Paving the way for Non-Clinical Bioanalytical Partnerships Louise Angell

Transcription

1 Paving the way for Non-Clinical Bioanalytical Partnerships Louise Angell

2 Content Overview of non-clinical immunogenicity testing for biologics Regulatory guidance Bioanalytical considerations Risk based approach CRO Pharma

3 Overview of Immunogenicity Testing All biologics (recombinant therapeutic proteins) will induce an immune response Immunogenicity can be either wanted (vaccine) or unwanted Unwanted ADA Binding (Ab binds to the drug but has no apparent impact of PK and PD) Sustaining (Ab binds to the drug and prolongs its half-life, increasing drug exposure) response has potential safety Clearing (Ab binds to the drug and accelerates clearance from implications circulation, decreasing drug exposure) Neutralizing (Ab binds to the drug and prevents pharmacological activity)

4 Overview of Immunogenicity Testing Test during non-clinical and clinical programs Immunogenic response in non-clinical study is not predictive of an immune response in humans Novel Therapeutic Protein May assume humanised protein to have less immunogenicity risk than prokaryotic engineered protein Humanised protein will have greater homology to native sequence than non-humanised, however immunogenicity frequency is still variable Understand BioCMC and in vitro data Assess in repeat dose non-clinical studies, multiple species to enable interpretation of non-clinical data

5 Regulatory Guidelines - Summary Regulatory agencies in the US and EU are consistent in their recommendation that immunogenicity be evaluated from a patient safety perspective due to non-predictability of data from non-clinical studies Immunogenicity testing is required for novel compounds Regulatory guidance for bioanalytical method validation is well defined for PK assays, with multiple white papers published for ADA assays, although ADA interpretation is evolving For Biosimilar products, there is no global guidance for immunogenicity testing, and EMA infer in vivo testing is not required due to lack of predictability in vivo studies may be considered reasonable markers for potential immunogenicity assessment due to differences in product specification (quality and manufacturing)

6 Regulatory Limitations Potential for differing interpretation of the guidelines by companies Biosimilar (ADA Assays) Criteria for comparison of data? It is not clear if one assay or two assay approach is optimal Reference material? One assay - additional assessments (cut point and drug tolerance?) Pre-existing antibodies There is no regulatory guidance on how to deal with this

7 Bioanalytical Considerations

8 Overview of ADA assay anti-drug antibody (ADA) labelled drug biotin-labeled drug Streptavidin coated plate

9 Response Overview of ADA assay Qualitative screening assay, confirmation, titre quantification Mean Cut Point -ve samples +ve samples 5% False Positives Assay signal

10 Insulin BioSimilar Case Study EMA Guideline on non-clinical and clinical development of similar biological medicinal products containing recombinant insulin and insulin analogues (draft April 2014) Risk based approach for toxicology studies Presence of pre-existing antibodies against insulin is well documented Anti-Drug Antibody Capture reagent Anti-insulin antibody biotin-labeled drug Streptavidin coated plate

11 Insulin Biosimilar Case Study Cut point set using naïve individuals however this included positive samples Assay sensitivity was high (close to 1000ng/mL) Some samples confirmed positive with both biosimilar and insulin Potential to falsely report naïve background samples as confirmed positive At risk of missing low concentration, true positives with high affinity binding Many hours of Client communication to discuss concerns and risks Scientists came together to discuss potential issues Rat sample analysis study: ~5% samples screened positive and no impact upon TK profile Learnings Buffer cut point (sample with background greater than buffer would be positive) Pre-incubate cut point sample with insulin to remove pre-existing antiinsulin Ab whilst capturing biological matrix variation Skip confirmation step Titre pre and post dose for comparison

12 Non-Clinical Study Design Some Clients have well defined BioCMC package and good prediction of immunogenicity risk Other Clients have limited experience with developing biologics Dosing Human protein into mouse led to a high level of immunogenicity (60%) Non-clinical program designed in 1 species Immunogenicity response could not be correlated to the decrease in TK profile because ADA samples were collected from different animals to TK samples PD data subsequently generated to justify administration of human protein in mouse model was responsible for apparent immunogenicity Enhanced relationships help to guide Subsequently, ELISA method transferred and validation using Gyros to generate ADA data from TK samples. This projects confirmed TK reduction correlated to ADA response These data were used to support absence of 2 nd rodent model Cyno study later demonstrated reduced/no immunogenicity

