16^th International Conference and Exhibition on Pharmaceutics & Novel Drug Delivery Systems March 19-21, 2018 Berlin, Germany

Zhihui Hao has completed her PhD from China Agricultural University and Post-doctoral studies from Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS. She has published more than 30 papers in reputed journals and has been serving as an Editorial Board Member in journals of repute.

The short elimination half-life of cefquinome limits its use for treating lung infections caused by Klebsiella pneumonia in veterinary clinic. The aim of this study was to prepare cefquinome-loaded poly lactic-co-glycolic acid (PLGA) microspheres and to evaluate their in vitro and in vivo characteristics and pharmacodynamics for therapy of pneumonia in a rat model. Microspheres were prepared using a spray-drying method and were characterized in terms of morphology, size, drug-loading coefficient, encapsulation ratio and in vitro release. The prepared microspheres were spherical with smooth surfaces and uniform size (12.4±1.2 μm). The encapsulation efficiency and drug loading of cefquinome was 91.6±2.6% and 18.3±1.3%, respectively. In vitro release of cefquinome from the microspheres was sustained for 48 h and was supported by the Korsmeyer–Peppas model. In vivo studies identified the lung as the target tissue and the region of maximum cefquinome release. A partial lung inflammation was observed, but disappeared spontaneously as the microspheres were removed through in vivo decay. We have successfully prepared a drug-loaded microsphere delivery system for the treatment of pneumonia caused by Klebsiella pneumonia ATCC 10031. The sustained cefquinome release from the microspheres revealed its applicability as a drug delivery system that minimized exposure to healthy tissues while increasing the accumulation of therapeutic drug at the target site. Our study highlights the targeted drug delivery of cefquinome as the promising alternatives to control the important zoonotic pathogens.

Zhihui Hao has completed her PhD from China Agricultural University and Post-doctoral studies from Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS. She has published more than 30 papers in reputed journals and has been serving as an Editorial Board Member in journals of repute.

Pidotimod (PDM) represents a new class of biological response modifiers. It enhances immunological activity of both innate and adaptive immune systems. In vivo studies in animals and humans have demonstrated that PDM has immunoprotective activities. The aim of this study was to prepare PDM soluble powder and to investigate the immune enhancement properties of PDM in chickens vaccinated with Newcastle disease virus (NDV). The chickens were averagely divided into six groups, except black control group, which was vaccinated with NDV. At the same time of the vaccination, the chickens in three PDM groups were given water with PDM for five days, with the PDM at low, medium and high concentrations (0.25 g/L, 0.5 g/L, 1 g/L), respectively. The control drug group was treated with 0.2 ml/PDM dose via drinking water, in vaccination and black control groups, with equal volume saline, once a day for five successive days. On days 14, 21 and 28 after the vaccination, the growth performance, the lymphocyte proliferation, serum antibody titer, the CD4/CD8 cell ratios, interleukin-2 (IL-2) and interferon-gamma (IFN-γ) were measured. PDM at suitable doses (0.5 g/L) could significantly promote growth performance, lymphocyte proliferation, enhance serum antibody titer, CD4/CD8 cell ratios and improve serum IL-2 and IFN-γ concentrations. It indicated that PDM could significantly improve the immune efficacy of Newcastle disease vaccine using doses of 0.5 g/L, these results are consistent with the drug acting as an immunopotentiator.

Julia Chudzian is a PhD student in Molecular Biology at the Faculty of Biotechnology, Department of Protein Engineering at the University of Wrocław, Poland. She has her expertise in phage display technology, as well as in engineering proteins and mammalian cell culture. Her major interests are in targeted anti-cancer therapy, particularly antibody fragments and antibody engineering.

