Structure-Activity Relationships for the Marine Natural Product Sintokamides: Androgen Receptor N-Terminus Antagonists of Interest for Treatment of Metastatic Castration-Resistant Prostate Cancer.

Synthetic analogues of the marine natural product sintokamides have been prepared in order to investigate the structure-activity relationships for the androgen receptor N-terminal domain (AR NTD) antagonist activity of the sintokamide scaffold. An in vitro LNCaP cell-based transcriptional activity assay with an androgen-driven luciferase (Luc) reporter was used to monitor the potency of analogues. The data have shown that the chlorine atoms on the leucine side chains are essential for potent activity. Analogues missing the nonchlorinated methyl groups of the leucine side chains (C-1 and C-17) are just as active and in some cases more active than the natural products. Analogues with the natural R configuration at C-10 and the unnatural R configuration at C-4 are most potent. Replacing the natural propionamide N-terminus cap with the more sterically hindered pivaloylamide N-terminus cap leads to enhanced potency. The tetramic acid fragment and the methyl ether on the tetramic acid fragment are essential for activity. The SAR optimized analogue 76 is more selective, easier to synthesize, more potent, and presumed to be more resistant to proteolysis than the natural sintokamides.

Sintokamide A Is a Novel Antagonist of Androgen Receptor That Uniquely Binds Activation Function-1 in Its Amino-terminal Domain.

Androgen receptor (AR) is a validated drug target for all stages of prostate cancer including metastatic castration-resistant prostate cancer (CRPC). All current hormone therapies for CRPC target the C-terminal ligand-binding domain of AR and ultimately all fail with resumed AR transcriptional activity. Within the AR N-terminal domain (NTD) is activation function-1 (AF-1) that is essential for AR transcriptional activity. Inhibitors of AR AF-1 would potentially block most AR mechanisms of resistance including constitutively active AR splice variants that lack the ligand-binding domain. Here we provide evidence that sintokamide A (SINT1) binds AR AF-1 region to specifically inhibit transactivation of AR NTD. Consistent with SINT1 targeting AR AF-1, it attenuated transcriptional activities of both full-length AR and constitutively active AR splice variants, which correlated with inhibition of growth of enzalutamide-resistant prostate cancer cells expressing AR splice variants. In vivo, SINT1 caused regression of CRPC xenografts and reduced expression of prostate-specific antigen, a gene transcriptionally regulated by AR. Inhibition of AR activity by SINT1 was additive to EPI-002, a known AR AF-1 inhibitor that is in clinical trials (NCT02606123). This implies that SINT1 binds to a site on AF-1 that is unique from EPI. Consistent with this suggestion, these two compounds showed differences in blocking AR interaction with STAT3. This work provides evidence that the intrinsically disordered NTD of AR is druggable and that SINT1 analogs may provide a novel scaffold for drug development for the treatment of prostate cancer or other diseases of the AR axis.

Incidence of Diabetes Insipidus in Postoperative Period among the Patients Undergoing Pituitary Tumour Surgery.

