Alexeï Grinbaum_What is responsible about responsible innovation?
Michael Buschmann_Nanomedecine
1. Nanomedicine
Michael D. Buschmann
Department of Chemical Engineering
Institute of Biomedical Engineering
Ecole Polytechnique, Montreal, Canada
THE RESPONSIBLE DEVELOPMENT OF NANOTECHNOLOGY
CHALLENGES AND PERSPECTIVES
Ne3LS Network International Conference
2012 Nov 1, 2012
2. Overview
► Nanomedicine Products
► Why Nano for Medicine ?
► Requirements for Successful R&D
► New Technologies from Ecole Polytechnique
► Conclusion & Perspectives
3. Nanomedicine definition*
Nanomedicine uses nano-sized tools for the
diagnosis, prevention and treatment of disease
and to gain increased understanding of the complex
underlying patho-physiology of disease.
The ultimate goal is improved quality-of-life.
*European Science Foundation’s Forward Look Nanomedicine, 2005
“Nanobiotechnology” is a broader concept : fundamental cellular
mechanisms, molecular forces, molecular motors, cellular
electrochemical phenomena in nonhuman, plant and animal models.
4. Nanomedicine Products
•Iron Oxide or Gd Imaging Agents
Constrast agents for MRI
•Iron Oxide Supplements
To treat anemia
•Drug Nano Crystals
Increase oral bioavailability
•Liposomes Layered Micelle Nanoparticle
Drug delivery containers
•Polymer Therpauetics
Prolong and target action
•Nanoparticles
Multifuncitonal delivery systems
7. Marketed Nanomedicines Duncan Molecular Pharmaceutics 2011
Several marketed for > 20 years
8. Application Domains of Nanomedicine
Therapeutics
– Nanovectors for drug delivery – small molecules, proteins,
antibodies, DNA, RNA
– Targeted smart systems – hit the right tissue, the right cell
– Next generation medicines – RNAi
– Market > $50 billion
Diagnostics
– Imaging & contrast agents – magnetic, metallic, fluorescent
– Molecular sensors – arrays, chips, SPR etc
– Microfluidics – separation and analyses on a chip
– Personalized Medicine – make the treatment fit patient symptoms
and molecular profiling by diagnostics – emerging area
– Market > $5 billion
9. Point of Care Diagnostics - Target Applications
Ref: POC 2010 March 2010, Yole Développement SA.
10. Nanobiotechnology Market & Application Areas
(Includes Recombinant Proteins/Antibodies)
Ref: Genetic Engineering and Biotechnology News Oct. 15, 2010.
11. Why NANO ? Conner Nature 2003
Internalisation to Cells is Nano-size Dependent
12. Why NANO ?
Size controls
Routes of Drug Administration Biodistribution
Gaumet E J Pharm Biopharm 2008
13. Why NANO ? Gaumet E J Pharm Biopharm 2008
Endothelial Fenestration
Fenestration space is
Critical to
Nanosized : 1-1000nm
Biodistribution
Desirable size range 50-200nm avoids rapid
renal excretion and avoids liver and spleen
15. Dobrovolskaia Nanoletters 2008
Why NANO ?
Surface chemistry (PEG) determines protein
binding (opsonisation) and phagocytosis by
macrophages versus delivery to the target
19. Successful Research & Development
Requires
•Efficacy of Treatment
•Safety of Treatment
•Manufacturing Capability
•Cost-effectiveness
•Competitive Advantage
•Patent Protection
•Regulatory Approval
•Reimbursement Strategy
20. Safety – Cell-based Toxicity
•Cell Viability, Proliferation, Metabolism
MTT (cellular respiration) and related
•Cell membrane damage and necrosis
LDH release, direct membrane perturbation
•Apoptosis (programmed cell death)
Annexin V (early), TUNEL (late)
•DNA damage and genotoxocity
Strand breaks by COMET
•Oxidative Stress
O and N radicals damage multiple cell components
•Gene screening
Elucidate networks related to toxic responses
23. Groupe de recherche en sciences et
technologies biomédicales (GRSTB du FRQ-
S)
Chercheurs réguliers 30
Chercheurs-boursiers 11
Chercheurs associés 20
Étudiants (MScA & PhD) 183
Stagiaires post-doc 32
24. Regroupements de recherche du
FRQ
GRSTB
Biomatériaux Imagerie
& &
Biomécanique Traitement de
signal
Transfert
Transfert
de
technologique Nano-médecine
& connaissanc
aux Médecine
régénératrice es à la
entreprises
clinique
Pôle sectoriel
SGV
Plans stratégiques universitaires
25. Nanoparticles for Gene Delivery
(Buschmann)
Viral Vectors: Nonviral Vectors:
Retrovirus, adenovirus... Cationic lipids
High efficiency Cationic polymers (polycations)
Toxicity Greater safety
Immunogenicity Low efficiency
• Cationic polymers → low toxicity and ease of preparation
(ex: chitosan, polyethyleimine, poly-L-lysine, dendrimers..)
Intracellular
Condense DNA into transport Nuclear
3 localization
nanoparticles 4
1
Need strategies to Binding
to target
increase cells
2
Uptake into
cell 5
DNA release
transfection efficiency endosomes Gene expression 25
26. >1500 Clinical Trials in Gene Therapy
“FDA has not yet approved any human gene therapy product for sale.”
“EU has approved Glybera in 2012 for lipoprotein lipase (LPL)
deficiency in patients with severe or multiple pancreatitis attacks.”
