European Journal of Pharmaceutical Sciences 17S (2002) S77–S85www.elsevier.nl / locate/ejps

E UFEPS 2002 Supplement

PO-92 CHLAMYDIA PNEUMONIAE REPLICATION [1] Leinonen M, Saikku P. Evidence for infectious agentsAND INFECTIOUS PROGENY PRODUCTION IN DIF- in cardiovascular disease and atherosclerosis. LancetFERENT CELL LINES Infect Dis. 2002;2:11–7.

1 1 2 3J Alvesalo , P Tammela , H Vuorela , M Leinonen , P4 a 1Saikku & P Vuorela – Viikki Drug Discovery Technol-

ogy Center, Department of Pharmacy, P.O. Box 56, FIN- PO-93 THE CINOD AZD3582 iNHIBITS CYCLOOXY-200014 University of Helsinki, Finland; Division of Phar- GENASE AND REDUCES PAIN, INFLAMMATION

macognosy, Department of Pharmacy, University of Hel- AND FEVER IN THE RAT3 1 1 1sinki, Finland; National Public Health Institute, Depart- Odd-Geir Berge , Johan Raud , Janet Hoogstraate & John

4 2 1ment in Oulu, Finland; Department of Medical Micro- ¨ ¨L Wallace – AstraZeneca R&D Sodertalje, S-151 852biology, University of Oulu, Finland ¨ ¨Sodertalje, Sweden; University of Calgary, Calgary,

Alberta, T2N 4N1, Canada

Chlamydia pneumoniae is a common human respiratorypathogen causing pneumonia, bronchitis, sinusitis and The objective of this study was to assess the analgesic,pharyngitis worldwide. In addition, chronicC. pneumoniae anti-inflammatory and anti-pyretic efficacy of AZD3582,infection has been associated with coronary heart disease the first of a new class of COX-inhibiting nitric oxideand acute myocardial infarction [1]. donators (CINODs) that act through balanced inhibition of

C. pneumoniae is an obligatory intracellular parasite COX-1 and COX-2 isoenzymes and nitric oxide donation.requiring living cells for its growth. For this reason it is AZD3582 [4-(nitrooxy)butyl-(2S)-2-(6-methoxy-2-important to find cell lines that are capable of ‘‘hosting’’C. naphthyl)propanoate] or naproxen at equimolar doses (0.3,pneumoniae growth cycle and can thus be used forC. 1, 3, 10, 30 and 100mmol /kg), or vehicle (n57–8 perpneumoniae culture and drug screening purposes. group) were administered orally 1 h before injection of

In this study we investigated the ability ofC. pneumo- carrageenan into the ankle joint of male Sprague–Dawleyniae to infect, replicate and produce infectious progenies in rats; pain was scored using a 4-point scale (05full weight-five different cell lines of human (HL, Calu-3, Caco-2), rat bearing, 35paw completely elevated). Anti-inflammatory(GH C and mouse (GT1-7) origin. Cells were grown in efficacy was assessed by giving the compounds (doses as4 1)

324-well plates and infected with 10 IFU (inclusion-form- above,n55–6 per dose; vehicle,n512) to male Wistaring units) of C. pneumoniae elementary bodies. Formed rats 30 min before carrageenan injection into the hindpawinclusions were counted 72 h after infection from cover- footpad. Fever was induced in Sprague–Dawley rats byslips stained with fluorescein-conjugated monoclonal anti- subcutaneous injection of Brewer’s yeast; rectal tempera-

body to chlamydial lipopolysaccharide (Pathfinder ). In- tures were measured before and 2 h after administration of`fectious progeny production was tested from replicate AZD3582, naproxen (0.3, 1, 3, 10 and 30 ımol /kg) or

wells. In this test, cells were scraped and 200ml of the vehicle (n58 per group). COX-1 and COX-2 inhibitorysuspension was used to infect HL-cells, which are routine- activity of AZD3582 and rofecoxib, a COX-2 selectively used in our laboratory to growC. pneumoniae. NSAID, were compared using an airpouch model withProgenies were considered infectious, if inclusions could male Wistar rats (n55 per group). Air (20 ml) was injectedbe seen in HL cell cultures. (s.c.) on the back followed 2 and 5 days later by further

C. pneumoniae was shown to be capable to infect, injections (10 ml) at the same site. All injections werereplicate and produce infectious progenies in tested cell performed under anaesthesia. Carrageenan was injectedlines. However, none of the cell lines tested was better into the airpouch 24 h after the last air injection. Thethan HL-cell line. IFU numbers in tested cell lines ranged pouch was incised under anaesthesia 6 h later. AZD3582from 10% to 30% compared to those in HL-cells. All (3, 15 or 30mmol /kg), rofecoxib (0.1, 0.3, 1 or 3 mg/kg)tested cell-lines were capable of producing viable or vehicle was given orally 1 h prior to carrageenan.progenies. Exudate samples were used to quantify leukocyte con-

0928-0987/02/$ – see front matter 2002 Published by Elsevier Science B.V.PI I : S0928-0987( 02 )00157-4

S78 EUFEPS 2002 Supplement 17S (2002) S77–S85

centration, characterize the cellular infiltrate and measure histological alterations significantly improve compared toPGE concentration (an index of COX-2 activity). Whole control.2

blood thromboxane synthesis was measured in venous For modeling the human stroke, a rat transient focalblood as an index of COX-1 activity. ischaemia model was applied in our laboratory. The

AZD3582 and naproxen dose-dependently reduced pain transient occlusion of middle cerebral artery with anbehaviour. At 5 h post-injection, the ED for AZD3582 intraluminal nylon suture embolus induced severe is-50

