2. Premises Since the human body is basically an extremely
complex system of interacting molecules (i.e., a molecular
machine), the technology required to truly understand and repair
the body is the molecular machine technology : NANOTECHNOLOGY A
natural consequence of this level of technology will be the ability
to analyze and repair the human body as completely and effectively
as we can repair any conventional machine today.
3. MAJOR BIOLOGICAL STRUCTURES IN SCALE
4. NANO < 100 nm
5. Nanomedicine:EC European Technology Platform (ETP)
6. E.C.-ETPNanomedicine, is defined as the application
ofnanotechnology to achieve breakthroughs inhealthcare. It exploits
the improved and often novelphysical, chemical and biological
properties ofmaterials at the nanometer scale. Nanomedicinehas the
potential to enable early detection andprevention, and to
essentially improve diagnosis,treatment and follow-up of
diseases..Diagnostics, targeted drug delivery andregenerative
medicine constitute the coredisciplines of nanomedicine.
7. Nanomedicine:European Science Foundation (ESF) The field of
Nanomedicine is the science and technology of diagnosing, treating
and preventing disease and traumatic injury, of relieving pain, and
of preserving and improving human health, using molecular tools and
molecular knowledge of the human body. It embraces sub- disciplines
which are in many ways overlapping and are underpinned by common
technical issues.
8. The numbers of nanomedicine The total market for
nanobiotechnology products is $19.3 billion in 2010 and is growing
at a compound annual growth rate (CAGR) of 9% to reach a forecasted
market size of $29.7 billion by 2015.
9. 1966
10. Topics in nanomedicine Therapy:Drug Delivery: Use
nanodevices specifically targeted to cells, to guide delivery of
drugs, proteins and genesDrug targeting : Whole body, cellular ,
subcellular deliveryDrug discovery : Novel bioactives and delivery
systems
11. Topics in nanomedicine Diagnosis: Prevention and Early
Detection of diseases: Use nanodevices to detect specific changes
in diseased cells and organism.
12. Nanoparticles (NP):Smart Nanostructures for diagnosis and
therapy
13. Why Nanoparticles1) Drugs, contrast agents,paramagnetic or
radiolabeledprobes can be vehiculated by NPs2) NPs can be
multi-functionalizedto confer differents features onthem
14. Why Nanoparticles Vehiculation: Drug-encapsulating
nanoparticles offer extensive control over delivery. Drugs are
protected inside NPs and are not degraded.
15. Targeting: nanoparticles control over delivery. Drugs are
concentrated to target. Less systemic toxicity. Less drug is
necessary
16. Multi-functionalization: Control over delivery location,
drug dosage, and drug release characteristics is possible
17. An ideal Multi-functional nanoparticle vector Anticorpo
Polietilenglicol Evita che NP venga digerita nei lisosomi Indirizza
la NP verso un la NP venga Evita che antigene specifico sulla dal
circolo rimossa cellula da colpire Tat peptide Determina Fusione e
Probe magnetico ingresso della NP nella cellula Permette imaging
tramite MRI
18. Examples of nanoparticulate carriers
19. Carbon-based: Buckyballs and Nanotubes C60 1nm
20. Carbon nanotubes can be thought of as a sheet of
graphiterolled into a cylinderNanotubes have a very broad range of
electronic, thermal, andstructural properties that change depending
on diameter,length,). They exhibit extraordinary strength and
uniqueelectrical properties, and are efficient conductors of
heat.Used as sensors
21. Proposed as a vessel for transporting drugs into the
body.The drug can be attached to the side or trailed behind, or the
drug can actually be placed inside the nanotube Nanotube
Nanocap
22. Funzionalizzazione covalenteFunzionalizzazione non
covalente
23. Funzionalizzazione non covalente (DNA)
24. Their final usage, however, may be limited by
theirpotential toxicity.Under some conditions, nanotubes can cross
membranebarriers and can induce harmful effects:
inflammation,epithelioid granulomas (microscopic
nodules),fibrosis,and biochemical/toxicological changes in
thelungs.
