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Becoming a mother is often accompanied by a loss of social connections, which can reduce the availability of social support. This can increase maternal stress with negative health outcomes. Therefore, we examined how mothers' social contact with other mothers living in the same neighborhood can form a compensative source of social support and wellbeing. Data was collected from mothers (N = 443) of a child under school age while visiting the public maternity and child health clinics located in two neighborhoods in Helsinki, Finland. We found that mothers' frequent and positive contact with other local mothers was positively related with their life satisfaction through identification-based social support. Contact also had a specific indirect effect on life satisfaction through social support. Frequent positive contact with other local mothers can serve to scaffold the emergence of social identification, which in turn unlocks further social support with positive health effects.

Binding of steroid hormones to their cognate receptors regulates the growth of most prostate and breast cancers. We hypothesized that CYP11A inhibition might halt the synthesis of all steroid hormones, because CYP11A is the only enzyme that catalyses the first step of steroid hormone biosynthesis. We speculated that a CYP11A inhibitor could be administered safely provided that the steroids essential for life are replaced. Virtual screening and systematic structure–activity relationship optimization were used to develop ODM-208, the first-in-class, selective, nonsteroidal, oral CYP11A1 inhibitor. Safety of ODM-208 was assessed in rats and Beagle dogs, and efficacy in a VCaP castration-resistant prostate cancer (CRPC) xenograft mouse model, in mice and dogs, and in six patients with metastatic CRPC. Blood steroid hormone concentrations were measured using liquid chromatography-mass spectrometry. ODM-208 binds to CYP11A1 and inhibited its enzymatic activity. ODM-208 administration led to rapid, complete, durable, and reversible inhibition of the steroid hormone biosynthesis in an adrenocortical carcinoma cell model in vitro, in adult noncastrated male mice and dogs, and in patients with CRPC. All measured serum steroid hormone concentrations reached undetectable levels within a few weeks from the start of ODM-208 administration. ODM-208 was well tolerated with steroid hormone replacement. The toxicity findings were considered related to CYP11A1 inhibition and were reversed after stopping of the compound administration. Steroid hormone biosynthesis can be effectively inhibited with a small-molecule inhibitor of CYP11A1. The findings suggest that administration of ODM-208 is feasible with concomitant corticosteroid replacement therapy.

Supplementary Data from First-in-Class Small Molecule to Inhibit CYP11A1 and Steroid Hormone Biosynthesis

5057 Background: Activating AR mutations ensure a continued AR activation by non-androgen steroid ligands, e.g. progesterone and glucocorticoids. CYP11A1 is the only enzyme that catalyzes the conversion of cholesterol to pregnenolone, from which all steroid hormones (glucocorticoids, mineralocorticoids, and sex steroids) are subsequently derived. ODM-208, an oral, selective inhibitor of CYP11A1, is being evaluated for safety and efficacy as a treatment of mCRPC in the ongoing CYPIDES phase I/II trial in men previously treated with both novel hormonal therapies and taxanes (ClinicalTrials.gov identifier: NCT03436485). Preliminary phase 1 results were previously reported (Fizazi K et al., ASCO GU 2022). Here we confirm that the in-vitro sensitivity of common AR mutations to ODM-208 treatment is mirrored in patient response in CYPIDES phase 1. Methods: ODM-208 was administered at daily doses between 6-150 mg (phase 2 dose: 10 mg) with dexamethasone and fludrocortisone, resulting in maximal suppression in all measured steroids at all doses. AR ligand-binding domain (LBD) mutations (L702H, V716M, W742L, W742C, H875Y, F877L, T878A, T878S, M896T, M896V) were assessed using a BEAMing assay (Sysmex Inostics) from plasma circulating cell-free DNA (cfDNA) collected before the first dose of ODM-208. The activation of wild-type (wt) and LBD mutated AR with various ligands was also studied in vitro using a luciferase reporter assay in AR negative PC3 cells. Results: 17 of 44 patients had at least one AR activating mutation, the most frequent being L702H (n = 11), T878A (n = 10), and H875Y (n = 6). Eleven out of the 16 evaluable patients (68%) with an AR LBD mutation achieved ≥50% reduction in serum PSA compared with 2 out of the 24 evaluable patients (8%) without an AR LBD mutation (P

