The immune response CRO Founded in 2016 Location BioHub at Alderley Park (UK) Management Team Dr. Ir. Robert-Jan Lamers Dr. Masih Alam

OUR BUSINESS A CRO offering specialised lab services in the field of immunology: General immune and bio-analysis of drugs in discovery and research Immunogenicity analysis - preclinical in vitro human immune analysis of drugs TARGET CLIENT GROUPS Drug Discovery Companies Research Institutions CROs for drug analysis

General immune analysis Immunity assays Cytokines, growth factors or antigen-specific detection Cell mediated immunity FACS, ELISA Molecular assays Histopathology Microscopy and imaging Animal models and animal based immunology Immune Modelling – experimental designs Mass spectrometry

SCOPE – Immunogenicity of drugs Translation from preclinical (animal) studies to humans (clinical trials) needs urgent improvement >> demands preclinical human systems Immune response (immunogenicity) can be caused by a drug but also by drug-induced tissue damage Any drug molecule has the potential to cause unwanted tissue damage Current immunological risk (immunogenicity) assessment does not include immune response against tissue damage, causing unsafe drugs being tested in clinical studies or even being used in treatment This is a gap in the market! This is the scope for novel in vitro human immune analytical systems.

Drug Induced Secondary Disease Common but currently outweighed due to benefits of ‘risk’ drugs: Neuropathy (idiopathic neuropathy) Interstitial Lung Disease (DI-ILD) Vasculitis Drug Induced Lupus (DIL) Immune Thrombocytopaenia Immune based demyelination Myopathy >300 drugs are reported to cause DI-ILD (example) >2 million cases of adverse drug reactions in the US annually including 100,000 deaths 250,000 cases of hospital admissions in the UK annually €430M could be saved in the Netherlands if side effects are reduced

Our approach: COMPIT – A NOVEL UNIQUE SYSTEM In vitro human immune model Cell culture systems adapted to test drug-induced damage Focused on novel biomarkers for immune response due to (drug- induced) tissue damage using cutting edge analytics (patent foreseen) Incorporates mathematical system to score immunological risk and predict secondary pathology (patent foreseen) Fits in preclinical studies

Preclinical studies: where does COMPIT fit? In vitro studies In vivo studies COMPIT COMPIT General Characterisation Immune Response against drug and drug-induced tissue damage N O A E L Toxicological studies Pharmacological studies Mid Dose M T D Receptor characterisation Binding assays Enzyme inhibition Cytotoxic activity Immunological Safety Safety Efficacy Immunological Safety (Potential) Inhibition of Danger Signals Dose conversion

Fields of application COMPIT can be applied to Small molecules Large molecules Generics and biosimilars Potentially predictive (preclinical) of ADA development Particularly the research and development of large molecules is evolving and expanding worldwide >> requiring more immune response analysis Regulatory Bodies increasingly seeking/advising analysis of safety based on new immune markers – these are included in COMPIT

Diffuse Alveolar Damage Example for drug induced interstitial lung disease Non Target Tissue Tissue Damage Pathology Disease Amiodarone Target tissue cardiac but …….. ROS – oxidative stress Metabolites – interference with tissue Modification of cell biochemistry e.g. covalent binding ATP depletion or Loss of Ca++ homeostasis Damaged mitochondria – increased cell permeability PNEUMOCYTES Epithelial cells via Inhalation or Circulation Inflammation Alveolar and Bronchial area Dysfunction/Death of Cells Apoptosis to Necrosis Compromised Alveolar Wall Hyaline + Protein rich Exudate in Alveolar Space Diffuse Alveolar Damage or Organising Pneumonia Hyaline FIBRIN Reactive epithelial cells Danger mediated antigen processing PROGRESSIVE IMMUNE RESPONSE Scarring Immune mediated damage + phagocytosis Maturation and immune response generated More examples: Bleomycin Amphotericin B Rituximab Nitrofurantoin Lung Fibrosis DI ILD COMPIT ANALYSIS OF DAMAGE AND IMMUNOGENICITY

