Ehrlich II –2nd World Conference on Magic Bullets

Drug Potential of Nigerian MedicinE Related Plants: A SURVEY BOBOYE B AND AKHARAIYI F Department Of Microbiology, Federal University Of Technology, P. M. B. 704, Akure, Ondo State, Nigeria. Medicinal plants are plants used whole or parts to prevent and cure health problems, promote and rehabilitate nature to the living population at primary, secondary and tertiary health care deliveries. In Nigeria, the use of plants at various levels of health care delivery has been in practice for many centuries past; as old as the history of human beings. These plants belong to several families including Leguminosae, Malvaceae, Mimosaceae, Euphorbiaceae, Compositae, Acanthaceae, Cannaraceae, Passifloraceae, Rutaceae, Zingiberaceae, Bombacaceae, Olacaceae, Apocynaceae, Guttiferae, Liliaceae, Sapindaceae and Combretaceae. Specific examples of the phyto-organisms are Pericopsis, Phaulopsis, Myrobalan, Pleiocarpa, Picralima, Fig, Bhadram, Akerbia, Millefoil, Bear?s breach, Copper leaf, Acalypha, Acacia, African mallow and Baobab. sThese plants are used for different medicinal purposes such as stimulant and carminative, avert fever, coughs, asthma, flatulence, helminthic and microbial problems, cancer, hypertension, leprosy, vitamin deficiency, lack of homeostasis, pox and ulcer.

Purposeful Drug-Excipients Physico-Chemical Interactions – What does that mean for Optimization of Drug Delivery and Safety? Bogdanova S^1, Pajeva I^2, Avramova N^3, Hristova Y^2 ^1Faculty of Pharmacy, Medical University-Sofia, Bulgaria; ^2Bulgarian Academy of Sciences, Bulgaria; ^3Faculty of Chemistry, Sofia University, Bulgaria Background: Drug-excipients interactions attract research interest as a biopharmaceutical tool to influence the onset, intensity and the duration of drug performance In vivo. Aim: to improve the dissolution characteristics and optimize In vitro drug release profile of some non-steroidal anti-inflammatory drugs - indomethacin (IND), ibuprofen (IBP), naproxen (NAP)  by In vitro interactions with hydrophilic polymers - polyvinyl(pyrrolidone) (PVP), polyethyleneglycol (PEG), hydroxyethyl cellulose (HEC), sodium alginate, dextran and silica-PVP- or silica-methylmetacrylate nanohybrids. Methods: Different pharmaceutical techniques – solid dispersions-, adsorbates formation, sol-gel reactions and appropriate modern physico-chemical (FT-IR, X-ray, DSC, solid state C13 and Si29 NMR, AFM, TEM) - and in silico methods for characterization have been applied. Results: Changes in   physico-chemical properties of drugs have been achieved - polymorphic transitions (IND), inhibition of crystallization and amorphization (IND, IBP), complex formation (IND,IBP), particle size reduction. All changes are related to significant increase of In vitro drug dissolution. Special emphasis has been put upon the elucidation of the character of occurring interactions on molecular level. The modern sol-gel technique has been applied to develop Silica-PVP-IBP model nanohybrids of prolonged, pH-independent IBP release.The main advantage of this reaction is that it takes place at mild experimental conditions and enables drug immobilization into the inorganic-organic network. Conclusions: Physico-chemical transformations of some non-steroidal anti-inflammatory drugs by means of purposeful interactions with polymers and nanohybrids have been proven. Significant improvement of In vitro drug dissolution and modified drug release profiles were registered. The results give reasons to assume an increase of In vivo drug activity and safety.

Escitalopram-the first ASRI. A Magic Bullet in the treatment of depression and anxiety. BØGESØ KP Lundbeck Research DK, Ottiliavej/9, DK 2500 Valby, Denmark Selective Serotonin Uptake Inhibitors (SSRI’s) are drugs (bullets) that are targeting the serotonin transporter in order to block the reuptake of the neurotransmitter serotonin. Escitalopram has, in contrast to other SSRI drugs been shown to interact with the serotonin transporter at two different sites: the primary binding site shared with other SSRI’s and a separate allosteric binding site. A consequence of this unique allosteric interaction is a prolonged dissociation half-life from the primary binding site resulting in very efficient inhibition of the serotonin uptake. Escitalopram is therefore due to this self-enhancing effect a Magic Bullet which has been termed Allosteric Serotonin Uptake Inhibitor (ASRI). Escitalopram is the active enantiomer of the racemic drug citalopram. While the R-enantiomer originally was believed to be inactive, recent investigations have surprisingly show that the R-enantiomer inhibit the effect of the S-enantiomer. Pharmacological and clinical studies have shown a clear tendency for faster onset of antidepressant action and more than two-fold higher potency of escitalopram compared with citalopram. The presentation will focus on comparing the profile of escitalopram/ASRI with SSRI’s and discuss the mechanism behind and the consequences of the inhibition of S-citalopram by its R-enantiomer. A molecular model of the serotonin transporter will be presented as well as possible mechanisms involved in the allosteric interaction.

