Ehrlich II –2nd World Conference on Magic Bullets

Table 2: Percentage resistance to anti tuberculosis drugs Drug No.of resistant isolates % of resistance Isoniazid (INH) 16 12.2 Rifampin (RMP) 10 7.6 Streptomycin (SM) 13 9.9 Ethambutol (EMB) 19 14.5 Pyrazinamide (PZA) 16 12.2 p-aminosalicylic acid (PAS) 20 15.3 Ethionamide (ETA) 15 11.4 Cycloserine (CS) 15 11.4 Kanamycin (KM) 9 6.8 Ciprofloxacin (CIP) 7 5.3 Viomycin (VM) 2 1.5 Rifabutin (RIB) 1 0.7 Table 3: Antimicrobiol resistance pattern of multi-drug resistant MDR strains Anti-TB drugs Number of resistant strains INH+RMP+PZA 1 INH+RMP+CS 1 INH+RMP+PZA+KM+CS 1 INH+RMP+PZA+EMB+SM+KM+CS+PAS 1 CONCLUSIONS: 1) High rate of acquired drug resistance and the wide variation in drug sensitivity patterns was observed.2) Results in our study indicates the necessity to have an antibiotic sensitivity test before instituting treatment for recurrent TB patients.

Pharmacokinetic-Pharmacodynamic Modeling Of Aliskiren Effects On Biomarkers Of The Renin-Angiotensin System In Humans MAGER DE^1, HONG Y^1,2, DINGEMANSE J^3 ^1University at Buffalo, SUNY, Buffalo, NY, USA; ^2Bristol Myers Squibb Co., Princeton, NJ, USA; ^3Actelion Pharmaceuticals Ltd., Allschwil, Switzerland Background: Drugs targeting the renin-angiotensin system (RAS) are an important therapeutic class for the management of several cardiovascular disorders. Aliskiren is the first approved orally active renin inhibitor and exhibits a dose-dependent antihypertensive effect. The purpose of this study is to develop a semi-mechanistic pharmacokinetic-pharmacodynamic (PK-PD) to evaluate aliskiren effects on several biomarkers of RAS in humans. Methods: Mean plasma renin activity (PRA) and plasma concentrations of aliskiren, active renin (AR), angiotensin-I (ANG-I), and angiotensin-II (ANG-II) were extracted from a published 3-way crossover, placebo-controlled study. Healthy male volunteers (n=18) received either placebo or 20mg enalapril followed by 2 oral doses of aliskiren (40 and 80mg, or 160 and 640mg) once daily (6-day washout period). The final PK-PD model was fitted to the data in 3-stages: 1) multiple-dosing PK were modeled and fixed during subsequent stages, 2) single-dose PD were characterized, and 3) single- and multiple-dose PD were fitted jointly. All model parameters were estimated using the maximum likelihood method in S-ADAPT (v.1.51). Results: A two-compartment model with nonlinear elimination and distribution best described aliskiren disposition. AR increased in a dose-dependent manner following the administration of aliskiren, which was described by an indirect stimulatory response model in conjunction with an empirical sub-model of functional adaptation. In contrast to AR, PRA decreased in a dose-dependent manner and was maximally inhibited within 1 hour after aliskiren administration. This response was well captured with a direct inhibitory Emax model, and the estimated aliskiren concentration producing 50% inhibition of PRA was 0.66ng/mL, which is similar to in vitro estimates (0.33ng/mL) after correcting for plasma protein binding. Inhibition of ANG-I and ANG-II paralleled the changes in PRA, and ANG-I and ANG-II remained linearly correlated throughout the study. A reduced model was also developed excluding ANG profiles, which successfully described AR and PRA profiles after multiple-dosing. Conclusions: An integrated PK-PD model of aliskiren was developed which is consistent with the pharmacology of renin inhibition. The final and reduced models test current hypotheses of RAS inhibition by direct renin antagonism and may prove useful in the future clinical development of renin inhibitors.

