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WE ARE INNOVATIVE

“RESEARCH – CREATE – INNOVATE” INITIATIVE

PROJECT TITLE

Innovative nanocarriers for the targeted and prolonged treatment of cancer and demyalinating diseases of the central nervous system

ACRONYM

NANO-MYTHiC

PROJECT CODE: 

Τ2ΕΔΚ-00501

Project Description

Nowadays, more than 1.5 billion people worldwide suffer from neurodegenerative disorders such as multiple sclerosis, Parkinson's, Alzheimer's, and others. Neurodegenerative diseases are the fourth leading cause of death and 10 million new cases per year are reported. The development of therapeutic agents that could enhance re-myelination or prevent degeneration is a major challenge for neuroscientists. Recently, antibodies to two myelin-derived neuronal regeneration inhibitors have been developed, particularly Nogo-A (Ozanezumab) and LINGO-1 (Opicinumab). The inhibition of the above proteins has beneficial effects on demyelinating animal models and specific antibodies are used in clinical studies aimed at re-myelinization and neuroprotection. Moreover, cancer is the second cause of death after cardiovascular diseases, and prostate cancer (PCa) is the most prevalent cancer in men with 750,000 new cases and 140,000 deaths every year in the developed world. Many genetic alterations mainly to the Akt / PI3K and RAS / ERK signaling pathways are associated with the disease, and the most common are the ETS gene chromosomal rearrangements. Treatments with anti-androgens are effective in early stages but in later stages or on metastatic tumors. The lack of animal models that closely approximate the progression of the disease and the lack of new therapeutic agents other than androgens is one of the main problems in dealing with advanced stages of the disease and high mortality. Hence, both fields are of intense activity in the pharmaceutical industry.

In this context, effective and targeted administration with suitable biocompatible polymeric carriers is a feasible goal to produce new and innovative pharmaceutical formulations. The polymers are widely used as active pharmaceutical drug delivery vehicles and almost all the novel formulations currently produced contain biocompatible polymers. This project aims to develop new controlled release drugs for some of the most serious diseases with enormous socio-economic significance and burden, such as neurodegenerative diseases and cancer. Many therapeutic approaches are in preclinical or clinical trials for both diseases and include new drugs, drug reformulation and pharmacokinetic optimization of existing ones. Despite the progress made in recent years, effective tackling of these diseases remains a challenge, and even small improvements have significant social and economic impact.


 

Object and Objectives of the Project

The aim of this project is to test new compounds such as anti-LINGO and anti-NOGO antibodies and hydrophobic inhibitors of MEK/MAPK with proven anti-cancer and/or anti-degenerative activity, and to prepare novel pharmaceutical formulations for their targeted and controlled administration, in order to offer new directions and tools in the field of neurodegenerative disorders and cancer treatment.

Taking advantage of the available experimental models, technological advances and industrial processes, the therapeutic effect of new compounds with anti-cancer and/or anti-demyelinating activity will be investigated, in order to enhance and further specialize the activity of existing pharmaceutical compounds, as well as to optimize their production process. Specifically, the individual objectives are:

  1. Development, testing and production of innovative polymeric carriers, micro-/nanoparticles and patches, which will aim to the targeted and controlled release of new active therapeutic agents such as the anti-LINGO and anti-NOGO antibodies for the treatment of myelinopathies, and MEK/MAPK inhibitors, such as U0126 and GSK1120212 (Tramenitib) to treat prostate cancer.
  2. Comparative evaluation of the effect of the designed polymeric carriers and the active compounds in free-form on a treatment-responsive ERF-induced craniosynthesis model.
  3. Use of an ERF gene-based animal model developed by IMBB-FORTH in pharmacological studies on adult prostate cancer.
  4. Study of genetic and toxic models of demyelination and remyelination after the administration of new therapeutic substances, which will open an important field for monitoring innovative therapeutic approaches.
  5. Investigation of the molecular mechanisms involved in myelinopathies and prostate cancer.
  6. Development of a new method for drug identification and selection that exploits the molecular modification of the studied genes.
  7. Pilot production of the final formulations to be used commercially by PHARMATHEN SA and the project partners.

