20^th International Conference on Pharmaceutical Biotechnology December 07-09, 2017 Madrid, Spain

Byung Woo Kim has completed his PhD in Pharmacology from Busan University, Busan, Republic of Korea. He is currently working as a Professor at Division of Applied Bioengineering, Biopharmaceutical Engineering Major, Dong-Eui University and as the Director of Blue-Bio Industry Regional Innovation Center, Dong-Eui University, Busan, Korea. His research field is Pharmaceutical biotechnology of natural Products.

Osmanthus matsumuranus, a species of Oleaceae, is found in East Asia and Southeast Asia. The bioactivities of O. matsumuranus have not yet been fully understood. Here, we studied on the molecular mechanisms underlying anticancer effect of ethanol extract of O. matsumuranus (EEOM). EEOM showed the cytotoxic activities in a dose-dependent manner in various cancer cell lines, but not in normal cells, and HepG2 cells were most susceptible to EEOM-induced cytotoxicity. EEOM induced G2/M arrest in HepG2 cells associated with decreased expression of cyclin-dependent kinase 1 (CDK1), cyclin A and cylcin B, and increased expression of phospho-checkpoint kinase 2, p53 and CDK inhibitor p21. Immunofluorescence staining showed that EEOM-treated HepG2 increased doublet nuclei and condensed actin, resulting in cell rounding. Furthermore, EEOM-mediated apoptosis was determined by Annexin V staining, chromatin condensation and DNA fragmentation. EEOM caused upregulation of FAS and Bax, activation of caspase-3, -8, -9, and fragmentation of poly ADP ribose polymerase. These results suggest that EEOM efficiently inhibits proliferation of HepG2 cells by inducing both G2/M arrest and apoptosis via intrinsic and extrinsic pathways, and EEOM may be a possible candidate for the anticancer drug development.

Soojung Jin has completed her PhD in Immunology from Osaka University, Osaka, Japan. She is presently working as an Assistant Professor at Blue-Bio Industry Regional Innovation Center, Dong-Eui University, Busan, Republic of Korea. Her research is focused on bioactive natural products and oncology.

Machaerium cuspidatum, a canopy liana, is a species of genus legume in the Fabaceae family and contributes to the total species richness in tropical rain forests. In the present study, we investigated the anti-oxidative and anti-cancer effects of M. cuspidatum and the molecular mechanisms of its anti-cancer activity in human lung adenocarcinoma A549 cells and human hepatocellular carcinoma HepG2 cells. Methanol extract of M. cuspidatum (MEMC) showed significant anti-oxidative activity and the cytotoxic effect in a dose-dependent manner in several cancer cell lines. Annexin V-positive apoptotic cells and apoptotic bodies increased by MEMC treatment. Further investigation showed that MEMC-induced apoptosis was associated with the increase of p53 and Bax expression, and the decrease of Bcl-2 expression. In addition, MEMC treatment led to proteolytic activation of caspase-3, -8, -9 and degradation of poly ADP ribose polymerase (PARP). Taken together, these results suggest that MEMC may exert a beneficial anti-cancer effect by apoptosis induction via both extrinsic and intrinsic pathways in A549 and HepG2 cells.

Hyun Ju Kwon has completed her PhD in Bioengineering from Osaka University, Osaka, Japan. She is presently working as a Professor at Division of Applied Bioengineering, Biopharmaceutical Engineering Major, Dong-Eui University and as an Assistant Director of Blue-Bio Industry Regional Innovation Center, Dong-Eui University, Busan, Korea. Her research field is Natural Products and Oncology.

Julbernardia globiflora, a tropical African tree widespread in Miombo woodland, has been used in folk medicine for the treatment of depression and stomach problems. However, the bioactivities of J. globiflora have not yet been fully determined. The objective of this study was to evaluate the antioxidative and anticancer effects of methanol extract of J. globiflora (MEJG) and the molecular mechanism of its anticancer activity in human colon carcinoma HT29 cells. MEJG exhibited significant antioxidative activity and cell growth inhibitory effect on HT29 cells in a dose-dependent manner. MEJG induced apoptosis of HT29 cells with the increase of apoptotic cells and apoptotic bodies using Annexin V staining and 4,6-diamidino-2-phenylindole (DAPI) staining, respectively. The MEJG-induced apoptosis was associated with the increase of Fas, a death receptor, and Bax, a pro-apoptotic protein, and the decrease of Bcl-2, an anti-apoptotic protein, resulting in the release of cytochrome c from the mitochondria into the cytosol and activation of caspase-3, -8 and -9. The apoptotic effects of MEJG were confirmed by cleavage of poly (ADP-ribose) polymerase (PARP). Collectively, these results suggest that MEJG may exert the anticancer effect in HT29 cells by inducing apoptosis via both the intrinsic and extrinsic pathways.

