New short strategy for the synthesis of the dibenz[b,f]oxepin scaffold.

In this report a short and efficient synthesis of the dibenz[b,f]oxepin framework through intramolecular SNAr and McMurry reactions is described. The diaryl ethers required for the McMurry reaction have been obtained in good yields under microwave-assisted conditions of the reaction of salicylaldehydes with fluorobenzaldehydes without catalysts. Application of an intramolecular McMurry reaction to the synthesized diarylethers using TiCl4/Zn in THF gave the target dibenzo[b,f]oxepin system in 53%-55% yields.

A one-pot sequence for the efficient synthesis of highly functionalized macrocarbocycles or bridged 2,8-dioxabicyclo[3.2.1]octanes from 1-nitrobicyclic compounds.

The reaction of 1-nitrobicyclo[n.3.1]alkane-(6 + n)ones with sodium borohydride followed by acidic workup led to ring opening via a one-pot sequence comprising the retro-Dieckmann-type opening of the α-nitroketone structural fragment, followed by aldehyde reduction and a final Nef reaction, leading to highly functionalized 12 to 14-membered carbocyclic ketones bearing three stereocenters, which are adjacent in some of the compounds. The reactions starting from 1-nitrobicyclo[9.3.1]pentadecan-15-ones could be adjusted to give macrocyclic 2,8-dioxabicyclo[3.2.1]octanes containing an additional bridge by diastereoselective formation of a third ring and a fourth stereocenter through acid-promoted intramolecular ketal formation. This is a very interesting ring system related to the core of the zaragozic acid family of natural products.

Domino reactions for the synthesis of bridged bicyclic frameworks: fast access to bicyclo[n.3.1]alkanes.

In the current bid for synthetic efficiency, multiple bond-forming transformations are becoming a key concept in organic synthesis. In this context, domino protocols have become increasingly common in the synthesis of bicyclic systems. This tutorial review aims at providing a short but authoritative overview of this topic, focusing on the preparation of bridged bicyclo[n.3.1]alkane systems, and is organized according to the number of rings created in the key domino process.

Multitarget Hybrid Fasudil Derivatives as a New Approach to the Potential Treatment of Amyotrophic Lateral Sclerosis.

Hybrid compounds containing structural fragments of the Rho kinase inhibitor fasudil and the NRF2 inducers caffeic and ferulic acids were designed with the aid of docking and molecular mechanics studies. Following the synthesis of the compounds using a peptide-coupling methodology, they were characterized for their ROCK2 inhibition, radical scavenging, effects on cell viability (MTT assay), and NRF2 induction (luciferase assay). One of the compounds (1d) was selected in view of its good multitarget profile and good tolerability. It was able to induce the NRF2 signature, promoting the expression of the antioxidant response enzymes HO-1 and NQO1, via a KEAP1-dependent mechanism. Analysis of mRNA and protein levels of the NRF2 pathway showed that 1d induced the NRF2 signature in control and SOD1-ALS lymphoblasts but not in sALS, where it was already increased in the basal state. These results show the therapeutic potential of this compound, especially for ALS patients with a SOD1 mutation.

Synthesis of benzo- and naphtho-fused bicyclo[n.3.1]alkane frameworks with a bridgehead nitrogen function by palladium-catalyzed intramolecular α'-arylation of α-nitroketones.

The C-alkylation of cyclic α-nitroketones with α-halobenzyl halides in the presence of DBU followed by a Pd-catalyzed intramolecular C-arylation afforded benzo-and naphtho-fused bicyclo[n.3.1]alkane derivatives (n = 3, 4, 5) in excellent overall yields for the two-step sequence. In some of the reactions starting from α-nitrocyclooctanone, the major products were fused indane derivatives arising from an intramolecular attack of an intermediate Pd species onto the carbonyl group, followed by elimination.

Red Beetroot's NMR-Based Metabolomics: Phytochemical Profile Related to Development Time and Production Year.