Social media is increasingly becoming a forum for criminality, misuse, and hate speech, as there are no filters or other controlling mechanisms to filter user-generated content.Furthermore, disinformation and propaganda are becoming more sophisticated and harder to track. Hence, this dark side of social media can pose a viable threat to national security. Future generations will be born into an environment of polluted and polarised online information networks. Consequently, young people, many of whom use social media on a daily basis, will have to find ways to survive in these circumstances,often without the help, knowledge, or experience of earlier generations. Thus, young people are at risk of becoming predisposed to all kinds of harmful material, which, in turn, can affect their thinking and behaviour. This can lead to many new threats to national security. This study focuses on the observations of police officers on the current trends and threats youngsters face on the dark side of social media. The aim was to examine possible threats to national security related to young people’s social media use based on data from three semi-structured interviews with police officers working in Preventive Measures Units. The analysis was done using inductive content analysis. Based on the analysis, there are three main threats to national security concerning young people’s social media use: the amount of false information available online, the glorification of violence and crime, and large-scale gatherings and swarming. The results indicate that although most young people’s social media use is harmless, social media platforms can also be used in a way that threatens national security. Many of the threats posed by young people’s social media use have not yet been realised. However, it is important to be aware of the risks and be prepared for any possible negative outcomes in order to maintain national security. peerReviewed

Background: Prostate cancer is a major global challenge due to the increasing number of aging population and frequency of diagnosis. During the past decade new treatments have been targeted to the androgen signaling axis either by inhibiting the binding of androgens to androgen receptor (AR) and AR nuclear translocation, or by inhibiting androgen production via CYP17A1 enzyme. Despite the significant progress on the research and new therapies, CRPC is still incurable and there is urgent need for better, more effective treatments. ODM-208 is an oral, non-steroidal and selective inhibitor of CYP11A1 enzyme, suppressing the synthesis of all steroid hormones that could be potential agonistic AR ligands. Methods: The inhibition of CYP11A1 was measured in vitro by the formation of radiolabelled isocapronic acid in a human adrenal cortex cell line (H295R). The tumor growth inhibition of ODM-208 was studied in VCaP castration-resistant prostate cancer (CRPC) xenograft model. At the end of the xenograft study, plasma ACTH and LH, and key steroid hormone concentrations were analysed from plasma and target tissues. In addition, full length AR (AR-FL) and AR-V7 were analysed from the tumors by western blot and key enzymes of androgen biosynthesis, CYP17A1, AKR1C3, SRD5A1 were quantified by qPCR. In dogs an ACTH stimulation test was done. Results: ODM-208 potently inhibits CYP11A1 enzyme in vitro. In addition, in vivo in the VCaP CRPC xenograft model ODM-208 significantly inhibited tumor growth. Importantly, the amount of AR-FL and AR-V7 levels remained unchanged in the tumors after ODM-208 treatment. Slight increase of CYP17A1 and SRD5A1 enzyme levels was observed, indicating the activation of steroidogenesis in VCaP tumors in vivo. Treatment had no effect on plasma LH, whereas ACTH was significantly increased demonstrating reduction in glucocorticoid levels by negative feedback. In line with ACTH, all measured steroid hormones were significantly reduced both in plasma and target tissues. In dogs ACTH-stimulated cortisol production was significantly inhibited after single oral dose of ODM-208. Conclusions: ODM-208 shows promising antitumor activity in preclinical CRPC models and has favorable toxicological profile. Thus, ODM-208 might have potential for treating patients with CRPC. Based on the nonclinical results, a phase 1/2 clinical trial has been initiated. Unless otherwise noted, all abstracts presented at ENDO are embargoed until the date and time of presentation. For oral presentations, the abstracts are embargoed until the session begins. Abstracts presented at a news conference are embargoed until the date and time of the news conference. The Endocrine Society reserves the right to lift the embargo on specific abstracts that are selected for promotion prior to or during ENDO.