13 Non-Clinical Study Design Learnings for non-clinical sample collection and testing Importance of pre-dose samples Align ADA with TK sampling Collect enough sample to allow tier testing (screen, confirmation, titer, nab) Design to account for sample volume limitations (rodents) Choice of bioanalytical assay Drug interference (extend wash-out period so drug is BLQ)

14 Bioanalytical Considerations ADA In-Lab Analytical Time Transfer Development 1-2 weeks In 2014, ADA transfer studies lasted an average 6 week in-life period 2-3 weeks Validation Not just analytical time, time is required 3-4 weeks for data review, Client discussions, Client internal debate Transfer Innovator + Biosimilar What are Client (submission) timelines Development Innovator + Biosimilar Validation Innovator + Biosimilar 1-2 weeks 2-3 weeks Additional time for positive control generation 3-5 weeks

15 Improved Communication Who is involved in a non-clinical drug development program? DMPK Client Study Monitor Bioanalysis BioCMC Non-Clinical Are Program communications well defined? QA Manufacturing Safety Assessment Discovery group

16 Risk Based Approach for ADA Testing

17 Interpretation of Immunogenicity Immunogenicity must be considered with other endpoints ADA onset CRO challenge of immune to generate response data & change in PK PK.pharmacokinetics (drug exposure) PD.pharmacodynamics Pharma challenge to consider (drug activity) all data and make AE.adverse immunogenicity events/observations assessment (drug toxicity).all Pharma are important could increase when interpreting efficiency study of CRO results! by ensuring data are shared ADA + PK + PD + AE = Immunogenicity Assessment

18 Risk Based Approach for ADA Testing Regulatory agencies consistently recommend that immunogenicity be evaluated from a patient safety perspective As such, companies should utilize a risk-based approach in evaluating potential immunogenicity of their drug products Immunogenicity is difficult to predict Potential immunogenicity of the protein Biological function of the protein Endogenous counterparts Route of administration Dose and frequency of administration Health status of subjects

19 Bioanalytical Cost Limitations Is ADA testing required? Is a fully validated method required? Trastuzumab (Herceptin Biosimilar) Supported several full validations (~3 weeks) Assay sensitivity can be affected by drug tolerance Use risk based approach to understand BioCMC and in Acid vitro dissociation data can affect sensitivity Plan in vivo studies and consider commitment to Is it necessary ADA assay to change non-clinical design? Cannot confirm ADA response in presence of several mg drug Typical ADA sampling 0, 1, 4, 13 weeks IgG t ½ = 6 weeks after 8 day response time Costly to include wash out periods (TK trough samples)

20 Summary Regulatory agencies in the US and EU are consistent in their recommendation that immunogenicity be evaluated from a patient safety perspective, however this is open to interpretation CROs can be well placed to advise Clients of potential bioanalytical limitations Pre-existing antibodies Non-clinical study design Consideration to bioanalytical timelines Increased communications between Pharma and CRO can benefit Use risk based approach to consider extent of bioanalytical testing required

21 Thank You

22 Additional info for reference as required

23 ICH Regulatory Guidelines? Many biotechnology-derived pharmaceuticals intended for human are immunogenic in animals. Therefore, measurement of antibodies associated with administration of ICH S6 Preclinical Safety Evaluation of Biotechnology-Derived Pharmaceuticals (R1) these types of products should be performed when conducting repeated dose Q5E toxicity Comparability studies in order of Biotechnological/Biological to aid in the interpretation of these Products studies. Subject Antibody to Changes responses in Their should Manufacturing be characterised Process (e.g. titer, number of responding animals, Q2B Guideline Validation of Analytical Procedures Methodology neutralising or non-neutralising), and their appearance should be correlated with any pharmacological and/or toxicological changes. US Animal immunogenicity assessments generally do not predict potential immunogenic responses to protein products in humans. However, when differences in manufacturing (e.g., impurities or excipients) between the proposed product and Immunogenicity Assessment for Therapeutic Protein Products, August 2014 the reference product may result in differences in immunogenicity, measurement of anti-protein antibody responses in animals may provide useful information relevant Scientific Considerations in Demonstrating Biosimilarity to a Reference Product, to patient February safety (draft) Bioanalytical Method Validation, September

