Alf Lamprecht Laboratory of Pharmaceutical Engineering (EA3924), University of Franche-Comté,...
-
Upload
kerry-stokes -
Category
Documents
-
view
228 -
download
1
Transcript of Alf Lamprecht Laboratory of Pharmaceutical Engineering (EA3924), University of Franche-Comté,...
Alf Lamprecht
Laboratory of Pharmaceutical Engineering (EA3924),University of Franche-Comté, Besançon, France
Contact: [email protected]
Nanoparticulate drug delivery systems
for different therapeutic applications
“Le Menu”
A. Nanoparticles for the absorption of drugs with low bioavailability (oral heparin)
B. Nanoparticles for the selective drug targeting (inflammatory bowel disease)
C. Nanocapsules for intracellular drug therapy (cancer)
drug intestinal barrier organs cells, receptors
B CA
Nanoparticles fororal delivery of macromolecules
Objective:
enhanced absorption of the drugtransport across biological barrier
avoidance of repetitive injections
polymeric nanoparticles and theirtherapeutic applications
Heparin
- anionic glycosaminoglycans- treatment and prophylaxis
of venous thrombosis
Nanoparticles for oral absorption
• parenteral administration• no oral absorption
do not pass the intestinal barrier
NANOPARTICLES:
• drug entrapment (protection)• emulsion techniques• biocompatible polymers • diameter: 300-500nm
FormulationsFormulationsCmax Cmax
(s)(s)Tmax (h)Tmax (h)
AUC/KgAUC/Kg
(s.h/Kg)(s.h/Kg)
Absolute FAbsolute F
(%)(%)
RL/PCLRL/PCL 20-2720-27 66 28.3 ± 8.228.3 ± 8.2 42.1 ± 12.342.1 ± 12.3
RS/RL/PLGARS/RL/PLGA 20-2420-24 4-54-5 29.4 ± 12.629.4 ± 12.6 43.9 ± 18.843.9 ± 18.8
RS/PLGARS/PLGA 22-2722-27 6-76-7 21.9 ± 18.421.9 ± 18.4 32.7 ± 27.432.7 ± 27.4
RLRL 19-2719-27 3-63-6 26.3 ± 12.326.3 ± 12.3 39.2 ± 18.339.2 ± 18.3
IV UFH solutionIV UFH solution NDND NDND 67.1 ± 29.267.1 ± 29.2 100100
Pharmacokinetic parameters after oral administration of UFH-loaded NP (2000 IU) in rabbits (n=6)
Nanoparticles for oral absorption
Cell culture model for adsorption studies:
1. Absorption barrier (cells)
2. Mucus ???
no influence on absorption kinetics of small drugsbut macromolecules ???
adhesion studies on the cells, but adhesion to the mucus ???
mucoadhesion is the starting point!!!
Nanoparticles for oral absorption
0.01 0.1 1 100
500
1000
1500 blank RS-NP, x0=0.47
RS/PLG-NP, x0=1.66
PLG-NP, x0= /
resp
onse
[arc
sec
onds
]
NP conc. [mgpolymer
/ml]
0.01 0.1 1 100
50
100
150
+ LMWH RS-NP, x0=1.14
RS/PLG-NP, x0=1.49
PLG-NP, x0= /
resp
onse
[arc
sec
onds
]
NP conc. [mgpolymer
/ml]
resonant mirror system:NP adhesion differs with surface charge
Nanoparticles for oral absorption
Aqueous boundary layer (ca. 40 µm)
Epithelialcells
Basolateral compartment
Mucuslayer
• mucin coated Caco-2 cells• confluent but not homogeneous mucin layer• stained with astra blue
Nanoparticles for oral absorption
1.563 0.781 0.391 0.195 0.0980
5
10
15
20
254h - mucin
bin
din
g [
%]
NP conc. [mg/ml]
RS+LMWH RS/PLGA+LMWH PLGA+LMWH LMWH
• cationic NP adhesion• LMWH adhesion correlates!!!
