UDK 615 (497.11) ISSN 2217-8767 (Online) ARHIV ZA FARMACHUU Special Issue Book of Abstracts 11" Central European Symposium on Pharmaceutical Technology September 22 — 24, 2016 Belgrade, Serbia JOURNAL ION OF SERBIA 66 E September 2016 Seinen Gemeente ate mien =ARHIV ZA FARMACIJU CASOPIS SAVEZA FARMACEUTSKIH UDRUZENJA SRBIJE, ARCHIVES DE PHARMACIE - ARCHIVES OF PHARMACY IZLAZLOD 1951. GODINE IZDAVAC/ PUBLISHER, SAVEZ FARMACEUTSKIH UDRUZENIA SRBIJE / PHARMACEUTICAL ASSOCIATION OF SERBIA 11000 Beograd, Bulevar vojvode Mitiéa 25, pot. fah 664 tel/fax: + 381 11 2648 385; ~381 11 2648 386 e-mail: fds@sbb ss; sfus@farmacijaorg ‘wwe farmacijaorg IZDAVACKI SAVET / PUBLISHING COUNCIL Milana Dutié -Apotsia ,Beogr Sonja KuStrin-Dordevie- Uduzenje maceu Beograd, Ivanka Miletié - sever famaceuskih niena Sie, Diubravka UroSey - Saver annacelskinudndenj Sebi, [Nenad Vulovié - Uanzenje farmaceua Beopada UREDNICA ARHIVA/ EDITOR-IN-CHIEF Marija Primorae Univeraitetw Beogeado -Farmacetki okt, Katedra a fammaceushu tehaoog i kezmetlogi ZAMENIK GLAVNOG UREDNIKA / DEPUTY EDITOR Radica Stepanovié-Petrovie Universit w Beograd Famaceuis fit, Katie a frmakoonia LEKTOR / LECTOR Mirjana Bla?ié-Mikié Radove objavijene u Zasopisu Arhiv za farmaciju indeksiraju: EMBASE i SCOPUS. ARHIV ZA FARMACUIU izle fest puta godiinje a sajtu Saveza farmaccutskih udruzenje Srbije www farmacija.orgUREDNISTWO/EDITORIAL BOARD Antonijevié Biljana, Univerzteu Beogradu~ Farmaccusk fakultet, Katedra za toksikologiu 2a tksikologiu - akademik Danilo Soldatovié, Couladis Maria, Department of Pharmacognosy & Chemisty of Natural Products, School of Pharmacy University of Athens, Gece, Durovié Dragan, ‘Agencia za lekove i medicinsa sredsva Sibi, Beogr Harding Ljiljana, Global Manufacturing Serves at Pier Pharmaceutical, Peapack, New Jersey, USA, Jovanovié Aleksandar, Divison of Medical Senses, Medical School, University of Dundee, UK, Kovatevié Nada, Universitet u Beograd ~Farmaccutsh fakultet, Katedra za farmakognoni, Kowalska Teresa, Institut of Chemisty, Silesian University, Katowice, Poland, Leposavié Gordana, Univesteu Beograda~ Farmaccusk fakultet, Katedra 78 fiotgi, ‘Milenkovié Marina, ‘Univesteu Beogradu- Famaccutsk fake, Kate Petrusevska-Tozi Lidij Institute for Applied Biochemistry, Faculty of Pharmacy, University Ss Cyril and Methodus, Skopje, R Macedonia, Primorac Marija, Univer Beograd - Frmaceutsi kt, Katedra a farmaceutsk tehnolgia i kozmetologiy, Spasojevié-Kalimanoyska Vesna, ‘Universitet u Beograd ~ Farmaceus fakultet, Katedra za modicinsku biohemiju, Srdié Stanko, Pharmaceutical Technology Department, The Faculy of Pharmacy, Univers of Ljubljana, Slovenia, Stankovié Ivan, Universite u Boogradu~ Farmaccusifakultel, Kaede 2a bromatologii Stepanovié-Petrovié Radica, Univrztetu Beogred- Farmaceulsk fakultet, Katedra 2a farmakooei, Stojanovié Biljana, Universitet u Beograd ~Farmaceus fakultet, Katedra 2. ‘Tamburié Slobodanka, Schoo! of ManagementAScience, London College Tasié Ljiljana, Universitet u Beograd - Farmaccutsk fakultet, Katdra za scion farmaci i farmaccusko zakonodavsvo Viadimiroy Sote, Univerztetu Beogredu~Farmacctsk fakultet, Katedra 2 farmaceusku hemi, Vutiéevié Katarina, Universitet u Beogredy~Farmaceus fake, Katedra za frmakokineti i Minigka tarmac, Vuleta Gordana, ‘Universitet u Beograd -Farmaccusk fakultet, Katedra za farmaccusku tehnologiu i kormetologit a mikrobiologiu i imunologiy, tk ekova, ashion, University ofthe Arts, London, UK,Guest Editors’ Preface 11° Central European Symposium on Pharmaceutical Technology is focused on four mean themes: (I) From smart materials to advanced drug delivery systems, (2) Regulatory science - from generics to biosimilars, (3) Pharmaceutical engineering and (4) In vivoyin vitrofin silico