27. gold nanoparticles (1-20 nm) are produced by reductionof
chloroauric acid (H[AuCl4]), To the rapidly-stirred boiling HAuCl4
solution, quickly add 2 mL of a 1% solution of trisodium citrate
dihydrate, Na3C6H5O7.2H2O. The gold sol gradually forms as the
citrate reduces the gold(III). Remove from heat when the solution
has turned deep red or 10 minutes has elapsed.
28. filmato
29. In cancer research, colloidal gold can be used to
targettumors and provide detection using SERS (SurfaceEnhanced
Raman Spectroscopy) in vivo.They are being investigated as
photothermal convertersof near infrared light for in-vivo
applications, as ablationcomponents for cancer, and other targets
since nearinfrared light transmits readily through human skin
andtissue
30. Polymeric/Dendrimers(e.g.PLGA, PAA, PACA) spherical
polymers of uniform molecular weight made from branched monomers
are proving particularly adapt at providing multifunctional
modularity.
33. Dendrimers are repetitively branched molecules. PAA= POLI
AMMINO AMMIDEPLGA PAA
34. HYDROGELSPolymers or co-polymers (e.g. acrylamide and
acrylic acid) createwater-impregnated nanoparticles with pores of
well-defined size.Water flows freely into these particles, carrying
proteins and other smallmolecules into the polymer matrix.By
controlling the pore size, huge proteins such as albumin
andimmunoglobulin are excluded while smaller peptides and
othermolecules are allowed.The polymeric component acts as a
negatively charged "bait" that attracts positivelycharged proteins,
improving the particlesperformance.
35. Mesoporoussilica (SiO2)
36. Mesoporous silica particles: nano-sized spheres or rods
filled with a regulararrangement of pores with controllable pore
size from 3 to 15nm and outer diameterfrom 20nm to 1000 nm .The
large surface area of the pores allows the particles to be filled
with a drug or with a fluorescent dye that would normally be unable
to pass through cell walls.The MSN material is then capped off with
a molecule that is compatible with thetarget cells. When are added
to a cell culture, they carry the drug across the cellmembrane.
These particles are optically transparent, so a dye can be seen
throughthe silica walls. The dye in the particles does not have the
same problem with self-quenching that a dye in solution has. The
types of molecules that are grafted to theoutside will control what
kinds of biomolecules are allowed inside the particles tointeract
with the dye. EM
38. A quantum dot is a semiconductor whose excitons areconfined
in all three spatial dimensions.An immediate optical feature of
colloidal quantumdots is their colorationFirst attempts have been
made to use quantum dotsfor tumor targeting under in vivo
conditions.Generically toxic
39. Quantum Dots Raw quantum dots, 2-8 nm are toxic, CdSe or
CdTe cores with ZnS shell But they fluoresce brilliantly, better
than dyes (imaging agents) Only way of clearance of protected QDs
from the body is by slow filtration and excretion through the
kidneyhttp://www.azonano.com/Details.asp?ArticleID=1726
40. Quantum DotsQD technology helps cancer researchers to
observe fundamentalmolecular events occurring in the tumor cells by
tracking theQDs of different sizes and thus different colors,
tagged tomultiple different biomoleules, in vivo by
fluorescentmicroscopy.QD technology holds a great potential for
applications innanobiotechnology and medical diagnostics where QDs
could beused as labels.
41. Nano-particulate pharmaceuticalsBrand name DescriptionEmend
Nanocrystal (antiemetic) in a capsule(Merck & Co. Inc.)Rapamune
Nanocrystallized Rapamycin (immunosuppressant) in a(Wyeth-Ayerst
Laboratories) tabletAbraxane Paclitaxel (anticancer drug)- bound
albumin particles(American Biosciences, Inc.)Rexin-G A retroviral
vector carrying cytotoxic gene(Epeius Biotechnologycorporation)Olay
Moisturizers Contains added transparent, better protecting nano
zinc(Procter and Gamble) oxide particlesTrimetaspheres (Luna
Nanoworks) MRI imagesSilcryst Enhance the solubility and sustained
release of silver(Nucryst Pharmaceuticals) nanocrystalsNano-balls
Nano-sized plastic spheres with drugs (active against(Univ. of
South Florida) methicillin-resistant staph (MRSA) bacteria)
chemically bonded to their surface that allow the drug to be
dissolved in water.
