Vol.1, No.3, 127-133 (2009)
doi:10.4236/health.2009.13021
SciRes Copyright © 2009 Openly accessible at http://www.scirp.org/journal/HEALTH/
Health
Effects of structural modification of anti-inflammatory
steroidal antedrug on pro-inflammatory mediators and
inhibitory cytokines in human alveolar epithelial cells
Gui-Fang Wang1, Soonjo Kwon2*, Rakesh Sharma1, Hemang Patel2, Henry J. Lee3
1Chemical and Biomedical Engineering, Florida State University, Tallahassee, USA;
2Biological Engineering, Utah State University, Logan, USA
3Center for Anti-Inflammatory Research, College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Tallahassee,
USA
Received 23 September 2009; revised 19 October 2009; accepted 20 October 2009.
ABSTRACT
The anti-inflammatory effects of the new ster-
oidal antedrug, 21-acetyloxy-9α-fluoro-11β-hy-
droxyl-3, 20-dioxo-1, 4-pregnadieno-[16α, 17α-d]
isoxazoline (FP-ISO-21AC), on nitric oxide (NO)
and interleukin 8 (IL-8) production, were inves-
tigated together with its parent steroid predni-
solone (PRED). PRED is one of the anti-in-
flammatory steroids but has systemic side ef-
fects which limit the use of it. PRED was modi-
fied with ‘antedrug concept’ to create safer
drugs that attack problems such as inflamma-
tion, then quickly become inactive before they
can cause systemic side effect. We had a test
about the effect of the modified anti-inflamma-
tory steroidal antedrug on anti-inflammatory
activity. The present study evaluated their ability
to inhibit cytokine-induced NO and IL-8 produc-
tion in human alveolar epithelial cells. We also
investigated their ability to enhance the expres-
sion of inhibitory cytokine receptor, interleukin
22 receptor (IL-22R) in human alveolar epithelial
cells. Our results showed that FP-ISO-21AC sh-
owed higher ability to inhibit the cytokine - in-
duced production of NO than PRED. Exogenous
IL-22 was added to the media of both human
alveolar epithelial cells (A549) and human lung
fibroblast (HLF-1). In the presence of the ex-
ogenous inhibitory cytokine IL-22, further re-
duction of NO production was observed in A549
cells, which express IL-22R, but not in HLF1,
which does not express IL-22R. These data
suggested that the steroidal antedrugs en-
hanced the expression of IL-22R. FP-ISO- 21AC
showed higher potency than PRED to restore
the expression of IL-22R. FP-ISO-21AC further
reduced NO production to 27% and PRED
further reduced NO production to 39%. In con-
clusion, a synthesized steroidal antedrug FP-
ISO-21AC showed higher anti-inflammatory ef-
fects than PRED by inhibiting the expression of
pro-inflammatory mediator NO and stimulating
the expression of IL-22R.
Keywords: Steroidal Antedrug; NO Production;
IL-8 Production; Anti-Inflammatory Cytokine Receptor;
IL-22 Receptor
1. INTRODUCTION
Glucocorticoid is one of the most commonly and effec-
tively used drugs to relieve inflammation [1-4]. However,
these drugs display a number of serious systemic side
effects such as suppression on pituitary-adrenal axis and
on the immune system, aggravation of diabetes, hyper-
tension, retardation of growth in children and osteoporo-
sis. These systemic side effects limit the clinical use of
corticosteroids [5-9]. Therefore, the main focus of present
research is to modify the structure of corticosteroids by
reducing the systemic side effects without losing their
anti-inflammatory function. A novel strategy was em-
ployed by incorporating of metabolically labile functional
group onto the steroid chain structure based on the ‘ante-
drug concept’. As Lee et al. described in previous papers,
antedrug is defined as a compound which acts locally on
the target tissue and it is rapidly metabolized to an inac-
tive metabolite through enzymatic reaction upon entry
into the systemic circulation [10-13].