`(2.9 ımol /kg) was lower than that for naproxen (9.2 chaemia and necrosis in brain areas supplied by middlemmol /kg) while similar potencies were obtained at 3 h cerebral artery. The AMPA antagonist 2,3-benzodiazepines(1.2 vs. 1.0mmol /kg). AZD3582 and naproxen dose- show marked neuroprotection even after delayed treatment.dependently reduced paw oedema. Both agents showed These results suggest that AMPA receptors play an im-significant anti-inflammatory effect at 10mmol /kg with portant role in the development of neuronal damage ofsimilar ED values (16mmol /kg). Fever induced by reperfusion. Talampanel and analogues have great po-30

Brewer’s yeast was reduced dose-dependently by both tential clinical utility as a therapeutic agent in acute strokecompounds; the lowest effective dose was 3mmol /kg for in humans.

`AZD3582 and 1 ımol /kg for naproxen. Plasma monitoring´ ´showed a relative oral bioavailability of 50–70% for [1] Tarnawa I, Farkas S, Berzsenyi P, Pataki A, Andrasi F.

AZD3582 vs. naproxen. In the airpouch model, AZD3582 Electrophysiological studies with a 2,3-benzodiazepinesignificantly reduced PGE content at 15mmol /kg, whole muscle relaxant: GYKI 52466. Eur. J. Pharmacol.,2

blood TXB synthesis at 3mmol /kg and leukocyte 1989;167:193–199.2

´ ´infiltration at 15 mmol /kg (corresponding results for [2] Berzsenyi P, Hamori T, Botka P, Tarnawa I, Andrasi F.rofecoxib were 3 mg/kg, ineffective at highest dose tested, Anticonvulsant and myorelaxant activity of the deriva-and 1 mg/kg, respectively). tives of GYKI 52466. Eur. J. Neurosci., 1992;S5:2191.

The balanced COX inhibition by AZD3582 results indose-dependent reductions in pain, inflammation and feverin animals. The overall efficacy of AZD3582 is at least PO-95 ANTITUMOUR EFFICACY OF THE AL-equal to that of naproxen. The clinical efficacy of LERGEN-REMOVED EXTRACT (ACM909Q) IN RHUSAZD3582 and the ability of its multi-pathway mechanism VERNICIFLUA

1 2 2of action to protect the GI system are being investigated in Choi WonCheol , Z Sminova , I Kubasova & A2 1man. Baryshnikov – K.H.W. Medical Foundation Cancer

2Research Institute, Inchon, Korea; Russian N.N. BlockinCancer Center, Moscow. Russia

PO-94 NON-COMPETITIVE AMPA ANTAGONISTS INRush verniciflua tree has been used, as an excellent

THE THERAPY OF STROKEmedicine in Oriental countries for about 1,000 years

´ ´ ´P Berzsenyi, H Szabo, K Horvath & F Andrasi – IVAXbecause of it is outstanding anticancer activities. Rush tree

Drug Research Institute, H-1045 Budapest, Berlini u. 47–inducing some allergies has very weaker adverse effect of

49, Hungarymedicine, compared with anticancer chemicals, which havebeen already used. In this abstract is described the anti-

2,3-Benzodiazepines, synthetized at IVAX Drug Research cancer activities of allergen-removed-extract named asInstitute, are glutamate receptor antagonists that selectively ACM909G.inhibit the AMPA receptor subtype [1]. Talampanel, a There were used three- and nine-percent extract con-prominent member of this family, is now under Phase II centrations of ACM909G at oral administration during forclinical trials as an antiepileptic drug. Talampanel and its 5 days. To investigate antitumour properties transplantablenovel analogues inhibit spreading depression induced by breast carcinoma Ca-755 and cancer of uterine cervixAMPA in isolated chicken retina and reverse the AMPA RShM-5 were used. Antitumour effect was estimated byinduced inward currents in whole-cell patch-clamp record- criteries: tumour growth inhibition (TGI%) and increase ofings of acutely isolated neonatal rat Purkinje neurons. In life span (ILS%). TGI was calculated by the formula: 100

3mice and rats they strongly inhibit the convulsions induced (Vc/Vt-1) where Vc mean volume (mm ) of tumours inby electroshock and several chemoconvulsants, and have control group, Vt – mean volume of tumours in treatmentpotent muscle relaxant activity [2]. They also have im- group. ILS was calculated by the formula: 100(T/C-1),portant antinociceptive activity in analgesic models. where T is the mean life span (MLS, days) of treatment in

An animal model for multiple sclerosis, guinea pig group, C is MLS of animals in control group.myelin basic protein induced experimental autoimmune Nine-percent extract of ACM909G in 600 mg/kg doseencephalomyelitis was used for determination of the exerts high antitumour effect causing growth inhibition ofefficacy of our drugs on this type of neurodegenerative RShM-5 to 86%. Antitumour effect of ACM909G wasdisorder. After 8 days treatment the motor disability and maintained during two weeks after treatment. MLS of

EUFEPS 2002 Supplement 17S (2002) S77–S85 S79

treatment animals was increased on 43% in comparison blood–brain barrier permeation from three-dimension-with control mice. Lower antitumour efficacy three-percent al molecular structure. J Med Chem, 2000;43:2204–extract of ACM909G in 300 mg/kg dose possesses on 16.breast carcinoma Ca-755 (TGI555%). ILS of mice withCa-755 in comparison with control animals was notobserved in this regimen of administration [1]. PO-97 CYTOTOXIC EFFECT OF TRANS-RESVERAT-