The influence of cytarabine and myristic acid on aspirin binding with serum albumin. Spectroscopic study BOJKO B^1, SUŁKOWSKA A^1, MACIĄŻEK-JURCZYK M^1, RÓWNICKA J^1, ZUBIK-SKUPIEŃ I^1 AND SUŁKOWSKI WW^2 ^1Department of Physical Pharmacy, Faculty of Pharmacy, Medical University of Silesia, Sosnowiec, Poland ^2Department of Environmental Chemistry and Technology, Institute of Chemistry, University of Silesia, Katowice, Poland Background: In the body drugs are transported mostly via circulatory system as the complexes with albumin. This is an important part of the drug metabolism since the bound fraction of a drug has no pharmacological effect. The simultaneous binding of other exo- and endogenous ligands (such as drugs or fatty acids) can alter protein affinity towards drug and change the concentration of its free fraction. In our study the influence of cytarabine (araC) and myristic acid (MYR) on aspirin (ASA) binding with defatted bovine serum albumin (dBSA) was investigated. Methods: NMR spectra were recorded on Bruker Avance 400 spectrometer using 5 mm tubes. For water signal suppression the presaturation method was used. All solutions were prepared in D2O. The chemical shifts of proton resonances were calculated in relation to DSS signal (0.05 ppm). Fluorescence emission spectra were recorded on Kontron SFM 25 spectrofluorimeter at excitation wavelength of 280 nm. Binding constants Ka were calculated with the use of Scatchard method. All measurements were obtained at 310K. Results: The association constants calculated for the first (subdomain IIA) and second (subdomain IIIA) class of ASA binding site is 84,2x10^3M^-1 and 1,81x10^3M^-1, respectively. ^1HNMR spectra show that dBSA and araC induce changes in chemical shifts of ASA proton resonances. The effect is more evident for the aromatic ring of ASA participating in the formation of the complex than for its methyl group. Changing of chemical shifts of ASA proton resonances are not enough significant to observe differences in drug binding in the presence of MYR. Spectrofluorimetric analysis indicate that above [MYR]/[dBSA] molar ratio 4:1 KaI and KaII of ASA-dBSA complex decrease approximately by 50 and 35%. Conclusions: 1. ASA is displaced by araC in both high and low affinity binding sites. 2. Binding of MYR with albumin influences formation of ASA complexes with albumin. This effect depends on MYR:dBSA molar ratio. 3. In case of multidrug therapy or hyperlipidemia the monitoring therapy may be necessary.

Current systemic and local therapies of intra-abdominal sepsis BONDAR VM Department of Surgery, Coffee Regional Medical Center, Medical College of Georgia, Douglas, Georgia, USA Abstract Intra-abdominal sepsis (IAS) or infection is defined as inflammation of the peritoneum due to pathogenic microorganisms and their products. IAS is not a local disease, affecting the entire body, its organs and systems, producing systemic inflammatory response syndrome (SIRS). A favorable outcome for IAS depends on the ability of host defenses to overcome pathogenic microorganisms.The current treatment of IAS consists of: 1. surgical control of the source of infection, 2. reduction of intra-abdominal bacterial inoculum and 3. supportive measures. The pharmacological and mechanical means for combating intra-abdominal bacteria and the combination of the above measures will be reviewed. The systemic and local strategies to treat primary, secondary and tertiary forms of IAS will be discussed.