The new anti-inflammatory steroidal antedrug, 21-Ac-
etyloxy-9α-fluoro-11β-hydroxy-3, 20-dioxo-1, 4-pregna-
dieno-[16α,17α-d] isoxazoline (FP-ISO-21AC) with
C-16, 17-isoxazoline ring systems was synthesized and
its pharmaceutical activities were evaluated [14]. Their
study also showed that FP-ISO-21AC has high binding

G. F. Wang et al. / HEALTH 1 (2009) 127-133
SciRes Copyright © 2009 Openly accessible at http://www.scirp.org/journal/HEALTH/
128
affinities to the glucocorticoid receptor prepared from
liver cytosol and exhibited its enhanced inhibitory effects
on LPS- induced nitric oxide (NO) production in murine
macrophage cells (RAW 264.7)[14]. Although this new
steroidal antedrug showed more potential anti-inflamma-
tory effects, few main mechanisms by which multiple
pro-inflammatory pathways are switched off and anti-
inflammatory pathways are switched on in asthma fol-
lowing pretreatment of steroidal antedrugs have been
identified. In present time, multiple pro-inflammatory
mediators have been implicated in asthma. However, the
inhibitory mechanisms of the inflammatory process still
remain to be investigated. There is increasing evidence
that certain cytokines have anti-inflammatory or immu-
nomodulatory effects and that their secretion might be
defective in asthmatic patients. As a result, anti- inflam-
matory cytokines (inhibitory cytokines) can block the
inflammatory process or suppress the intensity of the
inflammatory cascade. The severity and the persistence
of asthma depend on the “balance” between the pro-
inflammatory cytokines and the anti-inflammatory cyto-
kines [15]. The expression of anti-inflammatory cyto-
kines can be restored by the treatment with steroids or
theophylline [16].
In this study, we had a test on the effect of the modi-
fied anti-inflammatory steroid (FP-ISO-21AC, isoxa-
zoline derivative, anti-inflammatory steroidal antedrugs)
on anti-inflammatory activities. The present study ad-
dresses the following fundamental questions: 1) Does
structural modification of anti-inflammatory steroidal
antedrugs increase the therapeutic index of potent corti-
costeroids with reducing their systemic side effects? 2)
Do these anti-inflammatory steroidal antedrugs enhance
or restore the expression of anti-inflammatory proteins
(e.g. inhibitory cytokines)? More specifically, the effect
of FP-ISO-21AC, an active synthetic derivative of anti-
inflammatory steroidal antedrug, was evaluated on re-
duced production of pro-inflammatory mediators (IL-8
and NO). Simultaneously, FP-ISO-21AC was evaluated
on enhanced expression of anti-inflammatory cytokine
receptor IL-22R in human alveolar epithelial cells. FP-
ISO-21AC showed higher potency to restore the expres-
sion of IL-22R and further reduced NO production, com-
pared with PRED. A synthesized steroidal antedrug FP-
ISO-21AC showed higher anti-inflammatory effects than
PRED by inhibiting the expression of pro-inflammatory
mediator, NO and stimulating the expression of the in-
hibitory cytokine receptor, IL-22R. The study has enor-
mous diagnostic implications in asthmatic patients and
designing novel synthetic anti-inflammatory steroids with
less systemic side effects.
2. MATERIALS AND METHODS
2.1. Chemicals and Reagents
Recombinant human interleukin-1 (IL-1), recombi-
Figure 1. Structure of antedrug FP-ISO-21AC.
nant human tumor necrosis factor-a (TNF-), recombi-
nant human interferon- (IFN-) and recombinant human
interleukin-22 (IL-22) were obtained from R&D systems,
Inc (Minneapolis, USA). New steroidal antedrug, 21-
Acetyloxy-9α-fluoro-11β-hydroxy-3, 20-dioxo-1, 4-pre-
gnadieno-[16α, 17α-d] isoxazoline (FP-ISO-21AC) was
synthesized as described previously [17-19]. The struc-
ture of FP-ISO-21AC is shown in Figure 1.
2.2. Cell Culture
Human alveolar epithelial cells, A549 (ATCC, CCL- 185)
and human lung fibroblast, HFL1 (ATCC, CCL- 183)
were obtained from ATCC (Manassas, VA, USA). The
cells were grown on Ham's F12K medium with 2 mM of
L-glutamine adjusted to contain 1.5g/L sodium bicar-
bonate, 90% (ATCC, Manassas, VA, USA); and supplied
with 10% fetal bovine serum (ATCC, Manassas, VA,
USA). The cells were plated onto 75 cm2 flasks and in-
cubated at 37 °C with 95% air and 5% CO2. Once the
cells reached confluence, cells were typsinized with a
solution containing 0.25% (w/v) trypsin and 0.038%
(w/v) EDTA-4Na (Invitrogen, Carlsbad, California,
USA). Cells were then transferred to 24-well plates at
the seeding density of 1 X 104 per well. When the cells
reached confluence (approximately 7 days), they were
washed twice with PBS (Phosphate Buffered Saline).
