Beruflich Dokumente
Kultur Dokumente
www.elsevier.com/locate/addr
Abstract
Thiolated polymers or designated thiomers are mucoadhesive basis polymers, which display thiol bearing side chains. Based
on thiol/disulfide exchange reactions and/or a simple oxidation process disulfide bonds are formed between such polymers and
cysteine-rich subdomains of mucus glycoproteins building up the mucus gel layer. Thiomers mimic therefore the natural
mechanism of secreted mucus glycoproteins, which are also covalently anchored in the mucus layer by the formation of
disulfide bondsthe bridging structure most commonly encountered in biological systems. So far the cationic thiomers
chitosancysteine, chitosanthiobutylamidine as well as chitosanthioglycolic acid and the anionic thiomers poly(acylic acid)
cysteine, poly(acrylic acid)cysteamine, carboxy-methylcellulosecysteine and alginatecysteine have been generated. Due to
the immobilization of thiol groups on mucoadhesive basis polymers, their mucoadhesive properties are 2- up to 140-fold
improved. The higher efficacy of this new generation of mucoadhesive polymers in comparison to the corresponding
unmodified mucoadhesive basis polymers could be verified via various in vivo studies on various mucosal membranes in
different animal species and in humans. The development of first commercial available products comprising thiomers is in
progress. Within this review an overview of the mechanism of adhesion and the design of thiomers as well as delivery systems
comprising thiomers and their in vivo performance is provided.
D 2005 Elsevier B.V. All rights reserved.
Keywords: Mucoadhesion; Thiolated polymers; Thiomers; Disulfide bonds; Thiolated poly(acrylic acid); Thiolated chitosan
Contents
1.
2.
Introduction . . . . . . .
Synthesis of thiomers . .
2.1. Cationic thiomers
2.2. Anionic thiomers
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
1570
1571
1571
1571
B
This review is part of the Advanced Drug Delivery Reviews theme issue on bMucoadhesive Polymers: Strategies, Achievements and
Future ChallengesQ, Vol. 57/11, 2005.
* Tel.: +43 512 507 5371; fax: +43 512 507 2933.
E-mail address: andreas.bernkop@uibk.ac.at.
0169-409X/$ - see front matter D 2005 Elsevier B.V. All rights reserved.
doi:10.1016/j.addr.2005.07.002
1570
3.
1. Introduction
Since the concept of mucoadhesion has been pioneered in the 1980s, numerous attempts have been
undertaken in order to improve the adhesive properties of polymers. These attempts include approaches
such as the use of linear poly(ethylene glycol) as
adhesion promoter for hydrogels [1], the neutralization of ionic polymers [2], mucoadhesion by a sustained hydration process [3] and the development of
polymeradhesin conjugates (e.g. [4,5]) providing a
specific binding to epithelia. However, all these systems are based on the formation of non-covalent
bonds such as hydrogen bonds, van der Waals forces,
and ionic interactions. Accordingly, they provide only
relative weak mucoadhesion, in many cases insufficient to guarantee the localization of a drug delivery
system at a given target site. Mucoadhesive polymers
have therefore in many cases not proven to be effective as dpharmaceutical glueT [6,7].
A presumptive new generation of mucoadhesive
polymers are thiolated polymersdesignated thiomers [8]. In contrast to well-established mucoadhesive
polymers these novel polymers are capable of forming
covalent bonds. The bridging structure most commonly encountered in biological systemsthe disulfide bondhas thereby been discovered for the
covalent adhesion of polymers to the mucus gel
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
1571
1571
1573
1574
1575
1575
1575
1576
1576
1577
1577
1577
1577
1578
1578
1579
1580
1580
Polymer
+
SH
thiol bearing
ligand
SH
SH
SH
SH
SH
SH
Thiomer
SH
Thiomer
SH
Thiomer
S-S
Thiomer
SH
Mucin
S-S
Mucin
HS
Mucin
Mucin
Mucin
SH
Ox.
