Lem.
`
` ida-
`
`sntiallergika -Antiasthmatika - Antitussiva - Bronchodilatatoren -
`Bronchosekretolytika - Mukolytika
`‘
`
` ./\l"/HP) m Forsch Drug
`
`hi Antiallergic Drugs - Antiasthmatics - Antitussives - Bronchodilators -
`' Bronchosecretogogues - Mucolytics
`
`Effects of Antihistamines on
`
`Leukotriene and Cytokine Release
`from Dispersed Nasal Polyp Cells
`
`9
`
`Sabine Kiisters “, Rufina Schuligoi 1’, Karl-Bernd Hiittenbrink '3, Iutta Rudert “, Angela Wachs 3,
`Istvan Szelenyi 3, and Bernhard A. Peskarb
`
`Pulmonary Pharmacology, Corporate Research ASTA Medica AG / Arzneimittelwerk Dresden GmbHa,
`Radebeul>(Germany), Institute of Experimental and Clinical Pharmacology, University of Graz b, Graz (Austria),
`and Clinic for Otolaryngology, Dresden University of Technologyc, Dresden (Germany)
`
`Key words
`
`I Azelastine
`I Cetirizine
`
`I leukotriene
`I_
`
`
`
`Summary
`
`In this study the effects of antihistamines
`on the release of eicosanoids and the
`
`pro-inflammatory cytokine tumor necro-
`sis factor alpha (TNFa) were compared.
`Enzymatically dispersed cells from hu-
`man nasal polyps served as an in vitro
`model of chronic respiratory mucosal in-
`flammation. Nasal polyp cells (2 x 105/
`ml) were sensitized with human IgE pre-
`incubatediwith azelastine (CAS 58581-89-
`8), terfenadine (CAS 50679-08-8), levocab-
`astine (CAS 79516-68-0) or cetirizine
`(CAS 83881-51-0), and stimulated with
`anti—human immunoglobulin E (IgE).
`Thromboxane B2 (TBX2) and leukotriene
`C4 (LTC4) were measured by radio-
`immunoassay (RIA), TNFa by enzyme-lin-
`ked immunosorbent assay (ELISA). Data
`represent mean values of % inhibition
`estimated from the untreated positive
`
`control or mean IC50 (n = 5). Azelastine
`and terfenadine inhibited TNFa release
`with IC50 values of 6.2 pmol/1 and 4.3
`umol/l, respectively. Terfenadine reduced
`TXB2 release by 37 i 15 %, and LTC4 re-
`lease was decreased by azelastine and ter-
`fenadine very potently by 86 % and
`100 %, respectively. Azelastine shows
`anti—inflammatory properties in thera-
`peutically relevant concentrations as as-
`sessed by its ability to reduce TNFu re-
`lease as well as its ability to inhibit LTC4
`production in allergically stimulated hu-
`man nasal polyp cells.
`
`
`
` Zusammenfassung
`
`Wirkimg von Antihistaminika auf die
`Freisetzung von Leukotrien und Cytokin
`aus dispergierten Zellen nasaler Polypen
`
`In der vorliegenden Studie wurde die
`Wirkung von Antihistaminika auf die
`Freisetzung von Eicosanoiden und des
`
`entziindungsfiirdernden Cytokins ,,Tu-
`mor necrosis factor“ a (TNFa) unter-
`sucht. Enzymatisch dispergierte Zellen
`von menschlichen nasalen Polypen dien-
`ten als In-vitro-Modell der chronischen
`
`mukosalen Entziindung der Atemwege.
`Diese Zellen 2 x 10"/ml) wurden mit
`
`
`
`Exhibit 1016
`Exhibit 1016
`IPR2017-00807
`|PFl2017-00807
`Kiisters et al. — Antihistamines
`ARGENTUM
`ARGENTUM
`
`
`Armeim.-Forsch.lI)rug Res. 52, No. 2, 97-102 (2002)
`© ECV - Editio Cantor Verlag, Aulendorf (Germany)
`
`000001
`000001
`
`

`

`
`
`
`
`
`menschlichem Immunglobulin E (IgE)
`sensibilisiert. Anschlieliend wurden sie
`
`mit Azelastin (CAS 58581-89-8), Terfena-
`din (CAS 50679-08-8), Levocabastin (CAS
`79516-68-0) oder Cetirizin (CAS 83881-
`51-0) vorinkubiert. Die allergische Stimu-
`lation erfolgte mit Anti-human-IgE.
