Consensus clustering methodology to improve molecular stratification of non-small cell lung cancer.

Recent advances in machine learning research, combined with the reduced sequencing costs enabled by modern next-generation sequencing, paved the way to the implementation of precision medicine through routine multi-omics molecular profiling of tumours. Thus, there is an emerging need of reliable models exploiting such data to retrieve clinically useful information. Here, we introduce an original consensus clustering approach, overcoming the intrinsic instability of common clustering methods based on molecular data. This approach is applied to the case of non-small cell lung cancer (NSCLC), integrating data of an ongoing clinical study (PROMOLE) with those made available by The Cancer Genome Atlas, to define a molecular-based stratification of the patients beyond, but still preserving, histological subtyping. The resulting subgroups are biologically characterized by well-defined mutational and gene-expression profiles and are significantly related to disease-free survival (DFS). Interestingly, it was observed that (1) cluster B, characterized by a short DFS, is enriched in KEAP1 and SKP2 mutations, that makes it an ideal candidate for further studies with inhibitors, and (2) over- and under-representation of inflammation and immune systems pathways in squamous-cell carcinomas subgroups could be potentially exploited to stratify patients treated with immunotherapy.

Synthesis of 1,2,4-triazole derivatives: binding properties on endothelin receptors.

In the present study we describe the synthesis of a new series of 1,2,4-triazoles: [3-(arylmethyl)thio-5-aryl-4H-[1,2,4]triazol-4-yl]acetic acids 5a-g, [5-(arylmethyl)thio-3-aryl-1H-[1,2,4]triazol-1-yl]acetic acids 8a-d, and [3-(aryl-methyl)thio-5-aryl-1H-[1,2,4]triazol-1-yl] acetic acids 9a-d. These compounds were tested in binding assays to evaluate their ability as ligands for human ET(A) and ET(B) receptors stably expressed in CHO cells; some of the tested compounds showed affinity in the micromolar range.

Exploratory Analysis on Overall Survival after Either Surgery or Stereotactic Radiotherapy for Lung Oligometastases from Colorectal Cancer.

AIMS: Lung metastasectomy and, more recently, stereotactic body radiotherapy (SBRT), are frequently proposed to stage IV oligometastatic colorectal cancer (CRC) patients. In the absence of a randomised comparison between the two treatments, we aimed to retrospectively explore the effect on overall survival and progression-free survival (PFS) in a comparative cohort study. MATERIALS AND METHODS: We included patients who consecutively underwent surgery (n = 142) or SBRT (n = 28) as first local therapy at the time of lung progression, between 2005 and 2012. Both overall survival and PFS functions according to treatment were calculated using the Kaplan-Meier method and compared using the Log-rank test. The effect of treatment on overall survival and PFS was estimated by Cox models using different adjustment methods. RESULTS: Patients receiving SBRT were older and were treated more recently, whereas the two cohorts were similar for most baseline prognostic factors. Overall survival at 1 and 2 years was 0.89 and 0.77 for SBRT and 0.96 and 0.82 for surgery (P = 0.134), respectively. Multivariable analyses did not highlight a clear treatment effect on overall survival (adjusted hazard ratioSBRT versus surgery = 1.71; 95% confidence interval 0.82-3.54; P = 0.149) and even smaller differences using the inverse probability treatment weighting method (hazard ratioSBRT versus surgery = 1.28, 95% confidence interval 0.58-2.82; P = 0.547). The results of PFS were unreliable because different follow-up protocols were applied in the two cohorts. CONCLUSION: With limitations consisting in the retrospective observational design and different sample sizes, the results of this explorative analysis indicate that overall survival probability after SBRT is similar to surgery for the first 2 years from treatment. This finding supports the need for high-quality trials comparing different treatment modalities for lung oligometastases from CRC.

Anti-rhinovirus activity of 3-methylthio-5-aryl-4-isothiazolecarbonitrile derivatives.

A series of 3-methylthio-5-aryl-4-isothiazolecarbonitriles has been evaluated as anti rhinovirus agents against a panel of 17 representative human rhinovirus (HRV) serotypes, belonging to both A and B groups. No anti rhinovirus activity was detected for 3-methylthio-5-phenyl-4-isothiazolecarbonitrile (IS-2). Isothiazole derivatives with bulky substituents (O-Bn or O-But groups) on the para position of the phenyl ring were the most effective compounds of this series. In fact, a reduction in virus-induced cytopathogenicity was demonstrated for the O-Bn substituted IS-50 compound against the majority (88%) of the rhinoviruses tested, whereas the compound with an O-Ts group (IS-44) was found to be a specific inhibitor of group B serotypes, exhibiting the lowest IC(50) against HRVs type 2, 85 and 89. Our studies on the mechanism of action of IS-44 demonstrated that it prevents the thermal inactivation of HRV 2 infectivity, probably due to a conformational shift in the viral capsid and a decrease in affinity for the cellular receptor, resulting in an inhibition of attachment of the virions.