42. Company ProductCytImmune Gold nanoparticles for targeted
delivery of drugs to tumorsNucryst Antimicrobial wound dressings
using silver nanocrystalsNanobiotix Nanoparticles that target tumor
cells, when irradiated by xrays thenanoparticles generate electrons
which cause localized destruction of the tumor cells.Oxonica
Disease identification using gold nanoparticles
(biomarkers)Nanotherapeutics Nanoparticles for improving the
performance of drug delivery byoral, inhaled or nasal
methodsNanoBio Nanoemulsions for nasal delivery to fight viruses
(such as the flu andcolds) and bacteriaBioDelivery Sciences Oral
drug delivery of drugs encapuslated in a nanocrystallinestructure
called a cochleateNanoBioMagnetics Magnetically responsive
nanoparticles for targeted drugdelivery and other
applicationsZ-Medica Medical gauze containing aluminosilicate
nanoparticles which help boodclot faster in open wounds.
43. Some liposome -based pharmaceuticals
44. Open ProblemsManufacturing NPs for medical SOLUTION: use:
Assessment of NPs: Putting the drug on the particle Dynamic
structural Maintaining the drug on the features in vivo particle
Kinetics of drug Making the drug come off the release particle once
application is Triggered drug release done Purity and homogeneity
of nanoparticles
45. Open ProblemsToxicity:short term - no toxicity in
animalslong term- not knownToxicity for both the host and the
environment should be addressed
46. Open ProblemsDelivery: SOLUTION: Ensuring Delivery to
target detection of NPs organ/cell at target, organs , Removal of
nanoparticles from cells , subcellular the body location et al.
Tissue distribution
47. Open Problems: Targeting the brain Brain micro-vessel
endothelial cells build up the blood brain barrier (BBB) The BBB
hinders water soluble molecules and those with MW > 500 from
getting into the brain
48. The blood-brain barrier (BBB)NPs
49. Open Problems GMP Challenges No standards for: Purity and
homogeneity of nanoparticles Manufacturing Methods Testing and
Validation
50. Summary Toxicities of nanomaterials are unknown How to best
target the nanomaterials so that systemic administration can be
used ? How to uncage the drug so it gets out at the desired
location ? Is there a way to re-cage the drug when it is no longer
desired ? How are nanoparticles removed from the body ?
Mathematical modeling of nanostructures is in its infancy Barrier
crossing (BBB, G.I., et al.)
51. Nanomedicine programs atDepartment of Experimental Medicine
(DIMS) Universit di Milano-Bicocca
52. Nanoparticles for therapy and diagnosis of Alzheimer
Disease(NAD)- European Community September 2008-August 2013
Synthesis of new NPs with improved biocompatibility, targeting and
drug delivery features - Cariplo Bank Foundation Nanoparticles
safety
53. A Nanochip to test the activity of candidate anti-
amyloidogenic drugsOverlay (reflection and fluorescence)
confocalimages of labelled Abeta aggregates innanochannels.1
cross-section 260 nm 300 nm. 2 cross section 310 nm 300.Sordan R.