We report the discovery of a novel nonsteroidal dual-action compound, ODM-204, that holds promise for treating patients with castration-resistant prostate cancer (CRPC), an advanced form of prostate cancer characterised by high androgen receptor (AR) expression and persistent activation of the AR signaling axis by residual tissue androgens. For ODM-204, has a dual mechanism of action. The compound is anticipated to efficiently dampen androgenic stimuli in the body by inhibiting CYP17A1, the prerequisite enzyme for the formation of dihydrotestosterone (DHT) and testosterone (T), and by blocking AR with high affinity and specificity. In our study, ODM-204 inhibited the proliferation of androgen-dependent VCaP and LNCaP cells in vitro and reduced significantly tumour growth in a murine VCaP xenograft model in vivo. Intriguingly, after a single oral dose of 10-30 mg/kg, ODM-204 dose-dependently inhibited adrenal and testicular steroid production in sexually mature male cynomolgus monkeys. Similar results were obtained in human chorionic gonadotropin-treated male rats. In rats, leuprolide acetate-mediated (LHRH agonist) suppression of the circulating testosterone levels and decrease in weights of androgen-sensitive organs was significantly and dose-dependently potentiated by the co-administration of ODM-204. ODM-204 was well tolerated in both rodents and primates. Based on our data, ODM-204 could provide an effective therapeutic option for men with CRPC.

Introduction: The bromodomain and extraterminal (BET) family of proteins are chromatin readers that promote the transcription of several important cell identity genes. BET proteins also control expression of many genes that play an essential role in the pathogenesis of human cancer, including cell-cycle and proliferation-regulating genes. The small-molecule BET inhibitors block BET binding to chromatin and have shown antitumor activity in a variety of pre-clinical cancer models. In this study, we evaluated the anticancer activity of the novel BET inhibitor, ODM-207, in ER+ breast cancer models as a single agent and in combination with other cancer drugs. Methods: ER+ breast cancer cell lines were studied for sensitivity to ODM-207 and the in vivo efficacy was assessed using the ER+ Ma3366 patient-derived xenograft model. For gene expression analyses, breast cancer cells were treated with ODM-207 or reference BET inhibitor JQ1 and differentially expressed genes were analyzed by RNA-sequencing. The ability of ODM-207 to regulate anticancer signaling pathways was validated by western blotting. Synergistic drug interactions were profiled using five-concentration dose response matrices. Results: ODM-207 is a novel BET inhibitor structurally distinct from JQ1 that shows antiproliferative activity in a broad panel of cancer cell lines. The strongest antitumor activity could be observed in hormone-dependent prostate and breast cancer models. In this study, we show that ODM-207 effectively inhibits the proliferation of ER+ breast cancer cell lines by inducing cell cycle arrest in G0/G1-phase. Additionally, ODM-207 suppresses the growth of ER+ patient-derived breast cancer xenograft tumors. ODM-207 as well as JQ1 targeted several pathways important for cancer progression such as MYC, estrogen response and cell cycle gene signatures. The inhibition of key cell cycle regulators, such as CDK4 and Cyclin D1, were further verified. The cyclin D1:CDK4/6 axis plays a significant role in the development, and currently, treatment of ER+ breast cancer together with endocrine therapy. Interestingly, ODM-207 was shown to synergize with palbociclib in vitro in ER+ breast cancer cell lines: the combination of ODM-207 and CDK4/6 inhibitor palbociclib achieved greater cell proliferation inhibition than either drug alone at sub IC50 concentrations. Notably, the ODM-207 and palbociclib combination did not cause the induction of an obvious senescent-like phenotype as compared to palbociclib alone, but rather affected cell survival cellular assays. Conclusions: In summary, ODM-207, which is currently in Phase I clinical trials for treating solid tumors, causes significant growth inhibition in pre-clinical models of ER+ breast cancer and enhances antiproliferative activity of palbociclib, providing a rationale for development of a combination therapy. Citation Format: Julia Lindqvist, Mari Björkman, Reetta Riikonen, Daniel Nicorici, Elina Mattila, Mahaboobi Jaleel, Chandrasekhar Abbineni, Anu-Maarit Moilanen. Antitumor activity of ODM-207, a novel BET bromodomain inhibitor, in nonclinical models of ER+ breast cancer as single agent and as a combination treatment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3827.

