WP1. Design, fabrication & optimization of selenium nanosystems (R&T)
Define the composition of the different seleno-compounds cocktails and Selection of most favourable materials for the production of nanodevices under GMP with the available technology within the consortium. Definition of coupling chemistry depending on the nature of the chemical nature of the nanodevice. Characterization of the nanosystems. The cytotoxicity of different variations will be examined in breast cancer cell lines and control cell lines.
Speciation of selenium before (after loading nanosystems, drug release, stability) and after cell uptake . Measurement of Se will be carried out by two different approaches: 1) Indirect speciation methods: control of Se-compounds will be carried out before and after in vitro experiments by applying Se speciation protocols including sample pre-treatment, enzymatic digestion, Se species separation, identification and quantification by using HPLC-ICP-MS; and 2) Direct speciation methods, based on direct observation of samples by high precision X-rays spectroscopic methods using Synchrotron based techniques (XANES and EXAFS), taking advantage of both the UAB expertise and the actual collaboration with the various European Synchrotrons, in particular ALBA synchrotron located in the surroundings of UAB. By using these methods, we will be able not only to determine the Se species but also their distribution in the target cells. This information will be of key importance to improve and personalize clinical trials.
WP2 TARGETING OF NANOSYSTEMS
This work package involve the antibody selection and production, followed by the coupling of antibodies onto the surface of nanocapsules and purification. The intracellular uptake of antibody conjugated NPs will be characterised by checking fluorescence labelled NPs under fluorescence microscope. The in vitro cytotoxicity of will be examined on breast cancer cell lines and control cell lines: Se cocktail; Naked NPs; Se cocktail loaded naked NPs; NPs with targeting antibody; Se cocktail loaded NPs with targeting antibody.
Speciation of selenium before (after loading targeted nanosystems, drug release, stability) and after cell uptake.
WP3 Pre-clinical study design and Evaluation
Based on the requirements for the evaluation, which have to be determined first, an evaluation plan will be elaborated. Based on the needs for the preclinical stage, methods for evaluation will be selected. The data created in WP 1 and WP 2, including Se-compounds encapsulation, antibody production and coupling, will be evaluated in WP 3. Results of evaluation will be presented.
The therapeutic effect of the nanosystem developed in WP 1&2 will be evaluated according to T3.1. plan, in a syngeneic mouse breast cancer model and a human breast cancer model developed in nude mice.
Nanoparticle biodistribution profile will be studied in normal mice breast cancer bearing mice. It will be monitored by in vivo NIR fluorescence imaging at 1 h, 4 h, 24 h and 48 h after injection. Mice will be killed at each time point and we will proceed to ex vivo NIR fluorescence imaging of their organs.
For the toxicity study in mice, repeated daily dosing over 28 days with a 2-week recovery period will be required. The study includes formulation analysis, daily observation of clinical signs, measurements of body weight and food consumption; ophthalmoscopy, hematology, blood chemistry and urinalysis, and histopathology. The data created in the preclinical stage will be evaluated in WP 3. Results of evaluation like statistical analysis will be presented.
WP4 GMP production & quality control
To understand procedure of operations used, critical and non-critical parameters of each operation, production environment, equipment and excipient availability which should be taken into account during the early phases of development of formulation, so that successful scale up can be carried out. To enhance drug quality as developed by R&D in final formulation as well as to assure quality for predetermined period of time. To provide information on the selection of materials and design of procedures on the basis of innovator product characteristics through tests and compatibility studies. To identify specification and quality.
Set up production protocols according to GMP guidelines. Procurement of GMP quality raw materials (PLGA, PVA), identification of specification and quality. Establish production protocol of NEOSETAC NPs according to GMP guidelines including formulation. Definition of the required quality controls and design of stability testing SOPs conforming GMP and medical requirements.
Establish large-scale production protocol of Se-NPs according to GMP guidelines. Quality control on large-scale produced NPs (physical properties, storage condition and shelf life stability). Validation and qualification of premises, supporting utilities, and equipment (VEC) Establish criteria and frequency for validation and qualification. First batch of Se-nanosystems produced, characterization, quality control and stability studies
WP5 – Communication, dissemination and networking
Communication, dissemination and networking activities will serve NEOSETAC project covering its different steps, to secure the enrollment of key stakeholders, the enlargement of NEOSETAC network and a wider dissemination of key achievements. In this respect, work-package 5 aims to carry out a set of actions aiming at increasing the visibility of NEOSETAC project, partners, and activities as well as spreading and exploiting the project results in order to make it a successful and sustainable project.
WP6 – Project Management
This work-package allows an efficient management at administrative, contractual, legal, scientific and technical levels towards delivering high-quality results within the project scope, schedule, and cost, whilst taking into account stakeholders satisfaction and potential project risks. Internal monitoring and self-evaluation mechanisms, based on a well-defined Project Quality Handbook and risk management of all foreseen activities have been established. An External Advisory Board will oversee the project progress and assist as a controlling mechanism
This project is co-funded by the European Union’s Horizon 2020 Research and Innovation Programme under Reference N. H2020-MSCA-RISE-2017 GA778325
Prof. Manuel Valiente
Universitat Autònoma de Barcelona,
Science Faculty. Building
(+34) 93 581 49 38