Novel Curcumin-Diethyl Fumarate Hybrid as a Dualistic GSK-3ß Inhibitor/Nrf2 Inducer for the Treatment of Parkinson's Disease.

Common copathogenic factors, including oxidative stress and neuroinflammation, are found to play a vital role in the development of neurodegenerative disorders, including Alzheimer's disease (AD) and Parkinson's disease (PD). Nowadays, owing to the multifactorial character of the diseases, no effective therapies are available, thus underlying the need for new strategies. Overexpression of the enzyme GSK-3ß and downregulation of the Nrf2/ARE pathway are responsible for a decrease in antioxidant defense effects. These pieces of evidence underline the usefulness of dual GSK-3ß inhibitors/Nrf2 inducers. In this regard, to design a dual modulator, the structures of a curcumin-based analogue, as GSK-3ß inhibitor, and a diethyl fumarate fragment, as Nrf2 inducer, were combined. Among the hybrids, 5 and 6 proved to effectively inhibit GSK-3ß, while 4 and 5 showed a marked ability to activate Nrf2 together to increase the neuronal resistance to oxidative stress. These last pieces of evidence translated into specific neuroprotective effects of 4 and 5 against PD pathological events including neurotoxicity elicited by α-synuclein aggregates and 6-hydroxydopamine. Hybrid 5 also showed neuroprotective effects in a C. elegans model of PD where the activation of GSK-3ß is intimately involved in Nrf2 regulation. In summary, 5 emerged as an interesting multitarget derivative, valuable to be exploited in a multitarget PD perspective.

Tau-Centric Multitarget Approach for Alzheimer's Disease: Development of First-in-Class Dual Glycogen Synthase Kinase 3ß and Tau-Aggregation Inhibitors.

Several findings propose the altered tau protein network as an important target for Alzheimer's disease (AD). Particularly, two points of pharmacological intervention can be envisaged: inhibition of phosphorylating tau kinase GSK-3ß and tau aggregation process. On the basis of this consideration and on our interest in multitarget paradigms in AD, we report on the discovery of 2,4-thiazolidinedione derivatives endowed with such a profile. 28 and 30 displayed micromolar IC50 values toward GSK-3ß, together with the capacity of inhibiting AcPHF6 aggregation of 60% and 80% at 10 µM, respectively. In addition, they showed PAMPA-BBB permeability, together with a suitable cellular safety profile. 30 also displayed inhibition of both K18 and full-length tau aggregations. Finally, both compounds were able to improve cell viability in an okadaic acid-induced neurodegeneration cell model. To the best of our knowledge, 28 and 30 are the first balanced, nontoxic, dual-acting compounds hitting tau cascade at two different hubs.

Identification of new allosteric sites and modulators of AChE through computational and experimental tools.

Allosteric sites on proteins are targeted for designing more selective inhibitors of enzyme activity and to discover new functions. Acetylcholinesterase (AChE), which is most widely known for the hydrolysis of the neurotransmitter acetylcholine, has a peripheral allosteric subsite responsible for amyloidosis in Alzheimer's disease through interaction with amyloid ß-peptide. However, AChE plays other non-hydrolytic functions. Here, we identify and characterise using computational tools two new allosteric sites in AChE, which have allowed us to identify allosteric inhibitors by virtual screening guided by structure-based and fragment hotspot strategies. The identified compounds were also screened for in vitro inhibition of AChE and three were observed to be active. Further experimental (kinetic) and computational (molecular dynamics) studies have been performed to verify the allosteric activity. These new compounds may be valuable pharmacological tools in the study of non-cholinergic functions of AChE.

Subtly Modulating Glycogen Synthase Kinase 3 ß: Allosteric Inhibitor Development and Their Potential for the Treatment of Chronic Diseases.

Glycogen synthase kinase 3 ß (GSK-3ß) is a central target in several unmet diseases. To increase the specificity of GSK-3ß inhibitors in chronic treatments, we developed small molecules allowing subtle modulation of GSK-3ß activity. Design synthesis, structure-activity relationships, and binding mode of quinoline-3-carbohydrazide derivatives as allosteric modulators of GSK-3ß are presented here. Furthermore, we show how allosteric binders may overcome the ß-catenin side effects associated with strong GSK-3ß inhibition. The therapeutic potential of some of these modulators has been tested in human samples from patients with congenital myotonic dystrophy type 1 (CDM1) and spinal muscular atrophy (SMA) patients. We found that compound 53 improves delayed myogenesis in CDM1 myoblasts, while compounds 1 and 53 have neuroprotective properties in SMA-derived cells. These findings suggest that the allosteric modulators of GSK-3ß may be used for future development of drugs for DM1, SMA, and other chronic diseases where GSK-3ß inhibition exhibits therapeutic effects.