340 Background: Castration-resistant prostate cancer (CRPC) is a major cause of cancer mortality worldwide. The mechanisms behind the development of resistance are complex and not fully understood; altered androgen synthesis, androgen receptor (AR) overexpression or gene amplification, and mutations have been indentified. However, tumor growth may still be responsive to therapies that can further suppress de novo intratumoral steroid synthesis upstream of CYP17A1. ODM-208 is an oral, non-steroidal and selective inhibitor of CYP11A1 enzyme that suppresses the synthesis of all steroid hormones and their precursors. Methods: The inhibition of CYP11A1 was measured in vitro by the formation of radiolabelled isocapronic acid in a human adrenal cortex cell line (H295R), and further analysing pregnenolone (Preg) and testosterone (T) formation by ELISA. The tumor growth inhibition of ODM-208 alone or in combination with prednisone (Pred) was studied in VCaP CRPC xenograft where also concentrations of main steroid hormones progesterone (P), corticosterone (CORT) and T in tumors and adrenals were analysed. In addition, plasma ACTH and LH levels were measured at the end of the xenograft study. In dogs an ACTH stimulation test was done. Toxicity studies were conducted in rats and dogs. Results: ODM-208 potently inhibits CYP11A1 and synthesis of Preg and T with nM concentrations in vitro. In the VCaP CRPC xenograft ODM-208 alone and in combination with Pred significantly inhibited tumor growth. Concentrations of T, P and CORT were significantly decreased in the adrenals, indicating strong CYP11A1 inhibition. Also, significantly decreased steroid levels in tumors was observedhe Pred combination increased plasma ACTH levels less than ODM-208 alone, whereas no difference was seen in the LH. In dogs ACTH-stimulated cortisol production was significantly inhibited after single oral dose of ODM-208. In toxicological studies ODM-208 showed expected reversible findings in target tissues, mainly related to the pharmacology. Conclusions: ODM-208 shows promising antitumor activity in preclinical CRPC models with favorable toxicological profile. Thus, ODM-208 might have potential for treating the patients with CRPC.

Introduction: The bromodomain and extraterminal (BET) family of proteins are chromatin readers that recognize and bind to specific acetylated histones and promote the transcription of several important cell identity genes. BET bromodomain inhibitors have shown promising antitumor activity in a variety of pre-clinical cancer models, as BET inhibition abrogates the transcription of several key oncogenes in a cell type-specific manner. It is known that inhibition of BET proteins effectively inhibits the proliferation of estrogen receptor positive (ER+) breast cancer cells, at least in part through repression of ER and MYC signaling. However, many additional cancer-associated genes are likely to underlie the growth inhibitory effects of BET inhibitors in breast cancer. The purpose of this study was to determine the anticancer activity of the novel BET bromodomain inhibitor ODM-207 in pre-clinical ER+ breast cancer models, and further, to look for cancer-associated signaling pathways suppressed by BET inhibitors. Methodology and results: ODM-207 is a novel, highly selective BET bromodomain inhibitor structurally distinct from JQ1 and its benzodiazepine-related derivatives. In this study, we show that ODM-207 effectively inhibits the proliferation of ER+ breast cancer cell lines when measured by cell viability assays as well as suppresses the growth of patient-derived xenograft tumors. Furthermore, we wanted to investigate the anticancer signaling pathways regulated by ODM-207 as well as the prototypical BET inhibitor JQ1 in breast cancer cells. For this purpose, we performed RNA sequencing on two ER+ breast cancer cell lines after 24h treatment with the aforementioned BET inhibitors. We found that both BET inhibitors targeted several genes and pathways important for breast cancer progression. For example, the targets included CDK4 and CDK6, two cell cycle kinases fundamental for the development and treatment of ER+ breast cancer. The RNA sequencing results were further validated in vitro, and were utilized as a basis for combination therapy assessment. Conclusions: Our results indicate that the novel BET bromodomain inhibitor ODM-207, which is currently in Phase I clinical trials for treating solid tumors, causes significant growth inhibition and cell cycle arrest in pre-clinical models of ER+ breast cancer, and regulates multiple crucial signaling pathways involved in breast cancer cell cycle and survival. Citation Format: Julia Lindqvist, Mari Björkman, Reetta Riikonen, Daniel Nicorici, Elina Mattila, Chandrasekhar Abbineni, Mahaboobi Jaleel, John Eriksson, Pekka Kallio, Anu-Maarit Moilanen. Therapeutic targeting of estrogen receptor positive breast cancer with the BET bromodomain inhibitor ODM-207 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3970.