Thiomer
S-S
Mucin
review the so far gained knowledge on the mucoadhesive properties of thiomers is summarized and discussed. The provided information should represent a
good starting point for further developments in this
promising research field.
2. Synthesis of thiomers
2.1. Cationic thiomers
Cationic thiomers are mainly based on chitosans.
The primary amino group at the 2-position of the
glucosamine subunits of this polymer is the main
target for the immobilization of thiol groups. As outlined in Fig. 3 sulfhydryl bearing agents can be covalently attached to this primary amino group via the
formation of amide or amidine bonds. In case of the
formation of amide bonds the carboxylic acid group
of the ligands cysteine and thioglycolic acid reacts
with the primary amino group of chitosan mediated
for instance by carbodiimides [1012]. An unintended
oxidation of thiol groups during synthesis can be
avoided by performing the reaction under inert conditions. Alternatively the synthesis can be performed
1571
1572
CH 2 OH
OH
HO
OH
R2
OH
NH
O
NH
NH 2
O
SH
HO
SH
OH
Chitosan-Cysteine [10]
Alginate-Cysteine [15]
CH 2OH
O
CH2 OH
OH
NH
H
O
OH
OH
H
OH
H
SH
O
H
O
CH2 OH
COOH
NH
SH
O
SH
Carboxymethycellulose-Cysteine [16,17]
NH2+ClH
NH
H
OH
H
CONH
COOH
COOH
HS
COOH
CH2OH
Chitosan-Thiobutylamidine [13,14]
O
CH2OH
HO
CH2OH
H
H
O
H
H
O
H
OH
H
OH
O
H
OH
H
O
H
H
OH
CH3
H
HO
OH
CONH
OH
n
O
R2 =
R1 = OH
OH
SH
COOH
SH
N
H
CH3
COOH
CH3
COOH
O
CONH
COOH
NH
COOH
COOH
HS
COOH
HS
no mucosal surfaces in which mucins with cysteinerich subdomains are not present. In contrast to noncovalent bonds disulfide bonds are not influenced by
factors such as ionic strength and pH. Velocity and
extent of disulfide bond formation depends on the
concentration of thiolate anions representing the reactive form for thiol/disulfide exchange reactions and
oxidation processes. The concentration of thiolate
anions in turn depends on:
! the pK a value of the thiol group. In dependence on
the polymer backbone and the chemical structure
of the ligand, more or less reactive thiomers can be
designed. Thiol groups of the chitosanthiobutylamidine conjugate (Fig. 3), for instance, exhibit a
pK a value of 9.9 [13], whereas the pK a value of the
thiol groups of poly(acrylate)cysteine conjugates
is 8.35 [24];
! the pH of the thiomer. As only ionic thiomers are
used, they all display a high buffer capacity. The
buffer capacity of a sodium poly(acrylate) matrix
tablet, for instance, can be compared with that of an
at least 25 M acetate buffer. As all charged groups
remain concentrated on the polymeric network a
kind of dmicroclimateT can be established [25].
The reactivity of thiol groups can consequently be
controlled by adjusting the pH of the polymer to a
certain level. The higher the pH is adjusted, the more
reactive are the thiol groups and vice versa; and
! the pH of the surrounding medium. The reactivity of
thiol groups inside the polymeric network is mainly
controlled by the pH of the thiomer, whereas the
reactivity on the surface of the polymer is more
controlled by the pH of the surrounding medium.
As the mucus gel layer being close to the epithelium
has a pH around 7, thiol groups penetrating into the
mucus are always sufficiently reactive.
1573
disulfide bond breaker dithiothreitol already immobilized mucin could be completely removed from the
thiolated polymer [8].
3.2. In situ cross-linking process
Another likely mechanism being responsible for
the improved mucoadhesive properties of thiomers is
based on their in situ cross-linking properties. During
and after the interpenetration process, which could
be verified for mucoadhesive polymers such as
poly(acrylic acid) recently [26], disulfide bonds are
formed within the thiomer itself leading to additional
anchors via chaining up with the mucus gel layer.
