Periodic Reporting for period 1 - HydroTheC (Biodegradable and Eco-friendly Smart Hydrogels for the Sustained Intraperitoneal Release of Chemotherapeutics as a Novel Local Therapy Approach for Advanced Ovarian Cancer)
Reporting period: 2024-01-01 to 2025-12-31
The general goal of the HydroTheC project is the development of a new non-toxic, natural, biocompatible, biodegradable, thermo-responsive, and injectable multidrug-loaded HG as an in situ depot system for the sustained intraperitoneal release of chemotherapeutics for ovarian cancer (OC) treatment. This new therapeutic tool is crucial to control the abdominal spread of reminiscent OC cells after the surgery, avoiding OC metastization, but can also be applied in the case of unresectable tumors, or as a coadjutant therapy for tumor reduction before its surgical removal. The designed formulation will ideally contribute to increasing the drugs' therapeutic window and avoiding the appearance of resistance to the therapy. Moreover, this local therapy approach would reduce the systemic side-effects and the number of needed administrations, which tremendously increases patients' welfare and contributes also to a reduction in hospital costs. Another general objective of HydroTheC is the creation of a “green” formulation, in terms of its composition (avoid petroleum derivatives) and preparation method, but also in terms of the environmental impact of its development, by using in silico prediction tools in the formulations optimization phase. Also, by using simple and low-cost production methods, as well as polymers and drugs already approved for human use, another goal of HydroTheC is to achieve a formulation that is easy to scale-up and translatable to the clinical practice.
To achieve this, the specific objectives of HydroTheC are:
1. Selection of the formulation components
2. Use of in silico bioinformatic tools
3. Physic-chemical characterization of the formulation
4. Assess the biocompatibility and efficacy of the formulations in co-cultured and 3D in vitro models
5. Quantitative and qualitative determination of the biodistribution, pharmacokinetic, and therapeutic impact in terms of OC regression in in vivo models
6. Study the scale-up and stability of the formulation (e.g. sterilization, lyophilization, storage, etc.), as well as its possible translatability
7. Dissemination of the project results with the scientific community, citizenship, and potential stakeholders
To achieve this, the specific objectives of HydroTheC are:
1. Selection of the formulation components
2. Use of in silico bioinformatic tools
3. Physic-chemical characterization of the formulation
4. Assess the biocompatibility and efficacy of the formulations in co-cultured and 3D in vitro models
5. Quantitative and qualitative determination of the biodistribution, pharmacokinetic, and therapeutic impact in terms of OC regression in in vivo models
6. Study the scale-up and stability of the formulation (e.g. sterilization, lyophilization, storage, etc.), as well as its possible translatability
7. Dissemination of the project results with the scientific community, citizenship, and potential stakeholders
Work Performed and main achievements at the end of the 1st year of the project:
1. Selection of the formulation components: For the accomplishment of this objective, the polymers used for HG formulation were chosen from a library of possible compounds originated from waste materials or produced by natural sources in order to fulfill the requirement of creating an eco-friendly formulation. Also, for micelles production was used Soluplus®, a polymer that is approved for human use.
2. Use of in silico bioinformatic tools: Bioinformatic tools were used during the first year in several tasks of the project: i) To predict the behavior of the polymers at different biological pHs, ii) to predict the structure of the polymeric micelles and their interaction with the hydrophobic drug, iii) to find, among ovarian cancer patients databases which are the most relevant biomarkers to perform a targeted therapy, and iv) to predict the structure of a peptide that can be attached to the nanoparticles surface and promote an active targeting against the cells of interest.
3. Physic-chemical characterization of the formulation: The designed formulation was characterized in terms of chemical interactions, morphology, viscosity, thermo-responsiveness, and rheologic behavior.
4. Assess the biocompatibility and efficacy of the formulations in co-cultured and 3D in vitro models: Despite this part was not planned for the first year of grant, the biocompatibility of the designed formulation was tested in 2D healthy fibroblasts models, and preliminary results of in vitro efficacy were also performed in a 2D ovarian cancer cell model. Moreover, the optimization of the cell culture conditions for the growth of ovarian cancer 3D spheroids was also initiated.
6. Study the scale-up and stability of the formulation (e.g. sterilization, lyophilization, storage, etc.), as well as its possible translatability: The tasks associated with this objective were scheduled for the third year of the grant; however, it was tested the possibility to lyophilize the HG formulation.
7. Dissemination of the project results with the scientific community, citizenship, and potential stakeholders. The tasks associated with this objective were scheduled for the entire period of the grant. During the first year, some dissemination actions of the work were performed in scientific conferences and also in actions for citizenship.
1. Selection of the formulation components: For the accomplishment of this objective, the polymers used for HG formulation were chosen from a library of possible compounds originated from waste materials or produced by natural sources in order to fulfill the requirement of creating an eco-friendly formulation. Also, for micelles production was used Soluplus®, a polymer that is approved for human use.
