2nd SPLC-CRS Young Scientists Meeting...2nd SPLC-CRS Young Scientists Meeting Novoa Santos...

Click here to load reader

  • date post

    04-Jul-2020
  • Category

    Documents

  • view

    1
  • download

    0

Embed Size (px)

Transcript of 2nd SPLC-CRS Young Scientists Meeting...2nd SPLC-CRS Young Scientists Meeting Novoa Santos...

  • 2nd SPLC-CRS Young Scientists Meeting

    Novoa Santos Auditorium, School of Medicine (USC)

    Rua San Francisco s/n

    Santiago de Compostela

    January 23rd, 2019

    BOOK of ABSTRACTS

    Rectorship of University of Santiago de Compostela Colexio de San Xerome, Praza do Obradoiro

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    1

    Contents

    Contents ....................................................................................................................... 1

    Organizing Committees ................................................................................................ 2 Welcome Message and Venue ..................................................................................... 4 Scientific Program ......................................................................................................... 6 Invited Speakers ......................................................................................................... 10 Abstracts: Oral presentations ...................................................................................... 13 Abstracts: Poster presentations .................................................................................. 29 List of authors ............................................................................................................. 63 List of co-authors ........................................................................................................ 67 List of participants ....................................................................................................... 72

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    2

    Organizing Committees

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    3

    SPLC-CRS Board

    Prof. Dr. Dolores Torres, President, University of Santiago de Compostela, Spain

    Prof. Dr. María Blanco Prieto, Secretary, University of Navarra, Spain

    Prof. Dr. Carmen Maria Évora García, Vice-President, University of La Laguna, Spain

    Prof. Dr. Bruno Sarmento, Director, University of Porto, Portugal

    Prof. Dr. Helena Florindo, Director, University of Lisbon, Portugal

    Prof. Dr. Manuel Santander, Treasurer, University of Castilla La Mancha, Spain

    Young Scientist Committee

    Alba Calvo Bacaicoa, University of Navarra, Spain

    Carlos Rodríguez, University of Navarra, Spain

    Flávia Sousa, University of Porto, Portugal

    Liane Moura, University of Lisboa, Portugal

    María Plaza, University of Castilla La Mancha, Spain

    Matilde Durán Lobato, University of Santiago de Compostela, Spain

    Patricia Costa Filipe, University of Lisbon, Portugal

    Sandra Jesus, University of Coimbra, Portugal

    Sonia Vicente-Ruiz Salvador, Centro de Investigación Príncipe Felipe Valencia, Spain

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    4

    Welcome Message and Venue

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    5

    Dear participants,

    On behalf of the Board of the Spanish-Portuguese Local Chapter of the Controlled Release Society (SPLC-CRS), we are delighted to welcome you to the 2nd SPLC-CRS Young Scientists Meeting at the Novoa Santos Auditorium, School of Medicine, University of Santiago de Compostela, on January 23th, 2019.

    This initiative comes along with the CRS initiative to award young scientists with travel grants to those local chapters applying for those awards. These travel grants will enable students to participate in the 2019 CRS annual meeting (July 21-24, 2019, Valencia, Spain). Four travel grants will be awarded to the best oral and poster communications. The program includes 2 invited talks by Marta Vives-Pi (UniversitatAutònoma de Barcelona) and João F. Mano (University of Aveiro), well known leaders in their field of research, 15 oral presentations and 33 posters.

    In advance, we again thank you for your interest in 2nd SPLC-CRS Young Scientists Meeting.

    We look forward to meeting you in Santiago!

    On behalf of the Board of SPLC-CRS,

    Dolores Torres President of the SPLC-CRS

    María J. Blanco-Prieto Secretary of the SPLC-CRS

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    6

    Scientific Program

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    7

    Novoa Santos Auditorium, School of Medicine (USC)

    23/01/2019

    8:30 Registration

    Presentations upload/posters mounting

    8.50 Welcome

    Dolores Torres (USC; SPLC-CRS President)

    Maria José Alonso (USC; CRS President)

    9:00 Invited speaker – Marta Vives-Pi “Targeting autoimmune diseases with nanomedicine: The immune re-

    education”

    Head of the Immunology of Diabetes UnitGermans Trias i Pujol Research Institute (IGTP), Autonomous University of Barcelona

    Chairs: María Jesús Vicent (CIPF, Valencia) Bruno Sarmento (i3S; U Porto)

    Young Scientists Session I

    9:30 Polymeric nanocapsules as a novel tolerogenic treatment for Type 1 Diabetes - Ana Olivera Center for Research in Molecular Medicine and Chronic Diseases (CIMUS, University of Santiago de Compostela

    9:39 Development of in vitro dissolution tools predictive of the in vivo behaviour of pharmaceutical formulations: Candesartan in vitro-in vivo correlation (IVIVC) - Bárbara Sánchez Dengra Pharmacokinetics and Pharmaceutical Technology Area. Miguel Hernandez University

    9:48 Engineered albumin conjugated nanoparticles are a promising strategy to enhance FcRn-mediated delivery across intestinal barriers – Claudia Azevedo i3S Instituto de Investigação e Inovação em Saúde. Universidade do Porto

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    8

    9:57 Nanoencapsulated Bevacizumab Inhibits Glioblastoma Vascularization via Intratumoral VEGF Trapping – Flávia Sousa i3S Instituto de Investigação e Inovação em Saúde. Universidade do Porto

    10.06 A nanotechnological approach to improve the therapeutic efficacy of cytotoxic agents against melanoma – Jacinta Pinho Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa

    10:15 Evaluation of the immunotoxicological properties of PLA nanoparticles– Jessica da Silva Faculty of Pharmacy. University of Coimbra

    10:24 Casein nanoparticles as effective vehicles for enhancing the oral bioavailability of resveratrol – Jorge Morales Gracia Department of Chemistry and Pharmaceutical Technology, University of Navarra

    10:33 Design of an anti IGFR1 polymer-antibody conjugate in combination with Abiraterone for the treatment of Castration-Resistant Prostate Cancer (CRPC) – Katia Maso Polymer Therapeutics Lab, Centro de investigación Príncipe Felipe (CIPF), Valencia

    10:42 Combination of cardiomyocytes derived from human induced pluripotent stem cells with biomaterials restores cardiac function in infarcted mice – Laura Saludas Echauri Department of Chemistry and Pharmaceutical Technology, University of Navarra

    11.00 Coffee-break/ Posters Viewing

    11:30 Invited speaker – Joao Mano

    “Nano to macro- polymeric particles for the release of active molecules”

    Full Professor Department of Chemistry, CICECO - Aveiro Institute of Materials University of Aveiro, Portugal

    Chairs: Helena Florindo (U Lisboa) Manuel Santander (U Castilla-La Mancha)

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    9

    Young scientists session II

    12:00 Nanotechnology-based approach for the delivery of glycoconjugate vaccines - Maruthi Prasanna Center for Research in Molecular Medicine and Chronic Diseases (CIMUS, University of Santiago de Compostela

    12:09 Rational development and preclinical assessment of bio-inspired Protein-Metal Nanosystems as oral delivery carrier – Matilde Durán Lobato Center for Research in Molecular Medicine and Chronic Diseases (CIMUS, University of Santiago de Compostela

    12:18 Control of Loading and Polarized Orientation of Antibodies onto Gold Nanoparticles – Mireya López Borrajo Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), University of Santiago de Compostela, Spain

    12:27 Impact of PEG coating in the distribution and retention of PLGA nanoparticles in colorectal tissues – Rute Nunes I3S – Instituto de Investigação e Inovação em Saúde, Universidade do Porto

    12:36 Multimodal nanoemulsions for whole-body imaging in metastatic cancer – Sandra Diez VillaresNano-Oncology Unit, Translational Medical Oncology Group (ONCOMET). Health Research Institute of Santiago de Compostela (IDIS) and CIBERONC, Santiago de Compostela

    12.45 Combination of polymeric nanoparticles and alginate cryogel towards the development of an in situ cancer vaccine – Tomas Ramos i3S Instituto de Investigação e Inovação em Saúde e Instituto de Engenharia Biomédica, Universidade do Porto

    13:00 Award Ceremony for the best oral and poster presentations

    13:15 SPLC-CRS General Assembly

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    10

    Invited Speakers

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    11

    Targeting autoimmune diseases with nanomedicine: The

    immune re-education

    Marta Vives-Pi

    Head of the Immunology of Diabetes Unit

    Germans Trias i Pujol Research Institute (IGTP), Autonoma University, Barcelona

    Type 1 diabetes (T1D) is a metabolic disease caused by the autoimmune

    destruction of insulin-producing β-cells. With its incidence increasing worldwide,

    to find a safe approach to permanently cease autoimmunity and allow β-cell

    recovery has become vital. Relying on the inherent ability of apoptotic cells to

    induce immunological tolerance, we demonstrated that liposomes mimicking

    apoptotic β-cells arrested autoimmunity to β-cells and prevented experimental

    T1D through tolerogenic dendritic cell (DC) generation. This novel

    immunotherapy has the potential to re-establish immunological tolerance,

    opening the door to new therapeutic approaches in the field of autoimmunity.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    12

    Nano to macro- polymeric particles for the release of active molecules

    João F. Mano

    Department of Chemistry. CICECO. University of Aveiro. 3810-193 Aveiro. Portugal.