A generic, multidimensional SPE-platform for undisturbed LC-MS/MS analysis of basic drugs in native biofluids BOOS K-S, MORELLO R Laboratory of BioSeparation, Institute of Clinical Chemistry, Medical Center of the University of Munich, Munich, Germany. Background: In bioanalytical LC-MS electrospray ionization is very susceptible to so called matrix effects that are caused by coeluting low (LMW) and high (HMW) molecular weight sample components. Due to the natural variation of endogenous compound concentrations in biofluids such as urine and blood plasma within an individual, between individuals and between species, matrix effects are hardly predictable. In order to minimize matrix effects we treated the native biofluids by on-line multidimensional Solid Phase Extraction (MD-SPE) prior to on-line LC-MS/MS. Methods: The MD-SPE platform (High SPEed, CTC Analytics) relies on the combination of two SPE columns. The first SPE column (25 x 2 mm ID) is packed with Restricted Access Material (RAM; LiChrospher ADS RP 4, dp 25 µm, Merck KGaA) which allows size-exclusion (SEC: 1st dimension) and reversed phase chromatography (RPC: 2nd dimension). The second SPE column (20 x 2.1 mm ID) contains a mixed-mode phase (MMP; Oasis WCX, dp 30 µm, Waters) which allows weak cation exchange (IEX: 3rd dimension) and hydrophobic interaction chromatography (HIC: 4th dimension). Analytical separation of the model analytes (antidepressants) was achieved on a Zorbax RX C18 column (75 x 4.6 mm ID, dp 3.5 µm, Agilent). A Quattro Micro MS (Waters) was used in ESI(+) MRM mode. Matrix effects were monitored in post-column infusion experiments. Results: The RAM-SPE column repeatedly and very efficiently depleted plasma samples (20 µL) with regard to HMW matrix components such as proteins and quantitatively extracted the target analytes (pka value higher than 6.5) in less than 60 sec. After transfer onto the MMP-SPE column residual LMW matrix components (< 15 KDa) were eluted to waste within 60 sec. This extensive and highly selective sample clean-up resulted in almost no detectable matrix effects. In addition, MS-scans (m/z range from 100 to 1000) of blank plasma and urine from different individuals and species revealed no masses, i.e. no residual matrix components [1]. Conclusion: A combination of orthogonal chromatographic separation modes, i.e. SEC-RPC-IEX-HIC, for on-line SPE of native biofluids such as plasma and urine completely eliminates matrix effects in LC-ESI-MS/MS detection of basic drugs. [1] K.Georgi, K-S Boos, Chromatographia 63 (2006) 523-531

Control of ceramide levels by ceramide kinase: evidence from knockout animals and use of a novel potent inhibitor GRAF C, BOATH A, ROVINA P, SCHANZER A, NIKOLAY R, REUSCHEL, R, NIWA, S, BORNANCIN F. Novartis Institutes for BioMedical Research, Brunnerstrasse 59, A-1235 Vienna (Austria) Background: The sphingolipid ceramide is an important regulator of cell biology (e.g. apoptosis, differentiation) and its levels therefore must be tightly controlled. Ceramide kinase (CerK) is a unique enzyme that specifically phosphorylates ceramide into the bioactive lipid ceramide-1-phosphate (C1P). Recent data from our laboratory have provided compelling evidence that CerK is a key regulator of both C1P and ceramide levels. Methods: CerK-deficient (Cerk^–^/^–) Balb/C mice were generated as described in Graf et al. J Immunol 2008, 180:3457-66, and compared to control litermates in all studies. The diamino-benzothiazole derivative NVP-231 was synthetized as decribed in Graf et al. Mol Pharmacol 2008 74(4), in press. CerK activity assays using either fluorescently labeled ceramides or ^32P-ATP were described in Boath et al, J Biol Chem 2008 283:8517-26 and Rovina et al, Biochem J 2006 400:255-65, respectively. Growth and culture of COS-1 cells and bone marrow derived macrophages as well as Liquid Chromatography/Mass Spectrometry (LCMS) analysis was performed as decribed in Graf et al. J Immunol 2008, 180:3457-66. Results: First, using a fluorescently labeled ceramide molecule to trace CerK activity we found that C1P is short-lived compared to other ceramide metabolites, thus fulfilling an essential criteria for signaling function. This suggested that CerK activity might be useful to dispose of excess ceramide. Then we profiled Cerk^–^/^– mice and we found indeed that ablation of CerK not only decreases C1P levels but also leads to an increase in ceramide levels. Finally, we identified and characterized NVP-231, the first potent, specific and reversible CerK inhibitor. Consistent with the observations in Cerk^–^/^– mice, preliminary experiments with NVP-231 suggest that CerK inhibition may have potential to regulate C1P and ceramide levels. Conclusions: These results establish CerK as a novel, key regulator of ceramide levels and support further exploration of CerK inhibition as a rationale for the treatment of proliferative disorders.