modelling. The authors of 2 plenary lectures, 9 invited lectures, 25 oral and 121 poster presentations have met and exceeded the organizers’ expectations with the quality and variety of their contributions. Recent advances in drug delivery are presented, using very new or improved existing smart” materials. These materials were characterized from pysicochemical characterization to in vitrofin vivo evaluation, Furthermore, advances in drug delivery were presented such as 3D printing devices, and macro- and nanosized drug delivery systems. Special attention has been paid to pharmaceutical engineering with emphasis to continuous manufacturing and Process Analytical Technology tools implementation in pharmaceutical manufacturing, Regulatory aspects of drug and formulation development are shown, both for biologics and generics. Moreover, quality by design aspects in development of drug delivery systems are evaluated from regulatory and academic point of view. In In vivo/in vitro/in silico modelling session, various aspects of quantitative- structure permeability relationship modeling, bioperformance prediction of oral drug products, computer simulations of iontophoresis in the drug delivery system and particle deposition in nasal cavity are presented. Over 280 delegates from 30 countries give this symposium a truly intemational character and represent a strong basis for creative scientific interactions and wide dissemination of new concepts. The organizers are grateful for all the assistance from the supporting societies and associations in promoting the idea of CESPT in the international community. Prof’S. Ibrié, President of CESPT Prof A. Mrhar Co-President of CESPTOP04 EFFICACY ASSESSMENT OF SELF-ASSAMBLED PLGA-PEG-PLGA NANOPARTICLES: CORRELATION OF NANO-BIO INTERFACE INTERACTIONS, BIODISTRIBUTION, INTERNALIZATION AND GENE EXPRESSION STUDIES Ss. Dimehevska's, N. Geskovski', > R. Koligi', N. Matevska-Geskovska’, V. Gomez Vallejo’, B. Szezupak’, M. Errasti Lopez’, E. San Sebastian’, J. Lop’, D. R. Hristov’, M. P. Monopoli’, Dimovski’, K. Goracinova "Institute of Pharmaceutical technology, Faculty of Pharmacy, UKIM, Skopje, R. Macedonia Center for Pharmaceutical biomolecular analyses, Faculty of Pharmacy, UKIM, Skopje, R. Macedonia * Radiochemistry Platform, Molecular Imaging Unit, CIC biomaGUNE, San Sebastian, Spain * Molecular Imaging Unit, CIC biomaGUNE, ‘San Sebastian, Spain * Centre for BioNano Interactions, School of Chemistry and Chemical Biology, UCD, Dublin, Ireland INTRODUCTION The aim of our study was to attempt to introduce the concept of quality by design and safety by design in the development of biodegradable self-assembly nanoparticles (NPs) as carriers of extremely hydrophobic Arh, farm. 20 6 Speil e anticancer drug SN-38. Beside the development and optimization of several formulation properties, we employed battery ‘of assays in order to study NP interactions with the biological environment in an in vitro and in vivo setup. MATERIALS AND METHODS. Materials PLGA-PEG-PLGA —_triblock copolymers (DLLA:GA=I:1, Mw~70:8:70kDa and DLLA:G. Mw~6:10:6kDa)_—_ were purchased from Akina Inc, USA. 7- Ethyl-10- hydroxy camptothecin (SN-38) was obtained from Xi*An Guanyu Bio-tech Co., China, Lutrol@F127 was kindly donated by BASF, Germany. Bovine serum albumin (BSA) was acquired from Sigma Aldrich, USA. am was eluted from ”"Mo/"Te generator (Cis Bio Intemational, France). All other chemicals were of analytical grade and were used as received. ‘Methods Experimental design preparation D-optimal designs, nanoprecipitation and two types of copolymers PLGA-PEG-PLGA (DLLA:GA=I:1, — Mw~70:8:70kDa_— and DLLA:GA~7:3, Mw~6:10:6kDa) were used for optimization of _polymer/drug/ Lutrol®@F 127 concentration ratio