Thus, the search antitumour properties nine-percent ROL ON HeLa CELL LINEextract of ACM909G on RShM-5 demonstrates perspective B Ergun & D Kocaman – Department of Pharmaceuticalof further investigation antitumour activity of this extract. Toxicology, Faculty of Pharmacy, Anadolu University,

Eskisehir, Turkey[1] Experimental Evaluation of Antitumor Drugs in the

USA and USSR and Clinical Correlations. NCI Mono- Several studies have showed that resveratrol, a phenolicgraph, 1980;55:25–96. compound in flavanoid structure, has anticarcinogenic

activity [1,2]. Red wine contains more resveratrol thanpink and white wines [3]. It was demonstrated that trans-

PO-96 AVOLSURF MODEL OF PROPAFENONE-TYPE resveratrol is more effective than cis-resveratrol [2]. Re-MODULATORS OF MULTIDRUG RESISTANCE sveratrol has showed a dose-dependent effect on inhibitionG Ecker, D Kaiser & P Chiba – Institute of Pharmaceutical of THP-1 (human monocytic leukemia cells) and SCC-25Chemistry, University of Vienna, Althanstraße 14, 1090 (human oral epithelial carcinoma cells) cell lines [4,5].Wien, Austria In this study we investigated the inhibitory effect of

trans-resveratrol on HeLa cervics adenocarcinoma cellOne of the major mechanisms responsible for development line. Trans-resveratrol was administrated in 50mg/ml–of multiple drug resistance in tumour therapy is over- 0.406mg/ml dose range by dilution method and it wasexpression of membrane bound drug transport pumps, such seen that inhibitory activity is non dose-dependent. It wasas P-glycoprotein (PGP). PGP transports a wide variety of found that the highest inhibitory effect is at 50mg/mlstructurally and functionally diverse natural product toxins resveratrol dose (47% inhibition).out of tumour cells, thus leading to decreased accumula-tion of anticancer drugs in the cells. Inhibition of PGP [1] Soleas G J, Diamandis E P, Goldberg D M. Re-leads to reversion of resistance. Additionally, together with sveratrol: A molecule whose time has come? Andthe cytochrom-complex, PGP is one of the key players in gone?. Clinical Biochemistry, 1997;30:91–113.absorption/detoxification pathways. Thus, models for pre- [2] Fremont L. Biological effects of resveratrol. Lifediction of both PGP/substrate and PGP/ inhibitor inter- Sciences, 2000;66:663–73.action represents an interesting approach for early ADME [3] Romero-Perez A I, Lamuela-Raventos R M, Water-profiling of compound libraries. house A L, Torre-Boronat M C. Levels of cis- and

A valuable tool for modelling complex structure-activi- trans-resveratrol and their glucosides in white and rosety /property relationships is the software package VolSurf Vitis vinifera wines from Spain. Journal of Agricultur-(TRIPOS), which is based on the calculation of molecular al Food Chemistry, 1996;44:2124–28.interaction fields. Both predictive models for blood–brain [4] Tsan M, White J E, Maheshwari J G, Bremner T A,and caco-2 permeation were successfully established [1]. Sacco J. Resveratrol induces Fas signalling-indepentUsing a training set of 131 propafenone-type inhibitors of apoptosis in THP-1 human monocytic leukaemia cells.PGP and several combinations of different probes, we were British Journal of Haematology, 2000;109:405–12.able to generate a VolSurf model for prediction of PGP- [5] Elattar T M A,Virji A S. The effect of red wine and its

2inhibitory activity. Q values were in the range of 0.57– components on growth and proliferation of human oral0.69 with the combination of a water probe and a squamous carcinoma cells. Anticancer Research,hydrophobic probe as optimum. Using an external test set 1999;19:5407–14.of 50 propafenone-analogs of our in house library, an rvalue of 0.86 was obtained. This demonstrates the goodexternal predictivity of the model. Further investigations PO-98 THE BENEFICIAL EFFECTS OF GENISTEINwill focus on an extension of the model to other classes of ON THE CIRCULATORY FAILURE INDUCED BYcompounds, such as dihydropyridines, phenothiazines and ENDOTOXIN IN THE RATbenzopyranes. Z Fatehi-Hassanabad & M Fatehi – Department of

We gratefully acknowledge financial support by the Physiology & Pharmacology, Faculty of Medicine, P.O.Austrian Science Fund (P13581-MOB).VolSurf was gener- Box 91775-1843, Mashhad University of Medical Sci-ously provided by TRIPOS. ences, Mashhad, Iran

[1] Crivori P, Cruciani G, Carrupt P-A, Testa B. Predicting The effects of genistein, an inhibitor of protein tyrosine

S80 EUFEPS 2002 Supplement 17S (2002) S77–S85

kinase, and L-NAME (N-nitro-L-arginine methyl ester), an duction of the lesion area biomechanics. A different patterninhibitor of nitric oxide synthase, on endotoxin-induced is found in the contra-lateral intact areas. This may help toshock were investigated in the thiopental anesthetized rats. define other complementary strategies to improve thera-We also studied the effects of endotoxin on the vascon- peutical efficacy on this patients.strictor responses to sympathetic nerve stimulation (SNS)in the rat isolated perfused mesenteric bed. Endotoxin [1] Dobrev H. Use of Cutometer to assess epidermal

21injection (10 mg kg , i.p.) produced a marked hypoten- hydration. Skin Research and Technology,sion and a reduction of the pressor responses elicited by 2000;6:239–244.