Discovery of novel non-cyclam polynitrogenated CXCR4 coreceptor inhibitors PETTERSSON S^1, PÉREZ-NUENO V^1, ROS-BLANCO L^1, CLOTET B^2, CLOTET-CODINA I^2, ARMAND-UGÓN M^2, ESTÉ J^2, TEIXIDÓ J^1, BORRELL JI^1 ^1IQS, Universitat Ramon Llull, Barcelona, Spain; ^2Retrovirology Laboratory IrsiCaixa, Badalona, Spain. Background: CXCR4 and CCR5, chemokine coreceptors of the primary receptor (CD4) for the HIV cell fusion and entry, have been validated as targets for therapeutic intervention against AIDS. Bicyclams were the first non peptidic low molecular weight compounds with specific interaction with CXCR4, the most potent bicyclam being AMD3100 (IC50 = 1-10 ng/mL). However, it showed poor oral absorption and toxicity related to its high positive charge at physiological pH. To overcome such problem, we designed a combinatorial library of tetraamines 1, which preserve the main features of AMD3100: a) at least two nitrogen atoms on each side of the p-phenylene moiety, one in the benzylic position and the other(s) in a heterocyclic system and b) similar distances between such nitrogen atoms with those present in cyclam. Methods: 19 compounds were initially selected by evaluating a series of molecular 2D and 3D descriptors. A PCA reduced the initial set of descriptors to 5 components which were used for the diversity selection. Anti-HIV activity (EC50) and cytotoxicity (CC50) measurements were carried out in MT-4 cells infected with HIV-1. Results: The first subset of compounds showed EC50 in the range 0.9-18 g/mL. A second subset of 17 compounds afforded 12 compounds presenting EC50 in the range 0.2-2.7 g/mL. The third and final subset, covering up the total of 53 synthesized, was selected using QSAR techniques and ligand and structure-based drug design (using our CXCR4 and CCR5 modeled coreceptors). Among them, 1{8,8} showed an EC50 value of 0.008 g/ mL and a CC50 > 25 g/mL, presenting nearly the same activity as AMD3100 but showing no cell toxicity at tested concentrations. Conclusions: 1) A diversity oriented selection has allowed the synthesis of tetraamines 1 covering a broad range of activity values, useful for QSAR calculations. 2) This approach afforded compound 1{8,8}, with an EC50 value of 0.008 g/mL and a CC50 > 25 g/mL. 3) Studies on the mode of action of compounds 1 showed specific inhibition of the CXCR4 coreceptor. References - S. Pettersson, V. Pérez-Nueno, L. Ros-Blanco, R. Puig de La Bellacasa, M. O. Rabal, X. Batllori, B. Clotet, I. Clotet-Codina, M. Armand-Ugón, J. Esté, J. I. Borrell, J. Teixidó, Chem. Med. Chem. 2008, DOI 10.1002/cmdc.200800145 (July 31) - S. Pettersson, L. Ros-Blanco, J. Teixidó, X. Batllori, R. Puig de La Bellacasa, J. I. Borrell, S. Nonell, M. O. Rabal, V. Pérez-Nueno, J. Esté, I. Clotet-Codina, M. Armand-Ugón, WO 2008/049950 (02.05.2008)