2.3. Stimulation with Cytokines and/or
Steroidal Antedrugs
Once confluent, cells were incubated in serum-free me-
dia for 24 h before exposure to cytokine(s) and/or ster-
oidal antedrugs. Cells were then pre-incubated for 1 h
with PRED and FP-ISO-AC at 1M in cell culture me-
dium without serum. After washing cell layers with PBS
twice, cells were stimulated with Cytomix (IL-1, TNF-
 and IFN-) at 50 ng/ml each for 23 h at 37°C in cell
culture medium in combination with anti-inflammatory
cytokine (IL-22) at 20 ng/ml. Following cytokine expo-
sure, serum was returned to the culture media. NO and
IL-8 was analyzed at 24 h following stimulation.
2.4. Measurement of Nitric Oxide (NO)
NO production was measured by NO Analyzer (inNO,

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129
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Nitric Oxide Measuring System, Harvard apparatus,
Holliston, Massachusetts, USA). NO sensors were cov-
ered with a series of permeable membranes allowing
only NO to diffuse through from the sample solution and
react at the electrode surface, where it is oxidized. The
exchange of electrons resulted in an electrical current
which depends on the concentration of NO. The concen-
tration of NO in the sample is proportional to the amount
of NO that diffused through the membranes. Conse-
quently, the electrical current is proportional to the con-
centration of NO in the sample. The NO data were also
confirmed by Griess reagent system (Promega Corpora-
tion, WI).
2.5. Measurement of IL-8 Expression
IL-8 released into culture medium was measured with an
enzyme-linked immunosorbent assay (ELISA) sandwich
kit (ELISA quantikine, ref. D8050, R&D Systems, Inc,
Minneapolis, USA). Colorimetric changes were meas-
ured in a microplate reader (BioRad) at 490 nm. The IL-
8 level was estimated by interpolation from the calibra-
tion curve obtained using the standard IL-8.
3. RESULTS
PRED was structurally modified with ‘antedrug concept’
to reduce the systemic side effects [14]. A synthesized
FP-ISO-21AC showed higher ability to inhibit the cyto-
kine-induced production of NO than PRED. There was
no significant difference between FP-ISO-21AC and
PRED on inhibitory effect on cytokine-induced IL-8
expression, but the FP-ISO-21AC showed higher inhibi-
tory effect on cytokine-induced NO production (Figure
2) and also enhanced IL-22R expression (Figure 4A)
with less systemic side effects.
3.1. Anti-Inflammatory Steroidal Antedrug,
FP-ISO-21AC, and Its Parent PRED
Inhibit Cytokine-Induced NO Production
Figure 2 shows the inhibitory effects of anti-inflamma-
tory steroidal antedrugs, FP-ISO-21AC and its parent
prednisolone (PRED), on cytokine-induced NO produc-
tion. Both FP-ISO-21AC and PRED inhibited cyto-
kine-induced NO production. One hour pre-treatment of
FP-ISO-21AC at 1 M reduced cytokine-induced NO
production down to 37%. PRED at 1 M reduced cyto
kine-induced NO production down to 65%. The FP-ISO-
21AC showed higher inhibitory effect on cytokine-ind-
uced NO production. In other words, the FP-ISO-21AC
showed higher anti-inflammatory effect.
3.2. Anti-Inflammatory Steroidal Antedrug
FP-ISO-21AC and Its Parent PRED
Inhibit Cytokine-Induced Interleukin-8
(IL-8) Production
Anti-inflammatory steroidal antedrug FP-ISO-21AC and
0
20
40
60
80
100
120
% inhibition of NO production
cytomix without steroids
cytomix with 1μM PRED
pretreatment
cytomix with 1μM FP-
ISO-AC pretreatment
Figure 2. Inhibitory effect of steroids on cytokine-induced NO
production in A549 human alveolar epithelial cells. Cells were
exposed cytomix (The mixture of the three cytokines IL-1,
TNF- and IFN- and the concentration of each cytokine is 50
ng/ml) following pretreatment of steroids at indicated concen-
trations for 1 hour. The concentration of NO was measured 24
hours after cytokine exposure by amiNO series of NO sensors.
