Effect of chronic treatment with Rosiglitazone on Leydig cell steroidogenesis in rats: In vivo and ex vivo studies

<p>Abstract</p> <p>Background</p> <p>The present study was designed to examine the effect of chronic treatment with rosiglitazone - thiazolidinedione used in the treatment of type 2 diabetes mellitus for its insulin sensitizing effects - on the Leydig cell steroidogenic capacity and expression of the steroidogenic acute regulatory protein (StAR) and cholesterol side-chain cleavage enzyme (P450scc) in normal adult rats.</p> <p>Methods</p> <p>Twelve adult male Wistar rats were treated with rosiglitazone (5 mg/kg) administered by gavage for 15 days. Twelve control animals were treated with the vehicle. The ability of rosiglitazone to directly affect the production of testosterone by Leydig cells <it>ex vivo </it>was evaluated using isolated Leydig cells from rosiglitazone-treated rats. Testosterone production was induced either by activators of the cAMP/PKA pathway (hCG and dbcAMP) or substrates of steroidogenesis [22(R)-hydroxy-cholesterol (22(R)-OH-C), which is a substrate for the P450scc enzyme, and pregnenolone, which is the product of the P450scc-catalyzed step]. Testosterone in plasma and in incubation medium was measured by radioimmunoassay. The StAR and P450scc expression was detected by immunocytochemistry.</p> <p>Results</p> <p>The levels of total circulating testosterone were not altered by rosiglitazone treatment. A decrease in basal or induced testosterone production occurred in the Leydig cells of rosiglitazone-treated rats. The ultrastructural and immunocytochemical analysis of Leydig cells from rosiglitazone-treated rats revealed cells with characteristics of increased activity as well as increased StAR and P450scc expression, which are key proteins in androgen biosynthesis. However, a number of rosiglitazone-treated cells exhibited significant mitochondrial damage.</p> <p>Conclusion</p> <p>The results revealed that the Leydig cells from rosiglitazone-treated rats showed significant reduction in testosterone production under basal, hCG/dbcAMP- or 22 (R)-OH-C/pregnenolone-induced conditions, although increased labeling of StAR and P450scc was detected in these cells by immunocytochemistry. The ultrastructural study suggested that the lower levels of testosterone produced by these cells could be due to mitochondrial damage induced by rosiglitazone.</p

2-(2-Nitro­anilino)-4,5,6,7-tetra­hydro­benzo[b]thio­phene-3-carbonitrile

The title compound, C15H13N3O2S, was synthesized by the reaction of 2-amino-5,6,7,8-tetra­hydro-4H-cyclo­hepta­[b]thio­phene-3-carbonitrile and o-fluoro­nitro­benzene. The dihedral angle between the thio­phene and nitro­phenyl rings is 75.15 (2)°. In the crystal, inter­molecular N—H⋯N and C—H⋯O inter­actions lead to the formation of a supra­molecular chain extending along the c-axis direction

2-Amino-4,5,6,7-tetra­hydro­benzo[b]thio­phene-3-carbonitrile

The title compound, C9H10N2S, was synthesized according to Gewald procedures by the reaction of cyclo­hexa­none with malonitrile and sulfur in the presence morpholine. The cyclo­hexane ring adopts a half-chair conformation and the thio­phene ring is essentially planar (r.m.s. deviation = 0.05 Å). The crystal packing is stabilized by two inter­molecular N—H⋯N hydrogen bonds, which link the mol­ecules into centrosymmetric rings with graph-set motif R 2 2(12)

2-(2-Nitro­anilino)-5,6,7,8-tetra­hydro-4H-cyclo­hepta­[b]thio­phene-3-carbonitrile

The title compound, C16H15N3O2S, was synthesized by the reaction of 2-amino-5,6,7,8-tetra­hydro-4H-cyclo­hepta­[b]thio­phene-3-carbonitrile and o-fluoro­nitro­benzene. The thio­phene and nitro­phenyl rings and amino and carbonitrile groups are coplanar with a maximum deviation of 0.046 (2) Å and a dihedral angle of 0.92 (6)° between the rings. The cyclo­hepta ring adopts a chair conformation. Intra­molecular N—H⋯O and C—H⋯S inter­actions occur. In the crystal, the mol­ecules form layers that are linked by π–π stacking inter­actions between the thio­phene and benzene rings [centroid–centroid distances = 3.7089 (12) and 3.6170 (12) Å]

Synthesis and evaluation of the antifungal activity of 2-(substituted-amino)-4,5-dialkyl-thiophene-3- carbonitrile derivatives

Fifteen 2-[(substituted-benzylidene)-amino]-5-methyl-thiophene-3-carbonitrile (3a-g) and 2-[(substituted-benzylidene)-amino]-4,5-cycloalkyl-thiophene-3-carbonitrile derivatives (4a-h) were synthesized and screened for their in vitro antifungal activity against 42 clinical isolates of Candida (representing 4 different species) and 2 isolates of Criptococcus. The antifungal activities of these compounds were compared to fluconazole and amphotericin B as standard agents. All compounds presented fungicidal activity at different doses, but a few compounds showed moderate or poor antifungal activity when compared with the standard drugs. The Cryptococcus strains were more sensitive than those of the genus Candida, and compound 4d was the most active, with MFC values varying between 100-800 μg/mL. A preliminary SAR study demonstrated that the presence of a cycloalkyl ring linked to the thiophene moiety is essential for antifungal activity, and that the best antifungal candidates are cyclohexyl compounds (4d-f). The results suggest that thiophene derivatives may be interesting compounds for the further development of antifungal drugs.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Preliminarno ispitivanje antimikotskog i citotoksičnog djelovanja derivata cikloalkil[b]tiofena PLS-DA analizom