Four Decades In Selegiline Research; Historical Aspects, Further Perspectives MAGYAR K Dept. of Pharmacodyn., Semmelweis Univ., Neurochem. Res. Unit., Hung. Acad. Sci. The most outstanding discovery of the Hungarian drug research still is selegiline (/-/-deprenyl), a selective irreversible inhibitor of monoamine oxidase-B (MAO-B). Our aim was to develop a potent antidepressive agent. However, because of the “cheese reaction”, appeared with other MAO inhibitors, similarly to those, it has fallen into disrepute. Selegiline survived the shock of “cheese reaction”, due to its dopamine potentiating and antioxidant capacity. It became the gold standard of MAO inhibitors possessing a wide range of pharmacological activities, many of them are not related to its MAO-B inhibitory potency. It protects the effects of neurotoxins and it combats oxidative challenge, factors leading to neurodegenerative diseases. In high concentration it has pro-apoptotic activity, while in very low concentration range (10^-9to 10^-13M) inhibits apoptosis and interferes with both the intrinsic (mitochondrial) and extrinsic (death receptor) apoptotic pathways. Metabolic conversion of selegiline results partly in propargylamine containing metabolites, most probably responsible in its antiapoptotic property, including the transcriptional and translational changes. Pharmacokinetic studies revealed that selegiline undergoes intensive first pass metabolism, so its pharmacological effects are strongly influenced by the routes of its administration. Presently, in a formula of transdermal patch, selegiline in a usual oral dose, administered to parkinsonian patients, started to be used as an antidepressive agent. Research is carried out presently to identify the metabolite, responsible for neuroprotection. Because of the concentration-dependent neuroprotective effects of selegiline an optimal dosing schedule is needed to diminish undesired adverse events, with unaltered or improved neuroprotective activity

Tetracyclines: A Historical Pitfall And Additional Concept On The Treatment Of Rickettsial Diseases MAHARA F^1, IWASAKI H^2 ^1Mahara Hospital, Tokushima, Japan; ^2Univ. of Fukui, Fukui, Japan Background: A febrile disease thought to be Scrub typhus(Tsutsugamushi disease) appeared in China in 313 A.D. The first scientific report was made by Bealtz and Kawakami in 1879. This old and historical disease was highlighted at the 13th ICID (June 2008, ML), in a symposium entitled “A neglected pathogen Orientia tsutsugamushi”. In it, the author defined the following problems. 1) This disease must be recognized as a re-emerging infectious disease. 2) Chemotherapy - Empiric treatment of rickettsioses is initiated with Doxycycline (DOXY) as the world standard, however surprisingly except for Japan. This fact suggests a great historical pitfall. In this report we present the current epidemiology of scrub typhus briefly and focus on the treatment of scrub typhus and Japanese spotted fever (JSF). Method: Epidemiologic data was collected in the MEDLINE database and the NIH Website of each country. But most of the data was collected through personal communication. The clinical data was based on our original investigations. Results: Epidemiology: Scrub typhus is known to be distributed throughout the tsutsugamushi triangle. The re-emerging increase of the cases was reported in Korea 6663 (2005), in Thailand 5094 (2001) and in Japan 957 (1984). More than 20 cases of travel acquired patients were reported worldwide. Chemotherapy: DOXY is the main antibiotic in almost all countries (Korea, Russia, China, Taiwan, Thailand, India etc.). Minocycline (MINO) is not used except for Japan. However drug resistance to DOXY has been reported. On the other hand, MINO is the standard treatment for Tsutsugamushi disease in Japan. More than 4000 cases were treated with MINO in the last ten years, and drug resistance was not reported. With a single dose of 200 mg of MINO, 90% of the patients were afebrile within 24 hrs. To clarify the mechanism, we investigated the cytokine modulation induced by tetracyclines in vivo and in vitro studies (unpub. data). Quinolones are effective for spotted fever rickettsiae though tetracyclines are more sensitive. So, the author recommended the combination treatment with MINO and Quinolone for the severe cases of JSF. We think this method is applicable for treatment of other rickettsiosis. Conclusion: 1) We proved the usefulness of MINO which is standard only in Japan among the world. 2) This fact supports the paradigm change of double bullets (DOXY and MINO), as the global standard for the treatment of Rickettsial diseases. 3) Clinicians must pay more attention to scrub typhus and other rickettsioses, and consider the double bullets to prevent travel acquired rickettsioses.

