PHARMACY LIBRARY
`UNIVERSITY OF WISCONSIN
`FEB 1 9 2002
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`Madison, WI 53705
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`I
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`Biotechnology in Drug Research
`
`PHARMACY LIBRARY
`UNIVERSITY OF WISCONSIN
`FEB 1 9 2002
`
`Madison, WI 53705
`
`2/ 2002
`
`< en Arzneimittel
`~& Forschung
`b b.O Drug Research
`s _p_e_c_i_al_S~e_c_t1_·o_n _______________ _
`=:::::s
`I
`ECl
`Q.)
`C:
`t:::!
`c:::(
`
`Vol. 52 • No. 2 • pages 73-144 (2002)
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`

`

`■ Antiallergika • Antiasthmatika • Antitussiva ·
`Bronchodilatatoren • Bronchosekretolytika · Mukolytika
`■ Antiallergic Drugs • Antiasthmatics · Antitussives ·
`Bronchodi la tors • Bronchosecretogogues • Mucolytics
`
`73
`
`97
`
`E
`
`Kilsters, S., Schuligoi, R., Hilttenbrink, K.-B.,
`Rudert, J., Wachs, A., Szelenyi, I., Peskar, B. A.
`Wirkung von Antihistaminika auf die Freisetzung
`von Leukotrien und Cytokin aus dispergierten
`Zellen nasaler Polypen
`Effects of Antihistamines on Leukotriene and
`Cytokine Release from Dispersed Nasal Polyp
`Cells
`
`■ Antibiotika • Chemotherapeutika • Virustatika ·
`Zytostatika
`■ Antibiotics • Antiviral Drugs · Chemotherapeutics ·
`Cytostatics
`·
`
`l03
`
`l09
`
`Pandeya, S. N., Yogeeswari, P., Sausville, E. A., E
`Mauger, A. B., Narayanan, V. L.
`Synthese und tumorhemmende Evaluierung von
`4-Bromphenyl-semicarbazon-Derivaten
`Synthesis and Anti tumour Evaluation of 4-Bromo(cid:173)
`phenyl Semicarbazones
`
`Braga, P. C., Dal Sasso, M.
`E
`EinfluB subinhibitorischer Konzentrationen von
`Gatifloxacin auf die Hemmung der Adharenz
`von Staphylococcus aureus und Escherichia coli
`Effects of Sub-minimum Inhibitory Concentra-
`tions of Gatifloxacin on the Inhibition Staphylo-
`coccus aureus and Escherichia coli Adherence
`
`■ Obersicht
`■ Review
`
`G
`Wehling, M.
`~-Blocker und ihre Bedeutung fur die Pharmako(cid:173)
`therapie kardiovaskularer Erkrankungen im Ver-
`gleich
`Comparing P-Blockers and their Relevance in the
`Pharmacotherapy of Cardiovascular Diseases
`
`■ Hypnotika • Psychopharmaka · Sedativa ·
`ZNS~ Th era peuti ka
`■ CNS-active Drugs • Hypnotics • Psychotropics ·
`Sedatives
`
`E
`
`81
`
`Arisawa, H., Fukui, K., Fujise, N.,
`Masunaga, H.
`Allgemeines pharmakologisches Profil des neu(cid:173)
`artigen Muskarinrezeptor-Agonisten SNI-2011
`zur Behandlung von Xerostomie bei Sjt:igren(cid:173)
`Syndrom / 2. Mitteilung: Wirkungen auf das so(cid:173)
`matische und autonome Nervensystem sowie
`die glatte Muskulatur
`General Pharmacological Profile of the Novel
`Muscarinic Receptor Agonist SNI-2011, a Drug
`for Xerostomia in Sjogren's Syndrome I 2nd Com(cid:173)
`munication: Effects on somatic nervous system
`and on autonomic nervous system and smooth
`muscle
`
`■ Herz-Kreislauf-Therapeutika · Kardiaka ·
`Koronar-Thera peutika
`■ Cardiac Drugs • Cardiac Stimulants ·
`Coronary Drugs
`
`E
`
`89
`
`Wittig, J., Leipolz, I., Graefe, E. U., Jaki, B.,
`Treutter, D., Veit, M.
`Quantifizierung von Procyanidinen in Cratae(cid:173)
`gus-Extrakte enthaltenden oralen Fertigarznei(cid:173)
`mitteln
`Quantification of Procyanidins in Oral Herbal
`Medicinal Products Containing Extracts of Crat(cid:173)
`aegus Species
`
`E = Publication in English. G = Publication in German.
`
`

