Fast-dissolving sublingual solid dispersion and cyclodextrin complex increase the absorption of perphenazine in rabbits

Objectives The sublingual administration route as well as solid dispersion formation with macrogol 8000 and complexation with β‐cyclodextrin (β‐CyD) were investigated as ways for improving the absorption of perphenazine, a poorly water‐soluble drug subjected to substantial first‐pass metabolism. Methods The absorption of perphenazine was studied in rabbits after sublingual administration of perphenazine/macrogol solid dispersion, solid perphenazine/β‐CyD complex and plain micronized perphenazine, as well as after peroral administration of an aqueous perphenazine solution. Solid formulations were prepared by freeze‐drying (perphenazine/macrogol solid dispersion) or spray‐drying (perphenazine/β‐CyD complex). Key findings The value for area under the curve from 0 to 360 min (AUC_{0–360 min}) of perphenazine after peroral administration was only 8% of the AUC_{0–360 min} value obtained after intravenous administration, while the corresponding values for the sublingually administered formulations were 53% (perphenazine/macrogol solid dispersion), 41% (perphenazine/β‐CyD complex) and 64% (micronized perphenazine). There are three possible mechanisms to explain these results: avoidance of the first‐pass metabolism; good sublingual absorption of perphenazine; and rapid dissolution rate of perphenazine from the studied formulations. Conclusions With sublingual administration, the drug has to dissolve rapidly in a small volume of saliva. Based on the present absorption studies in rabbits, the solid dispersion preparation and cyclodextrin complexation were postulated to be useful ways to attain successful sublingual administration of perphenazine. Good sublingual absorption was also achieved by micronization of perphenazine. As far as we are aware, this paper is one of the first to evaluate the sublingual administration of a solid dispersion in vivo.     

Glycodelin expression associates with differential tumour phenotype and outcome in sporadic and familial non-<Emphasis Type="Italic">BRCA1</Emphasis>/<Emphasis Type="Italic">2</Emphasis> breast cancer patients

Glycodelin (encoded by PAEP gene) is a secreted lipocalin protein mainly expressed in reproductive tissues, but also in several tumour types. In the breast, glycodelin is expressed both in normal epithelial and cancerous tissue. To investigate the association of glycodelin with clinicopathological features of breast cancer and outcome of patients we evaluated the protein expression of glycodelin in a large series of breast tumours. Immunohistochemical analysis of tissue microarrays was used to study glycodelin expression on 399 sporadic and 436 familial non-BRCA1/2 tumours with strong family history. Gene expression analysis was used to define genes co-expressed with PAEP in sporadic and familial non-BRCA1/2 breast tumours. In the sporadic series, the glycodelin expression associated with low proliferation rate (P < 0.001), with a tendency towards well-differentiated tumours (grades 1 and 2, P = 0.012) and high cyclin D1 (P = 0.034) expression. However, in familial non-BRCA1/2 cases with strong family history glycodelin expression associated with a less favourable phenotype, i.e. positive lymph node status (P = 0.003) and HER2-positive tumours (P = 0.009). Moreover, the patients with glycodelin-positive tumours had an increased risk for distant metastases (P = 0.001) and in multivariate analysis glycodelin expression was an independent predictor of metastasis (hazard ratio (HR) = 2.22, 95% confidence interval (95% CI) = 1.22–4.03, P = 0.009) in familial non-BRCA1/2 breast cancer. Gene expression analysis further revealed different gene expression profiles correlating with the PAEP expression in the sporadic and familial non-BRCA1/2 breast cancers. Our findings suggest differential progression pathways in the sporadic and familial non-BRCA1/2 breast tumours expressing glycodelin.     

Immobilization of protein-coated drug nanoparticles in nanofibrillar cellulose matrices—Enhanced stability and release

Nanosizing is an advanced approach to overcome poor aqueous solubility of active pharmaceutical ingredients. One main problem in pharmaceutical nanotechnology is maintaining of the morphology of the nanometer sized particles during processing and storage to make sure the formulation behaves as originally planned. Here, a genetically engineered hydrophobin fusion protein, where the hydrophobin (HFBI) was coupled with two cellulose binding domains (CBDs), was employed in order to facilitate drug nanoparticle binding to nanofibrillar cellulose (NFC). The nanofibrillar matrix provides protection for the nanoparticles during the formulation process and storage. It was demonstrated that by enclosing the functionalized protein coated itraconazole nanoparticles to the external nanofibrillar cellulose matrix notably increased their storage stability. In a suspension with cellulose nanofibrils, nanoparticles around 100 nm could be stored for more than ten months when the specific cellulose binding domain was fused to the hydrophobin. Also freeze-dried particles in the cellulose nanofibrils matrix were preserved without major changes in their morphology. In addition, as a consequence of formation of the immobilized nanodispersion, dissolution rate of itraconazole was increased significantly, which also enhanced the in vivo performance of the drug.     

Arachidonic acid pathway members PLA2G7, HPGD, EPHX2, and CYP4F8 identified as putative novel therapeutic targets in prostate cancer

The arachidonic acid and prostaglandin pathway has been implicated in prostate carcinogenesis, but comprehensive studies of the individual members in this key pathway are lacking. Here, we first conducted a systematic bioinformatic study of the expression of 36 arachidonic acid pathway genes across 9783 human tissue samples. The results showed that the PLA2G7, HPGD, EPHX2, and CYP4F8 genes are highly expressed in prostate cancer. Functional studies using RNA interference in prostate cancer cells indicated that all four genes are also essential for cell growth and survival. Clinical validation confirmed high PLA2G7 expression, especially in ERG oncogene-positive prostate cancers, and its silencing sensitized ERG-positive prostate cancer cells to oxidative stress. HPGD was highly expressed in androgen receptor (AR)-overexpressing advanced tumors, as well as in metastatic prostate cancers. EPHX2 mRNA correlated with AR in primary prostate cancers, and its inhibition in vitro reduced AR signaling and potentiated the effect of antiandrogen flutamide in cultured prostate cancer cells. In summary, we identified four novel putative therapeutic targets with biomarker potential for different subtypes of prostate cancer. In addition, our results indicate that inhibition of these enzymes may be particularly powerful when combined with other treatments, such as androgen deprivation or induction of oxidative stress.     

