Nonsteroidal Androgen Receptor Ligands: Versatile Syntheses and Biological Data

We report herein a stereoselective and straightforward methodology for the synthesis of new androgen receptor ligands with (anti)-agonistic activities. Oxygen–nitrogen replacement in bicalutamide-like structures paves the way to the disclosure of a new class of analogues, including cyclized/nitrogen-substituted derivatives, with promising antiandrogen (or anabolic) activity

Study of molecular mechanisms of pro-apoptotic activity of NCX 4040, a novel nitric oxide-releasing aspirin, in colon cancer cell lines-4

<p><b>Copyright information:</b></p><p>Taken from "Study of molecular mechanisms of pro-apoptotic activity of NCX 4040, a novel nitric oxide-releasing aspirin, in colon cancer cell lines"</p><p>http://www.translational-medicine.com/content/5/1/52</p><p>Journal of Translational Medicine 2007;5():52-52.</p><p>Published online 30 Oct 2007</p><p>PMCID:PMC2174440.</p><p></p>tibody that specifically recognizes the presence of nuclear 8-hydroxyguanine lesions, which are almost exclusively elicited by oxidative stress. Each 100× magnification shows a detail of the corresponding 40× photo. All the pictures are representative of three independent experiments

Study of molecular mechanisms of pro-apoptotic activity of NCX 4040, a novel nitric oxide-releasing aspirin, in colon cancer cell lines-0

<p><b>Copyright information:</b></p><p>Taken from "Study of molecular mechanisms of pro-apoptotic activity of NCX 4040, a novel nitric oxide-releasing aspirin, in colon cancer cell lines"</p><p>http://www.translational-medicine.com/content/5/1/52</p><p>Journal of Translational Medicine 2007;5():52-52.</p><p>Published online 30 Oct 2007</p><p>PMCID:PMC2174440.</p><p></p>μM, followed by a 24-hour wash-out. Growth inhibition and cytocidal effect of drugs were calculated according to Monks' formula, as reported in Materials and methods. Each point indicates the mean of at least three experiments; SD never exceeded 5%

R a 24-hour exposure to 10 μM and 50 μM of NCX 4040, respectively, as evidenced by DAPI staining

<p><b>Copyright information:</b></p><p>Taken from "Study of molecular mechanisms of pro-apoptotic activity of NCX 4040, a novel nitric oxide-releasing aspirin, in colon cancer cell lines"</p><p>http://www.translational-medicine.com/content/5/1/52</p><p>Journal of Translational Medicine 2007;5():52-52.</p><p>Published online 30 Oct 2007</p><p>PMCID:PMC2174440.</p><p></p> . Genomic DNA isolated from LoVo () and LRWZ () cells after different exposure times to NCX 4040 was electrophoresed on 1.5 % agarose gel to detect internucleosomal DNA fragmentation. () (10-μM concentration of NCX 4040): lane a, untreated; lane b, 2-hour exposure; lane c, 4-hour exposure; lane d, 6-hour exposure; lane e, 24-hour exposure. () (50-μM concentration of NCX 4040): lane a, untreated; lane b, 2-hour exposure; lane c, 4-hour exposure; lane d, 8-hour exposure; lane e, 16-hour exposure; lane f, 24-hour exposure; lane g, 48-hour exposure

Study of molecular mechanisms of pro-apoptotic activity of NCX 4040, a novel nitric oxide-releasing aspirin, in colon cancer cell lines-3

<p><b>Copyright information:</b></p><p>Taken from "Study of molecular mechanisms of pro-apoptotic activity of NCX 4040, a novel nitric oxide-releasing aspirin, in colon cancer cell lines"</p><p>http://www.translational-medicine.com/content/5/1/52</p><p>Journal of Translational Medicine 2007;5():52-52.</p><p>Published online 30 Oct 2007</p><p>PMCID:PMC2174440.</p><p></p>after different exposure times to NCX 4040. () Cytochrome c, caspase-9 and -3 protein expression in LoVo cells: lane a, untreated; lane b, 6-hour NCX 4040 exposure (10 μM). () Cytochrome c, caspase-9 and -3 protein expression in LRWZ cells: lane a, untreated; lane b, 14-hour NCX 4040 exposure (50 μM)

Study of molecular mechanisms of pro-apoptotic activity of NCX 4040, a novel nitric oxide-releasing aspirin, in colon cancer cell lines-5

<p><b>Copyright information:</b></p><p>Taken from "Study of molecular mechanisms of pro-apoptotic activity of NCX 4040, a novel nitric oxide-releasing aspirin, in colon cancer cell lines"</p><p>http://www.translational-medicine.com/content/5/1/52</p><p>Journal of Translational Medicine 2007;5():52-52.</p><p>Published online 30 Oct 2007</p><p>PMCID:PMC2174440.</p><p></p>of apoptotic cells after exposure of WiDr and LRWZ cells to NS398, NCX 4040 or both drugs

Study of molecular mechanisms of pro-apoptotic activity of NCX 4040, a novel nitric oxide-releasing aspirin, in colon cancer cell lines-6