    Email: [email protected]

    Biodegradable and biocompatible polymeric particles have been widely used to

    encapsulate molecules that could then be released in a controlled or sustained

    way. This lecture will explore a range of technologies for the processing and

    application of particles for drug release ranging from the nano to the macro-scale

    sizes. Although the methodologies proposed are transversal for many

    pharmaceutical and bioengineering applications examples will especially focus

    the area of tissue engineering and regenerative medicine, in particular for bone

    repair. Nanoparticles can be used to deliver differentiation factors to direct the

    faith of stem cells. We discovered that naringin, a naturally occurring flavanone,

    encapsulated in nanomiceles could be internalised and released in mesenchymal

    stem cells, promoting the differentiation and augmenting the pro-osteogenic

    effect over that of free drug and standard induction methods, thus indicating the

    potential of this system to enhance stem cell–based bone regeneration.

    Micro and macro-particles should be prepared using other methodologies. We

    proposed the use of superhydrophobic substrates as platforms to produce

    hydrogel or polymeric particles able to encapsulate molecules with a high loading

    efficiency. Several examples are presented in this context using natural based

    polymers and synthetic biodegradable polymers. The mild conditions that the

    particles may be formed allows for the encapsulation of living cells, extending the

    applicability of the technology.

    Liquefied capsules are also an interesting system where drugs or cells may be

    encapsulated and released in a controllable fashion. We have been using the

    layer-by-layer technique (LbL) for the production of the multilayer shell that is

    enveloping the capsules. The mechanical and transport properties of the

    multilayers could be tuned, and the use of stimuli-responsive polymers in the

    construction of the shells permits to develop smarter delivery systems. The

    technology was also adapted to encapsulate living cells, and the resulting

    compartmentalized cell microfactories can be produced for distinct biomedical

    applications.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    13

    Abstracts: Oral presentations

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    ____________________________________________________________________________

    14

    O1. Polymeric nanocapsules as a novel tolerogenic treatment for Type 1 Diabetes

    Ana Olivera1,2, Cristina Calviño-Sampedro3, Rubén Varela-Calviño3, Dolores Torres2, María José

    Alonso1,2

    1Center for Research in Molecular Medicine and Chronic Diseases (CiMUS), Universidade de

    Santiago de Compostela, Spain 2Department of Pharmacology, Pharmacy and Pharmaceutical Technology, School of Pharmacy,

    Universidade de Santiago de Compostela, Spain 3Department of Biochemistry, School of Pharmacy, Universidade de Santiago de Compostela,

    Spain

    Type 1 diabetes (T1D) is considered a chronic autoimmune disease caused by the destruction of

    β-cells located in the Langerhans islets by the immune system, which leads to the loss of insulin

    production in pancreas. Within this context, specific treatments to overcome the immune reaction

    against β-cells are desirable. Nanotechnology offers the possibility of modulating the specific

    immune response, by developing a tolerogenic profile in the immune competent cells, thus

    avoiding the off-target effects1. In this report, we evaluated polymeric nanocapsules (NCs)

    containing a tolerogenic molecule as a therapy for Type 1 Diabetes. NCs were prepared using

    solvent-displacement technique, showing all the prototypes particle size around 200 nm and a

    surface charge around +50 mV. Upon 6 h of incubation of the NCs with human dendritic cells

    (hDCs), no signs of toxicity were observed up to a dose of 900 µg/mL. Fluorescence microscopy

    images showed the ability of the NCs to interact with hDCs. NCs were able to maintain the activity

    of the tolerogenic molecule and were effective at inducing a tolerogenic profile in hDCs. Finally,

    NCs were evaluated in NOD mice model, leading to a significant delay in the diabetes onset

    compared to controls.

    REFERENCES

    1. Dacoba T.G. et al., Semin Immunol. 34:78-102, 2017

    ACKNOWLEDGEMENTS

    Research reported in this publication was supported by the Spanish Ministry of Science,

    Innovation and Universities [grant agreement number 646142]. The first author is also supported

    by a FPI grant from the Spanish Ministry of Economy, Industry and Competitiveness (grant

    number BES-2015-071236).

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    15

    O2. Development of in vitro dissolution tools predictive of the in vivo behaviour of

    pharmaceutical formulations: Candesartan in vitro-in vivo correlation (IVIVC).

    Andrés Figueroa-Campos1,2, Bárbara Sánchez-Dengra1, Virginia Merino2, Isabel González-

    Álvarez1, Marta González-Álvarez1, Marival Bermejo1

    1Engineering: Pharmacokinetics and Pharmaceutical Technology Area. Miguel Hernandez

    University, San Juan de Alicante, Alicante, 03550, Spain. 2Pharmacy and Pharmaceutical Technology. Valencia University, Valencia, Valencia, 46100,

    Spain.

    Introduction

    An in vitro-in vivo correlation (IVIVC) is a mathematical relationship which, after being validated,

    is able to relate an in vitro characteristic of a drug (e.g. dissolution), with a characteristic of its

    biological behaviour in vivo (e.g. absorption). The objective of this work was to establish a level

    A IVIVC for three Candesartan oral immediate release formulations.

    Methods

    Plasma profiles of Candesartan for all the formulations were deconvoluted by Loo-Riegelman

    method to obtain the individual fractions absorbed (fa).

    Fractions dissolved (fdiss) were obtained in several conditions in USP II and USP IV apparatus

    and the most biopredictive in vitro experiment was chosen by comparison of dissolution

    profiles with the f2 similarity factor.

    Levy plot was constructed to estimate the time scaling factor and two two-steps IVIVCs (linear

    and polynomial) were obtained.

    The program Berkeley-Madonna was used for obtaining a one-step IVIVC.

    The percentages of prediction error (%PEs) were calculated as a predictability measure for

    validating the correlations.

    Results and discussion

    In vivo biological behaviour of Candesartan can be described by means of a two-compartment

    model. Dissolution profiles obtained in USP IV apparatus with media of pH 1.2, 4.5 and 6.8

    modified with 0.2% Tween 20 were selected as biopredictive for preparing the IVIVCs.

    Three level A IVIVCs were obtained, whose percentages of prediction error are lower than the

    pre-established limits (the individual %PE of each formulation for each parameter does not

    exceed 15% and the mean %PE of all formulations for each parameter is less than 10%), so they

    are valid and biopredictive.

    Conclusions

    All three IVIVCs could be used as substitutes of human bioequivalence studies, after their

    approval by the competent agencies. Nevertheless, the correlation considered more appropriate,

    at this moment, is the linear IVIVC obtained by the two-step method.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    ____________________________________________________________________________

    16

    O3. Engineered albumin conjugated nanoparticles are a promising strategy to

    enhance FcRn-mediated delivery across intestinal barriers

    C. Azevedo1,2,3, J. Nilsen4,5, A. Grevys4,6, R. Nunes1,2,3, J. T. Andersen4,6, B. Sarmento1,2,7

    1i3S – Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; 2Nanomedicines & Translational Drug Delivery Group, INEB - Instituto Nacional de Engenharia

    Biomédica, Portugal; 3Instituto de Ciências Biomédicas Abel Salazar, University of Porto, Portugal; 4Department of Immunology, Centre for Immune Regulation (CIR), Oslo University Hospital

    Rikshospitalet and University of Oslo, Oslo, Norway 5Institute of Clinical Medicine, University of Oslo, Oslo, Norway 6Department of Biosciences, University of Oslo, Norway. 7CESPU, Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde

    & Instituto Universitário de Ciências da Saúde, Gandra, Portugal

    To maintain patient compliance, oral administration is the preferred drug delivery route [1]. Yet,

    many drugs, including biopharmaceuticals do not endure the harsh environment of the

    gastrointestinal tract and cross epithelial barriers inefficiently. Functionalization of nanoparticles

    (NPs) with ligands that can bind specifically to surface receptors may be one strategy to overcome

    these obstacles and to improve the delivery of pharmaceuticals through biological barriers [2]. We

    aimed to explore whether polymeric nanoparticles (NPs) conjugated with engineered human

    albumin for enhanced FcRn binding [3, 4] could be an attractive strategy for delivery of

    encapsulated drugs across mucosal barriers. For this purpose, PLGA NPs were designed using

    double emulsion/evaporation technology where engineered human albumin was site-specifically

    conjugated to the polymers.

    NPs with approximately 150 nm, presented an insulin encapsulation efficiency over 80% and

    approximately 100% of the conjugated albumin on the surface. NPs-HSA variants bind with

    expected [5] binding hierarchy to hFcRn: NPs-KAHQ < NPs-WT < NPs-KP < NPs-TNNEKP. NPs-

    KP and NPs-TNNEKP present improved affinity for FcRn by 6- fold and 12-fold, respectively. This

    enhance FcRn-mediated delivery across intestinal barriers and contributes for the NPs extended

    half-life. This was confirmed by permeability studies and Human endothelial cell-based recycling

    assay (HERA), which suggest that NPs conjugated with high binder albumin variants are

    transported and recycled more efficiently, in an FcRn-dependent manner. The proof of concept

    was demonstrated in the preliminary pharmacodynamic study, using the state-of-the-art human

    FcRn transgenic mouse model. It was verified that NPs-TNNEKP present approximately 18% of

    relative hypoglycemic decrease and 6% of pharmacologic availability, after 24h of administration,

    when compared with insulin s.c. administered. Next, we will explore the pharmacokinetics as well

    as the biodistribution of the formulations.

    In short, the FcRn-targeted approach may pave the way for more efficient delivery of NP-

    encapsulated drugs.

    References

    [1] B. Sarmento, et al., L. Jorgensen and H.M. Nielsen, Editors, 2009, 207-227.

    [2] P. Fonte, et al, Biotechnology Advances, 2015, 33, 1342-1354.

    [3] D.C. Roopenian and S. Akilesh, Nature Reviews Immunology, 2007, 7, 715-725.