Data from steroids treatment experiments were expressed as
percentage of untreated controls. p≤0.05.
0
20
40
60
80
100
120
% inhibition of IL-8 production
cytomix without steroids
cytomix with 1µM PRED
pretreatment
cytomix with 1µM FP-ISO-
21AC pretreatment
Figure 3. Inhibitory effect of steroids on cytokine-induced
IL-8 production in A549 human alveolar epithelial cells. Cells
were exposed cytomix (IL-1β, TNF-α and IFN-γ at 50 ng/ml
respectively) following pretreatment of steroids at indicated
concentrations for 1 hour. The concentration of IL-8 was
measured by ELISA after 24 hours exposure to cytokine. Data
from steroids treatment experiments were expressed as per-
centage of untreated controls. p≤0.05. One hour pretreatment
of FP-ISO-21AC and PRED at 1 M reduce cytokine-induced
IL-8 production to 50% and 53% respectively.

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130
PRED inhibited cytokine-induced IL-8 production (Fig-
ure 3). The data showed that one hour pretreatment of
FP-ISO-21AC at 1 µM reduced cytokine-induced IL-8
production down to 50%. PRED reduced cytokine - in-
duced IL-8 production down to 53%. There was no sig-
nificant difference between FP-ISO-21AC and PRED on
the inhibitory effect on cytokine-induced IL-8 expres-
sion.
3.3. Effects of Anti-Inflammatory Steroidal
Antedrug on Inhibitory Cytokine
As shown in Figure 4, PRED and FP-ISO-21AC sig-
nificantly reduced the cytokine-induced NO production.
One hour pre-treatment of PRED and FP-ISO-21AC at 1
µM reduced cytokine-induced NO production down to
57% and 43%, respectively. In the presence of exoge-
nous IL-22, a further reduction about 17% (39% and
26%, respectively for FP-ISO-21AC and PRED) of cy-
tokine-induced NO production in A549 cells pre-treated
with anti-inflammatory steroids (Figure 4A) was ob-
served. Interestingly, there was no significant effect of
exogenous IL-22 on anti-inflammatory steroids in cyto-
kine-induced NO production from HLF1 (Figure 4B).
4. DISCUSSION
Asthma is a chronic, episodic disease of the airways
characterized by repeated exacerbations of bronchial
airway narrowing and difficult breathing. Chronic and
persistent bronchial asthma is life-threatening and dis-
tresses more than 5% of the population in the United
States alone and the incidence has been increasing since
the 1980’s [20,21]. In asthma treatment, consistent ef-
forts have been made in searching for effective steroids
with least side effects. In this direction, a considerable
amount of research has been conducted to increase the
therapeutic index of potent corticosteroids and reducing
their systemic side effects. Notably, FP-ISO-21AC was
designed with antedrug concept and was shown to have
less systemic side effects [10]. It has higher anti-in-
flammatory activities than parent steroid PRED [14,17,
18]. Corticosteroids are recently shown to inhibit the
expression of pro-inflammatory mediators such as NO
[22], IL-8 [23,24] and IL-5 [24], with their stimulated
release of the anti-inflammatory cytokine such as IL-10
[25].
However, it is unclear whether structural modification
of corticosteroids may affect these inhibitory pathways
at sites of inflammation. The present study is an attempt
to better understand the antedrug switch off multiple
pro-inflammatory pathways and switch on anti-inflam-
matory pathways in asthma with further investigations
on NO production. In this study, anti-inflammatory ster-
oidal antedrug FP-ISO-21AC, which was derived from
PRED was used to test its effects on inhibiting NO pro-
(A) A549 cells
0
20
40
60
80
100
120
ControlCytomix Cytomix+IL-22
% Decrease in NO production
No ante-drugPRED PretreatedFP-ISO-21AC Pretreated
(B) HLF1 cells
0
20
40
60
80
100
120
ControlCytomix Cytomix+IL-22
% Decrease in NO production
no ante-drugPRED pretreatedFP-ISO-21AC pretreated
Figure 4. (A) Effects of anti-inflammatory steroidal antedrug
FP-ISO-21AC and PRED on inhibitory cytokine in cyto-
kine-induced NO production in A549 cells. FP-ISO-21AC
and PRED further reduced cytokine induced NO production in
the presence of exogenous IL-22. (B) Effects of anti-inflam-
matory steroidal antedrug FP-ISO-21AC and PRED on inhibi-
tory cytokines in cytokine-induced NO production in HLF1.