Article, Activation of androgen receptor (AR) is crucial for prostate cancer growth. Remarkably, also castration-resistant prostate cancer (CRPC) is dependent on functional AR, and several mechanisms have been proposed to explain the addiction. Known causes of CRPC include gene amplification and overexpression as well as point mutations of AR. We report here the pharmacological profile of ODM-201, a novel AR inhibitor that showed significant antitumor activity and a favorable safety profile in phase 1/2 studies in men with CRPC. ODM-201 is a full and high-affinity AR antagonist that, similar to second-generation antiandrogens enzalutamide and ARN-509, inhibits testosterone-induced nuclear translocation of AR. Importantly, ODM-201 also blocks the activity of the tested mutant ARs arising in response to antiandrogen therapies, including the F876L mutation that confers resistance to enzalutamide and ARN-509. In addition, ODM-201 reduces the growth of AR-overexpressing VCaP prostate cancer cells both in vitro and in a castration-resistant VCaP xenograft model. In contrast to other antiandrogens, ODM-201 shows negligible brain penetrance and does not increase serum testosterone levels in mice. In conclusion, ODM-201 is a potent AR inhibitor that overcomes resistance to AR-targeted therapies by antagonizing both overexpressed and mutated ARs. ODM-201 is currently in a phase 3 trial in CRPC., published version, http://purl.org/eprint/status/PeerReviewed

Background: Bromodomain and extra-terminal (BET) family proteins are dual bromodomain-containing epigenetic readers that bind to acetylated-lysine residues at gene promoter and enhancer elements in histones and recruit protein complexes to promote transcriptional elongation. Recent evidence demonstrates that BET bromodomain inhibition leads to anti-proliferative activity in pre-clinical models of many hematological malignancies and solid tumors. Selective inhibition of BET bromodomains by small molecule inhibitors has emerged as a promising therapeutic strategy for the treatment of cancer. In this study, we evaluated the antitumor activity of ODM-207, a novel, potent and highly selective BET bromodomain inhibitor. Methods and Results: ODM-207 is a potent and selective BET inhibitor that is structurally unrelated to the benzodiazepine like inhibitors such as JQ1, I-BET762, and OTX015. We tested the preclinical activity of ODM-207 across multiple tumor types in a 4-day growth inhibition in vitro assay. ODM-207 potently inhibits cell viability of a wide range of hematological and solid tumor cell lines. ODM-207 also shows potent antiproliferative effects in patient-derived cancer cells representing various tumor types. In VCaP prostate cancer cell lines, ODM-207 induced apoptosis consistent with increased expression of pro-apoptotic regulators, whereas potent antiproliferative effects associated with cell cycle arrest and cellular senescence were seen in e.g. LNCaP prostate cancer cell line. To gain insight on mechanisms of acquired BET inhibitor resistance, we generated a cell line with resistance to BET-inhibition by culturing LNCaP prostate cancer cells with increasing concentrations of OTX015, a potent and selective BET-inhibitor. Interestingly, ODM-207 is also able to inhibit proliferation and downregulate Myc levels in cells that have acquired resistance to BET-inhibitors OTX015 and I-BET762. In xenograft models, oral treatment with ODM-207 significantly inhibits tumor growth at a dose which is well tolerated. Conclusions: In summary, these studies demonstrate that ODM-207 is a potent inhibitor of BET proteins in models of hematologic malignancies and solid tumors in vitro and in vivo and support its clinical development for the treatment of cancer. Citation Format: Anu-Maarit Moilanen, Mari Björkman, Reetta Riikonen, Chandrasekhar Abbineni, Mahaboobi Jaleel, Sivapriya Marappan, Tarja Ikonen, Girish Daginakatte, Aravind A B, Elina Mattila, Juha Rantala, Susanta Samajdar, Murali Ramachandra, Pekka Kallio. Targeting cancer with a novel BET bromodomain inhibitor ODM-207 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 118. doi:10.1158/1538-7445.AM2017-118