Red beetroot (RB) is a well-known health-promoting food consumed worldwide. RB is commonly used in food processing and manufacturing thanks to the high content of components that can also be employed as natural coloring agents. These bioactive molecules vary their concentration depending on beetroot seasonality, harvest time and climate conditions. The first objective of this study was to evaluate the variation of the RB phytochemical profile related to the root development during three different harvest times, using an 1H-NMR-based metabolomic approach. Changes of carbohydrates and secondary metabolite concentrations were observed from July to September. Secondly, we compared the metabolic profiles of the final processed beet juices in three different production years to observe the effect of climate conditions on the RB's final product metabotype. A PCA analysis performed on juice extracts showed that production years 2016 and 2017 were characterized by a high content of choline and betaine, while 2018 by a high content of amino acids and dopamine and a low content of inorganic nitrates. This study suggests that the harvest time and roots growth conditions could be used to modulate the RB phytochemical profile, according to the final requirements of use, food or coloring agent source.

Enzymatic-nonenzymatic cellular antioxidant defense systems response and immunohistochemical detection of MDMA, VMAT2, HSP70, and apoptosis as biomarkers for MDMA (Ecstasy) neurotoxicity.

3,4-Methylenedioxymethamphetamine (MDMA)-induced neurotoxicity leads to the formation of quinone metabolities and hydroxyl radicals and then to the production of reactive oxygen species (ROS). We evaluated the effect of a single dose of MDMA (20 mg/kg, i.p.) on the enzymatic and nonenzymatic cellular antioxidant defense system in different areas of rat brain in the early hours (<6 hr) of the administration itself, and we identified the morphological expressions of neurotoxicity induced by MDMA on the vulnerable brain areas in the first 24 hr. The acute administration of MDMA produces a decrease of reduced and oxidized glutathione ratio, and antioxidant enzyme activities were significantly reduced after 3 hr and after 6 hr in frontal cortex. Ascorbic acid levels strongly increased in striatum, hippocampus, and frontal cortex after 3 and 6 hr. High levels of malonaldehyde with respect to control were measured in striatum after 3 and 6 hr and in hippocampus and frontal cortex after 6 hr. An immunohistochemical investigation on the frontal, thalamic, hypothalamic, and striatal areas was performed. A strong positive reaction to the antivesicular monoamine transporter 2 was observed in the frontal section, in the basal ganglia and thalamus. Cortical positivity, located in the most superficial layer was revealed only for heat shock protein 70 after 24 hr.

The efficacy and tolerability of glucosamine sulfate in the treatment of knee osteoarthritis: A randomized, double-blind, placebo-controlled trial.

BACKGROUND: Osteoarthritis (OA) is the most common form of arthritis and is often associated with disability and impaired quality of life. OBJECTIVE: The aim of the study was to assess the efficacy and tolerability of glucosamine sulfate (GS) in the treatment of knee OA. METHODS: Consecutive outpatients affected by primary monolateral or bilateral knee OA were enrolled in this double-blind, double-dummy, prospective, randomized, placebo-controlled trial. One group received GS 1500 mg QD for 12 weeks, and the other group received placebo QD for 12 weeks. The treatment period was followed by a 12-week treatment-free observation phase. Each patient was examined at baseline and at weeks 4, 8, 12, 16, 20, and 24. The primary efficacy criteria were pain at rest and during movement, assessed on a visual analog scale (VAS) of 0 to 100 mm. The secondary criteria included the Western Ontario and McMaster Universities (WOMAC) index for total pain score (W-TPS), total stiffness score (W-TSS), and total physical function score (W-TPFS). VAS, W-TPS, W-TSS, and W-TPFS were evaluated at baseline and at weeks 4, 8, 12, 16, 20, and 24. Analgesic drug consumption (ie, acetaminophen or NSAIDs) was also assessed. RESULTS: Patient demographics were similar in the GS and placebo groups. Of 60 randomized patients (30 per group), 56 completed the study (28 treated with GS and 28 who received placebo). Statistically significant improvements in symptomatic knee OA were observed, as measured by differences in resting pain at weeks 8, 12, and 16 (all, P < 0.05 vs placebo) and in pain during movement at weeks 12 and 16 (both, P < 0.05). W-TPS was lower with GS than placebo at weeks 8, 12, and 16 (all, P < 0.01), and at week 20 (P < 0.05). W-TSS was also lower with GS than placebo at weeks 8, 12, 16, and 20 (all, P < 0.05). W-TPFS was lower with GS than placebo at weeks 8 (P < 0.05), 12 (P < 0.01), 16 (P < 0.05), and 20 (P < 0.05). Drug consumption was lower in the GS group than the placebo group at weeks 8, 12, 16, and 20 (all, P < 0.05). The incidence of adverse events was 36.7% with GS and 40.0% with placebo. CONCLUSIONS: GS 1500 mg QD PO for 12 weeks was associated with statistically significant reductions in pain and improvements in functioning, with decreased analgesic consumption, compared with baseline and placebo in these patients with knee OA. A carryover effect was detected after treatment ended.

