Bioanalytical Support to In Vitro Studies

Transcription

1 Bioanalytical Support to In Vitro Studies Presenter: Tim Sangster (on behalf of EBF (mini-workshop)) Open Symposium 2017 November 17 th 2017 Barcelona

2 Agenda Ø Scope Ø Aims Ø Define GLP Ø In Vitro Submission Requirements Ø Proposed Strategy Ø Case Study 2

3 Scope Ø mws held in EBF Strategy meeting in Limelette on the 15 th March 2017 Ø Discussion on supporting in vitro assays and the appropriate quality standards and approach Ø Clear split many companies doing GLP for assay validation and live study Ø Agreement that no clear guidance is available Ø Agreement that we do more than required 3

4 Aims Ø Outline the current requirements for the BioA work on in vitro studies Ø Propose a workable approach for supporting in vitro studies 4

5 Defining GLP OECD Q&A Ø 1)What are the OECD Principles of Good Laboratory Practice (GLP)? Ø a) The Principles of Good Laboratory Practice (GLP) are a managerial quality control system covering the organisational process and the conditions under which non-clinical health and environmental studies are planned, performed, monitored, recorded and reported. The OECD Principles of GLP are followed by test facilities carrying out studies to be submitted to national authorities for the purposes of assessing the health and environmental safety of chemicals and chemical products which may also be of natural or biological origin and, in some circumstances, may be living organisms. Depending on the jurisdiction, the Principles of GLP can also be applied to nonclinical safety testing of other regulated products, such as medical devices. 5

6 Defining GLP OECD Q&A Ø 1)What are the OECD Principles of Good Laboratory Practice (GLP)? Ø b) The Principles of GLP define the responsibilities of test facility management, study personnel and quality assurance personnel that are operating within a GLP system, and minimum standards concerning the suitability of facilities and equipment to perform studies, the need for standard operating procedures, documentation of raw data, study reports, the archiving of records, etc 6

7 In Vitro Submission Requirements ICH S2(R1) Ø Genotoxicity tests can be defined as in vitro and in vivo tests Ø Assessment of mutagenicity in a bacterial reverse gene mutation test. This test has been shown to detect relevant genetic changes and the majority of genotoxic rodent and human carcinogens. Ø Genotoxicity should also be evaluated in mammalian cells in vitro/or in vivo Ø No real BioA work on these studies. 7

8 In Vitro Submission Requirements, cntd Ø herg has become a key part of the safety pharmacology package requirements as discussed in the ICH S7B Ø Analysis of the compound concentration in the test system is an important criteria. 8

9 In Vitro Submission Requirements, cntd ICH S7A Ø It is important to ensure the quality and reliability of non-clinical safety studies. This is normally accomplished through the conduct of the studies in compliance with GLP. Due to the unique design of, and practical considerations for, some safety pharmacology studies, it may not be feasible to conduct these in compliance with GLP. It has to be emphasized that data quality and integrity in safety pharmacology studies should be ensured even in the absence of formal adherence to the principles of GLP. 9

10 In Vitro Submission Requirements, cntd ICH M3(R2) Ø In vitro metabolic and plasma protein binding data for animals and humans and systemic exposure data (ICH S3A, Ref. 7) in the species used for repeated-dose toxicity studies generally should be evaluated before initiating human clinical trials. Ø Further information on pharmacokinetics (PK) (e.g., absorption, distribution, metabolism and excretion), in test species and in vitro biochemical information relevant to potential drug interactions should be available before exposing large numbers of human subjects or treating for long duration (generally before Phase III). These data can be used to compare human and animal metabolites and for determining if any additional testing is warranted. 10

11 In Vitro Submission Requirements, cntd ICH M3(R2) Ø In addition, primary PD studies (in vivo and/or in vitro) are intended to investigate the mode of action and/or effects of a substance in relation to its desired therapeutic target. Such studies are generally conducted during the discovery phase of pharmaceutical development and as such, are not generally conducted in accordance with Good Laboratory Practices (GLP). These studies can contribute to dose selection for both nonclinical and clinical studies. 11

12 In Vitro Submission Requirements, cntd ICH M3(R2) Ø Studies providing acute toxicity information can be limited to the clinical route only and such data can be obtained from non-glp studies if clinical administration is supported by appropriate GLP repeated-dose toxicity studies. Ø In some specific situations (e.g., microdose trials; see Section 7) acute toxicity or singledose studies can be the primary support for studies in humans. In these situations, the high dose selection can be different from that described in Section 1.5 but should be appropriate for supporting the intended clinical dose and route. These studies should be performed in compliance with GLP. 12

13 In Vitro Submission Requirements, cntd ICH M3(R2) Ø Table 3 Recommended Non-Clinical Studies to Support Exploratory Clinical Trials Ø Footnote a. General toxicity studies should be conducted according to GLP regulations. 13