You Na Oh has completed her Master in Microbiology from Dong-Eui University, Busan, Republic of Korea. She is currently working as a researcher at Blue-Bio Industry Regional Innovation Center, Dong-Eui University. Her research is focused on Bioactive Natural

Sorbus rufopilosa, a tsema rowan, is a species of the small ornamental trees in the genus Sorbus and the family Rosaceae found in East Asia. The antioxidant and anticancer effects of S. rufopilosa remain unclear. The objective of this study is to evaluate the antioxidant and anticancer effects of ethanol extract of S. rufopilosa (EESR) and the molecular mechanism of its anticancer activity in human colon carcinoma HT29 cells. EESR showed significant antioxidant activity and inhibitory effect on HT29 cell growth in a dose-dependent manner. EESR induced cell cycle arrest at G2/M phase in a dose-dependent manner by modulating the expression of cyclin B, cyclin-dependent kinase 1 (CDK1), and CDK inhibitor p21. EESR-induced apoptosis was associated with the upregulation of p53, a death receptor Fas, a pro-apoptotic protein Bax and the activation of caspase 3, 8, and 9, resulting in the degradation of poly ADP ribose polymerase (PARP). These results suggest that EESR efficiently inhibits proliferation of HT29 by inducing both cell cycle arrest and apoptosis, and may be a possible candidate for the anticancer drug development.

Biofilm-associated wound infections have been considered a life-threatening infection that affects millions of people each year and are among the major cause of infectious disease-related mortality and morbidity worldwide. Bacterial biofilms protect bacteria from host immune responses and promote strong resistance to antibiotic treatment which leads to impaired wound healing, hospitalization and amputation particularly in chronic wound such as diabetic foot ulcer. Recently, nitric oxide (NO) has emerged as novel agent in biofilm dispersal and accelerates wound healing. In this study, we investigated the potency of positively charge NO-releasing PLGA/PEI nanoparticles (NO/PPNPs) for adhesion on biofilm surface that elevate biofilm dispersal and wound healing efficacy. Poly (lactic-co-glycolic acid) (PLGA) were used to incorporate polyethyleneimine (PEI)/NO adduct (PEI/NONOate) by an oil-in-water (O/W) emulsion evaporation method to form NO/PPNPs. Adhesion of NO/PPNPs on bacterial biofilm and the progress of in vivo biofilm dispersal were performed in biofilm wound and characterized by 3D confocal microscopy. In vivo biofilm was prepared by inoculating Methicillin-Resistant Staphylococcus aureus (MRSA) suspension on the surface of wound in db/db mouse (type-2 diabetic). Photographs of the wounds were taken to observe the gross visual wound healing. Furthermore, histological analysis was performed with H&E and Masson trichrome stain to observe the skin morphological and collagen deposition, respectively. Positively charged of NO/PPNPs facilitated the electrostatic binding to the negatively charged biofilm matrix, thereby increasing the biofilm dispersal by NO released from NO/PPNPs. NO/PPNPs treatment a biofilm-challenged diabetic mouse accelerated wound healing as compared to untreated and blank nanoparticles. In addition, histological examination revealed that wounds treated with NO/PPNPs showed increased numbers of fibroblast-like and collagen deposition with healed skin structures close to the normal healthy epidermis. Thus, the NO-releasing polymeric nanoparticles investigated in this study could be a promising approach for the treatment of biofilm-challenged chronic wounds and various skin infections.

Ana Despina Ionescu has her expertise as the Director of different Romanian Research Projects related to the bioremediation of the industrial waste waters, the use of some biological filtering systems in order to reduce the toxic contaminants of some natural water resources and the establishment of requested parameters in order to use the natural mineral water springs for the public health improvement. She is a PhD in the field of Industrial Biology and she has obtained more Medals at the International Innovation Fairs from Brussels and Geneva during the period 2005-2014. She is working as a Senior Scientific Researcher at the National Chemical-Pharmaceutical for Research and Development Institute, Bucharest, Romania.