24 Regulatory Guidelines? EU Immunogenicity Assessment of Biotechnology-Derived Therapeutic Proteins, 2007 (revision due) Guideline on Similar biological medicinal products containing monoclonal antibodies: non-clinical and clinical issues, 2010 Guideline on Similar biological medicinal products containing biotechnology-derived proteins as active substance: non-clinical and clinical issues, 2005 Guideline on Bioanalytical Method Validation (2011)..the predictivity of non-clinical studies for evaluation of immunogenicity is considered low. Nonclinical studies aiming at predicting immunogenicity in humans are normally not required. However, ongoing consideration should be given to the use of emerging technologies which might be used as tools. Measurement of antibodies in non-clinical studies are however requested as part of repeated dose toxicity studies, in order to aid in the interpretation of these studies.. the comparison of the antibody response to the reference product in an animal model may be part of the comparability exercise both for similar biological medicinal products. and for changes in manufacturing Due to the different production processes used by the biosimilar and reference product manufacturers, qualitative differences of process related impurities will occur... Qualitative or quantitative difference(s) of product-related variants may affect biological functions of the mab and are expected to be evaluated by appropriate in vitro assays. These quality differences may have an effect on immunogenic potential and potential to cause hypersensitivity. It is acknowledged that these effects are difficult to predict from animal studies and should be further assessed in clinical studies. Immunogenicity assessment in animals is generally not predictive for immunogenicity in humans, but may be needed for interpretation of in vivo studies in animals. Blood samples should be taken and stored for future evaluations if then needed. Non-clinical toxicity as determined in at least one repeat dose toxicity study, including toxicokinetic Copyright 2014 Covance. measurements. All Rights Reserved 24

25 Immunogenic Classes of Therapeutic Proteins Strong Class Description Human Protein Homology Immunogenicity Frequency Examples A Prokaryotic Low High Staphylokinase B Mammalian Low High OKT-3 Weak C D E F Novel Construct Medium Variable High: Denileukin Low: human growth hormone Chimeric Human High Variable H: chmul6 L: rituximab Humanized High Variable L: Campath-1 H: Human anti- CD3 Human Identical Variable H: GM-CSF L: Human insulin Bugleski Copyright 2014 and Covance. Treacy, All Rights Reserved Cur Opinion Mol Ther 6: 10-16, 2004

26 Animal Data Not Predictive Protein Therapeutic Streptokinase and Staphlyokinase Preclinical Immunogenicity? High Clinical Immunogenicity? High Keyhole Limpet Hemocyanin High in rodents High Human interferon α- 2a High in rodents Low Human Growth Hormone High in rodents Low Human Interferon-λ High in Cynomolgus monkeys Low Human Interleukin-3 High in Rhesus monkeys Low Bugleski and Treacy, Cur Opinion Mol Ther 6: 10-16, 2004

27 Estimated Risk and Study Design Risk Level Drug Characteristics Examples Consequences Low Not structurally identical to endogenous protein Non agonistic Enzymes Antibody drugs Infusion site reactions Loss of efficacy Mild allergic reactions Medium Partially or completely identical to endogenous protein Endogenous counterpart is either missing or redundant or Not structurally identical to endogenous protein/agonistic Replacement therapy, such as Factor VIII Antibody drugs Same as Low Risk Overstimulation of endogenous mechanism Immune complex formation High Partially or completely identical to endogenous protein Endogenous counterpart is not redundant Erythropoeitin GM-CSF Same as Medium Risk Neutralization of endogenous counterpart

28 Estimated Risk and Study Design Bioanalytical Testing Strategy Risk Level Sampling Frequency Assessment ADAs Low Medium High More frequently earlier in development, less frequently in Phase III (baseline, end of study and possibly follow-up) More frequently earlier in development, less frequently in Phase III (baseline, end of study and possibly follow-up) More frequently throughout all phases of clinical trails. Consider real-time testing of ADA and Nab Screen / Confirm Titer assessment Further characterization may be helpful Nab, Mapping ADA Screen / Confirm Titer assessment Further characterization may be helpful Nab, Mapping ADA Screen / Confirm / Titer / NAb Further characterization of ADA by mapping, isotyping etc. Sequential patient dosing rather than cohort model for first-in-human studies Rosenberg and Worobec; Biopharm. Int. 17, 17, 18 (2004)