This document discusses nanomedicine and its potential applications for diagnosis and treatment of diseases like Alzheimer's disease. It begins by explaining how nanotechnology allows analysis and repair of the body at the molecular level similarly to how machines are repaired today. It then discusses various nanoscale structures and materials that can be used for nanomedicine, such as liposomes, dendrimers, mesoporous silica, quantum dots, carbon nanotubes, and polymers. Examples are given of current nanomedicine products and applications being researched include drug delivery, imaging, and regenerative medicine. However, challenges are also noted around manufacturing nanoparticles for medical use, assessing their toxicity, ensuring targeted delivery, and removing nanoparticles from the body

1. Nanomedicine

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.-ETP
“Nanomedicine, is defined as the application of
nanotechnology to achieve breakthroughs in
healthcare. It exploits the improved and often novel
physical, chemical and biological properties of
materials at the nanometer scale. Nanomedicine
has the potential to enable early detection and
prevention, and to essentially improve diagnosis,
treatment and follow-up of diseases.
……………………….
Diagnostics, targeted drug delivery and
regenerative medicine constitute the core
disciplines 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.

10. 1966

11. Topics in nanomedicine
• Therapy:
Drug Delivery: Use nanodevices specifically
targeted to cells, to guide delivery of drugs,
proteins and genes
Drug targeting : Whole body, cellular ,
subcellular delivery
Drug discovery : Novel bioactives and
delivery systems

12. Topics in nanomedicine
• Diagnosis:
Prevention and Early Detection of diseases: Use
nanodevices to detect specific changes in diseased
cells and organism.

13. Nanoparticles (NP):
Smart Nanostructures for diagnosis
and therapy

14. Why Nanoparticles
1) Drugs, contrast agents,
paramagnetic or radiolabeled
probes can be vehiculated by NPs
2) NPs can be multi-functionalized
to confer differents features on
them

15. Why Nanoparticles
• Vehiculation: Drug-encapsulating
nanoparticles offer extensive control over
delivery.
• Drugs are protected inside NPs and are not
degraded.

16. • Targeting: nanoparticles control over
delivery.
• Drugs are concentrated to target. Less
systemic toxicity.
• Less drug is necessary

17. • Multi-functionalization: Control over delivery
location, drug dosage, and drug release
characteristics is possible

18. 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

19. • Examples of nanoparticulate
carriers

20. Carbon-based: Buckyballs and
Nanotubes
C60 1nm

21. Carbon nanotubes can be thought of as a sheet of graphite
rolled into a cylinder
Nanotubes have a very broad range of electronic, thermal, and
structural properties that change depending on diameter,
length,). They exhibit extraordinary strength and unique
electrical properties, and are efficient conductors of heat.
Used as sensors

22. 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

23. Funzionalizzazione covalente
Funzionalizzazione non covalente

24. Funzionalizzazione non covalente (DNA)

25. Their final usage, however, may be limited by their
potential toxicity.
Under some conditions, nanotubes can cross membrane
barriers and can induce harmful effects: inflammation,
epithelioid granulomas (microscopic nodules),
fibrosis,and biochemical/toxicological changes in the
lungs.

26. Lipid-based NPs :Liposomes and
solid lipid nanoparticles (SLN)
50 – 500 nm 40-1000nm

27. Metal-core nanoparticles

28. gold nanoparticles (1-20 nm) are produced by reduction
of 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.

29. filmato

30. In cancer research, colloidal gold can be used to target
tumors and provide detection using SERS (Surface
Enhanced Raman Spectroscopy) in vivo.
They are being investigated as photothermal converters
of near infrared light for in-vivo applications, as ablation
components for cancer, and other targets since near
infrared light transmits readily through human skin and
tissue

31. 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.