From the above objectives, it is clear that the subject of this project is in the priority area for Health and Medicines, and especially in the thematic area concerning the development of generic value-added products, with a particular priority the strengthening of innovation in the production process of the medicinal product. In addition, the utilization of the results of the proposed project will directly contribute to the development of new pharmaceutical products of high added value and exportability, which is also an overall goal of the program.


 

Project Implementation Methodology

NANO-MYTHiC is a 30-month interdisciplinary project with three (3) involved partners: the Institute of Molecular Biology and Biotechnology of the Institute of Technology and Research (IMBB-FORTH), the Laboratory of Chemistry and Technology of Polymers and Dyes of the Department of Chemistry of the AUTH (AUTH) and PHARMATHEN International SA, each one bringing forward their experience in biotechnology, polymer science and pharmaceutical technology, respectively.

For the implementation of the project, the three partners will combine their expertise in order to achieve maximum efficiency within the proposed timeframe, aiming for pre-industrial prototyping and production of anti-cancer and anti-demyelinating formulations. IMBB laboratories use the available cell and animal models to evaluate the initial pharmaceutical effect of the bulk compounds, while at the same time AUTH laboratory in collaboration with PHARMATHEN will optimize their encapsulation process. The drug-encapsulated formulations are then tested on the available biological systems both for their effectiveness and for the possibility of industrial scale-up. Finally, their standardization and commercial registration is being prepared.

A series of compounds that have been priorly tested at an experimental, preclinical and even clinical level, showing an anti-cancer and/or anti-demyelinating activity, will be further studied on demyelinating animal models, as well as in corresponding cell systems. The proposed strategy for demyelinating diseases is to more effectively deliver antibodies (anti-LINGO and anti-NOGO) that neutralize myelin inhibitors, whereas for prostate cancer we will use inhibitors of both the MEK/MAPK pathway, which have been shown to enhance the effect of ERF. For the administration of these compounds, two (2) polymeric micro-/nanoparticle delivery systems of fast (up to 1 week) and slow-type (up to 2 months) release for intravenous use will be developed. The proposed systems will be prepared from PEAd-co-PLGA copolymers, newly synthesized in AUTH laboratory. Alternatively, a third delivery system that will involve the preparation of a sustained-release patch for the transdermal administration of MEK/MAPK inhibitors will be designed. To evaluate the impact of encapsulation in terms of improved bioavailability and cytotoxicity, but at the same time to reduce the use of animal models to the absolute necessary extent and to balance the cost of materials for molecular analysis, the encapsulated compounds will be initially tested in cell models. Those that meet the desired kinetic characteristics will also be used in laboratory animals.

The final stage of the project will include the encapsulation of the proven active compounds in a pilot stage at PHARMATHEN International SA facilities. There will also be an optimization of the parameters according to the standards applied by the company and a study of the pharmacodynamic characteristics.

 

Description of work units and deliverables

1. Synthesis and characterization of biocompatible polymers
Preparation of poly(ethylene adipate)-co-poly(lactic-co-glycolic acid) (PEAd-co-PLGA) copolymers containing 10-90 wt% PEAd. Synthesis of modified chitosan derivatives. Characterization of polymeric materials.