Calcium bioavailability from a new calcium-fortified orange beverage, compared with milk, in healthy volunteers.

The recommended dietary calcium intake may be difficult to reach using dairy products only. This study aimed to evaluate the absorbability of calcium of a new orange beverage in comparison with that of milk. Ten healthy adults, 5 males and 5 females, were randomly divided to receive, under fasting conditions, an orange beverage enriched either with calcium or milk. Both the beverages had been labelled the previous evening with a 44Ca solution. After a two-month interval the subjects underwent a crossover of beverages. Total and ionized calcium, parathyroid (PTH) hormone, and 44Ca were evaluated. The 44Ca was measured by inductively coupled plasma spectrometry. The relative changes in calcium absorption from the two test drinks were similar (23.0 +/- 6.4% for milk and 20.9 +/- 9.1% for the orange beverage). No significant differences were found between the two test groups. Both milk and orange beverage determined a similar and significant (p < 0.001) decrease in serum PTH two hours after the beginning of the test (-22.1 +/- 14.0% for milk and -27.9 +/- 15.3% for orange beverage). We can conclude that the bioavailability of calcium from this new calcium-enriched orange beverage is similar to that of milk.

Chemo-immunotherapy of metastatic colorectal carcinoma with gemcitabine plus FOLFOX 4 followed by subcutaneous granulocyte macrophage colony-stimulating factor and interleukin-2 induces strong immunologic and antitumor activity in metastatic colon cancer patients.

PURPOSE: Tumor cell killing by anticancer drugs may be supported by their immuno- and pharmacologic effects. Chemotherapy is in fact able to (A) upregulate tumor-associated antigen expression, including carcinoembryonic antigen (CEA) or other target molecules such as thymidylate synthase (TS); and (B) downregulate tumor cell resistance to the death signals induced by tumor antigen-specific cytotoxic T lymphocytes. This provides the rationale for combining chemo- and immunotherapy. MATERIALS AND METHODS: We describe the results of a translational phase II trial designed to evaluate the toxicity, antitumor activity and immunologic effects of gemcitabine + FOLFOX-4 (oxaliplatin, fluorouracil, and folinic acid) polychemotherapy followed by the subcutaneous administration of granulocyte macrophage colony-stimulating factor and low-dose interleukin-2 in colorectal carcinoma patients. The study involved 29 patients (16 males and 13 females with a mean age of 69 years), 21 of whom had received a previous line of treatment, and 19 had liver involvement. RESULTS: The treatment was well tolerated and induced very high objective response (68.9%) and disease control rates (96.5%), with an average time to progression of 12.5 months. An immunologic study of peripheral blood mononuclear cells (PBMCs) taken from 20 patients showed an enhanced proliferative response to colon carcinoma antigen and a significant reduction in suppressive regulatory T lymphocytes (CD4+CD25T-reg+). A cytofluorimetric study of the PBMCs of five HLA-A(*)02.01+ patients who achieved an objective response showed an increased frequency of cytolytic T lymphocyte precursors specific for known CEA- and TS-derived epitopes. CONCLUSION: The results show that our regimen has strong immunologic and antitumor activity in colorectal cancer patients and deserves to be investigated in phase III trials.

Affinity of Iresine herbstii and Brugmansia arborea extracts on different cerebral receptors.

