An over view of the biopharmaceutical discovery process

1. An over view of the biopharmaceutical discovery process Robert Rehfuss, PhD

2. A Simplified Process Map for Biopharmaceutical Discovery Target Identification Laying the Groundwork Cranking up the Band Delivering the Goods 2-5 years

3. Step 1: Target Identification • A “Target” is the molecule (receptor, enzyme, etc.) that you want to activate/inhibit so as to effect a disease process • Choosing a target is the single most important step in the process • Good targets do not always result in therapeutic agents • Targets that don’t effect the disease biology cannot result in effective therapeutic agent ever, regardless of time/resources committed to project • Requires extensive and detailed knowledge of: • Specific disease area biology including pathophysiology • Existing therapy for a disease including pros/cons of current therapies • Current scientific literature directly (but also indirectly) related to disease • Most targets are widely known and well picked over • Capitalizing on new science (new targets, new approaches) requires speed to gain an advantage over competition

4. Leading Causes of Death for All Males and Females United States: 2003* Deaths in Thousands A Total CVD B Cancer C Accidents D Chronic Lower Respiratory Diseases E Diabetes Mellitus F Alzheimer’s Disease Source: CDC/NCHS and NHLBI. *Preliminary

5. Stable Atherosclerotic Lesion Fibrous Cap

6. Thrombus Formation and ACS Plaque Disruption/Fissure/Erosion Thrombus Formation Old Terminology: UA NQMI STE-MI New Terminology: Non-ST-Segment Elevation Acute Coronary Syndrome (ACS) ST-Segment Elevation Acute Coronary Syndrome (ACS)

7. Key Role of Platelet Aggregation in Arterial Thrombus Gachet , C. (2006) Ann. Rev. Pharmacol.Toxicol.

8. Trends in Age-Adjusted Prevalence of Health Conditions, U.S. Adults Ages 20-74 NHANES:1971-74 to 1999-2000 Source: Ann. Rev. of Nut. July 2004. Vol.24: 401-431

9. Biology beats Marketing every time!An innovative and effective cure for a rare disease will make more money than still another anti-hypertensive

10. Step 2: Laying the Groundwork • After choosing a target, two key strategic questions need answers: • Is the available data supporting the target sufficient to convince others to move a potential agent into ($$$) clinical trials? • Usually not. What additional studies need to be preformed? Who does them? What reagents (and from what source) are needed to enable those studies? What animal models will be used to demonstrate efficacy of the final therapeutic agent for the FDA? • What type drug will be most likely to produce both a clinical candidate and a profitable therapeutic agent? • Pros/Cons of biopharmaceutical vs. pharmaceutical approaches • Delivery Route, Cost of Goods, Likelihood of Success • Choice effects downstream strategy as different types of assays need to be developed to screen candidates • Recombinant production of target as either antigen or reagent for assays • Development of primary assay (ELISA vs. Binding vs. other)

11. Step 2: Laying the Groundwork (cont’d) • And one tactical question: • How will the activity of potential candidates be measured? • Binding or Enzyme assays? • Cell based assays? • Where do the reagents come from? • Who designs the assays? Who performs the assay(s)?

12. Monoclonal Antibody Generation Recombinant production required 100’s of mAb to screen

13. ELISA Recombinant production required

14. Step 3: Cranking up the Band(BioPharmaceuticals) • Assembling a team of scientists to sort through the different molecules to find the specific one with the right mixture of properties • Affinity for Target • ELISA, HTRF, SPR (Biacore), BLI (Octet RED) • Ability and potency to inhibit desired biology • Cell or plasma based signaling assays • Pharmacokinetics/ Pharmacodynamics (PK/PD) • Other Biophysical Properties • Expression yield (system dependent) • Melting temperature and behavior • Low temperature, irreversible melting behavior very undesirable • Aggregate formation on storage Properties can be “tweaked” by addition of point mutations at this step

15. Step 3: Cranking up the Band(Pharmaceutical) • Assembling a team of scientists to sort through the different molecules to find the specific one with the right mixture of properties • Affinity for Target • ELISA, HTRF, SPR (Biacore), BLI (Octet RED) • Ability and potency to inhibit desired biology • Cell or plasma based signaling assays • Pharmacokinetics/ Pharmacodynamics (PK/PD) • Optimization of dosing formulations • Other Biophysical Properties • Solubility, Crystalline Behavior, Stability • Key parameters for manufacturing Properties “tweaked” by subtle variation of chemical structure

16. Generic Triage Scheme New compound synthesis by Chemistry Tested in Primary Assays for target potency/selectivity Doesn’t meet criteria • Tested in Secondary Assays for potential in vivo activity • Screening assays for target activity • Screening assays for PK potential • Screening assays for potential liabilities • In vivo testing : • PK/PD (ADME) • Efficacy (Biology) & • Toxicity Assessment Candidate declaration

17. Step 4: Delivering the Goods • Complete characterization of a pre-clinical candidate in non-human species. Documentation produced with an eye on the data required by FDA for filing of IND/BLA • Production of material under GMP (or sometimes near-GMP) conditions • Extensive testing in disease models to demonstrate likelihood of clinical efficacy • If mAb doesn’t cross react with non-human model then 2 completely separate programs needed • Extensive PK/PD testing in multiple species to try and anticipate human PK/PD • Toxicology studies (high dose and prolonged exposure in at least 2 established species (rat, mouse, dog, NHP) one of which may be rodent • Final determination of expression system and protocol to be used for GMP production • FDA will only accept data for IND/BLA if material is GMP and studies are carried out under GLP conditions. • Studies with near-GMP material need replication if they are to be reported

18. Questions?