High-throughput dissolution/permeation screening : a 96-well two-compartment microplate approach

Early formulation screening can alleviate development of advanced oral drug formulations, such as amorphous solid dispersions (ASDs). Traditionally, dissolution is used to predict ASD performance. Here, a high-throughput approach is described that simultaneously screens drug dissolution and permeation employing a two-compartment 96-well plate. Freeze-drying from hydro-alcoholic solutions was used to prepare amorphous formulations. The screening approach was tested on amorphous and crystalline tadalafil formulations with and without Soluplus®. The workflow consisted of: 1) dispersion of the formulations; 2) incubation within the two-compartment plate, where a dialysis membrane separated donor (dispersed formulation) and acceptor; 3) sampling (donor and acceptor), where donor samples were centrifuged to remove non-dissolved material; and 4) quantification by UHPLC-UV. To identify optimal screening conditions, the following parameters were varied: dispersion medium (buffer / biomimetic media), acceptor medium (buffer / surfactant solutions), and incubation time (1, 3, and 6 h). Surfactants (acceptor) increased tadalafil permeation. Biomimetic medium (donor) enhanced dissolution, but not permeation, except for freeze-dried tadalafil, for which the permeated amount increased. The predictiveness was evaluated by comparing dissolution-/permeation-results with in vivo bioavailability. In general, both dissolution and permeation reflected bioavailability, whereof the latter was a better predictor. High-throughput dissolution/permeation is regarded promising for formulation screening

A dynamic in vitro permeation study on solid mono- and diacyl-phospholipid dispersions of celecoxib

The current study documents enhanced apparent solubility of the BCS class II drug celecoxib (CXB) when formulated as solid phospholipid dispersion (SPD) with either mono- or diacyl-phospholipids by freeze drying from hydro-alcoholic solvent. The enhanced solubility upon dispersion in buffer or fasted state simulated intestinal fluid (FaSSIF) is interpreted to be due to two effects: (1) amorphization of CXB, inducing supersaturation, which is also observed when CXB is freeze dried in the absence of phospholipids and (2) association of CXB with spontaneously forming colloidal structures, such as vesicles and/or micelles, promoting solubilization. The latter effect depended on the CXB-to-phospholipid ratio, where monoacyl-phospholipid was a more efficient solubilizer than diacyl-phospholipid. In the case of diacyl-phospholipid, solubilization also depended strongly on the dispersion medium, where FaSSIF induced a more pronounced solubilization effect than buffer. In contrast, a significantly enhanced in-vitro permeability of CXB across a biomimetic barrier (Permeapad®) was found only with low lipid contents up to a CXB to phospholipid mass-ratio of 1:10 or in the absence of phospholipid; above this critical ratio, permeability was not enhanced, i.e. comparable to that observed with a suspension of non-processed (crystalline) drug. This non-linear dissolution-/permeation-behavior was observed independently of (1) the type of phospholipid (monoacyl- or diacyl-) employed and (2) the dispersion medium (buffer or FaSSIF), despite the fact that different patterns of co-existing colloidal states were observed from mono-/diacyl-phospholipid formulations in buffer/FaSSIF (small bile salt micelles, intermediate size mixed micelles and large vesicular structures), assessed by asymmetric flow field-flow fractionation/multi angle laser light scattering. A uniform mechanistic hypothesis is presented to describe the impact of phospholipids on CXB permeation behavior: Obviously, the critical drug-to-phospholipid ratio represents a compromise between optimal stabilization of the amorphous state-induced supersaturation and reduced thermodynamic activity of CXB due to association with colloidal states, where the type of colloidal state (vesicle or micelle) appears to be of minor importance.</p

Moderate-to-High Intensity Physical Exercise in Patients with Alzheimer's Disease:A Randomized Controlled Trial