in order to maximize the drug loading and provide appropriate particle size for passive targeting. and nanoparticle Physico-chemical characterization of NPs Particle size, size distribution and zeta potential of the NPs were measured using DLS. HPLC method was used for quantification of encapsulated SN-38. Morphological analysis of NPs was performed using Teenai G2 20 S-Twin 200 kV TEM. Protein corona analysis Bradford protein assay was used for quantitative analysis of BSA adsorption Spectroscopic characterization of NP-BSA interactions was carried out using FTIR. Semi-quantitative analysis of adsorbed proteins was performed using SDS PAGE atfarm. 2016;66 / Special Issue several time points after NP incubation with 50% human plasma. SW480 cell culture studies Internalization of fluorescently labeled NPs ‘was quantified using flow cytometry. In vitro cytotoxicity studies were performed with blank NPs dispersed in cell culture medium using MTT assay. Gene expression assay was conducted by PCR on UBD, RGCC, FGF3 and HIST genes, Biodistribution studies Direct “Te radiolabeling of NPs was performed using sodium dithionite as a reducing agent(1). Biodistribution and blood circulation time was evaluated in healthy Wistar rats using SPECT imaging and gamma counting. RESULTS AND DISCUSSION It was confirmed that the core composition, SN-38 concentration and the amount of surfactant affects the drug loading and particle size. Two optimal formulations were selected (NPI — high Mw polymer, NP2-low Mw polymer) for further studies. Maximum drug content in the optimized formulations was around 6% while the particle size was in the range of 70-100 nm, Zeta potential measurements have shown that NPI is slightly more negatively charged compared to NP2, most probably because of its higher PLGAJPEO ratio and shorter PEO loops. TEM images pointed to spherical shape of the NP formulations. Quantitative BSA adsorption assay demonstrated lower affinity of NP2 towards BSA compared to NPI, probably due to the differences in the PEO density and chain length at the NP surface. The Amide I derivation has shown that the secondary structure of BSA was preserved for more hydrophobic nanoparticles (NPI) pointing to the presence of strong hydrophobic interactions between BSA and NP core. Plasma protein adsorption studies have shown that initially, most abundant plasma proteins were adsorbed on both types of NPs and in the later stage they were replaced by proteins with higher bindi NP surfaces Cytotoxicity studies revealed the significant effect of the presence of serum proteins in the culture medium upon the growth rate of SW- 480 cells. Also, the internalization was significantly decreased for NP2 in the presence of proteins (FBS). Increased expression of UBD and RGCC genes and decreased expression of FGF3 and HIST genes for both formulations was observed, which corresponds to the pattern of pharmacological activity of SN-38. Plasma profile curves and_biodistribution studies pointed to evident differences in the circulation time among the — evaluated formulations (Figure 1). affinity towards Al BI Fig. 1 SPECTICT fulbhody sane of he wy ibNrD CONCLUSIONS, Differences in physico-chemical properties induced formation of protein corona with different composition which further affected biological behavior; cytotoxicity, internalization and biodistribution of the prepared polymeric NPs. ACKNOWLEDGEMENT ‘This research received support from the FP7 QualityNano Project. REFERENCES |.Geskovai N, KuzmanovlaS, Simonosta Creaeva M, Cals, Dimchesha', Pessoa M, ea. Comparative bedsebuson suis of technetium radolabsled arphighlie nanoparticles sng tree diferear reducing agents during he bcing Procsdare J Labelled Comp. Rasiphrm, 20.5518) 589.95