Introduction: BET (bromodomain and extraterminal) family proteins (BRD2, BRD3, BRD4, and BRDT) are epigenetic readers that bind to acetylated-lysine residues in histones and recruit protein complexes to promote transcription elongation. In many cancers, BET proteins have been shown to regulate expression of MYC and other oncogenic drivers that are important for cell proliferation and survival. Pharmacologic inhibition of the BET-histone interaction has been shown to result in transcriptional downregulation of a number of oncogenes and inhibition of tumor growth providing a novel strategy for treatment of cancer. Therefore, in this study, we evaluated the in vitro and in vivo antitumor activity of ODM-207, a novel, potent and highly selective BET bromodomain inhibitor using cell lines derived from prostate and breast cancer as well as patient-derived tumor cell cultures of breast cancer. Methods and Results: ODM-207 has antiproliferative effects on several hematological and solid tumor cell lines. In a panel of prostate cancer cell lines, ODM-207 attenuates cell growth of androgen receptor (AR)-positive cell lines such as VCaP and 22Rv1. RNA-sequencing and Western blot studies revealed that the exposure of sensitive prostate cancer cells to ODM-207 is associated with rapid down-regulation c-Myc expression levels while wtAR was not affected. In 22Rv1 prostate cancer xenograft, which expresses both the full-length androgen receptor and androgen receptor splice variant V7, oral administration of ODM-207 was very efficacious in suppressing tumor growth at well tolerated doses whereas enzalutamide had only a modest effect. Contrary to our findings in prostate cancer cells, BET-inhibitor treatment of estrogen-dependent MCF-7 breast cancer cell line inhibits tumor growth in cell proliferation assays but is associated with down-regulation of ERα while the c-Myc levels are very low, highlighting the context-dependent functional effects of BET inhibition. Interestingly, ODM-207 produces potent antiproliferative effects associated with cell-cycle arrest and cellular senescence in patient-derived breast cancer cells. Conclusions: In summary, ODM-207 is a new generation BET inhibitor found to possess excellent pharmacological properties and antitumor activity both in vitro and in vivo. Our data suggest the potential utilization of ODM-207 for the treatment of prostate and breast cancer. Citation Format: Mari Björkman, Elina Mattila, Reetta Riikonen, Chandra Sekhar, Mahaboobi Jaleel, Sivapriya Marappan, Tarja Ikonen, Daniel Nicorici Nicorici, Juha Rantala, Susanta Samajdar, Murali Ramachandra, Pekka Kallio, Anu-Maarit Moilanen. ODM-207, a novel BET-bromodomain inhibitor as a therapeutic approach for the treatment of prostate and breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4649.

230 Background: Castration-resistant prostate cancer (CRPC) is characterized by high androgen receptor (AR) expression and persistent activation of AR signaling axis by residual tissue androgens. Inhibiting AR and androgen biosynthesis together may be more effective than inhibiting either alone to treat CRPC. ODM-204 is a potent, orally administered investigational non-steroidal dual inhibitor of CYP17A1 and AR. In vivo, ODM-204 shows favourable pharmacokinetic/ pharmacodynamics (PK/PD) properties in intact mature male monkeys and strong antitumor activity in VCaP xenograft. Methods: The inhibition of CYP17A1 by ODM-204 was studied by using human testicular microsomes and adrenal cortex cell line. Potency to human AR was demonstrated in cells stably transfected with the full-length AR and androgen-responsive reporter gene constructs. PK/PD relationships were evaluated in intact male mature monkeys after single and multiple doses of ODM-204 (10-30 mg/kg/day) by measuring plasma PK after several time points and serum testosterone (T), dehydroepiandrosterone (DHEA), luteinizing hormone (LH), aldosterone, and cortisol 5 hours after the daily dose. Efficacy was further studied in VCaP xenograft in intact male nude mice treated with ODM-204, enzalutamide (enza), abiraterone (abi), or enza+abi-combination for 28 days. Results: ODM-204 binds to AR with a high affinity (Ki47 nM) and selectivity, and has a high potency towards CYP17A1 (IC5022 nM). In intact monkeys, ODM-204 was well tolerated and showed good oral bioavailability with linear and dose proportional pharmacokinetics. Already after a single oral dose of ODM-204, serum T and DHEA were markedly suppressed. After multiple days of dosing, LH was increased, however, changes in cortisol and aldosterone were only minimal. In VCaP xenograft model, ODM-204 showed significant antitumor activity, better than single agent enza or abi, or their combination. Conclusions: ODM-204 is a novel, dual CYP17A1 and AR inhibitor for the treatment of CRPC. A clinical phase 1/2 dose-escalation trial (NCT02344017) is ongoing in patients with mCRPC to determine PK, PD, safety and anti-tumor activity of ODM-204.