Fibroblast growth factor 1 (FGF1) and its receptors (FGFRs) regulate crucial biological processes like cell proliferation and differentiation. Aberrant activation of FGFRs by their ligands can promote tumor growth and angiogenesis in many tumor types including lung or breast cancer. The development of FGF-targeting molecules with potential implications for the therapy of FGF-driven tumors is recently being considered a promising approach in the treatment of cancer. In this study, to selectively target FGF1, we have used phage display method and selected a panel of monoclonal antibody fragments in scFv format (single-chain fragment variable) originating from Tomlinson I and J libraries. We performed ELISA testing, scFv clones DNA sequencing and preliminary verification of an interaction with the antigen using Bio-layer interferometry (BLI) method. From 368 clones we chose seven displaying the most favorable features that were expressed in E. coli and purified. Biochemical and biophysical analyses have enabled characterization of the selected antibody fragments among which we chose one clone, named scFv0712, which demonstrated the highest affinity and specificity to FGF1. In the next step, we reformatted the best clone to the scFv-Fc format. scFv0712-Fc was produced in mammalian CHO cells, purified and characterized. With the use of BLI analysis we determined the binding parameters of both antibodies to FGF1. The value of the dissociation constant to the antigen of scFv-Fc exhibited a significant reduction in reference to the parental clone, what showed that the reformatting process has led to an improvement in the binding parameters of the antibody. Moreover, both scFv0712 and scFv0712-Fc non-negligibly reduced FGF1-stimulated activation of FGF signaling, as demonstrated by their suppression of Erk1/2 phosphorylation. Generated antibody fragments serve as novel FGF1 inhibitors and can be further utilized as powerful tools to use in the studies on the selective cancer therapy.

Karolina Świderska is a Research Associate at the Department of Protein Engineering, University of Wrołcaw. She is also pursuing her PhD in developing new alternatives for antibody drug conjugates (ACDs) based on fibroblast growth factors (FGFs). Her research interest includes various bioconjugation techniques, selective targeting of drugs and resistance of cancer cells to conventional cytotoxic drugs.

Antibody drug conjugates (ADCs) are the most intensively developed class of anticancer therapeutics. ADC is composed of antibody that recognizes specific target presented on cancer cell and highly potent cytotoxic payload that kills effectively upon internalization into cancer cell. In this study, we have tested fibroblast growth factor 2 (FGF2), which is a natural ligand for FGF Receptors (FGFRs), frequently overexpressed in various type of cancers, to verify whether non-antibody molecule can be used as a targeting agent. Instead of commonly used cytotoxic agents like monomethyl auristatin E (MMAE), emtansine (DM1) and calicheamicin, we applied α-amanitin, a mushroom toxin which specifically inhibits DNA transcription that has been shown as an alternative payload in ADCs1. We produced FGF2-α-amanitin homogeneous conjugate via maleimide chemistry with determined amanitin:FGF2 ratio as 1:1. The FGF2-α-amanitin conjugate has shown improved antitumor activity against osteosarcoma and various types of lung cancer cell lines. The cell-killing effect was dependent on the presence of FGFRs on cell surface as FGFRs-negative cells did not show any increased morality. Here, we show that FGF2 can be used as an effective targeting molecule and α-amanitin can be applied as an alternative payload in ADCs.

Magdalena Lipok graduated from Opole University, Poland with an MSc degree in Chemistry. Currently, she is the student of PhD Studies in Molecular Biology at University of Wroclaw. The main topic of her PhD thesis focuses on finding FGF1-FGFR signalling pathway inhibitor, which potentially can be used in anticancer therapy. Her scientific interests are small molecule inhibitors, anticancer application of peptides and protein drug discovery and development in anticancer therapy.