<p><b>Copyright information:</b></p><p>Taken from "Study of molecular mechanisms of pro-apoptotic activity of NCX 4040, a novel nitric oxide-releasing aspirin, in colon cancer cell lines"</p><p>http://www.translational-medicine.com/content/5/1/52</p><p>Journal of Translational Medicine 2007;5():52-52.</p><p>Published online 30 Oct 2007</p><p>PMCID:PMC2174440.</p><p></p>d LoVo Dx cells were analyzed by real-time RT-PCR

Study of molecular mechanisms of pro-apoptotic activity of NCX 4040, a novel nitric oxide-releasing aspirin, in colon cancer cell lines-2

<p><b>Copyright information:</b></p><p>Taken from "Study of molecular mechanisms of pro-apoptotic activity of NCX 4040, a novel nitric oxide-releasing aspirin, in colon cancer cell lines"</p><p>http://www.translational-medicine.com/content/5/1/52</p><p>Journal of Translational Medicine 2007;5():52-52.</p><p>Published online 30 Oct 2007</p><p>PMCID:PMC2174440.</p><p></p>dye JC-1 and flow cytometric analysis. JC-1 exhibits potential-dependent accumulation in mitochondria, as indicated by a fluorescence emission shift from green to red. ΔΨdepolarization is indicated by a decrease in the red/green fluorescence intensity ratio, which is dependent only on the membrane potential and not on other factors such as mitochondrial size, shape or density. FL2-H, median red fluorescence intensity; h, hour

Effect of Small Molecules Modulating Androgen Receptor (SARMs) in Human Prostate Cancer Models

<div><p>The management of hormone-refractory prostate cancer represents a major challenge in the therapy of this tumor, and identification of novel androgen receptor antagonists is needed to render treatment more effective. We analyzed the activity of two novel androgen receptor antagonists, (<i>S</i>)-<b>11</b> and (<i>R</i>)-<b>9</b>, in <i>in vitro</i> and <i>in vivo</i> experimental models of hormone-sensitive or castration-resistant prostate cancer (CRPC). <i>In vitro</i> experiments were performed on LNCaP, LNCaP-AR, LNCaP-Rbic and VCaP human prostate cancer cells. Cytotoxic activity was assessed by SRB and BrdU uptake, AR transactivation by luciferase reporter assay and PSA levels by Real Time RT-PCR and ELISA assays. Cell cycle progression-related markers were evaluated by western blot. <i>In vivo</i> experiments were performed on SCID mice xenografted with cells with different sensitivity to hormonal treatment. In hormone-sensitive LNCaP and LNCaP-AR cells, the latter expressing high androgen receptor levels, (<i>R</i>)-<b>9</b> and (<i>S</i>)-<b>11</b> exhibited a higher cytotoxic effect compared to that of the reference compound ((<i>R</i>)-bicalutamide), also in the presence of the synthetic androgen R1881. Furthermore, the cytotoxic effect produced by (<i>R</i>)-<b>9</b> was higher than that of (<i>S</i>)-<b>11</b> in the two hormone-resistant LNCaP-AR and VCaP cells. A significant reduction in PSA levels was observed after exposure to both molecules. Moreover, (<i>S</i>)-<b>11</b> and (<i>R</i>)-<b>9</b> inhibited DNA synthesis by blocking the androgen-induced increase in cyclin D1 protein levels. <i>In vivo</i> studies on the toxicological profile of (<i>R</i>)-<b>9</b> did not reveal the presence of adverse events. Furthermore, (<i>R</i>)-<b>9</b> inhibited tumor growth in various <i>in vivo</i> models, especially LNCaP-Rbic xenografts, representative of recurrent disease. Our <i>in vitro</i> results highlight the antitumor activity of the two novel molecules (<i>R</i>)-<b>9</b> and (<i>S</i>)-<b>11</b>, making them a potentially attractive option for the treatment of CRPC.</p></div

Structure compounds and their AR-specific binding properties.

<p>(<b>A</b>) Chemical structure and molecular weight (m.w.) of (<i>R</i>)-bicalutamide, (<i>S</i>)-11 and (<i>R</i>)-9. (<b>B</b>) AR ligand binding displacement analysis in LNCaP cells<b>.</b> Quiescent LNCaP cells were incubated with 10 nM [3H] R1881 in the absence or presence of the indicated excess (from 0.5 µM to 4 µM) of radio inert compounds. Intracellular radioactivity was assayed. Inset in panel B shows the AR binding sites assayed in 10³ cells incubated with 10 nM of [3H] R1881 (R1881*) in the absence or presence of the indicated excess of unlabeled (<i>R</i>)-<b>9</b>, (<i>S</i>)-<b>11</b> or (<i>R</i>)-bicalutamide (<i>R</i>)-bic. Data from three different experiments were collected and residual binding was calculated and expressed as % of total AR binding sites. <i>n</i> =  number of experiments. The statistical significance of results was also evaluated by the paired <i>t</i> test and P values <0.005 were considered significant. No significance was attributed to the difference in the residual binding between the cells incubated with 10 nM [3H] R1881 in the presence of (<i>S</i>)-11 or (<i>R</i>)-9 and those incubated with 10 nM [3H] R1881 in the presence of (<i>R</i>)-bicalutamide ((<i>R</i>)-bic). LNCaP cells were also incubated with 10 nM [3H] R1881 in the absence or presence of 100-fold excess (1 µM) of unlabeled R1881 or Casodex®. Residual binding was 13% and 14% for unlabeled R1881 or Casodex®, respectively.</p