No further reduced cytokine induced NO production was ob-
served in the presence of exogenous IL-22. Cells were stimu-
lated with three cytokines (IL-1, TNF-, and IFN-, 50
ng/mL respectively) or three cytokines (IL-1, TNF-, and
IFN-, 50 ng/mL respectively) and IL-22 (20 ng/mL) in com-
bination. After 24 hours incubation, nitric oxide was measured.
Data from steroids treatment experiment were expressed as
percentage of untreated controls, p≤0.05.
duction in human alveolar epithelial cells (A549). Our
results show that both FP-ISO-21AC and PRED inhib-
ited the cytokine-induced NO production. One hour pre-
treatment of FP-ISO-21-AC at 1 M reduced cytokine-
induced NO production down to 37%. The PRED
showed lower inhibitory effect than FP-ISO-21AC and

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reduced cytokine-induced NO production down to 65%
(Figure 2). There was no significant difference between
FP-ISO-21AC and PRED on inhibitory effect on cyto-
kine-induced IL-8 expression, but the FP-ISO-21AC
showed higher inhibitory effect on cytokine-induced NO
production (Figure 2).
To confirm the observed decrease in NO production
related to enhanced expression of inhibitory cytokines,
exogenous IL-22 was added to the media of A549 in the
presence or absence of anti-inflammatory steroids. IL-22
is a member of the human type I IFN family, which in-
cludes IL-10. Our data corroborates with earlier report
on IL-22 effect in the presence of inflammatory steroids
[26]. Other studies support our observations on lung
epithelial cells (A549) which express IL-22R1 (inter-
leukin 22 receptor) subunit in response to IL-22 [15]. As
the results indicated in Figure 4A and 4B, both PRED
and FP-ISO-21AC may enhance the expression of
IL-22R1, and increase the responsiveness to exogenous
IL-22 in A549 cells which do express IL-22R1.
An important factor is the balance between the effects
of pro-inflammatory and anti-inflammatory chemokines
or cytokines in determining the severity of the inflam-
matory disease. Corticosteroids have effects on expres-
sion of cytokines receptors [27,28]. Corticosteroids are
understood to upregulate cytokine receptor expression
that correlates with enhanced cytokine effects on target
cells [27]. Other earlier studies also suggest that recep-
tors for IL-1 [29,30], IL-2 [31], IL-4 [32], IL-6 [33,34],
IFN- [35] and GM-CSF [36] are all up-regulated by
glucocorticoids.
Inflammatory stimuli increase the expression of many
inflammatory genes, resulting in the inflammatory re-
sponses. The same stimuli may also induce inhibitory
cytokines, which can inhibit the expression of these in-
flammatory proteins, thus terminating this inflammatory
response. There has been increasing evidence that a de-
fective expression of inhibitory cytokines is observed in
patients with asthma [37]. The reduced expression of
inhibitory cytokines leads to increased and more pro-
longed inflammation. Subsequent treatment by anti-in-
flammatory steroids can restore the secretion of inhibi-
tory cytokines [37]. Structural modification of corticos-
teroids may restore this defective expression of inhibi-
tory cytokines. The structural modification can be an
important direction forwards the design of novel anti-in-
flammatory steroidal antedrugs for the treatment of
asthma.
There has been increasing evidences that the role of
inhibitory cytokines is crucial on the treatment of asthma
[38]. IL-22 is one of the most important inhibitory cyto-
kines involved in the inhibition of the inflammation. It
can regulate the expression of proinflammatory cyto-
kines [39]. However, the effect of steroid on IL-22R is
not known. In order to test if steroids can enhance the
expression of IL-22R, the effects of IL-22 on the inhibi-
tion of NO production induced by the pre-treatment of
anti-inflammatory steroids was tested. Our data demon-
strated that in the presence of the exogenous inhibitory
cytokine IL-22, further reduction of NO production was
observed in A549 cells (Figure 4A) as earlier demon-
strated to express IL-22R [39] but not in HLF1 (Figure
4B) which do not express IL-22R. These data suggested
that the steroidal ante-drugs may enhance the expression
the IL-22 receptor. FP-ISO-21AC showed higher po-
tency than PRED to restore the expression of IL-22R.