`Thromboxan B2 (TXB2) und Leukotrien
`C4 (LTC4) wurden mit Hilfe von RIA (ra-
`dioimmunoassay), TNFa mit ELISA (en-
`
`Antiallergika - Antiasthmatika - Antitussiva - Bronchodilatatoren ~ Bronchosekretolytika « Mukolytika
`
`zyme-linked immunosorbent assay) ge-
`messen. Die Ergebnisse entsprechen ei-
`ner prozentualen Hemmung im Vergleich
`zu nicht-behandelten Positivkontrollwer-
`ten (11 = 5). Azelastin und Terfenadin
`hemmten die TNF¢1-Freisetzung mit IC50-
`Werten von 6,2 pmol/I bzw. 4,3 pmol/l.
`Terfenadin reduzierte die TXB2-Freiset-
`zung um 37 + 15 %. Die LTC4-Freisetzung
`wurde durch Terfenadin und Azelastin
`
`
`sehr stark, um 86 % bzw. 100 % ge-
`hemmt. In therapeutisch relevanten Kon-
`zentrationen besitzt Azelastin entziin-
`dungshemmende Eigenschaften, die
`durch die Hemmung der TNFa- bzw. der
`LTC4-Freisetzung charakterisiert werden
`konnten.
`
`1 . Introduction
`
`Histamine H1-receptor antagonists together with top-
`ical steroids are the treatment of choice in allergic rhin-
`itis. The phthalazinone derivative azelastine has suc-
`cessfully been introduced in the therapy of allergic rhin-
`itis and conjunctivitis. It has also been demonstrated
`that azelastine reduces nasal congestion [I-3]. This fea-
`ture, which disginguishes azelastine from oral antihis-
`tamines, is of great interest, because corticosteroids are
`known to be quite effective in relieving nasal conges-
`tion, whereas antihistamines are effective for the sneez-
`
`ing, itching, watery secretion, but not congestion.
`It is well known that azelastine and some other anti-
`
`histamines may possess properties unrelated to block-
`ade of histamine at its receptor level. In animal studies,
`it has been demonstrated that in addition to acting as
`a histamine Hyreceptor antagonist, azelastine also in-
`hibits the production of many chemical mediators of
`the allergic response [4—9]. Azelastine has been re-
`ported to inhibit leukotriene (LT)-mediated acute aller-
`gic bronchoconstriction in guinea pigs and passive cu-
`taneous anaphylaxis in rats [10, 11]. In an in vitro study
`using chopped lungs of actively sensitized guinea pigs,
`we have shown that azelastine is able to inhibit the al-
`
`lergically induced release of cysteinyl—LTs [12].
`There is evidence that azelastine is able to inhibit the
`
`release of cysteinyl-LTs from different cell types, includ-
`ing human cells [4, 13-16]. Moreover, polypous tissue
`samples have been shown to produce more cysteinyl-
`LT upon stimulation than normal nasal mucosa indicat-
`ing an altered pattern of arachidonic acid pathways
`[17]. Therefore, the present study has been performed
`with enzymatically dispersed cells obtained from hu-
`man nasal polyps to compare the effect of antihistam-
`ines on the release of pro—inflammatory cytoldnes and
`eicosanoids. Nasal polyps are a model of chronic respir-
`atory mucosal inflammation [18]. Not only residing
`cells such as epithelial cells, monocytes, macrophages,
`and mast cells are contained in this cell culture; also
`
`infiltrated cells such as immunocompetent lympho-
`cytes, neutrophil granulocytes and often a high amount
`of eosinophils can be found. Hence, this culture system
`ensures an intensive interplay between the different cell
`populations,
`thereby providing a suitable model
`to
`
`study the pharmacologic action of anti-inflammatory
`and anti-asthmatic drugs [l9—22].