32. Polymeric
PLGA ACIDO POLI-LATTICO-GLICOLICO
PLGA Poly-Lactic-Glycolic Acid

33. In solvente organico In acqua

34. Dendrimers are repetitively
branched molecules.
PAA= POLI
AMMINO AMMIDE
PLGA
PAA

35. HYDROGELS
Polymers or co-polymers (e.g. acrylamide and acrylic acid) create
water-impregnated nanoparticles with pores of well-defined size.
Water flows freely into these particles, carrying proteins and other small
molecules into the polymer matrix.
By controlling the pore size, huge proteins such as albumin and
immunoglobulin are excluded while smaller peptides and other
molecules are allowed.
The polymeric component acts as a negatively
charged "bait" that attracts positively
charged proteins, improving the particles'
performance.

36. Mesoporous
silica (SiO2)

37. Mesoporous silica particles: nano-sized spheres or rods filled with a regular
arrangement of pores with controllable pore size from 3 to 15nm and outer diameter
from 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 the
target cells. When are added to a cell culture, they carry the drug across the cell
membrane. These particles are optically transparent, so a dye can be seen through
the 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 the
outside will control what kinds of biomolecules are allowed inside the particles to
interact with the dye.
EM

38. Fluorescent and paramagnetic
(e.g.Quantum dots)
• Crystalline fluorophores
• CdSe semiconductor core
• ZnS Shell
3 nm

39. A quantum dot is a semiconductor whose excitons are
confined in all three spatial dimensions.
An immediate optical feature of colloidal quantum
dots is their coloration
First attempts have been made to use quantum dots
for tumor targeting under in vivo conditions.
Generically toxic

40. 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 kidney
http://www.azonano.com/Details.asp?ArticleID=1726

42. Quantum Dots
QD technology helps cancer researchers to observe fundamental
molecular events occurring in the tumor cells by tracking the
QDs of different sizes and thus different colors, tagged to
multiple different biomoleules, in vivo by fluorescent
microscopy.
QD technology holds a great potential for applications in
nanobiotechnology and medical diagnostics where QDs could be
used as labels.

44. Nano-particulate pharmaceuticals
Brand name Description
Emend Nanocrystal (antiemetic) in a capsule
(Merck & Co. Inc.)
Rapamune Nanocrystallized Rapamycin (immunosuppressant) in a
(Wyeth-Ayerst Laboratories) tablet
Abraxane Paclitaxel (anticancer drug)- bound albumin particles
(American Biosciences, Inc.)
Rexin-G A retroviral vector carrying cytotoxic gene
(Epeius Biotechnology
corporation)
Olay Moisturizers Contains added transparent, better protecting nano zinc
(Procter and Gamble) oxide particles
Trimetaspheres (Luna Nanoworks) MRI images
Silcryst Enhance the solubility and sustained release of silver
(Nucryst Pharmaceuticals) nanocrystals
Nano-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.

45. Company Product
CytImmune Gold nanoparticles for targeted delivery of drugs to tumors
Nucryst Antimicrobial wound dressings using silver nanocrystals
Nanobiotix Nanoparticles that target tumor cells, when irradiated by xrays the
nanoparticles 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 by
oral, inhaled or nasal methods
NanoBio Nanoemulsions for nasal delivery to fight viruses (such as the flu and
colds) and bacteria
BioDelivery Sciences Oral drug delivery of drugs encapuslated in a nanocrystalline
structure called a cochleate
NanoBioMagnetics Magnetically responsive nanoparticles for targeted drug
delivery and other applications
Z-Medica Medical gauze containing aluminosilicate nanoparticles which help bood
clot faster in open wounds.

46. Some liposome -based pharmaceuticals

47. Open Problems
Manufacturing 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

48. Open Problems
Toxicity:
short term - no toxicity in animals
long term- not known
Toxicity for both the host and the environment should be addressed

49. Open Problems
Delivery: 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

50. 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

51. The blood-brain
barrier (BBB)
NPs

52. Open Problems
• GMP Challenges
• No standards for:
• Purity and homogeneity of nanoparticles
• Manufacturing Methods
• Testing and Validation

53. 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.)