`

`113
`
`■ Buchbesprechungen
`■ Book Reviews
`
`137
`
`■ Biotechnology in Drug Research:
`Scientific Centers / Companies in Biobusiness
`
`BioRegioN - Biotechnology in Lower Saxony E 140
`
`E
`
`Ortega, M. A., Sainz, Y., Montoya, M. E.,
`Jaso, A., Zarranz, B., Aldana, I., Monge, A.
`Derivate van Chinoxalin-2-carbonitril und
`Chinoxalin-2-carbonitril-1,4-di-N-oxid mit
`Wirkung gegen Mycobacterium tuberculosis
`Anti-Mycobacterium tuberculosis Agents Derived·
`from Quinoxaline-2-carbonitrile and Quinoxa(cid:173)
`line-2-carbonitrile 1,4-di-N-oxide
`
`C. F. Gomes, D. de, Alegria, L. V., Freire de
`Lima, M. E., Leon, L. L., Araujo, C. A. C.
`Synthetische Derivate von Curcumin und ihre
`Wirksamkeit gegen Leishmania amazonensis
`Synthetic Derivatives of Curcumin and their
`Activity against Leishmania amazonensis
`
`E 120
`
`■ Besondere Themen
`■ Special Themes
`
`Yunes, R. A., Heinzen, V. E. F., Filho, V. C.,
`Lazzarotto, M.
`Von der manuellen Topliss-Methode hin zu
`einer rnodifizierten quantitativen Methode
`From the Manual Method of Topliss to a
`Modified Quantitative Method
`
`E 125
`
`■ Patentinformationen
`■ Information on Patents
`
`E 133
`
`Habernickel, V. J.
`Der Pharrnamarkt im Spiegel der Patente I
`Anxiolytika und Antidepressiva / Wirkstoffe zur
`Behandlung kognitiver Storungen / Antibakte-
`rielle Substanzen / Antithrornbotika
`The Pharma Market as Reflected by Patents I
`Agents for the treatment of anxiety and depres(cid:173)
`sion I Agents for the treatment of e.g. cognitive
`disorders I Compounds having antibacterial
`activity I Agents for the treatment of thrombo(cid:173)
`embolic disorders
`
`