    [4] J. Martins, et al, Pharmacology & Therapeutics, 2016, 161, 22-39.

    [5] J.T. Andersen, et al., Journal of Biological Chemistry, 2014, 289(19): 13492-13502.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    17

    O4. Nanoencapsulated Bevacizumab Inhibits Glioblastoma Vascularization via

    Intratumoral VEGF Trapping

    Flávia Sousa1,2,3,4,5, Andrea Cruz5, Fábio Júnio Ferreira1,6, José Bessa1,6, Bruno Sarmento1,2,4, Inês Mendes Pinto5* 1i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen 208, 4200- 393 Porto, Portugal 2INEB - Instituto Nacional de Engenharia Biomédica, Universidade do Porto, Rua Alfredo Allen 208, 4200- 393 Porto, Portugal 3ICBAS - Instituto Ciências Biomédicas Abel Salazar, Universidade do Porto, 4150-180 Porto, Portugal 4CESPU - Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde, Rua Central de Gandra 1317, 4585-116 Gandra, Portugal 5INL, International Iberian Nanotechnology Laboratory, Avenida Mestre José Veiga, 4715-330 Braga, Portugal 6IBMC, Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua Alfredo Allen 208, 4200-393 Porto, Portugal

    Glioblastoma multiforme (GBM) is the most aggressive malignant brain tumor, being the median survival time of patients at around 15 months after disease diagnosis1. GBM has the constant need of vascularization, making this tumor one of the most vascularized and invasive solid tumors2. Bevacizumab, an anti-VEGF monoclonal antibody, was approved by FDA to be used as a single agent for patients with GBM3. Despite good results in the clinical trials, the low probability of bevacizumab in crossing the BBB limits its CNS accessibility and does not allow an improvement in the overall patient survival4. Therefore, an alternative to improve the efficacy of GBM treatment is highly needed and might be achieved by the combination of nanotechnology through controlled release nanosystems5. In this study, bevacizumab-loaded PLGA NP were successfully developed as an alternative to cross effectively BBB, accomplishing a better therapy6. No significant differences were also found by BrdU and ELISA assay for anti-proliferative and anti-VEGF properties between free and encapsulated bevacizumab, demonstrating the success of encapsulation. In vivo efficacy of bevacizumab-loaded PLGA NP was evaluated using a glioma zebrafish model to study the neoangiogenesis and tumor growth through the injection of GBM cancer cells. In vivo results showed a significant decrease in tumor area just for the bevacizumab-loaded PLGA NP group. Trying to understand the molecular mechanism behind the efficacy of nanoparticles, a cellular uptake in both cell lines was done to study the internalization of bevacizumab and its effect on VEGF secretion. A significant increase in the number of bevacizumab positive cells and a decrease in the number of VEGF producing cells was obtained for the bevacizumab-loaded PLGA NP group. These last results demonstrated that bevacizumab-loaded PLGA NP might cause a disorder in VEGF signaling pathway, being an efficient alternative to deliver intracellularly monoclonal antibodies.

    References 1 Ohgaki, H. Epidemiology of brain tumors. Methods Mol. Biol. 472, 323-342, doi:10.1007/978-1-

    60327-492-0_14 (2009). 2 Broekman, M. L. et al. Multidimensional communication in the microenvirons of glioblastoma. Nat. Rev. Neurol. 14, 482-495, doi:10.1038/s41582-018-0025-8 (2018).

    3 Cohen, M. H., Shen, Y. L., Keegan, P. & Pazdur, R. FDA drug approval summary: bevacizumab (Avastin) as treatment of recurrent glioblastoma multiforme. Oncologist 14, 1131-1138,

    doi:10.1634/theoncologist.2009-0121 (2009). 4 Thompson, E. M., Frenkel, E. P. & Neuwelt, E. A. The paradoxical effect of bevacizumab in the therapy of malignant gliomas. Neurology 76, 87-93, doi:10.1212/WNL.0b013e318204a3af (2011).

    5 Gomes, M. J., Fernandes, C., Martins, S., Borges, F. & Sarmento, B. Tailoring Lipid and Polymeric Nanoparticles as siRNA Carriers towards the Blood-Brain Barrier - from Targeting to Safe Administration. J. Neuroimmune Pharmacol. 12, 107-119, doi:10.1007/s11481-016-9685-6 (2017). 6 Sousa, F. et al. A new paradigm for antiangiogenic therapy through controlled release of bevacizumab from PLGA nanoparticles. Sci. Rep. 7, 3736, doi:10.1038/s41598-017-03959-4 (2017).

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    ____________________________________________________________________________

    18

    O5. A nanotechnological approach to improve the therapeutic efficacy of cytotoxic

    agents against melanoma

    Jacinta O. Pinho1, Eduarda Mendes1, Maria de Jesus Perry1, Joana D. Amaral1, Cecília M.P.

    Rodrigues1, Ana P. Francisco1, Angela Casini2, Graça Soveral1, M. Manuela Gaspar1

    1Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Lisbon, 1649-003, Portugal; 2School of Chemistry, Cardiff University, Cardiff, United Kingdom

    Melanoma is a major cause of morbidity and mortality, and effective therapeutic options are

    needed. Therefore, two distinct compounds have been explored by our research group. First, the

    copper complex Cuphen [1,2], recently reported as inhibitor of aquaporins (AQPs). These are a

    family of transmembrane proteins overexpressed in several malignancies and promising targets

    for cancer treatment [3]. Second, a newly synthesized triazene hybrid molecule (THM) [4]. THM

    displays two distinct moieties, acting by two mechanisms: 1) high specificity towards tyrosinase,

    an enzyme up-regulated in melanoma cells; 2) toxic alkylating properties.

    In vitro studies demonstrated the high cytotoxic effect of Cuphen and THM towards murine and

    human cancer cell lines, with IC50 values

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    19

    O6. Evaluation of the immunotoxicological properties of PLA nanoparticles

    Jessica Da Silva1;2, Sandra Jesus1;2, Natália Bernardi1;2 and Olga Borges1;2

    1Faculty of Pharmacy, University of Coimbra, Azinhaga de Santa Comba, 3000-548 Coimbra,

    Portugal 2Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, Portugal

    Polylactic acid (PLA) is a biodegradable, biocompatible and a Food and Drug Administration

    approved polymer, widely used as a platform for antigen and drug delivery, among others. [1, 2]

    Despite generally regarded as safe, the immunotoxicological profile of PLA, when used as a

    polymeric nanoparticle (NP), is not well documented. [3] Thus, this study intends to fill this gap, by

    evaluating the immunotoxicological profile of PLA NPs, and to compare it with other NPs such as

    polycaprolactone (PCL) and PCL/chitosan (PCL/Chi) NPs.

    All nanoparticles were produced by a nanoprecipitation method and were characterized regarding

    its mean diameter, polydispersity index and zeta potential. Then, several assays were performed

    in order to evaluate the immunotoxicological properties of each NP: cell viability, nitric oxide (NO)

    production and inhibition, and reactive oxygen species (ROS) production were evaluated in RAW

    264.7 cells; cell viability was assessed in human peripheral blood mononuclear cells (PBMCs);

    and hemolysis was studied in whole human blood.

    Two different size PLA NPs were produced (200 nm and 100 nm), as well as PCL NPs with ~170

    nm. PCL/Chi NPs presented ~270 nm and a distinctive positive zeta potential. Cell viability studies

    in RAW 264.7 showed higher toxicity for PCL/Chi NPs, with an IC50 of 222 µg/mL. Regarding

    ROS production, no effect was observed with the PCL and the PCL/Chi NPs under the

    concentration range tested (8.6-689.4 µg/mL). Interestingly, PLA NPs with larger size induced a

    dose-dependent ROS production. Also, none of the NPs under test induced NO production or

    inhibition (1-100 µg/mL), all induced cell viabilities in PBMCs above 70 % (1.1-1125 µg/mL) and

    presented a good hemocompatibility profile.

    Therefore, this study emphasizes the importance of establishing the immunotoxicological profile

    of different NPs, as we verified that differences in size, composition and zeta potential can

    influence the delivery system biological activity.

    [1] ESSA, S. et al. Improved antifungal activity of itraconazole-loaded PEG / PLA nanoparticles. v.

    30, n. 3, p. 205–217, 2013. [2] LEGAZ, S. et al. ST AC. v. 5390, n. April, 2016. [3] SINGH, R. P.; RAMARAO, P. Accumulated Polymer Degradation Products as Effector

    Molecules in Cytotoxicity of Polymeric Nanoparticles. v. 136, n. 1, p. 131–143, 2013.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    ____________________________________________________________________________

    20

    O7. Casein nanoparticles as effective vehicles for enhancing the oral

    bioavailability of resveratrol

    Rebeca Peñalva1, Jorge Morales1, Carlos J. González-Navarro2, Juan M. Irache1

    1Department of Chemistry and Pharmaceutical Technology, University of Navarra, 31008

    Pamplona, Spain. 2Centre for Nutrition Research, University of Navarra, 31080 Pamplona, Spain.

    Resveratrol is a naturally occurring polyphenol that provides several health benefits including

    cardioprotection and cancer prevention. However, its biological activity is limited by a poor

    bioavailability when taken orally. In addition, after the oral administration of resveratrol, it suffers

    a presystemic metabolism, which leads to the formation of a metabolite (resveratrol-3-O-

    glucuronide) that also results in a low oral bioavailability. The aim of this work was to evaluate the

    capability of casein nanoparticles as oral carriers for resveratrol. Nanoparticles were prepared by

    a coacervation process, purified and dried by spray-drying. The mean size of the resveratrol

    loaded nanoparticles was around 200 nm, with a payload close to 30 µg/mg nanoparticle.