Post operative complications after pituitary tumour surgery vary according to procedure. There are several surgical procedures being done such as transcranial, transsphenoidal microsurgical and transsphenoidal endoscopic approaches. One of the commonest complications is diabetes insipidus (DI). Our main objective was to find out the incidence of diabetes insipidus in post operative period among patients undergoing surgical intervention for pituitary tumour in our institute. The presence of diabetes insipidus in the postoperative period was established by measuring serum Na+ concentration, hourly urine output and urinary specific gravity to find out the incidence of diabetes insipidus in postoperative period in relation to age, gender, tumour diameter, function of tumour (i.e., either hormone secreting or not) and operative procedure used for surgical resection of pituitary tumor. As it is the most common postoperative complication so, in this study we tried to find out how many of the patients develop diabetes insipidus in postoperative period following surgical resection of pituitary tumour. This cross sectional type of observational study was carried out in the department of Neurosurgery, BSMMU from May 2014 to October 2015 on 33 consecutive patients who underwent surgical intervention for pituitary tumour for the first time. Data was collected by using a data collection sheet. The incidence of diabetes insipidus was found 23.1% of patients in <30 year age group, 38.5% of patients in 31-40 year age group and 38.5% of patients in ≥40 year age group (p=0.764). In case of distribution of patients according to gender 38.5% of male and 61.5% of female developed diabetes insipidus (p=0.073). Regarding tumour size 30.8% and 69.2% of patients developed diabetes insipidus having tumour diameter <30mm and ≥30mm respectively (p=0.590). In case of operative procedure 69.2% of patients developed diabetes insipidus who was operated by transsphenoidal endoscopic approach, 23.1% and 7.7% of patients developed diabetes insipidus who underwent pituitary tumour resection through transsphenoidal microscopic approach and transcranial microscopic approach respectively (p=0.432). 17.6% of patients develop DI having functioning pituitary macroadenoma and 62.5% of patients develop DI having nonfunctioning pituitary macroadenoma. This observational study has been performed to find out the incidence of diabetes insipidus. Incidence of postoperative DI is more at or around the age of 40 years. It is slightly predominant in female. Most of the patients manifest DI in the first 24 hours of surgical intervention. Incidence of DI is low among patients having functioning pituitary macroadenoma.

Efficiency upgrading of solar PVT finned hybrid system in Bangladesh: Flow rate and temperature influences.

This research aims to determine the impact of mass flow rate and inflow temperature on the utility and effectiveness of solar thermal systems using fins with air in various applications in Bangladesh. This study examines a three-dimensional (3D) photovoltaic thermal (PVT) system where we analyze the behavior of a hybrid system with six aluminum sheets (1 mm thick fin as a heat exchange material) inside the heat exchanger where the air takes the direction to pass in waveform through the channels (made of aluminum) using fins. The top side of the fins is bent and affixed to the bottom of the floor of the PV panel to allow heat transfer utilizing the conduction-based method. This study selects inlet fluid mass flow rate and inflow temperature between (0.015-0.535 kg/s), and (10-40 °C) respectively, while comparing the result with experimental/numerical published data based on Bangladesh's weather conditions and applies the finite element method (FEM) to solve heat transfer equations. A brief analysis of the association among Reynolds number with pressure drop and fanning friction factor is included in this paper. Our model can be mounted on building rooftops or open fields where air velocity will be controlled mechanically; thus, it has many applications. This model can be implemented within an agricultural photovoltaic (APV) system, domestic functions, dry agricultural products, and provide heat for greenhouses. The result indicates that 302-514 W thermal energy has been produced for 0.015-0.535 kg/s. For growing inflow temperature, despite the reduction in electrical efficiency, the value of adding electrical and thermal efficiency (overall efficiency) comes with elevation. A 5 °C increase in inflow temperature leads to an overall efficiency increase of 0.33%. This study's findings can help researchers better comprehend air's properties as a heat exchanger in a developed design, and they can be applied to government and commercial projects.

Unsteady magneto porous convective transport by micropolar binary fluid due to inclined plate: An inclusive analogy.

In this investigation , the consequence of viscous dissipation on the unstable magneto porous convective transport by a micropolar binary fluid due to an inclined surface with viscous dissipation and thermal radiation is examined. Viscous dissipation plays a noteworthy role in industrial applications. The governing PDEs are converted to combined ODEs with the Boussinesq approximation using a similarity analysis. The obtained non-linear ODEs are resolved using the shooting method with "ODE45 MATLAB" coding assistance. The numerical outcomes are revealed graphically for various dimensionless parameters and numbers, including temperature, concentration, velocity, and micro-rotation. The temperature, micro-rotation, and velocity fields escalate with increasing Eckert numbers. The radiation parameter and variable viscosity parameter increase the flow rate of the fluid. Increasing radiation parameters, suction parameters, and Prandtl numbers lessen the fluid temperature. The buoyancy parameters have symmetrical impacts on the velocity and microrotation of fluid particles in the cooling and heating modes. Improving Eckert number, inclined angle, Schmidt number, Prandtl number, and magnetic parameter reduces skin friction. The heat transmission rate escalates in quantity due to larger Prandtl number values. Rising Prandtl, Eckert, and Schmidt numbers accelerate the mass transfer rate. The current research result is compared to previously published article's result with good agreement.