Fibroblast growth factor 1 (FGF1) is a protein with a very high mitogenic activity. This polypeptide possesses ability to bind with high affinity to all isoforms of fibroblast growth factor receptor (FGFR), which are often overexpressed in different types of human cancers. Dissociation of FGF1-FGFR complex with specific low molecular weight inhibitors can be considered a new approach to anticancer therapy. Therefore by targeting at FGF1 we can prevent unwanted FGF1-FGFR interaction, which will decrease overall FGFR activation. To find FGF1 inhibitors we have used cyclic C7C Phage Display Peptide Library. We performed 3 rounds of in vitro biopanning using 96-well plates coated with FGF1. The enriched phages were analysed with ELISA assay. Fourteen clones that showed highest binding to FGF1 were sequenced and five selected peptides were synthesized, oxidized to form and purified on RP-HPLC. The proper molar mass of peptides was confirmed by MALDI-TOF mass spectrometry. We verified the ability to block FGF1-FGFR1 interaction by four selected peptides. The inhibitory effect of peptides was confirmed by blocking of in vitro FGF1-mediated activation of ERK1/2 pathway. Based on obtained data, we chose two peptides which possess the highest ability to inhibit FGF1-FGFR1 interaction. Currently we are working on reformatting selected peptides into peptibody format, to obtain more stable molecules with higher binding avidity to FGF1.

Ioana C Carlan has completed both her BS (Biochemical Engineering) and MS (Pharmaceutical and Cosmetic Products) at Gheorghe Asachi Technical University of Iași, Romania. She is currently continuing her studies with a Doctoral Program in the field of Chemical and Biological Engineering at the Faculty of Engineering from University of Porto, Portugal. Her research is focused on the microencapsulation of watersoluble vitamins using spray-drying technique. She has published two papers and has attended three conferences

Vitamin B1 represents an essential nutrient for human health, responsible for energy and metabolism. Therefore, it should be provided daily through a balanced diet. However, it is possible to develop a deficiency or diseases related to poor absorption. This usually affects persons who consume too much alcohol, pregnant women and some least developed countries. Pharmaceuticals and fortified food products with vitamin B1 microcapsules content may be a promising alternative for the prevention or treatment of deficiency. In this work, vitamin B1 was microencapsulated using spray-drying, a simple, flexible and economical technique that can overcome vitamin B1 stability problems. Several biopolymers were tested as coating materials, namely Arabic gum, carrageenan, chitosan, maltodextrin, modified chitosan, modified starch, pectin, sodium alginate and xanthan. Microcapsules with a 0.25% (w/w) of vitamin were obtained using a mini spray-dryer BÜCHI B-290 and the process showed acceptable values for the product yield: 20-50%. For the characterization of microcapsules in terms of size, a particle size analyzer was used and the morphology was determined by scanning electron microscopy. The size, shape and surface of each type of vitamin B1 microcapsules differ according to the type of biopolymer used as coating. Controlled release studies of vitamin B1 from the microcapsules were determined by spectrophotometric analysis with fresh and four months old samples, in two dissolution mediums: deionized water (20ºC) and simulated gastric fluid (37ºC). The obtained release profiles highlighted different behaviours: fast release, just a few minutes needed and slow release, up to several hours. Kinetic models, like zero order, first order, Higuchi, Korsmeyer-Peppas and Weibull were also applied to see which fits better. This research reported significant results and proved the success of microencapsulating vitamin B1 with different encapsulating agents. Also, it offered important information about the release profiles and stability over time.

Jung Dong Kim got a B.S. and Ph.D. degree in biotechnology from Yonsei University(South Korea). Dr. Kim has joined to Raphas at 2010 for the industrialization of microneedle using Droplet-born Air Blowing technology. And he was a cooperative researcher of Institute of Industrial Science, The University of Tokyo, since 2014 to 2017. He was also a steering committee of International Conference on Microneedles at 2016. He received IR52 Jang Young Sil Award twice from the Minister of Science, ICT and Future at 2016 and Minister of Science and ICT at 2017. His research is focused on the commercialization of dissolvable microneedle technology in cosmetic and medical area.

