1.areas where new development tools could accelerate progress -- formulation -- glycosylation...
TRANSCRIPT
1. Areas where new development tools could accelerate progress
-- Formulation
-- Glycosylation
2. Potentially important future areas of medical development
-- Nanotechnology
-- Tissue Engineering
Impact of TransForm TechnologyImpact of TransForm Technology
Traditional TransForm
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
Case Study: Ritonavir
• 1.5 years after launch, converted into unanticipated form II polymorph
• 50% less soluble
• Abbott compelled to recall & reformulate
Form I Form II Form III
MPT 122 °C MPT 125 °C MPT 80 °C MPT 97 °C MPT 116 °C
Form IV Form V
Within weeks at TransForm, using < 2g:• Both known forms identified & characterized
• Found three novel, previously unreported forms
• Novel, robust methods to make each formMorissette et al. PNAS 100, 2180 (2003).
New Tools
1. Imaging
2. Informatics
3. Genomics
4. Proteomics
5. Glycomics
“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)
B N R 3A G C T A G C
O
O
O
O H
- O O C
O S O3-
C H2
OS O3-
O H
N S O3-
O
O
O
O H
O S O3-
C H2
O S O3-
O H
N H S O3-
O
C O O-O
O
O
O H
O S O3-
C H2
O S O3-
O H
N H S O3-
O
C O O-O
O
O
O H
3-
O3-
O H
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.
Convergence on unique solution to complex sequences
Sequencing Complex Polysaccharides [1999] Science 286: 537-542. Momenta Pharmaceuticals
Mass signatures of groups
MALDI-MSMass of chain
– chain length
NMR
CEQuantitative
building block information
Multiple Enzymes
Linkage information
Integration of Data
O
O
O
OH
OSO3-
CH2OSO3-
OH
NHSO3-
OHCOO-
R
O
O
O
OH
-OOC
OH
CH2OSO3-
OH
NHAc
O
O
O
OH
OH
CH2OSO3-
OSO3-
NHSO3-
O
COO-
O
O
O
OH
OSO3-
CH2OSO3-
OH
NHSO3-
O
COO-
O
ESI-MS
Future areas of medical development
1. Nanotechnology
2. Tissue Engineering
Prototype DevicePrototype Device
Silicon Nitride or Dioxide
Cathode Active Substance Anode
Silicon
Implantable Drug Delivery SystemImplantable Drug Delivery System
Battery-powered, telemetry-controlled implant
Design based onpacemaker and ICDmicroelectronics
Reservoir Opening MechanismReservoir Opening Mechanism
Pre-Clinical Studies Demonstrate in vivo Release
time
curr
entExperimental 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
0
20
40
60
80
100
120
0 2 4 6 8 10 12 14 16 18
Time (hours)
Ma
ss
pe
r S
am
ple
(n
g)
0
400
800
1200
1600
2000
2400
Ma
ss
Re
co
ve
red
(n
g)
Polymer Therapeutics : Nanosized medicines Polymer Therapeutics : Nanosized medicines
polymeric drugor sequestrant
polymer-drug conjugates
polymer-proteinconjugate
linker
drug
targeting residue
polymeric micelle
hydrophilic block
hydrophiobic block
drug
polyplexpolymer-DNA complex
DNA
cationic block
hydrophilic block
40-60 nm
60-100 nm
5-15 nm
Mw = 5 - 40,000 Da
protein
~20nm
1. How do you assess safety?
2. How do you characterize nanomedicines
--Biological
--Physical/chemical
3. What animal models are appropriate?
Annual Tissue LossAnnual Tissue LossEnd Stage Organ Failure (U.S.)End Stage Organ Failure (U.S.)
Over $500 billion in health care costs
40 to 90 million hospital days
8 million surgical procedures
Incidence of Organ and Tissue DeficienciesIncidence of Organ and Tissue Deficiencies
Bone
Joint replacement 558,000
Bone graft 275,000
Internal fixation 480,000
Facial reconstruction 30,000
Cartilage
Patella 319,400
Meniscus 250,000
Arthritis (Knee) 149,900
Arthritis (Hip) 219,300
Small Joints 179,000
Tendon 33,000
Ligament 90,000
Skin
Burns, Sores, 3,650,000
Ulcers 1,100,000
Heart 754,000
Blood Vessels 606,000
Liver 205,000
Pancreas 728,000
In vitro Tissue Culture
Biodegradable Polymer Scaffold
CellsOsteoblastsChondrocytesHepatocytesEnterocytesUrothelial Cells
In Vivo Implantation
New
BoneCartilageLiverIntestineUreter
Cartilage Tissue EngineeringCartilage Tissue Engineering
BEFORE cell seeding
AFTER 2 weeks in culture
SystemSystem
Modified PGA Tubes
8 Weeks SMC Culture, then EC
Bio-Reactors – Pulsatile Radial Stress
MediumReservoir
Flow Direction
Pulsatile Pump
Compliance Chamber Magnetic Stirplate
20 cm
4 Bioreactors,Assembled in parallel
CharacteristicsCharacteristics
50% Collagen Rupture Strengths > 2000 mg Hg Suture Retention – Strengths up to 90g Demonstrates Contractile Responses to
Serotonin, endothelin-1, and Prostaglandin F2α
PECAM1
CD34
Human Embryonic Endothelial Cells Form Human Embryonic Endothelial Cells Form Functional Blood-Carrying MicrovesselsFunctional Blood-Carrying Microvessels
1. How should safety be assessed?
2. What are appropriate markers?
3. How do you determine appropriate function?
4. What are appropriate animal models?