Iresine herbstii Hook. (Amaranthaceae) and Brugmansia arborea (L.) Lagerheim (Solanaceae) are used in the northern Peruvian Andes for magic-therapeutical purposes. The traditional healers use Iresine herbstii with the ritual aim to expel bad spirits from the body. Furthermore, Iresine herbstii was used in association with other plants, such as Trichocereus pachanoi Britt. et Rose, for divination, to diagnose diseases, and to take possession of another identity. Also, species of Brugmansia have been reported to be used during ritual practices for magical and curative purposes. Given the above evidence, the aim of the present study is to evaluate if the central effects of Iresine herbstii and Brugmansia arborea could be associated with interaction with SNC receptors. Two Iresine herbstii extracts (methanolic and aqueous) and one Brugmansia arborea aqueous extract were tested for in vitro affinity on 5-HT(1A), 5-HT(2A), 5-HT(2C), D1, D2, alpha(1), and alpha(2) receptors by radioligand binding assays. The biological materials for binding assay (cerebral cortex) were taken from male Sprague-Dawley rats. The extracts affinity for receptors is definite as inhibition percentage of radioligand/receptor binding and measured as the radioactivity of remaining complex radioligand/receptor. The data obtained for Iresine extracts have shown a low affinity for the 5-HT(1A) receptor and no affinity for 5-HT(2A) receptor. Otherwise the methanolic extract showed affinity for 5-HT(2C) receptor (IC(50): 34.78 microg/ml) and for D1 receptor (IC(50): 19.63 microg/ml), instead the Iresine aqueous extract displayed a lower affinity for D1 (48.3% at the maximum concentration tested) and a higher value of affinity for D2 receptors (IC(50): 32.08 microg/ml). The Brugmansia aqueous extract displayed affinity for D1 receptors (IC(50): 17.68 microg/ml), D2 receptors (IC(50): 15.95 microg/ml) and weak affinity for the serotoninergic receptors. None of the three extracts showed relevant affinity to the alpha(1), and alpha(2) receptors. The results of our experiments indicate that Iresine herbstii methanolic extract was able to interact with the central 5-HT(2C) and D1 receptors and Iresine herbstii aqueous extract showed affinity for D2 receptors, thus confirming their ritual use. Instead Brugmansia arborea was able to interact only with the central dopamine receptors tested. Parallel studies are currently in progress for evaluating the extracts affinity and active components towards these and other receptor types (GABAergic).

Binding studies for serotoninergic, dopaminergic and noradrenergic receptors of Valeriana adscendens Trel. extracts.

Valeriana adscendens Trel. (Valerianaceae) is a psychoactive plant usually used in magical-therapeutic rituals in traditional practices of the Northern Peruvian Andes. Previous studies have been carried out on extracts of aerial parts in order to validate its traditional use. The results indicated that Valeriana adscendens exerts important effects on the central nervous system. Aim of the present study is to evaluate if the effects on the central nervous system of Valeriana adscendens extracts can be associated with interaction with some CNS receptors. In this work we examined affinity and selectivity of two Valeriana adscendens extracts (methanolic and aqueous) towards 5-HT(1A), 5-HT(2A), 5-HT(2C) serotononergic, D(1) and D(2) dopaminergic, alpha(1) and alpha(2) noradrenergic receptors by a preliminary binding screen. The results show weak affinity to 5-HT(1A) for the aqueous extract. Both extracts showed affinity for D(1) receptors, but only for the methanolic extract the IC(50) value was determinable (30.14 microg/ml). No affinity for 5-HT(2A), 5-HT(2C) serotononergic receptors, alpha(1) and alpha(2) noradrenergic receptors and D(2) receptors was recorded for the extracts.

Endothelin-1 triggers placental oxidative stress pathways: putative role in preeclampsia.

CONTEXT: Preeclampsia (PE) is a disorder that occurs only during pregnancy. The placenta has a controlling role in this condition. Recent literature suggests that the oxidative stress is a component of PE and plays a main role in the link between decreased placental perfusion and the impaired function of maternal endothelium. OBJECTIVE: Because the human placenta expresses endothelin-1 (ET-1) and its circulating levels are high in pregnancies complicated with PE, the present study investigated the role of ET-1 on placental oxidative stress pathways. DESIGN: Human placental explants, JEG-3, and primary cytotrophoblast cells were cultured with increasing ET-1 concentrations for 6 and 24 h. SETTING: The study was conducted at tertiary clinical care centers in Siena and Padova, Italy. INTERVENTIONS: Human placental explants, JEG-3, and primary cytotrophoblast cells were used to test ET-1 effect. MAIN OUTCOME MEASURE(S): The main outcome measure was ET-1 mRNA and its receptor mRNAs, type A and B, detection by RT-PCR. The common markers of oxidative stress [malondialdehyde (MDA), glutathione (GSH), glutathione disulfide (GSSG), ascorbic acid (AA)] as well as cell proliferation and vitality were measured after stimulation periods. RESULTS: ET-1 inhibits cell proliferation and vitality and triggers oxidative stress in the human placenta by altering the balance between oxidant (increased MDA levels) and antioxidant (decreased GSH, GSSG, and AA) forces in favor of oxidation. CONCLUSIONS: Because MDA damages endothelial cells, whereas GSH, GSSG, and AA protect them, we postulate that ET-1 may be one of the key links between primary placental disorders and the systemic endothelial dysfunction of PE.