A constructed wetland is an artificial wetland created for the purpose of treating anthropogenic discharge such as municipal or industrial wastewater. It may also be created for land reclamation after mining, refineries, or other ecological disturbances. Constructed wetlands are engineered systems that use natural functions of vegetation to treat different water streams. Plants are often thought of as a treatment pathway. With a plant mono-culture, it is possible to accurately and to model the behavior of a wetland. In additional, a detailed water characterization is necessary to determine explanatory parameters and inventory the constituents in the water. In this paper, we provide our researches carried out in laboratory level, concerning the use of some selected plants in order to be used for the remediation of the wastewater resulted from the pharmaceutical industry. The tested species were the algae Chlorella vulgaris and the aquatic plants Lemna minor and Spyrogira sp. The algae culture was obtained by successive selection works starting from an identified cell and then all plant species were kept on special selected growth media. The 5 tested solutions used as representing some natural waste waters consisted in NH4NO3, Pb (NO3)2, MgSO4, ZnSO4 and NaCl. The ions which were analyzed concerning their concentration's evolution during a period of 72 hours (starting from 1%) were: Pb, NO3, SO4, Mg, Zn and Cl. The results indicate a better final situation in the case of those 2 aquatic plants, than in the case of using Chlorella.

Murtada A Oshi is pursuing his PhD at college of Pharmacy, Pusan National University, South Korea majoring in Manufacturing Pharmacy. His study in the field of colon-specific delivery of nano and microscale drug delivery system for the treatment of inflammatory bowel disease: ulcerative colitis and Crohn's disease. He is mainly focusing on solving out the disadvantages of traditional anti-inflammatory drugs, e.g. systemic side effects, poor targetability etc., used for the treatment of inflammatory bowel disease. Now, he is studying the anti-inflammatory activity of different nano and microscale drug delivery systems loaded with of anti-inflammatory drugs for the treatment of inflammatory bowel disease. He evaluates the anti-inflammatory activity of the formulations both in vitro and in in vivo. For in vivo study of inflammation, he used experimental animal colitis using different models such as dextran sodium sulfate, dinitrobenzene sulfonic acid and trinitrobenzene sulfonic acid.

Layer-by-layer (LBL) coating has gained popularity for drug delivery of therapeutic drugs. Herein, we described an approach for enhancing the therapeutic efficiency of the locally administered dexamethasone (Dx) for the treatment of inflammatory bowel disease (IBD). We utilized a LBL-coating technique for alternative coating of Dx microcrystals (DxMCs) with multiple layers of polyelectrolytes composed of poly (allylamine hydrochloride), poly (sodium 4-styrene sulfonate) and Eudragit® S100. The successful deposition of the layers onto DxMCs surfaces were confirmed through zeta potential measurement and confocal laser scanning microscopy, while the surface morphology was investigated through scanning electron microscopy. The drug encapsulation efficiency for LBL-DxMCs was 95% with a mean particle size of 2 µm and negative surface charge of -45 mV. Moreover, in vitro drug release studies showed a minimum release of the drug ( 15%) at an acidic condition during initial first 5 h followed by sustained-release at alkaline condition. For in vivo study, LBL-DxMCs were administered orally to male ICR mice suffering from dextran sulfate sodium-induced colitis. LBL-DxMCs was found to substantially enhance anti-inflammatory efficacy of the drug compared to uncoated DxMCs. Macroscopic, histological and biochemical (tumor necrosis factor-α, interleukin-6 and myeloperoxidase) examinations revealed marked improvements of colitis signs in the mice treated with LBL-DxMCs compared with those treated with uncoated DxMCs. Overall, the obtained results demonstrate that LBL-DxMCs are an effective and safe colon-targeted delivery system for the treatment of inflammatory bowel disease.