Application of BACE1 immobilized enzyme reactor for the characterization of multifunctional alkaloids from Corydalis cava (Fumariaceae) as Alzheimer's disease targets.

In our ongoing study focused on Corydalis cava (Fumariaceae), used in folk medicine in the treatment of memory dysfunctions, we have investigated fifteen previously isolated alkaloids for their potential multifunctional activity on Alzheimer's disease (AD) targets. Determination of ß-site amyloid precursor protein cleaving enzyme 1 (BACE1) inhibition was carried out using a BACE1-Immobilized Enzyme Reactor (IMER) by validating the assay with a multi-well plate format Fluorescence Resonance Energy Transfer (FRET) assay. Seven alkaloids out of fifteen were found to be active, with (-)-corycavamine (3) and (+)-corynoline (5) demonstrating the highest BACE1 inhibition activity, in the micromolar range, in a concentration dependent manner. BACE1-IMER was found to be a valid device for the fast screening of inhibitors and the determination of their potency. In a permeation assay (PAMPA) for the prediction of blood-brain barrier (BBB) penetration, the most active compounds, (-)-corycavamine (3) and (+)-corynoline (5), were found to be able to cross the BBB. Not all compounds showed activity against glycogen synthase kinase-3ß (GSK-3ß) and casein kinase-1δ (CK-1δ). On the basis of the reported results, we found that some C. cava alkaloids have multifunctional activity against AD targets (prolyl oligopeptidase, cholinesterases and BACE1). Moreover, we tried to elucidate the treatment effectivity (rational use) of its extract in memory dysfunction in folk medicine.

Supercritical fluid extraction of grape seeds: extract chemical composition, antioxidant activity and inhibition of nitrite production in LPS-stimulated Raw 264.7 cells.

Grape by-products are a rich source of bioactive compounds having broad medicinal properties, but are usually wasted from juice/wine processing industries. The present study investigates the use of supercritical fluid extraction (SFE) for obtaining an extract rich in bioactive compounds. First, some variables involved in the extraction were applied. SFE conditions were selected based on the oil mass yield, fatty acid profile and total phenolic composition. As a result, 40 °C and 300 bar were selected as operational conditions. The phenolic composition of the grape seed oil was determined using LC-DAD. The antioxidant activity was determined by ABTS and DPPH assays. For the anti-inflammatory activity the inhibition of nitrite production was assessed. The grape seed oil extracted was rich in phenolic compounds and fatty acids with significant antioxidant and anti-inflammatory activities. From these results, added economic value to this agroindustrial residue is proposed using environmentally friendly techniques.

Effect of dietary supplementation with red wine extract or vitamin E, in combination with linseed and fish oil, on lamb meat quality.

Thirty lambs were assigned to the following treatments: control diet (C) rich in omega-3 fatty acids; C plus 900ppm red wine extract (RWE), or C plus 300ppm vitamin E (VE). Oxidative stability and sensory properties of chops stored in MAP (70% O2/30% CO2) during 12days were evaluated. Chops from the VE group showed lower lipid oxidation (p<0.001) and protein carbonylation (p<0.05), stable omega-3 fatty acids proportions and overall liking sensory scores (p<0.05). Dietary RWE supplementation did not influence oxidative stability of chops, however levels of C20:5n-3 were greater (p<0.05) and n-6/n-3 ratio (p<0.01) was lower, relative to controls.

Effect of dietary supplementation with either red wine extract or vitamin E on the volatile profile of lamb meat fed with omega-3 sources.

The effect of dietary supplementation with either vitamin E (300 ppm) or a red wine extract rich in polyphenols (900 ppm) in an omega-3 enriched concentrate on the volatile fraction of lamb meat was assessed. The effect of refrigerated storage (0 and 6 days) under high-oxygen atmospheres (70% O(2)/30% CO(2)) was also studied. Extraction and analysis of the volatile compounds was carried out by headspace solid-phase microextraction (HS-SPME) and GC-MS, respectively. Vitamin E supplementation led to lower levels of lipo-oxidation compounds, such as 2-heptanone and 1-penten-3-ol. The red wine extract was less efficient against lipid oxidation than vitamin E but more efficient than the control (no added antioxidants). The levels of numerous lipid-derived compounds were found to be lower after 6 days of storage which could be due to further interactions with protein-related compounds.

An economic evaluation of pregabalin versus usual care in the management of community-treated patients with refractory painful diabetic peripheral neuropathy in primary care settings.