14 In Vitro Submission Requirements, cntd EMA Guidance Ø This guideline provides recommendations for the validation of bioanalytical methods applied to measure drug concentrations in biological matrices obtained in animal toxicokinetic studies and all phases of clinical trials. 14

15 In Vitro Submission Requirements, cntd Ø GLP is clearly identified in the regulatory documents for specific study types Ø Bioanalysis is not mentioned specifically Ø EMA guidance for Bioanalysis does not cover the in vitro assays 15

16 Proposed Strategy Ø Understanding the experimental system and the whole experiment to be conducted in the context of BioA requirements Ø Assay Validation A&P Stability etc Ø Process Validation have we tested the conditions throughout the process? Ø Design the appropriate controls 16

17 ????? A valid Assay???? How do we define a valid assay today, in view of the increased scope and variety of bioanalysis? Can this only be achieved using regulatory standards as per Guidance Is there a valid alternative, i.e. less resource demanding, allowing better scientific focus? 27/11/2017 htt-sp:// 17

18 Proposed Strategy Tiered approach 18

19 Fit for purpose = general term, often used in BM assays Tiered approach Different levels of quality in early discovery assays. Refer to EBF or GBC paper and relates to research/screening assays Scientific validation Refer to 2015 EBF Recommendation paper and relates to 5 assay types where industry tends to use regulatory validation 1. Urine 2. tissue homogenate 3. pre-mad metabolites 4. non-pivotal ED clinical studies 5. Non pivotal) Early GLP studies Regulatory validation Refer to EMA / FDA / cfda / MHLW Should be reserved for pivotal studies that require regulatory action for approval or labeling, such as BE or PK studies Room for more areas but not specified to date 27/11/2017 htt-sp:// 19

20 The focus of the recommendation Pre-GLP GLPà POC Post POC Screening/qualified accepted best practice Scientific validation Validation (or Regulatory Validation ) accepted best practices 27/11/2017 htt-sp:// 20

21 EBF recommendation - areas Five Six areas where applying scientific validation can be or has proven to be a valid alternative for regulatory validation. 1. All Urine analysis clinical/preclinical 2. All tissue homogenate analysis 3. All metabolites analysis prior to decision which metabolites require continued quantification using regulatory validation based on ICH(M3) and EMA-DDI: 4. A selection of non-pivotal ED clinical studies: focus on phase 0 and First into Man 5. All (Non pivotal) Early GLP studies: principles of scientific validation are compatible with GLP regulations 6. In Vitro studies 27/11/2017 htt-sp:// 21

22 Recommended approach Ø Not built on loose sand: Builds on 25 y of experience with Guideline criteria Refines them to the right balance of scientific requirements, scientific freedom and resource requirements 27/11/2017 htt-sp:// 22

23 How does it work Pre-study acceptance criteria In-study acceptance criteria if only in-study validation is performed, include (with predefined acceptance criteria) relevant missing parameters from pre-study scientific validation 27/11/2017 htt-sp:// 23

24 In detail: prestudy validation CoA with at minimum proof of identity/ purity Calibration curve: number of calibration samples Acceptance criteria CAL Matrix QC identical as study QC levels replicates 27/11/2017 Guideline Metabolites in plasma (ICH-M3) Urine Tissue homogenate assay appropriate scientific validation Early dev clinical studies Early dev preclinical studies stage appropriate scientific validation htt-sp:// 24 In Vitro Assays assay appropriate scientific validation y N Y or use dosed batch tbd Minimum 6, covering range incurred samples 75% and at least 5 points within 15% LLOQ) Minimum 5, covering range incurred samples 75% and at least 5 points within 20% LLOQ) Minimum 6, covering range incurred samples 75% and at least 6 points 75% and at least 5 points within 20% within 25% LLOQ LLOQ) y Y Y (or matrix matching) Y tbd 4 (LLOQ/low/ mid/high) min. 5 reps 3 (low/mid/high) min 3 reps 4 (LLOQ/low/mid/high) min. 5 reps tbd tbd

25 Finally vs Is the anxiety around more liberal acceptance criteria a valid worry or are we over-reacting? For SMOL BA current practice = , but most studies come in significantly better. In practice, also with more liberal criteria, most studies will be coming in at better acc&prec, than , and likely even Having more pre-agreed liberal acceptance criteria, will prevent undue repeat for those studies which fall outside (too) stringent criteria for the purpose of the study 27/11/2017 htt-sp:// 25

26 Proposed Strategy EBF Meeting 14 Nov 2017 Plasma Protein Binding Team put together to put forward approach for the 3 area s of PPB Early Development In vitro comparisons pre IND Ex vivo clinical comparisons 26