Angela Casarica has her expertise as a Project Responsible of different Romanian Research Projects related to Bacterial Cellulose for pharmaceutical and industrial purposes, studies to obtain an eyewash technology based truffle extract, fungal chitosan: isolation and biological characterization, curdlan-type polysaccharide obtained using a strain of Agrobacterium rhizogenes, product of natural origin for ophthalmic treatment and obtaining procedure. This work and experience in biotechnology was attained by approximately 10 biotechnological processes, seven patents and over 50 scientific papers published in specialized journals. She is a PhD in the field of Horticulture and she has obtained more medals at the International innovation fairs from Brussels and Geneva during the period 2008–2014. She is working as a Senior Scientific Researcher at the National Chemical-Pharmaceutical for Research and Development Institute, Bucharest, Romania.

Levans are fructose polymers synthesized by a broad range of microorganisms and a limited number of plant species as non-structural storage carbohydrates and they have potential applications in the pharmaceutical, food, and cosmetic industries. The present study shows a comparative analyses of polysaccharide type Levan biosynthesis in static and shaking fermentation by using Z. mobilis ATCC 10988 strain. All fermentation processes were carried out in Erlenmeyer flasks presenting a capacity of 500 ml and working volume of 100 ml culture medium, in both fermentation types: static and on the rotary shaker, with stirring of 220 rpm, with the temperature maintenance at 32°C for 72 hours. In figure 1, we can find the results obtained by static fermentation for different initial concentrations of sucrose. The best development of the microbial culture in terms of microbial biomass was seen for the initial concentration of 15% sucrose, due probably to the favorable ratio between the source of C and N. The largest amount of polysaccharide was obtained from the experiment with 40% initial sucrose (8.9 g%), but this value is also similar to the experiment with 25% initial sucrose (8.47 g%). The Figure No. 2 shows the results obtained by the stirred fermentation with the strain Z. mobilis ATCC 10988 for different initial concentrations of sucrose. Concerning the content of polysaccharide, if the initial concentration of sucrose in the biosynthesis medium was greater than 20%, the amount of polysaccharide was about 5 g%, without any increase of production in the case of higher concentrations. By comparing the results shown in figures 1 and 2, it can be noticed that the strain Z. mobilis ATCC 10988 performs better concerning the polysaccharide biosynthesis in the frame of a static fermentation, which is not a surprise considering that these bacteria are an optional aerobic one.

Corina Bubueanu is a Biochemist, has completed her PhD in Horticulture (2013) with 15 years’ of experience in research in the field of obtaining of new, natural medicines/drugs based on the selective herbal/vegetal, mushrooms extracts, involving both, fundamental research in order to design the most properly phytochemical composition, as well as applicative research concerning the following activities: the isolation of the various selective vegetal extracts enriched in the interest phytochemical compounds; analytical screening of the obtained vegetal extracts and the selection of the most proper ones by using specific, combined methods (spectral and spectrophotometric methods combined with HPTLC); the obtaining of the final pharmacological active product with pharmacological potential by the combination of the most active vegetal extracts; the setting up of the most appropriate and reproducible extractive technologies of these extracts and the correspondingly final active product as to the technological transfer from laboratory to pilot level.

The genus Lamium contains about 40 species, distributed in Europe, Asia, and Africa, some of which are used in traditional medicine. Currently, the genus Lamium is less studied and much less utilized compared to other members of the Lamiaceae family to which it belongs. Lamium album and Lamium purpureum are species belonging to the Lamium genus. Aerial parts of the two species and roots of Lamium album have applications in human and veterinary traditional medicine. Literature data presents as main components of the Lamium genus species iridoid glycosides. The active principles have diuretic, anti-inflammatory, anti-diarrheal, astringent, expectorant, vasoconstrictor, antirheumatic, haemostatic and emollient properties. In this paper, haemostatic properties of the Lamium species are investigated by two experimental (topical and systemic administration) models: hemostatic test by tail bleeding time detemination and acenocoumarol-carrageenan test. The haemostatic test results by tail bleeding determination topical administration, have demonstrated that both extracts have haemostatic activity. In the acenocoumarol-carrageenan test, systemic administration, only Lamium album extract have shown haemostatic activity, comparable with those obtained for administration of vitamin K. Based on the qualitative chemical composition in iridoid glycosides (HPTLC analysis) and the results obtained in experimental tests, there is the possibility that the compound responsible for the haemostatic activity is 8 acetylshanzhiside methyl ester. Both extracts have no toxicity based on an acute toxicity test.