Background: Studies of physical exercise in patients with Alzheimer’s disease (AD) are few and results have been inconsistent. Objective: To assess the effects of a moderate-to-high intensity aerobic exercise program in patients with mild AD. Methods: In a randomized controlled trial, we recruited 200 patients with mild AD to a supervised exercise group (60-min sessions three times a week for 16 weeks) or to a control group. Primary outcome was changed from baseline in cognitive performance estimated by Symbol Digit Modalities Test (SDMT) in the intention-to-treat (ITT) group. Secondary outcomes included changes in quality of life, ability to perform activities of daily living, and in neuropsychiatric and depressive symptoms. Results: The ITT analysis showed no significant differences between intervention and control groups in change from baseline of SDMT, other cognitive tests, quality of life, or activities of daily living. The change from baseline in Neuropsychiatric Inventory differed significantly in favor of the intervention group (mean: –3.5, 95% confidence interval (CI) –5.8 to –1.3, p = 0.002). In subjects who adhered to the protocol, we found a significant effect on change from baseline in SDMT as compared with the control group (mean: 4.2, 95% CI 0.5 to 7.9, p = 0.028), suggesting a dose-response relationship between exercise and cognition. Conclusions: This is the first randomized controlled trial with supervised moderate-to-high intensity exercise in patients with mild AD. Exercise reduced neuropsychiatric symptoms in patients with mild AD, with possible additional benefits of preserved cognition in a subgroup of patients exercising with high attendance and intensity.</jats:p

Type 2 diabetes-related variants influence the risk of developing multiple myeloma: results from the IMMEnSE consortium

Type 2 diabetes (T2D) has been suggested to be a risk factor for multiple myeloma (MM), but the relationship between the two traits is still not well understood. The aims of this study were to evaluate whether 58 genome-wide-association-studies (GWAS)-identified common variants for T2D influence the risk of developing MM and to determine whether predictive models built with these variants might help to predict the disease risk. We conducted a case–control study including 1420 MM patients and 1858 controls ascertained through the International Multiple Myeloma (IMMEnSE) consortium. Subjects carrying the KCNQ1rs2237892T allele or the CDKN2A-2Brs2383208G/G, IGF1rs35767T/T and MADDrs7944584T/T genotypes had a significantly increased risk of MM (odds ratio (OR)=1.32–2.13) whereas those carrying the KCNJ11rs5215C, KCNJ11rs5219T and THADArs7578597C alleles or the FTOrs8050136A/A and LTArs1041981C/C genotypes showed a significantly decreased risk of developing the disease (OR=0.76–0.85). Interestingly, a prediction model including those T2D-related variants associated with the risk of MM showed a significantly improved discriminatory ability to predict the disease when compared to a model without genetic information (area under the curve (AUC)=0.645 vs AUC=0.629; P=4.05×10-06). A gender-stratified analysis also revealed a significant gender effect modification for ADAM30rs2641348 and NOTCH2rs10923931 variants (Pinteraction=0.001 and 0.0004, respectively). Men carrying the ADAM30rs2641348C and NOTCH2rs10923931T alleles had a significantly decreased risk of MM whereas an opposite but not significant effect was observed in women (ORM=0.71 and ORM=0.66 vs ORW=1.22 and ORW=1.15, respectively). These results suggest that TD2-related variants may influence the risk of developing MM and their genotyping might help to improve MM risk prediction models.This work was supported by grants from the FIBAO foundation (Granada, Spain) and the CRIS foundation against cancer, from the Cancer Network of Excellence (RD12/10 Red de Cancer) and from the Instituto de Salud Carlos III (Madrid, Spain; PI12/02688)

Hydrogenated phospholipid, a promising excipient in amorphous solid dispersions of fenofibrate for oral delivery:Preparation and in-vitro biopharmaceutical characterization