FP-ISO-21AC further reduced NO production to 27%
and PRED further reduced NO production to 39%.
In conclusion, a synthesized antedrug FP-ISO-21AC
showed higher anti-inflammatory effects than PRED by
inhibiting the expression of pro-inflammatory mediator
NO. There was no significant difference between
FP-ISO-21AC and PRED on inhibitory effect on cyto-
kine-induced IL-8 expression, but the FP-ISO-21AC
showed higher inhibitory effect on cytokine-induced NO
production. FP-ISO-21AC also showed higher potency
than its parent PRED in stimulating the possible expres-
sion of IL-22R. This study showed that structural modi-
fication of anti-inflammatory steroids with ‘antedrug
concept’ (with the direction of reducing the systemic
side effects) could enhance their anti-inflammatory ac-
tivities with less systemic side effects.
REFERENCES
[1] D. T. Boumpas, G. P. Chrousos, R. L. Wilder, T. R. Cupps,
J. E. Balow, (1993) Glucocorticoid therapy for immune-
mediated diseases - basic and clinical correlates, Annals
of Internal Medicine, 119, 1198-1208.
[2] J. W. Funder, (1997) Glucocorticoid and mineralocorti-
coid receptors: biology and clinical relevance, Annual
Review of Medicine, 48, 231-240.
[3] H. M. Reichardt, G. Schutz, (1998) Glucocorticoid sig-
nalling - multiple variations of a common theme, Mo-
lecular and Cellular Endocrinology, 146, 1-6.
[4] L.I. McKay, JA. Cidlowski, (1999) Molecular control of
immune/inflammatory responses: Interactions between
nuclear factor-kappa B and steroid receptor-signaling
pathways, Endocrine Reviews, 20, 435-459.
[5] L. A. Wolfraim, (2006) Treating autoimmune diseases
through restoration of antigen-specific immune tolerance,
Archivum Immunologiae Et Therapiae Experimentalis,
54, 1-13.
[6] H. Schacke, W. D. Docke, K. Asadullah, (2002) Mecha-
nisms involved in the side effects of glucocorticoids,
Pharmacology & Therapeutics, 96, 23-43.
[7] E. Garbe, J. LeLorier, J. F. Boivin, S. Suissa, (1997) Risk
of ocular hypertension or open-angle glaucoma in elderly
patients on oral glucocorticoids, Lancet, 350, 979-982.
[8] P. B. Jacobson, T. W. von Geldern, L. Ohman, M. Oster-
land, J. H. Wang, B. Zinker, D. Wilcox, PT. Nguyen, A.
Mika, S. Fung, T. Fey, A. Goos-Nilsson, M. Grynfarb, T.

G. F. Wang et al. / HEALTH 1 (2009) 127-133
SciRes Copyright © 2009 Openly accessible at http://www.scirp.org/journal/HEALTH/
132
Barkhem, K. Marsh, D. W. A. Beno, B. Nga-Nguyen, P. R.
Kym, J. T. Link, N. Tu, DS. Edgerton, A. Cherrington,
S. Efendic, B. C. Lane, T. J. Opgenorth, (2005) Hepatic
glucocorticoid receptor antagonism is sufficient to reduce
elevated hepatic glucose output and improve glucose
control in animal models of type 2 diabetes, Journal of
Pharmacology and Experimental Therapeutics, 314, 191-
200.
[9] J. Nishimura, S. Ikuyama, (2000) Glucocorticoid-Induced
osteoporosis: pathogenesis and management, Journal of
Bone and Mineral Metabolism, 18, 350-352.
[10] H. J. Lee, MRI. Soliman, (1982) Anti-Inflammatory ster-
oids without pituitary-adrenal suppression, Science, 215,
989-991.