Melting and heat management of phase change material using smart ternary-hybrid coolant.

This research investigates the effectiveness of using a smart ternary-hybrid nanofluid to enhance the melting rate and convective behavior of electrically conducting tin (Sn) in a rectangular enclosure under the influence of a uniform magnetic field. The enclosure has adiabatic vertical walls with hot and cold temperatures on the bottom and top walls. The finite element method (FEM) is used to solve the governing equations with appropriate boundary conditions using Galerkin's weighted residual approach. The study focuses on applying tin as the phase change material (PCM), with the highest temperature of 508 K, the lowest temperature of 503 K, and the melting interface temperature of 505 K. To enhance the heat transfer performance, tin-based ternary (graphene (G), silicon carbide (SiC), and nickel (Ni)) hybrid smart coolant is applied into the system. To investigate the mechanism of the melting and convective thermal transfer process, the results of the present study are reported with time for various values of the magnetic field (Ha) and solid concentration of ternary hybrid nanoparticle ( ϕ ). This study represents the streamlines, isothermal lines, melting interface, melting fraction, and heat transfer for the above-mentioned parameters. The results show that increasing the magnetic field reduces the rate of thermal transport by 38.96 % at t = 4000s. However, at a particular time of 2500s, increasing the solid volume fraction of nanoparticles enhances the melting fraction by approximately 8.34 %. Two regression equations are derived for the Nusselt number and melting fraction, with multiple response variables. This article improves understanding of natural convective heat transport during phase change processes for various coolants in different engineering applications.

Ionanofluid flow through a triangular grooved microchannel heat sink: Thermal heightening.

With recent technological advances, thermal transport from different electronic and electrical devices is the most vital concern. The microchannel heat sink (MCHS) of liquid cooling is a useful device to remove over thermal load. Ionanofluid is a brand new and super potential cooling fluid for its ionic conductivity, non-flammability, negligible volatility, and high-level heat stability. In this research, the ionanofluid's velocity and thermal field characteristics through a triangular grooved MCHS are investigated using numerical tools. The combination of ionic liquid (IL) 1-Butyl-3-methylimidazolium Bis(trifluoromethanesulfonyl)imide [C4mim]NTf2 and propylene glycol (PG) is used as base fluid whereas graphene (G) and single-walled carbon nanotube (SWCNT) are chosen as hybrid nanoparticles to make the working ionanofluid. The governing equations of nonlinear partial differential equations describing the physical phenomena along with proper border settings are resolved by applying the finite element method (FEM). Different ratios of hybrid nanoparticles (G: SWCNT) like (1: 0, 1/3: 2/3, 1/2: 1/2, 2/3: 1/3, 0: 1) are suspended in the base fluid mixture. In addition, the base fluid mixture is assumed in different combinations of (IL: PG) as (100: 0, 50: 50, 0: 100). The numerical results are displayed in the forms of streamlines, isothermal lines, and rate of thermal transfer for the pertinent parameters namely forced convection (Re = 100-900) and solid concentration (φ = 0.001-0.05). Also, pressure drop, field synergy number, relative fanning friction feature, relative Nusselt number, and temperature enhancement efficiency are calculated. The results indicate that a higher heat transport rate is found using the IL-based ionanofluid with the highest solid concentration. Moreover, the higher forced convection enhances the thermal efficiency of MCHS. Two linear regression equations along with very good correlation coefficients have been derived from the numerical results.