Unique Michael addition-initiated domino reaction for the stereoselective synthesis of functionalized macrolactones from alpha-nitroketones in water.

[reaction: see text]. A unique domino reaction of alpha-nitrocycloalkanones with alpha-alkyl alpha,beta-unsaturated aldehydes in aqueous base was discovered, leading to the one-pot synthesis of hitherto unknown functionalized, bridged, bicyclic lactones containing 10-, 11-, 13-, and 15-membered rings. The structures of these heterocyclic compounds, containing also an unusual 6-hydroxy-1,2-oxazine ring, were determined by spectral and single-crystal X-ray diffraction studies.

Immunotherapy of renal cell carcinoma with granulocyte macrophage colony stimulating factor and very low dose interleukin-2.

We have performed a translational phase II trial testing an original immunotherapy schedule based on the monthly subcutaneous (s.c.) administration of hrGM-CSF (days 1 through 5) and very low dose hrIL-2 (days 6 through 15) in 19 patients with metastatic renal cell carcinoma. Bone pain, first dose reaction to GM-CSF, asthenia and fever were the most common side effects. A partial response, and a disease stabilization were respectively observed in 4 and 11 cases, with a rate of objective response and a disease control rate respectively of 21% and 79%. We recorded a time to progression of 9 months and a 2- and 3-year survival respectively of 42% (8/19 patients) and 26% (5/19 patients). Our results suggest that this GM-CSF/hrIL-2 combination is active and well tolerated in patients with renal cell carcinoma and deserves to be investigated in larger comparative trials.

Immunosuppression and oxidative stress induced by acute and chronic exposure to cocaine in rat.

The objective of the present study was to verify if immunosuppression caused by cocaine (CO) can be mediated, at least in part, by increased formation of oxidative metabolites and reactive oxygen species (ROS) in rat. Pharmacokinetics of cocaine and its metabolites, cell-mediated immune function and cytokines production, biomarkers of cell redox state maintenance and lipidic peroxidation, and variations of activity in the enzymatic systems involved in cell antioxidant defence were measured in spleen of Wistar rats acutely and chronically treated with cocaine.C(max), AUC, and t(1/2) of norcocaine (NC) significantly increased after chronic exposure to cocaine while kinetic parameters of benzoylecgonine (BE) significantly decreased. A decrease in cultured T-lymphocytes proliferation and natural killer (NK) cell activity, a high increase of immunosuppressive cytokines and a switch from Th1-type cytokines to Th2-type cytokines together with an unbalance toward anti-inflammatory cytokines recovered within 4 h after acute treatment while subsisted for 14 days after chronic treatment. A significant increase in ascorbic acid (AA), reduced glutathione and glutathione reductase (GR) with a simultaneous decrease in oxidized glutathione were observed in the first hours after acute administration. Conversely, the increase in oxidized glutathione and malondialdehyde (MDA) production and the simultaneous depletion of reduced glutathione and ascorbic acid persisted at least 24 h after chronic cocaine treatment as well as the increase in the activities of glutathione reductase, glutathione peroxidase (GPx) and superoxide dismutase (SOD). The results suggest that chronic cocaine administration affects cellular enzyme and non-enzyme-mediated antioxidant defence systems and promotes immunotoxicity in rat. Cocaine N-oxidative metabolism may be an indirect contributor, via oxidative stress.

Impaired uterine artery blood flow at mid gestation and low levels of maternal plasma corticotropin-releasing factor.