`
`2. Materials and methods
`
`2.1. Human nasal polyp cells
`
`Nasal polyps were obtained from patients with nasal polyposis
`after polypectomy (n : 48). Patients had been free of any med-
`ication for at least 2 weeks. The study was approved by the
`local ethics committee. The polyps were washed in RPMI 1640
`(Biochrom, Berlin, Germany), dissected and cell disaggregation
`was achieved by enzymatic digestion according to Campbell
`and Bousquet [23]. Briefly, the minced polyps were incubated
`for 2 h at 37 °C in RPMI 1640 (I g tissue per 4 ml) containing
`2.0 mg/ml protease, 1.5 mg/ml collagenase, 0.75 mg/ml hyalu-
`ronidase and 0.05 mg/ml DNase (all enzymes Sigma, Deisen-
`hofen, Germany). Then, the cell suspension was filtered to re-
`move any undigested tissue and the cells were washed 3 times.
`After an incubation in the presence of human IgE (1 ug/ ml in
`RPMI 1640 10 % fetal calf serum (FCS), Biochrom) for I h at
`37 °C, the cells were washed two times, Counted and resuspen-
`ded at a concentration of 2 X 106 cells/ml. Then 1 ug anti—lgE
`antibodies (Calbiochem, Bad Soden, Germany) per well were
`added to the cell suspension (1 ml per well in 6 well plates) to
`mimic an allergen challenge by cross-linking the receptor
`bound IgE antibodies. After enzymatic disaggregation, cells
`were counted in a Neubauer chamber using trypan blue stain-
`ing. Cell viability always exceeded 95 %.
`
`2.2. Compounds
`Azelastine hydrochloride (CAS 79307-93-0) was synthesised by
`the Chemical Department of ASTA Medica AG (Frankfurt, Ger-
`many). Terfenadine (CAS 50679-08)
`levocabastine hydro-
`chloride (CAS 79547—78—7) and cetirizine hydrochloride (CAS
`83881-52-1) were kindly provided by the respective manufac-
`turers.
`
`2.3. Qrtokine determination
`To determine the effect of antihistamines on cytokine release
`nasal polyp cells were pre—incubated with 3, 10 and 30 umol/l
`of either azelastine, terfenadine, cetirizine and levocabastine
`respectively or with vehicle (0.1 % DMSO) for 30 min prior to
`addition of anti—IgE. After an incubation period of 18 h (37 °C
`and 5 % CO2) the cell suspension was centrifuged and the su-
`pernatant was stored at -70 °C until determination of TNFa
`contents by sandwich ELISA using an antibody pair and stand-
`ard protein from PharMingen (Hamburg, Germany).
`
`98 Kiisters et al. — Antihistamines
`
`000002
`000002
`
`Arzneim.-Forsch.IDrug Res. 52, No. 2, 97-102 (2002)
`© ECV ~ Editio Cantor Verlag, Aulendorf (Germany)
`
`
`
`.......,.,..-_-.-,..~./~...-._.p,,.;--.