    Resveratrol release from casein nanoparticles was not affected by the pH conditions and followed

    a zero-order kinetic. When nanoparticles were administered orally to rats, they remained within

    the gut, displaying an important capability to reach the intestinal epithelium. No evidence of

    nanoparticle “translocation” were observed. The resveratrol plasma levels were high and

    sustained for at least 8 hours, it was also quantified in plasma after 24 hours. Figure 1 shows the

    plasma concentration levels of resveratrol as a function of time after a single oral administration

    of 15 mg/kg to male Wistar rats of the different formulations tested (solution and casein

    nanoparticles). The oral bioavailability of resveratrol when loaded in casein nanoparticles was

    calculated to be 26.5%, ten-fold higher than when the polyphenol was administered as oral

    solution. The main metabolite of the polyphenol, resveratrol-3-O-glucuronide, was quantified in

    plasma after 24 hours. Finally, a good correlation between in vitro and in vivo data was observed.

    Figure 1. Resveratrol plasma concentration vs. time after a single oral administration of 15 mg/kg

    i) resveratrol PEG400:water solution (●), ii) resveratrol-loaded casein nanoparticles (■). Data

    expressed as mean± SD, (n= 6).

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    21

    O8. Design of an anti IGFR1 polymer-antibody conjugate in combination with

    Abiraterone for the treatment of Castration-Resistant Prostate Cancer (CRPC)

    Sonia Vicente-Ruiz1, Katia Maso1, Ana Armiñan1, Elena Gallon1, Julie Movellan1, David

    Charbonnier1, Fernanda Rodríguez1, María García-Flores2, José A. López-Guerrero2, María J.

    Vicent1

    1Polymer Therapeutics Lab, Centro de investigación Príncipe Felipe (CIPF), Valencia, Spain 2Fundación Instituto Valenciano de Oncología, Valencia, Spain

    A chromosomal rearrangement between TMPRSS2 (androgen-dependent serine protease) and

    ERG (transcription factor belonging to the ETS family) genes, that leds to the overexpression of

    TMPRSS2-ERG (T2E) transcript, is present in 50-70% of prostate cancer (PCa) patients.1,2 It has

    been showed that the combination of an inhibitor of type 1 insulin-like growth factor receptor

    (IGF1R) with an anti-androgen drug (Abiraterone) results in synergistic anti-tumor effects in T2E

    positive cells.3 In the present work, we have investigated the anti tumoral activity of a human

    monoclonal antibody (mAb) targeting IGF1R and its synergism with the anti-androgen drug

    against aggressive PCa. In particular, the anti IGFR1 mAb was covalently conjugated to poly-L-

    glutamic acid (PGA) through a reducible disulfide bridge and the polymer conjugate was fully

    characterized by GF chromatography, SDS-PAGE, DLS, FUV-CD and amino acid analysis. In

    vitro citotoxicity of mAb-PGA was evaluated in a panel of PCa cell lines (VCaP, LNCaP, PC3,

    22RV1, DU145 and RWPE1) with the MTS assay. However, only the VCaP cells, which express

    the T2E fusion gene, responded to both conjugated and unconjugated mAb. In addition, in vitro

    results disclosed that mAb-PGA in combination with Abiraterone displayed enhanced selectivity

    for VCaP cells respect to the combination with the unconjugated mAb. To further investigate this

    result, the compounds were Cy5.5 labelled and their intracellular fate in VCaP cells was followed

    by confocal and STORM microscopy, showing a different cellular trafficking. Moreover, to

    determine the cellular pathways related with treatments, we checked the differential proteins

    expression by Western Blot assay. We optimized an orthotopic PCa mice model employing

    luciferase-expressing VCaP cells and we iv injected unconjugated and conjugated mAb at the

    same dose for comparison. The results showed an higher mAb-PGA antitumoral activity respect

    to mAb. To corroborate the in vitro synergistic effects, we are currently testing the activity of the

    combination therapy with abiraterone in our in vivo model. So far, these results suggest that the

    combination anti IGFR1 mAb-PGA + Abiraterone may represent a promising therapeutic strategy

    for the T2E PCa patient subtype.

    Acknowledgements: This study is funded by PROMETEO/2016/103, SOGUG, Plan Nacional

    I+D; SAF2013-44848-R and FPI grant BES-2014-068439.

    References:

    1. Barbieri, CE et al. (2012) Molecular genetics of prostate cancer: emerging appreciation of genetic

    complexity. Histopathology. 60(1):187-98.

    2. Tomlins, SA et al. (2009) ETS gene fusions in prostate cancer: from discovery to daily clinical practice.

    European urology. 56(2):275-86.

    3. Mancarella, C et al. (2015) ERG deregulation induces IGF-1R expression in prostate cancer cells and

    effects sensitivity to anti-IGF-1R. Oncotarget. 30;6(18):16611-22.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    ____________________________________________________________________________

    22

    O9. Combination of cardiomyocytes derived from human induced pluripotent stem

    cells with biomaterials restores cardiac function in infarcted mice

    Laura Saludas1,2, Elisa Garbayo1,2, Manuel Mazo2,3, Beatriz Pelacho2,3, Gloria Abizanda2,3, Felipe

    Prósper2,3, María Blanco-Prieto1,2

    1 Department of Pharmaceutical Technology and Chemistry, Faculty of Pharmacy and Nutrition, University

    of Navarra, Pamplona, Spain 2 Instituto de Investigación Sanitaria de Navarra, IdiSNA, Pamplona, Spain 3 Area of Cell Therapy, Center for Applied Medical Research and Clínica Universidad de Navarra, Pamplona,

    Spain

    Cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) represent the only cell source with potential to remuscularize the infarcted area of the human heart [1]. Furthermore, they offer several other advantages such as proven effective paracrine effect, integration in the host myocardium and non-immunogenicity [2,3]. However, delivery issues related to the low cell survival and engraftment in the heart have limited the clinical implementation of an effective cell-based therapy for cardiac repair [4,5]. In this study, we proposed that the combination of hiPSC-CMs with microparticles (MPs) could constitute an effective strategy to improve the long-term engraftment of cells and therefore, to enhance their therapeutic effect. A homogenous population of poly(lactic-co-glycolic) acid MPs with a mean size of 10.4 ± 0.8 µm was obtained by the multiple emulsion solvent evaporation method. Next, particles were covered with collagen and poly-D-lysine to provide particles with a favourable biomimetic microenvironment for cell adhesion. CMs were obtained by differentiation of hiPSC by small-molecule manipulation of the Wnt-pathway, purified and adhered to the biomimetic MPs. First, the potential of particles to support cell survival was analyzed in vitro by the Alamar Blue assay, revealing a 1.99-fold increase in cell viability. Then, hiPSC-CM-MPs complexes were administered in a mouse myocardial infarction model. Interestingly, transplantation of hiPSC-CMs adhered to MPs improved the long-term engraftment of these cells up to two months. Furthermore, engrafted cells expressed dystrophin and connexin 43, which suggest that hiPSC-CMs maintained a cardiac phenotype and showed efficient electrical coupling through gap junctions. Finally, the improvement in the long-term retention of cells correlated with an increased heart function recovery. These findings suggest that MPs represent excellent platforms for cell delivery applications as they improve the long-term retention of cells. This strategy could be implemented also in other fields of regenerative medicine.

    References

    [1] Y.-W. Liu, B. Chen, X. Yang, J.A. et al., Human embryonic stem cell–derived cardiomyocytes

    restore function in infarcted hearts of non-human primates, Nat. Biotechnol. 36 (2018) 597–605.

    [2] D.A. Pijnappels, S. Gregoire, S.M. Wu, The integrative aspects of cardiac physiology and their

    implications for cell-based therapy., Ann. N. Y. Acad. Sci. 1188 (2010) 7–14.

    [3] A. Tachibana, M.R. Santoso, M. Mahmoudi, et al., Paracrine Effects of the Pluripotent Stem Cell-

    Derived Cardiac Myocytes Salvage the Injured Myocardium., Circ. Res. 121 (2017) e22–e36.

    [4] O. Iglesias-García, S. Baumgartner, L. Macrí-Pellizzeri, et al., Neuregulin-1β induces mature

    ventricular cardiac differentiation from induced pluripotent stem cells contributing to cardiac tissue

    repair., Stem Cells Dev. 24 (2015) 484–96.