`

`Antiallergika · Antiasthmatika • Antitussiva · Bronchodilatatoren
`Bronchosekretolytika • Mukolytika
`
`Arz n e i m Forsch Drug Res:
`
`Antiallergic Drugs · Antiasthmatics • Antitussives • Bronchodilators
`Bronchosecretogogues • Mucolytics
`
`This material may be protected by Copyright law (Title 17 U.S. Code)
`
`Effects of Antihistamines on
`Leukotriene and Cytokine Release
`from Dispersed Nasal Polyp Cells
`
`Sabine Kiistersa, Rufina Schuligoih, Karl-Bernd Hiittenbrinkc, Jutta Ruderta, Angela Wachsa,
`Istvan Szelenyi a, and Bernhard A. Peskarh
`
`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)
`
`control or mean IC50 (n = 5). Azelastine
`and terfenadine inhibited TNFa release
`with IC50 values of 6.2 µmol/1 and 4.3
`µmol/1, respectively. Terfenadine reduced
`TXB2 release by 37 ± 15 %, and LTC4 re(cid:173)
`lease was decreased by azelastine and ter(cid:173)
`fenadine very potently by 86 % and
`100 %, respectively. Azelastine shows
`anti-inflammatory properties in thera(cid:173)
`peutically relevant concentrations as as(cid:173)
`sessed by its ability to reduce TNFa re(cid:173)
`lease as well as its ability to inhibit LTC4
`production in allergically stimulated hu(cid:173)
`man nasal polyp cells.
`
`Summary
`
`In this study the effects of antihistamines
`on the release of eicosanoids and the
`pro-inflammatory cytokine tumor necro(cid:173)
`sis factor alpha (TNFa) were compared.
`Enzymatically dispersed cells from hu(cid:173)
`man nasal polyps served as an in vitro
`model of chronic respiratory mucosa! in(cid:173)
`flammation. Nasal polyp cells (2 x 106/
`ml) were sensitized with human IgE pre(cid:173)
`incubated° with azelastine (CAS 58581-89-
`8), terfenadine (CAS 50679-08-8), levocab(cid:173)
`astine (CAS 79516-68-0) or cetirizine
`(CAS 83881-51-0), and stimulated with
`anti-human immunoglobulin E (lgE).
`Thromboxane B2 (TBX2 ) and leukotriene
`C1 (LTC4) were measured by radio(cid:173)
`immunoassay (RIA), TNFa by enzyme-lin(cid:173)
`ked immunosorbent assay (ELISA). Data
`represent mean values of % inhibition
`estimated from the untreated positive
`
`Zusammenfassung
`
`Wirkung 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
`
`entziindungsfcirdernden Cytokins ,,Tu(cid:173)
`mor necrosis factor" a (TNFa) unter(cid:173)
`sucht. Enzymatisch dispergierte Zellen
`von menschlichen nasalen Polypen dien(cid:173)
`ten als In-vitro-Modell der chronischen
`mukosalen Entziindung der Atemwege.
`Diese Zellen 2 x 106/ml) wurden mit
`
`Keywords
`
`• Azelastine
`■ Cetirizine
`• Leukotriene
`■ Levocabastine
`■ Nasal polyps, human
`■ Terfenadine
`■ 1\unor necrosis factor a
`
`Arzneirn.-Forsch./Drug Res.
`52, No. 2, 97-102 (2002)
`
`

