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The pathogenesis of many cancers is characterized by mutations, overexpression and dysregulation of protein kinases. As a result, increasing attention has been directed towards the identification of novel kinase inhibitors for cancer therapy. The work performed here focuses on the synthesis of a series of pyrazolo[3,4-d]pyrimidine derivatives as inhibitors of the serine-threonine kinase SGK1, and the tyrosine kinases Fyn and Src. The first set of compounds are analogues of the in house SGK1 inhibitor SI113 which had previously demonstrated anti-cancer activity. In fact SI113 resulted to be active on various cancer cell lines and in an in vivo hepatocellular carcinoma model. The new set of SI113 analogues are characterized by different anilines, ammines and a morpholine group in C4 and are decorated in C6 with polar chains, i.e. ethanolamine, diethanolamine, ethylene glycol and ethylenediamine. The double bond on the N1 side chain which is essential for the activity of compounds toward SGK1, was maintained. The structures of the second set of compounds, instead, are related to the in house Fyn inhibitor SI308. From previously studies on the first generation of Fyn inhibitors, SI308 was reported as the most potent compound, demonstrating both antiproliferative activity on cancer cell lines and the ability to inhibit protein Tau phosphorylation in a cellular model of Alzheimer?s disease. The new generation of SI308 related compounds present a methyl group at the C6 position and on C3 the phenyl ring is either unsubstituted or presents a methyl group in para position. Preliminary screening using enzyme activity assays demonstrates that some of the novel compounds are active and therefore suitable for further study using in vitro models. It is predicted that subsequent in vitro data will aid in the design of future compounds. Furthermore, previous data on another in house compound SI306, which is active on tyrosine kinase Src, reported promising results on an in vivo xenograft model of neuroblastoma. To further study this biological activity, SI306 was re- synthesized and additional experiments were performed on in vitro models. Testing of this compound on MYCN-amplified neuroblastoma cell lines HTLA-230 and SK-N-BE-2C further confirmed the activity of SI306 and provide increased support for the inhibition of Src as a valid approach for neuroblastoma treatment. Finally, further in vitro studies were performed on of the three previously cited in house pyrazolo[3,4-d]pyrimidines, SI306, SI308 and SI113, using patient derived glioblastoma multiforme (GBM) cell lines. This final set of work was undertaken during a visiting research fellowship period and performed in collaboration with the School of Pharmacy at the University of Nottingham (United Kingdom). Kinase inhibitor activity has been evaluated on series of patient derived GBM cell lines isolated from both the central tumor core (GCE28) and from the invasive margin of the tumor (GIN28 and GIN8). The use of such phenotypically relevant in vitro models represents an important step for GBM drug development and screening. The results gathered using these relevant cell models further demonstrate the anti-cancer activity of the pyrazolo[3,4-d]pyrimidines compounds. Moreover, investigating the compounds in combination with one another reveals that synergy can be achieved and this finding has additional implications for potentially overcoming GBM drug resistance. Additionally, to overcome the low water solubility of our pyrazolo[3,4-d]pyrimidines compounds, formulations of the lead compounds (SI306, SI308 and SI113) were prepared using miniaturized screening process based on inkjet printing technology6. The observed activity of our compounds in vitro taken together with their successful formulation highlight that our kinase inhibitors are attractive candidates for the treatment of GBM.

Fanconi anemia (FA) is a rare genetic disease characterized by congenital malformations, aplastic anemia and increased risk of developing malignancies. FA is genetically heterogeneous as it is caused by at least 17 different genes. Among these, FANCA, FANCC, and FANCG account for approximately 85% of the patients whereas the remaining genes are mutated in only a small percentage of cases. For this reason, the molecular diagnostic process is complex and not always extended to all the FA genes, preventing the characterization of individuals belonging to rare groups.The FA genes were analyzed using a next generation sequencing approach in two unrelated families.The analysis identified the same, c.484_485del, homozygous mutation of FANCF in both families. A careful examination of three electively aborted fetuses in one family and one affected girl in the other indicated an association of the FANCF loss-of-function mutation with a severe phenotype characterized by multiple malformations.The systematic use of next generation sequencing will allow the recognition of individuals from rare complementation groups, a better definition of their clinical phenotypes, and consequently, an appropriate genetic counseling.

Pyrazolo[3,4-d]pyrimidines represent an important class of heterocyclic compounds well-known for their anticancer activity exerted by the inhibition of eukaryotic protein kinases. Recently, pyrazolo[3,4-d]pyrimidines have become increasingly attractive for their potential antimicrobial properties. Here, we explored the activity of a library of in-house pyrazolo[3,4-d]pyrimidines, targeting human protein kinases, against Staphylococcus aureus and Escherichia coli and their interaction with ampicillin and kanamycin, representing important classes of clinically used antibiotics. Our results represent a first step towards the potential application of dual active pyrazolo[3,4-d]pyrimidine kinase inhibitors in the prevention and treatment of bacterial infections in cancer patients.

In judgment 32/2020 the Italian Constitutional Court has proclaimed, for the first time, the prohibition of ex post facto law applicable to the norms governing the execution of criminal sanctions, and especially to the norms concerning the so called ‘alternative measures to detention’. The present paper offers an overview of the Italian legal framework and previous case law regarding the prohibition of ex post facto law and alternative measures. It also analyzes the innovative conclusions expressed by judgment 32/2020. Finally, it stresses the most remarkable aspect of the judgment, which is the use that the Constitutional Court makes of the US Supreme Court’s case law to re-interpret Art. 25 (2) of the Italian Constitution. Con la sentenza n.32/2020 la Corte costituzionale italiana ha proclamato, per la prima volta, il divieto della legge ex post applicabile alle norme che regolano l'esecuzione delle sanzioni penali, e in particolare alle norme relative alle cosiddette "misure alternative alla detenzione" . Il presente documento offre una panoramica del quadro giuridico italiano e della precedente giurisprudenza relativa al divieto della legge ex post e delle misure alternative. Analizza inoltre le conclusioni innovative espresse dalla sentenza 32/2020. Infine, sottolinea l'aspetto più notevole della sentenza, che è l'uso che la Corte costituzionale fa della giurisprudenza della Corte suprema degli Stati Uniti per reinterpretare l'Arte. 25 (2) della Costituzione italiana., Roma Tre Law Review – 01/2020

Questa sezione del volume dedicata al rapporto tra sociologia, linguaggio e genere mira ad approfondire la riflessione sulle relazioni tra cambiamento socio-culturale ed evoluzione degli usi linguistici e comunicativi, partendo dall'idea che le parole possono, da un lato, marginalizzare ed emarginare; dall'altro lato, possono però diventare un efficace strumento della lotta alle disuguaglianze basate sul genere e sull'orientamento sessuale.