    [5] J. V Terrovitis, R.R. Smith, E. Marbán, Assessment and optimization of cell engraftment after

    transplantation into the heart., Circ. Res. 106 (2010) 479–94.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    23

    O10. Nanotechnology-based approach for the delivery of glycoconjugate

    vaccines

    Maruthi Prasanna1,2, Aline Pillot2, Emilie Camberlein2, Cristina Calviño3, Ruben Varela3,

    Daphnée Soulard4, François Trottein4, Marcos Garcia-Fuentes1, Cyrille Grandjean2, Noemi

    Csaba1

    1Center for Research in Molecular Medicine and Chronic Diseases, University of Santiago de Compostela,

    Spain; 2Unit Function & Protein Engineering, UMR CNRS 6286, University of Nantes, France; 3Department of Biochemistry and Molecular Biology, University of Santiago de Compostela, Spain; 4Centre d’Infection et d’Immunité de Lille, Université de Lille, CHU Lille- Institut Pasteur de Lille, France

    The mortality of pneumonia has been greatly reduced by conjugate vaccines. However, their coverage is still limited due to serotype replacement. To overcome this, a glycoconjugate vaccine based on (pneumococcal surface adhesin-A (mPsaA)) is prepared. mPsaA is a protein antigen present in the surface of all pneumococcal serotypes. A synthetic mimic of pneumococcal serotype 14 tetrasaccharide (Pn14PS) was synthesized chemically and mPsaA was expressed in E. coli BL21 (DE3) strain. The obtained mPsaA and Pn14PS were successfully conjugated by thiol-maleimide coupling chemistry to obtain protein/sugar conjugates at ratio 1/5.4. The secondary structure of mPsaA was evaluated by circular dichroism after conjugation and lyophilization. The mPsaA-Pn14TS conjugate was loaded into chitosan nanoparticles (CNPs) prepared by an ionic gelation method. CNPs had a size of 118 ±3 nm and zeta potential of 31 ±1 mV; the encapsulation efficiency of the conjugate in CNPs was 70 ±3 %. Nanoparticle tracking analysis confirmed that CNPs had a particle count of 1.69 x 1010 per mg. The studies performed by scanning electron microscopy revealed that CNPs were spherical and had smooth surface. The CNPs were stable over a period of 24 h in simulated nasal fluid. The studies performed with human monocyte-derived dendritic cells (MoDCs) confirm the interaction of CNPs with antigen presenting cells. The internalization of fluorescently labelled CNPs by the MoDCs was confirmed by confocal microscopy and flow cytometry. The flow cytometry results showed the maximum uptake of nanoparticles in MoDCs was in less than 2 h. Immunization studies performed in a mouse model revealed that the groups immunized (S.C.) with CNPs produced a significantly higher IgG response against both Pn14TS and mPsaA, when compared to the mice immunized (S.C.) with mPsaA-Pn14TS alone. These results demonstrate the effectiveness of CNPs in generating enhanced immune response against both protein and carbohydrate antigens.

    References

    [1] Safari D et al., Nanomedicine (Lond) (2012) 7:651–662. [2] Larentis AL, et al., Protein Expr Purif (2011) 78:38–47. [3] Calvo P et al., Pharm Res (1997) 14:1431–1436.

    Acknowledgements The project has received funding from NanoFar, Erasmus Mundus Joint Doctorate in Nanomedicine and Pharmaceutical Innovation under European Union’s Horizon 2020 and RETOS-Spanish Ministry of Economy and Competitiveness (SAF2016-79230-R).

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    ____________________________________________________________________________

    24

    O11. Rational development and preclinical assessment of bio-inspired Protein-

    Metal Nanosystems as oral delivery carrier

    Matilde Durán-Lobato1, Juan Cunarro1, Sulay Tovar 1, Carlos Diéguez1,María J. Alonso1,2,3

    1Center for Research in Molecular Medicine and Chronic Diseases University of Santiago de

    Compostela, Spain 2IDIS Research Institute, Santiago de Compostela, Spain 3School of Pharmacy, University of Santiago de Compostela, Spain

    In spite of the high relevance of protein/peptide therapeutics in the current healthcare market scenario, the path from bench to beside still remains elusive for an oral peptide product. In order to design an oral protein nanocarrier product, aside from increasing oral bioavailability, other key features frequently sidelined should be considered, such as biocompatibility of materials and methods, reproducibility and scalability of the processes and the possibility of obtaining a final dosage form. Aiming at confronting these challenges, we explored the design of protein-metal nanosystems (Me-NSs) inspired on physiological materials and produced with industry-transferable methodologies. These Me-NSs were produced using an aqueous-based precipitation technique displaying a nanometer size below 100 nm with neutral ZP, association efficiency close to 100% and 10% protein loading. Upon contact with Simulated Intestinal Fluid (SIF), they proved to be colloidally stable while controlling protein release and protecting the protein cargo against enzymatic degradation in the presence of pancreatin. Furthermore, the production was successfully scaled-up by 10- and 100-fold increase in batch size and freeze-dried, leading to a powder product that maintained the physicochemical properties of the original formulation after mid-term RT storage. Moreover, the bioactivity of the associated protein was confirmed in an in vivo study, while the oral administration of the nanocarriers to rats proved their ability to enhance protein absorption. Overall, the results demonstrated the potential of Me-NSs as a bio-inspired and effective oral protein/peptide nanocarrier aiming at industrial translation.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    25

    O12. Control of Loading and Polarized Orientation of Antibodies onto Gold

    Nanoparticles

    Mireya L. Borrajo1, Víctor Puntes1,2,3

    1Vall d’Hebron Institut de Recerca (VHIR), 08035, Barcelona, Spain 2Institut Català de Nanociència i Nanotecnologia (ICN2), CSIC and The Barcelona Institut of

    Science and Technology (BIST), Campus UAB, 08193, Bellaterra, Barcelona, Spain 3Institució Catalana de Recerca i Estudis Avançats (ICREA), P. Lluís Companys, 23, 08010,

    Barcelona, Spain

    After the administration of inorganic nanoparticles (NPs), including gold nanoparticles (Au NPs),

    or their dispersion into biological fluids, the exposure to physiological proteins normally withstands

    the Protein Corona formation around the NP. The control over which proteins are present on this

    corona, their potential conformations and their absorption kinetics have been studied intensively

    in the past few years. Among them, antibodies have been conjugated to NP, showing a combined

    effect of the properties of both. The possibility of surface functionalization using more than one

    antibody can suppose the development of biomaterials with multiple functions (e.g. multiplexed

    diagnostics or multicell therapy). To achieve that, different conjugation strategies have been

    purposed; however, most of them implied complex and time-consuming chemistry in order to not

    only conjugate the antibodies onto the Au NPs, but also to maintain their functionality. Simple

    conjugation procedures are based on the hydrophobic and electrostatic interactions between

    antibodies and Au NPs, leading often to a poor functionality owing to the uncontrolled binding of

    the Fc and Fab regions of the antibodies to the NPs.

    Here we show the polarized conjugation of two different antibodies onto Au NPs, based on the

    preferential interaction of the Fc with the metallic surface, the spontaneous island growth of

    antibodies and their consequent segregation into different antibody domains. To characterize this

    Janus polarization, recognition antigen-antibody has been used to attach size different Au NPs.

    As it is shown in Fig. 1, Janus Au NPs (60 nm, functionalized with IgG anti-HSA and anti-BSA)

    interacts with Au NPs of 25 nm functionalized with HSA and Au NPs of 15 nm conjugated with

    BSA, demonstrating the polarized antibody distribution in the Au NP surface.

    Figure 1. (A) Schematic representation of the polarization of the antibodies onto the surface of the Au NP.

    (B) TEM characterization of the Janus distribution of the antibodies.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    ____________________________________________________________________________

    26

    O13. Impact of PEG coating in the distribution and retention of PLGA

    nanoparticles in colorectal tissues

    Rute Nunes1,2,3, Francisca Araújo1,2, Bruno Sarmento1,2,4, José das Neves1,2,4

    1I3S – Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal 2INEB - Instituto de Engenharia Biomédica, Universidade do Porto, Porto, Portugal 3ICBAS – Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, Porto,

    Portugal 4CESPU, Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da

    Saúde, Gandra, Portugal

    Mucus penetrating nanoparticles (NPs) obtained by dense polyethylene glycol (PEG) coating may

    be a powerful tool for the delivery of drugs in mucosal surfaces due to their ability to transpose

    the mucus barrier more efficiently than mucoadhesive ones. In this work, poly (lactic-co-glycolic

    acid) (PLGA NPs) were developed and non-covalently surface modified with poloxamer 407 – a

    triblock copolymer comprising polyethylene glycol and polypropylene glycol chains. PLGA NPs

    and PEG-PLGA presented sizes around 200 nm and narrow size distribution. The differences in

    zeta potential values for PLGA NPs and PEG-PLGA NPs were well-matched with the presence

    or not of PEG chains at NPs surface. The success of surface modification was also confirmed by 1H NMR, energy dispersive spectroscopy and contact angle. Developed NPs labeled with a near

    infrared probe were rectally administered to mice and the distribution and retention patterns in

    colorectal tissues were assessed by in vivo imaging. PEG-PLGA NPs were able to migrate farther

    into the colon and provide higher extension of coverage when compared to PLGA NPs after as

    little as 15 minutes of administration. PEG-PLGA NPs were able to cover around 40% of the total

    length of the colon, while PLGA NPs were restricted to less than 10%. Also, PEG-PLGA NPs

    showed higher residence in colorectal tissues between 15 min and 2 hours post-administration,

    as assessed by the analysis of the total fluorescence. Histological analysis of colorectal tissues

    of mice after rectal daily administration of PLGA NPs and PEG-PLGA NPs over 14 days revealed

    no architectural changes or cells infiltrates. Also, no differences in TNF-α, IFN-γ, IL-6 and IL1-β

    levels were observed between NPs and PBS treated animals. Overall, our results point out that

    PEG-PLGA NPs may be a useful platform for the mucosal delivery of drugs.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    27

    O14. Multimodal nanoemulsions for whole-body imaging in metastatic cancer.

    Sandra Díez-Villares1, Juan Pellico2, Ramón Iglesias-Rey3, Santiago Grijalvo4, Miguel A.

    Ramos-Docampo5, Verónica Salgueiriño5, Ramón Eritja4, Francisco Campos3, Fernando

    Herranz2, Rafael López-López1, María de la Fuente1.

    1Nano-Oncology Unit, Translational Medical Oncology Group (ONCOMET). Health Research

    Institute of Santiago de Compostela (IDIS) and CIBERONC, Santiago de Compostela, Spain. 2Nanomedicine and Radiochemistry Group, National Centre for Cardiovascular Research Carlos

    III (CNIC) and CIBERES, Madrid, Spain. 3Clinical Neurosciences Research Laboratory, Health Research Institute of Santiago de

    Compostela (IDIS), Santiago de Compostela, Spain. 4Nucleic Acids Chemistry Group, Institute of Advanced Chemistry of Catalonia (IQAC – CSIC)

    and CIBER-BBN, Barcelona, Spain. 5Magnetic Materials Group, University of Vigo, Vigo, Spain.