Infectious Pregnancy Complications MAHARAJ D Dept. of O&G, University of Otago, Wellington, New Zealand Epidemiological reports estimate that 7.7 million perinatal deaths occur annually worldwide, including 4.3 million that take place in late pregnancy, while the remaining neonates die in the first weeks of life. Reports attribute the majority of these consequences to infections of the fetus in utero. Infections during pregnancy affect the mother and often some infections may be transmitted to the fetus in utero, during the intrapartum period or, postnatally, with potentially serious consequences. Many infections have been linked with increased risks of premature delivery and low birth weight, and associated morbidity and mortality of both mother and child. Acute or chronic specific infectious diseases may be contracted during the course of pregnancy, and conception may occur in women already subject to an infection. The coexistence of pregnancy may aggravate the risk to maternal life in cases of the more serious of these diseases. In pregnancy most infections are no more common, nor more serious than in a non-pregnant population of women of similar age. The effects on pregnancy depend on the degree of pyrexia, its duration, and the stage of fetal development when it occurs. Mild exposures during the preimplantation period, and more severe exposures during embryonic and fetal development often result in miscarriage, premature labor, growth restriction, or stillbirth. Hyperthermia may also cause a wide range of fetal structural and functional defects, with the central nervous system (CNS) being most at risk. While there is a greater incidence of neonatal morbidity and mortality with transmitted infections, not all maternal infections lead to transmission to the fetus, nor does transmission to the fetus lead to disease or sequelae. During the puerperium, parturient women are particularly susceptible to serious infections of the genital tract and childbed fever remains one of the most important causes of maternal death. Infections in pregnancy may be viral, bacterial or protozoal, affecting both mother and fetus. Some of the infections cause fevers, while others may not; this chapter will concentrate on infections resulting in maternal pyrexia, and some other infections which may not result in maternal pyrexia, but have important implications for the pregnancy and the fetus.

Let Food Be Thy Medicine And Medicine Be Thy Food MAHGOUB SEO University of Botswana, Gaborone, Botswana Background: The relationships between food and health have been recognized centuries ago. Hippocrates quote “let food be thy medicine and medicine be thy food” dates more than 2500 years back. Early nutrition research resulted in cures for many deficiency-based diseases. Recent scientific advances have blurred the line between food and medicine, as scientists identify bioactive food components that can reduce the risk of chronic disease, improve quality of life and promote general health. Methods: Literature survey and updates on the issue of functional foods have been conducted. Results: Positive health benefits from food components not considered nutrients in the traditional definition have been identified. This has resulted in the introduction of the term “functional foods” (nutraceuticals). Functional food is any food claimed to have a health-promoting and/or disease-preventing property beyond the function of supplying nutrients. Some of the potential benefits of functional foods include reducing risk of hypertension, reduced risks of cardiovascular diseases, and benefits of antioxidants in scavenging free radicals. Functional foods can take many forms (e.g. conventional foods with bioactive components, fortified or enhanced foods specifically created to reduce disease risk). Several promising bioactive compounds in foods of pant origin including legumes, cruciferous vegetables, flaxseed, barley, soy, berries, apples, coffee, tea, and oat, have been identified and studied. Examples of functional food components currently marketed, select functional foods, key components, and potential health benefits are discussed. Some challenges facing functional foods include the need for scientific, regulatory, and business frameworks, objective evaluation of data for efficacy and safety, communication of the findings to consumers, and providing incentives to encourage R & D of these novel food products. Conclusions: Developing functional foods to improve public health requires contributions from on-going research and modifications to the current regulatory framework to facilitate the review of new functional components and their health claims. It is also imperative to communicate the correct information to the consumers.