Amorphous solid dispersions (ASD) represent a viable formulation strategy to improve dissolution and bioavailability of poorly soluble drugs. Our study aimed to evaluate the feasibility and potential role of hydrogenated phospholipid (HPL) as a matrix material and solubilizing additive for binary (alone) or ternary (in combination with polymers) solid dispersions, using fenofibrate (FEN) as the model drug. FEN, incorporated within ASDs by melting or freeze-drying (up to 20% m/m), stayed amorphous during short-term stability studies. The solubility enhancing potential of HPL depended on the dissolution medium. In terms of enhancing in vitro permeation, solid dispersions with HPL were found equally or slightly more potent as compared to the polymer-based ASD. For studied ASD, in vitro permeation was found substantially enhanced as compared to a suspension of crystalline FEN and at least equal compared to marketed formulations under comparable conditions (literature data). Additionally, while the permeation of neat FEN and FEN in binary solid dispersions was affected by the dissolution medium (i.e., the “prandial state”), for ternary solid dispersions the permeation was independent of the “prandial state” (FaSSIF = FeSSIF). This suggests that ternary solid dispersions containing both polymer and HPL may represent a viable formulation strategy to mitigate fenofibrate's food effect.</p

A novel combined lipolysis-permeation screening approach in 96-well format:Method development using type I lipid-based formulations

For release testing of lipid-based formulations (LBFs), lipid digestion using the pH-stat approach is widely used despite the fact that it is a laborious exercise and predictivity towards in vivo performance could not be demonstrated. A probable reason is the lack of differentiation between readily absorbable (molecularly dissolved) and less absorbable (colloid associated) drug fractions. This work describes the development and testing of an alternative approach designed to address both issues of the pH-stat method, the labor-intensive nature and the limited ability to estimate in vivo behavior. The proposed solution involves combined lipolysis-permeation testing on 96-well microtiter sandwich plates. In this new time-efficient and material-sparing approach, a highly buffered lipolysis medium was used in the donor chamber to avoid the need for pH stabilization by titration. Moreover, the method was optimized to minimize non-specific adsorption of cinnarizine to the plates and polytetrafluoroethylene (PTFE)-coated stirring bars. The predictive power of the new high throughput screening (HTS) to estimate in vivo oral bioavailability of cinnarizine-loaded type I LBFs was evaluated against recent oral bioavailability data of the very same formulations in rats and compared to in vitro data generated by the pH-stat lipolysis approach. The following variables were studied: supersaturation, lipase inhibition, lipid chain length, and presence of an amphiphilic polymer (precipitation inhibitor). While the HTS method correctly captured the in vivo impact of both supersaturation and lipase inhibition for all formulations, the pH-stat method revealed opposite trends for one out of four combinations in each formulation sets. In vivo there had been no effect observed for lipid chain length nor presence of the amphiphilic polymer. In contrast, both in vitro approaches wrongly predicted such effects in some cases. A better prediction for the long-chain systems was found with the HTS method as with the laborious pH-stat approach. The HTS lipolysis-permeation method can test multiple formulations in the 96-well plate format within hours and gave IVIVCs of up to 0.91 for grouped type I LBFs. In particular the in vivo performance of supersaturated formulations was correctly captured. This study demonstrates that this new method represents a promising alternative to existing tools for prediction of the in vivo performance of type I LBFs.</p