2. Use of in silico bioinformatic tools: Bioinformatic tools were used during the first year in several tasks of the project: i) To predict the behavior of the polymers at different biological pHs, ii) to predict the structure of the polymeric micelles and their interaction with the hydrophobic drug, iii) to find, among ovarian cancer patients databases which are the most relevant biomarkers to perform a targeted therapy, and iv) to predict the structure of a peptide that can be attached to the nanoparticles surface and promote an active targeting against the cells of interest.
3. Physic-chemical characterization of the formulation: The designed formulation was characterized in terms of chemical interactions, morphology, viscosity, thermo-responsiveness, and rheologic behavior.
4. Assess the biocompatibility and efficacy of the formulations in co-cultured and 3D in vitro models: Despite this part was not planned for the first year of grant, the biocompatibility of the designed formulation was tested in 2D healthy fibroblasts models, and preliminary results of in vitro efficacy were also performed in a 2D ovarian cancer cell model. Moreover, the optimization of the cell culture conditions for the growth of ovarian cancer 3D spheroids was also initiated.
6. Study the scale-up and stability of the formulation (e.g. sterilization, lyophilization, storage, etc.), as well as its possible translatability: The tasks associated with this objective were scheduled for the third year of the grant; however, it was tested the possibility to lyophilize the HG formulation.
7. Dissemination of the project results with the scientific community, citizenship, and potential stakeholders. The tasks associated with this objective were scheduled for the entire period of the grant. During the first year, some dissemination actions of the work were performed in scientific conferences and also in actions for citizenship.
The main achievements in terms of project results are:
Obtention of a hydrogel formulated using biocompatible, biodegradable, and natural polymers instead of the generally used and well-established synthetic polymers, with ideal physic-chemical and rheologic features, that demonstrated to be safe in vitro and able to slowly release the cargo during a long period of time (higher than 2 weeks). It was also possible to observe that the doses released are active in two different ovarian cancer cell lines.
It was also demonstrated that the in silico tools, can be very useful in this type of work, giving important clues for the perfect formulations design, with a reduction in resources and experimental work required.
In parallel, it was studied the possibility of decorate the surface of the PM with anti-CD133 moieties in order to promote an active targeting against the CD133 overexpressing cells, known by their higher resistance and malignancy. The internalization of CD133-functionalized versus non-functionalized particles in three different ovarian cancer cell lines, demonstrated the importance of this targeting, since in all the cell lines the internalization was higher for the functionalized particles. These results open a door to a more precise and improved treatments for advanced stages of ovarian cancer. In this sense, new peptides against CD133 were designed also with the help of in silico tools that could be used not only to target CD133 overexpressing ovarian cancer cells, but also to other tumors that present overexpression of this receptor.
Taking into account that only the first year of the project was executed during the time-course of this grant, a final formulation preclinical validated was not yet achieved. However, important preliminary results, including preliminary data on in vitro biological efficacy that was only initially planned for the second and third year of the project, where obtained and a promising formulation was achieved. Further research is required to validate the proposed system and demonstrate its feasibility as a depot system for the localized sustained release of chemotherapy to treat ovarian cancer.
Obtention of a hydrogel formulated using biocompatible, biodegradable, and natural polymers instead of the generally used and well-established synthetic polymers, with ideal physic-chemical and rheologic features, that demonstrated to be safe in vitro and able to slowly release the cargo during a long period of time (higher than 2 weeks). It was also possible to observe that the doses released are active in two different ovarian cancer cell lines.
It was also demonstrated that the in silico tools, can be very useful in this type of work, giving important clues for the perfect formulations design, with a reduction in resources and experimental work required.
In parallel, it was studied the possibility of decorate the surface of the PM with anti-CD133 moieties in order to promote an active targeting against the CD133 overexpressing cells, known by their higher resistance and malignancy. The internalization of CD133-functionalized versus non-functionalized particles in three different ovarian cancer cell lines, demonstrated the importance of this targeting, since in all the cell lines the internalization was higher for the functionalized particles. These results open a door to a more precise and improved treatments for advanced stages of ovarian cancer. In this sense, new peptides against CD133 were designed also with the help of in silico tools that could be used not only to target CD133 overexpressing ovarian cancer cells, but also to other tumors that present overexpression of this receptor.
Taking into account that only the first year of the project was executed during the time-course of this grant, a final formulation preclinical validated was not yet achieved. However, important preliminary results, including preliminary data on in vitro biological efficacy that was only initially planned for the second and third year of the project, where obtained and a promising formulation was achieved. Further research is required to validate the proposed system and demonstrate its feasibility as a depot system for the localized sustained release of chemotherapy to treat ovarian cancer.