1. Nanoparticle Delivery
N i l D li
 Oral
O l
 Intravenous
 Nasal
 Ophthalmic
 Rectal/Vaginal
 Trans Dermal
 Pulmonary

2. Local delivery vs. systemic
 Aerosol route of administration
 Local effect (reduced systemic site effects)
( y )
 Systemic (prevent Gi-tract or invasive administration)
 Aerosol target
 Treatment Organ
 Absorption Organ
p g

3. Lungs as Treatment organ
 Asthma (Hardy and Chadwick, 2000)
 Cystic fibrosis (Garcia-Contreras and
Hickey, 2002)
 Lung cancer (Rao et al 2003)
al.,
 Tuberculosis (Pandey and Khuller,
2005; Zahoor et al., 2005)
h l )

4. NANOPARTICLES in Cancer
Treatment
 Nanoparticles are drug vehicles able to
accumulate in tumor tissues or cells, while
protecting the drug from premature
inactivation during the transport…
g p
 Promising drug carriers…
Brigger I, Dubernet C, Couvreur P. Nanoparticles in cancer therapy
and diagnosis. Advanced Drug Delivery Reviews 2002;54(5):631-651.

5. Biological activity of the
g y
Nanoparticles
H460 A549
100 100
Dox-NP
Dox-NP Empty-NP
py
80
xicity （ % ）
80 Empty-NP
E t NP Dox
otoxicity (%)
Dox
60 60
40 40
Cytotox
Cyto
20 20
0
0
0.625 0.315 0.156 0.078 0.039 0.019
0 625 0 315 0 156 0 078 0 039 0 019
0.625 0.315 0.156 0.078 0.039 0.019
0 625 0 315 0 156 0 078 0 039 0 019
Doxorubicin ( g/mL)
Doxorubicin ( g/mL)
Shirzad Azarmi, Xia Tao, Hua Chen, Zhaolin Wang, Warren. H. Finlay,
Raimar Löb b
R i Löbenberg, Wil
Wilson. H. R
H Roa; F
Formulation and C t t i it of D
l ti d Cytotoxicity f Doxorubicin Nanoparticles
bi i N ti l
Carried by Dry Powder Aerosol Particles
Int J Pharm 2006

6. DEPOSITION MECHANISM
General air flow through the
respiratory tract
Large particles will Impact
in the upper respiratory
tract (>5 um)
Small particles will have
no deposition because of
Brownian diffusion
( um)
(<1 u )
Sedimentation in the
alveolar region (1-5 um)
(1 5
www.becomehealthynow.com

7. Pulmonary Nanoparticle delivery
 Solid colloidal particles < 1000 nm
 But how can you deliver them to the lungs?
 Suitable size is between 1000 and 5000 nm?
We had to go back to the lab
and do some experiments

8. THE CONCEPT
Respirable aerosol
particles
containing
nanoparticles
The carrier particles
deposit in the alveolar region
The carrier dissolves
The nanoparticles are released

9. THE CONCEPT
evaporation of water Respirable
lactose aerosol
Suspension of particles
lactose and containing
nanoparticles nanoparticles
Spray drying

10. RESULTS
A B C
CARRIER PARTICLES NANOPARTICLES CARRIER PARTICLES
and NANOPARTICLES

11. CUTTING THROUGH THE
PARTICLES
Continuous matrix of carrier particles
Confocal laser
scanning
microscopy
Hollow carrier particles

12. Most Cited Paper

13. Active Release of Nanoparticles
from the Carrier Particle
 How to create forces which increase the
disperse-ability of the nanoparticles?
We had to go back to the lab
and do some experiments

14. NEW SYSTEM OF CARRIER
PARTICLES with Active Release
Our approach:
 The in situ generated gas
pressure may h l to
help t
disperse the content of
the carrier particle

15. Active Release of Nanoparticles
from the Carrier Particle
 How to how to avoid that the reaction is
happening while spray drying?
We had to go back to the lab
and do some experiments

16. NEW SYSTEM OF CARRIER
PARTICLES
 Effervescent carrier particles:
Lactose
Sodium Carbonate
FORMULATION Acid Citric
Ammonia Evaporation
Water

17. MAJOR PARAMETERS
Machine parameters
Pump
Atomization
•Pressure
Temperatures
•inlet Aspirator
•outlet

18. MAJOR PARAMETERS
Formulation parameters
Solid concentration
Excipients:
E i i t
• leucine
• polyethylene glycol
• ethanol
• surfactants