Impaired cognitive inhibition in schizophrenia: a meta-analysis of the Stroop interference effect

Schizophrenia has been consistently shown to be associated with impairment in executive functioning. However, although frequently treated as such, the term executive functioning does not refer to a unitary cognitive function; it rather represents a set of basic, lower-level cognitive sub-components, e.g. updating, shifting, and cognitive inhibition. This specification into sub-components allows for a further differentiation of the executive deficits found in schizophrenia. Focusing on the sub-component of cognitive inhibition, we here present a meta-analysis of interference effect as assessed with the Stroop Color-Word Interference paradigm. Including the results of 36 studies with 1081 schizophrenia patients and 1026 healthy control subjects, it was shown that schizophrenia patients exhibit an increased Stroop interference effect both in response time (mean effect size: M(g) = 0.43; 95% confidence interval, CI95%: 0.35-0.52) and accuracy (M(g) = 0.62; CI95%: 0.47-0.77) measures of interference. However, a meta-regression analysis revealed that the size of the effect varies depending on the version of the Stroop paradigm used. Regarding the response time measures of interference, studies using the classical card version of the paradigm showed a significantly larger effect size than studies using a single-trial computerized version of the paradigm (M(g) = 0.60 vs. M(g) = 0.19). Despite of the dissociation between the two versions of the paradigm, the results of the present meta-analysis indicate that the reported global deficits in executive functioning found to be associated with schizophrenia are at least partly due to a reduced ability of cognitive inhibition.     

Food frequency questionnaires and overnight urines are valid indicators of daidzein and genistein intake in U.S. women relative to multiple 24-h urine samples

Data regarding convenient, valid methods for measuring U.S. isoflavone intake are limited. We evaluated a soy food questionnaire (SFQ), the Willett food frequency questionnaire (FFQ), and overnight urine samples relative to excretion in 24-h urine samples. We also described intake among women in a high-risk program for breast or ovarian cancer. Between April 2002 and June 2003, 451 women aged 30 to 50 yr with a family history of breast or ovarian cancer completed the SFQ and FFQ. Of them, 27 provided four 24-h and overnight urine specimens. In these women, 24-h sample measures were correlated with SFQ estimates of daidzein (Spearman r = .48) and genistein (r = .54) intake, moderately correlated with the Willett FFQ (daidzein r = .38, genistein r = .33), and strongly correlated with overnight urine excretion (daidzein r = .84, genistein r = 0.93). Among all 451 SFQ respondents, mean (median) daidzein and genistein intakes were 2.8 (0.24) and 3.9 (0.30) mg/day. Primary sources of both were soymilk, soy nuts, and tofu. We conclude that targeted soy food questionnaires, comprehensive FFQs, and multiple overnight urines are all reasonable options for assessing isoflavone intake in epidemiologic studies.     

Genome-wide search for breast cancer linkage in large Icelandic non-BRCA1/2 families

Introduction:

A significant proportion of high-risk breast cancer families are not explained by mutations in known genes. Recent genome-wide searches (GWS) have not revealed any single major locus reminiscent of BRCA1 and BRCA2, indicating that still unidentified genes may explain relatively few families each or interact in a way obscure to linkage analyses. This has drawn attention to possible benefits of studying populations where genetic heterogeneity might be reduced. We thus performed a GWS for linkage on nine Icelandic multiple-case non-BRCA1/2 families of desirable size for mapping highly penetrant loci. To follow up suggestive loci, an additional 13 families from other Nordic countries were genotyped for selected markers.

Methods:

GWS was performed using 811 microsatellite markers providing about five centiMorgan (cM) resolution. Multipoint logarithm of odds (LOD) scores were calculated using parametric and nonparametric methods. For selected markers and cases, tumour tissue was compared to normal tissue to look for allelic loss indicative of a tumour suppressor gene.

Results:

The three highest signals were located at chromosomes 6q, 2p and 14q. One family contributed suggestive LOD scores (LOD 2.63 to 3.03, dominant model) at all these regions, without consistent evidence of a tumour suppressor gene. Haplotypes in nine affected family members mapped the loci to 2p23.2 to p21, 6q14.2 to q23.2 and 14q21.3 to q24.3. No evidence of a highly penetrant locus was found among the remaining families. The heterogeneity LOD (HLOD) at the 6q, 2p and 14q loci in all families was 3.27, 1.66 and 1.24, respectively. The subset of 13 Nordic families showed supportive HLODs at chromosome 6q (ranging from 0.34 to 1.37 by country subset). The 2p and 14q loci overlap with regions indicated by large families in previous GWS studies of breast cancer.

Conclusions:

Chromosomes 2p, 6q and 14q are candidate sites for genes contributing together to high breast cancer risk. A polygenic model is supported, suggesting the joint effect of genes in contributing to breast cancer risk to be rather common in non-BRCA1/2 families. For genetic counselling it would seem important to resolve the mode of genetic interaction.