Drug development in Parkinson's disease: From emerging molecules to innovative drug delivery systems

Current treatments for Parkinson’s disease (PD) are aimed at addressing motor symptoms but there is no therapy focused on modifying the course of the disease. Successful treatment strategies have been so far limited and brain drug delivery remains a major challenge that restricts its treatment. This review provides an overview of the most promising emerging agents in the field of PD drug discovery, discussing improvements that have been made in brain drug delivery for PD. It will be shown that new approaches able to extend the length of the treatment, to release the drug in a continuous manner or to cross the blood brain barrier and target a specific region are still needed. Overall, the results reviewed here show that there is an urgent need to develop both symptomatic and disease-modifying treatments, giving priority to neuroprotective treatments. Promising perspectives are being provided in this field by rasagiline and by neurotrophic factors like glial cell line-derived neurotrophic factor. The identification of disease-relevant genes has also encouraged the search for disease-modifying therapies that function by identifying molecularly-targeted drugs. The advent of new molecular and cellular targets like α-synuclein, leucine-rich repeat serine/threonine protein kinase 2 or parkin, among others, will require innovative delivery therapies. In this regard, drug delivery systems (DDS) have shown great potential for improving the efficacy of conventional and new PD therapy and reducing its side effects. The new DDS discussed here, which include microparticles, nanoparticles and hydrogels among others, will probably open up possibilities that extend beyond symptomatic relief. However, further work needs to be done before DDS become a therapeutic option for PD patients

Improvement of nutritional properties of Chorizo de Pamplona by replacement of pork backfat with soy oil.

Dry fermented sausages with a partial substitution of 15, 20 and 25% of pork backfat by pre-emulsified soy oil were prepared. No differences were detected in the water, protein and fat content between control and modified sausages. Cholesterol amount scarcely decreased in the modified sausages (92.96 mg/100 g product in control sausages, 87.71 mg/100 g in sausages prepared with 25% of substitution). No increase in oxidation was detected through chemical or sensory analysis in modified sausages. Saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA) in control products were 37.83 and 45.78 g/100 g of total fatty acid, respectively, decreasing in the modified formulations, to 32.81 and 42.09 g/100 g of total fatty acid in the 25% replacement products, respectively. Polyunsaturated fatty acids (PUFA) increased from 15.22 (control) to 23.96 g/100 g of total fatty acid (25% product) due to the significant increase in linoleic and α-linolenic acids when soy oil was added. In relation to texture profile analysis (TPA), hardness and springiness did not show significant differences among products. The instrumental measured colours were comparable with that of commercial products. Sensory evaluation of most of the modified sausages did not show significant differences with regard to the control

Functional dry fermented sausages manufactured with high levels of n-3 fatty acids: nutritional benefits and evaluation of oxidation

Dry fermented sausages including fish oil extracts rich in n-3 fatty acids were manufactured in order to obtain functional products, and their nutritional advantages and effects on oxidation process were compared with traditional ones. Modified products were manufactured with 5.3 g kg−1 and 10.7 g kg−1 of fish oil extract. Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) contents per 100 g of dry fermented sausages were 0.15 g and 0.13 g for the lower rate of addition and 0.33 g and 0.26 g, respectively, for the higher rate, compared with only 0.01 g EPA and 0.03 g DHA in the control products. No significant differences were found in the other fatty acids. A decrease in the n-6/n-3 ratios from 16.14 in the control to 7.78 in batch A and 5.32 in batch B was achieved. Thiobarbituric acid reactive substances values were similar in control (0.31 ppm) and in batch A products (0.34 ppm) but increased significantly in batch B products (1.22 ppm). No statistical differences were observed among batches for the content of cholesterol oxidation products (2.36–2.43 µg g−1 fat) leading to similar percentages of oxidation. 7-Ketocholesterol, considered an indicator of oxidation, was not present in any sample. Values obtained for L* and hue (arctg b*/a*) were comparable with those of meat products. Although no effect was observed in cholesterol oxidation product formation and instrumental measurements of colour, the highest level of n-3 fatty acid seemed to accelerate the oxidation process significantly. Copyright © 2004 Society of Chemical Industr

AGAP2: modulating TGFβ1-signalling in the regulation of liver fibrosis

AGAP2 (Arf GAP with GTP-binding protein-like domain, Ankyrin repeat and PH domain 2) isoform 2 is a protein that belongs to the Arf GAP (GTPase activating protein) protein family. These proteins act as GTPase switches for Arfs, which are Ras superfamily members, being therefore involved in signaling regulation. Arf GAP proteins have been shown to participate in several cellular functions including membrane trafficking and actin cytoskeleton remodeling. AGAP2 is a multi-tasking Arf GAP that also presents GTPase activity and is involved in several signaling pathways related with apoptosis, cell survival, migration, and receptor trafficking. The increase of AGAP2 levels is associated with pathologies as cancer and fibrosis. Transforming growth factor beta-1 (TGF-β1) is the most potent pro-fibrotic cytokine identified to date, currently accepted as the principal mediator of the fibrotic response in liver, lung, and kidney. Recent literature has described that the expression of AGAP2 modulates some of the pro-fibrotic effects described for TGF-β1 in the liver. The present review is focused on the interrelated molecular effects between AGAP2 and TGFβ1 expression, presenting AGAP2 as a new player in the signaling of this pro-fibrotic cytokine, thereby contributing to the progression of hepatic fibrosis

Effective GDNF brain delivery using microspheres-A promising strategy for Parkinson's disease