221 Background: Castration-resistant prostate cancer (CRPC) is characterized by high androgen receptor (AR) expression and persistent activation of AR signaling axis by residual tissue/tumor androgens. Targeting AR and androgen biosynthesis together may be more effective than either alone. ODM-204 is a novel, non-steroidal dual inhibitor of CYP17A1 and AR, which has shown promising results in preclinical studies. Methods: The binding affinity of ODM-204 to wild type AR was determined in rat prostate cytosolic lysates. The potency and functional activity of ODM-204 to human AR were demonstrated in cells stably transfected with the full-length AR and androgen-responsive reporter gene constructs. In addition, assays for AR nuclear translocation and the transactivation of human AR mutants T877A, W741L, and F876L were conducted. The effects of ODM-204 on the growth of androgen-dependent VCaP and LNCaP cells in vitro and subcutaneously grafted VCaP cells in vivo with the oral dose of 50 mg/kg/day were studied. The inhibition of CYP17A1 by ODM-204 was studied in vitro by using human and rat testicular microsomes and a human adrenal cortex cell line, and in vivo in male rats coadministered with luteinizing hormone releasing hormone agonist leuprolide acetate to mimic clinical situation. Results: ODM-204 is a potent inhibitor of both AR and CYP17A1. It binds to AR with a high affinity (Ki=47 nM) and selectivity and has a high potency towards CYP17A1 (IC50=22 nM). In addition, ODM-204 inhibited testosterone-mediated nuclear translocation of AR and the mutant ARs (IC50 values for AR(T877A), AR(W741L), and AR(F876L) were 95, 277, and 6 nM, respectively), and suppressed androgen-induced cell proliferation of LNCaP (IC50=170 nM) and VCaP (IC50=280 nM) cells. In a VCaP xenograft model, ODM-204 showed significant antitumor activity (tumor growth inhibition=66%). In rats, inhibitory effects of leuprolide acetate on testosterone production and androgen-sensitive organ weights were potentiated by ODM-204. Conclusions: ODM-204 is a promising new dual CYP17A1 and AR inhibitor for the treatment of CRPC. Clinical trials in patients with mCRPC will be started in early 2015.

The use of baculovirus vectors shows promise as a tool for gene delivery into mammalian cells. These insect viruses have been shown to transduce a variety of mammalian cell lines, and gene transfer has also been demonstrated in vivo. In this study, we generated two recombinant baculovirus vectors displaying an integrin-specific motif, RKK, as a part of two different loops of the green fluorescent protein (GFP) fused with the major envelope protein gp64 of Autographa californica M nucleopolyhedrovirus. By enzyme linked immunosorbent assays, these viruses were shown to bind a peptide representing the receptor binding site of an α2 integrin, the α2I-domain. However, the interaction was not strong enough to overcome binding of wild type gp64 to the unknown cellular receptor(s) on the surface of α2 integrin-expressing cells (CHO-α2β1) or enhance the viral uptake. After treatment of these cells with phospholipase C, internalization of all viruses was blocked or decreased significantly. However, one of the RKK displaying viruses, AcGFP(K)gp64, was still able to internalize into CHO-α2β1 cells, although at a lower level as compared to non-treated cells. This may indicate the possible utilization of a PLC independent alternative route via, in this case, the α2β1 integrin.

