Download hinojal-zazo-gamez-universidad-of-salamanca

GOLD NANOPARTICLES FOR
TARGETING OF STAVUDINE
TO MACROPHAGE
Hinojal Zazo
Department of Pharmaceutical Sciences. Area of Pharmacy and
Pharmaceutical Technology
University of Salamanca, Spain
HIV EPIDEMIOLOGY
Zazo H.
PLoS One. 2013 Dec 18;8(12):e81355
Introduction
2
HIV PROGRESION
Zazo H.
HAART therapy can´t eliminate
all HIV viral load
PROBLEM
HAART Initiation
Plasma HIV Load
HIV-DNA Load
Trends Immunol. 2011 Mar;32(3):131-7
Introduction
3
HIV RESERVOIRS
Zazo H.
There are reservoirs with lower virus production but higher
cell longevity where drugs doesn’t reach successfully
Nature Medicine 9, 853 - 860 (2003)
Introduction
4
Zazo H.
OBJECTIVE
Drug delivery systems, such as gold
nanoparticles, have been proposed to increase
drug intracellular penetration
Stavudine gold nanoparticles (d4T-AuNP) have
been developed to target the drug into
macrophages
5
Zazo H.
METHODS
1. Preparation of stavudine gold nanoparticles.
• To check that stavudine was coupled to the AUNPs
 UV-vis spectroscopy
 Dynamic Light Scattering
 Ultra-presure chromatography with UV spectrosopy (UPLC-UV)
Materials and Methods
6
Zazo H.
METHODS
1. Preparation of stavudine gold nanoparticles.
2. Macrophages isolation and culture with:
A. Different concentrations of gold nanoparticles
free stavudine
B. Different comparable concentrations of
stavudine-AuNP
Materials and Methods
7
Zazo H.
METHODS
1. Preparation of stavudine gold nanoparticles.
2. Macrophages isolation and culture with the drug delivery system.
• Uptake
• AuNPs concentration
• Drug concentration
Transmission electron microscopy
(TEM)
Inductively coupled plasma
mass spectroscopy
(ICP-MS)
Ultra-presure chromatography UV
spectrosopy
(UPLC-UV)
Materials and Methods
8
Zazo H.
1. NPs CHARACTERIZATION
The results of gold nanoparticles characterization showed the
presence of stavudine coupled
Results and Discussion
9
Zazo H.
1. NPs CHARACTERIZATION
The results of gold nanoparticles characterization showed the
presence of stavudine coupled
Drug Loading
Zeta Potential
-64.30
-28.67
D4T Coupled
(mg d4T/mL)
Loading
efficacy (%)
0.729 (±0.04)
70.9
Results and Discussion
10
Zazo H.
2. UPTAKE STUDY
Human primary monocyte-derived macrophages were
incubated for 24 hours with gold nanoparticles
0.025 ug Au/mL
0.25 ug Au/mL
2.5 ug Au/mL
Results and Discussion
25 ug Au/mL
11
Zazo H.
TEM ANALYSIS
Results and Discussion
12
Zazo H.
2A. NAKED NANOPARTICLES UPTAKE
Nanoparticles concentration in
the medium influenced the
% AuNP into the cells
kinetic uptake
*
60
40
20
Results and Discussion
P
uN
A
0.
02
5
A
uN
P
0.
25
A
uN
P
0
2.
5
% Au/million cells
80
13
Zazo H.
2A. NAKED NANOPARTICLES UPTAKE
Gold nanoparticles
Stavudine
Results and Discussion
14
Zazo H.
2B. STAVUDINE UPTAKE
Stavudine uptake after 24h is
deeply higher using gold
nanoparticles as carriers
% stavudine into cells.
***
***
***
50
fr
ee
-d
4T
1
d4
TA
uN
10
P
fr
ee
-d
10
4T
d4
TA
uN
10
P
0
fr
ee
-d
10
4T
0
d4
TA
uN
P
0
1
% stavudine uptake
100
Results and Discussion
15
Zazo H.
CONCLUSION
Gold nanoparticles are a
promising nanotechnological
strategy for targeting drugs to
macrophages.
Conclusion
16
Zazo H.
ACKNOWLEDGEMENTS
Pharmacy and Pharmaceutical Technology Area.
Medizinische Klinik III. Prof. Tacke Lab
(Pharmaceutical Sciences. University of Salamanca)
17
THANK YOU
FOR YOUR ATTENTION
[email protected]
THANK YOU
FOR YOUR ATTENTION
[email protected]