1. Role of CMD Animal Models: What is good and what are we missing? Markus A. Rüegg Biozentrum University of Basel, Switzerland

2. What do we need animal experiments for? Provide GO/NO-GO decision for sponsor to advance a new treatment approach into clinical trials (i.e. factor of risk management) Generate proof of concept and in vivo efficacy data for regulatory authorities in preparation to “first in man” studies Provide information on effective dose, route of administration Allow ranking of potential drug candidates according to efficacy (head to head comparison; evidence for combination therapy) Support estimates of effect size to be expected in clinical trials (Generate toxicology, safety pharmacology & PK data ) In an ideal setting we expect answers to:

3. What are the levels of animal experimentation? Understanding the disease: molecular level cellular level organ level Discovery of potential drug targets Validation of potential drug targets Treatment discovery Formal Requirements $ $ $$ $$$$ Costs academia industry +++++ +++++ +++++ Sponsor Each step should allow to make GO/NO-GO decisions for the next step

4. Efficacy data in animals- what is required by regulatory authorities? Example: EMA’s guidance document for non-clinical part of a critical assessment report: “… mode of action and brief rationale for the development of the product in the proposed indication should be provided” Report on primary pharmacodynamics shall include documentation on:  Proof of concept (in vitro and in vivo) and mode of action  Availability of animal models relevant to the proposed indication Unlike for toxicology studies, there are limited guidelines on the type of efficacy studies needed to support IND filing for a new therapeutic approach

5. What animal models should be used ? Objectives: Provide efficacy data to support first in man study Increase predictability of clinical outcome based on animal data Challenges & Bottlenecks: Selection of species:rodent mammal (mouse) non-rodent mammal (e.g. dog) non mammal (fly, fish) Selection of genetic/strain background Selection of type of mutation:“genotypic match” “phenotypic match” For genetic diseases, the genotypic match of the animal model is highly desirable; a phenotypic match would be ideal

6. Issues arising using animal experiments 1: Jim Giles. Nature (2006), 444; 981 2: Perel et al. (2006) Br. Med. J. Dec 2006, 1-16 “Small scale studies are pointless if they do not produce results people have confidence in” 1 Animal studies frequently poorly designed as they often are:  Underpowered  Unblinded  Not randomized “Because animal experiments are part of the evidence used to decide which interventions are taken forward in clinical trials, efforts to avoid bias and random error are as important … for animal models as … for clinical trials. 2 “Animal experiments under fire for poor design” 1 The proper conduction of animal experiments to test for efficacy is independent of the disease investigated and is key to provide the rationale for clinical trials

7. What is needed for effective treatment development? Definition of effective models for drug screening (cell models, non mammalian systems) Recommendations for use of mammalian models for efficacy studies Consensus for disease-relevant and predictive animal model(s) Consensus for biochemical, histological and functional assays Standardization of protocols (timing of drug application,…) Recommendations to regulatory authorities Specific objectives of TREAT-NMD for DMD and SMA (kick off in 2007)

8. Brussels 17-19 November 2009 Efforts to create SOPs for animal models of DMD Joint effort of several groups worldwide Workshops held in Washington DC and Zurich Consensus on animal models Consensus on read-outs Setting up of working groups to establish SOPs SOPs published and disseminated (autumn 2008) http://www.treat- nmd.eu/research/preclinical/SOPs/ approximately 700 downloads/month http://www.treat- nmd.eu/research/preclinical/SOPs/

9. Brussels 17-19 November 2009 Efforts to create SOPs for animal models of DMD Evaluation of questionnaire

10. Brussels 17-19 November 2009 Efforts to create SOPs for animal models of DMD Which readout do you routinely use? Histology:100% Molecular analysis:81% Serum enzymes: 74% Muscle strength in vivo:70% … Imaging (MRI; Echo):20% Electrophysiology:10%

11. Brussels 17-19 November 2009 Efforts to create SOPs for animal models of DMD Satisfaction and awareness 85% know the SOPs 50% use the SOPs > 90% would use SOPs if setting-up a new lab > 90% think that the SOPs help foster faster track to treatment

12. Brussels 17-19 November 2009 Efforts to create SOPs for animal models of DMD New issues to be addressed in DMD: Should there be a platform to publish negative results? Enthusiastically received (> 90% thought that this is a great idea)

13. What is the situation in CMD? Animal models: Heterogeneous set of diseases As many genetic mutations are known in humans, mice are probably the best species to model a particular disease subtype Subgroups: 1.abnormalities of α-dystroglycan glycosylation and defects in other membrane receptors (fukutin, POMGnT1, POMT1, POMT2, FKRP, LARGE, ITGA7) 2.abnormalities of extracellular matrix proteins (LAMA2, COL6A1, COL6A2, COL6A3) 3.abnormalities of nuclear proteins (Lamin A/C and nesprin) 4.abnormalities at the level of the endoplasmic reticulum (SEPN1). Additional mouse models need to be developed needs more available needs more

14. What is the situation in CMD? Experimental protocols: Many protocols developed for DMD can be adapted for CMD Development of a site where these protocols can be deposited and updated (TREAT-NMD?) There is a clear need for a person who coordinates and “pushes” such project forward