Heparin as an inhibitor of cancer progression; the role of selectins BORSIG L^1, VARKI A^2 ^1Univ. of Zürich, Zürich, Switzerland; ^2Univ. of California, San Diego, U.S.A. Background: Heparin and low molecular weight heparin (LMWH) are widely used in cancer patients at risk of venous thromboembolism, which is a recognized complication of malignant disease. Recent clinical trials with LMWH and meta-analysis of earlier clinical trials with unfractionated heparin indicate that heparin affects also cancer progression. Meanwhile, heparin and LMWH were repeatedly shown to reduce metastasis in a variety of animal models. Heparin is a natural product, comprising a polydisperse mixture of highly sulfated glycosaminoglycan chains, only a fraction of which bind antithrombin. Methods: Experimental metastasis study was performed in wild type as well as in P- and/or L-selectin deficient mice. Treatment with heparin or their derivatives was performed only shortly before or after the application of tumor cells. The extend of metastasis was evaluted after 28 days. Results: We provided evidence that heparin has an additional biological activity which inhibits binding of P- and L-selectin to their natural ligands. Since also modified heparins without any anticoagulant activity were found to attenuate efficiently metastasis, the heparin effect on cancer progression may not be principally due to inhibition of coagulation. In fact, we have demonstrated that the selectins play critical roles during the hematogeneous phase of carcinoma metastasis in animal models. Natural selectin ligands on carcinoma cells were identified as mucins carrying sialylated, fucosylated and sulfated carbohydrate structures. Heparin treatment in wild type mice resulted in attenuation of metastasis similar to the one observed either P- and/or L-selectin in mice, indicating that inhibition of selectins significantly contributes to the anti-cancer effect. Inhibition of selectin-mediated interactions with carcinoma cells was found to be critical for early stages of a metastatic cascade. Most importantly, attenuation of metastasis can be achieved at clinically acceptable dosages. In addition, heparin contains also other biological activities including inhibition of heparanase, modulation of chemokines and growth factors activities. Conclusions: The available evidence from preclinical analyses, together with the promising observations from clinical trials, merits further investigation of heparin as a potential anti-metastatic therapy.

According to registration: BASU Chinmay Alterations in Female Reproductive Organs of Cyclic Rats Treated with Aqueous Extract of Moringa Oleifera Lam: Indication of Possible Role in Epithelial Ovarian Cancer BOSE CK Cancer Clinic, Cancer Nirnaya O Chikitsa Kendra, Kolkata, India. Background: A hormonal etiology of epithelial ovarian cancer has long been suspected seeing its incidence menopausal age, and now the role of FSHR has also been demonstrated. Many ovarian cancer cell line express FSHR in them. In studies of the anticancer potential of plants used in folk medicine of Bengal, extracts of plants such as Oroxylum indicum, Moringa oleifera lam, Aegles marmelos could be considered as potential sources of anticancer compounds. Amongst them only Moringa oleifera lam has unique anticancer as well as hormonal property, which may or may not be attributable to isothiocyanate or glucosinolate that it contains. An animal experiment was planned to see effect of Moringa root extract in female reproductive system of mice. Methods: 5 adult female mice of swiss strain of 30 gm each 2-control, 3-treated kept on stock diet, pellet, having nutritional value of 7 daysAn aqueous extract of the root was prepared according to a traditional method. In brief, 500 g of the root were placed in a container with 750 ml of water and boiled for 30 min. The preparation was left standing to cool and was then filtered. The filtrate, containing 66.7 mg root in 1 mL, was placed in small vials and kept in 4 degree C refrigerator until use. 1 ml of extract was used orally daily for 45 days. Results: Attenuation of ovary and uterus was seen while mice tolerated the herb extrct well. There was reversal to pre estrus phase of adult mice as was revealed by PAP smear from vagina. In histology there was absence of follicles in comparison to control ovary. There was lesser amount of fibrosis in treated ovary. Conclusion: Isothiocyanate of Moringa may inhibit proliferation of ovarian granulosa and other cells as it induces apoptosis via caspase-9 and -3 pathways, a family of calcium-dependent cysteine proteases. It may also act by inhibiting ERK1/2 and Akt survival signaling while simultaneously activating pro-apoptotic p38 and JNK1/2. There are reports to show that Moringa induced decrease in cerebral dopamine and norepinephrine which may influence and lower NGF and FSHR through central mechanisms. There is strong possibilty of using this agent in epithelial ovarian cancer and, as such, a cell line experiment is urgently necessary.

