The invention relates to pharmaceutical compositions comprising lipid- bacteriophage conjugates, wherein lipid, as the glycolipid alpha-Galactosylceramide is immunologically active and the bacteriophage is engineered to bind to target cells and carry antigenic molecules capable of stimulating an immune response. Such bacteriophage is used to stimulate an immune response against cancer and against infectious agents.
The economic relevance of "cancer immunotherapy" strategies is high.
This is an expanding market whose limits are difficult to predict. Vaccines currently represent the largest segment (≈20%) of the global biopharmaceuticals market (≈ $ 163 billion) and the global cancer vaccine market was valued to be around $4,188 milions in 2019
With this new technology we aim to contribute to the formulation of new immunotherapeutic strategies against cancer and infectious agents.
To obtain better results in the fight against cancer, different immunotherapies should be used in combination. Alongside the need to improve the effectiveness of the available strategies, there is the urgence to propose new tools that can be quickly developed and at low costs. We propose a technology for the formulation of natural nanoparticles based on filamentous bacteriophages delivering protein
antigens together with immunologically active lipids and that can be delivered to specific target cells.
Our proposal aims to implement the existing combined therapies to attack cancer cells on several fronts and to adopt innovative and more effective therapeutic strategies capable of counteracting the various escape mechanisms used by cancer cells.
Current Technology Limitations
The main interventions in the field of immunotherapy (CAR, adoptive T cell therapy; monoclonal antibodies) have relatively high production and preparation costs. New technological implementations should reduce costs as well as expand therapeutic efficiency of the proposed therapy.
The technology proposed by us provides low production costs and higher safety profile. Furthermore, it is known that to fight cancer, different immunotherapeutic strategies should be used in combination, therefore there is both the need to improve the effectiveness of the yet available strategies and to propose new strategies that can be rapidly implemented.
At the moment, there are no immunotherapy products derived from engineered bacteriophages on the market. However, given the enormous and emerging need to find new technological strategies for cancer immunotherapy, the proposed formulation based on the use of recombinant phages conjugated to immunologically active lipids may assume enormous scientific and industrial importance. This phage formulation combines pharmacological efficacy with safety administration in humans and with the industrial production at advantageous costs, and it could be rapidly used as a therapeutic agent to induce immune responses against cancer in patients. We are currently focused on a preclinical study based on a melanoma model using phages co-expressing Tumor Associated Antigens together with the antitumor lipid a-GalCer and able to be delivered specifically to tumor cells.
Our Technology and solutions
We propose the formulation of natural nanoparticles based on filamentous bacteriophage fd, a non-lytic and non-pathogenic virus, for the delivery of immunologically active lipids, such as (but not exclusively) the glycolipid alpha-Galactosylceramide (α-GalCer). These lipids bind the phage coat proteins through hydrophobic interactions. Using the phage display technology, the bacteriophages can be also engineered for the delivery of short tumor antigens in high copy number. Indeed, we have demonstrated that therapeutic vaccination with these bacteriophages is able to inhibit the tumor growth in animal models. Furthermore, the proposed technology allows to construct bacteriophages capable of being specifically targeted to different dendritic cell subpopulations, to increase the recognition and immunogenicity of the delivered antigens.
Bacteriophages infect and multiply exclusively in their hosts (bacteria) and are widespread in the environment. For these reasons, their administration in humans can be considered safe. Furthermore, their production is relatively simple and economically advantageous. In previous studies we have demonstrated both the ability of phage nanoparticles to stimulate innate immunity and to induce a strong adaptive response to the delivered epitopes. More recently, we have shown that the co-expression of tumor-associated antigens and a-GalCer on phage nanoparticles induces a strong iNKT response and the production of Cytotoxic lymphocytes with specific antitumor activity. This technology thus represents an innovative approach to enhance anti-tumor responses, overcoming the current limits associated with the use of α-GalCer in therapy.
For the industrial development of the technology, a drafting of a Standard Operating Procedures for the preparation of phages on a laboratory scale will be settled. The production process of bacteriophages will therefore be translated on an industrial scale by fine-tuning the production process using special fermenters, and optimizing the growth conditions such as temperature, nutrients, and oxygen, and the purification conditions. Furthermore, the project will provide the development of suitable chemical and biotechnological strategies for pre-GMP manufacturing of the bacteriophage-based vaccine that has obtained the highest efficacy in preclinical studies, according to the parameters described in the guidelines of the Europen Medical Agency (EMA), for subsequent phase I studies in humans.
The patented technology has already been validated in the laboratory. We have collected evidences that the co-delivery of the α-GalCer glycolipid and a tumor associated antigen on the surface of the filamentous phages allows the enhancement of the adaptive anti-tumor immune response and results protective in animal models of therapeutic vaccination, confirming a TRL3 / 4 level of technological development. Geographical coverage of the patent was extended through a European and US patent application.