Axisymmetric boundary layer slip flow with heat transfer over an exponentially stretching bullet-shaped object: A numerical assessment.

The slip flow and thermal transfer inside the boundary layer are extremely significant for various problems in aerodynamics, wing stall, skin friction drag on an entity, high-level velocity aircraft, etc. The current research investigated the effect of the slip factor and shape factor on the axisymmetric bullet-shaped object by taking the viscous dissipation parameter and location parameter. The analysis is conducted for both fixed and moving bullet-shaped objects due to thinner and thicker surfaces. The governing equations are transformed into a system of ordinary differential equations using suitable local axisymmetric similarity transformations and solved by applying the spectral quasi-linearization method. A new correlation analysis is made for velocity and temperature gradients. It is observed that the boundary layer structure has no defined shape due to a thicker bullet-shaped object instead it forms a steep angle with the axis which is contradictory to the formation of the boundary layer. A negative correlation is observed for the parameters M, Ec, Q*, and s but a positive correlation is observed for the parameters such as Pr, P, λ, and ε. The surface thickness and stretching ratio significantly affect the fluid flow and heat transfer processes. It is also noticed that the thinner bullet-shaped object performs as a better cooling conductor compared to a thicker one. The skin friction is reduced in the case of a thinner bullet-shaped object compared to a thicker one. The present analysis reveals that the heat transfer rate and the friction factor may be helpful in industrial sectors for controlling the cooling rate and quality of the final product. This research brings forward to increase in the rate of heat transfer inside the boundary layer region. The result may help to design the various types of moving objects in the automobile engineering sector when the objects pass through the fluid.

Rational optimization of a transcription factor activation domain inhibitor.

Transcription factors are among the most attractive therapeutic targets but are considered largely 'undruggable' in part due to the intrinsically disordered nature of their activation domains. Here we show that the aromatic character of the activation domain of the androgen receptor, a therapeutic target for castration-resistant prostate cancer, is key for its activity as transcription factor, allowing it to translocate to the nucleus and partition into transcriptional condensates upon activation by androgens. On the basis of our understanding of the interactions stabilizing such condensates and of the structure that the domain adopts upon condensation, we optimized the structure of a small-molecule inhibitor previously identified by phenotypic screening. The optimized compounds had more affinity for their target, inhibited androgen-receptor-dependent transcriptional programs, and had an antitumorigenic effect in models of castration-resistant prostate cancer in cells and in vivo. These results suggest that it is possible to rationally optimize, and potentially even to design, small molecules that target the activation domains of oncogenic transcription factors.

Mathematical modeling of unsteady flow with uniform/non-uniform temperature and magnetic intensity in a half-moon shaped domain.

The mathematical modeling of two-dimensional unsteady free convective flow and thermal transport inside a half-moon shaped domain charged in the presence of uniform/non-uniform temperature and magnetic effects with Brownian motion of the nanoparticles has been conducted. Thirty-two types of nanofluids in a combination of various nanoparticles and base fluids having different sizes, shapes, and solid concentrations of nanoparticles are chosen to examine the better performance of heat transfer. The circular boundary is cooled while the diameter boundary is heated with uniform/non-uniform temperature. An external uniform/non-uniform/periodic magnetic field is imposed along diameter. The powerful partial differential equations solver, finite element method of Galerkin type, has been engaged in numerical simulation. The numerical solution's heat transfer mechanism reaches a steady state from the unsteady situation within a very short dimensionless time of about 0.65. The thermal transport rate enhances for increasing buoyancy force whereas decreases with higher magnetic intensity. The uniform thermal condition along the diameter of half-moon gives a higher thermal transport rate compared to non-uniform heating conditions. The non-uniform magnetic field provides greater values of the mean Nusselt number than the uniform field. In addition, the outcomes also predict that a better rate of temperature transport for kerosene-based nanofluid than water-based, ethylene glycol-based, and engine oil-based nanofluid. The heat transfer rate is observed at about 67.86 and 23.78% using Co-Kerosene and Co-water nanofluids, respectively, with an additional 1% nanoparticles volume fraction. The blade shape nanoparticles provide a better heat transfer rate than spherical, cylindrical, brick, and platelet shapes. Small size nanoparticles confirm a higher value of average Nusselt number than big size. Mean Nusselt number increases 22.1 and 5.4% using 1% concentrated Cu-water and Cu-engine oil nanofluid, respectively than base fluid.