OBJECTIVE: Corticotropin-releasing factor (CRF) is a placental neuropeptide that plays a role in the control of uteroplacental blood flow regulation. Because CRF has a relaxant effect on uterine vasculature in pregnant rats, we aimed to evaluate mid-gestation plasma CRF levels in women with impaired uterine artery blood flow. METHODS: Maternal plasma CRF was assayed by specific radioimmunoassay, and uterine artery resistance index (RI) was assessed by Doppler evaluation at 22-24 weeks' gestation in 55 healthy pregnant women, of whom 24 showed a unilateral or bilateral uterine artery notch, reflecting resistance. Statistical analysis was performed by the Kruskal-Wallis test followed by the post hoc Dunn's test and the Spearman rank test. RESULTS: The mean uterine artery RI was significantly (P <.001) higher in women with a notch than in healthy controls. Mean +/- standard error of the mean maternal plasma CRF levels were significantly lower in women with a unilateral (168.45 +/- 27.5 pg/mL; P <.01) or bilateral (186.07 +/- 34.5 pg/mL; P <.001) uterine artery notch than in healthy control pregnant women (375.06 +/- 21.77 pg/mL). Although no difference was found in CRF levels between patients with a unilateral or bilateral uterine artery notch, a significant inverse correlation was found between the mean RI and maternal plasma CRF levels (Spearman r = -0.6540; 95% confidence interval, -0.7865, -0.4640; P <.001). CONCLUSION: Reduced levels of circulating CRF were associated with increased uterine artery resistance, which supports the hypothesis that CRF may regulate uterine artery tone at mid gestation.

Pharmacokinetics of intraarterial mitomycin C in hypoxic hepatic infusion with embolization in the treatment of liver metastases.

1. The pharmacokinetics of mitomycin C (MMC) was evaluated during hypoxic hepatic infusion (HHMI) with arterial embolization for the treatment of unresectable liver metastases. 2. Ten patients with hepatic metastases from colorectal cancer were considered. Antiblastic infusion with MMC (20 mg/m2 at 30 ml/min) was initiated after 10 min of hepatic arterial occlusion. Peripheral venous blood samples were collected at different time intervals. MMC was assayed by high-pressure liquid chromatography (HPLC), and pharmacokinetic parameters were determined using an open, two-compartment model and linear kinetics. 3. Cmax of MMC during HHMI was 708 +/- 336.6 ng/ml, and tmax was 9.3 +/- 1.1 min. The plasma concentration-time curve showed a t1/2 alpha ranging from 1.5 to 9 min, followed by a t1/2 beta ranging from 31 to 93 min. The Cltot was 35.5 l/h/m2 with an area under the plasma concentration-time curve (AUC) ranging from 251 to 850 micrograms h/l. The same AUC parameter standardized for the amount of MMC was 15.5 mg-1. The HHMI model that we used revealed a significant increase in Cltot and a reduction in AUC when compared to the locoregional intraarterial and peripheral intravenous models (p < .001). 4. The reduction in AUC following HHMI explains the limited systemic toxicity in treated patients, with a greater total tumor exposure to the drug and improved drug activation.

Domperidone and long QT syndrome.

Domperidone is a prokinetic agent widely prescribed in adults and children with gastrointestinal disorders. Recently several Regulatory Agencies published safety information on the risk of long QT syndrome associated with the use of domperidone. We conducted a literature review using PubMed (1966 - Jan 2010) with the aim to locate articles on ventricular arrhytmias, Sudden Cardiac Death (SCD), long QT syndrome and Torsade de Pointes (TdP) following domperidone administration. Twenty-two papers were included in the review: three studies in vitro models, one animal study, five case reports/series, twelve clinical studies and one editorial. In vitro studies demonstrated cardiac electrophysiological effects of domperidone on the rapid component of the cardiac delayed rectifier K(+) current (I(Kr)) through the blockade of channels encoded by the Human Ether-a-go-go Related Gene (HERG). A total of fourteen cases of cardiotoxicity following intravenous (12 cases) or oral (2 cases) domperidone administration from case reports/series were identified. A case - control study, performed on a general practice observational database reported an OR: 3.8 (95% CI: 1.5-9.7) for SCD after domperidone exposure. Prescribers and other healthcare professionals should take into account the risk of QT syndrome in domperidone users and avoid administering domperidone in patients concomitantly taking strong CYP3A4 inhibitors or other QT prolonging drugs.