et al, Vertical arrays of nanofluidic channels fabricated
withoutnanolithography Lab on a Chip,2009
54. NAD: NANOPARTICLES FOR THERAPY AND DIAGNOSIS OF ALZHEIMER
DISEASE 2008-2012 Funding scheme: COLLABORATIVE PROJECT Large-scale
integrating project (IP) proposal Work programme topic addressed:
FP7-NMP-2007-LARGE-1NMP-2007-4.0-4 Substantial innovation in the
European medical industry: development of nanotechnology-based
systems for in-vivo diagnosis and therapy
55. No Partner Country Role Person1 University of
Milano-Bicocca, Milano Italy Ligand generation, Binding Massimo
Masserini BiocompatibilityCharacterization2 Universidad Autonoma de
Madrid, Madrid Spain Transgenic models of AD Francisco Wandosel3
University of Brighton, Brighton UK Biomimetics Moghimi S Moein4
University of Paris-Sud XI, Chatenay- France Polymeric NPs Karine
Andrieux Malabry5 Slovak Academy of Sciences, Bratislava Slovakia
Computational models Igor Tvaroska6 Karolinska Institutet,
Stockholm Sweden Neurobiology of AD Jin Jing Pei7 Academic Medical
Center, Amsterdam The Neurobiology of AD Wiep Scheper Netherlands8
Biotalentum Ltd , Gdll Hungary Management Andras Dinnyes9 Turku PET
Center, Turku Finland PET Juha Rinne10 Nanovector Srl, Torino Italy
SLN Gasco Mariarosa11 University Of Patras, Patras Greece Liposomes
Sophia G. Antimisiaris12 University of Antwerp, Antwerp Belgium MRI
Annemie Van der Linden13 Universidad del Pas Vasco, Bilbao Spain
Biophysics Felix Goni14 Lancaster University, Lancaster UK
Neurobiology David Allsop15 I.R.F. "Mario Negri, Milan Italy
Pharmacokinetics Mario Salmona16 Stab Vida, Oieras Portugal
Antibodies Bruno Mateus17 Universit Pierre et Marie Curie, Paris
France Post-mortem samples Charles Duyckaerts18 GUERBET SA,
Villepinte France Imaging Marc Port19 Bial Industrial Farmacutica
S.A., Bilbao Spain Immunogenicity Alberto Martnez
56. ALZHEIMER DISEASE NEL : 24 milioni di casi di demenza 4.6
milioni nuovi casi/anno (1 ogni 7 secondi). EU: Circa 5 milioni di
casi di demenza Pi di 3 milioni di Alzheimer (AD). 2x entro il 2040
in Europa occidentale 3x nell Europa dellest
57. La produzione progressiva e laccumulo di b-amiloide (Ab),
un frammento della Amyloid Precursor Protein (APP), svolgono un
ruolo centrale nella malattia di Alzheimer. APP
58. Ab aggrega formandoOligomeri, fibrille, placche
59. Ab induce neurodegenerazione plaque
60. IL PROGETTO NAD Legare Abeta !
61. IL PROGETTO NAD 1- DISGREGAZIONE e IMAGING di placche di Ab
nel cervello: terapia e diagnostica combinate (theranostics) in
modelli animali di AD.2- CLEARANCE of Ab dal sangue, richiamando
leccessoanche dal cervello (sink effect) in modelli animali di AD.
NANOPARTICELLE
64. FASI 1: Sintesi di NPs2: Selezione e Sintesi di ligandi per
Ab /funzionalizzazione di NP 3: Funzionalizzazione di NP per
attraversare la barriera emato-encefalica (BBB) 4:
Funzionalizzazione di NP per diagnostica (MRI, PET) 5:
Biocompatibilit 6: Prove in vitro della efficienza del sistema
(cell cultures) 7: Prove in vivo della efficienza del sistema (TG
mice)
65. 1- DISGREGAZIONE e IMAGING di placche di AbLe
nanoparticelle saranno funzionalizzate per legare Ab e per MRI or
PET imaging Ab NPs aggregates
66. NANOPARTICELLE E BARRIERA EMATOENCEFALICA Ab AGGREGATES
Capillari cerebrali blood FLUSSO DEL SANGUE Le Nanoparticelle
saranno funzionalizzate per superare la barriera emato encefalica
(BBB).Entrando nel cervello per endo/transcitosi
67. 2- RIMOZIONE DI Ab DAL SANGUE ( SINK EFFECT )BRAIN
b-AMYLOID AGGREGATES EQUILIBRIUM BRAIN/BLOOD SHIFT Blood-brain
barrier AbNANOPARTICLES BLOOD CIRCULATION RES + LIVER
68. NAD OBJECTIVES1- DISGREGATION and IMAGING of brain Ab :
combined therapy e diagnostics (theranostics) in animal models of
AD.