Erythropoietin: Bold Directions And Strategies For A "Magic Bullet" Destined For Neurovascular Disease MAIESE K Division of Cellular and Molecular Cerebral Ischemia, Departments of Neurology and Anatomy & Cell Biology, Center for Molecular Medicine and Genetics, Institute of Environmental Health Sciences, Barbara Anne Karmanos Cancer Center, Wayne State University School of Medicine UHC, St. Antoine, Detroit, MI, USA Implications for the clinical utility of erythropoietin (EPO) in the nervous and cardiovascular systems continue to reach far beyond the sole treatment of systemic anemia, but the development of EPO as a safe and non-toxic therapy that does not breach borders into neoplastic growth rests heavily upon the elucidation of the cellular pathways governed by this agent. Here we discuss novel strategies for the development of EPO as a cytoprotectant in vitro and in vivo during oxidative stress. Apoptotic neuronal and vascular protection by EPO occurs within a specific pre- and post-treatment therapeutic window that requires the activation of protein kinase B (Akt1), since loss of Akt1 activity through direct pharmacological blockade of Akt1 or through the gene silencing of Akt1 expression prevents cytoprotection by EPO. Yet, intimately coupled to the robust protection by EPO are a series of unique cellular mechanisms that control STAT, ERK 1/2 proteins, wingless (Wnt), and the intracellular trafficking of the Forkhead transcription factors such as FoxO3a. For example, in a series of sequential steps, EPO maintains inhibitory phosphorylation of FoxO3a and blocks FoxO3a proteolysis to foster cell survival. Second, EPO promotes the binding of FoxO3a to 14-3-3 protein through Akt1 to sequester FoxO3a in the cytoplasm to prevent transcription of apoptotic proteins. Third, gene silencing of FoxO3a during oxidative stress significantly increases cell survival, but does not synergistically improve the cytoprotective capacity of EPO, illustrating that EPO relies upon the specific blockade of the FoxO3a pathway. Fruitful development of EPO as a "magic bullet" destined for a host of neuronal and cardiovascular disorders that avoids toxicity and maximizes therapeutic efficacy will continue to depend heavily upon the elucidation and targeting of the cellular mechanisms governed by EPO in the nervous and vascular systems.