    Early diagnosis of metastatic cancer by whole-body techniques with high resolution and sensibility

    such as Magnetic Resonance Imaging (MRI) and Positron Emission Tomography (PET) could

    improve the outcome of patients and increase their survival. On view of this, we aimed to develop

    biocompatible nanoemulsions (NEs) that can incorporate MRI and PET probes and specifically

    reach disseminated cancer cells.

    NEs composed by sphingomyelin and oleic acid, were prepared by ethanol injection and

    characterized using a Nanosizer 2000®. NEs had a small colloidal size (130 nm), a monodisperse

    population (0.2), and a negative surface charge (-50 mV). Hydrophobic Superparamagnetic Iron

    Oxide Nanoparticles (SPIONs) coated with oleic acid were successfully incorporated into the oily

    nucleus of the NEs. These magnetic formulations were evaluated by MRI showing an excellent

    negative contrast (in vitro and in vivo) due to the Fe content and a high transverse relaxivity. For

    PET purposes, we synthesized a stearylamine derivative of the chelating agent NOTA (NOTA-

    SA) and it was efficiently associated to the NEs, without modifying their physicochemical

    properties. Afterwards, 68Ga was incubated with the NEs and purified by ultrafiltration, obtaining

    a high percentage of radioisotope association (86%) and a good stability in mice serum. NEs were

    injected intravenously in healthy mice, and PET/CT images were acquired at 2h post-

    administration. Biodistribution studies were done by the collection of mice organs and the

    quantification of their radioactivity at the end of the experiment. Results showed a good signal of

    the PET probe, not cardiotoxicity and a high detoxification by the liver, kidneys and spleen.

    In conclusion, we have developed biocompatible nanoemulsions that have potential for the

    development of novel tools for MRI and PET diagnosis. Next experiments will be aimed to explore

    their potential in a mice model of metastatic disease.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    ____________________________________________________________________________

    28

    O15. Combination of polymeric nanoparticles and alginate cryogel towards the development of an in situ cancer vaccine

    Tomás Bauleth-Ramos1,2,3,4,5,6, Ting-Yu Shih5,6, Mohammad-Ali Shahbazi4,7, Alexander J. Najibi5,6, Angelo S. Mao5,6, Pedro Granja1,2,3, Hélder A. Santos4,8, Bruno Sarmento1,2,9, David J. Mooney5,6 1 Instituto de Investigação e Inovação em Saúde (I3S), University of Porto, Rua Alfredo Allen, 208, 4200-135, Porto, Portugal 2 Instituto de Engenharia Biomédica (INEB), University of Porto, Rua Alfredo Allen, 208, 4200-135, Porto, Portugal 3 Instituto Ciências Biomédicas Abel Salazar (ICBAS), University of Porto, Rua Jorge Viterbo 228, 4150-180, Porto, Portugal 4 Drug Research Program, Division of Pharmaceutical Chemistry and Technology, Faculty of Pharmacy, University of Helsinki, FI-00014, Helsinki, Finland 5 John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA, 02138, USA 6 Wyss Institute Biologically Inspired Engineering, Harvard University, Boston, MA, 02115, USA 7Department of Pharmaceutical Nanotechnology, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran 8Helsinki Institute of Life Science 9CESPU, Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde, Gandra, Portugal

    INTRODUCTION Immunotherapies for cancer had an outstanding development in the past decade. However, the lack of antigen accumulation and proper stimulation of the immune cells led to poor antitumor responses.[1] Biomaterials have shown promising results as tools to surpass these issues.[2] Considering this, we hypothesized that by combining nutlin-3a (Nut3a) loaded nanoparticles (NPs) within an alginate cryogel loaded with granulocyte-macrophage colony-stimulating factor (GM-CSF) and cytosine-phospodiester-guanine-oligonucleotide (CpG-ODN), it would be possible to enhance the delivery of the drug, increase the therapeutic efficacy and simultaneously modulate the immune cells, creating an in situ vaccine. METHODS Nut3a loaded spermine-modified acetalated dextran NPs were prepared by double emulsion[3] and GM-CSF and CpG-ODN loaded alginate cryogels by covalent and ionic crosslinking.[4] NPs were physically adsorbed onto the cryogel and the system characterized by DLS, TEM, confocal microscopy, HPLC and fluorometer. In vitro association studies, biocompatibility, toxicity and immunogenicity were done in EL4 cancer cells. Biodistribution and efficacy studies were performed in vivo in C57BL6 mice bearing EL4 tumors. RESULTS Nut3a encapsulated NPs with positive surface charge and homogenous size distribution (221±4 nm) were formed and successfully adsorbed into the cryogels (Figure 1A). The NPs suffered a burst release in the first 6h and controlled release up to 72h (figure1B). Furthermore, NPs loaded gels enhanced tumor accumulation compared to intravenous injection (Figure 1C). Drug loaded NPs exerted an anticancer effect in vitro (Figure 1D) and Nut3a induced immunogenic cell death and slowed the tumor growth and increased survival in vivo. CONCLUSION Our results show that the developed combinatorial system can promote an anticancer effect, induce immunogenic cell death and increase the accumulation of nanoparticles in the tumor site. We believe that by combining the advantages of the drug loaded NPs with the GM-CSF/CpG loaded cryogel scaffold, it will be possible to exert a strong anticancer effect.

    References

    1. Wang, H. and D.J. Mooney, Biomaterial-assisted targeted modulation of immune cells in cancer treatment. Nat Mater, 2018. 17(9): p. 761-772.

    2. Weber, J.S. and J.J. Mule, Cancer immunotherapy meets biomaterials. Nat Biotechnol, 2015. 33(1): p. 44-5.

    3. Bauleth-Ramos, T., et al., Nutlin-3a and Cytokine Co-loaded Spermine-Modified Acetalated Dextran Nanoparticles for Cancer Chemo-Immunotherapy. Advanced Functional Materials, 2017. 27(42).

    4. Shih, T.Y., et al., Injectable, Tough Alginate Cryogels as Cancer Vaccines. Advanced Healthcare Materials, 2018. 7(10).

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    ____________________________________________________________________________

    29

    Abstracts: Poster presentations

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    ____________________________________________________________________________

    30

    P1. Rational design of Next-Generation Exosomas for targeted drug delivery in

    cancer therapeutics.

    Abi Judit Vazquez-Rios1,2, Ángela Molina-Crespo3, Sandra Alijas-Pérez2, Belén L Bouzo1,2,

    Gema Moreno-Bueno3, María de la Fuente Freire1.

    1 University of Santiago de Compostela (USC), Spain. 2 Nano-Oncology Unit, Translational Medical Oncology Group, Health Research Institute of

    Santiago de Compostela (IDIS), SERGAS, CIBERONC, 15706 Santiago de Compostela, Spain. 3 Translational Cancer Research Laboratory, Department of Biochemistry, Autonomous

    University of Madrid, School of Medicine, "Alberto Sols" Biomedical Research Institute CSIC-

    UAM, Madrid, Spain.

    In the past decade, exosomes have been proposed as ideal drug delivery systems for a broad

    range of pathologies, including cancer, inflammatory and regenerative diseases. Recent reports

    have shown impressive results in vivo that have prompted them into early clinical studies.

    However, important limitations related to their cumbersome and costly production processes as

    well as safety concerns are hampering their translation to clinic. Making use of the well-known

    liposome technology we propose the engineering of Next-Generation Exosomes (NGE) with clear

    advantages over natural exosomes in terms of yield of production, safety, and cost-effectiveness.

    NGE show great similarities to natural exosomes with respect to their physicochemical

    properties, ability to interact with the target cells, and to deliver innovative anticancer

    therapeutics. Moreover, NGE have been successfully functionalized with membrane proteins with

    organotropic properties, and have demonstrated to efficiently transport bioactive macromolecules

    (microRNAs) to the target cancer cells, in a similar fashion to tumoral exosomes. We present in

    this work a complete characterization of the proposed nanoplatform and evidence of the its

    potential for the targeted delivery of therapeutic RNAs to cancer cells in vitro and in vivo in an

    orthotopic xenograft model of human lung adenocarcinoma.

    We provide the first proof-of-concept of the potential of this technological nanoplatform as a real

    alternative to exosomes for the development of safer and more efficient anticancer therapies, a

    technology that is versatile and can be adapted as we go deep in the study of exosomes and the

    molecular features related to their tumor-homing properties. NGE provide additional opportunities

    of being further amendable to surface functionalization as well as loading of therapeutic cargo for

    a broader application in other fields.

    Taken together, our results suggest that NGE gain competitive advantage for drug delivery of

    anticancer therapeutics and provide a real alternative to natural exosomes for a widespread

    translation.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    31

    P2. Assessment of immunotoxicological features of chitosan nanoparticles

    Alana Duarte1,2#, Ana Patrícia Marques1,2#, Mariana Colaço1,2, Sandra Jesus1,2, Olga Borges1,2

    1Center for Neuroscience and Cell Biology, University of Coimbra, Portugal.2Department of Pharmaceutical Technology, Faculty of Pharmacy, University of Coimbra, Portugal.