Differential Gene Expression Profiles between N-Terminal Domain and Ligand-Binding Domain Inhibitors of Androgen Receptor Reveal Ralaniten Induction of Metallothionein by a Mechanism Dependent on MTF1.

Hormonal therapies for prostate cancer target the androgen receptor (AR) ligand-binding domain (LBD). Clinical development for inhibitors that bind to the N-terminal domain (NTD) of AR has yielded ralaniten and its analogues. Ralaniten acetate is well tolerated in patients at 3600 mgs/day. Clinical trials are ongoing with a second-generation analogue of ralaniten. Binding sites on different AR domains could result in differential effects on AR-regulated gene expression. Here, we provide the first comparison between AR-NTD inhibitors and AR-LBD inhibitors on androgen-regulated gene expression in prostate cancer cells using cDNA arrays, GSEA, and RT-PCR. LBD inhibitors and NTD inhibitors largely overlapped in the profile of androgen-induced genes that they each inhibited. However, androgen also represses gene expression by various mechanisms, many of which involve protein-protein interactions. De-repression of the transcriptome of androgen-repressed genes showed profound variance between these two classes of inhibitors. In addition, these studies revealed a unique and strong induction of expression of the metallothionein family of genes by ralaniten by a mechanism independent of AR and dependent on MTF1, thereby suggesting this may be an off-target. Due to the relatively high doses that may be encountered clinically with AR-NTD inhibitors, identification of off-targets may provide insight into potential adverse events, contraindications, or poor efficacy.

Pin1 inhibition improves the efficacy of ralaniten compounds that bind to the N-terminal domain of androgen receptor.

Therapies for lethal castration-resistant prostate cancer (CRPC) are an unmet medical need. One mechanism underlying CRPC and resistance to hormonal therapies is the expression of constitutively active splice variant(s) of androgen receptor (AR-Vs) that lack its C-terminus ligand-binding domain. Transcriptional activities of AR-Vs and full-length AR reside in its N-terminal domain (NTD). Ralaniten is the only drug proven to bind AR NTD, and it showed promise of efficacy in Phase 1 trials. The peptidyl-prolyl isomerase Pin1 is frequently overexpressed in prostate cancer. Here we show that Pin1 interacted with AR NTD. The inhibition of Pin1 expression or its activity selectively reduced the transcriptional activities of full-length AR and AR-V7. Combination of Pin1 inhibitor with ralaniten promoted cell cycle arrest and had improved antitumor activity against CRPC xenografts in vivo compared to individual monotherapies. These findings support the rationale for therapy that combines a Pin1 inhibitor with ralaniten for treating CRPC.

Ralaniten Sensitizes Enzalutamide-Resistant Prostate Cancer to Ionizing Radiation in Prostate Cancer Cells that Express Androgen Receptor Splice Variants.