Serum and Alveolar Concentrations of Antibiotics during the Treatment of Ventilator-Associated Pneumonia BOSELLI E^1,2, BREILH D^3,4, ALLAOUCHICHE B^1,2 ^1Hosp. Édouard Herriot, Lyon, France; ^2Univ. of Lyon, Lyon, France;^3Hosp. Haut-Lévêque, Pessac, France, ^4Univ. of Bordeaux II, Bordeaux, France. Background: We assessed the serum and alveolar concentrations of antibiotics frequently used during the treatment of ventilator-associated pneumonia (VAP) in critically ill patients in order to optimize antimicrobial treatment in this particular population. Methods: Various antibiotics (β-lactams, tobramycin, levofloxacin or linezolid) were administered in intermittent or continuous infusion to critically ill patients with VAP. At steady-state (after two days of therapy), blood samples were withdrawn from each patient and serum concentrations were measured by high-performance liquid chromatography. Simultaneously to blood sampling, antibiotic concentrations were determined in epithelial lining fluid (ELF) obtained from standardized mini-bronchoalveolar lavage, which is a reliable method for the measurement of alveolar drug concentrations. Results: The 122 patients were of similar age (50-70 yrs), weight (60-80 kg) and creatinine clearance (60-100 mL/min). The antibiotic daily dose for each patient, administered in intermittent or continuous infusion, the serum and ELF concentrations and the ELF/serum concentration ratios appear in Table 1. Conclusions: Serum and alveolar concentrations and alveolar diffusion of antibiotics from both different and similar classes exhibit wide variations in critically ill patients with VAP. Individual antibiotic dosages may be helpful to optimize the administration regimen of antibiotics and therefore PK/PD parameters (T>MIC, Cmax/MIC and AUC/MIC) during the treatment of VAP. Table 1. Mean steady-state serum and ELF antibiotic concentrations

Therapeutic effects of Crocus sativus (saffron) on respiratory diseases BOSKABADY MH, ASLANI MR, AHMADZADEH H, GHASEMZADEH RAHBARDAR M, NEMATI A Dept. of Physiology and Pharmacological Research Centre, Medical School, Mashhad University of Medical Sciences, Mashhad, Iran As indicated in ancient Iranian medical books Crocus sativus (Iridaceae) or saffron has therapeutic effects on respiratory diseases. Therefore in a series of experiments, the follwing effects of cumulative concentrations of saffron and its constituent, safranal was examined on tracheal chains of guinea pigs: relaxant, stimulatory effect on -adrenoceptors, inhibitory effect on histamine (H1) receptors and the inhibitory effects on muscarinic receptors In precontracted trachea by 10 µM methacholine (roup 1) all concentrations of theophylline, extract and safranal showed significant relaxant effects compared to that of saline (p<0.05 to p<0.001). In precontracted trachea by 60 mM KCl (group 2) also theophylline, extract and safranal showed concentration dependent relaxant effects compared to that of saline (p<0.05 to p<0.001 for different concentrations except two low concentrations of safranal). However, in group 3 (precontracted incubated tissues with propranolol, chlorpheniramine and atropine by 60 mM KCl) the extracts of Crocus sativus showed a weak relaxant effect (p<0.05 only for highest concentration). There were clear leftward shifts in isoprenaline curves obtained in the presence of only higher concentration of the extract in group 1(non incubated tissues) and it’s both concentrations in group 2 (incubated tissues with chlorpheniramine) compared with that of saline. The EC50 (the effective concentration of isoprenaline, causing 50% of maximum response) obtained in the presence of both concentrations of the extract and safranal in group 1 and only in the presence of two concentrations of the extract in group 2 was significantly lower compared to saline (p<0.05 to p<0.001). The maximum responses obtained in the presence of both concentrations of the extract and safranal in group 1 were significantly lower than that of saline (p<0.005 for all cases). EC50 histamine obtained in the presence of chlorpheniramine, all concentrations of the extract and safranal in all three groups (incubated trachea with: 1) indomethacin, 2) indomethacin, propranolol, and atropine and 3) indomethacin and propranolol) were significantly greater than those of saline (p<0.05 to p<0.001) except medium concentration of the extract in group 2 and its low concentration in group 3 (0.5 and 0.25 mg/mL respectively). The EC50 obtained in the presence of all concentrations of extract and safranal in group 2 were greater than group 1 and 3 (p<0.05 to p<0.001). Maximum response obtained in the presence of all concentrations of extract and safranal in group 2 were greater than those of group 1 and group 3 (p<0.05 to p<0.001). The results of Inhibitory effects on muscarinic receptors showed clear parallel rightward shifts in methacholine-response curves obtained in the presence of atropine, 2 low concentrations of safranal and extract compared with the curves obtained in the presence of saline. The EC50 obtained in the presence of atropine, two lower concentrations of safranal and all concentrations of the extract was significantly higher than that of saline (p<0.01 to p<0.001). The maximum responses obtained in the presence of all concentrations of the extract was significantly lower than that of saline (p<0.01 to p<0.001). These results showed a potent relaxant effect of saffron, a relatively potent stimulatory effect of the extract from Crocus sativus on ß 2-adrenoceptors, an inhibitory effect at histamine H1 receptors and a possible inhibitory effect at muscarinicreceptors on tracheal chains of guinea pigs. The results also indicated that the safranal is, at least in part, responsible for the relaxant effect of Crocus sativus.