The microneedle-based transdermal delivery system has been developed to facilitate a minimal invasive self-administration method for cosmetic, medical device and pharmaceutical applications. Most dissolvable microneedles are fabricated by micro-molding method for the last 15 years. Therefore, Raphas developed a droplet-born air blowing (DAB) method that has some advantages in fast and stable fabrication of dissolving microneedles by drying polymer at room temperature in a few minutes. Additionally, DAB method provides precise control of drug dose in the microneedle by controlling dispensing amount. DAB-based dissolving microneedle products are already commercialized in various countries. This study showed the characteristics of 2 different structured dissolvable microneedles, which manufactured in our mass production system. First structure is single-layered microneedle. Second structure is double-layered microneedle. Various ingredients were loaded within these microneedles. The loaded amount of ingredient was analyzed by enzyme-linked immunosorbent assay (ELISA) or HPLC/UV system. Skin permeability of microneedle was confirmed by OCT (optical coherence tomography) and delivered amount of drug into the skin was analyzed using Franz diffusion cell (Logan, FDC-6T). In vivo OCT images clearly showed that whole length of microneedles could penetrate into human skin. In vitro and ex vivo studies using Franz diffusion cell showed excellent delivery efficacy compared to topical solution. DAB technology suggests a way to solve the problems of conventional micro-molding method to fabricate dissolvable microneedle. Based on the method, we have successfully developed mass production system to manufacture microneedle-arrayed patch. We will start clinical trials using this platform technology in this year.

Farah El Mohtadi is a Post-doctoral Research Fellow at The University of Manchester. She obtained her PhD in Pharmacy and Pharmaceutical Sciences from the University of Manchester in 2017. She earned her Master’s Degree in Pharmaceutical Technology from Jordan University of Science and Technology, and she joined the University of Manchester in 2013. Her research currently focuses on the development and characterisation of novel stimuli responsive polymers for drug delivery.

Enzyme replacement therapies (ERT) are used to treat lysosomal storage diseases (LSDs) and are some of the most expensive treatments available today. Examples of LSDs include Gaucher’s disease (GD), Hunter’s disease and Fabry’s disease (FD). GD is the most common of the LSD caused by the deficiency of glucocerebrosidase; a lysosomal enzyme that catalyzes the hydrolysis of the glycolipid glucocerebroside to ceramide and glucose. Current therapy relies on a bi-weekly injection of an exogenous glucosidase which costs approximately US$150,000–300,000 per year, making it one of the most expensive treatments available in the market. Efforts to improve the stability of the enzyme may therefore result in considerable benefits, both clinically for the patients and financially for the insurance companies/care providers. It has been found that oxidation can be a significant factor in reducing the activity of beta-glucosidase in those with GD. Moreover, in FD it has been found that anti-body formation in response to the exogenous enzyme can play a significant role in reducing the available enzyme as well as causing significant immunogenic side-effects. In the mind-set of improving the lifespan of these enzymes, we have developed an enzyme-polymer conjugate, specifically lysozyme (a model enzyme) and an anti-oxidant polysulfide. The polymer was synthesized via anionic ring opening polymerisation, and the conjugation was performed via the NHS-ester reaction to free amines (lysines) on the surface of lysozyme. The cytotoxicity of the synthesized polymer was compared to that of poly (ethylene glycol) (PEG) via MTS assay. The activities of the conjugates were performed on the lysozyme conjugates measuring the lytic activity towards fluorescein-labelled Micrococcus lysodeikticus. SDS-PAGE gel electrophoresis together with MALDI-TOF mass spectrometry analysis revealed formation of conjugates of 1–3 polysulfide chains per lysozyme with only a slight reduction in lysozyme activity. The synthesized polymer showed a toxicity profile virtually identical to PEG. Unlike the native lysozyme, the lysozyme-polysulfide conjugate was found to be resistant to oxidative denaturation and proteolysis.

N Basaran Mutlu Agardan graduated from Gazi University Faculty of Pharmacy, and obtained there her PhD from the Department of Pharmaceutical Technology. She then gained a scholarship from the Scientific and Technological Research Council of Turkey and completed her Post-doctoral research studies at the Center for Pharmaceutical Biotechnology and Nanomedicine, Northeastern University, Boston. Her area of research focuses on the smart drug delivery systems, liposomes, drug/gene delivery in cancer and oral absorption enhancement