Blocking androgen receptor (AR) transcriptional activity by androgen deprivation therapy (ADT) improves the response to radiotherapy for intermediate and high risk prostate cancer. Unfortunately, ADT, antiandrogens, and abiraterone increase expression of constitutively active splice variants of AR (AR-Vs) which regulate DNA damage repair leading to resistance to radiotherapy. Here we investigate whether blocking the transcriptional activities of full-length AR and AR-Vs with ralaniten leads to enhanced sensitivity to radiotherapy. Combination therapies using ralaniten with ionizing radiation were evaluated for effects on proliferation, colony formation, cell cycle, DNA damage, and Western blot analyses in human prostate cancer cells that express both full-length AR and AR-Vs. Ralaniten and a potent next-generation analog (EPI-7170) decreased expression of DNA repair genes whereas enzalutamide had no effect. FACS analysis revealed a dose-dependent decrease of BrdU incorporation with increased accumulation of γH2AX with a combination of ionizing radiation with ralaniten. An additive inhibitory effect on proliferation of enzalutamide-resistant cells was achieved with a combination of ralaniten compounds with ionizing radiation. Ralaniten and EPI-7170 sensitized prostate cancer cells that express full-length AR and AR-Vs to radiotherapy whereas enzalutamide had no added benefit.

Inhibition of androgen receptor by decoy molecules delays progression to castration-recurrent prostate cancer.

Androgen receptor (AR) is a member of the steroid receptor family and a therapeutic target for all stages of prostate cancer. AR is activated by ligand binding within its C-terminus ligand-binding domain (LBD). Here we show that overexpression of the AR NTD to generate decoy molecules inhibited both the growth and progression of prostate cancer in castrated hosts. Specifically, it was shown that lentivirus delivery of decoys delayed hormonal progression in castrated hosts as indicated by increased doubling time of tumor volume, prolonged time to achieve pre-castrate levels of serum prostate-specific antigen (PSA) and PSA nadir. These clinical parameters are indicative of delayed hormonal progression and improved therapeutic response and prognosis. Decoys reduced the expression of androgen-regulated genes that correlated with reduced in situ interaction of the AR with androgen response elements. Decoys did not reduce levels of AR protein or prevent nuclear localization of the AR. Nor did decoys interact directly with the AR. Thus decoys did not inhibit AR transactivation by a dominant negative mechanism. This work provides evidence that the AR NTD plays an important role in the hormonal progression of prostate cancer and supports the development of AR antagonists that target the AR NTD.

Targeting Androgen Receptor Activation Function-1 with EPI to Overcome Resistance Mechanisms in Castration-Resistant Prostate Cancer.

PURPOSE: Persistent androgen receptor (AR) transcriptional activity is clinically evident in castration-resistant prostate cancer (CRPC). Therefore, AR remains as a viable therapeutic target for CRPC. All current hormonal therapies target the C-terminus ligand-binding domain (LBD) of AR. By using EPI to target AR activation function-1 (AF-1), in the N-terminal domain that is essential for AR transactivation, we evaluate the ability of EPI to overcome several clinically relevant AR-related mechanisms of resistance. EXPERIMENTAL DESIGN: To study the effect of EPI on AR transcriptional activity against overexpressed coactivators, such as SRC1-3 and p300, luciferase reporter assays were performed using LNCaP cells. AR-negative COS-1 cells were employed for reporter assays to examine whether the length of polyglutamine tract affects inhibition by EPI. The effect of EPI on constitutively active AR splice variants was studied in LNCaP95 cells, which express AR-V7 variant. To evaluate the effect of EPI on the proliferation of LNCaP95 cells, we performed in vitro BrdUrd incorporation assay and in vivo studies using xenografts in mice. RESULTS: EPI effectively overcame several molecular alterations underlying aberrant AR activity, including overexpressed coactivators, AR gain-of-function mutations, and constitutively active AR-V7. EPI inhibited AR transcriptional activity regardless of the length of polyglutamine tract. Importantly, EPI significantly inhibited the in vitro and in vivo proliferation of LNCaP95 prostate cancer cells, which are androgen independent and enzalutamide resistant. CONCLUSIONS: These findings support EPI as a promising therapeutic agent to treat CRPC, particularly against tumors driven by constitutively active AR splice variants that are resistant to LBD-targeting drugs. Clin Cancer Res; 22(17); 4466-77. ©2016 AACRSee related commentary by Sharp et al., p. 4280.

Cotargeting Androgen Receptor Splice Variants and mTOR Signaling Pathway for the Treatment of Castration-Resistant Prostate Cancer.

