Animal testing determines safety…but caveats exist for biological products
Toxicology programs for anti-cancer biological therapeutics require novel approaches to obtain data
no “one size fits all” paradigm for biologics
traditional animal toxicology models may not be appropriate or feasible
studies may have to be “individualized” to address specific safety concerns

37. Take-Home Messages
Toxicology studies for anti-cancer biological therapeutics should:
follow guidance set forth by ICH S6, and additional guidances as available
be rational, scientifically designed, and problem solving
employ careful design, judicious use of animals

38. Take-Home Messages
But... animal studies really only give you a “best guess” of what to expect
no animal model, including humans is 100% predictive of response in man
sometimes animal studies can give a false sense of security
however, these models may be useful in evaluation of mechanism, other toxicities

39. Some Further Resources
ICH Guidances
ICH S6: Safety Studies for Biotechnological Products
ICH M3: Timing of Pre-clinical Studies in Relation to Clinical Trials
ICH S5a: Detection of Toxicity to Reproduction for Medicinal Products
ICH S2b: Standard Battery of Genotoxicity Testing
also available at:

40. Some Further Resources
Points to Consider
Points to Consider in the Manufacture and Testing of Monoclonal Antibody Products for Human Use - 2/28/97
Points to Consider in the Manufacture and Testing of Therapeutic Products for Human Use Derived from Transgenic Animals – 1995

41. Further Questions? Please Contact the DBOP Toxicologists
Anne M. Pilaro, Ph.D.
Supervisory Toxicologist
DBOP Staff
Andrew McDougal, Ph.D.
Brenda Seidman, Ph.D.
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