21phenylephrine (0.1, 0.3 and 3mg kg , i.v.). Pretreatment [2] Rodrigues, L., Berardesca, E., Gummer, C.L.,21of the rats with either genistein (10 mg kg i.p. 2 h before ´ `Leveque, J.L., Loden, M., Masson, Ph., Parra, J.L.,

21endotoxin injection) or L-NAME (0.1 mg kg , i.p. 30 min ´Pierard, G.E, Rogiers, V., Threvethan, M. EEMCObefore endotoxin injection) and combination of both, Guidance to the in vivo assessment of tensile functionsattenuated the hypotension caused by endotoxin. Sympa- of the skin. Part 2: instrumentation and test modes,thetic nerve stimulation (SNS) caused a frequency-depen- Skin Pharmacol. Appl. Skin Physiol., 2001;14:52–67dent vasoconstrictor response which was abolished by

27 27tetrodotoxin (10 M), prazoscin (10 M) and27guanethidine (10 M). In mesenteric vascular beds re- PO-100 SCHEDULE DEPENDENCY OF THE

moved from rats injected with endotoxin, the vasoconstric-CYANOGUANIDINE CHS 828 IN A RAT HOLLOW

tor responses to SNS were markedly impaired. AlthoughFIBER MODEL

genistein and L-NAME pretreatment attenuated the vascu- 1 2 1Saadia B Hassan , Lena Friberg , Elin Jonsson , Mats Olar hyporeactivity to phenylephrine, they did not modify 2 1 1Karlsson & Rolf Larsson – Division of Clinical Phar-the responses to SNS. These results indicate that genistein

macology, Uppsala University Hospital, Sweden;and L-NAME pretreatment prevent the hypotension and 2Division of Pharmacokinetics and Drug Therapy, Uppsalathe delayed hyporeactivity to phenylephrine induced by

University, Swedenendotoxin but they failed to restore the vascular hyporeac-tivity to SNS.

CHS 828 is a novel anticancer drug that has shownschedule dependent cytotoxic effects in vitro [1] as well inphase I clinical trials. The hollow fiber assay is an in vivo

PO-99 BIOMECHANICAL EVOLUTION OF IN- model for cytotoxic drug evaluation and can be used inVOLVED AND NON-INVOLVED SKIN RESULTING immunocompetent rats [2]. Our aim was to study theFROM PHOTOCHEMOTHERAPY (PUVA) OF dependence of CHS 828 effect on dosing schedule and onPSORIATIC PATIENTS time in a hollow fiber rat model. The breast cancer cell line

1 2 1˜ ´Manuel Fitas , Paulo Lamarao , Rui Minhos , Gabriela MDA-MB-231 and the leukemia cell line CCRF-CEM2 1 1˜ ´Marques Pinto , Joao Pereira & Luıs M Rodrigues – were cultured in semipermeable hollow fibers. The fibers

1Laboratory of Experimental Physiology & UCTF – were implanted subcutaneously into male Sprague Dawley´Laboratory of Cutaneous Biology, Faculdade de Farmacia rats and the rats were treated with different CHS 828 doses

da Universidade de Lisboa, Av. das Forc¸as Armadas, (125, 250, 375 and 500 mg/kg) or vehicle. The doses were21640-019 Lisboa, Portugal; Dermatology Service, Hospi- given orally either as one single dose or divided into five

ˆtal de Curry Cabral, Rua da Beneficencia 1050 Lisboa, doses given in five days. After six days the fibers werePortugal retrieved and the cell density was evaluated. Hemato-

logical toxicity and weight reduction were monitored andpharmacokinetic sampling was also performed. A five-dayThe progression of the photochemotherapy (PUVA) effica-dosing schedule produced a higher cytotoxic effect on thecy on patients suffering from different expressions ofcells, as well as tendency to more toxicity at the highestPsoriasis Vulgaris can be assessed through several bioen-dose level, compared to a single-dose schedule. The effectgineered techniques [1,2].increased in a dose dependent manner and also increasedPatients (n56) were chosen after informed consentwith increased time of exposure for both cell lines. Theseinvolving Psoriasis gutata and/or Psoriasis inplaques,.findings showed the dependence of CHS 828 effect onMethods included (a) Clinical assessment (PASI method)schedule used where a protracted schedule showed higherwith photo recording, and (b) evaluation of the skineffect compared to single dose. The pharmacokinetic and/mechanical properties (Cutometer SEM474 from CK elec-or pharmacodynamic explanations for this as well as thetronics GmbH). Measurements were performed on lesionalclinical implications need to be further explored.areas and compared with the respective negative control

[2][1] Hassan SB, Jonsson E, Larsson R and Karlsson MO.Biometrical evaluation seems to corroborate the clinical

Model for Time Dependency of Cytotoxic Effect offindings previously reported, involving a progressive re-

EUFEPS 2002 Supplement 17S (2002) S77–S85 S81

3 4CHS 828 in Vitro Suggests Two Different Mechanisms of Pharmacology, Psychiatry; Neuroscience, Universityof Action. J Pharmacol Exp Ther, 2001;299:1140–47. of California at San Diego, USA

