1. 1.Areas where new development tools could accelerate progress -- Formulation -- Glycosylation 2.Potentially important future areas of medical development -- Nanotechnology -- Tissue Engineering

2. Impact of TransForm Technology TraditionalTransForm Ability to explore F&F space more effectively and efficiently solvent Other variables process  Minimal  1 – 2 Months      Deep & iterative  2 – 4 Weeks  200-20,000+  10-20 # of F & F experiments Timing Informatics; data mining, learning

4. Case Study: Ritonavir 1.5 years after launch, converted into unanticipated form II polymorph 50% less soluble Abbott compelled to recall & reformulate Form IForm IIForm III MPT 122 °CMPT 125 °CMPT 80 °CMPT 97 °C MPT 116 °C Form IVForm V Within weeks at TransForm, using < 2g: Both known forms identified & characterized Found three novel, previously unreported forms Novel, robust methods to make each form Morissette et al. PNAS 100, 2180 (2003).

5. New Tools 1.Imaging 2.Informatics 3.Genomics 4.Proteomics 5. Glycomics

6. “Cracking the Code” of Sugars is Analogous to the Sequencing of DNA  The sequencing of DNA has laid the foundation for biotechnology revolution  Like DNA and proteins, sugars play a central role in regulating basic biological activity, disease mechanisms, and drug action  Sugars exist as sequences of building blocks similar to DNA, but there has been a lack of adequate sequencing tools  Understanding of sugars is critical for polysaccharide drugs (e.g. Lovenox) and glycosylated proteins (e.g. Epogen) BNR 3 A G C T A G C O O O OH -OOC OSO 3 - CH 2 OSO 3 - OH NSO 3 - O O O OH OSO 3 - CH 2 OSO 3 - OH NHSO 3 - O COO - O O O OH O S O 3 - CH 2 OSO 3 - OH NHS O 3 - O COO - O O O OH 3 - O 3 - OH

7. Inherent complexity of sugars has prevented comprehensive understanding  Structural complexity and information density  Lack of amplification  Heterogeneity The Problem: Lack of technology to and tools to sequence sugars has made it difficult to characterize and engineer sugars, and decipher their role in biology.

8. Convergence on unique solution to complex sequences Sequencing Complex Polysaccharides [1999] Science 286: 537-542. Momenta Pharmaceuticals Mass signatures of groups MALDI -MS Mass of chain – chain length NMR CE Quantitative building block information Multiple Enzymes Linkage information Integration of Data ESI-MS

9. Future areas of medical development 1.Nanotechnology 2.Tissue Engineering

10. Prototype Device Silicon Nitride or Dioxide Cathode Active Substance Anode Silicon

11. Implantable Drug Delivery System Battery-powered, telemetry-controlled implant Design based on pacemaker and ICD microelectronics

12. Reservoir Opening Mechanism

13. Pre-Clinical Studies Demonstrate in vivo Release Experimental Protocol Implant microchips subcutaneously in rats Release radioactive mannitol ( 388 ng/well non-metabolized sugar) Collect urine and analyze for radioactive content as an indicator of drug release

14. Polymer Therapeutics : Nanosized medicines polymeric drug or sequestrant polymer-drug conjugates polymer-protein conjugate linker drug targeting residue polymeric micelle hydrophilic block hydrophiobic block drug polyplex polymer-DNA complex DNA cationic block hydrophilic block 40-60 nm 60-100 nm 5-15 nm Mw = 5 - 40,000 Da protein ~20nm

15. 1.How do you assess safety? 2.How do you characterize nanomedicines --Biological --Physical/chemical 3. What animal models are appropriate?

16. Annual Tissue Loss End Stage Organ Failure (U.S.)  Over $500 billion in health care costs  40 to 90 million hospital days  8 million surgical procedures

17. Incidence of Organ and Tissue Deficiencies Bone Joint replacement 558,000 Bone graft275,000 Internal fixation480,000 Facial reconstruction30,000 Cartilage Patella319,400 Meniscus250,000 Arthritis (Knee)149,900 Arthritis (Hip)219,300 Small Joints179,000 Tendon33,000 Ligament90,000 Skin Burns, Sores, 3,650,000 Ulcers1,100,000 Heart754,000 Blood Vessels606,000 Liver205,000 Pancreas728,000

18. In vitro Tissue Culture Biodegradable Polymer Scaffold Cells Osteoblasts Chondrocytes Hepatocytes Enterocytes Urothelial Cells In Vivo Implantation New Bone Cartilage Liver Intestine Ureter

20. Cartilage Tissue Engineering BEFORE cell seeding AFTER 2 weeks in culture

22. System  Modified PGA Tubes  8 Weeks SMC Culture, then EC  Bio-Reactors – Pulsatile Radial Stress

23. Medium Reservoir Flow Direction Pulsatile Pump Compliance Chamber Magnetic Stirplate 20 cm 4 Bioreactors, Assembled in parallel

25. Characteristics  50% Collagen  Rupture Strengths > 2000 mg Hg  Suture Retention – Strengths up to 90g  Demonstrates Contractile Responses to Serotonin, endothelin-1, and Prostaglandin F2α

27. PECAM1 CD34 Human Embryonic Endothelial Cells Form Functional Blood-Carrying Microvessels

30. 1.How should safety be assessed? 2.What are appropriate markers? 3.How do you determine appropriate function? 4.What are appropriate animal models?