Howard Lee is the Founder and Director of the Center for Convergence Approaches in Drug Development (CCADD). He serves as a Professor at the Department of Transdisciplinary Studies, Graduate School of Convergence Science and Technology, Seoul National University. He is also appointed at Seoul National University College of Medicine and Hospital, affiliated with the Department of Clinical Pharmacology and Therapeutics. He previously served as the Head of Global Strategy and Planning, Clinical Trials Center, SNUH. As of August 2017, he was appointed as Chair of the Graduate Program in Clinical Pharmacology, Seoul National University. He has spearhead the introduction of Accelerator Mass Spectrometry (AMS)-enabled exploratory early clinical drug development studies to the Korean biopharmaceutical R&D sector, which has awarded him two government grants.

In clinical drug development, it is important to understand the absorption, distribution, metabolism and excretion (ADME) properties of a drug in humans. The micro-tracer study based on the accelerator mass spectrometry (AMS) is an ultrasensitive technique to obtain human ADME profiles with a negligible radiation dose. KD101 is a novel compound under development to treat obesity. The aim of this study was to investigate the absorption, metabolism and excretion properties of KD101 in obese subjects. A randomized, open-label, single-dose, one-treatment, one-period, one-sequence study was conducted in six males with a BMI ≥27, who received KD101 at 400 mg with 3.52 µg of [14C]-KD101 (180 nCi) in the fed state. Plasma, urine and feces samples were collected up to 288 hours post-dose for mass balance and metabolite profiling. Plasma concentrations of KD101 were determined using a validated GC method. Total radioactivity in the samples was determined using AMS. Safety and tolerability was evaluated based on vital signs, adverse events, clinical laboratory tests, and electrocardiography. All of the subjects completed the study with no clinically significant safety issue. Mean total recovery rate (range) was 85.21% (75.36-99.01%), consisting of 77.96% (68.31-92.33 %) for urine and 7.26% (5.91-8.51%) for feces, which differed greatly from the pre-clinical data. Oral absorption of [14C]-KD101 was rapid with the peak plasma concentration reaching at 5.83h post dose, which was consistent with the previous report. In the urine radiochromatogram, five large peaks were identified including a peak represented by the parent compound. KD101 is excreted predominantly through the urine in humans. Many of the excreted materials in the urine were considered metabolites. This study demonstrated effectiveness of the micro-tracer study enabled by AMS in humans to investigate the ADME property of KD101, which hugely differed from that seen in the preclinical animals.

Pedro Almirall has his expertise giardiasis pharmacotherapy and public health. He has been working for years as a Clinical Epidemiologist and has experience in research, evaluation, and teaching, both in Cuban hospitals and education institutions. He has published his research in international scientific journals and currently, he is pursuing his PhD.

Introduction: At present, 5-nitroimidazole compounds are the pharmacological cornerstone for people with Giardia infections. However, treatment failures are increasingly reported. Mebendazole (MBZ), a benzimidazole compound, has received attention in treating patients with giardiasis since beneficial effects have been demonstrated in vitro and in vivo. Aim: The aim of this study was to assess in a systematic review and meta-analysis of randomized controlled trials (RCTs) the efficacy of MBZ compared to other antigiardial agents in children. Methods: RCTs of MBZ for the treatment of Giardia infections published in PubMed and EBSCO host were searched. Application of inclusion and exclusion criteria, data extraction, and assessment of methodological quality were independently performed in duplicate. The endpoint was the parasitological response to therapy. Results: Seven RCTs were found in the systematic review (639 patients) and were included in the meta-analysis. There is not clinical difference in the parasitological cure between MBZ and metronidazole (MTZ). The relative risk (RR) is 0.88 (95% CI 0.70-1.10), with a moderate heterogeneity (I2=66%). The prediction interval expected to cover the results of a new trial is wide enough (0.35-2.21) to support both a parasitological relevant difference favoring MBZ and a parasitological relevant difference favoring MTZ. Conclusions: This study synthesized available evidence on (and documented) the efficacy of MBZ in treating Giardia infection in children. There is no difference in efficacy between MBZ and MTZ regarding parasitological cure. Hence, the decision to support any of the competing treatments should be based nit in efficacy but, in other characteristics such as tolerance with the treatment or associated costs. Although our results suggest that MBZ may be an effective treatment option for children with Giardia infection, they should be interpreted and translated into clinical practice with caution, as the meta-analysis was based on a limited number of heterogeneous RCTs.