    # These authors contributed equally to this work

    Nanoparticles (NPs) conquered an important role in drug delivery. Nonetheless, their effects on the immune system is poorly understood. So, it is necessary to develop trials and guidelines for the immunotoxicological assessment of nanomaterials to develop safe-by-design (SbD) NPs, minimizing health risks. Chitosan (Chi) is a natural polymer, that has been explored as a drug delivery vehicle. However, its immunotoxicological evaluation show contradictory results. In several reports, different physicochemical characteristics of Chi (molecular weight (MW) or deacetylation degree (DD)), the NPs size or endotoxin contamination are not reported, and can influence the immunotoxicity of the delivery system. The aim of this work is to study the immunotoxicity of NPs produced with chitosans with different DD (80 % and 93 %) and MW, to establish methods for immune function evaluation for SbD NP development. Chi NPs were prepared using a coacervation method and its size and zeta potential were characterized. Studies of toxicity were performed in human peripheral blood mononuclear cells (PBMCs) for MTT assay, in RAW 264.7 cells for MTT assay, for reactive oxygen species (ROS) and nitric oxide (NO) production, and in whole blood for hemolysis and coagulation time assays. The Chi NPs 80 % showed to be more toxic than Chi NPs 93 % in PBMCs. Both NPs and polymers decreased LPS-induced NO production but showed no stimulation in NO production and caused no hemolysis. Only Chi NPs 80 % increased ROS production and had a significant effect on plasma coagulation times, affecting the intrinsic pathway.

    We concluded that the DD and MW of Chi in the NPs can influence the results of cytotoxicity, coagulation times and ROS. These results together with further studies will contribute to develop guidelines to implement the SbD approach for nanobiomaterials, with focus on polymeric drug delivery systems.

    This work is funded by FEDER funds through the Operational Programme Competitiveness

    Factors - COMPETE 2020 and national funds by FCT - Foundation for Science and Technology

    under the project PROSAFE/0001/2016 and strategic project POCI-01-0145-FEDER-007440.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    ____________________________________________________________________________

    32

    P3. Antibiotic and anti-inflammatory controlled delivery from intraocular lenses for

    prophylaxis of endophthalmitis

    Ana Topete1,2, Benilde Saramago1, Ana Paula Serro2

    1Centro de Química Estrutural, Instituto Superior Técnico, University of Lisbon, Lisbon, Portugal; 2Centro de Investigação Interdisciplinar Egas Moniz, Instituto Universitário Egas Moniz, Monte da

    Caparica, Portugal

    Abstract:

    Cataract is the leading cause of blindness, affecting more than 20 million people worldwide each

    year. Cataracts surgery involves the removal of the opacified natural lens and implantation of an

    intraocular lens (IOL). In the post-operatory period, complications such as endophthalmitis can

    appear. To prevent this, antibiotics and anti-inflammatories eye drops are administered with a

    frequent posology. This drug delivery method is not very effective and leads to low patient

    compliance. In this work, it is explored the possibility of using drug-loaded IOLs to ensure a

    simultaneous release of both antibiotic and anti-inflammatory drugs.

    An antibiotic, moxifloxacin (MXF), and an anti-inflammatory, ketorolac (KTL) were loaded in

    acrylic IOLs by soaking in drugs solutions at 60°C for 2 weeks. Loading was carried out in a

    double drug solution of MXF+KTL and sequentially, in solutions containing just one of the drugs

    for comparison purposes. The loaded lenses were sterilized by autoclaving (121ºC, 1bar, 1h). In

    vitro drug release experiments were carried out in sink conditions. The effect of the drug loading

    and of the sterilization on lenses properties such as the swelling capacity, optical properties

    (transmittance) and mechanical properties (Young’s modulus) were evaluated.

    It was observed that, the double loaded lenses released a significant higher amount of each drug

    than the single loaded lenses. The devices were able to release effective amounts of the drugs

    for 2-4 weeks. Sterilization did not affect the release. It was found that the presence of drugs

    increases the swelling capacity of the lenses. Concerning the optical and mechanical properties,

    the changes observed do not compromise the use of the lenses.

    The sterilized double loaded lenses seem promising devices for the post-cataract surgery

    prophylaxis, complying with both antibiotic and anti-inflammatory therapeutic needs.

    Acknowledgements:

    The authors acknowledge funding from Fundação para a Ciência e Tecnologia (FCT) [projects

    UID/QUI/00100/2013 and PTDC/CTM-BIO/3640/2014] and support from PhysIOL for providing

    the materials for IOLs.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    33

    P4. Chitosan modifications into new amphiphilic derivatives for polymeric

    micelles production as drug delivery systems

    Andreia Almeida1,2,3, Marco Araújo1,2, Ramon Novoa-Carballal4,5, Cristina C. Barrias1,2,3, Marlene

    Lúcio6, Bruno Sarmento1,2,7

    1I3S – Institute for Research and Innovation in Health, University of Porto, Portugal2INEB – Institute of Biomedical Engineering, University of Porto, Rua Alfredo Allen, 208, 4200-

    135 Porto, Portugal 3ICBAS – Institute of Biomedical Sciences Abel Salazar, University of Porto, Rua de Jorge Viterbo

    Ferreira 228, 4050-313 Porto, Portugal 43B’s Research Group − Biomaterials, Biodegradables and Biomimetics, University of Minho,

    Headquarters of the European Institute of Excellence in Tissue Engineering and Regenerative

    Medicine, AvePark, 4806-909 Taipas, Guimarães, Portugal 5ICVS/3B’s PT Government Associate Laboratory, Braga/Guimarães, Portugal 6CFUM – Centre of Physics of University of Minho and Porto, Campus de Gualtar, 4710-057

    Braga, Portugal 7CESPU, IINFACTS – Institute for Research and Advanced Training in Health Sciences and

    Technologies, Rua Central de Gandra 1317, 4585-116 Gandra, Portugal

    Chitosan is a biocompatible and biodegradable polymer extensively explored in the development

    of drug delivery systems. The production of chitosan derivatives with amphiphilic properties have

    been widely investigated for delivery of anticancer agents. Polymeric micelles can be produced

    using block copolymers composed of hydrophilic and hydrophobic polymers or using lipids-graft-

    hydrophilic polymers. These systems allow to encapsulate drugs with different polarities, but with

    higher stability and have been explored as solubility enhancers of many anticancer drugs. Despite

    the promising results, the bioavailability and pharmacokinetic profile of oral administered

    formulations can be compromised by a release and degradation of the drug at gastric environment

    or reduced retention at the intestine and colon. These drawbacks can be overcome by the

    development of mucoadhesive formulations that also protect and retain the drug at gastric pH.

    Our work proposes to address these inadequacies using nanotechnology to develop oral

    mucoadhesive polymeric micelles based on chitosan for delivery of anticancer drugs. Chitosan

    derivatives were synthesized by a carbodiimide reaction by amidation and the success of the

    reactions was confirmed by Fourier transform infrared spectroscopy (FTIR) and hydrogen nuclear

    magnetic resonance analysis (1H NMR). Micelles were produced by solvent evaporation method,

    and the critical micelle concentration was also investigated. The obtained micelles presented a

    mean particle size in the range of 200-300 nm and positive surface charge. The morphology of

    micelles assessed by transmission electron microscopy (TEM) revealed round and smooth

    surface, in agreement with dynamic light scattering measurements. The association efficiency

    was determined by high-performance liquid chromatography (HPLC) and the cytotoxicity studies

    against Caco-2 and HT29-MTX intestinal epithelial cells demonstrated absence of cell toxicity for

    all formulations. Moreover, the intestinal permeability in Caco-2 monolayer and Caco-2/HT29-

    MTX co-culture model was also determined where chitosan systems presented higher intestinal

    permeability compared with the free drug.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    ____________________________________________________________________________

    34

    P5. Monodisperse glycosaminoglycan-based microgels for in situ protein

    delivery from tissue-mimicking implants

    Anna Abbadessa1,2, Carl C. L. Schuurmans1,3, Mikkel A. Bengtson1, Galja Pletikapic4, Huseyin

    Burak Eral5,6, Gijsje Koenderink4, Rosalinde Masereeuw3, Wim E. Hennink1 and Tina Vermonden1

    1Department of Pharmaceutics, Utrecht University, Utrecht, The Netherlands 2Later post-doc at Department of Fibre and Polymer Technology, Royal Institute of Technology (KTH), Stockholm, Sweden 3Department of Pharmacology, Utrecht University, Utrecht, The Netherlands 4Biological Soft Matter Group, AMOLF, Amsterdam, The Netherlands 5Process and Energy Department, Delft University of Technology, Delft, The Netherlands 6Van’t Hoff Laboratory for Physical and Colloid Chemistry, Utrecht University, Utrecht, The Netherlands

    Biological augmentation, via e.g. growth factors, of tissue-mimicking implants is a fundamental

    aspect in regenerative medicine. In this study1, we have investigated the potential of

    glycosaminoglycans (GAGs) microgels as protein carriers, as well as their protein release kinetics

    upon incorporation into hydrogel scaffolds designed for articular cartilage regeneration.

    Monodisperse microgels (500-700 μm) based on methacrylated hyaluronic acid (HAMA) and

    chondroitin sulfate (CSMA) were fabricated using a microfluidic device. They were subsequently

    loaded with a cationic model protein, i.e. lysozyme (Lys), via complex coacervation under two

    different buffer conditions (20 and 170 mM ionic strength, pH 7.4). Protein up-take, distribution

    and release kinetics from the microgel matrices were studied. Composite hydrogels made of Lys-

    loaded CSMA microgels embedded into hydrogel scaffolds were investigated in terms of Lys

    release, and were compared to Lys-loaded, CSMA-free hydrogel scaffolds and CSMA/hydrogel

    blends. In 20 mM buffer a higher Lys loading (up to 82%) was found for CSMA microgels

    compared to HAMA (76%), reasonably due to the higher CSMA charge density. Unlike CSMA

    microgels, HAMA microgels showed a significant decrease in Lys loading (51%) in 170 mM

    buffer, likely caused by the charge shielding originating from the higher ion concentration. Lys

    was homogeneously distributed throughout HAMA and CSMA microgels. In 170 mM PBS buffer

    (pH 7.4), Lys was released slower from CSMA than from HAMA microgels (total release around

    day 28 and 14, respectively), likely due to the stronger Lys-CSMA electrostatic interaction.