Identification of miRSNPs associated with the risk of multiple myeloma

Accepted articleMultiple myeloma (MM) is a malignancy of plasma cells usually infiltrating the bone marrow, associated with the production of a monoclonal immunoglobulin (M protein) which can be detected in the blood and/or urine. Multiple lines of evidence suggest that genetic factors are involved in MM pathogenesis, and several studies have identified single nucleotide polymorphisms (SNPs) associated with the susceptibility to the disease. SNPs within miRNA-binding sites in target genes (miRSNPs) may alter the strength of miRNA-mRNA interactions, thus deregulating protein expression. MiRSNPs are known to be associated with risk of various types of cancer, but they have never been investigated in MM. We performed an in silico genome-wide search for miRSNPs predicted to alter binding of miRNAs to their target sequences. We selected 12 miRSNPs and tested their association with MM risk. Our study population consisted of 1,832 controls and 2,894 MM cases recruited from seven European countries and Israel in the context of the IMMEnSE (International Multiple Myeloma rESEarch) consortium. In this population two SNPs showed an association with p<0.05: rs286595 (located in gene MRLP22) and rs14191881 (located in gene TCF19). Results from IMMEnSE were meta-analyzed with data from a previously published genome-wide association study (GWAS). The SNPs rs13409 (located in the 3UTR of the POU5F1 gene), rs1419881 (TCF19), rs1049633, rs1049623 (both in DDR1) showed significant associations with MM risk. In conclusion, we sought to identify genetic polymorphisms associated with MM risk starting from genome-wide prediction of miRSNPs. For some mirSNPs, we have shown promising associations with MM risk. What's new? Even though deregulation of miRNA expression has been associated with human cancers little information is available regarding their relation with MM susceptibility. We performed an in silico genome-wide search for miRSNPs and selected the most promising ones for an association study. The SNPs with the strongest associations with MM risk are localized in genes which have never been related with MM.This work was partially funded by: intramural funds of German Cancer Research Center (DKFZ), Grant ref. HUS412A1271 from the “Gerencia Regional de Salud de la Junta de Castilla y Léon”. This work was supported by grants from the Instituto de Salud Carlos III (Madrid, Spain; PI12/02688). Catalan Government DURSI grant 2014SGR647 and Instituto de Salud Carlos III, co7funded by FEDER funds –a way to build Europe– grants PI11701439 and PIE13/00022info:eu-repo/semantics/publishedVersio

Enabling meta-analysis in systematic reviews on carpal tunnel syndrome

Possible solutions to the problems of clinical heterogeneity of outcome measures and inadequate reporting of results for randomized controlled trials (RCTs) on carpal tunnel syndrome (CTS) are presented. Meta-analysis was impeded by these problems in 2 systematic reviews concerning conservative and surgical treatment options for CTS. A solution to the problem of inadequate data presentation is to add explicit information on minimal requirements with regard to data presentation to guidelines for the reporting of studies. To resolve the problem of clinical heterogeneity of the outcomes there should be consensus on the (validated) outcomes that should be used in RCTs. For CTS there is little evidence available on the reliability, validity, and responsiveness to change of the commonly used outcomes in RCTs. Resolving both problems will increase the comparability of RCTs, enabling the calculation of a pooled estimate of effect in a meta-analysi

The association between Open Dialogue to young Danes in acute psychiatric crisis and their use of health care and social services:A retrospective register-based cohort study

Background: Although most mental disorders have their onset in early life, the mental health needs of young people are often not addressed adequately. Open Dialogue is a need-adapted approach that mobilizes psychosocial resources in a crisis struck person's social network. Open Dialogue is organised as a series of network meetings and seeks to promote collaborative integrated care, and a non-directive psychotherapeutic stance. Its effectiveness for young people has not previously been assessed. Objectives: The aim of the study was to examine whether a Danish Open Dialogue approach directed at young people, who sought help from Child and Adolescent Mental Health Services, reduced their utilisation of psychiatric and other health services, compared to peers receiving usual psychiatric treatment. Design: A retrospective register-based cohort study. Methods: Using clinical and national register data, a cohort of patients aged 14–19 years (n = 503) enrolled from one region during 2000 to 2015 were compared to a matched comparison group from two other regions using propensity scores. Utilisation of psychiatric health services, GP services, and social markers were assessed after 1, 2, 5 and 10 year of follow-up using logistic and Poisson regression models. Results: Patients receiving Open Dialogue intervention had more psychiatric outpatient treatments at one year of follow-up (RR = 1.2, CI: 1.1–1.4) than the comparison group, but not at subsequent follow-ups. Recipients of the intervention had fewer emergency psychiatric treatments (1 year follow-up: RR = 0.2, CI: 0.1-0.5; 10 years follow-up: RR = 0.5, CI: 0.3-0.8) and less use of general practitioner services (1 year follow-up: RR = 0.90, CI: 0.82-0.99; 10 years follow-up: RR = 0.85, CI: 0.78-0.92). There was no significant reduction in the number of psychiatric hospitalisation contacts or treatment days. Conclusions: Open Dialogue was significantly associated with some reduced risks of utilising health care services. These mixed results should be tested in a randomized design.</p