PURPOSE: The PI3K/Akt/mTOR pathway is activated in most castration-resistant prostate cancers (CRPC). Transcriptionally active androgen receptor (AR) plays a role in the majority of CRPCs. Therefore, cotargeting full-length (FL) AR and PI3K/Akt/mTOR signaling has been proposed as a possible, more effective therapeutic approach for CRPC. However, truncated AR-splice variants (AR-V) that are constitutively active and dominant over FL-AR are associated with tumor progression and resistance mechanisms in CRPC. It is currently unknown how blocking the PI3K/Akt/mTOR pathway impacts prostate cancer driven by AR-Vs. Here, we evaluated the efficacy and mechanism of combination therapy to block mTOR activity together with EPI-002, an AR N-terminal domain (NTD) antagonist that blocks the transcriptional activities of FL-AR and AR-Vs in models of CRPC. EXPERIMENTAL DESIGN: To determine the functional roles of FL-AR, AR-Vs, and PI3K/Akt/mTOR pathways, we employed EPI-002 or enzalutamide and BEZ235 (low dose) or everolimus in human prostate cancer cells that express FL-AR or FL-AR and AR-Vs (LNCaP95). Gene expression and efficacy were examined in vitro and in vivo RESULTS: EPI-002 had antitumor activity in enzalutamide-resistant LNCaP95 cells that was associated with decreased expression of AR-V target genes (e.g., UBE2C). Inhibition of mTOR provided additional blockade of UBE2C expression. A combination of EPI-002 and BEZ235 decreased the growth of LNCaP95 cells in vitro and in vivo CONCLUSIONS: Cotargeting mTOR and AR-NTD to block transcriptional activities of FL-AR and AR-Vs provided maximum antitumor efficacy in PTEN-null, enzalutamide-resistant CRPC. Clin Cancer Res; 22(11); 2744-54. ©2015 AACR.

An imaging agent to detect androgen receptor and its active splice variants in prostate cancer.

Constitutively active splice variants of androgen receptor (AR-Vs) lacking ligand-binding domain (LBD) are a mechanism of resistance to androgen receptor LBD-targeted (AR LBD-targeted) therapies for metastatic castration-resistant prostate cancer (CRPC). There is a strong unmet clinical need to identify prostate cancer patients with AR-V-positive lesions to determine whether they will benefit from further AR LBD-targeting therapies or should receive taxanes or investigational drugs like EPI-506 or galeterone. Both EPI-506 (NCT02606123) and galeterone (NCT02438007) are in clinical trials and are proposed to have efficacy against lesions that are positive for AR-Vs. AR activation function-1 (AF-1) is common to the N-terminal domains of full-length AR and AR-Vs. Here, we provide proof of concept for developing imaging compounds that directly bind AR AF-1 to detect both AR-Vs and full-length AR. 123I-EPI-002 had specific binding to AR AF-1, which enabled direct visualization of CRPC xenografts that express full-length AR and AR-Vs. Our findings highlight the potential of 123I-EPI-002 as an imaging agent for the detection of full-length AR and AR-Vs in CRPC.

Osteoblast-derived factors induce androgen-independent proliferation and expression of prostate-specific antigen in human prostate cancer cells.