The mechanism is illustrated in Fig. 4. It is similar to
Chain Links:
Polymer
Interpenetration
In situ cross-linking
SS
SS
Fig. 4. Schematic presentation of improved mucoadhesion by an insitu cross-linking in comparison to chain links.
1574
1,E+03
G' (Pa)
1,E+02
1,E+01
1,E+00
1,E-01
0
time (h)
Fig. 5. Increase in the elastic properties ( GV) of a 1.5% (m/v)
chitosanTBA conjugate gel at pH 5.5 and 37 8C as a function of
time. Indicated values are means (FS.D.) of at least three experiments (adopted from Bernkop-Schnurch et al. [13]).
4. Mucoadhesive properties
The mucoadhesive properties of thiomers in comparison to well-established polymers are discussed in
detail in this issue by Grabovac et al. Due to the
immobilization of thiol groups on all so far tested
polymers, their mucoadhesive properties were significantly improved irrespectively from the evaluation
method. In case of anionic mucoadhesive polymers
the poly(acrylic acid)cysteine conjugate seems to be
a good example for this observation. Marschutz et al.
could show that the viscosity of poly(acrylic acid)/
mucin mixturesdirectly correlating with the interactions of the polymer with the mucus and consequently
indicating the mucoadhesive propertiescan be more
than 10-fold improved [28]. The same thiolated polymer showed in comparison to the corresponding
unmodified polymer more than 2-fold and 20-fold
improved mucoadhesive properties in tensile studies
and by using the rotating cylinder method, respectively [28]. In addition, it could be shown that the
residence time of poly(acrylic acid) microparticles on
the small intestinal mucosa can be more than 3-fold
prolonged by the immobilization of thiol groups [29].
In case of cationic thiomers, on the other hand, the
chitosanthiobutylamidine conjugate seems to be a
good example, as it has been evaluated by various
mucoadhesion test system. We demonstrated a more
than 100-fold increased viscosity of chitosanthiobutylamidine conjugate in comparison to unmodified
chitosan [13]. Moreover in tensile studies and rotating
cylinder studies the mucoadhesive properties of the
thiolated version were 100-fold and 140-fold
improved, respectively [13,14].
In case of both mentioned thiomers the molecular
mass of the polymer chains had a great impact on their
mucoadhesive properties. For the anionic as well as
for the cationic thiomer the highest mucoadhesive
properties were achieved when they exhibited a medium molecular mass. In case of poly(acrylic acid)
1575
being based on thiolated polymers do not disintegrate. Because of the formation of disulfide bonds
within the polymeric network, the particles are stabilized [29,36]. Consequently, also a controlled drug
release out of thiomer micro- and nanoparticles can
be provided.
Recently, microparticles comprising poly(acrylate)cysteine were generated via the solvent evaporation emulsification method. Particles as illustrated in
Fig. 6 were of spherically and partially porous structure and had a main size in the range of 2060 Am
with a center at 35 Am. Because of the formation of
disulfide bonds within the particles they did not disintegrate under physiological conditions within 48 h.
In addition, a controlled drug release of a model
peptide drug was achieved. Due to the immobilization
of thiol groups on poly(acrylic acid) the mucoadhesive properties of the corresponding microparticles
were 3-fold improved [36].
5.2. Matrix tablets
S
HS
S
S
S
SH
HS
Acc.V Spot Magn WD
20.0 kV 4.0 2567x 17.7
10 m
1576
whereas tablets being based on thiolated polycarbophil did not disintegrable at all [17]. Moreover even
no erosion of this swollen drug carrier matrix could
be observed within an observation period of 24 h.
When attached in dry form the mucoadhesion of
matrix tablets is additionally improved by an augmented interpenetration process depending on the
swelling behavior of the delivery system. In order
to make use of this simple adhesion by hydration
process also in case of peroral delivery, matrix
tablets can be enteric coated. If an adhesion shall
be achieved already in the stomach, the coating with
a triglyceride seems to be sufficient in order to avoid
an unintended adhesion in the oral cavity or oesophagus [38]. Additionally, matrix tablets comprising
a thiomer offer the advantage that a controlled drug
release can be easily achieved out of this type of
mucoadhesive dosage forms, which could already be
demonstrated for numerous drugs (e.g. 37,39 40 41).