19. RESULTS: Polyethylene glycol
(PEG) and L-leucine
PEG and L-leucine
50
w e ight fra ction %
40
30
20
10
0
5.6 4.3 3.4 2 1.1 0.51
cut points
PEG and l-leucine both work as a lubricant
L- leucine can decrease the aggregation of
gg g
spray dried particles

20. Fine Particle Fraction (FPF)
Fine Particle Fraction
50 43.97
45
40 35.7
35
FPF (%
30
F
25
17.6
20
15 12.5 12.5
10
5
0
Tween 80 Ethanol 30% Lactose 1% Spray dryer L-leucine +
and Jet mill PEG
FPF means particle fraction that can be delivered to the lungs

21. Carrier particles with
active release mechanism
A B
(A) green nanoparticles
nanoparticles.
(B) red carrier matrix
(C) normal light
(D)
C D

22. Carrier particles with
active release mechanism
After the
sample wasas The green
exposed to Nanoparticles
humidity are actively
distributed in
the gas bubble
The red carrier
matrix
dissolves
The gas bubble is about 30 m

23. RESULTS: Influence of the
formulation on the size of nanoparticles
350
Effervescent Dry Powder 300
for Respiratory Drug
Delivery; 250
*
Leticia Ely, Wilson Roa,
Warren H. Finlay, Raimar 200
nm
Löbenberg;
Eur Pharm Bi h
E J Ph Biopharm 150
2007
100
50
0
re
r
re
te
r
fo
te
fo
Af
Be
Af
Be
nt
e
nt
ce
os
e
ce
os
es
ct
es
ct
La
rv
rv
La
fe
fe
Ef
Ef

24. Publication

25. Nanotoxicology
of Inhalable Nanoparticles
IS it safe to use inhalable
nanoparticles?

26. Lung surfactant
 Lung surfactant are a mixture of about 80%
phospholipids,
 5-10 proteins and
 5-10% cholesterol containing compounds
g p
 Major co po e ts o t e p osp o p ds a e
ajo components of the phospholipids are
phosphatidylcholine PC and dipalmitoyl
phosphatidylcholine DPPC.

27. Lung surfactant
Function:
 Reduction of Surface Tension
 Gas Exchange

28. Surface pressure versus
molecular area isotherms
dipalmitoyl phosphatidylcholine DPPC
DPPC nanoparticles
313 nm
187 nm

29. Surface pressure versus
molecular area isotherms
The study concluded
3 that:
th t The high surface
2 pressure values obtained from
1 187 nm NP the isotherms of the binary
1 2 230 nm NP mixtures indicates the notion
3 DPPC that their size dependent
incorporation does not
destabilize the monolayer
film. The study results
demonstrated that pulmonary
nanoparticle delivery is a
possible route of
administration.
Biophysical investigation of nanoparticle interactions with lung surfactant model
systems; Diana Stuart, Raimar Löbenberg, Tabitha Ku, Shirzad Azarmi, Leticia Ely,
Wilson Roa and Elmar J. Prenner J Biomed Nanotech Vol 2 p1-8 2006

30. Spray freeze-drying
Nitrogen Gas
Peristaltic
Doxorubicin pump Particles deposit in the
nanoparticles alveolar region. The
lactose
carrier dissolves and
nanoparticles are
ti l
released.
Carrier Powder
Liquid
Nitrogen Freeze Drying
72 hr
300-400 mL 1-5 µm
µ
Density: ~ 0.1 g/cm3

31. Project/Animal Identification: Score
Morbidity
Appearance
Normal 0
General lack of grooming
g g 1
Scores
Coat staring, ocular or nasal discharge 2
Piloerection, hunched up 3
Body Weight
Normal < 5% 0
body wt. drop 6-15% 1
body wt. drop 16-25% 2
25 Body weight 14 body wt. drop 26-35% 3
body wt. drop > 35%
y p 4
Morbidity
M bidit score 12 Food Intake
20
Normal 0
10
B o d y w e ig h t (g )
M o rb i d i ty s c o r e
food intake drop 10-33% 1
15 8 food intake drop 34-75%
p 2
food intake drop > 75% 3
10 6 Clinical Signs
Normal resp. rate and hydration 0
4 Slight changes
g g 1
5 Resp. rates up or down 30%, measurable dehydration 2
2
Resp. rates changes 50% or very low, severe dehydration 3
0 0 Behavior
1 8 15 22 29 36 43 50 Normal 0
Minor inactivity or exaggerated responses 1
Time (day)
Moderate change in expected behavior, isolated or listless 2
Reacts violently, or very weak and precomatose 3
Total

32. Study design
 Female 4-5 weeks old BALB/c nude mice
 NCl-H460 injection
 Small lung metastatic nodules develop in 15
days
 Treatment over 4 weeks with
T k ih
 Free Drug
 Inhalable NPs
 Blank NPs
 No treatment