Glial cell line-derived neurotrophic factor (GDNF) has shown promise in the treatment of neurodegenerative disorders of basal ganglia origin such us Parkinson\u27s disease (PD). In this study, we investigated the neurorestorative effect of controlled GDNF delivery using biodegradable microspheres in an animal model with partial dopaminergic lesion. Microspheres were loaded with N-glycosylated recombinant GDNF and prepared using the Total Recirculation One-Machine System (TROMS). GDNF-loaded microparticles were unilaterally injected into the rat striatum by stereotaxic surgery two weeks after a unilateral partial 6-OHDA nigrostriatal lesion. Animals were tested for amphetamine-induced rotational asymmetry at different times and were sacrificed two months after microsphere implantation for immunohistochemical analysis. The putative presence of serum IgG antibodies against rat glycosylated GDNF was analyzed for addressing safety issues. The results demonstrated that GDNF-loaded microspheres, improved the rotational behavior induced by amphetamine of the GDNF-treated animals together with an increase in the density of TH positive fibers at the striatal level. The developed GDNF-loaded microparticles proved to be suitable to release biologically active GDNF over up to 5 weeks in vivo. Furthermore, none of the animals developed antibodies against GDNF demonstrating the safety of glycosylated GDNF use

New formulations for healthier dry fermented sausages: a review.

An excessive intake of meat products, particularly dryfermented sausages, is not recommended from a health point of view, at least for some population groups, due to their high level of sodium and animal fat. Many efforts of the meat industry are focused on the development of new products with better nutritional properties than traditional ones. KCl, CaCl2, and/or calcium ascorbate, among others, have been assessed as partial substitutes of NaCl, giving products with acceptable sensory quality, smaller amounts of sodium and being sometimes a significant source of potassium or calcium. In relation to fat, recent research has focused on the use of different types of fibres and vegetable oils as partial substitutes of pork backfat. The use of fibres results in low-fat and low-energy products. The use of vegetable oils results in products with healthier fatty acid profiles. Aspects related to improving sensory properties and control of oxidation should be taken into account in future research

Effect of fat level and partial replacement of pork backfat with olive oil on processing and quality characteristics of fermented sausages

Six formulations of dry fermented sausages were produced in three replications with three initial fat levels (30, 20 and 10%) and two levels (0 and 20%) of pork backfat replacing olive oil. After 4 weeks of fermentation and ripening the fat content of the treatments with 30, 20 and 10% fat level ranged from 38.86 to 43.60%, 25.56 to 26.86% and 19.01 to 20.14%, respectively. Fat level affected (P<0.05) the weight losses, the chemical composition, the Gram -ve bacterial count, the lightness, the texture and the appearance of fermented sausages. Replacing 20% of pork backfat by olive oil affected (P<0.05) the lightness and yellowness of sausages. Fat-reduced sausages without olive oil and low-fat sausages with olive oil had the highest score for odour and taste. However, the appearance of fat-reduced sausages was just acceptable while that of low-fat sausages was unacceptable, because the surface was intensively wrinkled and case hardening had developed. Further research is needed to improve the appearance of these sausages

Terapias neuroprotectoras y neurorestauradoras en el tratamiento de la enfermedad de Parkinson

Parkinson's disease is the second most common neurodegenerative disorder after Alzheimer's disease. Current therapies are symptomatic and, although these therapies are efficacious during the early stages of the disease, they present important side effects when they are used for a long time. The ideal therapy would be the one that would slow down or stop the progression of the disease. This can be achieved, for instance, with neuroprotective and neurorestorative therapies. Among them, cell therapy and therapy with trophic factors such as glial cell line derived neurotrophic factor (GDNF) are the most challenging and promising ones for the scientific community. Although the use of GDNF as a treatment for Parkinson s disease was proposed several years ago, it is necessary to develop alternative strategies to deliver GDNF appropriately to concrete areas of the brain. Here, the use of microspheres as the most suitable tool for the administration of this neurotrophic factor is discussed

Long-term neuroprotection and neurorestoration by glial cell-derived neurotrophic factor microspheres for the treatment of parkinson's disease

BACKGROUND: Glial cell-derived neurotrophic factor is a survival factor for dopaminergic neurons and a promising candidate for the treatment of Parkinson's disease. However, the delivery issue of the protein to the brain still remains unsolved. Our aim was to investigate the effect of long-term delivery of encapsulated glial cell-derived neurotrophic factor within microspheres. METHODS: A single dose of microspheres containing 2.5 μg of glial cell-derived neurotrophic factor was implanted intrastriatally in animals 2 weeks after a 6-hydroxydopamine lesion. RESULTS: The amphetamine test showed a complete behavioral recovery after 16 weeks of treatment, which was maintained until the end of the study (week 30). This effect was accompanied by an increase in dopaminergic striatal terminals and neuroprotection of dopaminergic neurons. CONCLUSIONS: The main achievement was the long-term neurorestoration in parkinsonian animals induced by encapsulated glial cell-derived neurotrophic factor, suggesting that microspheres may be considered as a means to deliver glial cell-derived neurotrophic factor for Parkinson's disease treatment. © 2011 Movement Disorder Society

Production of highly pure human glycosylated GDNF in a mammalian cell line

The administration of glial cell line-derived neurotrophic factor (GDNF) has emerged as a promising strategy for the treatment of several diseases of the nervous system as Parkinson's disease, amyotrophic lateral sclerosis, spinal cord injury and nerve regeneration as well as ocular diseases and drug addictions. A procedure for the purification of human recombinant glycosylated GDNF using a mammalian expression system as the source of the protein is discussed in the present paper. The neurotrophic factor was purified using cation exchange chromatography and gel filtration. A human cell line was chosen as the source of therapeutic protein, since a recombinant protein with a structure and glycosylation pattern equivalent to the native form is desirable for its prospective therapeutic utilization. The activity of the highly pure protein obtained was confirmed with a cell-based bioassay. The purified protein is suitable for its in vivo evaluation in animals and for possible subsequent clinical application