`

`menschlichem Immunglobulin E (lgE)
`sensibilisiert. AnschlieRend wurden sie
`mit Azelastin (CAS 58581-89-8), Terfena(cid:173)
`din (CAS 50679-08-8), Levocabastin (CAS
`79516-68-0) oder Cetirizin (CAS 83881-
`51-0) vorinkubiert. Die allergische Stimu(cid:173)
`lation erfolgte mit Anti-human-IgE.
`Thromboxan B2 (TXB2) und Leukotrien
`C4 (LTC4) wurden mit Hilfe von RIA (ra(cid:173)
`dioimmunoassay), TNFa mit ELISA (en-
`
`zyme-linked immunosorbent assay) ge(cid:173)
`messen. Die Ergebnisse entsprechen ei(cid:173)
`ner prozentualen Hemmung im Vergleich
`zu nicht-behandelten Positivkontrollwer(cid:173)
`ten (n = 5). Azelastin und Terfenadin
`hemmten die TNFa-Freisetzung mit IC50-
`Werten von 6,2 µmol/1 bzw. 4,3 µmol/1.
`Terfenadin reduzierte die TXBrFreiset(cid:173)
`zung um 37 + 15 %. Die LTC4-Freisetzung
`wurde durch Terfenadin und Azelastin
`
`sehr stark, um 86 % bzw. 100 % ge(cid:173)
`hemmt. In therapeutisch relevanten Kon(cid:173)
`zentrationen besitzt Azelastin entziin(cid:173)
`dungshemmende Eigenschaften, die
`durch die Hemmung der TNFa- bzw. der
`LTC4-Freisetzung charakterisiert werden
`konnten.
`
`l,
`!.
`
`I. Introduction
`Histamine Hi-receptor antagonists together with top(cid:173)
`ical steroids are the treatment of choice in allergic rhin(cid:173)
`itis. The phthalazinone derivative azelastine has suc(cid:173)
`cessfully been introduced in the therapy of allergic rhin(cid:173)
`itis and conjunctivitis. It has also been demonstrated
`that azelastine reduces nasal congestion [1-3]. This fea(cid:173)
`ture, which disginguishes azelastine from oral antihis(cid:173)
`tamines, is of great interest, because corticosteroids are
`known to be quite effective in relieving nasal conges(cid:173)
`tion, whereas antihistamines are effective for the sneez(cid:173)
`ing, itching, watery secretion, but not congestion.
`It is well known that azelastine and some other anti(cid:173)
`histamines may possess properties unrelated to block(cid:173)
`ade of histamine at its receptor level. In animal studies,
`it has been demonstrated that in addition to acting as
`a histamine H1 -receptor antagonist, azelastine also in(cid:173)
`hibits the production of many chemical mediators of
`the allergic response [4-9]. Azelastine has been re(cid:173)
`ported to inhibit leukotriene (LT)-mediated acute aller(cid:173)
`gic bronchoconstriction in guinea pigs and passive cu(cid:173)
`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(cid:173)
`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(cid:173)
`ing human cells [4, 13-16]. Moreover, polypous tissue
`samples have been shown to produce more cysteinyl(cid:173)
`LT upon stimulation than normal nasal mucosa indicat(cid:173)
`ing an altered pattern of arachidonic acid pathways
`[17]. Therefore, the present study has been performed
`with enzymatically dispersed cells obtained from hu(cid:173)
`man nasal polyps to compare the effect of antihistam(cid:173)
`ines on the release of pro-inflammatory cytokines and
`eicosanoids. Nasal polyps are a model of chronic respir(cid:173)
`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(cid:173)
`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 [19-22].
`
`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(cid:173)
`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 diges·tion according to Campbell
`and Bousquet (23]. Briefly, the minced polyps were incubated
`for 2 h at 37 °C in RPMI 1640 (1 g tissue per 4 ml) containing
`2.0 mg/ml protease, 1.5 mg/ml collagenase, 0.75 mg/ml hyalu(cid:173)
`ronidase and 0.05 mg/ml DNase (all enzymes Sigma, Deisen(cid:173)
`hofen, Germany). Then, the cell suspension was :filtered to re(cid:173)
`move any undigested tissue and the cells were washed 3 times.
`After an incubation in the presence of human IgE (1 µg/ml in
`RPMI 1640 10 % fetal calf serum CFCS), Biochrom) for 1 h at
`37 °C, the cells were washed two times, counted and resuspen(cid:173)
`ded at a concentration' of 2 x 106 cells/ml. Then 1 µg anti-IgE
`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(cid:173)
`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(cid:173)
`many). Terfenadine
`(CAS 50679-08)
`levocabastine hydro(cid:173)
`chloride (CAS 7954 7-78-7) and cetirizine hydrochloride (CAS
`83881-~2-l) were kindly provided/by the respective manufac:
`turers.
`
`2.3. Cytokine determination
`To determine the effect of antihistamines on cytokine release
`nasal polyp cells were pre-incubated with 3, 10 and 30 µmoll!
`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(cid:173)
`pernatant was stored at -70 °c until determination of TNFa
`contents by sandwich ELISA using an antibody pair and stand(cid:173)
`ard protein from PharMingen (Hamburg, Germany).
`
`9 8 Ktisters et al. - Antihistamines
`
`Arzneim.-Forsch./Drug Res. 52, No. 21 97-102 (2002)
`© ECV '. Editio Cantor Verlag, Aulendorf (Germany)
`
`

