S. Di Giovine Business Development Director 29 Novembre 2011 Università degli Studi di Milano Via Golgi 19 aula G10

Corporate profile

TOP srl, originated in 2006 as a spin-off of the University of Milan and has been backed in 2007 by the NEXT fund managed by Finlombarda Gestioni SGR.

TOP is located at the Parco Tecnologico Padano in Lodi, near Milan.

TOP research team is specialized in the generation of vectors, cells and animals for Molecular Imaging using:  Bioluminescence  PET (Positron Emission Tomography)  NMR (Nuclear Magnetic Resonance)

TOP mission

To provide the scientific community with non-invasive tools for the study of the activity of drugs and environmental or food contaminants TOP aims at combining the power of physics, informatics and molecular biology to generate novel models for the bio-medical research and for the control of the environment and alimentary chain.

TOP strives to renew present investigational tools in the respect of animal rights to satisfy two conflicting needs: use of predictive biological models in the absence of animal distress.

The Company expertise: Molecular Imaging

Molecular imaging is based on the appropriate use of REPORTERS: molecules easily detectable and allowing to study spatio-temporally molecular events in living organisms.

NR NR NR Reporter green fluorescent dye

The Company unique and proprietary technology*: the use of insulator sequences to obtain generalized expression of the reporter CCD Camera

* EU patent No. EP 1298988B1 US patent No. 7,943,815

All TOP products are subjected to intense validation to ensure:

PREVENTION OF EPIGENETIC MODIFICATIONS REPRODUCIBLE and REGULATED EXPRESSION OF THE REPORTER REPORTER EXPRESSION MAINTAINED IN TIME AND GENERATIONS

SATURN

VALIDATION

constitutive/ inducible S PECIFICITY A CCURACY T URN-OVER R EPRODUCIBILITY U BIQUITY N ON-INVASIVENESS

TOP products enable to study your favorite molecular process along the whole life cycle E 19.5

P.N. 1 E 18.5

E 17.5

16.5

E 16.5

15.5

E 14.5

DAY 14.5

14.5

E 12.5

Embryonic Dev. & Pregnancy 18.5

P.N. 10

3 months

Adulthood

9

TOP finds a response to a variety of research needs



Pharma:

− identification of novel drug targets − speeding of pre-clinical testing − identification of drugs endowed of tissue specificity of action − more predictable toxicological tests − gender pharmacology − pharmacology of infancy and aging − new indications for drugs in the clinic or in the market 

Environmental & Food toxicology:

− identification of specific activities in mixtures of compounds − threshold effects in short and long term exposures − − identification of food contaminants threshold effects in short and long term exposures − identification of the organs targeted by environmental pollutants 

Biomedical research:

− understanding the physiological effects of specific molecular events

One single tool for the multidimensional analysis of drugs in vivo.

 Immediate readout;  Increased prediction power: - holistic vs. reductionist approach; - drug action, not only distribution.

 Increased ability to perform multidimensional analysis of drug activity: - space (organs, PD); - time (PK, longitudinal studies); - immediate evidence of undesired activity (toxicology).

TOP Reporter system

 Lower costs;  Decreased research time;  Significantly reduced number of animals.

Partnership

TOP srl is partner of Caliper Life Sciences (former Xenogen) , the world leader for in vivo optical imaging.

TOP reporter models - I

 repTOP™ mice available at present:

Models repTOP™ ERE-Luc mouse

(Estrogen Receptors)

repTOP™ PPRE-Luc mouse

(PPAR’s)

repTOP™ MITO mouse (“MITO as in MITOSIS”)

repTOPTM Ubi-Luc2 (series) mouse + VivoGlo TM

Therapeutic areas:

• Aging-related pathologies • Depression • Inflammation • Lipid metabolism alteration • Diabetes • Cancers and others • Oncology • Regenerative medicine • Toxicology • • Metabolism (CYP3A4) Apoptosis (Caspase 3/7)

TOP reporter models - II



repTOP™ mice soon available

Models

repTOP TM

NFkB-Luc2 mouse

repTOP TM

TRAIL-EGFPLuc mouse

repTOP TM P21-Luc /EGFPTK mouse repTOP TM ARE Luc2

Therapeutic areas:

Apoptosis Inflammation Proliferation Oxidative stress In addition, TOP has generated a vast array of constructs for cell and animal engineering ready to be used and addressing most of current major needs in biological research. All of these constructs contain two or more reporters enabling bimodal and trimodal applications of molecular imaging

TOP reporter models - III

Custom- made repTOP™ mice: TOP’s technology platform and the standardized validation process are the guarantee of a rapid and reliable generation of an industry standard-compliant reporter system.

Feasibility Generation Characterization Validation 4-5 weeks 15 -18 weeks Founder mouse

OUTSOURCING YOUR RESEARCH AT TOP

The expertise of TOP research team in molecular imaging is at our client’s service for advise and testing of compounds in a wide variety of therapeutic areas

TOP mice in the world

 Italy: − − Spedali Civili, Brescia Roma, University Tor Vergata  Europe: − Institut de Genetique et de Biologie Moleculaire et Cellulare (IGBMC)  USA: − University of Kansas Medical Center (KUMC) − University of Miami, University of Chicago − University of Illinois − − UT Southwestern Medical Center at Dallas University of Pennsylvania School of Medicine − Karolinska Institute − University of Tennessee − Max Planck Institut fuer Psychiatrie Muenchen − Dana-Faber Cancer Institute − Dartmouth Medical School − Baylor College of Medicine.

− Leiden University Medical Center.

 Australia: − Prince Henry’s Institute of Medical Research  Leading Pharmaceutical and Biotechnology companies worldwide.

TOP newest product: repTOP TM mitoIRE to study proliferation in vivo

The repTOP

TM

mitoIRE mouse to study

proliferation in vivo

Anti – proliferative stimuli Pro–proliferative stimuli 1 u

NF-Y promoter CCD Cam era Cyclin B2 minimal promoter

repTOP

TM

mitoIRE validation

cell proliferation induced by application of 7,12-dimethylbenz(a)anthracene to the skin of the left ear can be easily measured in vivo

150000 100000 50000 0 0d 1d 2d

Time (days)

3d 6d 9d

COMPARING the sensitivity of Ki-67 immunohistochemistry and bioluminescence in the MITO mouse Ki-67 immunoistochemistry d3 d9 d3 Imaging d9 Bioluminescence: low background and rapid detection 0,3 0,2 0,1 0 d3 d9 80000 60000 40000 20000 0 d3 d9

repTOP TM mitoIRE following in time the development of chemically induce prostate carcinoma 200 100 12 4 1 0

Combining repTOP TM and VivoGlo TM technologies repTOP™ Ubi Luc2-M 8 mouse Z-DEVD-Aminoluciferin, Sodium Salt Induction of liver apoptosis 6h i.p. treatment with: D-GalN (800mg/kg) + LPS (100 µg/kg )

Nakama T et al. Hepatology. 2001