Hornof et al., for instance, could show that by
simple homogenizing the thiomer with the drug of
choice and compressing tablets out of it results in
many cases in delivery systems, which can guarantee even a zero order release profile for several
hours [41]. The drug release rate is thereby predominately controlled by a hydration and diffusion
process.
5.3. Gels
Mucoadhesive gels are useful in case of intraoral,
vaginal, nasal and ocular delivery. So far, however,
mucoadhesive gel formulations have not reached their
full potential, as the adhesive properties of such delivery systems are often insufficient. The great advantage of the use of thiomers in gel formulations has to be
seen not only in their mucoadhesive but also in their in
situ gelling properties [13,19]. Strong mucoadhesive
properties are senseless, if the adhesive bond fails
rather within the gel formulation itself than between
the gel and the mucosa. Due to the in situ gelling
properties of thiomers, however, this shortcoming can
be overcome.
5.4. Liquid formulations
Thiomers were shown to be stable when stored in
dry form [42]. In aqueous solutions, however, they
were shown to form disulfide bonds in a pH-dependent manner. Because of this instability in aqueous
solutions thiomers have so far not been used in liquid
formulations. Recently, however, Hornof could
demonstrate that thiomers can even be stabilized in
aqueous solutions when the liquid formulations are
produced under inert conditions and the vessels are
packed in an aluminium foil containing an oxygen
scavenger such as iron-oxides inside [43]. Based on
this technology first mucoadhesive liquid formulations comprising thiomers were prepared and tested
in vivo.
In particular in the ophthalmic field thiomers have
already shown potential in form of liquid formulations. In case of the dry eye syndrome the most
prevalent disease in the eye, for instance, liquid
thiomer formulations might be highly beneficial.
One of the most important reasons for this disease
seems to be a defective mucus layer on the ocular
surface. Mucus acts as surfactant, and is therefore
important for the wettability of the epithelial surface
[44]. The main treatment for this disease is the use
of tear substitutes. Most of these formulations contain mucoadhesive hydrophilic polymers such as
carbomer or sodium hyaluronate. The mucoadhesive
properties of these polymers, however, are quite
insufficient making a frequent instillation necessary.
Because of their ability to interact with cysteine-rich
subdomains of mucus glycoproteins on the ocular
surface, eye drops containing a thiomer should be
able to prolong the stability of the precorneal tear
film for a comparable longer time period. A parameter to characterize the quality of the tear film is
the tear film break-up time, which is defined as the
time period after a blink in which the tear film
becomes unstable and dry spots evolve on the cornea. Normally the break-up time exceeds the timespan between blinks, but in patients with dry eye
syndrome the break-up time is decreased to less than
5 s. In Fig. 7 the comparison of the effect of a wellestablished commercial product containing carbomer
and a formulation containing 0.2% (m/v) polyacrylic
acidcysteine and mannitol as tonicity agent on the
tear film break-up time is shown in human volunteers. The eye drops containing thiolated polyacrylic
acid had a positive effect on the tear film stability,
whereas no difference in the tear film break-up time
was observed after application of an isotonic manni-
BUT [sec]
4
3
2
1
0
12
0 min
5 min
Time of determination
Fig. 7. Effect of different eye drop formulations on the tear film
break-up time (BUT) of healthy volunteers. The BUT was measured
before eye drop instillation (0 min) and 5 min after eye drop
application in a single eye. The formulations tested were aqueous
eye drops containing an isotonic mannitol solution (grey bars), a
commercially available formulation containing carbomer (white
bars) and aqueous eye drops containing 0.2% (m/v) poly(acrylic
acid)cysteine conjugate and mannitol as tonicity agent (black bars)
(adopted from Hornof [43]). Indicated values are means of at least 5
experiments.