OBJECTIVE: To estimate the cost-effectiveness of pregabalin versus usual care (UC) in the management of community-treated patients with refractory painful diabetic peripheral neuropathy (pDPN) in primary care settings (PCS) in Spain. METHODS: Data was extracted from a 12-week registry study assessing costs of neuropathic pain in Spain. Pregabalin-naïve outpatients treated with UC or newly prescribed pregabalin were selected for inclusion in the cost-effectiveness analysis. Effectiveness was expressed as quality-adjusted life years (QALY) gain. Perspectives of the Spanish National Health System (NHS) and society (2006) were applied for cost calculations. Results were expressed as incremental cost-effectiveness ratio (ICER). Bootstrapping techniques (10,000 re-samples) were used to obtain the probabilistic ICER and the cost-effectiveness acceptability curve. RESULTS: A total of 189 patients were included in the economic analysis. Compared with UC, pregabalin was associated with higher QALY gain in a period of 12-weeks; 0.0406±0.0343 versus 0.0285±0.0350 (p=0.167). Overall total costs (€1368±1229 vs. €1258±1474; p=0.587) and healthcare costs (€628±590 vs. €469±420; p=0.134) were similar for both pregabalin and UC, respectively. ICERs for pregabalin varied from €5302 (95% CI: dominant; €144,105) for total costs to €14,381 (dominant; €115,648) for healthcare costs. Probabilistic sensitivity analyses showed that 79-84% of ICERs were below the threshold of €30,000/QALY. CONCLUSION: This study suggests that pregabalin may be cost-effective in the management of community-treated refractory outpatients, with pDPN when compared with usual care in the primary care setting in Spain. These findings may help policy makers when making health decision in the management of diabetes in the community.

Pain alleviation and patient-reported health outcomes following switching to pregabalin in individuals with gabapentin-refractory neuropathic pain in routine medical practice.

BACKGROUND: It has been suggested that peripheral neuropathic pain (PNP) may affect up to 3% of the general population. PNP has a substantial negative impact on patient functioning and quality of life, including reduced productivity and increased consumption of healthcare resources. OBJECTIVE: The objective of this study was to analyse changes in pain and patient-reported health outcomes following switching to pregabalin treatment in patients with gabapentin-refractory PNP as part of routine clinical practice in Spanish primary-care settings. METHODS: This was an open-label, non-randomized, post hoc analysis of a 12-week, multicentre, non-interventional cost-of-illness study. The study involved primary-care physicians and was carried out between September 2005 and April 2006. Patients were aged at least 18 years and had chronic, treatment-refractory PNP. The analysis included all pregabalin-naïve patient switches that had previously shown an inadequate response to gabapentin. The following variables were assessed before and after pregabalin therapy: pain (Short-Form McGill Pain Questionnaire [SF-MPQ]), disability level (Sheehan Disability Scale [SDS]), symptoms of anxiety and depression (Hospital Anxiety and Depression Scale [HADS]), self-perceived sleep quality and quantity (Medical Outcomes Study [MOS] Sleep Scale), and health-related quality of life (EuroQol [EQ] five-dimension [5D] questionnaire). RESULTS: A total of 174 patients with an inadequate response to gabapentin switched to pregabalin at the beginning of the study. After 12 weeks of pregabalin therapy, alone or in combination with other analgesic drugs, significant and clinically relevant improvements were observed in pain severity [mean (SD) SF-MPQ change: -31.9 (22.1) points; 42.5% being responders (pain reduction ≥50%)], disability [SDS score: -7.5 (6.2) vs baseline], affective symptoms [HADS depression: -4.5 (3.9); HADS anxiety: -4.1 (3.7)], sleep [MOS summary index: -17.2 (16.6)], and health status [EQ-5D visual analogue scale (VAS): 27.3 (18.2)], with an overall gain of 0.040 (0.031) quality-adjusted life-years. CONCLUSION: These findings suggest that pregabalin could be a valid treatment alternative for the management of patients with gabapentin-refractory peripheral neuropathic pain in primary-care settings under real-life conditions of care. Our data show that patients who were switched to pregabalin, either as monotherapy or in combination with other analgesics, showed substantial and clinically relevant improvements in relieving pain and related symptoms.

New tacrine-4-oxo-4H-chromene hybrids as multifunctional agents for the treatment of Alzheimer's disease, with cholinergic, antioxidant, and ß-amyloid-reducing properties.