33. Expectation
 The cancer will spread throughout the body
 The animals will die because of cancer in
other organs
 The Lungs might show less cancer
compared to other tissues due to the
p
treatment

34. Summary of what accomplished
and performed
Assessing the efficacy of DOX-loaded effervescent
inhalable NPs in vivo
100%
90%
80%
70%
rvival %
60%
50%
Surv
40%
30% Effervescent NPs
Normal NPs
20% DOX-NP IV
10% Dox solution IV
Control
0%
0 20 40 60 80 100 120 140

35. Summary
 Inhalable nanoparticles with active release mechanism
were successfully synthesized
 In vitro results demonstrate that nanoparticles do not
compromise the biological function of the lung surfactant
film
 In vitro cell culture test show that doxorubicin bound to
nanoparticles is more effective against lung cancer cells
than free doxorubicin
 In vivo studies demonstrate that the survival of mice can
be improved if doxorubicin loaded nanoparticles are
administered via the pulmonary route of administration

36. Conclusions
 New dry powder technology is available to
deliver NP or drugs to the lungs
 Improved pulmonary drug delivery for
cytotoxic molecules is possible using
nanoparticles
 Drug d li
D delivery i th k t i
is the key to improve (l
(lung
cancer) treatments.

37. Articles
 Formulation and in vivo evaluation of effervescent inhalable carrier particles for pulmonary delivery of
nanoparticles.
Azarmi S. Lobenberg R. Roa WH. Tai S. Finlay WH.
Drug Development & Industrial Pharmacy. 34(9):943-7, 2008 Sep.
 Targeted delivery of nanoparticles for the treatment of lung diseases
Shirzad Azarmi, Wilson H. Roa and Raimar Löbenberg,
Advanced Drug Delivery Reviews, 0(8):863-75, 2008 May 22.
 Nanoparticles: characteristics, Mechanisms of Action and Toxicity in Pulmonary Drug Delivery – A Review
S Gill R Löbenberg T Ku S Azarmi W Roa and EJ Penner
Gill, Löbenberg, Azarmi,
J Biomed Nanotechnology 2007
 Effervescent Dry Powder Aerosols for Respiratory Drug Delivery
Leticia Ely, Warren H Finlay, Wilson H Roa, Raimar Löbenberg
Eur J Pharm Biopharm 2007 (65) 346-353
 Biophysical investigation of nanoparticle interactions with lung surfactant model systems
Diana Stuart, Raimar Löbenberg, Tabitha Ku, Shirzad Azarmi, Leticia Ely, Wilson Roa and Elmar J. Prenner
J Biomed Nanotechnology Volume 2, Numbers 3-4, October/December 2006, pp. 245-252(8)
 Formulation and Cytotoxicity of Doxorubicin Nanoparticles Carried by Dry Powder Aerosol Particles
Shirzad Azarmi, Xia Tao, Hua Chen, Zhaolin Wang, Warren. H. Finlay, Raimar Löbenberg, Wilson. H. Roa
Int J Pharm 2006 319 (1-2), pp. 155-161 (ranked # 23 for the journal in Jul – Sep 2006 )
 Optimization of a two step desolvation method for preparing gelatin nanoparticles and cell uptake studies in 143B
two-step
osteosarcoma cancer cells
Azarmi S, Huang Y, Chen H, McQuarrie S, Abrams D, Roa W, Finlay WH, Miller GG, Löbenberg R
JPPS 9 (1): 124-132 2006
 Dry Powder Inhalation Aerosols Containing Nanoparticulate Doxorubicin
L.G. Sweeney, H. Chen, Z. Wang, R. Löbenberg, W. Roa, W.H. Finlay
y g g y
Eur J Pharm Sci 305 (1-2): 180-185 NOV 23 2005
 Formulation and Characterization of Spray-Dried Powders Containing Nanoparticles for Aerosol Delivery to the Lung,
Jeffrey O.-H. Sham, Yu Zhang, Warren H. Finlay, Wilson H. Roa, and Raimar Löbenberg
Int. J. Pharm. Vol 269, (2), Pages 457-467 ranked Top 25 in 2004

38. Acknowledgements
 Dr. Finlay
D Fi l
 Dr. Roa
 Dr. Chacra
 Dr. Prenner
 Dr. Shirzad Azarmi
 Dr.
Dr Xia Tao
 Dr. Chuching Tai
 Yuan Huang
 Leticia Ely
L ti i El
 Kamal Al-Hallak
 NSERC, AHFMR, CIHR, ACB