`
`
`
`

`

`
`
`
`Antia||ergic‘Drugs - Antiasthmatics - Antitussives ~ Bronchodilators - Bronchosecretogogues - Mucolytlcs
`
`—O— terfenadine
`°—V— levocabastine
`—V— cetirizine
`
`cno
`
`60
`
`
`
`inhibitionofTNFcLrelease[%ofcontrol]
`
` -0- azelastine
`
`Fig. 1: Inhibition of allergically induced release of TNF(‘l from e
`zymatically dispersed nasal polyp cells. Nasal polyp cells (2 X ll
`ml) were sensitized with human IgE for 1 h. 30 min prior
`stimulation with anti-human IgE 3, 10, or 30 umol/1 azelastir
`terfenadine, levocabastine and cetirizine were added respective
`18 h after anti-IgE addition supernatants were removed a]
`frozen at -70 °C until cytokine determination with ELISA. Dz
`represent mean values of % inhibition estimated from the u
`treated positive control i SD (n = 5)
`
`and LTC4 in parallel. Positive control nasal polyp cel
`which were only sensitized with human IgE and stim
`lated with anti—human lgE, released 320 pg/ml TX
`and 350 pg/ ml LTC4. Azelastine, terfenadine, levocaba
`tine or cetirizine were given to the nasal polyp cells
`a concentration of 10 pmol/l 30 min before addition
`the allergic stimulus anti—lgE. Of all the antihistamin
`investigated only terfenadine reduced TXB2 release l
`37 i 15 % as compared to the positive control assess:
`one hour after addition of anti-IgE. On average neith
`
`
`
`
`100
`
`80
`
`60
`
`40
`
`20
`
`
`
`“/ninhibitionofLTC4release
`
`azelastine
`
`terfenadine
`
`levocabastine
`
`Fig. 2: Inhibition of allergically induced release of LTC4 from e
`zymatically dispersed nasal polyp cells. Nasal polyp cells (2 x ll
`ml) were sensitized with human IgE for 1 h. Thirty minutes pri
`to stimulation with anti—human IgE 10 pmolll azelastine, terfen
`dine and levocabastine were added respectively. One hour afl
`anti-IgE addition supernatants were removed, shock frozen in
`quid nitrogen and stored at -70 °C until LTC4 determinatic
`Data represent mean values of % Inhibition estimated from t
`untreated positive control i SD (n = 5).
`
`2.4. Eicosanoid determination
`To determine the effect of antihistamines on eicosanoid release
`
`nasal polyp cells were pre-incubated with 10 umol/l of either
`azelastine,
`terfenadine, cetirizine and levocabastine respec-
`tively or with vehicle (0.1 % DMSO) for 30 min prior to addition
`of anti-IgE. After an incubation period of 1 h (37 °C and 5 %
`CO2) the suspension was centrifuged; the supernatant was im-
`mediately frozen in liquid nitrogen and stored at ~70 °C until
`determination of eicosanoid contents.
`
`Immunoreactive TXB2 and cysteinyl-LT (calculated using a
`LTC4 standard curve and expressed as LTC4 equivalents [24])
`were determined as described previously [25, 26], using [5, 6,
`8, 9, ll, 12, 14, 15(n)3H]TXB2 and [14, 15, 19, 20-3H(n)]LTC4
`(both from New England Nuclear, Vienna, Austria) as radio-
`ligands and synthetic TXB2 and LTC4 (both from Cayman, Ann
`Arbot, USA) as standards, respectively.
`
`2.5. Analysis of results
`For determination of the percent inhibition of mediator release
`by antihistamines the results were compared to the positive
`control and the average inhibition for each concentration was
`calculated. Data are given in mean i SD. The lC50 values were
`determined using an in-house programmed PC software pack-
`age (EDX 2.1).
`
`3. Results
`
`3.1. Effect of antihistamines on TNFu release
`from nasal polyp cells
`
`The finding that azelastine may act via mechanisms dif-
`ferent from blocking the histamine receptor prompted
`us to investigate its anti-inflammatory potency. There-
`fore the ability of azelastine to reduce the allergically
`triggered release of the pro—inflammatory cytokine
`TNFa was tested and compared to that of terfenadine,
`levocabastine and cetirizine. Nasal polyp cells, which
`were only sensitized with human IgE and stimulated
`with anti—human 1gE, served as positive control and re-
`leased 400 to 500 pg/ ml TNFCL into the supernatant 18 h
`after addition of the stimulus. Azelastine and terfenad-
`ine inhibited the release of TNFa concentration-de-
`
`pendently and were very potent in this respect, since
`they almost completely prevented TNFOL release at 30
`umol/l (Fig. 1). Both antihistamines were equally effec-
`tive with an IC50 Value of 6.2 umol/l for terfenadine and
`4.3 umol/l for azelastine. At a concentration of 3 pmol/l
`levocabastine and cetirizine were as effective as terfen-
`
`adine in inhibiting TNFo release by 40 % and 35 %, re-
`spectively. However, an increase in the concentration of
`either antihistamine hardly enhanced their inhibitory
`action (Fig. 1).
