Periodic Reporting for period 1 - EDROMA (Novel nanomedicine for the treatment of glioma) Reporting period: 2018-03-01 to 2018-08-31 Summary of the context and overall objectives of the project Problem/issue addressed by the projectLisapharma, an Italian pharmaceutical SME, aims to bring on the market Edroma, a new product based on a nanotechnology platform to treat malignant high-grade gliomas (HGG), in particular glioblastoma (GBM). GBM is the most aggressive type of brain cancer for which there are no satisfactory curative therapies. The Edroma medicinal product is a dispersion for injection made of nanoparticles loading the active ingredient zoledronic acid, obtained by a self-assembling procedure patented by University of Naples (ITA) and exclusively licensed to Lisapharma. Compared to the current brain-cancer therapy solutions, Edroma demonstrated in animals significant improvements characterized by higher efficacy and lower toxicity.Importance for societyDespite recent advances in treatment, the prognosis for patients with HGG is dismal; so far the most effective treatment, i.e. radiotherapy, brings an increase in median survival from 3-4 months to around 9-10.Only modest advancements in the treatment of GBM have occurred in the past 25 years, due to two main problems: 1 ) No specific laboratory studies are helpful in diagnosing GBM. Imaging studies of the brain are essential but, in most cases, complete staging is neither practical nor possible. These tumors do not have clearly defined margins; they tend to invade locally and spread along white matter pathways, creating the appearance of multiple GBMs or multicenter gliomas on imaging studied. This makes early diagnosis so infrequent that treatment is usually unable to provide a complete cure. 2 ) Medical therapy primarily involves glucocorticosteroids, which often produce a marked improvement in neurological symptoms by themselves. Chemotherapy has been used for primary therapy as either single agent or multi-agent regimens. Results have generally been conflicting and significant systemic toxicity is possible. These methods have not so far demonstrated satisfactory results, prolonging the survival only of few months, if compared to surgery (not possible in most cases) and radiotherapy alone.Overall objectivesThe general objective of this Phase I project was the development of a detailed Business Plan for attracting investments for the Edroma nanomedicine product development, registration, industrialization and introduction on the market, which includes activities related to detailed measurable specific objectives listed below:OBJ 1: Product validation and industrialization planning, including: i) planning of preclinical and clinical studies based on the Protocol Assistance document received by the European Medicines Agency (EMA) on approval of Edroma’s orphan designation (EU/3/16/1735), in which the detailed preclinical and clinical trials to execute are described; ii) industrialization requirements assessment and planning, for all the activities and resources required to transfer the production of the new product to the selected manufacturing plants operating in Good Manufacturing Practice (GMP) conditions.OBJ 2: Commercialization planning and partnerships definition, including: i) market analysis to assess the existing and potential market (size and growth) of HGG cases that might be treated through the new brain-targeted pharmacological nanosystem medicine; detailed commercialization and dissemination planning, taking into account the necessity to distribute the product at international level, given the relatively low number of HGG cases, and to reach a highly specialized audience of customers, i.e. clinicians working in cancer care hospital departments, as well as distribution partners; potential commercialization agreement definition, in particular signing license & supply agreements that appears as the more realistic strategy to develop the business.OBJ 3: Definition of the legal (contractual) and ethical framework, including: i) assessment of Ethical Issues and regulations affecting product distribution in the different national markets, e.g. in terms of evaluation of the therapeutic options by examining the “good and harm” factors, the respect of the equality rights of the patients and the service to the community; ii) definition of the legal architecture to manage contractual relationships with partners and institutions, taking into account of the related ethical issues, and of further IPR protection strategy for the product after the orphan medicine exclusivity period.OBJ 4: Financial plan preparation, including: i) quantification of the revenues and costs based on the detailed market analysis and on the cost of clinical validation, industrialization, commercialization, production and distribution activities; ii) financing requirements and Return On Investment evaluation by forecasting the cash flow and projecting financial performance indicators on a 5 years period, and by identifying the planned sources of funding. Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far The work performed in this project was the preparation of a business plan for Edroma, Lisapharma’s medicine for glioblastoma, based on detailed feasibility studies and investment plan for the nanomedicine product development, registration, industrialization and introduction on the market. The present feasibility study, in particular:✓ Presents findings of the market and landscape analysis,✓ Articulates existing regulatory pathways for Edroma,✓ Proposes a development and commercialization plan for Edroma✓ Reports the commercial value assessment and financial business plan of Edroma.The feasibility study was created through the following methodology:1) Analysis based on secondary research, Lisapharma and Alira Health (AH) expertise.2) Secondary research using various sources, including public and non-public available information.3) Specific competences (Technical; regulatory; industrial and commercial) acquired attending international events on Orphan Medicines such as World Orphan Drug Congress. This is the marketplace for Orpharn Drug professionals looking at the complete value chain of orpharn drug development, from clinical development and R&D to market access. The geographical scope of the analysis was EU and US markets. Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far) The medicinal product Edroma is a dispersion for injection made of nanoparticles encapsulating the active ingredient zoledronic acid (ZOL). This nanosystem has a core-shell structure, consisting of calcium phosphate/zoledronic acid nucleus coated by two bilayers of transferrin-targeted PEGylated lipids. In detail, the core of the nanosystem is made by calcium phosphate (CaP) binding zoledronic and the shell by two lamellae of positive lipids bilayers, in part PEGylated. The external lamella is non-continuously coated, i.e. “decorated” , by the human transferrin as ligand, electrostatically bound to the lamella. This nanosystem product, or nanovector, coded TF-PEG-CaP/ZOL-NPs, is prepared by a self-assembling procedure, shown in Figure 1. The resulting colloidal dispersion is formed by agitation giving rise to NPs with a mean size of about 150 nm. The formulation has been developed to obtain a nanovector with a high ZOL loading. Indeed, when using “conventional” nanovectors, e.g. stealth liposomes, the drug encapsulation efficiency is below the 5-6% of the ZOL initially used in the preparation, while Edroma nanoparticles are characterized by a ZOL encapsulation efficiency of 100% . Indeed, the inventors demonstrated that the use of nanotechnology-based formulations overcomes the limitations due to the bone preferred zoledronic distribution, thus conferring to this drug its powerful anti-cancer activity. Edroma introduces significant benefits, as confirmed in the EMA orphan designation report:- Effectiveness. The efficacy of the product proposed in this application has been demonstrated in different animal models, among them two different experimental models of HGG. Mice were treated i.v. with blank nanoparticles with transferring (NPs-Tf), NPs-ZOL and NPs- ZOL-Tf (Edroma product) at 20 µg/mouse for three time a week, for 3 consecutive weeks. It is evident that NPs-ZOL-Tf exhibited the highest antitumor efficacy. In fact, as summarized in Figure 2, this treatment produced, at nadir of the effect, a significant tumor weight inhibition of 41%, while the non-functionalized NPs-ZOL reduced of 31% the growth of tumors and free ZOL resulted in a not particularly marked tumor growth inhibition (TWI 20%). The good therapeutic efficacy of NPs-ZOL-Tf is also demonstrated by the significant delay of tumor growth (10 days) and by the increase of life survival of mice (23%). Interestingly, this treatment produced a complete tumor response in 1 out of six mice treated, while NPs-ZOL produce a stabilization of disease in 1 out of six mice treated. It is worthy of note that the complete disappearance of the tumor persisted for more than six months after the end of treatment, as confirmed by imaging analysis. This increase in survival of mice treated with NPs-ZOL-Tf is impressive considering that the median survival time of untreated experimental group was of about 1 month and also in comparison with the life expectancy of healthy mice of the same strain used in these experiments, that is of about 2 years.- Low toxicity. Besides increasing the effectiveness of chemotherapy, Edroma ZOL-loaded nanoparticles allow avoiding the toxic side effects associated to other treatment methods currently applied on HGG patients. In the above-mentioned experiments, all treatments were well tolerated by the animals, as there were no toxic deaths or weight loss in any of them. The instability of therapeutic nanoparticles in the blood is considered to be one of the serious limitations in their clinical use. In particular, carriers based on cationic lipids often present hemolytic activity at certain concentrations. Edroma nanoparticles showed almost no hemolytic activity (less than 2%), regarded to a non-toxic effect level according to ASTM F 756–08. A further important goal in the preclinical drug development was to assess whether Edroma formulation would be clinically toxic to bone marrow. Bone marrow is critically sensitive to many cytotoxic agents and myelotoxicity represents one of the limitations to the use of full doses of antineoplastic drugs, including ZOL. Pre-clinical tests of Edroma on mice demonstrated tumor reduction, increase of survival and even, in one case out of 6, complete cure. Lisapharma has applied and obtained the orphan designation of the medicinal product from both EMA and FDA. This represents a unique business opportunity, as Lisapharma will be the only authorized provider of the registered product for 10 year (7 in USA).