1577
0,6
0,5
heparin (IU/ml)
0,4
0,3
0,2
0,1
0
8 10 12 14 16 18 20 22 24
time [hours]
Fig. 8. Comparison of the concentration profiles of low molecular
weight heparin in plasma obtained after peroral administration of
low molecular heparin incorporated in minitablets comprising thiolated poly(acrylatic acid) (o) and in minitablets comprising the
corresponding unmodified polymer (.) in rats. Data represent the
mean F S.D. of five experiments [45].
1578
120
110
100
90
80
70
60
50
40
0
12
16
20
24
28
32
Time (hours)
Fig. 9. Glycemic profiles in diabetic mice after single oral administration of pegylated insulin loaded minitablets comprising thiolated poly(acrylic acid) (.) and of a pegylated insulin solution (o).
Each point represents the mean F S.D. of 10 experiments (adopted
from Caliceti et al. [47]).
Recently, the potential of a thiomer gel formulation could be demonstrated by in vivo studies. Leitner et al. developed a nasal gel formulation for
systemic delivery of hGH. The efficacy of a mucoadhesive gel formulation being based on unmodified
polycarbophil and polycarbophilcysteine was compared in rats. Results as shown in Fig. 11 demon-
1579
100
95
85
0
12
16
20
24
Time [h]
Fig. 10. Decrease in plasma calcium level as a biological response
for the salmon calcitonin bioavailability in fasted rats after oral
administration of thiolated chitosan minitablets targeted to the
small intestine (E), of unmodified chitosan minitablets targeted to
the small intestine (o), of thiolated chitosan minitablets targeted to
the stomach (*), and of unmodified chitosan minitablets targeted to
the stomach (n). Indicated values are the mean results of five
experiments (adopted from Guggi et al. [38,53]).
140
120
100
80
60
40
20
0
0
time (h)
Fig. 11. Concentrationtime profiles of human growth hormone
(hGH) in rat plasma obtained after nasal administration of hGH
incorporated in a thiolated polyacrylate gel (.) and in the corresponding unmodified polyacrylate gel (o). Data represent the
mean F S.D. of 45 experiments (adopted from Leitner et al. [54]).
1580
350
7. Conclusion
fluorescein [ng/ml]
300
250
200
150
100
50
0
0 1 2 3 4 5 6 7 8
time [h]
Fig. 12. Fluorescein concentration in the cornea/tearfilm compartment of a volunteer after application of a thiolated polyacrylate
insert (o) and the corresponding unmodified polyacrylate insert (.)
containing 15% fluorescein as model drug (adopted from Hornof et
al. [41]).
tion of the device on the ocular surface and a controlled release. Dosing of the drugs is also more
accurate and the risk of systemic side-effects is
decreased. Nevertheless, despite all these advantages
ocular inserts were so far not widely used in ocular
therapy. Inserts without appropriate mucoadhesive
properties can move around on the ocular surface
causing irritation and can be easily lost. The erosion
and/or disintegration into smaller pieces of soluble
inserts results in occasional blurring of vision. Ocular
inserts comprising thiomers, however, do not have
these disadvantages.
Hornof et al., for instance, could shown that inserts
based on thiolated poly(acrylic acid) were not soluble
and had good cohesive properties. In addition, a controlled release was achieved for the incorporated
model drug sodium fluorescein. In vivo studies in
human volunteers showed that inserts based on thiolated poly(acrylic acid) provide a fluorescein concentration on the ocular surface for more than 8 h, while
the fluorescein concentration rapidly decreased after
application of aqueous eye drops or inserts based on
unmodified poly(acrylic acid). A representative result
of this study is shown in Fig. 12 [41]. Moreover, these
inserts were well accepted by the volunteers. The
study indicated that ocular inserts based on thiolated
poly(acrylic acid) are promising new solid devices for
ocular drug delivery [58].