`
`3.2. Effect of antihistamines on TXB2 and LTC4
`release from nasal polyp cells
`
`To further investigate possible actions of antihistamines
`which are not mediated through blockade of histamine
`receptors we determined the influence on the produc-
`tion of arachidonic acid metabolites. To investigate
`whether the cyclic or the linear pathway of arachidonic
`acid metabolism was influenced, we determined TXB2
`
`zvanten Kon-
`1 entziin—
`ten, die
`341- bzw. der
`iert werden
`
` ~ % ge-
`
`1| lll !
`
`
`
`_.__....._._.....__._.._...:_...__.—-_.___.._..._
`
`Elammatory
`
`tsal polyposis
`2 of any med-
`roved by the
`in RPMI 1640
`
`.isaggregation
`to Campbell
`are incubated
`11) containing
`mg/ ml hyalu—
`gma, Deisen-
`filtered to re-
`rshed 3 times.
`
`;E (1 ug/ ml in
`In) for 1 h at
`and resuspen-
`1 pg anti—IgE
`per well were
`well plates) to
`the receptor
`regation, cells
`an blue stain-
`
`;ynthesised by
`Frankfurt, Ger-
`astine hydro-
`chloride (CAS
`tive manufac-
`
`rtokine release
`
`and 30 umol/l
`levocabastine
`
`0 min prior to
`of 18 h (37 °C
`ed and the su-
`ation of TNFa
`)air and stand-
`Lny).
`
`. 2, 97-102 (2002)
`lendorf (Germany)
`
`Armeim.-Forsch./Drug Res. 52, No. 2, 97-102 (2002)
`© ECV - Editio Cantor Verlag, Aulendorf (Germany)
`
`000003
`000003
`
`Kiisters et al. — Antihistamines
`
`9
`
`

`

`Antiallergika - Antiasthmatika - Antitussiva - Bronchodilatatoren - Bronchosekretolytika ~ Mukolytika
`
`
`azelastine, levocabastine or cetirizine showed an effect
`
`on TXB2 production. Cetirizine had also no influence
`on the production of LTC4, and levocabastine only
`slightly inhibited this mediator by 20 % (Fig. 2). Azelas—
`tine and terfenadine, on the other hand, very potently
`decreased LTC4 release by 86 % and even 100 % respec-
`tively.
`
`4. Discussion
`
`The knowledge of allergic rhinitis has been enriched in
`recent years by the investigation of different mediators,
`cells and mechanisms involved in the pathophysiology.
`Certain cytokines, such as TNFo., as well as leukotrienes
`have become matters of further scientific interest.
`
`TNFa is a candidate cytokine relevant to the patho-
`genesis of allergic rhinitis through its capacity for up-
`regulating the expression of endothelial cell adhesion
`molecules, mediating granulocyte chemoattraction,
`and activating eosinophils, mast cells and T cells [27,
`28]. Moreover, TNFO. is localised in nasal mast cells and
`
`is released in rhinitis patients upon allergen challenge
`[29]. TNFa is increased in the nasal epithelial lining
`fluid and in nasal secretions of patients suffering from
`rhinitis [30, 31]. Recently, it has been demonstrated that
`human mast cells and eosinophils are a source of TNFa
`[32, 33].
`
`In the present study we were able to show the anti-
`inflammatory action of azelastine in a human in vitro
`model most suitable to investigate the interaction of
`various cells populations involved in allergic and in-
`flammatory processes. Cells gained from nasal polyps
`include epithelial cells, monocytes, macrophages and
`mast cells as well as infiltrating cells such as lympho-
`cytes, neutrophils and many eosinophils. Upon sensitis-
`ation with human IgE and subsequent stimulation with
`anti-human IgE to mimic allergen contact the cells se-
`crete TNFa. Azelastine, in contrast to levocabastine and
`cetirizine, very potently inhibited the release of TNFCL
`In its effect on TNFa terfenadine was equally effective
`as azelastine.