Different therapeutic effects of Nigella sativa BOSKABADY MH, MOHSENPOOR N, TAKALOO L, JAVAN H, FARHADI J Dept. of Physiology, Medical School, Azadi Square, Mashhad University of Medical Sciences, Mashhad, IR Iran In a series of studies the following effects of Nigella sativa on guinea pig tracheal chains and human airways were studied. The results showed: (1) Aqueous and macerated extracts from Nigella sativa showed significant relaxant effect compared to saline (p<0.001). The extracts caused a non parallel rightward shift in methacholine concentration response curve and EC50 methacholine obtained in the presence of extracts were significantly greater than that of saline (p<0.05 to p<0.001). (2) plant extracts caused parallel right ward shifts in histamine concentration response curves obtained compared to saline. The EC50 histamine in the presence of extracts were significantly greater than slain (p<0.05 to p<0.001). (3) There was a leftward shift in isoprenaline concentration response curve in the presence of aqueous extract. (4) Different extracts showed significant relaxant effect on tracheal chains incubated with Chlorpheniramine and propranolol, contracted by methacholine (p<0.01 to p<0.001) but did not show any relaxant effect on tracheal contracted by KCl. (5) Aqueous extract caused a rightward shift in the CaCl2 response curves and EC50 CaCl2 hin the presence of axtract was significantly greater than the presence of saline (p<0.05). (6) However, thymoquinon, main constituent of the plant did not show any significant relaxant effect on tracheal chains. (7) Boiled extract of this plant caused significant increases in all measured pulmonary function tests (PFTs), (p<0.05-p<0.001) comparable to the effect of thophylline. (8) Two months administration of boiled extract also caused significant improvement in PFT values, respiratory symptoms, chest wheeze and drug usage in asthmatic patients. (9) Concentrations of extracts of the plant showed significant reduction of cough number (p<0.001 for all cases) which were significantly greater than that of codeine (p<0.05-P<.001). Results showed a potent relaxant effect of Nigella sativa on tracheal chains, a relatively potent bronchodilatory effect on asthmatic airways and a potent antitussive effect.

Rediscovering antibiotics of alternative medicine: case of apitherapy BOUKRAA L; HAMMOUDI SM; BENBAREK H; BENHANIFIA MB; AISSAT S; AHMED M; MESLEM A Laboratory of Microbiology, Department of Veterinary Sciences, Tiaret University, Algeria Background: Antibiotic-resistant bacteria continue to be of major health concern world-wide. Since the use of antibiotics became widespread, bacteria have progressively developed resistance. Consequently, scientific efforts have been made to develop new compounds to be used beyond conventional antibiotic therapy. It has been proposed that the healing effect of bee products could be due to various physical and chemical properties. Hydrogen peroxide, volatiles, organic acids, flavonoids, beeswax, nectar, pollen and propolis are, among others, the chemical factors that provide antibacterial properties. Methods: Bacteria of medical concern such as Pseudomonas aeruginosa Escherichia coli and Methicillin Resistant Staphylococcus aureus were subjected to the action of bee products; namely honey, propolis, royal jelly and bee venom. In the majority of cases, the minimum inhibitory concentrations (MICs) were obtained using the incorporation method. The MIC was determined by finding the plates with the lowest concentration of honey on which the strain would not grow. Results: all bee products have shown antibacterial activity. Sensitivity of bacteria to bee products varies considerably within the product and the varieties of the same product. Propolis has been found to have the strongest action against bacteria. This is probably due to its richness in flavonoids. Antibiotic-resistant bacteria have been found to be sensitive to the action of hive products. Conclusion: The frequent use of antibiotics has lead to the emergence of antimicrobial resistance. Honey and other bee products were subjected to laboratory and clinical investigations during the past few decades and the most remarkable discovery was their antibacterial activity. The emergence of antibiotic resistant bacteria, particularly methicillin-resistant Staphylococcus aureus (MRSA) has posed problems in the management of chronic wound infections. Many studies have shown that application of hive products to severely infected cutaneous wounds is capable of clearing infection from the wound and improving healing. Bee products are natural products; without adverse effects on tissues, they can be safely used on burns and inserted into cavities and sinuses to clear infection.