References
[1] J.J. Sahlin, N.A. Peppas, Enhanced hydrogel adhesion by
polymer interdiffusion: use of linear poly(ethylene glycol) as
an adhesion promoter, J. Biomater. Sci., Polym. Ed. 8 (1997)
421 436.
[2] J.D. Smart, I.W. Kellaway, H.E.C. Worthington, An in-vitro
investigation of mucosa-adhesive materials for use in controlled
drug delivery, J. Pharm. Pharmacol. 36 (1984) 295 299.
[3] W.J. Bologna, H.L. Levine, Ph. Cartier, D. de Ziegler,
Extended release buccal bioadhesive tablets, WO00/10536
(1998).
[4] B. Naisbett, J. Woodley, The potential use of tomato lectin for
oral drug delivery. Lectin binding to rat small intestine in vitro,
Int. J. Pharm. 107 (1994) 223 230.
[5] A. Bernkop-Schnurch, F. Gabor, M.P. Szostak, W. Lubitz, An
adhesive drug delivery system based on K99-fimbriae, Eur. J.
Pharm. Sci. 3 (1995) 293 299.
[6] R. Khosla, S.S. Davis, The effect of polycarbophil on the
gastric emptying of pellets, J. Pharm. Pharmacol. 39 (1987)
47 49.
[7] C.-M. Lehr, Bioadhesion Technologies for the delivery of
peptide and protein drugs to the gastrointestinal tract, Crit.
Rev. Ther. Drug 11 (1994) 119 160.
[8] A. Bernkop-Schnurch, V. Schwarz, S. Steininger, Polymers
with thiol groups: a new generation of mucoadhesive polymers? Pharm. Res. 16 (1999) 876 881.
[9] V.M. Leitner, G.F. Walker, A. Bernkop-Schnurch, Thiolated
polymers: evidence for the formation of disulphide bonds with
mucus glycoproteins, Eur. J. Pharm. Biopharm. 56 (2003)
207 214.
[10] A. Bernkop-Schnurch, U.M. Brandt, A.E. Clausen, Synthesis
and in vitro evaluation of chitosancysteine conjugates, Sci.
Pharm. 67 (1999) 196 208.
[11] A. Bernkop-Schnurch, T.E. Hopf, Synthesis and in vitro evaluation of chitosanthioglycolic acid conjugates, Sci. Pharm.
69 (2001) 109 118.
[12] C.E. Kast, A. Bernkop-Schnurch, Thiolated polymersthiomers: development and in vitro evaluation of chitosanthioglycolic acid conjugates, Biomaterials 22 (2001) 2345 2352.
1581
1582
[53] D. Guggi, C.E. Kast, A. Bernkop-Schnurch, In vivo evaluation of an oral salmon calcitonin-delivery system based on a
thiolated chitosan carrier matrix, Pharm. Res. 20 (2003)
1989 1994.
[54] V. Leitner, D. Guggi, A. Bernkop-Schnurch, Thiomers in noninvasive peptide delivery: in vitro and in vivo characterisation
of a polycarbophilcysteine/glutathione gel formulation for
hGH, J. Pharm. Sci. 93 (2004) 1682 1691.
[55] V.M. Leitner, D. Guggi, A. Bernkop-Schnurch, Design and in
vivo evaluation of a nasal delivery system for hGH, 5th Central
European Symposium on Pharmaceutical Technology and Biotechnology, Faculty of Pharmacy, Ljubljana, Slovenia, 2003.
[56] C. Baudoin, Side effects of antiglaucomatous drugs on the
ocular surface, Curr. Opin. Ophthalmol. 7 (1996) 80 86.
[57] A. Topalkara, C. Gurtler, D.S. Arici, M.K. Arici, Adverse
effects of topical antiglaucoma drugs on the ocular surface,
Clin. Exp. Ophthalmol. 28 (2000) 113 117.
[58] J. Weidener, Mucoadhesive ocular inserts as an improved
delivery vehicle for ophthalmic indications, Drug Discov.
Today 8 (2003) 906 907.