Deliverables
1.1 Polymer Synthesis Protocols, 1.2 Polymer Characterization Report

2. Preparation and study of nanoparticles
Preparation of new formulations of anti-LINGO, anti-NOGO, U0126 and GSK1120212 compounds:
-    PEAd-co-PLGA nanoparticles with TPGS as emulsifier for brain targeting
-    Polyelectrolyte capsules with Layer-by-Layer (LbL) technique for micro/nanoencapsulation
-    Patches with PEAd- co-PLGA and modified chitosan for transdermal delivery 
-    Physicochemical characterization of formulations 
-    Study of the in vitro release rate of encapsulated drugs

Deliverables
2.1 Micro/nanoparticle and patch preparation protocols, 2.2 Micro/nanoparticle and patch characterization results, 2.3 Drug release diagrams, 2.4 Publication of results at scientific conferences

3. Release and localization tests of the substances under study in vitro and in vivo
Determination of the optimal conditions for administration of the studied compounds ex vivo and in vivo: Endocytosis of encapsulated drugs, Kinetic and biological stability of MEK and XPO inhibitors, Pharmacokinetic study of the encapsulated compounds

Deliverables
3.1 Administration protocols of the encapsulated substances in cells and normal muscles

4. In vivo administration in controls and models of demyelination (genetic and toxic)
Monitoring the in vivo effect after the administration of the studied encapsulated compounds on animal models of demyelination/remyelination: Genetic models & Toxic model

Deliverables
4.1 Report effect of encapsulated compounds on ERF loxP/-; Nestin-Cre +/- genetic model and on the model of toxic demyelination by administration of the toxin Cuprizone, 4.2 Publication of results in an international peer-reviewed journal

5. Optimization of animal models for prostate cancer
Determining the minimal genetic changes that best simulate the disease progression in human prostate cancer
Deliverables
5.1 Animal models of prostate cancer

6. Effects of compounds that enhance the action of ERF in prostate cancer
Determination of the minimum dose and administration protocol of compounds for prostate treatment: Animals with craniosynostosis & Animals with prostate cancer

Deliverables
6.1 Compounds affecting prostate cancer, 6.2 Publication of results in an international peer-reviewed journal

7. Identification of Molecular Targets
Identification of gene expression changes in target cells.

Deliverables
7.1 Report on the effect of encapsulated compounds on the molecular profile of oligodendrocytes, epithelial and mesenchymal cells after the effect of nanoparticles. Potential list of target genes for further therapeutic approaches on demyelinating pathologies and prostate cancer

8. Preparation of a pharmaceutical preparation on a pilot scale
Pilot preparation of PEAd-co-PLGA micro/nanosphere formulations

Deliverables
8.1 Protocols for pilot preparation of formulations, 8.2 Report of drug encapsulation results, microsphere characterization and release rate

Expected results

NANO-MYTHiC project is expected to vastly contribute towards the direction of advancing the current knowledge and methodologies for treating demyelinating diseases and cancer by proposing innovative delivery systems. Novel polymeric drug-carriers will be designed by combining the recent technological advances in nanotechnology for localized, long-term and controlled drug release. The proposed research and innovation of the NANO-MYTHiC project covers all the goals of the "RESEARCH - CREATE - INNOVATE" program, while the expected benefits for PHARMATEN SA and the participating academic institutions are multiple:

•    Development and production of new polymeric carriers, which will aim at the targeted and controlled release of new active therapeutic agents.
•    Development of new expertise and fostering the relationship between research and production to transfer this technique to production level so that commercially available products can be produced.
•    Development of synergies and partnerships in fields of common interest.
•    Increasing productivity and replacing inefficient formulations.
•    Improving the position and competitiveness of PHARMATEN SA in the international market with the production of innovative products.
•    Reinforcement of the company's strategic planning at national and European level for expansion, modernization and promotion of existing infrastructure and provided innovative services.

The scientific results of the proposed NANO-MYTHiC research proposal for the development of innovative drugs for the effective treatment of myelopathies beyond synergy are:

•    Multidisciplinary approach, with researchers covering a wide range of scientific fields such as chemistry, biology, pharmaceutical technology, materials technology, production of innovative formulations, etc.
•    Improvement of patients' quality of life due to the advantages of the proposed formulations.
•    Increase patient compliance.
•    Maintaining the optimal concentration of drugs with targeted action and increased duration of the therapeutic effect and increasing patient compliance.
•    Enhanced safety of drug administration of drugs (e.g., avoiding overdose).
•    Reduced costs for both care provider and patient.