Hyun A Lee is a PhD student at the Department of Transdisciplinary Studies, Graduate School of Convergence Science and Technology, Seoul National University in South Korea. She is also a trainee in clinical pharmacology at the Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and Hospital. She has graduated from North Carolina State University (NCSU) in the USA with a Master’s degree in Biomathematics (2010). Prior to that, she went to the University of Alabama (2001-2004), where she received a Bachelor’s degree in Biology and Mathematics. After completing her undergraduate course, she worked as a Research Scientist at the Center for Chemical Toxicology Research and Pharmacokinetics (CCTRP) in NCSU. Her research area is physiologically based pharmacokinetic (PBPK) modeling and simulation to optimize new drug development. She has a lot of experience in PBPK modeling and simulation with new drugs.

YH4808 is a highly potent, selective and reversible potassium-competitive acid blocker on H+/K+-ATPase under development for the treatment of gastric acid related diseases. The objectives of this study were to develop a human PBPK model optimized by human pharmacokinetic (PK) data, to predict the PK profiles of YH4808 using the PBPK model in various settings and to mechanistically understand the main factor of clinically observed nonlinear PK of YH4808 by exploring various drug-drug interactions (DDIs). A PBPK model was developed using SimCYP® based on the physicochemical properties, preclinical in vitro and clinical data of YH4808 (Figure 1), which was further refined using human plasma concentrations obtained from a single-dose ascending phase I clinical trial of YH4808. The absorption of YH4808 was described by the advanced dissolution, absorption and metabolism model. The Vmax and Km values for each CYP isozyme involved in the metabolism of YH4808 were determined from the in vitro Ki values using a graphical method (GraphPad Prism7, GraphPad Software, Inc., USA). The DDI potential for YH4808 with other co-administered drugs was predicted using the PBPK model by calculating the geometric mean ratio of the model-predicted and observed for the area under the concentration-time curve (AUC). The PBPK model adequately predicted the observed concentrations of YH4808 after a single and repeated oral administration at 100 mg (Figure 1 (b), Figure 2 (a)). However, the simulated plasma concentration profiles after repeated oral administration at 200 and 400 mg were not in line with the observed concentrations, particularly toward the terminal phase, showing some sort of non-linear accumulation (Figure 2 (b) & (c)). The PBPK model-based simulated AUC of YH4808 increased by 2.08-2.90 times when co-administered with phenacetin, mephenytoin and dextromethorphan, respectively, suggesting the metabolism of YH4808 may involve CYP1A2, 2C19 and 2D6, which was confirmed by in vitro DDI studies. These metabolic pathways can be saturated at a higher dose of YH4808 at >200 mg, resulting in a non-linear PK profile. A PBPK model adequately predicted observed concentrations of YH4808 in humans after a single and repeated oral administration. The saturation of liver metabolism by CYP1A2, 2C19 and 2D6 appears to be associated with the nonlinear PK characteristics of YH4808 after multiple oral administration. The PBPK model can be used to predict the PK profiles of YH4808 in various clinical settings.

Roxana Madalina Stoica is a Researcher at the National Institute for Chemical-Pharmaceutical Research and Development-ICCF, Bucharest, Romania, in the Pharmaceutical Biotechnology Department. She has obtained her PhD in the field of Biotechnology and her research was focusing on the obtaining of biologically active principles from vegetal sources.

Bioactive compounds existing in the Capsicum genus, particularly capsaicinoids, provide many therapeutically uses due to its anti-inflammatory properties, to treat chronic pain, such as rheumatoid arthritis and neuralgia, and due to its anticancer, antimicrobial and antioxidant activity. The pungent metabolites in the fruits of Capsicum species are called capsaicinoids, and among the most abundant of these components are capsaicin and dihydrocapsaicin, which are responsible for about 90% of total pungency. Other important components resulting from the extraction of oleoresins are carotenoids (mainly capsanthin and capsorubin), which are widely used in the food industry, both because of their coloring and their antioxidant characteristics. The amount of capsaicinoids in peppers varies depending on the stage of maturity, variety used, cultivation conditions, nutrients soil and water stress. The capsaicinoids begin to accumulate in the early stages of fruit development and they reach a maximum rate as the fruit matures. The main objective of this study had involved the extraction of capsaicinoids from three Capsicum varieties, such as Guindilla Larga Roja, Fresno and Congo Trinidad, in order to obtain a pharmaceutical product for topical use. The realization of a topical formulation containing capsaicin, known for its analgesic action and local vasodilator, is a viable alternative in pain therapy. Capsaicinoids were identified in all extracts with concentration ranging from 0.3 to 0.7% (dry weight) and the best results were obtained with 96% ethanol as solvent comparing with methanol and acetonitrile, using Soxhlet extraction.