Systemic invasive fungal infections are the leading causes of mortality and morbidity among immunocompromised patients such as, cancer patients undergoing chemotherapy, AIDS patients and recipients of solid organ transplants. Amphotericin B (AmB), a polyene group antibiotic is a common choice for life-threatening systemic fungal infections and parasitic diseases due to its broad spectrum of activity. AmB is a BCS class IV drug with low solubility and low permeability properties. Although amphotericin B is a very potent agent, clinical use is limited due to its narrow therapeutic index and side effects. Cyclodextrins (CDs) are a group of pharmaceutical excipients containing a lipophilic central cavity and a hydrophilic outer surface. In aqueous solutions, they have the ability to form inclusion complexes with many drugs by taking up a drug molecule/some lipophilic moiety of the drug molecule into their central cavity. The inclusion complexes are very successful at increasing the solubility of low-soluble drugs. The main goal of a drug delivery system is to enhance the therapeutic index and reduce side effects of encapsulated drug. Liposomes are the macro/nano sized vesicular drug delivery systems with well-established advantages. The aim of this study was firstly, preparation and characterization of inclusion complexes of AmB with CDs. Secondly, formulation development of AmB double loaded liposomes by loading AmB-CD inclusion complex to the inner hydrophilic compartment, and hyrdrophobic AmB to the outer phospholipid membrane. For this purpose, solubility studies were carried out by using three CD derivatives (α-CD, HP-β-CD and sulfobutyl ether β-CD). α-CD was found to be the most enhancing derivative of AmB solubility. AmB- α-CD complexes were prepared and characterized by DSC, FTIR, X-ray diffraction measurements. Design of experiment (DoE) is planned to assist the formulation of AmB and AmB- α-CD double loaded liposomes in order to obtain more comprehensive and comparable results.

The diffusion and adsorption behavior of cisplatin in silica nanopores is investigated using molecular dynamics simulation. Two different silica conformations are studied in order to characterize the influence of surface polarity. We find a strong influence of the pore diameter on the diffusion coefficient; only for pore diameters larger than roughly 1.6 nm, cisplatin assumes the same diffusion coefficient as in bulk water. Our results also allow estimating escape times of cisplatin from a pore. Acceleration techniques are used in order to sample adsorption phase space efficiently and to identify realistic adsorption conformations. We find major differences between the polar and nonpolar surfaces. Electrostatic interactions govern the adsorption on polar surfaces and can be described by the alignment of the molecule dipole with the surface dipole; hence, spreading of the molecule on the surface is irrelevant. On nonpolar surfaces, on the other hand, van-der-Waals interaction dominates inducing surface spreading of the molecule.

Marjan A Nejad is a PhD candidate in Kaiserslautern University in Germany. She is doing her research in Molecular Dynamic Studies in Drug Delivery System.

Saleh Mohammed Saeed Alamri completed his Master’s degree of Pharmaceutical Biotechnology from De Montfort University, UK in 2014. He is working as an Assistant Director Pharmacy of Material Management in Prince Sultan Military Medical City, KSA. He presented his work in national and international scientific conferences and meetings.

The structure of pharmaceutical granules is highly complex, with a morphology combining discrete solid, but physically variable particles, separated by air pores of various dimensions. The compression procedure for both granules and powders within the pharmaceutical industry is a process represented by four or five steps. Modern advances in various qualitative and quantitative techniques examining the internal morphological changes within powders and granules under subject to various compression loadings have been reported. However, such techniques tend to be costly, demanding significant expertise and highly specialised equipment. As an alternative, this research project has investigated the potential for simple colouring of pharmaceutical granules to monitor free-flowing, solid-state pharmaceutical substrates (i.e., powders or granules with dimensions of 420-800 µm within confined die geometry. This technique permits visualisation of granule behaviour and determination of bed density as granules are permitted to fill the die in multiple strata. Results demonstrate that the die wall has only a minor influence at the commencement of the compression cycle, where granules layers become homogeneously compressed. Gradual increased loading subsequently demonstrates a more heterogeneous distribution of coloured layers, where the base layer is shown to exhibit the least density changes. The liminal layer areas exhibit significant coloured granule diffusion.