[11] H. J. Lee, M. A. Khalil, J. W. Lee, (1984) Antedrug - a
conceptual basis for safer anti-inflammatory steroids,
Drugs under Experimental and Clinical Research, 10,
835-844.
[12] H. J. Lee, R. Trottier, (1980) Antiinflammatory activity
of two novel derivatives of prednisolone, Res Commun
Chem Pathol Pharmacol, 27, 611-4.
[13] H. Ueno, A. Maruyama, M. Miyake, E. Nakao, K. Nakao,
K. Umezu, I. Nitta, (1991) Synthesis and evaluation of
antiinflammatory activities of a series of corticosteroid
17-Alpha-Esters containing a Functional-Group, Journal
of Medicinal Chemistry, 34, 2468-2473.
[14] K. K. Park, D. H. Ko, Z. You, M. O. F. Khan, H. J. Lee,
(2006) In vitro anti-inflammatory activities of new ster-
oidal antedrugs: [16 alpha, 17 alpha-d] isoxazoline and
[16 alpha, 17 alpha-d]-3'-hydroxy-iminoformyl isoxazo-
line derivatives of prednisolone and 9 alpha - fluoropred-
nisolone, Steroids, 71, 183-188.
[15] H. A. Whittington, L. Armstrong, K. M. Uppington, A.B.
Millar, (2004Interleukin-22 - A potential immunomodu-
latory molecule in the lung, American Journal of Respi-
ratory Cell and Molecular Biology, 31, 220-226.
[16] B. M. Necela, J. A.Cidlowski, (2003) Crystallization of
the human glucocorticoid receptor ligand binding domain:
a step towards selective glucocorticoids, Trends in Phar-
macological Sciences, 24, 58-61.
[17] M. A. Khalil, M. F. Maponya, D. H. Ko, Z. Q. You, E. T.
Oriaku, H. J. Lee, (1996) New anti-inflammatory steroids:
[16 alpha, 17 alpha-d] isoxazoline derivatives of predni-
solone and 9 alpha-fluoroprednisolone, Medicinal Chem-
istry Research, 6, 52-60.
[18] D. H. Ko, M. F. Maponya, M. A. Khalil, E. T. Oriaku,
Z. Q. You, H. J. Lee, (1997) New anti-inflammatory ster-
oids: [16 alpha, 17 alpha-d] -3'-hydroxyiminoformyl
isoxazoline derivatives of prednisolone and 9 apha- flu ro-
prednisolone. Medicinal Chemistry Research, 7, 313-323.
[19] T. Kwon, A. S. Heiman, E. T. Oriaku, K. Yoon, H. J. Lee
(1995) New steroidal antiinflammatory antedrugs - ster-
oidal [16- Alpha,17-Alpha-D]-3'-Carbethoxyisoxazolines.
Journal of Medicinal Chemistry, 38, 1048-1051.
[20] JA. Elias, Z. Zhu, G. Chupp, RJ. Homer, (1999) Airway
remodeling in asthma, Journal of Clinical Investigation;
104, 1001-1006.
[21] J. Elias, (1999) Inspirations on asthma, Journal of Clini-
cal Investigation, 104, 827-827.
[22] N. K. Worrall, T. P. Misko, P. M. Sullivan, J. J. Hui, C. P.
Rodi, T. B. Ferguson, (1996) Corticosteroids inhibit ex-
pression of inducible nitric oxide synthase during acute
cardiac allograft rejection, Transplantation, 61, 324-328.
[23] K. Nyhlen, M. Linden, R. Andersson, S. Uppugunduri,
(2000) Corticosteroids and interferons inhibit cyto-
kine-induced production of IL-8 by human endothelial
cells, Cytokine, 12, 355-360.
[24] B. Wallwork, W. Coman, F. Feron, A. Mackay-Sim, A.
Cervin, (2002) Clarithromycin and prednisolone inhibit
cytokine production in chronic rhinosinusitis, Laryngo-
scope, 112, 1827-1830.
[25] M. John, S. Lim, J. Seybold, P. Jose, A. Robichaud, B.
O'Connor, P. J. Barnes, K. F. Chung, (1998) Inhaled cor-
ticosteroids increase interleukin-10 but reduce macrophage
inflammatory protein-1 alpha, granulocyte-macrophage
colony-stimulating factor, and interferon-gamma release
from alveolar macrophages in asthma, American Journal
of Respiratory and Critical Care Medicine, 157, 256-262.