Eng. Georgiana IORDACHE has her expertise as a Project Responsible of different Romanian Research Projects related to Heparin Sodium for pharmaceutical and industrial purposes. This work and experience in biotechnology was attained by approximately 10 biotechnological processes, 2 patents and over 3 scientific papers published in specialized journals. She is a PhD student in the field of Biotechnology. She is working as a junior researcher at the National Chemical-Pharmaceutical for Research and Development Institute, Bucharest, Romania.

Levans are fructose polymers synthesized by a broad range of microorganisms and a limited number of plant species as non-structural storage carbohydrates and they have potential applications in the pharmaceutical, food, and cosmetic industries (1-5). This study presents a comparative analysis of the growth and consumption curves of Z. mobilis NCIB 1163 and Z. mobilis ATCC 10988, levan producing microorganisms. The growth and consumption curves were performed in a liquid medium with a concentration of 5% sucrose and 5% glucose. Thus, the bacterial cells in the exponential growth phase were centrifuged at 12.000 g and the pellet was washed twice with a sterile 0.9% NaCl solution. Finally, the cells were resuspended in 1 ml of physiological saline and they were used as inoculum in 5% liquid medium, which was monitored concerning the fermentable carbohydrate consumption and the growth. The consumption curves were performed under anaerobic conditions (the culture medium was coated with a layer of sterile paraffin oil) by periodical weighing of the medium seeded with different bacterial strains of Z. mobilis and by the graphical representation of weight loss (due to release of carbon dioxide). Exponentially growing cells were used as the inoculum, made at approximately 107 cells/ml. Cell growth was assayed turbidimetrically at a wavelength of 600 nm. The consumption curves of Z. mobilis NCIB 11163 and Z. mobilis ATCC 10988 bacterial strains on complete sucrose substrate medium, 5% under anaerobic conditions (Figure 1) revealed that strains NCIB 11163 and ATCC 10988 exhibit a similarly kinetics of consumption's substrate. The consumption curves of Z. mobilis NCIB 11163 and Z. mobilis ATCC 10988 bacterial strains on complete glucose substrate medium, 5% under anaerobic conditions (Figure 2) show that glycolysis is more intense than hydrolysis of sucrose. From the curves profile it is observed that the strain ATCC 10988 shows a curve having a more pronounced linear decrease.

Dr. Howard Lee is the Founder and Director of the Center for Convergence Approaches in Drug Development (CCADD). Dr. Lee serves as a Professor at the Department of Transdisciplinary Studies, Graduate School of Convergence Science and Technology, Seoul National University. Dr. Lee is also appointed at Seoul National University College of Medicine and Hospital, affiliated with the Department of Clinical Pharmacology and Therapeutics. Dr. Lee previously served as Head of Global Strategy and Planning, Clinical Trials Center, SNUH. As of August 2017, Dr. Lee was appointed Chair of the Graduate Program in Clinical Pharmacology, Seoul National University. Dr. Lee has spearhead the introduction of Accelerator Mass Spectrometry (AMS)-enabled exploratory early clinical drug development studies to the Korean biopharmaceutical R&D sector, which has awarded Dr. Lee 2 government grants.