[2] Jonsson E, Friberg LE, Karlsson MO, Hassan SB,Freijs A, Hansen K and Larsson R. Determination of 5-HT receptors are known to be critical mediators of1A

drug effect on tumour cells, host animal toxicity and central serotonergic neurotransmission, and as such, aredrug pharmacokinetics in a hollow-fibre model in rats. important potential drug targets for the treatment ofCancer Chemother Pharmacol. 2000;46:493–500. neuropsychiatric disease. Buspirone and related anxiolytics

are well-known to be partial agonists of the 5-HT1A

receptor, whereas the clinical relevance of 5-HT receptor1A

PO-101 EVALUATION OF IN VITRO CYTOTOXICITY antagonism has not yet been well defined. As part of ourOF AMPHIPHILIC b-CYCLODEXTRIN NANO- drug discovery strategy, we have systematically character-SPHERES AND NANOCAPSULES ON HUMAN FIB- ized the activity of existing drugs at most of the mono-ROBLAST AND GRANULOCYTES aminergic G-coupled protein receptors, including the 5-

1 1 2~˘ ¨Erem Memisoglu , Imran Vural , Dicle Guc¸ , Lale HT receptor using our physiologically predictive func-1A2 1 1˘Dogan , A. Atilla Hıncal – Hacettepe Univ., Fac. of tional assay (R-SAT�) [1,2]. To date, we have profiled a

2Pharmacy, Dept of Pharmaceutical Technology; Fac. of large number of clinically relevant compounds, includingMedicine, Dept of Basic Oncology, 06100 Ankara, Turkey neuropsychiatric agents. In contrast to the results obtained

with many of the other related receptor subtypes, surpris-Purpose of the study was the cytotoxic evaluation of ingly few compounds displayed potent agonist activity atnanospheres and nanocapsules prepared fromb-CDC6, an the 5-HT receptor, while a significant number of an-1A

amphiphilicb-cyclodextrin modified in the secondary face tipsychotics exhibited competitive antagonist activity atwith 6C alkyl esters [1]. Cell viability was chosen as a this site. These results shed new light on the question ofcytotoxicity parameter [2] and determined using the 3-(4,5- the clinical significance of 5-HT receptor activity.1A

dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) assay. Normal human fibroblast cells were incu- [1] Weiner DM, Burstein ES, Nash N et al., 5-hydroxy-bated in Dulbecco’s minimum essential medium (DMEM) tryptamine2A receptor inverse agonists as an-and 10% fetal bovine serum with varying concentrations of tipsychotics. JPET, 2001;299:268–276.b-CDC6 nanospheres or nanocapsules on a 96-well micro- [2] Burstein ES, Spalding TA, Brann MR. Pharmacologyplate for 72 h under 5% CO2 atmosphere at 378C. Human of muscarinic receptor subtypes constitutively acti-granulocytes were obtained by density centrifugation and vated by G proteins. Mol Pharmacol, 1997; 51:312–cytotoxicity was assessed also by cell viability determi- 319.nation by MTT assay. It was demonstrated thatb-CDC6nanospheres and nanocapsules did not cause significantcytotoxicity within the effective concentration range and PO-103 POLYAMINAMIDES-DERIVATES AS TOOLSED 50% values were reached at very high concentrations. IN DRUG RESEARCH AT THE NMDA RECEPTORThis may suggest the safety of amphiphilicb-CD WITH DIFFERENT PHARMACOLOGICAL EFFECTS

1 1 2 1 1¨nanoparticles for parenteral route as far as cytotoxicity is D Niepel , T Pohler, M L Berger , G Ecker & C R Noe1concerned. – Institute of Pharmaceutical Chemistry, University of

2Vienna, Althanstrasse 14, A-1090 Wien, Austria; Brainˆ˘[1] Memisoglu E, Bochot A, S¸en M, Charon D, Duchene Research Institute, University of Vienna, Spitalgasse4, A-

D, Hıncal AA. Amphiphilic b-cyclodextrins modified 1090 Wien, Austriaon the primary face. Synthesis, characterization andevaluation of their potential as novel excipients in the Because of its important role in a number of neurode-preparation of nanocapsules. J. Pharm. Sci., generative diseases the NMDA receptor has become a2002;91(5):1214–1224 major target in drug research. Selective ligands of the

¨[2] Muller RH, Maaben S, Weyhers H, Specht F, Lucks NMDA-receptor polyamine sites could serve as valuableJS. Cytotoxicity of magnetite-loaded polylactide, poly- tools to characterize these sites and their impact onlactide/glycolide particles and solid lipid nanoparti- receptor function and may be valuable lead structures forcles, Int. J. Pharm., 1996;138:85–94 new polyamine-type drugs.

In continuation of previous synthetical work in ourgroup a series of polyamineamides have been synthesized,

PO-102 CLINICAL PHARMACOLOGY AT THE trying to achieve significant inverse-agonistic activity withHUMAN 5-HT RECEPTOR a high selectivity. Deriving from our different thiophen-1A

1 1 1 1 1N Mohell , A Lee , E Donohue , H Berry , U Hacksell , dialkandiamines, variations of the C-8 chain have been1,2 1 1,3,4M R Brann , E S Burstein & D M Weiner – performed to study the influence of different substituents

1 2ACADIA Pharmaceuticals, San Diego, CA; Departments on receptor affinity and selectivity.

S82 EUFEPS 2002 Supplement 17S (2002) S77–S85

essential components. A potent inhibitory ditopic species,based on 2-aminobenzamidine, was identified for HPrK/P.

[1] Galinier A, Kravanja M, Engelmann R, HengstenbergW, Kilhoffer MC, Deutscher J, & Haiech J. Newprotein kinase and protein phosphatase families medi-

To study the impact of the compounds, in vitro binding ate signal transduction in bacterial catabolite repres-techniques with the radiolabeled open channel blocker sion. Proc Natl Acad Sci USA, 1998;95:1823–8.