Neurotensin Agonists: Novel Analgesics with Synergy to Morphine BOULES M^1, SHAW A^1, BARBUT D^2, RICHELSON E^1 ^1Mayo Clinic, Jacksonville, FL, USA ; ^2Sarentis Therapeutics, New York City, NY, USA Background: Neurotensin (NT) is a widely distributed neuropeptide in the central nervous system that modulates nociception at several different levels, but is associated with hypotension and hypothermia.  NT exerts its effect through NT receptors, of which there are three known subtypes (NTS1, NTS2 and NTS3).  Morphine is a µ-opioid receptor agonist that is commonly used for the treatment of many types of pain, but it is usually associated with side effects that can be serious. We hypothesize that selective NT receptor agonists may represent a novel class of analgesics and their use in conjunction with morphine will have synergistic properties with morphine which may reduce the dose administered and its side effects. Methods: Studies were done to test for the use of a novel peptide analog of NT (NT69L) as a new class of analgesic drugs, acting through the NT receptors, alone and in combination with morphine. The antinociceptive activity of NT69L and morphine was studied in rats using the hot plate test to determine if there is synergism between the two drugs in reducing pain.  The NTS2 receptor antagonist, levocabastine was used to determine the receptor subtype involved in the synergistic effect of NT69L and morphine. Results: The administration of both NT69L and morphine resulted in a dose-dependent analgesic effect.  Isobolographic analysis was used to study the antinociceptive interactions between the two drugs. The isobolographic analysis demonstrated that the combination of sub-analgesic doses of NT69L and morphine was synergistic in the hot plate test.  Pretreatment with the NTS2 receptor antagonist, levocabastine, attenuated the synergistic effect of NT69L and morphine in the hot plate test.  Conclusion: The results provide preliminary data supporting the hypothesis that the synergistic combination of NT69L and morphine would improve the pharmacological treatment of pain while minimizing specific adverse effects of each of the drugs at a higher dose.  Both NT receptors NTS1 and NTS2 are important for the synergisitic effect of NT69L and morphine.

Current Medical Countermeasures (Vaccines- Antibodies- Antibiotics) to Protect Humans from the Anthrax Bioterrorism. BOUZIANAS DG Technological Educational Institute of Thessaloniki, Thessaloniki-Macedonia, Greece. Background: B. anthracis spore attacks through the US mail system have demonstrated their feasibility as a bioterrorism weapon. Vaccination appears to be the most effective and economical form of mass protection, however, the current vaccines have drawbacks that justify the immense efforts for the development of improved treatment modalities. This review summarizes the current human approaches developed mainly since the 2001 events against inhalational anthrax. Methods – Results: The increased research activity has led to a huge expansion in the existing literature. The present work is based on an extensive review of the international literature. Combination of postattack prophylactic vaccination with antibiotic therapy is the most effective strategy. Current approaches against anthrax toxins are focused on agents that affect crucial steps of the intoxication process. High-affinity toxin-specific monoclonal antibodies have a significant clinical effect, if they are administrated rapidly. While effective antibiotics, antitoxins and vaccines are available, concerns over their safety and effectiveness have driven the development of 2nd and 3rd generation products that act rapidly and with minimal adverse effects. Protective antigen (PA) is the principal immunogen of the 1st generation licensed vaccines. A 2nd generation vaccine is based on highly purified recombinant PA (rPA) and is likely to receive licensing approval. The 3rd generation vaccines aim to enhance the efficacy of the previous vaccines. They would ideally be given via the oral, nasal or dermal routes for delivery of rPA in a single dose facilitating stockpiling and mass vaccination programs. DNA vaccination could form the basis for multiagent vaccine development. The development of novel agents is hampered by the difficulty in demonstrating effectiveness in humans. Conclusions: Treatment response to a deliberate release of B. anthracis spores includes the prompt administration of antibiotics. There are concerns for the available antitoxins and vaccines over their effectiveness and toxicity. Despite the intensive anthrax research, there has been as yet no real progress. The huge efforts are expected to provide an array of novel protective agents and their balloning list reflects the emergency of the global community to combat the anthax threat.