`

`Antiallergic Drugs · Antiasthmatics • Antitussives • Bronchodilators • Bronchosecretogogues . Mucolytics
`
`2.4. Eicosanoid determination
`To determine the effect of antihistamines on eicosanoid release
`nasal polyp cells were pre-incubated with 10 µmol/1 of either
`azelastine, terfenadine, cetirizine and lcvocabastine respec(cid:173)
`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(cid:173)
`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, 11, 12, 14, 15(n) 3H)TXB2 and [14, 15, 19, 20-3H (n))LTC4
`(both from New England Nuclear, Vienna, Austria) as radio(cid:173)
`ligands and synthetic TXB 2 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 ± SD. The IC50 values were
`determined using an in-house programmed PC software pack(cid:173)
`age (EDX 2.1) .
`
`3. Results
`3.1. Effect of antihistamines on TNFa release
`from nasal polyp cells
`The finding that azelastine may act via mechanisms dif(cid:173)
`ferent from blocking the histamine receptor prompted
`us to investigate its anti-inflammatory potency. There(cid:173)
`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 IgE, served as positive control and re(cid:173)
`leased 400 to 500 pg/ml TNFa into the supernatant 18 h
`after addition of the stimulus. Azelastine and terfenad(cid:173)
`ine inhibited the release of TNFa concentration-de(cid:173)
`pendently and were very potent in this respect, since
`they almost completely prevented TNFa release at 30
`µmol/1 (Fig. 1). Both antihistamines were equally effec(cid:173)
`tive with an IC50 value of 6.2 µmol/1 for terfenadine and
`4.3 µmol/1 for azelastine. At a concentration of 3 µmol/1
`levocabastine and cetirizine were as effective as terfen(cid:173)
`adine in inhibiting TNFa release by 40 % and 35 %, re(cid:173)
`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(cid:173)
`tion of arachidonic acid metabolites. To investigate
`whether the cyclic or the linear pathway of arachidonic
`acid metabolism was influenced, we determined TXB2
`
`120
`
`100
`
`.,
`., ..
`.,
`~~
`ti 0
`u...!:;
`z C:
`f- 0
`... u
`§~
`:;::. ......
`:E :c
`·=
`
`------ azelastlne
`~ terlenadlne
`----T- lavocabastine
`-v- cetirizlne
`
`80
`
`60
`
`40
`
`20
`
`!
`
`10
`
`3
`
`µmol/1
`
`T
`
`I
`
`30
`
`Fig. 1: Inhibition of allergically induced release ofTNFa from en(cid:173)
`zymatically dispersed nasal polyp cells. Nasal polyp cells (2 x 106/
`ml) were sensitized with human IgE for 1 h. 30 min prior to
`stimulation with anti-human IgE 3, 10, or 30 µmol/1 azelastine,
`terfenadine, levocabastine and cetirizine were added respectively.
`18 h after anti-IgE addition supernatants were removed and
`frozen at - 70 °C until cytokine determination with ELISA. Data
`represent mean values of % inhibition estimated from the un(cid:173)
`treated positive control ± SD (n = 5)
`
`and LTC4 in parallel. Positive control nasal polyp cells,
`which were only sensitized with human IgE and stimu(cid:173)
`lated with anti-human IgE, released 320 pg/ml TXB2
`and 350 pg/ml LTC4 • Azelastine, terfenadine, levocabas(cid:173)
`tine or cetirizine were given to the nasal polyp cells at
`a concentration of 10 µmol/1 30 min before addition of
`the allergic stimulus anti-IgE. Of all the antihistamines
`investigated only terfenadine reduced TXB2 release by
`37 ± 15 % as compared to the positive control assessed
`one hour after addition of anti-IgE. On average neither
`
`1
`
`120
`
`100
`
`., ., .. .,
`
`~ 80
`o·
`f-
`..J
`0
`C:
`0
`:;::
`
`60
`
`:E :c
`.!:
`".'!-
`
`40
`
`20
`
`0
`
`I
`
`I
`
`azelastine
`
`tarfanadlna
`
`levocabastlne
`
`Fig. 2: Inhibition of allergically i.nduced release of LTC., from. en(cid:173)
`zymatically dispersed nasal polyp cells. Nasal polyp cells (2 x 106
`/
`ml) we:re sensitized with human IgE for I h. Thirty minutes prior
`to stimulation with anti-human IgE 10 µmol/1 azelastine, terfena(cid:173)
`dine and levocabastine were added respectively. One hour after
`anti-JgE addition supernatanls were removed, shock frozen in li(cid:173)
`quid nitrogen aJJd slored at - 70 °0 untll LTC, determination.
`Data represent mean values of % Inhibition estimated Crom the
`untreated positive control ± SD (n = 5).
`
`

