Periodic Reporting for period 1 - BOOMCHEMBAGS (Towards spatially-controlled, bioorthogonal, CuAAC-mediated assembly of targeted oncoprotein degraders)
Période du rapport: 2021-06-01 au 2023-05-31
The use of chemotherapeutical drugs is the most extended approach for cancer treatment due to their power to destroy cancer cells. However, most chemotherapists have the counterpart of an inherent toxicity for the rest of the body. The therapeutic strategies based on Bioorthogonal Chemistry aim to substantially reducing or even eliminating the undesired side effects of powerful pharmaceuticals by the bioorthogonal activation of the non-toxic prodrugs at the action place. The most extended technology for this purpose is based on the implantation at the action place of micro-devices consisting in micro-particles made of an artificial biocompatible polymer containing palladium or gold nanoparticles. Next, a bioorthogonal prodrug is introduced in the body not causing any effect until it reaches the bioorthogonal devices, where the metal nanoparticles catalyze the uncaging of the prodrug into the drug exclusively where the implants are. Although the therapeutic potential of focal bioorthogonally-controlled chemotherapy is vast, there are some issues associated to the catalytical devices that need to be addressed before translating this technology to the clinic. The matrix, although biocompatible, is not biodegradable, which prevents removal of the implant after therapy. Additionally, the direct contact between the metal’s surface and the proteins from the biological milieu, causes the nanoparticles’ coating reducing their catalytical capacity progressively. The research program of BOOMCHEMBAGS originated to fill this gap in the state-of-the-art by developing biocompatible and biodegradable devices designed to protect the surface of bioorthogonal catalyst based on metallic nanoparticles from direct contact with proteins from the biological milieu. To achieve this overall objective, the following specific objectives were planned:
O1. To develop biocompatible devices to contain catalytically-active Pd nanoparticles (PdNPs) protected from the direct contact with proteins and at the same time, reachable for small molecule substrate.
O2. To validate the functional properties of the Pd-devices under physiological conditions.
O3. To develop new prodrug/s to be bioorthogonally activated by the Pd-devices.
O4. To validate the new therapeutic strategy in biological experiments.
The work carried out during the implantation of BOOMCHEMBAGS project resulted in the successful achievement of the objectives proposed and more. A novel therapeutic strategy has been developed based on the activation of bioorthogonal prodrugs by catalytic devices made of the entrapment of PdNPs within microporous crystals made of natural occurring biopolymers (PdNPs@BpolCrystals). The devices allow the flow of small molecules such as prodrugs and drugs, but restrain the penetration of big molecules such as serum proteins. In this way, prodrugs can reach the PdNPs entrapped in the crystals, undergo the bioorthogonal uncaging reaction, and then, the drug is released exerting its actions. At the same time, the nanoparticles are fully isolated from the surrounding proteins, so their catalytic performance is maintained after several catalytic cycles. The unprecedented characteristics of the devices developed, solve the most crucial unmet requirements of previous metal-nanoparticles-based bioorthogonal devices. Additionally, beyond the original objectives, it was observed that the rate of the reaction catalysed by the new devices can be notably accelerated by irradiating them with harmless near infrared (NIR) radiation. This kind of radiation is known to be able to penetrate biological tissue triggering processes within the body by remote action. Together, the development of the new catalytic devices and prodrugs activable by them achieved in BOOMCHEMBAGS, bring this kind of therapeutic strategies closer to becoming a real treatment for patients treated with otherwise toxic drugs and will mean a great advance in the improvement of their quality of life.
O1. To develop biocompatible devices to contain catalytically-active Pd nanoparticles (PdNPs) protected from the direct contact with proteins and at the same time, reachable for small molecule substrate.
O2. To validate the functional properties of the Pd-devices under physiological conditions.
O3. To develop new prodrug/s to be bioorthogonally activated by the Pd-devices.
O4. To validate the new therapeutic strategy in biological experiments.
The work carried out during the implantation of BOOMCHEMBAGS project resulted in the successful achievement of the objectives proposed and more. A novel therapeutic strategy has been developed based on the activation of bioorthogonal prodrugs by catalytic devices made of the entrapment of PdNPs within microporous crystals made of natural occurring biopolymers (PdNPs@BpolCrystals). The devices allow the flow of small molecules such as prodrugs and drugs, but restrain the penetration of big molecules such as serum proteins. In this way, prodrugs can reach the PdNPs entrapped in the crystals, undergo the bioorthogonal uncaging reaction, and then, the drug is released exerting its actions. At the same time, the nanoparticles are fully isolated from the surrounding proteins, so their catalytic performance is maintained after several catalytic cycles. The unprecedented characteristics of the devices developed, solve the most crucial unmet requirements of previous metal-nanoparticles-based bioorthogonal devices. Additionally, beyond the original objectives, it was observed that the rate of the reaction catalysed by the new devices can be notably accelerated by irradiating them with harmless near infrared (NIR) radiation. This kind of radiation is known to be able to penetrate biological tissue triggering processes within the body by remote action. Together, the development of the new catalytic devices and prodrugs activable by them achieved in BOOMCHEMBAGS, bring this kind of therapeutic strategies closer to becoming a real treatment for patients treated with otherwise toxic drugs and will mean a great advance in the improvement of their quality of life.