54. Nanomedicine programs at
Department of Experimental Medicine (DIMS)
Università di Milano-Bicocca

55. 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

56. A Nanochip to test
the activity of
candidate anti-
amyloidogenic drugs
Overlay (reflection and fluorescence) confocal
images of labelled Abeta aggregates in
nanochannels.
1 cross-section 260 nm × 300 nm.
2 cross section 310 nm × 300.
Sordan R. et al, Vertical arrays of nanofluidic channels fabricated without
nanolithography Lab on a Chip,2009

58. 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-1
NMP-2007-4.0-4 Substantial innovation in the European medical industry: development of
nanotechnology-based systems for in-vivo diagnosis and therapy

59. No Partner Country Role Person
1 University of Milano-Bicocca, Milano Italy Ligand generation, Binding Massimo Masserini
BiocompatibilityCharacterization
2 Universidad Autonoma de Madrid, Madrid Spain Transgenic models of AD Francisco Wandosel
3 University of Brighton, Brighton UK Biomimetics Moghimi S Moein
4 University of Paris-Sud XI, Chatenay- France Polymeric NPs Karine Andrieux
Malabry
5 Slovak Academy of Sciences, Bratislava Slovakia Computational models Igor Tvaroska
6 Karolinska Institutet, Stockholm Sweden Neurobiology of AD Jin Jing Pei
7 Academic Medical Center, Amsterdam The Neurobiology of AD Wiep Scheper
Netherlands
8 Biotalentum Ltd , Gödöllő Hungary Management Andras Dinnyes
9 Turku PET Center, Turku Finland PET Juha Rinne
10 Nanovector Srl, Torino Italy SLN Gasco Mariarosa
11 University Of Patras, Patras Greece Liposomes Sophia G.
Antimisiaris
12 University of Antwerp, Antwerp Belgium MRI Annemie Van der
Linden
13 Universidad del País Vasco, Bilbao Spain Biophysics Felix Goni
14 Lancaster University, Lancaster UK Neurobiology David Allsop
15 I.R.F. "Mario Negri”, Milan Italy Pharmacokinetics Mario Salmona
16 Stab Vida, Oieras Portugal Antibodies Bruno Mateus
17 Université Pierre et Marie Curie, Paris France Post-mortem samples Charles Duyckaerts
18 GUERBET SA, Villepinte France Imaging Marc Port
19 Bial Industrial Farmacéutica S.A., Bilbao Spain Immunogenicity Alberto Martínez

60. 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
Più di 3 milioni di Alzheimer (AD).
2x entro il 2040 in Europa occidentale
3x nell’ Europa dell’est

61. La produzione progressiva e l’accumulo di b-amiloide (Ab),
un frammento della Amyloid Precursor Protein (APP),
svolgono un ruolo centrale nella malattia di Alzheimer.
APP

62. Ab aggrega formando
Oligomeri, fibrille, placche

63. Ab induce neurodegenerazione
plaque

64. IL PROGETTO “NAD”
Legare Abeta
!

65. 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 l’eccesso
anche dal cervello (“sink” effect) in modelli animali di AD.
NANOPARTICELLE

66. NANOPARTICELLE(NPs)
Vantaggi:
biocompatibilità
non-immunogenicità
non-tossicità
biodegradabilità
preparazione semplice
stabilità
FUNZIONALIZZAZIONE MULTIPLA

67. NANOPARTICELLE (NPs)
LIP SLN PNP
Liposomi Solido-lipidiche Polimeriche

68. FASI
1: Sintesi di NPs
2: 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: Biocompatibilità
6: Prove in vitro della efficienza del sistema (cell cultures)
7: Prove in vivo della efficienza del sistema (TG mice)

69. 1- DISGREGAZIONE e IMAGING di placche di Ab
Le nanoparticelle saranno funzionalizzate per legare Ab
e per MRI or PET imaging
Ab
NPs aggregates

70. 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

71. 2- RIMOZIONE DI Ab DAL SANGUE ( “SINK EFFECT” )
BRAIN
b-AMYLOID
AGGREGATES
EQUILIBRIUM
BRAIN/BLOOD SHIFT
Blood-brain barrier
Ab
NANOPARTICLES BLOOD CIRCULATION RES + LIVER

72. “NAD” OBJECTIVES
1- DISGREGATION and IMAGING of brain Ab
: combined therapy e diagnostics
(theranostics) in animal models of AD.

73. 2009