3 ¨[ H]MK-801 have been used. All compounds tested [2] Ramstrom O, and Lehn J-M. Drug Discovery byshowed considerable affinity to the NMDA-receptor. Inter- Dynamic Combinatorial Libraries. Nat Rev Drugestingly, some compounds exhibited both agonistic and Discov, 2002;1:26–36.antagonistic effects in a concentration dependent manner.Currently, these compounds are additionally tested on ourin vitro blood–brain barrier (BBB) model to receive PO-105 REDUCTION OF LIVER DAMAGE BY ETHA-informations on their BBB permeability. NOLIC EXTRACT OF GINGER DURING COUNTRY-

We gratefully acknowledge financial support by the MADE LIQUOR-INDUCED HEPATOTOXICITY INAustrian Science Fund (project P14582-CHE) RATS

Areeg Anwer Shamsher, Uma Bhandari, K K Pillai & M SY Khan – Departments of Pharmacology and Chemistry,

PO-104 HPr KINASE/PHOSPHATASE AS A NEW Faculty of Pharmacy, Hamdard University, New Delhi-TARGET FOR POTENTIAL ANTIBIOTIC DRUGS. 1110062, IndiaIDENTIFICATION OF BIS-CATIONICHETEROCYCLIC INHIBITORS FROM A DYNAMIC Country-made liquor (CML) has been reported as a potentCOMBINATORIAL LIBRARY hepatotoxic agent [1]. In this study, we observed a

1 2 2¨ ¨H Ramstrom , T Bunyapaiboonsri , O Ramstrom , J beneficial hepatoprotective effect of Zingiber officinale1 2 1Haiech & J-M Lehn – Pharmacologie et Physico- (ginger, Zingiberaceae) against CML-induced hepato-

´Chimie des Interactions Cellulaires et Moleculaires, UMR toxicity in rats [2]. Wistar albino rats of either sex´ ´CNRS 7034, Universite Louis Pasteur, Faculte de Phar- weighing 120–200 gm were made hepatotoxic by adminis-

macie, 74 route du Rhin, B.P. 24, F-67401 Illkirch, France; tering CML (3 ml /100 gm/day in 2 divided doses) and2 ´Laboratoire de Chimie Supramoleculaire, ISIS – Uni- corn oil (1 ml /100 gm/day, in a single dose) orally for 21

´versite Louis Pasteur Strasbourg, 4 rue Blaise Pascal, days. The ethanolic extract of ginger (200 mg/kg, P.O.)F-67000 Strasbourg, France was administered from day 15 to day 21 along with CML.

Silymarin (25 mg/kg, P.O.) was used as a standardFor survival and to adapt to environmental changes in reference drug for comparing the data as its hepato-carbon sources the bacteria have developed a sophisticated protective activity is well known [3]. Serum and histo-regulatory system to sense changes in carbohydrate availa- pathological determinations were made. CML administra-bility. In low guanine, cytosine Gram-positive bacteria, tion induced marked elevations in serum levels of AST,e.g.,Bacillussubtilis, the main mechanism in the hierarchi- ALT, ALP,g-GTP, and tissue lipid peroxide levels.cal use of carbon sources seems to involve the bifunctional Histopathological examination revealed diffused and focalenzyme HPr kinase/phosphatase (HPrK/P) [1]. Since fatty changes and mild periportal fibrosis of hepatic cells.HPrK/P-deficient bacterial mutants lead to severe growth Treatment with ethanolic ginger extract significantly (P,

defects, inhibitors of this key enzyme could form a new 0.01) reversed the above levels. Histopathological exami-family of antibiotic drugs. nations of drug treatment showed almost normal hepat-

Dynamic Combinatorial Chemistry (DCC) is a new ocytes with well brought out nuclei and mild congestion inconcept in drug discovery designed to produce flexible, or the central vein.adaptive, libraries [2]. In the present study, a library was Taken together with the present results, the gingercomposed of all possible combinations resulting from the administration prevents fatty infiltration, exhibit membranedynamic interconversion of 16 hydrazides and 5 mono- stabilizing property, antioxidant activity and protects liveraldehyde or dialdehyde building blocks, resulting in li- from marked injury caused by CML. As far as we know,braries containing up to 440 different constituents. Of all these results could be the first major step for research onpossible acyl hydrazones formed, active compounds con- hepatoprotective potential of ginger in liver injury.taining two terminal cationic heterocyclic recognitiongroups separated by a spacer of appropriate structure could [1] Gulati R, Agarwal S, Agarwal SS. Hepatoprotectivebe rapidly identified using a dynamic deconvolution pro- studies on Phyllanthus embelica Linn and Quercetin.cedure. Thus, parallel testing of sub-libraries where one Indian J Expt Biol, 1995;33:261–68.specific component was excluded basically revealed all the [2] Hikino H, Kiso Y, Kato N, Hamada Y, Sankawa U et

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2al. Antihepatotoxic actions of gingerols and diaryl Armadas, 1600 Lisboa, Portugal; Plastic and Reconstruc-´hepatanoids. J Ethnopharmacol, 1985;14:31–9. tive Surgery Service, H.S. Jose, 1150 Lisboa, Portugal

[3] Ferenci P, Dragosics B, Dittrich H et al. Randomizedcontrolled trial of silymarin treatment in-patients with The complex nature of the full thickness burn injurycirrhosis of the liver. J Hepatol, 1989;9:105–13. healing process limits the knowledge of many of the

mechanisms involved. Further description of the process is,therefore, a major objective to improve therapeuticalintervention. Thus, the present study aims to characterisePO-106 ANTIAGGREGATORY ACTIVITY OF HYPO-the cutaneous lesion in allografted patients by non-invasiveGLYCAEMIC SULPHONYLUREAS

