Synergistic inhibition of the Hedgehog pathway by newly designed Smo and Gli antagonists bearing the isoflavone scaffold

Aberrant activation of the Hedgehog (Hh) pathway is responsible for the onset and progression of several malignancies. Small molecules able to block the pathway at the upstream receptor Smoothened (Smo) or the downstream effector Gli1 have thus emerged recently as valuable anticancer agents. Here, we have designed, synthesized, and tested new Hh inhibitors taking advantage by the highly versatile and privileged isoflavone scaffold. The introduction of specific substitutions on the isoflavone's ring B allowed the identification of molecules targeting preferentially Smo or Gli1. Biological assays coupled with molecular modeling corroborated the design strategy, and provided new insights into the mechanism of action of these molecules. The combined administration of two different isoflavones behaving as Smo and Gli antagonists, respectively, in primary medulloblastoma (MB) cells highlighted the synergistic effects of these agents, thus paving the way to further and innovative strategies for the pharmacological inhibition of Hh signaling

Italian guidelines for the use of antiretroviral agents and the diagnostic-clinical management of HIV-1 infected persons. Update 2016.

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Rate and Predictors of Self-Chosen Drug Discontinuations in Highly Active Antiretroviral Therapy-Treated HIV-Positive Individuals

Abstract Despite the clinical benefits of highly active antiretroviral therapy (HAART), sustained treatment remains a great challenge for HIV-infected people. The rate, consequences, and correlates of self-elected treatment interruptions (TI) are not known. The objectives of the study were to assess the rate of patient-elected TI in a cohort of HIVinfected people taking HAART, to evaluate whether patient-elected TI is correlated with suboptimal nonadherence, and to identify the predictors of self-chosen HAART interruptions. Using a Web-based cross-sectional survey beginning in January 2006 primary outcomes were: (1) reports of having asked their physician to interrupt the current regimen (AskDisc) and (2) reports of at least one interruption of a minimum of 1 day of any of the drugs included in the regimen (INTERR). Three hundred fifty-nine people were enrolled; 296 were taking HAART. Twenty-three percent self-reported suboptimal adherence, 45% reported AskDisc, and 25% INTERR. Forty percent of people reporting INTERR self-reported suboptimal adherence. As expected, AskDisc and INTERR were correlated with suboptimal adherence. The AskDisc group had higher CD4 cell counts and HIV RNA, more symptoms, and took more convenient regimens. The INTERR group had higher HIV RNA, were more likely to smoke, seek more information on HIV=AIDS, and less likely to take non-nucleoside reverse transcriptase inhibitors (NNRTIs). The rate of self-chosen TI was high and often related to suboptimal adherence. These findings may help clinicians to better monitor patients, and identify patients for targeted counseling

Long-acting agents for HIV infection: biological aspects, role in treatment and prevention, and patient's perspective

Current cART regimens are highly potent and well tolerated, but long-term toxicities, drug-drug interactions, lifetime costs and scarce option for multiclass failed patients could limit the efficacy of treatment itself. Long-acting formulations of antiretrovirals, which could potentially replace daily tablets, have been developed and are under investigation for prevention and treatment of HIV infection. Cabotegravir and rilpivirine represent the first drugs studied in this context. The aim of this review is to summarize the biological bases, the available information on completed and ongoing clinical trials and the potential development of long-acting regimens for the treatment and prevention of HIV infection

Compounds for use in the treatment of brain diseases

The present invention relates to a compound of formula (1): (1) or an analogue thereof or a pharmaceutically acceptable salt thereof, for use in the treatment of a brain tumor, wherein said compound is administered intranasally and relative pharmaceutical compositions

Discovery, characterization and potential roles of a novel NF-YAx splice variant in human neuroblastoma

Background: Identification of novel cancer-associated splice variants is of potential diagnostic, prognostic and therapeutic importance. NF-Y transcription factor is comprised of NF-YA, NF-YB and NF-YC subunits, binds inverted CCAAT-boxes in ≈70% of gene promoters, regulates > 1000 cancer-associated genes and proteins involved in proliferation, staminality, differentiation, apoptosis, metabolism and is subject to component alternative splicing. RT-PCR evaluation of alternative NF-YA splicing in primary human neuroblastomas (NBs), led to discovery of a novel NF-YAx splice variant, also expressed during mouse embryo development and induced by doxorubicin in NB cells. Here, we report the discovery and characterisation of NF-YAx and discus its potential roles in NB. Methods: NF-YAx cDNA was RT-PCR-cloned from a stage 3 NB (provided by the Italian Association of Haematology and Paediatric Oncology, Genova, IT), sequenced and expressed as a protein using standard methods and compared to known fully-spliced NF-YAl and exon B-skipped NF-YAs isoforms in: EMSAs for capacity to form NF-Y complexes; by co-transfection, co-immunoprecipitation and Western blotting for capacity to bind Sp1; by IF for localisation; in AO/EtBr cell-death and colony formation assays for relative cytotoxicity, and by siRNA knockdown, use of inhibitors and Western blotting for potential mechanisms of action. Stable SH-SY5Y transfectants of all three NF-YA isoforms were also propagated and compared by RT-PCR and Western blotting for differences in cell-death and stem cell (SC)-associated gene expression, in cell-death assays for sensitivity to doxorubicin and in in vitro proliferation, substrate-independent growth and in vivo tumour xenograft assays for differences in growth and tumourigenic capacity. Results: NF-YAx was characterized as a novel variant with NF-YA exons B, D and partial F skipping, detected in 20% of NF-YA positive NBs, was the exclusive isoform in a stage 3 NB, expressed in mouse stage E11.5-14 embryos and induced by doxorubicin in SH-SY5Y NB cells. The NF-YAx protein exhibited nuclear localisation, competed with other isoforms in CCAAT box-binding NF-Y complexes but, in contrast to other isoforms, did not bind Sp1. NF-YAx expression in neural-related progenitor and NB cells repressed Bmi1 expression, induced KIF1Bβ expression and promoted KIF1Bβ-dependent necroptosis but in NB cells also selected tumourigenic, doxorubicin-resistant, CSC-like sub-populations, resistant to NF-YAx cytotoxicity. Conclusions: The discovery of NF-YAx in NBs, its expression in mouse embryos and induction by doxorubicin in NB cells, unveils a novel NF-YA splice mechanism and variant, regulated by and involved in development, genotoxic-stress and NB. NF-YAx substitution of other isoforms in NF-Y complexes and loss of capacity to bind Sp1, characterises this novel isoform as a functional modifier of NF-Y and its promotion of KIF1Bβ-dependent neural-lineage progenitor and NB cell necroptosis, association with doxorubicin-induced necroptosis and expression in mouse embryos coinciding with KIF1Bβ-dependent sympathetic neuroblast-culling, confirm a cytotoxic function and potential role in suppressing NB initiation. On the other hand, the in vitro selection of CSC-like NB subpopulations resistant to NF-YAx cytotoxicity not only helps to explain high-level exclusive NF-YAx expression in a stage 3 NB but also supports a role for NF-YAx in disease progression and identifies a potential doxorubicin-inducible mechanism for post-therapeutic relapse