`
`LTs play an imporant role in nasal congestion, secre-
`tion and cell infiltration, in other words, in the develop-
`ment of the chronic allergic inflammation [34, 35]. In-
`tense production of arachidonic acid metabolites in
`nasal polyp tissue samples has been shown by Jung et
`al. [36]. In agreement with previous data [23], cells avail-
`able in nasal polyps produce LTs and cytokines upon
`allergen stimulation. Recent studies have suggested that
`antihistamines, widely used in the treatment of symp-
`toms of patients with allergic rhinitis, may also possess
`anti-inflammatory properties.
`Loratadine and its active metabolite, descarboxy—
`ethoxyloratadine, help to stabilise mast cells, as evi-
`denced by their ability to inhibit the release of hist-
`amine and LTs [37]. There is evidence that loratadine
`inhibts LT biosynthesis [38]. LTC4 release by peripheral
`blood basophils and tissue mast cells was inhibited by
`
`loratadine in the micromolar range [39, 40]. It has also
`beenlshown that loratadine (0.25—25 umol/l) inhibits
`TNFa—induced LTB4 production in peripheral neutro-
`phil granulocytes [41]. There is no evidence in the liter-
`ature that loratadine might inhibit TNFU. release. How-
`ever, it has been shown that loratadine can inhibit the
`release of IL-6 and IL-8 from human mast cells and en-
`
`dothelial cells to a certain degree [42, 43]. It is believed
`that mizolastine is effective in the treatment of nasal
`
`obstruction due to its inhibitory action on LT syn-
`thesis [38].
`
`Cetirizine substantially reduced the release of LTs
`from blood leukocytes of children with allergic rhinitis
`[44], and in a model of nasal antigen challenge in hu-
`mans it significantly decreased LTC4 levels [45]. How-
`ever, the data obtained in humans are contradictory. In
`a model of allergen—induced early phase for instance
`cetirizine did not influence the elevations of nasal LTC4
`levels [46].
`In previous experiments, azelastine has been shown
`to inhibit the Ca2+-ionophore A23187-induced LT re-
`lease with lC50 values between 1.1 and 36.5 umol/l de-
`pending on animal species and cells used [15, 16, 47-
`49]. Various studies on actively or passively sensitized
`lung preparations of human or guinea-pig origin
`showed that azelastine potently inhibits release of cys-
`teinyl-LTs after allergen or anti-IgE challenge with IC5p
`values in the range of 14 urnol/l to 272 umol/l and was
`superior to other antiallergic/antiasthmatic drugs (i.e.
`ketotifen, terfenadine, DSCG) [4, 12, 47, 50]. The potent
`inhibition by azelastine may be partly a result of the
`inhibition of 5—LOX (5-lipoxygenase), since in rat baso-
`philic leukaemia cell homogenate 5-HETE (12(S]-—
`hydroxy-5- cis-8- cis- 10-trans-14-ciseicosatetraenic acid)
`formation was inhibited by azelastine [49]. Studies on
`isolated murine peritoneal cells showed that azelastine
`significantly inhibited 5-LOX activity (IC50 = 11 umol/l),
`but not LTC4-synthease or LTA4—hydrolase activity [14].
`Furthermore, azelastine exerted little effect on 12—LOX
`
`activity and did not influence COX activity and TXA;_-
`synthesis in human platelets and guinea pig lung
`homogenates [12, 48]. Chand and Sofia suggested that
`azelastine may inhibit the Ca2+-dependent transloca-
`tion of 5-LOX from cytosol to the nuclear envelope or
`it might be a FLAP (5-lipoxygenase activating protein)
`inhibitor rather than directly effecting 5-LOX [51].
`Others indicate that azelastine reduces LT production
`by inhibiting PLA2 and LTC4 synthase [52].
`After a single treatment with azelastine, there was a
`significant reduction in the median level of cysteinyl—
`LT concentration in patients suffering from rhinitis or
`patients who underwent nasal challenge [53]. In this
`study we were able to confirm this anti-inflammatory
`action of azelastine on human nasal polyp cells. Azelas—
`tine (10 umol/l) almost completely abolished the re-
`lease of LTC4 from polyp cells after allergic stimulation.