1 1 1 methodologies.Danuta Siluk , Roman Kaliszan , Piotr Haber , Jacek2 3 1 Burned patients (n57) were studied, after informedPetrusewicz & Zdzisl«aw Brzozowski – Medical Uni-

written consent. Lesion (L) and Donour (D) lesional areas˜versity of Gdansk, Department of Biopharmaceutics andwere characterised in terms of transcutaneous oxygen˜Pharmacodynamics, Gen. J. Hallera 107, 80-416 Gdansk,

2 pressure (tcpO ; TINA TCO, Radiometer, Denmark), local2˜Poland; Medical University of Gdansk, Department ofmicrocirculation (Laser-Doppler flowmetry; Oxford Op-˜Pharmacology, Do Studzienki 38, 80-277 Gdansk, Poland;

3 tronix, UK) and transepidermal water loss (TEWL,˜Medical University of Gdansk, Department of ChemicalTewameter TM210, CK electronics, FRG) and compared˜Technology of Drugs, Gen. J. Hallera 107, 80-416 Gdansk,with the respective contralateral intact areas. Measure-Polandments took place at week 4, 6, 8 and 12 weeks (W).

This variables were adequate to quantitatively describeVascular complications observed in diabetes are often the healing process, involving a progressive increase ofrelated to altered platelet functions. Abnormal platelet tcpO and a stabilisation on local microcirculation values2functions may manifest by excessive platelet aggregation.not before W12. Variable decrease in the barrier functionThe purpose of this study was determination of inhibitory (TEWL) from W4 to W6, probably reflects the progressiveeffects on platelet aggregation of hypoglycaemic sul- thickening of the scar.ponylureas and its comparison to that of acetylsalicylic Results seems to confirm that the application of theacid. An attempt to evaluate the protective effect of the above-mentioned techniques may help to identify theselected agents on pulmonary microembolism in mice and complex mechanisms involved, justifying further develop-to identify structural parameters of the agents contributing ments.to their antiaggregatory activity was also performed.

a aAntiaggregatory datain vitro was carried out for 13] ][1] Rodrigues, L.M., Almeida, M ., Mouzinho, M .,

sulphonylurea derivatives. Aggregation of platelets, incu- Quaresma, P., Ferreira, R., Serra, J. Characterizing thebated with the agents at concentrations varying from 7.5 to functional evolution of allografts in the cutaneous loss480 mmol / l, was induced by 10mmol / l ADP. In in vivo by thermal injury (port.), SOCF Proceedings, A76,pulmonary microembolism test in mice glibenclamide, 2001.gliclazide and glimepiride were used. The most pro- [2] Girbal, J.C., Gander, G. A new type of split thicknessnounced inhibition of platelet aggregation was found for skin graft, Surg Gynec & Obstet, 1966;122:326–332glimepiride, gliclazide, gliquidone, glibenclamide and [3] Graham, J., Schomacker KT, Glatter, RD, Briscoe,compound 2A. Glimepiride appeared more potent ADP- CM, Braue Jr, E., Squibb., S. Bioengineering methodsinduced platelet aggregation inhibitorin vitro than gli- employed in the study of wound healing of sulphurclazide. The microembolism test in mice confirmed anti- mustard burns, Skin Res & Technol, 2002;8:57–69platelet activity of gliclazide. Quantitative structure-activi-ty relationships indicate that antiaggregatory activity ismainly affected by electronic and not by lipophilic prop- PO-108 DOSE DEPENDENT ANTITUMOR ACTIVITYerties of the agents. The antiaggregatory activity of sul- OF LIPOSOMAL CYPHELINphonylurea derivatives might be additional to their main N Tchikineva, O Orlova, A Polozkova, I Golubeva, Itherapeutic effect, especially for glimepiride, gliclazide and Ossetrova, Z Smirnova & N Oborotova – N. Blokhingliquidone. Memorial Cancer Research Center of RAMS, Kashirskoye

sh., 24, Moscow, 115478 Russia

PO-107 FUNCTIONAL CHARACTERISATION OF THE Cyphelin (sarcolysin peptide) is an original preparationCUTANEOUS BURN HEALING PROCESS with high antitumor activity, demonstratedin vivo and in

1 1 a 2]´Luis M Rodrigues , Jose M Magro , M M Mouzinho , clinical trials. In order to increase its bioavailability we

1 1 1 a]Ricardo Ferreira , Pedro C Pinto , Nuno Furtado & M A created and studied liposomal drug formulation of

2 1Almeida – Laboratory of Cutaneous Biology, Faculdade cyphelin. Two liposomal compositions were prepared:´de Farmacia da Universidade de Lisboa, Av das Forc¸as containing lecithin and cyphelin (lip-1) and containing

S84 EUFEPS 2002 Supplement 17S (2002) S77–S85

lecithin, cholesterol and cyphelin (lip-2). Rats with sar- AV block were relatively rare. The number and types ofcoma M-1 were chosen as experimental model. arrhythmia observed are similar to those seen in previous