Baculovirus vectors have been shown to enter a variety of mammalian cell lines and gene transfer with wild-type baculovirus (WT) has been demonstrated both in vitro and in vivo. Different protein motifs have been displayed on the viral surface to serve as ligands for cell-specific receptor molecules. We have generated recombinant baculovirus vectors displaying an RGD-motif, recognized by alphaV integrin, on the viral surface. The RGD motifs within the C-terminus of coxsackie virus A9 and human parechovirus 1 VP1 proteins were fused to the N-terminus of the major envelope glycoprotein, gp64, of Autographa californica multiple nucleopolyhedrovirus. The recombinant RGD-presenting viruses bound more efficiently to the surface of human lung carcinoma cells (A549), known to contain alphaV integrins, as compared to WT baculovirus. In addition, the binding pattern of the RGD-displaying baculovirus showed extensive clustering. This most likely represents clustering of the integrin molecules on the cell surface, induced by binding of the RGD-displaying baculovirus. Finally, the transduction efficiency of an RGD-representing virus increased by almost three-fold as monitored by light emission measurements. In conclusion, these results suggest that the RGD-motif is functional on the surface of baculovirus and thereby these tropism-modified viruses bind more efficiently as well as enhance the transduction efficiency of human cancer cells expressing alphaV integrins.

The collagens are recognized by the alphaI domains of the collagen receptor integrins. A common structural feature in the collagen-binding alphaI domains is the presence of an extra helix, named helix alphaC. However, its participation in collagen binding has not been shown. Here, we have deleted the helix alphaC in the alpha(2)I domain and tested the function of the resultant recombinant protein (DeltaalphaCalpha(2)I) by using a real-time biosensor. The DeltaalphaCalpha(2)I domain had reduced affinity for type I collagen (430 +/- 90 nM) when compared with wild-type alpha(2)I domain (90 +/- 30 nM), indicating both the importance of helix alphaC in type I collagen binding and that the collagen binding surface in alpha(2)I domain is located near the metal ion-dependent adhesion site. Previous studies have suggested that the charged amino acid residues, surrounding the metal ion-dependent adhesion site but not interacting with Mg(2+), may play an important role in the recognition of type I collagen. Direct evidence indicating the participation of these residues in collagen recognition has been missing. To test this idea, we produced a set of recombinant alpha(2)I domains with mutations, namely D219A, D219N, D219R, E256Q, D259N, D292N, and E299Q. Mutations in amino acids Asp(219), Asp(259), Asp(292), and Glu(299) resulted in weakened affinity for type I collagen. When alpha(2) D219N and D292N mutations were introduced separately into alpha(2)beta(1) integrin expressed on Chinese hamster ovary cells, no alterations in the cell spreading on type I collagen were detected. However, Chinese hamster ovary cells expressing double mutated alpha(2) D219N/D292N integrin showed remarkably slower spreading on type I collagen, while spreading on type IV collagen was not affected. The data indicate that alpha(2)I domain binds to type I collagen with a different mechanism than to type IV collagen.

Although the benefits of contact for positive intergroup relations are widely acknowledged, less is known about how group members construct the agency and responsibility of contact participants in intergroup encounters. Using critical discursive psychology, we analysed the interpretative repertoires that Finnish majority mothers (N = 13) and mothers with an immigrant background (N = 10) used when talking about a hypothetical intergroup encounter among Finnish and immigrant mothers in a ‘family café’ (a group for mothers and children). Our analysis identified five interpretative repertoires that differed in terms of the levels of categorization used (individual, group, motherhood) and how agency and responsibility for initiating contact were discursively attributed to the parties in the intergroup encounter. Overall, constructing someone as agentic did not automatically result in their being portrayed as more responsible for making contact. Respondents described contact to occur with only two repertoires, in which both agency and responsibility for initiating contact were discursively attributed to the same party. This highlights the need to consider both agency and sense of responsibility as possible factors preceding intergroup contact. publishedVersion

Informal segregation has been widely studied in various public settings but not on public playgrounds. Drawing on an 11-month ethnography among mothers of young children, we examine how informal segregation is (re)produced on public playgrounds in two ethnically diverse neighborhoods in Finland. Our findings reveal different normative practices. First, normative rhythms and parenting practices structure playground activities by limiting opportunities for contact between ethnic minority and majority groups and producing exclusive spaces. Second, group norms and the seeking of ethnic/racial ingroup members together regulate mothers’ interaction with outgroup mothers on playgrounds; mothers are inclined toward their ingroup while outgroup mothers are often ignored, resulting in only illusory contact. Based on our analysis, we argue that by better understanding the normative roots of segregation, more comprehensive and effective interventions can be designed to facilitate positive contact in this population. publishedVersion