In vitro antiviral activity of four isothiazole derivatives against poliovirus type 1.

The in vitro effects of four isothiazoles [5,5'-diphenyl-3,3'-diisothiazole disulfide, 5-phenyl-3-mercapto-isothiazole, 5,5'-(4-chlorophenyl)-3,3'- diisothiazole disulfide, and 5-(4-chlorophenyl)-3-mercapto-isothiazole] on poliovirus type 1 were studied. The derivatives tested demonstrated remarkable viral inhibition, with a higher selectivity index than the previously studied iminodithiole precursors. Under one-step growth conditions, all the isothiazole derivatives caused the greatest activity if added during or after (within 1 h) poliovirus adsorption. These data suggest interference with early events of viral replication. [5-3H]Uridine incorporation into RNA showed that the compounds tested reduced poliovirus RNA synthesis, which was completely shut off after 2 h of incubation and reduced by 50-60% after 4 h. Also, pretreatment of the cell cultures with the compounds for 24 h caused a substantial inhibition of viral replication. The data suggest that the four isothiazole derivatives may have a multi-step antiviral mode of action different from their iminodithiole precursors.

Inhibitory effects of 3-imino-5-phenyl-3H-1,2-dithiole on poliovirus type 1 replication in vitro.

The effect of 3-imino-5-phenyl-3H-1,2-dithiole (PDTI) on different steps of the replicative cycle of poliovirus type 1 in HEp-2 cells was studied. This compound inhibited the replication of poliovirus type 1 as shown by cytopathic effect and virus yield reduction. This inhibitory action was not due to a virucidal effect, nor did the cells to have been pretreated. Under one-step growth conditions 3-imino-5-phenyl-3H-1,2-dithiole caused the greatest inhibition if added within 1 h after poliovirus adsorption. [5-3H]uridine incorporation into RNA showed that PDTI reduced poliovirus RNA synthesis. In fact, in the presence of PDTI viral RNA synthesis was shut off completely at 2 h post infection, and at 4 h post infection viral RNA synthesis was reduced by 50%. The compound may have an inhibitory effect on the early transcriptional and/or replicative functions of the poliovirus genome.

Synthesis of new 3-methylthio-5-aryl-4-isothiazolecarbonitriles with broad antiviral spectrum.

The isothiazole derivative 3-methylthio-5-(4-OBn-phenyl)-4-isothiazolecarbonitrile, coded IS-50, which in previous studies had exhibited a broad antipicornavirus spectrum of action, was selected as the model for the synthesis of a new series of 3-methylthio-5-aryl-4-isothiazolecarbonitriles. These compounds were prepared in good yield (from 66 to 82%) by alkylation of 3-methylthio-5-(4-hydroxyphenyl)-4-isothiazolecarbonitrile with suitable bromides in the presence of acetone; only the 4-cyanophenoxy derivatives were obtained in a yield of less than 30%. All the compounds were screened against a panel of 17 representative human rhinovirus (HRV) serotypes belonging to both A and B groups, enteroviruses polio 1, ECHO 9 and Coxsackie B1, cardiovirus EMC, measles virus, and herpes simplex virus type 1 (HSV-1). Our results demonstrate that HRV 86 (group A) and HRVs 39 and 89 (group B) are the rhinovirus serotypes more susceptible to the action of these compounds. Isothiazole derivatives with a longer intermediate alkyl chain exhibited good activity against polio 1 and ECHO 9. The compound bearing a butyl group between the two phenoxy rings showed the lowest IC(50) against Coxsackie B1 and measles viruses. No activity against HSV-1 was detected with any of the compounds screened.

Novel inhibitors of neuronal nitric oxide synthase.

Selective inhibitors of neuronal nitric oxide synthase (nNOS), which are devoid of any effect on the endothelial isoform (eNOS), may be required for the treatment of some neurological disorders. In our search for novel nNOS inhibitors, we recently described some 1-[(Aryloxy)ethyl]-1H-imidazoles as interesting molecules for their selectivity for nNOS against eNOS. This work reports a new series of 1-[(Aryloxy)alkyl]-1H-imidazoles in which a longer methylene chain is present between the imidazole and the phenol part of molecule. Some of these molecules were found to be more potent nNOS inhibitors than the parent ethylenic compounds, although this increase in potency resulted in a partial loss of selectivity. The most interesting compound was investigated to establish its mechanism of action and was found to interact with the tetrahydrobiopterin (BH(4)) binding site of nNOS, without interference with any other cofactors or substrate binding sites.

Synthesis and antifungal activity of pyrido[3',2':4,5]thieno[3,2-d]- 1,2,3-triazine derivatives.

The antimicrobial activity of some pyrido[3',2':4,5]thieno[3,2-d]- 1,2,3-triazine derivatives has been studied. Some compounds proved effective against microorganisms in vitro, compounds 3a and 3c in particular exhibited antifungal activity, comparable to MCZ, against hyphomycetes.