PURPOSE: Prostate cancer metastasizes to the skeleton to form osteoblastic lesions. Androgen ablation is the current treatment for metastatic prostate cancer. This therapy is palliative, and the disease will return in an androgen-independent form that is preceded by a rising titer of prostate-specific antigen (PSA). Here, we investigated the possibility that human osteoblasts might secrete factors that contribute to the emergence of androgen-independent prostate cancer. EXPERIMENTAL DESIGN: Primary cultures of human osteoblasts were used as a source of conditioned medium (OCM). Proliferation, expression of androgen-regulated genes, and transactivation of the androgen receptor (AR) were monitored in LNCaP human prostate cancer cells in response to OCM using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, Northern blot analysis, and reporter gene constructs. Levels of interleukin-6 (IL-6) present in OCM were measured, and its contribution to proliferation and expression of PSA were investigated by neutralization studies with anti IL-6 antibodies. RESULTS: OCM increased the proliferation and expression of PSA at both the protein and RNA levels in LNCaP cells. Synergistic increases in the activities of PSA (6.1 kb)- and pARR(3)-tk-luciferase reporters were measured in cells cotreated with both OCM and androgen. OCM targeted the NH(2)-terminal domain of the AR. The effect of OCM on transcriptional activity of the AR was inhibited by an antiandrogen. Neutralizing antibodies to IL-6 blocked proliferation and expression of PSA by OCM. CONCLUSION: Osteoblasts secrete factors, such as IL-6, that cause androgen-independent induction of PSA gene expression and proliferation of prostate cancer cells by a mechanism that partially relies on the AR. Identifying such molecular mechanisms may lead to improved clinical management of metastatic prostate cancer.

Characterization of niphatenones that inhibit androgen receptor N-terminal domain.

Androgen ablation therapy causes a temporary reduction in tumor burden in patients with advanced prostate cancer. Unfortunately the malignancy will return to form lethal castration-recurrent prostate cancer (CRPC). The androgen receptor (AR) remains transcriptionally active in CRPC in spite of castrate levels of androgens in the blood. AR transcriptional activity resides in its N-terminal domain (NTD). Possible mechanisms of continued AR transcriptional activity may include, at least in part, expression of constitutively active splice variants of AR that lack the C-terminal ligand-binding domain (LBD). Current therapies that target the AR LBD, would not be effective against these AR variants. Currently no drugs are clinically available that target the AR NTD which should be effective against these AR variants as well as full-length AR. Niphatenones were originally isolated and identified in active extracts from Niphates digitalis marine sponge. Here we begin to characterize the mechanism of niphatenones in blocking AR transcriptional activity. Both enantiomers had similar IC50 values of 6 µM for inhibiting the full-length AR in a functional transcriptional assay. However, (S)-niphatenone had significantly better activity against the AR NTD compared to (R)-niphatenone. Consistent with niphatenones binding to and inhibiting transactivation of AR NTD, niphatenones inhibited AR splice variant. Niphatenone did not affect the transcriptional activity of the related progesterone receptor, but slightly decreased glucocorticoid receptor (GR) activity and covalently bound to GR activation function-1 (AF-1) region. Niphatenone blocked N/C interactions of AR without altering either AR protein levels or its intracellular localization in response to androgen. Alkylation with glutathione suggests that niphatenones are not a feasible scaffold for further drug development.

Spongian diterpenoids inhibit androgen receptor activity.

Androgen receptor is a ligand-activated transcription factor and a validated drug target for all stages of prostate cancer. Antiandrogens compete with physiologic ligands for androgen receptor ligand-binding domain (LBD). High-throughput screening of a marine natural product library for small molecules that inhibit androgen receptor transcriptional activity yielded the furanoditerpenoid spongia-13(16),-14-dien-19-oic acid, designated terpene 1 (T1). Characterization of T1 and the structurally related semisynthetic analogues (T2 and T3) revealed that these diterpenoids have antiandrogen properties that include inhibition of both androgen-dependent proliferation and androgen receptor transcriptional activity by a mechanism that involved competing with androgen for androgen receptor LBD and blocking essential N/C interactions required for androgen-induced androgen receptor transcriptional activity. Structure-activity relationship analyses revealed some chemical features of T1 that are associated with activity and yielded T3 as the most potent analogue. In vivo, T3 significantly reduced the weight of seminal vesicles, which are an androgen-dependent tissue, thereby confirming the on-target activity of T3. The ability to create analogues of diterpenoids that have varying antiandrogen activity represents a novel class of chemical compounds for the analysis of androgen receptor ligand-binding properties and therapeutic development.