All histological scores were significantly decreased in the CRB0017 12 μg/knee group compared to vehicle, while administration of CRB0017 1.2 μg was associated with a trend to a decrease in the same parameters. Therefore, CRB0017 administered twice in 3 months could modify the course of OA in the STR/ort mouse, by delaying cartilage breakdown as assessed histologically. The procedure of blind scoring of the histological samples clearly showed that knee intra-articular administration of CRB0017, an anti-ADAMTS5 antibody, dose-dependently improved disease progression in a relevant animal model of OA.

Multivalency is widely used in nature in specific recognition processes. This paper describes an approach to multivalency in the pentameric 5-HT3 receptor, a ligand-gated ion channel, which constitutes an example of intrinsically multivalent biological receptors. Owing to the picomolar Ki value, TETRA-L represents an outstanding multivalent ligand for the neurotransmitter receptor.

An adaptive method for designing the infill pattern of 3D printed objects is proposed. In particular, new unit cells for metamaterials are designed in order to match prescribed mechanical specifications. To this aim, we resort to topology optimization at the microscale driven by an inverse homogenization to guarantee the desired properties at the macroscale. The whole procedure is additionally enriched with an anisotropic adaptive generation of the computational mesh. The proposed algorithm is first numerically verified both in a mono-and in a multi-objective context. Then, a mechanical validation and 3D manufacturing through fused-model-deposition are carried out to assess the feasibility of the proposed design workflow. 1 MOTIVATIONS Recently, the developing of innovative manufacturing technologies has involved the industry in numerous fields (e.g., automotive, aerospace, medical industry) [4, 23]. Among the different innovations, the Additive Manufacturing (AM) has revolutionized the way to think the production [2]. In fact, the AM has allowed the production of objects with complex geometries in a simple way, overcoming the constraints

Tolerance to opioid administration represents a serious medical alert in different chronic 28conditions. This study compares the effects of the imidazoline compounds 1, 2, and 3 on 29 morphine tolerance in an animal model of inflammatory pain in the rat. 1, 2, and 3 have been 30 selected in that, although bearing a common scaffold, preferentially bind to α 2 -adrenoceptors, 31 imidazoline I 2 receptors, or both systems, respectively. Such compounds have been tested in 32 vivo by measuring the paw withdrawal threshold to mechanical pressure after complete 33Freund's adjuvant injection. To determine the ligand levels in rat plasma, an HPLC-mass 34 spectrometry method has been developed. All the compounds significantly reduced the 35 induction of morphine tolerance, showing different potency and duration of action. Indeed, 36 the selective imidazoline I 2 receptor interaction (2) restored the analgesic response by 37 maintaining the same time-dependent profile observed after a single morphine administration. 38Differently, the selective α 2C -adrenoceptor activation (1) or the combination between α 2C -39 adrenoceptor activation and imidazoline I 2 receptor engagement (3) promoted a change in the 40 temporal profile of morphine analgesia by maintaining a mild but long lasting analgesic 41 effect. Interestingly, the kinetics of compounds in rat plasma supported the pharmacodynamic 42 data. Therefore, this study highlights that both peculiar biological profile and bioavailability 43 of such ligands complement each other to modulate the reduction of morphine tolerance. 44Based on these observations, 1-3 can be considered useful leads in the design of new drugs 45 able to turn off the undesired tolerance induced by opioids. 46 47

BACKGROUND: Currently there is no cure for glioblastoma (GBM), the most malignant primary brain cancer. Glioblastoma etiopathogenesis involves mutations and alterations of key signaling pathways including tyrosine kinases (TKs). Notably, aberrant activation of receptor tyrosine kinases (RTKs) such as EGFR (amplified in 40% of glioblastoma cases) and KDR represents one of the driving forces for cellular proliferation and angiogenesis. The ability to cross the blood brain barrier (BBB) and appropriate pharmacodynamics (target selectivity) and pharmacokinetics (brain penetration) are critical issues for the generation of potential drug candidates against glioblastoma. CR13626 is a novel brain penetrant small molecule able to inhibit different TKs relevant to the development of this cancer. The present study describes the in vitro and in vivo properties of CR13626 and its antitumor activity in a mouse model of glioblastoma. METHODS: We identified the TKs targeted by CR13626 against a panel of 173 kinase enzymes. The effects of CR13626 on cellular proliferation were assessed in different 2D human GBM cell lines (U87MG, U373, U87MG vIII) by ViaCount, and in U87MG 3D spheroids by CellTiter-Glo 3D viability assay. The antitumor activity in vivo was determined in an orthotopic mouse model of GBM based on the injection of U87MG-Luciferase cells in nude mice. Animals were orally treated with CR13626 (50 mg/kg/daily) for 10 days, starting on day 9 post-implantation. Measurement of bioluminescence (BLI) at day 19 (end of dosing) and during follow-up (days 26-33) allowed the monitoring of tumor progression. CR13626 pharmacokinetics and brain exposure were assessed by LC/MS/MS in plasma and brain homogenate tissues of CD1 and tumor-bearing nude mice. RESULTS: CR13626 potently inhibited tyrosine kinases relevant to GBM development, with IC50 values in the nanomolar range: FYN (69 nM), YES (3.6 nM), KDR (82 nM) and EGFR (6 nM). CR13626 reduced the growth of different human glioblastoma cell lines with GI50 values in the range 1-3 µM. In vivo, CR13626 treatment led to a time-dependent reduction of tumor growth, reaching 60% on the last BLI evaluation 33 days post-implantation (i.e. 15 days after the end of dosing). A 25% significant increase in the median survival of animals compared to the vehicle group was also observed. The antitumor effects of CR13626 were in line with the exposure of tumor-bearing mice to the compound. Pharmacokinetic studies in CD1 mice showed good oral bioavailability (72%) and brain penetration (brain/plasma ratio of 1.4) for CR13626. CONCLUSION: The ability of CR13626 to cross the BBB without being a substrate of efflux transporters that mediate tumor resistance, to simultaneously inhibit the activity of different TKs involved in GBM development, and to reduce tumor growth eventually leading to an increased survival of animals, warrant its further development as a drug candidate in glioblastoma. Citation Format: Chiara Galimberti, Tiziana Piepoli, Giuseppe M. Montagna, Silvia Zerbi, Ornella Letari, Roberto Artusi, Milena Colovic, Stefano Persiani, Gianfranco Caselli, Lucio C. Rovati. Efficacy of CR13626, a novel oral brain penetrant multi-kinase inhibitor, in a mouse model of glioblastoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4219.