Benefits – Unique Buying Points Better translation from preclinical studies to humans Reduction of the number of animal tests A more robust and comprehensive immunologic profile: improved safety and risk prediction thus reducing human health hazard risk Drug development can be terminated before costly advanced stages of drug development: reduced investments in non-viable drugs and reduced attrition rate Potential to improve drugs and make non-viable drugs accessible to patients Meeting changes to expected regulatory requirements on safety

Why? FDA 2014 guideline for ‘Immunogenicity Assessment for Therapeutic Protein Products’ mentions issues related to tolerance to endogenous proteins and autoimmunity through immune response against endogenous proteins Latest version of EMEA guideline on immunogenicity assessment of biolotechnology derived therapeutic proteins mentions “while most effort is usually focused on antibody detection and characterisation as this is often related to clinical safety and efficacy, cell-mediated responses are also important and their assessment may be considered by applicants” EMEA meeting (January 2016) “…. a highly interesting and important platform for pharma and biotech companies that meets an urgent demand” Experienced in immunology and analytical chemistry Access to high-end facilities, professionals and network through location at BioHub campus

Projects for PhD placements You can work in our lab in the COMPIT project through 2017 and 2018 if your experience fits the requirement of the post 3 month placement period You will need to bring your own funding for living and travel Work location is Alderley Park near Macclesfield. You will be in a large campus where there are many other companies (mostly discovery and CROs involved in the drug industry)

Project 1: Tissue Modelling Purpose: Develop and maintain 3D tissue systems, organoids Use of hydrogels/matrigels and specialised tissue growing wells. 3 month duration Period Sep – Nov 2017 or any other time Location: BioHub, Alderley Park Skills & Learning: Cell culture, mammalian cells 2D and 3D systems, organoids Primary and established cells, Stem cells, media preparation Histology, microscopy and imaging

Project 2: Assay Development Purpose: To extract and measure immunogenic particles from damaged tissue in culture. Use of Mass Spec and immunologic assays for analysis. 3 month duration Period Sep – Nov 2017 or any time after that Location: BioHub, Alderley Park Skills & Learning: Cell culture Protein identification and quantification Western blotting, ELISA, MS, RNA methods Experimental designs

Project 3: Bio analysis Purpose: Skills & Learning: Assess the immunologic events that follow the tissue damage leading to an immune activation Use of biomolecular and immunologic assays for analysis. 3 month duration Period Jun – Aug 2018 Location: BioHub, Alderley Park Skills & Learning: Cell culture, tissue, lymphocytes Protein identification and quantification Western blotting, ELISA, Flow Cytometry, immunohistochemistry Experiment designs Statistics

Project 4: Cell Interactions Purpose: Design and maintain experiments that allow interaction of lymphocytes with damaged tissue Use of general and customised cell culture systems 3 month duration Period Sep-Nov 2017 and periods in 2018 Location: BioHub, Alderley Park Skills & Learning: Cell culture, tissue, lymphocytes Cell migrations 2D and 3D cells, organoids Microscopy and imaging Experiment designs

Project 5: Mathematical Model Purpose: Assimilating and analysing data from cell culture studies for determining immune response scores. This will be crucial to validating the COMPIT system for preclinical immune response analysis. Use of mathematical model designed by Chester University for translating experimental data. 3 month duration Period Jun – Aug 2018 Location: BioHub, Alderley Park Skills & Learning: Mathematical modelling in biological experiments Statistics Understanding of cell based biological experiments and cellular pathways. Experimental designs Overview of drug toxicology and preclinical studies

My Contact DR. MASIH ALAM IMMUNOSYS LTD IMMUNDNZ LTD E. masih.alam@immundnz.com, masih.alam@immunosysltd.com www.immunosysltd.com www.immundnz.com