Microtracing/microdosing is an innovative technology that can revolutionize the current paradigm of clinical drug development. Typically, a very small amount of the drug, i.e., ‘microdose’, which is less than 100 micrograms (or 30 nmoles for proteins), is administered to humans. Since this is much smaller than 1/100 of the pharmacologically active dose, microtracing/microdosing technology can be employed at a very early stage of clinical drug development even when there is limited animal toxicology data. Furthermore, in order to trace minute doses, an accelerator mass spectrometer (AMS) is required and the compound should be labeled, typically with 14C. The microtracing/microdosing study allows clinical drug development scientists for generating the intravenous pharmacokinetics, mass balance, metabolite profiling, and absolute bioavailability data much easier, faster, and at a significantly lower cost. Based on this understanding, this study investigated the current status and employment of AMS-based microtracing/microdosing studies in actual drug development. To achieve this objective, we performed an extensive search of the literature and public information, Delphi focus group interviews, surveys, and personal communications with the key players in the field. The number of the clinical studies that used 14C and AMS dramatically increased from only 3 in 2001-2005 to 59 in 2011-2015. The survey showed that 31.6% of new drug development scientists were planning to perform microtracing/microdosing studies. Furthermore, 73.7% of survey responders replied that they would consider AMS-based microtracing/microdosing studies if there is a well-established service provider. This study confirmed that the frequency of AMS-based microtracing/microdosing studies for drug development has been in a steady increase for the past decade or so. This increase was partly because several issues of AMS application in the previous era, such as dose-linearity, sample pre-processing, and high cost, have been adequately addressed. In conclusion, AMS-based microtracing/microdosing studies have been steadily employed in actual drug development, which is expected to increase further in the future.

Dr. Howard Lee is the Founder and Director of the Center for Convergence Approaches in Drug Development (CCADD). Dr. Lee serves as a Professor at the Department of Transdisciplinary Studies, Graduate School of Convergence Science and Technology, Seoul National University. Dr. Lee is also appointed at Seoul National University College of Medicine and Hospital, affiliated with the Department of Clinical Pharmacology and Therapeutics. Dr. Lee previously served as Head of Global Strategy and Planning, Clinical Trials Center, SNUH. As of August 2017, Dr. Lee was appointed Chair of the Graduate Program in Clinical Pharmacology, Seoul National University. Dr. Lee has spearhead the introduction of Accelerator Mass Spectrometry (AMS)-enabled exploratory early clinical drug development studies to the Korean biopharmaceutical R&D sector, which has awarded Dr. Lee 2 government grants.

The large majority of drugs and not volatile and amenable to standard analytical techniques. Volatile compounds pose a different set of challenges, requiring changes to the total sample preparation procedure and often analytical method. We encountered such situation during a human microtracer study (180 nCi human oral dose) of compound KD101, a single isomeric oil, (-)α-Cedrene (206.23 g/mol). A total of 6 subjects were administered 400 mg of unlabeled KD101 admixed with 180 nCi of radiolabeled drug as a tracer. Detection of the radiolabel was performed using graphite based Accelerator Mass Spectrometry (AMS), with support from Liquid Scintillation Counting (LSC) for early urinary time-points. Post- administration, cumulative urine and fecal voids were collected for 288 hr post-dose. Serial blood collections were taken frequently post-administration and then daily for the duration of the study. Preliminary work showed the KD101 could be completely removed under vacuum concentration in the absence of any trapping agent or biological matrix. This problem was exacerbated by the AMS processing method where samples are also evacuated under vacuum for torch-sealing inside quartz combustion (oxidation to gaseous carbon dioxide, prior to reduction to graphite). We found an acceptable technique to limit volatility losses through the pretreatment of all samples with an excess of tributyrin prior to concentration. The tributyrin served thus as both a carbon source and a chemical trapping agent. Sucrose was also tested but showed little ability to “capture” the compound during dry-down. We showed that parent compound recovery could be improved from <10 to 89% recovery using 2.0 mg of tributyrin per sample or LC fraction, and a minimum dry-down time (20 min). The overall results of the study validated the dilution method. Mean mass balance recovery was 85.21% (77.96% in urine, 7.26% feces). This is considered a good mass balance recovery given the fact that it was difficult to control for losses due to compound volatility. The concentration decay in the plasma was largely bi-exponential after the absorption period, with a wandering baseline out to 288 hr post-dose, with occasional “jumps” in concentration, which was attributed to the displacement of compound in fat depots. Metabolite radiochromatograms from urine and plasma showed intact parent in plasma with metabolites exclusively explaining the urinary components. In summary, we achieved satisfactory results for the mass balance and metabolism of a volatile oil using a chemical trapping agent. The detriment was a lowering of overall detection sensitivity (e.g. 0.88 dpm/mL of plasma vs. 0.05 dpm/mL of unmodified plasma), but sensitivity was still sufficient to achieve mass balance and perform metabolite profiling using highly sensitive AMS detection.