0.01 0.1 1 100
5
10
15
20 RS RS/PLGA PLGA RS+LMWH RS/PLGA+LMWH PLGA+LMWH
part
icle
adh
esio
n [%
]
NP conc. [mg/ml]
Nanoparticles for oral absorption
0.1 1 10
2
4
6
8
fluo 4h: 1.71±0.48%fluo 24h: 2.18±0.51%
fluorescein RS 4h RS/PLGA 4h PLGA 4h RS 24h RS/PLGA 24h PLGA 24h
% d
rug
tran
spor
ted
NP [mg/ml]0.1 1 10
2
4
6
8
LMWH 4h: 0.29±0.18%LMWH 24h: 0.40±0.09%
LMWH RS 4h RS/PLGA 4h PLGA 4h RS 24h RS/PLGA 24h PLGA 24h
% d
rug
tran
spor
ted
NP [mg/ml]
LMWH transport across mucin coated Caco-2 cell layers
Nanoparticles for oral absorption
Objective:
local drug delivery in IBD treatmenttargeting on tissue level
lower adverse effects by a selective drug release
Nanoparticle targetingin inflammatory bowel disease
Inflammatory bowel disease
• chronic, relapsing inflammation of the colon• unknown etiology• swelling and ulceration of mucosa and submucosa
therapeutic principles:• induce remission of outbreaks• prevent outbreaks during remission
drug treatmentoral (standard)a) rectal (limited use)b) intravenous (recent)
surgery ulcerative colitis Crohn’s disease
Histology of the inflamed colonic mucosa(TNBS-model for the rat)
healthy control colitis group increasesmucus production
colitis group withulcerated tissue
Nanoparticles in colitis
Localization of 100nm particles in the colon
TNBS intrarectal; 5 days particles orally
healthy control mucus layer ofcolitis group
ulcerated tissue ofcolitis group
Nanoparticles in colitis
• higher deposition in inflamed colon selectivity• size dependent accumulation• no influence on the deposition in other gut regions
0.1µm 1µm 10µm 0.1µm 1µm 10µm0
5
10
15
20
25
30
#
*
*
colitis groupcontrol group
part
icle
bin
ding
(%
)
0.1µm 1µm 10µm0
5
10
15
20
25
*
part
icle
bin
ding
[%]
particle size
Nanoparticles in colitis
0 2 4 6 8 10 12 14
0
1
2
3
TNBS
**
**
**
***
*
FK506
TN
BS
clin
ical
act
ivity
sco
reday
Nanoparticles in colitis
tacrolimus:(also FK-506 or Fujimycin)
• use in organ transplant • immunosuppressive• use in IBD
0
10
20
30
40
0.0000
0.0063
0.0125
0.0188
0.0250
***
*
B
***
**
**
TNBS-rectal
colo
n/b
od
y w
eig
ht
rati
o
MP
O a
ctiv
ity
[U/m
g t
issu
e]
healthy untretaed healthy FK506 colitis control NP control FK506 FK506-NP
0
10
20
30
40
0.0000
0.0063
0.0125
0.0188
0.0250
*
D
***
OXA-rectal
colo
n/b
od
y w
eig
ht
rati
o
MP
O a
ctiv
ity
[U/m
g t
issu
e]
healthy untreated healthy FK506 colitis control NP control FK506 FK506-NP
Nanoparticles in colitis
NP efficient after rectal administration
significant drug loss during intestinal transit
Nanoparticles in colitis
NP increase drug tissue penetration selectively
solution:
inflamed
healthy
NP:
inflamed
healthy
NP tissue penetration into inflamed tissue
into healthy tissue
CYP
P-gp
0
1
2
3
FK506 formulations
***
rel.
pene
trat
ion
inde
x[c
oliti
s/he
alth
y]
FK506 FK506+amio FK506+tro FK506-NP
Nanoparticles in colitis
Conclusion
Nanoparticles target the inflamed tissue in IBD
A) accumulate the drug in the inflamed tissue:
B) increase and selectivity of drug tissue penetration
I) high therapeutic efficiency
II) reduction of adverse effects
Nanocapsules in thetreatment of cancer
Objective:
local drug delivery of anticancer drugstargeting on cell level
higher efficiency by a selective drug release
Lipid nanocapsules and theirtherapeutic applications
• controlled drug release• controlled surfactant release (P-gp inhibitor) • slower release for higher drug loading
0 24 48 72 96 1200
20
40
60
80
100
0
200
400
600
800
1000
PE
G-H
S r
elea
se [µ
mol
]
etop
osid
e re
leas
e [%
]
time [h]
PEG-HS (LNC25) PEG-HS (LNC50) PEG-HS (LNC100) eto (LNC25) eto (LNC50) eto (LNC100)
Lipid nanocapsule preparation
basal activity
vinblastine
verapamil
2.5µmol
25nmol
1.09µmol
10.9nmol
0.63µmol
6.3nmol
0.0
0.5
1.0
1.5
2.0
*****
*
rela
tive
AT
Pas
e ac
tivity
of P
-gp
LNC25 LNC50 LNC100
in-vitro testing of LNC on glioma cell cultures:
Nanocapsules in cancer
• etoposide LNC up to 80-fold more potent• more effective in F98 cells
0.01 0.1 1 10 100 1000 100000
20
40
60
80
100
120
F98
viab
ility
[%]
etoposide [µmol]
eto LNC25 eto LNC50 eto LNC100 eto sol. eto + PEG-HS sol. blank LNC25 blank LNC50 blank LNC100
0.01 0.1 1 10 100 1000 100000
20
40
60
80
100
120
9L
viab
ility
[%]
etoposide [µmol]
eto LNC25 eto LNC50 eto LNC100 eto sol. eto + PEG-HS sol. blank LNC25 blank LNC50 blank LNC100
Confocal laser scanning microscopy
• LNC uptake in F98 (a) and 9L (b) cells• etoposide + blank LNC no effect
cellular uptake required !!!
Nanocapsules in cancer
Nanocapsules in cancer
LNC - an promising concept in cancer treatment ?
- intracellular uptake - drug release - PEG-HS release- potentially blocking the P-gp
overcoming the multidrug resistance ???
Acknowledgments
Nancy
P. MaincentN. Ubrich
Y. Meissner
Gifu
Y. KawashimaH. TakeuchiYamamoto
Saarbrücken
C.M. LehrU. Schäfer
D. NeumannT. Betz
Angers
J.P. BenoîtF. Lacoeuille
E. Garcion