`

`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(cid:173)
`tine and terfenadine, on the other hand, very potently
`decreased LTC4 release by 86 % and even 100 % respec(cid:173)
`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 TNFa, as well as leukotrienes
`have become matters of further scientific interest.
`TNFa is a candidate cytokine relevant to the patho(cid:173)
`genesis of allergic rhinitis through its capacity for up(cid:173)
`regulating the expression of endothelial cell adhesion
`molecules, mediating granulocyte chemoattraction,
`and activating eosinophils, mast cells and T cells [27,
`28]. Moreover, TNFa 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(cid:173)
`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(cid:173)
`flammatory processes. Cells gained from nasal polyps
`include epithelial cells, monocytes, macrophages and
`mast cells as well as infiltrating cells such as lympho(cid:173)
`cytes, neutrophils and many eosinophils. Upon sensitis(cid:173)
`ation with human IgE and subsequent stimulation with
`anti-human IgE to mimic allergen contact the cells se(cid:173)
`crete TNFa. Azelastine, in contrast to levocabastine and
`cetirizine, very potently inhibited the release of TNFa.
`In its effect on TNFa terfenadine was equally effective
`as azelastine.
`LTs play an imporant role in nasal congestion, secre(cid:173)
`tion and cell infiltration, in other words, in the develop(cid:173)
`ment of the chronic allergic inflammation [34, 35]. In(cid:173)
`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(cid:173)
`able in nasal polyps produce LTs and cytokines upon
`allergen stimulation. Recent studies have suggested that
`antihistamines, widely used in the treatment of symp(cid:173)
`toms of patients with allergic rhinitis, may also possess
`anti-inflammatory properties.
`Loratadine and its active metabolite, descarboxy(cid:173)
`ethoxyloratadine, help to stabilise mast cells, as evi(cid:173)
`denced by their ability to inhibit the release of hist(cid:173)
`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
`been shown that loratadine (0.25-25 µmol/1) inhibits
`TNFa-induced LTB4 production in peripheral neutro(cid:173)
`phil granulocytes [41]. There is no evidence in the liter(cid:173)
`ature that loratadine might inhibit TNFa release. How(cid:173)
`ever, it has been shown that loratadine can inhibit the
`release of IL-6 and IL-8 from human mast cells and en(cid:173)
`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(cid:173)
`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(cid:173)
`mans it significantly decreased LTC4 levels [45]. How(cid:173)
`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(cid:173)
`lease with IC50 values between 1.1 and 36.5 µmol/1 de(cid:173)
`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(cid:173)
`teinyl-LTs after allergen or anti-IgE challenge with IC5p
`values in the range of 14 µmol/1 to 272 µmol/1 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(cid:173)
`philic leukaemia cell homogenate 5-HETE (12(S)(cid:173)
`hydroxy-5-cis-8-cis-l 0-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 µmol/1),
`but not LTC4-synthease or LTA.i-hydrolase activity [14].
`Furthermore, azelastine exerted little effect on 12-LOX
`activity and did not influence COX activity and TXA2 -
`synthesis in human platelets and guinea pig lung
`homogenates [12, 48]. Chand and Sofia suggested that
`azelastine may inhibit the Ca2+-dependent transloca(cid:173)
`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(cid:173)
`LT concentration in patients suffering from rhinitis or
`patients who underwent nasal challenge [53]. I:q. this
`study we were able to confirm this anti-inflammatory
`action of azelastine on human nasal polyp cells. Azelas(cid:173)
`tine (10 µmol/1) almost completely abolished the re(cid:173)
`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
`
`10 Q Kiisters et al - Antihistamines
`
`Arzneim.-Forsch./Drug Res. 52, No. 2, 97- 102 (2002)
`© ECV • Editio Cantor Verlag, Aulcndorf (Germany)
`
`

`

`Antiallergic Drugs • Antiasthmatics • Antitussives • Bronchodilators • Bronchosecretogogues • Mucolytics
`
`types, specifically inhibits the linear pathway of arachi(cid:173)
`donic acid metabolism in our cells culture system prob(cid:173)
`ably by influence on the 5-LOX or possibly on PIA2
`(ph~spholipase 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 ci unique feature of azelastine.
`However, the exact mechanism of the anti-inflamma(cid:173)
`tory property of azelastine remains to be elucidated.
`In summary, the present study shows the anti-in(cid:173)
`flammatory action of azelastine in therapeutically rele(cid:173)
`vant concentrations as assessed by its ability to reduce
`TNFu release as well as inhibit LTC4 production in aller(cid:173)
`gically stimulated human nasal polyp cells.
`
`5. References
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`macodynamic and pharmacokinetic properties, and thera(cid:173)
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