    Composite hydrogels released fully active Lys within 58 days, which is a much longer time frame

    than that showed by CSMA-free hydrogels (3 days) and CSMA/hydrogel blends (41 days). This

    study demonstrates that GAG microsphere/hydrogel composites loaded with cationic proteins via

    complex coacervation, are promising for the in situ protein delivery from tissue-engineered

    constructs.

    References [1] Soft Matter, 2018, 14 (30) 6327-6341.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    35

    P6. Caffeic Acid skin absorption: delivery of microparticles to hair follicles

    Lia Carolina Oliveira dos Santos1, Antonio José Guillot 2, Caroline Magnani Spagnol1, Marcos Antonio Corrêa1, Ana Melero Zaera 2.

    1Department of Drugs and Medicines, São Paulo State University (UNESP), School pf Pharmaceutical Sciences, Araraquara, SP, Brasil. Rod. Araraquara Jau Km 1- Campus Ville- Araraquara, São Paulo, Brazil CEP:14800-903 - Araraquara, SP. Tel.: (16) 3301-6880 Fax: (16) 3322-0073.

    2Department of Pharmacy and Pharmaceutical Technology and Parasitology. University of Valencia. Avda. Vicente A. Estelles SN. 46100, Burjassot. Valencia, Spain

    Caffeic acid (CA) is a polyphenol that can be found in many vegetal dietary sources. CA has a

    remarkable antioxidant potential and its antimicrobial action has been demonstrated through in

    vitro studies. Emulsions, especially O / W, are commonly used topical vehicles for active

    substances, since they present several advantages over other dosage forms. The challenge of

    dermal application for a drug is to overcome the anatomical and physiological barrier and to

    provide the effective concentration of the compound at active site. Therefore, it is necessary to

    investigate alternative routes, such as the transfollicular one. The objective of this work was to

    investigate the potential of transfollicular delivery of a topical emulsion containing CA formulated

    in microparticles and also non-encapsulated for comparative reasons. The tape stripping

    technique is widely used and accepted to study the kinetics and penetration of active substances

    via the skin, allowing to determine the location and distribution of substances within the Stratum

    corneum (SC). The Differential Stripping method is the most straightforward technique to

    determine follicular uptake quantitatively. Emulsions containing free CA and microencapsulated

    CA were applied over a predetermined area in the outer side of porcine ear skin. After the

    predetermined incubation time under constant temperature conditions, the SC was removed by

    the tape-stripping technique. To analyze the extent of follicular drug penetration, cyanoacrylate

    glue was applied over the pretreated skin area followed by its subsequent removal from the skin

    surface. The results of the retention studies demonstrated that, compared to the free drug, the

    microparticles allow a more even distribution of CA through the SC. Furthermore, as they are

    retained in the hair follicle, the drug can be released over a prolonged time in the hair follicle,

    where it can diffuse through the SC, which is much thinner in this appendage. Furthermore, this

    route can be explored to locally target the hair follicles with CA for the treatment of acne and hair

    loss, due to its pharmacological activity.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    ____________________________________________________________________________

    36

    P7. Polyphosphazene nanocarriers for gene delivery in cancer

    Carla Garcia-Mazas, Noemi Csaba, Marcos Garcia-Fuentes

    Center for Research in Molecular Medicine and Chronic Disease (CiMUS) and Dept.

    Pharmacology, Pharmacy and Pharmaceutical Technology, University of Santiago de

    Compostela

    Gene therapy has enormous potential for the treatment of a large number of diseases, but is still

    limited by the need of an efficient vector to allow entry in the cells. While viral vectors typically

    offer higher efficacy, there are still concern on their safety. Non-viral viral vectors are quickly

    improving their efficacy and they provide a safer alternative. Polymeric nanocarries are widely

    used to control the release of different therapeutic agents, both in time and space, which, in turn

    enhances drug efficacy and safety. Particularly, synthetic polymers offer the possibility of tailoring

    their properties through rational design, and possibility to obtain well-defined structures.

    In this work, we have developed and characterized new polymeric nanoparticles for gene therapy

    based on polyphosphazenes (PPZ). Polyphosphazenes are characterized by an inorganic

    phosphorous/nitrogen backbone, and by two side chains that can be modulated. Some

    polyphosphazenes have been investigated previously for gene therapy showing remarkable

    activity/toxicity ratio. Herein, we used this platform material to synthesize a cationic homopolymer

    (HoCat), a anionic homopolymer (HoAn), and a random copolymer based on HoCat were part of

    the amine groups were substituted by neutral moieties. These polymers were characterized by

    NMR for their chemical structure and then were used to form polyplexes with pDNA alone or in

    combination. The resulting nanoparticles were characterized for their size, pDNA

    association/dissociation capacity, toxicity and transfection capacity.

    The results showed that the nanosystems showed small particle size (90-100nm) and the capacity

    to condense the genetic material and to release it in presence of a polyanionic competitor. Toxicity

    tests indicated that the association of cationic with anionic PPZs resulted in improved cell viability

    as compared to cationic polyplexes alone. The same tendency was observed for cell transfection,

    where polyplexes containing cationic and anionic polymers improved the transfection in

    comparison with the cationic polyplexes alone.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    _____________________________________________________________________________

    37

    P8. Alkyl-lysophospholipid edelfosine and squalenoyl-gemcitabine form a

    novel nanomedicine applicable to cancer therapy

    Carlos Rodríguez-Nogales1,2, Víctor Sebastián3,4, Silvia Irusta3,4, Didier Desmaële5, Patrick Couvreur5, María J. Blanco-Prieto1,2

    1Pharmacy and Pharmaceutical Technology Department, University of Navarra, Pamplona

    31008, Spain2Instituto de Investigación Sanitaria de Navarra (IdiSNA), Pamplona 31008, Spain 3Department of Chemical and Environmental Engineering & Institute of Nanoscience of Aragon

    (INA), University of Zaragoza, Zaragoza 50018, Spain 4Networking Research Center on Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN,

    Madrid 28029, Spain. 5Institut Galien Paris-Sud, UMR CNRS 8612, Université Paris-Sud, Université Paris-Saclay,

    Châtenay-Malabry Cedex 92290, France

    The bioconjugation of drugs to squalenic acid, a nontoxic and biocompatible lipid squalene-

    derived molecule, leads to the formation of squalenoyl prodrugs able to self-assemble in

    supramolecular structures to form stable nanocomposites [1,2]. Among them, the prodrug

    squalenoyl-gemcitabine has been chosen to formulate a novel multidrug nanocomposite in

    combination with edelfosine, an alkyl-lysophopholipid with proven anticancer activity [3,4]. The

    main objective is to study if the physical mixture of both compounds could lead to the formation

    of a new anticancer nanomedicine. Chemical synthesis of squalenoyl-gemcitabine was performed

    according to previous studies [5,6] and nanocomposites were formulated by nanoprecipitation

    method. The physico-chemical properties of the nanoassemblies and its efficacy in pediatric

    osteosarcoma cells were evaluated. Designed and optimized squalenoyl-gemcitabine/edelfosine

    nanoassemblies showed a final mean particle size of 51±1 nm, PDI of 0.12 ±0.03 and zeta

    potential of -12.56 ±1.15 mv. High angle annular dark field scanning transmission electron

    microscopy showed the multilamellar assembly of squalenoyl-gemcitabine/edelfosine

    nanoassemblies, displaying a concentric or spiral disposition of the layers. Hemolysis

    experiments demonstrated that the nanoassemblies protected erythrocytes from the inherent

    hemolysis caused by non-encapsulated edelfosine. Cell culture viability tests in U2-OS confirmed

    that the co-assembly of edelfosine and squalenoyl-gemcitabine was not detrimental to the

    inherent antitumor activity of squalenoyl-gemcitabine nanoassemblies. Moreover, the novel

    nanoformulation killed the remaining plateau cells that were non-responsive to higher doses of

    squalenoyl gemcitabine nanoassemblies. Compared to the previous formulation, the combination

    of squalenoyl gemcitabine with edelfosine has resulted in a smaller particle size with higher

    stability and drug content whereas its antitumoral potential has remained intact. All in all, this

    concept may represent a step ahead toward a feasible multidrug nanoassembly that embodies

    the properties of a new generation of only drug-based nanomedicines.

    References

    [1] L.H. Reddy, J.-M. Renoir et al., Mol. Pharm. 6 (2009) 1526–1535.

    [2] F. Dosio, L.H. Reddy et al,, Bioconjug. Chem. 21 (2010) 1349–1361.

    [3] Y. González-Fernández, E. Imbuluzqueta et al., Cancer Lett. 388 (2017) 262–268.

    [4] A. Estella-Hermoso de Mendoza, V. Préat et al., J. Control. Release. 156 (2011) 421–426.

    [5] L.H. Reddy, C. Dubernet et al., J. Control. Release. 124 (2007) 20–27.

  • 2nd SPLC-CRS Young Scientists Meeting, January 23rd, 2019

    Santiago de Compostela, Spain

    ____________________________________________________________________________