27. Chitosan
• A frequently used natural polymer for gene delivery
• Biodegradable, biocompatible, non-toxic
OH OH
Chitin O O
O
HO HO
NHAc NHAc
Deacetylation
OH OH
O O
Chitosan O
HO HO
NH 2 NH2 +
DDA NH3 NHAc
NH 2 NHAc
Linear chain of glucosamine and N-acetyl-glucosamine
units linked by (1,4)-glycosidic bonds
We have previously developed a chitosan-based biomaterial to repair cartilage :
April 11 2012 : “Piramal Healthcare announces approval of its first
innovative product for regenerative medicine, BST-CarGel®”
28. Formation of DNA-chitosan complexes
Protonated amine Phosphate groups
groups (negative charges)
(positive +
charges)
+
- - Plasmid DNA
+ + -
+ -
+ + + +
+ + + - -
+
Chitosan - -
(in excess) -
-
DNA condensed into
Important factors: nanoparticles
• Chitosan molecular + +
+
weight and DDA +
+
• pH +
• N/P ratio (chitosan +
+
amine to DNA DNA/Chitosan
phosphate ratio) complexes
28
29. Influence of MW & DDA on Transfection
• Transfection efficiency as good or higher then state-of-the-a
• Strong chitosan structural –dependence of efficiency
30. Influence of MW & DDA on Transfection
* *
Transfection efficiency as good or higher then state-of-the-art
31. In vivo Delivery of Growth Factors FGF2 &
PDGF
M 1 2 3
pVax-1 ( 3kb)
4sFGF-2 ( 470 bp)
PDGFbb ( 327bp)
pVax1 vector
HindIII 4sFGF-24sFGF -2
or PDGFbb XhoI
PDGFbb
-
pVax1/4sFGF-2 (3 465 pb)
ou
pVax1/PDGF-bb (3 327 pb)
32. 4sFGF-2 Recombinant Protein and Anti-4sFGF-2
Antibodies in plasma following subcutaneous injection
of chitosan/pVax-4sFGF-2 nanocomplexes
92-10-5 increases protein
and reduces antibody
80-10-10 reduces protein
and increases antibody
33. Chitosan Properties Significantly
Modulate Protein and Antibody Levels
• Higher DDA and lower MW (92%DDA-10kDa)
efficiency expresses protein, minimises inflammatory
cell recruitment, lowers antibody response and
generates systemic circulating recombinant protein
• Lower DDA (80%DDA) augments inflammatory cell
recruitment and antibody levels, reducing circulating
protein, possibly useful for DNA vaccines
36. Maximal GLP-1 Expression with 92-10-
5
92-10-5 is the most efficient formulation
in both routes of administration
80-10-10 also effective
80-80-5 ineffective due to
immunostimulatory and poor
release properties as
demonstrated in the PDGF/FGF
study
GLP-1 expression is maintained
for 24 days following end of
treatment
greatly exceeding the lifetime
of current daily injected GLP-1
peptides
37. Glucose Tolerance Normalised with 92-10-
5
• Formulation 92-10-5
is the most effective
• Rapid return to
normoglycemia within normoglycemia
two hours
• No significant difference
between SC and IM
routes of administration
38. Nanoparticle Nucleic Acid Delivery
• siRNA delivery in Atherosclerosis (ApoB) and Cancer
(Helicase)
– Cell and animal studies
• Arthritis and inflammation
– pDNA/chitosan delivery of TNFα inhibitors (Etanercept , Enbrel)
• Manufacturing and Scale-up
– High volume mixing technologies, concentration, freeze-dry
– Transfer to cGMP facility with Quality Control methods
Freeze-dried, Room Temperature Stable
Pharmaceutical Product
39. Gold Nanoshell Probes for Atherosclerosis
Diagnosis and Personalized Medicine
Professor Fréderic Lesage
Optical absorbance at NIR
wavelength
-Easily tunable
-Monodisperse
-Non-toxic 39
42. No visible sign of toxicity after
injections of large doses
A Kidney Heart Spleen Liver B Saline control AuNS (OD=200)
Liver
Control
Saline
(OD=200)
Spleen
AuNS
43. Optical projection tomography
VCAM targeting in ApoE mice
A B
A diagnostic
technique to identify
fragile plaques and
guide personalised
treatment
C Control ApoE-/- D
44. Drug Targeting using Magnetic Systems
Professor Sylvain Martel
A clinical platform designed to agglomerate and
control
flagellated Magnetotactic Bacteria (type MC-1)
acting as micro-carriers
Magnetotaxis
SYSTEM
45. Self-assembled and self-reproducing
computer-controlled
biological bacterial carriers
Drug Delivery to
Colorectal Cance
Flagella acting as the
avoiding
propulsion system
Radiotherapy and
MC-1 flagellated magnetotactic bacterium Surgery
that can be directed towards the tumor
ue to a chain of nanoparticles synthesized in the cell
and acting like a miniature driven wheel
N-38 drug molecules encapsulated in liposomes
and attached to the bacterial cell
46. TMMC Drug inside TMMC
Drug Targeting in
Vessels Driven by
TMMC
MRI controlled
Magnetic Published in Biomaterials 2011
47. Nanomedicine Summary
•Nano & Biology are a Natural Fit
•Nano = chemistry, physics, engineering
requires multidisciplinary teams to address
complex issues of bio-efficacy and safety
•A relatively mature field with established
markets and products, but in rapid expansion
•Need for a balanced approach to regulate uniformly
between countries and foster development
Notas do Editor
Marrow stimulation techniques such as drilling and microfracture give rise to repair tissues that are fibrocartilaginous and non durable.