Development of a mucoadhesive nanoparticulate drug ... · -development of a mucoadhesive...
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Development of a mucoadhesive nanoparticulate drug delivery system for a targeted drug release in the bladder Jan Barthelmes Department of Pharmaceutical Technology
Transcript of Development of a mucoadhesive nanoparticulate drug ... · -development of a mucoadhesive...
Development of a mucoadhesive nanoparticulate drug delivery system for a targeted drug release
in the bladder
Jan Barthelmes
Department of Pharmaceutical Technology
1.1. IntroductionIntroduction
2.2. Methods & ResultsMethods & Results
3.3. ConclusionConclusion
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ContentContent
IntroductionIntroduction
− treated by oral administration of pharmaceutical compounds���� systemic delivery
− Intravesical Drug Delivery (IDD)
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Diseases of the urinary bladderDiseases of the urinary bladder
cancer inflammation, infection incontinence
− periodical voiding of urine dilutes and washes out the drug� reduces the residence time of drug and lead to a new
administration� repeated catheterizations increase potential for infections− very low permeability of the urothelium in the diseased state
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Intravesical drug delivery (IDD) limitationsIntravesical drug delivery (IDD) limitations
IntroductionIntroduction
− development of a mucoadhesive nanoparticulate drug delivery system for local use in intravesical therapy
� retarding release of the drug� prolong the residence time of the drug in the bladder
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Purpose of the present studyPurpose of the present study
IntroductionIntroduction
− trimethoprim (TMP) was used as an effective local therapy from cystitis in the bladder
− fluorescein diacetate (FDA) wasused as fluorescent marker
1. Introduction
2.2. Methods & ResultsMethods & Results
3. Conclusion
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ContentContent
Methods & ResultsMethods & Results
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Preparation of the matrix of the drug delivery systemPreparation of the matrix of the drug delivery system
Chitosan - Thioglycolic acid (TGA)
Thioglycolic acid
Chitosan
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Preparation of the drug delivery systemPreparation of the drug delivery system
Methods & ResultsMethods & Results
Chitosan or chitosan-TGA in 0.01 M acetic acid /sodium acetat buffer 0.5% (w/v) pH 6.2
TPP solution in demineralised water 0.2% (w/v)
Nanoparticles (NP)
O
H
NH3+H
OH
CH2OH
H
O
H
NH H
OH
CH2OH
H
O O
OHHN
H OH CH2OHH
O H NH3+
HHOHOH2C H
O
O
O
H
NH3+
H
HO
HOH2C
H
O
H
NHH
OH
CH2OH
H
O
O
O H NH
HHOHOH2C H
OH+H3N
H OH CH2OHH
O
O
-O
PO
PO
PO-
O
O
O
HO
HO
HO
O
SH
O
HS
OHS
O
SH
O
O
O
H
NH3+H
OH
CH2OH
H
O
H
NH H
OH
CH2OH
H
O O
OHHN
H OH CH2OHH
O H NH3+
HHOHOH2C H
O
O
O
H
NH3+
H
HO
HOH2C
H
O
H
NHH
OH
CH2OH
H
O
O
O H NH
HHO
HOH2CH
OH+H3N
H OH CH2OHH
O
O
-O
PO
PO
PO-
O
O
O
HO
HO
HO
O
SH
O
HS
OHS
O
SH
O
O
O
H
NH3+H
OH
CH2OH
H
O
H
NH H
OH
CH2OH
H
O O