By using fragments endowed with interesting and complementary properties for the treatment of Alzheimer's disease (AD), a new family of tacrine-4-oxo-4H-chromene hybrids has been designed, synthesized, and evaluated biologically. The tacrine fragment was selected for its inhibition of cholinesterases, and the flavonoid scaffold derived from 4-oxo-4H -chromene was chosen for its radical capture and ß-secretase 1 (BACE-1) inhibitory activities. At nano- and picomolar concentrations, the new tacrine-4-oxo-4H-chromene hybrids inhibit human acetyl- and butyrylcholinesterase (h-AChE and h-BuChE), being more potent than the parent inhibitor, tacrine. They are also potent inhibitors of human BACE-1, better than the parent flavonoid, apigenin. They show interesting antioxidant properties and could be able to penetrate into the CNS according to the in vitro PAMPA-BBB assay. Among the hybrids investigated, 6-hydroxy-4-oxo- N-{10-[(1,2,3,4-tetrahydroacridin-9-yl)amino]decyl}-4 H-chromene-2-carboxamide (19) shows potent combined inhibition of human BACE-1 and ChEs, as well as good antioxidant and CNS-permeable properties.

Phosphodiesterase 7 inhibition preserves dopaminergic neurons in cellular and rodent models of Parkinson disease.

BACKGROUND: Phosphodiesterase 7 plays a major role in down-regulation of protein kinase A activity by hydrolyzing cAMP in many cell types. This cyclic nucleotide plays a key role in signal transduction in a wide variety of cellular responses. In the brain, cAMP has been implicated in learning, memory processes and other brain functions. METHODOLOGY/PRINCIPAL FINDINGS: Here we show a novel function of phosphodiesterase 7 inhibition on nigrostriatal dopaminergic neuronal death. We found that S14, a heterocyclic small molecule inhibitor of phosphodiesterase 7, conferred significant neuronal protection against different insults both in the human dopaminergic cell line SH-SY5Y and in primary rat mesencephalic cultures. S14 treatment also reduced microglial activation, protected dopaminergic neurons and improved motor function in the lipopolysaccharide rat model of Parkinson disease. Finally, S14 neuroprotective effects were reversed by blocking the cAMP signaling pathways that operate through cAMP-dependent protein kinase A. CONCLUSIONS/SIGNIFICANCE: Our findings demonstrate that phosphodiesterase 7 inhibition can protect dopaminergic neurons against different insults, and they provide support for the therapeutic potential of phosphodiesterase 7 inhibitors in the treatment of neurodegenerative disorders, particularly Parkinson disease.

Manejo de la enfermedad diarreica en atención primaria de salud / Managemen of acute diarrheic disease in primary health care

Se realizó un estudio descriptivo en los consultorios urbanos del policlínico área sur de Sancti Spíritus en el período comprendido de enero de 1997 a diciembre del 2000 con el objetivo de determinar el manejo de la enfermedad diarreica aguda en la atención primaria de salud. Se obtuvo como resultados que los medicamentos no se utilizan en el año 2000 en el 66,32 por ciento de los niños con diarreas. La terapia de rehidratación oral se utilizó en el 64,289 por ciento de los niños en el año 2000. Se arribó a las siguientes conclusiones que las sales de hidratación oral se utilizaron en todos los pacientes estudiados, los remedios caseros se fueron utilizando menos en el transcurso de los años y en el seguimiento y control de los mismos mejora considerablemente. No obstante se considera que se puede mejorar aún más en el manejo de está enfermedad(AU)

A descriptive study was made in the urban clinics of the South Polyclinic area of Sancti Spíritus in the period from January of 1997 to December of 2000 with the objective of determining the management of acute diarrheic disease in primary health care. It was obtained as results that medications are not used in the year 2000 in 66,32 percent of the children with diarrheas. Oral rehydration therapy was used in 64,289 percent of the children in the year 2000. The following conclusions were drawn: oral rehydration salts were used in all the patients studied, the use of homemade remedies decreased with the passing of years and their follow-up and control improved considerably. Nevertheless, it is considered that management of this disease can be further improved(AU)

Manejo de la enfermedad diarreica en atención primaria de salud / Managemen of acute diarrheic disease in primary health care

Se realizó un estudio descriptivo en los consultorios urbanos del policlínico área sur de Sancti Spíritus en el período comprendido de enero de 1997 a diciembre del 2000 con el objetivo de determinar el manejo de la enfermedad diarreica aguda en la atención primaria de salud. Se obtuvo como resultados que los medicamentos no se utilizan en el año 2000 en el 66,32% de los niños con diarreas. La terapia de rehidratación oral se utilizó en el 64,289% de los niños en el año 2000. Se arribó a las siguientes conclusiones que las sales de hidratación oral se utilizaron en todos los pacientes estudiados, los remedios caseros se fueron utilizando menos en el transcurso de los años y en el seguimiento y control de los mismos mejora considerablemente. No obstante se considera que se puede mejorar aún más en el manejo de está enfermedad.]AU[