27 CoA with at minimum proof of identity/ purity Calibration curve: number of calibration samples Acceptance criteria CAL Matrix QC identical as study QC levels replicates 27/11/2017 Guideline Metabolites in plasma (ICH-M3) Urine Tissue homogenate assay appropriate scientific validation Early dev clinical studies Early dev preclinical studies stage appropriate scientific validation htt-sp:// 27 Plasma Protein Binding Stage 1 assay appropriate scientific validation Y N Y or use dosed batch tbd Minimum 6, covering range incurred samples 75% and at least 5 points within 15% LLOQ) Minimum 5, covering range incurred samples 75% and at least 5 points within 20% LLOQ) Minimum 6, covering range incurred samples 75% and at least 6 points 75% and at least 5 points within 20% within 25% LLOQ LLOQ) y Y Y (or matrix matching) Y tbd 4 (LLOQ/low/ mid/high) min. 5 reps 3 (low/mid/high) min 3 reps 4 (LLOQ/low/mid/high) min. 5 reps tbd tbd

28 Future Plans Ø EBF recommendations Plasma Protein Binding Ø Possible topics herg in vitro metabolism 28

29 Case Study The Exception! Ø Butenafine antifungal agent in Lotrimin Ultra cream Ø Current Formulation 1% w/w butenafine HCl with 0.3% w/w diethanolamine (DEA) Ø New Formulation 1% w/w butenafine HCl with 0.43% w/w triethanolamine (TEA)

30 Bioanalytical Procedure Ø Ø Ø Develop & Validate Analytical Method to determine concentration of test item(s) in:- 1. Receptor Fluid 2. Skin Wash 3. Tissue Swabs 4. Stratum Corneum (Tape Strips) 5. Skin 6. Formulation Undertake Pilot Study Undertake Analysis of Current and New Formulation

31 Methods Ø Place skin into cell Ø Apply formulation Ø Collect receptor fluid over time Ø Wash skin with soap and paper tissue after a representative period of time Ø Terminate Skin layers o Stratum corneum removed by tape stripping o Epidermis o Dermis Ø Analyse by LC-MS/MS

32 Comparison between Current and New Formulation of Butenafine Hydrochloride New Formulation % Applied dose (mean ± SD) Dislodgeable dose 24 h ± ± 6.40 Stratum Corneum (Tapes 1-5) 2.58 ± ± 1.68 Stratum Corneum (Tapes 6-10) 0.92 ± ± 0.32 Stratum Corneum (Tapes 11-15) 0.59 ± ± 0.25 Stratum Corneum (Tapes 15-20) 0.31 ± ± 0.09 Stratum Corneum (Total) 4.40 ± ± 2.16 Total unabsorbed ± ± 5.92 Total absorbed (penetrated) 0.04 ± ± 0.09 Dermal delivery 1.71 ± ± 1.43 Mass balance 92.2 ± ± 5.84 Current Formulation % Applied dose (mean ± SD)

33 Case Study Summary Ø Replacement of DEA in current formulation with TEA in proposed new formulation did not affect the extent to which butenafine HCl was absorbed or distributed through human skin Ø The in vitro data demonstrated that the clinical performance of the 2 formulations are indistinguishable Ø Data accepted by FDA and resulted in authorisation to market of new Lotrimin Ultra cream Ø In vitro skin permeation studies can be a suitable surrogate for more expensive and lengthy bioequivalence studies

34 Thank You for your Attention Many thanks to my EBF colleagues who were involved in the mws so far and ones to come 34

35 References ICH guideline S2(R1) Guidance on Genotoxicity Testing and Data Interpretation for Pharmaceuticals Intended for Human Use November 2011 ICH guideline M3(R2) on non-clinical safety studies for the conduct of human clinical trials and marketing authorisation for pharmaceuticals December 2009 ICH S7A Safety Pharmacology Studies for Human Pharmaceuticals November 2000 ICH S7B Nonclinical Evaluation of the Potential for Delayed Ventricular Repolarization (QT Interval Prolongation) by Human Pharmaceuticals. May

36 References EMA Guideline on Bioanalytical Method Validation July 2011 OECD General Questions and Answers Concerning OECD Principles of Good Laboratory Practice (GLP) and Mutual Acceptance of Data (MAD) November 2015 Tiered Approach into Practice: Scientific Validation for Chromatography- Based Assays in Early Development a recommendation from the European Bioanalysis Forum. Bioanalysis (2015) 7(19), Use of an in vitro Human Skin Permeation Assay to Assess Bioequivalence of Two Topical Cream Formulations Containing Butenafine Hydrochloride (1%, w,w). Regulatory Toxicology and Pharmacology 82 (2016)