OHHN
H OH CH2OHH
O H NH3+
HHOHOH2C H
O
O
O
H
NH3+
H
HO
HOH2C
H
O
H
NHH
OH
CH2OH
H
O
O
O H NH
HHOHOH2C H
OH+H3N
H OH CH2OHH
O
O
-O
PO
PO
PO-
O
O
O
HO
HO
HO
O
SH
O
HS
OHS
O
SH
O
O
O
H
NH3+H
OH
CH2OH
H
O
H
NH H
OH
CH2OH
H
O O
OHHN
H OH CH2OHH
O H NH3+
HHOHOH2C H
O
O
O
H
NH 3+
H
HO
HOH2C
H
O
H
NHH
OH
CH 2OH
H
O
O
O HNH
HHO
HOH2C H
OH+H3N
H OH CH2OHH
O
O
-O
PO
PO
PO-
O
O
O
HO
HO
HO
O
SH
O
HS
OHS
O
SH
O
O
O
H
NH3+H
OH
CH2OH
H
O
H
NH H
OH
CH2OH
H
O O
OHHN
H OH CH2OHH
O H NH3+
HHOHOH2C H
O
O
O
H
NH 3+
H
HO
HOH2C
H
O
H
NHH
OH
CH 2OH
H
O
O
O HNH
HHO
HOH2C H
OH+H3N
H OH CH2OHH
O
O
-O
PO
PO
PO-
O
O
O
HO
HO
HO
O
SH
O
HS
OHS
O
SH
O
O
O
H
NH3+H
OH
CH2OH
H
O
H
NH H
OH
CH2OH
H
O O
OHHN
H OH CH2OHH
O H NH3+
HHOHOH2C H
O
O
O
H
NH 3+
H
HO
HOH2C
H
O
H
NHH
OH
CH 2OH
H
O
O
O HNH
HHO
HOH2C H
OH+H3N
H OH CH2OHH
O
O
-O
PO
PO
PO-
O
O
O
HO
HO
HO
O
SH
O
HS
OHS
O
SH
O
O
O
H
NHH
OH
CH2 OH
H
O
H
NH3+ H
OH
CH2OH
H
O
H
NHH
OH
CH2OH
HO O
O
OHNH
H OH CH2OHH
O
O
O
O H NH
HHOHOH2C H
OH
+ H3N
H
OH
CH2OH
H
OH
NHH
HO
HOH2C
H
O
O
O
O
H
NH3+
H
HO
HOH2C
HO
O
O
O
O
O
SO
SH
S
O
H
+ H3 NH
OH
CH2OH
H
O
H
HN
H
OH
CH2OH
H
OS
SO
HNH
HOH
CH2OHH
OH
NH3+
H
HO
HOH2C
H
O
S
OS
O
H
NHH
OH
CH2 OH
H
O
H
NH3+ H
OH
CH2OH
H
O
H
NHH
OH
CH2OH
HO O
O
OHNH
H OH CH2OHH
O
O
O
O H NH
HHOHOH2C H
OH
+ H3N
H
OH
CH2OH
H
OH
HN
H
HO
HOH2C
H
O
O
O
O
H
NH3+
H
HO
HOH2C
HO
O
O
O
O
O
SH
O
SH
O
H
+ H3NH
OH
CH2OH
H
O
H
HN
H
OH
CH2OH
H
OS
SO
HNH
HOH
CH2OHH
OH
NH3+
H
HO
HOH2C
H
O
S
OS
SH
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Preparation of the drug delivery systemPreparation of the drug delivery system
Methods & ResultsMethods & Results
Different amounts of 0.5% (v/v) H2O2solution
Nanoparticles (NP)
O
H
NH3+H
OH
CH2OH
H
O
H
NH H
OH
CH2OH
H
O O
OHHN
H OH CH2OHH
O H NH3+
HHOHOH2C H
O
O
O
H
NH3+
H
HO
HOH2C
H
O
H
NHH
OH
CH2OH
H
O
O
O H NH
HHOHOH2C H
OH+H3N
H OH CH2OHH
O
O
O
SH
O
HS
OS
O
S
O
O
Methods & ResultsMethods & Results
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Characterization of the drug delivery system
Table 1. Mean particle diameter and zeta potential of chitosan-TGA nanoparticles obtained by ionic gelation with TPP and followed by different oxidation with H2O2, respectively. Indicated values are means � SD (n � 3).
NanoparticlesMean particle diameter [nm]
Polydispersity index
Zeta potential [mV]
Ionically crosslinked
Chitosan 266 � 64 0.44 7 � 1
Chitosan-TGA 197 � 24 0.38 7 � 1
Covalently crosslinked
Chitosan TGA (ox1) 183 � 7 0.31 13 � 3Chitosan TGA (ox2) 186 � 6 0.29 12 � 1
Methods & ResultsMethods & Results
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Characterization of the drug delivery system
Fig. 2. Size distribution of ionically crosslinked nanoparticles as wellas covalently crosslinked nanoparticles. Indicated values are means ± SD of last three experiments.
Methods & ResultsMethods & Results
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Characterization of the drug delivery system
Table 2.Amount of thiol groups and disulfide bonds immobilised on the basic thiomer Chitosan-TGA and nanoparticles after ionic gelation with TPP and different degrees of oxidation with H2O2, respectively. Indicated values are means � SD (n � 3).