Synthesis of substituted [1]benzothieno[2,3-b]pyrazines and their ability to antagonize KCl induced contractions.

A series of substituted [1]benzothieno [2,3-b]pyrazines, structurally related to caroverine, was synthesized. Some of these compounds showed an appreciable inhibition towards KCl induced contractions on isolated rat aortic rings, and a lower potency towards negative inotropic activity tested on isolated guinea pig atrium.

Synthesis and pharmacological properties of 2-nitro-3-substituted-amino benzo[b]thiophenes and 1-substituted benzothieno[2,3-d]imidazoles.

The synthesis of 1-substituted benzothieno[2,3-d]imidazoles and their most suitable synthesis precursor 2-nitro-3-substituted-amino benzo[b]thiophenes is reported. Most of these latter compounds show analgesic and antiinflammatory activities sometimes comparable to those of phenylbutazone with less toxicity and gastric lesivity.

Synthesis and pharmacological properties of 3-alkylthio derivatives of isothiazolothieno-1,2,3-triazine.

Some 3-alkylthio- derivatives of 7-chloroisothiazolothieno-1,2,3-triazine and of isothiazolothieno-1,2,3-triazin-7(4H)one were prepared. The synthesized compounds were subjected to pharmacological screening for antiallergic, antiinflammatory and analgesic activity as well as to a microbiological test in order to evaluate potential antifungal activity.

Regulation of cerebral kynurenine and 5-hydroxyindole pathways during tryptophan loading.

Cerebral tryptophan metabolism was studied in rat brain slices. The results show that serotonin production was inhibited at high levels of tryptophan or 5-oxo-L-prolyltryptophan. In contrast, kynurenine formation showed a dose dependent increase at the various concentrations of tryptophan or 5-oxo-L-prolyltryptophan. Measurements of the activities of tryptophan hydroxylase (TH) and indoleamine 2,3-dioxygenase (IDOase) in crude brain homogenates showed that serotonin formation was linear in the presence of dithiothreitol, whereas kynurenine production was inhibited in the presence of dithiothreitol or superoxide dismutase. These results suggest an inverse relationship in the regulation of 5-hydroxyindole and kynurenine pathways. The former being inhibited in the presence of high tryptophan concentration while the latter is enhanced. Furthermore, a high intracellular thiol-disulphide ratio appears to favour serotonin formation, whereas a highly reducing environment decreases kynurenine production.

Synthesis of 3-aminothieno-(2,3-b)pyridine derivatives. II. / Sintesi di derivati della 3-aminotieno (2,3-b) piridina. Nota II

The 3-cyanopyridin-2-thiol and its alkyl derivatives give, with halogen derivatives having an electron attracting group in alpha-position, the 3-aminothieno (2,3-b) pyridine derivatives in high yields. The behaviour of 2-carboethoxy-3-amino derivates and its products of hydrolysis and acetylation are studied.

Inhibition of neuronal nitric oxide synthase by N-phenacyl imidazoles.

Nitric oxide (NO) mediates a series of physiological processes, including regulation of vascular tone, macrofage-mediated neurotoxicity, platelet aggregation, learning and long-term potentiation, and neuronal transmission. Although NO mediates several physiological functions, overproduction of NO can be detrimental and play multiple roles in several pathological diseases. Accordingly, more potent inhibitors, more selective for neuronal nitric oxide synthase (nNOS) than endothelial NOS (eNOS) or inducible NOS (iNOS), could be useful in the treatment of cerebral ischemia and other neurodegenerative diseases. We recently described the synthesis of a series of imidazole derivatives. Among them N-(4-nitrophenacyl) imidazole (A) and N-(4-nitrophenacyl)-2-methyl-imidazole (B) were considered selective nNOS inhibitors. In the present study the action mechanism of compounds A and B was analyzed. Spectral changes observed in the presence of compound A indicate that this inhibitor exerts its effect without interaction with heme iron. Moreover compounds A and B, inhibit nNOS "noncompetitively" versus arginine, but "competitively" versus BH(4).

[Thioamide derivatives with gastro-protective activity]. / Derivati tioamidici ad attività gastroprotettiva.

A series of 3-aryl-prop-2-enthioamide derivatives has been synthesized. These compounds were evaluated for gastro-protective activity in the rat. It has been shown that the N,N-dimethylamino derivative was the most active compound in this series [Table I comp. (II)].

Synthesis and pharmacological activity of 3-substituted pyrido[3',2':4,5]thieno[3,2-d]pyrimidin-4(3H)-ones.

A series of 3-substituted pyridothienopyrimidin-4(3H)-ones has been synthesized from 2,7,9-trimethyl-4H-pyrido[3',2':4,5]thieno-[3,2-d] [ 1,3]oxazin-4-one. These compounds have been evaluated for analgesic, anti-inflammatory and antipyretic activities. The ulcerogenic effect of the compounds has been also studied.