The Critical Functions Of Biliverdin Reductase (BVR) In Insulin/Insulin Growth Factor-1 (IGF-1) And MAPK Signaling Pathways: A Potential Therapeutic Application In Treatment Of Diabetes And Cancer MAINES MD University of Rochester, Rochester, USA Background: Among eukaryotic proteins, the human(h) BVR has a more diverse and expansive spectrum of functions in the cell than any other protein. The breadth of its functions was discovered in recent years and lends itself to the development of peptide-based technology to combat diseases that are associated with disruption of normal kinase-mediated functions such as diabetes and cancer. Methods: In vitro and in cell experiments, using human embryonic kidney cells were employed to examine the role of hBVR in insulin receptor kinase/IGF-1/MAPK-regulated signaling and regulation of gene expression. Results: Data gathered identified the ability of hBVR, its variants and small fragments, in modulating cell signaling and, hence, the wide range of functions that are regulated by protein kinases. These functions include growth, differentiation, gene transcription and metabolism. Regulation of glucose uptake, induction of heme oxygenase-1 (HO-1), and cytokine and Toll-like receptor signaling were identified as potential target candidates for hBVR-based therapeutic strategies. Because hBVR blocked free radical-promoted apoptosis, regulation of its activity presents a novel drug development strategy to prolong cell survival. hBVR is nearly as effective as IGF in activating the MEK-1-ERK axis. Two eight-residue peptides, one flanking P^165 and another containing S^230 block or activate, respectively, ERK, by IGF and PKC-ζ, by TNF-α. This suggests their utility as effectors of cell cycle progression and cell differentiation. Furthermore, the peptide: KYCCSRK could specifically bind hematin, which offers a rational approach to design compounds, based on the ligand-binding property, for delivering heme or synthetic heme analogues to induce or inhibit heme-regulated gene expression, including HO-1. Conclusions: Because hBVR-based seven or eight-residue peptides can effectively modulate cell-signaling networks, they bear the promise of developing into powerful tools for inhibiting or potentiating transmission of extracellular stimuli that control gene expression and cellular functions. As additional functions of hBVR in cell signaling are uncovered, the prospect of the utility of hBVR-derived structures in therapeutic settings becomes increasingly more realistic. Disclosure Statement: This work was supported by NIH grants R01ES-04066ES and R01ES-12187. I would like to thank Philip Gibbs for help in preparation of this abstract. Based on reports in Maines MD, Phys, 2005; Lerner-Marmarosh N, et al, PNAS, 2005; Miralem T, et al, JBC, 2005; Gibbs PE and Maines MD, Int J Cancer, 2007; Lerner-Marmarosh N et al, Faseb J, 2007; Maines MD, et al, JBC, 2007; and Lerner-Marmarosh N, et al, PNAS, 2008.

Intrapericardial Cisplatin Treatment Prevents Effectively The Recurrence Of Neoplastic Pericardial Effusion Maisch B, Pankuweit S, Rupp H, Ristic A Philipps Universität Marburg, Internal Medicine–Cardiology, Germany Background: The differential diagnosis of neoplastic (NE) vs. radiation induced pericardial effusion (RE) in malignancies is the prerequisite of an adequate intrapericardial and/or systemic treatment. Methods: Out of 260 pts undergoing pericardiocentesis, 42 pts with pericardial effusion (69% m, mean age 58,8+13,2 y) were identified as NE,15 pts (66% fem, mean age 54,7+12.4 y) as RE. The aetiological assessment was highly effective because it was assisted by pericardioscopy, epi- and pericardial biopsy and pericardial cytology. Pericardial effusion and biopsy analyses included biochemistry, cytology, (immuno)histology, and PCR. Results: In NE we identified: lung cancer, 52,4%; breast cancer, 19,0%; Hodgkin’s disease, 4,8%; oesophageal cancer, 2,4%; mesothelioma, 2,4%; colon cancer, 4,8%; and undifferentiated cancer of unknown origin, 14,2%. In RE 11 pts had previous breast cancer, 4 pts bronchus carcinoma but the PE was negative for them. NE were treated with intrapericardial cisplatin (single instillation of 30 mg / m² for 24 h) in addition to the tumour-specific systemic chemotherapy. It prevented recurrence of pericardial effusion during the first 3 months of the follow-up in 92,8%, and after 6 months in 83,3% of the pts. Lung cancer patients had fewer effusion relapses at the 6 months follow-up (4,5%) than breast cancer patients (37,5%)(P<0,05). Myocardial ischemia occurred after 1/42 cisplatin instillations, but there were no other complications. RE received 500mg / m² triamcinolonacetate (Volon A) intrapericardially followed by 6 months oral treatment with colchicin (3x0,5 mg). Recurrence of effusion was prevented in 13 of 15 cases (86,6%) after 3 and 6 months. With the Touhy needle and the Marburg Attacher we reach now small effusions for intrapericardial diagnosis and treatment. Conclusions: 1) Intrapericardial treatment with cisplatin prevents recurrences of NE effectively. The treatment was more successful in lung than in breast cancer pts. 2) In RE sclerosing treatment with triamcinolonacete was equally effective. 3) Pericardioscopy adds considerably to adequate diagnosis and consecutive treatment.