`TXB2 production, on the other hand, was not affected.
`Hence azelastine, as has been shown for other cell
`
`100 Kiisters et al. — Antihistamines
`
`000004
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`Arzneim.-Forsch./Drug Res. 52, No. 2, 97-102 (2002)
`© ECV - Editio Cantor Verlag, Aulendorf (Germany)
`
`
`
`

`

`
`
`
`
`40]. It has also
`mol/1) inhibits
`
`pheral neutro-
`.nce in the liter-
`or release. How-
`can inhibit the
`1st cells and en-
`
`3]. It is believed
`ttment of nasal
`
`on on LT syn-
`
`release of LTs
`
`allergic rhinitis
`hallenge in hu-
`vels [45]. How-
`
`ontradictory. In
`se for instance
`
`is of nasal LTC4
`
`tas been shown
`induced LT re-
`
`36.5 umol/l de-
`;ed [15, 16, 47-
`
`ively sensitized
`nea—pig origin
`; release of cys-
`lenge with 1C5p
`imol/l and was
`
`tatic drugs (i.e.
`50]. The potent
`a result of the
`ICE? in rat baso-
`
`—HETE (12(S)—
`atetraenic acid)
`49]. Studies on
`that azelastine
`
`50 : 11 pmol/1),
`se activity [14].
`feet on 12—LOX
`
`vity and TXA2—
`.inea pig lung
`suggested that
`ient transloca—
`
`~ar envelope or
`vating protein)
`g 5-LOX [51].
`LT production
`2].
`1e, there was a
`
`el of cysteinyl-
`rom rhinitis or
`
`;e [53]. In this
`‘L-inflammatory
`p cells. Azelas—
`)lished the re-
`
`:ic stimulation.
`LS not affected.
`for other cell
`
`No. 2, 97-102 (2002)
`Kulendorf (Germany)
`
`Arzneim.—Forsch./Dru Res. 52, No. 2, 97-102 (2002)
`© ECV - Editio Cantor erlag, Aulendorf (Germany)
`
`000005
`000005
`
`Kiisters et al. — Antihistamines
`
`
`
`Antiallergic Drugs - Antiasthmatics ~ Antitussives - Bronchodilators - Bronchosecretogogues - Mucolytics
`
`types, specifically inhibits the linear pathway of arachi-
`donic acid metabolism in our cells culture system prob-
`ably by influence on the 5-LOX or possibly on PLA2
`(phospholipase A2) or LTC4 synthase, while leaving the
`cyclic pathway unaffected. Terfenadine in comparison
`to azelastine proved to be more potent in inhibiting
`both LTC4 and TXB2. Although TXB2 release was not as
`strongly decreased as LTC4, terfenadine still is not as
`specific for the linear arachidonic acid pathway. This
`specificity seems to be a unique feature of azelastine.
`However, the exact mechanism of the anti-inflamma-
`
`tory property of azelastine remains to be elucidated.
`In summary, the present study shows the anti-in-
`flammatory action of azelastine in therapeutically rele-
`vant concentrations as assessed by its ability to reduce
`TNFa release as well as inhibit LTC4 production in aller-
`gically stimulated human nasal polyp cells.
`
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`
`Acknowledgements
`The authors wish to thank the manufacturers of levocabastine,
`
`for
`respectively,
`terfenadine and cetirizine hydrochloride,
`kindly providing these drugs. This study was supported by the
`Sachsische Aufbau Bank (Dresden, Germany), grant number
`5476.
`
`Correspondence:
`Dr. Istvan Szelenyi,
`Arzneimittelwerk Dresden GmbH,
`Meissnerstr. 191,
`
`01445 Radebeul (Germany)
`E—mail: stefan.szelenyi@astamedica.de
`
`
`
`1
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`Kiisters et al. — Antihistamines
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`Arzneim.-Forsch.lDrug Res. 52, No. 2, 97-102 (2002)
`© ECV A Editio Cantor Verlag, Aulendorf (Germany)
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