A series of 2-[(arylidene)amino]-cycloalkyl[b]thiophene-3-carbonitriles (2a-x) was synthesized by incorporation of substituted aromatic aldehydes in Gewald adducts (1a-c). The title compounds were screened for their antifungal activity against Candida krusei and Criptococcusneoformans and for their antiproliferative activity against a panel of 3 human cancer cell lines (HT29, NCI H-292 and HEP). Forantiproliferative activity, the partial least squares (PLS) methodology was applied. Some of the prepared compounds exhibited promising antifungal and proliferative properties. The most active compounds for antifungal activity were cyclohexyl[b]thiophene derivatives, and forantiproliferative activity cycloheptyl[b]thiophene derivatives, especially 2-[(1H-indol-2-yl-methylidene)amino]-5,6,7,8-tetrahydro-4H-cyclohepta[b]thiophene-3-carbonitrile (2r), which inhibited more than 97 % growth of the three cell lines. The PLS discriminant analysis (PLS-DA)applied generated good exploratory and predictive results and showed that the descriptors having shape characteristics were strongly correlated with the biological data.Koristeći supstituirane aromatske aldehide u Gewaldovim aduktima 1a-c sintetizirani su derivati 2-[(ariliden)amino]-cikloalkil[b]tiofen-3-karbonitrila (2a-x). Ispitano je antimikotsko djelovanje tih spojeva na gljivice Candida krusei i Criptococcus neoformans te antiproliferativno djelovanje na tri humane tumorske stanične linije (HT29, NCI H-292 i HEP). Za antiproliferativno djelovanje primijenjena je metoda parcijalnih najmanjih kvadrata (PLS) koristeći softverski program Pentacle. Neki od ispitanih spojeva pokazuju obećavajuće antimikotsko i antiproliferativno djelovanje. Najjače antimikotsko djelovanje imaju cikloheksil[b]tiofen derivati, a najjače antiproliferativno djelovanje cikloheptil[b]tiofen derivati, posebice 2-[(1H-indol-2-il-metiliden)amino]-5,6,7,8-tetrahidro-4H-ciklohepta[b]tiofen-3-karbonitril (2r), koji inhibira više od 97 % rast svih triju ispitivanih staničnih linija. Primijenjena PLS diskriminirajuća analiza dala je dobre istraživačke i prognostičke rezultate i pokazala da deskriptori dobro koreliraju s biološkim rezultatima

Photodynamic antimicrobial chemotherapy (PACT) using phenothiazine derivatives as photosensitizers against Leishmania braziliensis

Background and Objective Cutaneous and mucocutaneous leishmaniasis are diseases characterized by skin or mucosal manifestations. In the new world, Leishmania braziliensis is the main etiological agent of cutaneous leishmaniasis, condition that may evolve to the mucocutaneous form. The therapeutic arsenal routinely employed to treat infected patients is unsatisfactory, especially for pentavalent antimonials, treatment recommended by the WHO, as they are often highly toxic, poorly tolerated and of variable effectiveness. This work aimed to evaluate in vitro the effectiveness of photodynamic antimicrobial chemotherapy as a new approach for the treatment of leishmaniasis. Materials and Methods A laser (??=?660?nm, 40?mW, 4.2?J/cm2, and 8.4?J/cm2, CW) associated to phenothiazine's derivatives (5 and 10?mu g/ml, toluidine blue O, methylene blue, or phenothiazine) on the promastigote forms of L. braziliensis in a single session. Samples were removed and analyzed in a hemocytometer 72?hours after PACT and viability of the parasites was assessed in quadruplicates. Results An important decrease in the number of viable parasites on all treated groups in comparison to their controls was observed as all tested compounds lead to significant parasite lethality being the highest lethality achieved with 10?mu g/ml of TBO. No lethality was observed on groups treated with laser or with any of the compounds separately. Conclusions TBO presented higher parasite lethality in comparison to MB with impressive reduction from 1?hour to 5?minutes of pre-incubation time. Lasers Surg. Med. 44: 850855, 2012. (c) 2012 Wiley Periodicals, Inc

Development and application of LC-UV method for the quantification of the anti-inflammatory thiazolidinone PG15 in rat plasma

A simple and rapid liquid chromatography-ultraviolet detection (LC-UV) method has been developed and validated for quantifying (5Z,E)-3-[2-(4-chlorophenyl)-2-oxoethyl]-5-(1H-indol-3-ylmethylene)-thiazolidine-2,4-dione (PG15) in rat plasma. A C18 reversed phase column provided chromatographic separation of the analyte which was followed by UV detection at 385 nm. The method involves precipitation of PG15 from plasma and isocratic elution with methanol:water (90:10, v/v). Total elution time was 7.5 min. The proposed method was validated and showed linear correlation in the range of 62.5 to 4000 ng mL-1. The within- and between-day precision, expressed as the relative standard error, were found to be less than 15 and 10 %, respectively, for all the concentrations investigated. The accuracy, measured using the quality control samples, was in the range of 86.1-114.9 %. The applicability of the validated method was tested in a pre-clinical pharmacokinetic study of the thiazolidinone PG15