The objectives of the project were addressed via three scientific Work Packages (WPs):
[WP1] Preparation & functional testing of Pd-based devices. A strategy based on the entrapment of PdNPs within biopolymer crystals was attempted, demonstrating being suitable to address all the required characteristics.. By measuring the uncaging of bioorthogonally protected pro-fluorophores, the catalytic performance of the PdNPs@BpolCrystals was proven and optimised. The fluorimetric method was employed to observe that their catalytical performance is not reduced after several cycles of bioorthogonal reactions. This unprecedented demonstrates that the strategy of entrapping PdNPs inside crystals is an excellent method to protect them while preserving their catalytic performance. In addition, an effective acceleration of the catalysed reactions when NIR laser was also observed. As far as we are aware, this is the first example of metal nanoparticles-catalysed bioorthogonal reaction remotely stimulated by NIR laser.
[WP2] Development of bioactive prodrugs activatable by Pd-based devices.
A drug that is known to be able to trigger the cell differentiation of stem cells into healthy tissue for tissue regeneration of patients treated with radiotherapy, was used as target. Currently, this drug cannot be employed due to its high toxicity. A bioorthogonal non-toxic derivative was synthetized. Furtherly, an effective uncaging of the prodrug by the devices was tested and proven. In addition, the strategy was successfully applied to other kind of drugs.
[WP3] Strategy validation: Bioorthogonal drug activation under biological conditions.
The validation of the strategy was carried out by an ex-vivo study in which excellent results regarding the safety of the devices have been obtained. The experiments to observe the effectiveness of the therapy are currently ongoing and excellent results are being obtained.
For exploitation purposes, both the new catalytic devices and the therapeutic strategy developed, are considered for intellectual protection. The results of the project were disseminated in two international scientific meetings and in two manuscripts currently awaiting final results.
[WP1] Preparation & functional testing of Pd-based devices. A strategy based on the entrapment of PdNPs within biopolymer crystals was attempted, demonstrating being suitable to address all the required characteristics.. By measuring the uncaging of bioorthogonally protected pro-fluorophores, the catalytic performance of the PdNPs@BpolCrystals was proven and optimised. The fluorimetric method was employed to observe that their catalytical performance is not reduced after several cycles of bioorthogonal reactions. This unprecedented demonstrates that the strategy of entrapping PdNPs inside crystals is an excellent method to protect them while preserving their catalytic performance. In addition, an effective acceleration of the catalysed reactions when NIR laser was also observed. As far as we are aware, this is the first example of metal nanoparticles-catalysed bioorthogonal reaction remotely stimulated by NIR laser.
[WP2] Development of bioactive prodrugs activatable by Pd-based devices.
A drug that is known to be able to trigger the cell differentiation of stem cells into healthy tissue for tissue regeneration of patients treated with radiotherapy, was used as target. Currently, this drug cannot be employed due to its high toxicity. A bioorthogonal non-toxic derivative was synthetized. Furtherly, an effective uncaging of the prodrug by the devices was tested and proven. In addition, the strategy was successfully applied to other kind of drugs.
[WP3] Strategy validation: Bioorthogonal drug activation under biological conditions.
The validation of the strategy was carried out by an ex-vivo study in which excellent results regarding the safety of the devices have been obtained. The experiments to observe the effectiveness of the therapy are currently ongoing and excellent results are being obtained.
For exploitation purposes, both the new catalytic devices and the therapeutic strategy developed, are considered for intellectual protection. The results of the project were disseminated in two international scientific meetings and in two manuscripts currently awaiting final results.
The therapeutic strategy developed in BOOMCHEMBAGS, solves the most crucial unmet requirements of previous bioorthogonal catalytic devices that keep them out of clinic. The advances made, bring this kind of therapeutic strategies closer to becoming a real treatment for patients treated with otherwise toxic drugs. The therapeutic strategy developed goes beyond the treatment of cancer which was the target of most of the previous bioorthogonal-metallic therapeutic approaches. The development of new prodrugs from non-chemotherapy drugs and their effective uncaging using the PdNPs@BpolCrystals, set the ground for the application of the technology developed to a panoply of treatments. In addition, the unprecedent observation of the acceleration of reactions catalysed by PdNPs@BpolCrystals when irradiated with NIR light, opens the door to the development of bioorthogonal therapeutic strategies in which the drug release can be controlled remotely using non-harmful radiation. This kind of remote control over a bioorthogonal process is totally new and is expected to have great impact on the development of novel treatment, but also on other research fields such as technology and industry.