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Report

Teaser, summary, work performed and final results

Periodic Reporting for period 2 - FOLSMART (Folate-Target Nanodevices To Activated Macrophages For Rheumatoid Arthritis)

Teaser

Summary

In NANOFOL (FP7 project running from 2009-2013) we have developed novel folate receptor-targeting liposomes by incorporating a bifunctional SP-DS3 peptide consisting of the neck domain of surfactant proteins, which serves both as a linker and an anchor of the folate to the liposomal membrane. Complete prophylactic efficacy against arthritis was observed in mice treated with these liposomes encapsulating methotrexate (MTX). The currently used drug therapy for rheumatoid arthritis (RA) with MTX has shown lower clinical scores in mice models compared to the novel folate-based liposomes (FBL) (Figure 1).
The demonstration of reproducibility and stability of larger scale batches (GMP) with reference to lab batches in terms of both chemical composition and function is imperative to fulfil one of the main goals of Folsmart: to bring FBL to Phase I clinical trials as a treatment option for RA. WP2 intends to develop safe methods for large-scale production of the liposomal formulation with FA-peptide linker and encapsulated MTX under GMP. Also, alternative pH-responsive liposomes are being used to tune the MTX release at the inflamed target site.
The in vivo efficacy of the proposed FBL will be tested in animals in WP3, to prepare for the First-in Humans clinical trials. First, we will use a reference mouse model of RA, the Collagen-Induced Arthritic (CIA) mice, which reproduces some features of the human pathology and allows the evaluation of the efficacy of the treatment. The non-clinical toxicology and safety package will be performed in rat and dog under GLP. Data from these studies and from the GMP manufacturing will compose the dossier for the request to INFARMED for authorisation to implement the Phase I clinical trials in healthy subjects (WP4).
The objective of WP5 is to present the business plan and pharmacoeconomic evaluation for this new therapy. Therefore, FBL therapy will be benchmarked against MTX (free drug injectable solution, usual dose 15 mg/week). The competitiveness of Folsmartâ€™s approach is envisioned as FBL being an alternative therapy to biological DMARDS, delaying or avoiding these options when MTX is not/no longer recommended. The developed formulation will be highly engineered to become a therapy with optimal performance, hence it shall lead to a better clinical outcome, lower toxicity levels and fewer side effects.
Two WPs dedicated to project management (WP1), communication, dissemination and exploitation (WP6) ensure the objectives of the Project are timely fulfilled and its outcomes are efficiently disseminated throughout the scientific community and general public. IPR over Folsmartâ€™s innovation will be carefully handled.

Work performed

The method for the production of Folsmartâ€™s FBL was modified by Tecminho in order to increase the amount of encapsulated MTX. After implementation at lab-scale, a pilot scale unit was assembled and the manufacturing method was optimised to become reliable, reproducible, scaled-up and certified under GMP. Tech transfer is being handled from Tecminho to Bluepharma, responsible for the GMP production, with the inherent analysis and validation battery of tests. Several alternative approaches of MTX encapsulation were assayed by Tecminho and BOKU. The mechanisms of drug release are being explored by BOKU, using pH-responsive peptides anchored to the liposomes.
The efficacy of FBL was confirmed in vivo by INSERM in CIA mice models in comparison with the free MTX injectable solution. Healthy mice were challenged with the same FBLs by Synovo to evaluate the biodistribution profile and toxicity. Liposomal MTX has a much longer half-life in circulation than the free drug, as expected. The distribution pattern seems dominated by filtering organs like the kidney. The loss of appetite and decrease on the body weight were early signs of toxicity. Further evaluation has indicated that a possible target site is the gut epithelium and the bone marrow, both highly proliferative tissues. The use of subcutaneous application combined with lower doses should moderate this issue; INSERM is evaluating this strategy.
The MoA in vivo is being assessed by INSERM using bioimaging tools and flow cytometry of immune cells transferred from healthy mice to RA mice. The immunomodulatory properties of the FBLs are being comprehensively tested by MUW and IBMC using primary human monocytes, FR+ macrophages, and T cells. In parallel, MUW and Tecminho worked on alternative functionalization methods with anti-FR recombinant monoclonal antibody fragments. However, these FR-Fab fragments seem not resistant to the conditions being now used in the FBL production, as the specificity of FR-Fab-tagged FBL for FR+ macrophages was not superior to the one observed for FR- macrophages.
At the beginning of 2018, Dose Range Finding studies were carried out by Aptuit in rat and dog which showed the limiting toxicity and established the maximum tolerated dose. Immunogenicity was also assessed as the potential for anti-drug antibody formation (ADA). Subsequently, pivotal GLP studies to assess toxicity and toxicokinetics following repeated IV administration in rat and dog have been initiated; the in-life phase will be completed by mid-March. In addition, a biodistribution study to assess the pharmacokinetics and the tissue distribution of the total MTX and its major metabolite in rats has been performed.
Blueclinical prepared and submitted the documentation for the request of Regulatory and Scientific Advice to INFARMED regarding the Folsmartâ€™s FBL. Additional evaluations/studies were implemented in order to address some points that should be adequately covered.
The implementation and conduction of the First-in-Human clinical trial are foreseen to start in the second semester of 2019. Blueclinical is working on the development of the documents needed for submission.
A thorough study of the market and pharmacoeconomic evaluation has started at the end of 2018, with the main competitors being free MTX injectable solution and biological DMARDS.

Final results

RA affects over 2.9 million people in Europe, nearly 50 million people worldwide. The goal of RA treatment is to alleviate the current symptoms and to prevent the destruction of the joints and the possible handicap. Folsmartâ€™s new therapeutics will allow treatment efficacy with reduced doses, fewer side effects and possibly better patient compliance. Furthermore, the use of the new technology is expected to cut the costs associated with biological therapies at least by 50%, by delaying or avoiding its administration.
Folsmart will provide favourable conditions for industrial innovation, ensuring that R&D is translated into effective and safe wealth-generating products. The manufacturing process is a key step in the establishment of the new technology. By modifying several steps and conditions on the former production method, FBLs are now encapsulating over 10 times more the amount of MTX previously incorporated. This inherently leads to a great waste reduction, which in the case of MTX, being a potent drug, is even more significant.
The nonclinical safety studies will define the pharmacological and toxicological thresholds prior to the initiation of human studies. If no concerns are raised during the nonclinical tests, the First-in-Human clinical trial in humans testing the developed novel FBL in healthy subjects is programmed to the second semester of 2019.