H2O2
[µmol]-SH [µmol/g] -S-S- [µmol/g] �-SH [µmol/g]
Chitosan-TGA - 1456 136 1728 � 62
Chitosan-TGA NP - 1391 178 1747 � 36
Chitosan-TGA NP (ox1) 10.60 903 426 1753 � 55
Chitosan-TGA NP (ox2) 21.21 641 559 1758 � 27
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Characterization of the drug delivery system
Methods & ResultsMethods & Results
Fig. 3. Transmission electron microscopy images of the spherical shape of nanoparticles based on chitosan [A], chitosan-thioglycolic acid [B], chitosan-thioglycolic acid with 426 µmol/g disulfide bonds [C] and chitosan- thioglycolic acid with 559 µmol/g disulfide bonds [D]. Displayed bar represents 4.0 µm.
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Characterization of the drug delivery system
Methods & ResultsMethods & Results
Fig. 4. Payload of trimethoprim [white bars] and fluorescein diacetate [black bars] loaded nanoparticles based on chitosan, chitosan-thioglycolic acid and
chitosan- thioglycolic acid with 426 µmol/g (ox1) and 559 µmol/g (ox2) disulfide bonds. Indicated values are means ± SD (n � 3). * Differs from TMP, p < 0.05.
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In vitro mucoadhesion studies on porcine urinary bladdersMethods & ResultsMethods & Results
FDA loaded NP
Instillation of
8 mg prehydrated NP
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In vitro mucoadhesion studies on porcine urinary bladders
Methods & ResultsMethods & Results
Fig. 5. Percentage of fluorescein diacetate remaining on porcine urinary bladders as a function of time. Studies were carried out with chitosan-thioglycolic acid nanoparticles [black bars] and unmodified chitosan nanoparticles [white bars] as control. Indicated values are means ± SD (n � 3). * Differs from unmodified chitosan nanoparticles, p < 0.05.
− female Sprague-Dawley rats, average body weight 250 g − rats were fasted but had free access to water− anesthetized by an injection of ketamine (20 mg/kg)/xylazin-
hydrochloride (4 mg/kg) mixture− before urethral catheterization animals were positioned in supine
position, and micturition was induced through mild caudal abdominal massage
− 500 µl of each formulation was administered
Methods & ResultsMethods & Results
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In vivo evaluation of particles with rats
Animal grouping Administrated formulations
Group 1 FDA suspension
Group 2FDA loaded unmodified chitosan nanoparticles
Group 3FDA loaded chitosan-TGA nanoparticles
Methods & ResultsMethods & Results
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In vivo evaluation of particles with rats
Fig. 6. Amount of fluorescein diacetate remaining on rats bladders. Fluorescein diacetate was applied without any excipients [black bars] or incorporated in unmodified chitosan nanoparticles [grey bars] or chitosan-thioglycolic acid nanoparticles [white bars]. Indicated values are means ± SD (n � 3).
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Mucoadhesion on mucosa of urinary bladdersMethods & ResultsMethods & Results
O
H
NH 3+H
OH
CH 2OH
H
O
H
NH H
OH
C H2OH
H
O O
OHHN
H OH CH2OHH
O H NH 3+
HHOHOH2C H
O
O
O
H
NH3+
H
H O
HOH2C
H
O
H
NHH
OH
CH 2OH
H
O
O
O H NH
HHO
HOH2C H
OH+H3N
H OH CH 2OHH
O
O
O
SH
O
HS
OS
O
S
O
O
Layers in the bladder wall
Mechanism of disulfide bond formation between thiomers and mucus glycoproteins
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Release studiesRelease studies
Methods & ResultsMethods & Results
OXIDATION
H2O2
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TMP release studiesTMP release studies
Methods & ResultsMethods & Results
Fig. 7. Release properties of trimethoprim nanoparticles among simulated conditions with artificial urine as a function of crosslinking. Studies were carried out with nanoparticles based on chitosan [�], chitosan- thioglycolic acid [�], chitosan-thioglycolic acid with 426 µmol/g disulfide bonds [�] and 559 µmol/g [�] disulfide bonds. Indicated values are means ± SD (n � 3). Differs from chitosan nanoparticles, p < 0.05.
1. Introduction
2. Methods & Results
3.3. ConclusionConclusion
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ContentContent
ConclusionConclusion
− intravesical drug delivery system based on thiolated chitosan offers an adequate release profile besides its mucoadhesive properties
− chitosan-TGA NP showed:1. greater stability2. superior mucoadhesion3. more sustained and controlled release
� Finally, chitosan-TGA intravesical drug delivery system might be a useful tool for a local drug application in the urinary bladder, which allows:1. prolonged residence time at the target site2. enables sustained drug delivery of trimethoprim over a longer
time span
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THANK YOU FORTHANK YOU FORATTENTION!ATTENTION!
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