[26] L. Dumoutier, J. Louahed, J. C. Renauld, Cloning and
characterization of IL-10-related T cell-derived inducible
factor (IL-TIF): a novel cytokine structurally related to
IL-10 and inducible by IL-9, Journal of Immunology
l2000, 164, 1814-1819.
[27] W. Y. Almawi, H. N. Beyhum, A. A. Rahme, M. J. Rieder,
(1996) Regulation of cytokine and cytokine receptor ex-
pression by glucocorticoids, Journal of Leukocyte Biol-
ogy,60, 563-572.
[28] G. J. Wiegers, J. M. H. M. Reul, (1998) Induction of
cytokine receptors by glucocorticoids: functional and
pathological significance, Trends in Pharmacological
Sciences, 19, 317-321.
[29] T. Akahoshi, J. J. Oppenheim, K. Matsushima, (1988)
Induction of high-affinity Interleukin-1 receptor on hu-
man peripheral-blood lymphocytes by glucocorticoid
hormones, Journal of Experimental Medicine, 167, 924-
936.
[30] P. E. Gottschall, K. Koves, K. Mizuno, I. Tatsuno, A.
Arimura, (1991) Glucocorticoid up-regulation of Inter-
leukin-1 receptor expression in a glioblastoma Cell-Line,.
American Journal of Physiology, 261, E362-E368.
[31] G. J. Wiegers, M. S. Labeur, I. E. M. Stec, W. E. F.
Klinkert, F. Holsboer, J. M. H. M. Reul, (1995) Gluco-
corticoids accelerate anti-T cell receptor-induced T-cell
growth, Journal of Immunology, 155, 1893-1902.
[32] R. L. K. Paterson, R. Or, J. M. Domenico, G. Delespesse,
E. W. Gelfand, (1994) Regulation of Cd23 expression by
Il-4 and corticosteroid in human B-lymphocytes - altered
response after Ebv infection, Journal of Immunology,
152, 2139-2147.
[33] L. Snyers, L. Dewit, J. Content, (1990) Glucocorticoid
up-regulation of high-affinity Interleukin-6 receptors on
human Epithelial-Cells, Proceedings of the National
Academy of Sciences of the United States of America, 87,
2838-2842.
[34] S. P. Campos, Y. P. Wang, A. Koj, H. Baumann, (1993)
Divergent transforming Growth-Factor-Beta effects on
Il-6 regulation of Acute-Phase Plasma-Proteins in Rat
Hepatoma-Cells, Journal of Immunology, 151, 7128-7137.
[35] R. W. Strickland, L. M. Wahl, D. S. Finbloom, (1986)
Corticosteroids enhance the binding of recombinant in-
terferon-gamma to cultured human-monocytes, Journal
of Immunology, 137, 1577-1580.
[36] C. M. Hawrylowicz, L. Guida, E. Paleolog, (1994) Dex-
amethasone up-regulates granulocyte-macrophage colony

G. F. Wang et al. / HEALTH 1 (2009) 127-133
SciRes Copyright © 2009 http://www.scirp.org/journal/HEALTH/Openly accessible at
133
133
- stimulating ractor-receptor expression on human mono-
cytes, Immunology, 83, 274-280.
[37] P. J. Barnes, S. Lim, (1998) Inhibitory cytokines in
asthma, Molecular Medicine Today, 4, 452-458.
[38] D. Kunz, G. Walker, W. Eberhardt, J. Pfeilschifter, (1996)
Molecular mechanisms of dexamethasone inhibition of
nitric oxide synthase expression in interleukin 1 beta-
stimulated mesangial cells: evidence for the involvement
of transcriptional and posttranscriptional regulation,
Proceedings of the National Academy of Sciences of the
United States of America, 93, 255-259.
[39] D. A. Geller, A. K. Nussler, M. Disilvio, C. J. Lowenstein,
R. A. Shapiro, S. C. Wang, R. L. Simmons, T. R. Billiar,
(1993) Cytokines, endotoxin, and glucocorticoids regu-
late the expression of inducible Nitric-Oxide synthase in
hepatocytes, Proceedings of the National Academy of
Sciences of the United States of America, 90, 522-526.