Both liposomal compositions demonstrated high anti- studies [1–3]. Cardiac arrhythmias are common in youngtumor effect if administered i /v in dose of 10 mg/kg healthy males although frequent ectopic beats of atrial or3-times every 72 h (TGI572–83%). This considerable ventricular origin are rarely observed. Due to difficulties ineffect remained up to the 23-th day of the experiment interpreting whether or not a drug has induced an arrhyth-(90–96% of TGI). Therapeutic activity of cyphelin sub- mia, it is advisable to exclude healthy volunteers withstance was the same (74–99% of TGI), but the total dose certain rare arrhythmias from phase 1 clinical studies. Aof the liposomal preparation only 30 mg/kg (13-times reasonable criteria would be to only use individuals withlower then the dose of substance). No any considerable no evidence of frequent VEs (.10 per hour), triplets,differences were detected between antitumor activity of salvos, VTs or SVTs.two liposomal compositions, but side effects of lip-1 weremuch more severe. So further investigations of dose- [1] Brodsky M, Wu D, Deres P, Kanakis C Jr, Rosen KM.dependent therapeutic action were conducted using lip-2. Arrhythmias documented by 24-hour continuous elec-

It was demonstrated that lip-2 in dose of 0,5 mg/kg i /p trocardiographic monitoring in 50 male medical stu-does not cause any antitumor action on M-1 (only 43% of dents without apparent heart disease. Am J CardiolTGI); at the same time dose 80 mg/kg was LD . The 1977;39:390–395.100

dose of 40 mg/kg caused the death of 2 from 8 animals, so [2] Clarke JM, Hamer J, Shelton JR, Taylor S, Venningit can be considered to be a little bit lower then MTD. The GR. The rhythm of the normal human heart. Lancetmost considerable antitumor effect was obtained in rats 1976;1(7984):508–12.given lip-2 in the dose 40 mg/kg – 98–99% of TGI by the [3] Ilson BE. Cardiovascular monitoring in normal healthy24-th day after tumor inoculation. Also 5 from 6 rats were adults: A literature review and recommendations forcured in this group (83%), the figure is statistically the reporting of disturbances of cardiac rhythm. Am Jsignificant if compared with the control (25% of animals Ther 1995;2(11):893–899.were spontaneously cured). Blank liposomes were ad-ministered in the same volumes as studied preparations,but no any antitumor or side effects were demonstrated. PO-110 AN INVESTIGATION INTO THE ANNUAL

Data obtained in this experiments will help to promote USAGE AND COST OF HORMONAL THERAPY INto the clinical trials new effective liposomal preparation for THE TREATMENT OF BREAST CANCER AT TWOanticancer treatment. BIRMINGHAM HOSPITALS IN THE UNITED KING-

DOMJ V Willis – Coventry University, B Hebron, City Hospital

PO-109 CONTINUOUS ELECTROCARDIOGRAPHIC Pharmacy, Carl Partridge, Queen Elizabeth Hospital Phar-(HOLTER) MONITORING IN 65 HEALTHY YOUNG macy, UKMALE VOLUNTEERS. AN OBSERVATION ON FIND-INGS This study investigated the annual usage of the hormonalD Wilbraham, D Amin & M Angell – Guy’s Drug treatments tamoxifen, aminogluthethimide, anastrazole,Research Unit, Quintiles Ltd. exemestane, letrozole, megestrol acetate and goserelin.

Data from the pharmacy departments of two teachingContinuous Holter monitoring for 48-hours was performed hospitals located in Birmingham that shared the same teamon 65 healthy young male volunteers aged 19–40 as part of clinical oncologists was compared for the years 1998 toof a screening procedure for a phase 1 clinical drug trial. 2000. Tamoxifen was shown to be the drug predominantlyPrior to the Holter monitoring, medical history, examina- used at both hospitals in the years 1998 and 1999. In 2000tion, laboratory blood tests and 12-lead ECG were per- this continued to be the case for one hospital but at theformed to exclude anyone with apparent cardiac disease. other the use of tamoxifen declined from 42% to 17%The results showed that many different arrhythmias whilst exemestane, used for the first time, accounted foroccurred on a frequent basis. Bradyarrhythmias and tach- 44% of the hormonal treatments used. Exemestane hasyarrhythmias were seen almost universally in the popula- been reported to be the first oral steroidal aromatasetion studied (98.5% and 95.4% respectively). 72.3% had at inactivator for the treatment of post-menopausal womenleast 1 premature normal beat observed over 48 hours, but with advanced breast cancer whose tumours fail to respondonly 2 of these volunteers had over 100 observed. Isolated to tamoxifen therapy [1] and has been shown to be costventricular ectopics (VEs) were observed in 75.4% of the effective compared with megestrol acetate [2]. Comparingpopulation studied, however frequent VEs (.100 per 48- the costs of treatment showed that the highest proportionhours or.10 per hour) were rarely seen. Other arrhyth- of the annual cost of treatment at each hospital was formias such as couplets, triplets, salvos, SVTs, bigeminy, goserelin then anastrozole and megestrol acetate. Goserelintrigeminy, and first-degree and second-degree (Mobitz 1) has become established as the drug of choice for women

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below the age of 50 years showing substantial benefits as treatment for breast cancer. Hospital Medicine,adjuvant hormone therapy for pre-menopausal breast can- 2000;61:650–655.cer patients [3]. Clearly the choice of hormonal therapy is [2] Hillner BE, Radice D. Cost-effectiveness analysis ofdependant primarily on the age of the patient and the exemestane compared with megestrol in patients withpresentation of the disease but an audit of prescribing advanced breast carcinoma. Cancer, 2001;91:484–9.patterns and their associated costs would be beneficial in [3] Early Breast Cancer Trialist’s Collaborative Group.determining a cost-benefit for the treatment of breast Tamoxifen for early breast cancer: an overview of thecancer patients. randomised trials. Lancet, 1998;351:1451–65.

[1] Johnston SRD. Irreversible aromatase inactivation: