Objective:Radioimmunotherapy (RIT) is a combination of the therapeutic properties of radioisotopes and specific antibodies used to eliminate tumors. 90Y-OTSA101 was developed by labeling Frizzled homologue 10 (FZD10) antibody with Yttrium-90 (90Y). Because cervical cancer exhibits good radiosensitivity, the present study aimed to assess for prevalence of FZD10 in cervical cancer tissue and investigate the potential of 90Y-OTSA101 treatment for cervical cancer. Methods:The expression of FZD10 was evaluated via immunohistochemistry using formalin-fixed paraffin blocks collected from 84 patients with cervical cancer who underwent surgery. Normal cervical from 10 patients, normal ovarian tissues from 10, uterine carcinomas from 10, and uterine sarcomas from 9 were used for comparison.Mice inoculated with the SiHa cell line, which is derived from cervical squamous cell carcinoma, were used to examine the effects of 90Y-OTSA101. Results:The evaluation results indicated that 70 (91 %) of the cervical cancer cases expressed FZD10 more frequently than 3 (30 %), 1 (10 %), 2 (20 %), and 0 (0 %) of the normal cervix, normal ovary, uterine carcinoma, and uterine sarcoma cases, respectively. The group treated with 3.7 MBq of 90Y-OTSA101 demonstrated greater antitumor effects, defined as tumor shrinkage or growth suppression, than the group treated with 1.85 MBq. No significant difference in body weight was observed between the groups during the observation period. Conclusions:FZD10 may be an appropriate target and 90Y-OTSA101 may have potential for the treatment of cervical cancer.

To select the most suitable chelate for 225 Ac radiolabeling of the anti-FZD10 antibody OTSA101, we directly compared three chelates: S-2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane tetraacetic acid (p-SCN-Bn-DOTA), 2,2',2″-(10-(1-carboxy-4-((4-isothiocyanatobenzyl)amino)-4-oxobutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl) triacetic acid (p-SCN-Bn-DOTAGA), and 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid mono-N-hydroxysuccinimide ester (DO3A-NHS-ester). We evaluated the binding affinity of the chelate-conjugated OTSA101 antibodies, as well as the labeling efficiency and stability in murine serum of 225 Ac-labeled OTSA101 as in vitro properties. The biodistribution, intratumoral distribution, absorbed doses, and therapeutic effects of the chelate-conjugated OTSA101 antibodies were assessed in the synovial sarcoma mouse model SYO-1. Of the three conjugates, DOTAGA conjugation had the smallest impact on the binding affinity (p < 0.01). The labeling efficiencies of DOTAGA-OTSA101 and DO3A-OTSA101 were 1.8-fold higher than that of DOTA-OTSA101 (p < 0.01). The stabilities were similar between 225 Ac-labeled DOTA-OTSA101, DOTAGA-OTSA101, and DO3A-OTSA101in serum at 37 and 4°C. The dosimetric analysis based on the biodistribution revealed significantly higher tumor-absorbed doses by 225 Ac-labeled DOTA-OTSA101 and DOTAGA-OTSA101 compared with 225 Ac-DO3A-OTSA101 (p < 0.05). 225 Ac-DOTAGA-OTSA101 exhibited the highest tumor-to-bone marrow ratio, with bone marrow being the dose-limiting tissue. The therapeutic and adverse effects were not significantly different between the three conjugates. Our findings indicate that among the three evaluated chelates, DOTAGA appears to be the most promising chelate to produce 225 Ac-labeled OTSA101 with high binding affinity and high radiochemical yields while providing high absorbed doses to tumors and limited absorbed doses to bone marrow.

Supplementary Table 1 from Cell-Permeable Peptide DEPDC1-ZNF224 Interferes with Transcriptional Repression and Oncogenicity in Bladder Cancer Cells

Supplementary Figure Legends 1-10 from Cell-Permeable Peptide DEPDC1-ZNF224 Interferes with Transcriptional Repression and Oncogenicity in Bladder Cancer Cells

Supplementary Figures 2-6 from Cell-Permeable Peptide DEPDC1-ZNF224 Interferes with Transcriptional Repression and Oncogenicity in Bladder Cancer Cells

Supplementary Methods from Cell-Permeable Peptide DEPDC1-ZNF224 Interferes with Transcriptional Repression and Oncogenicity in Bladder Cancer Cells

Synovial sarcomas are rare tumors arising in adolescents and young adults. The prognosis for advanced disease is poor, with an overall survival of 12-18 months. Frizzled homolog 10 (FZD10) is overexpressed in most synovial sarcomas, making it a promising therapeutic target. The results of a phase 1 trial of β-radioimmunotherapy (RIT) with the 90 Y-labeled anti-FZD10 antibody OTSA101 revealed a need for improved efficacy. The present study evaluated the potential of α-RIT with OTSA101 labeled with the α-emitter 225 Ac. Competitive inhibition and cell binding assays showed that specific binding of 225 Ac-labeled OTSA101 to SYO-1 synovial sarcoma cells was comparable to that of the imaging agent 111 In-labeled OTSA101. Biodistribution studies showed high uptake in SYO-1 tumors and low uptake in normal organs, except for blood. Dosimetric studies showed that the biologically effective dose (BED) of 225 Ac-labeled OTSA101 for tumors was 7.8 Bd higher than that of 90 Y-labeled OTSA101. 90 Y- and 225 Ac-labeled OTSA101 decreased tumor volume and prolonged survival. 225 Ac-labeled OTSA101 achieved a complete response in 60% of mice, and no recurrence was observed. 225 Ac-labeled OTSA101 induced a larger amount of necrosis and apoptosis than 90 Y-labeled OTSA101, although the cell proliferation decrease was comparable. The BED for normal organs and tissues was tolerable; no treatment-related mortality or obvious toxicity, except for temporary body weight loss, was observed. 225 Ac-labeled OTSA101 provided a high BED for tumors and achieved a 60% complete response in the synovial sarcoma mouse model SYO-1. RIT with 225 Ac-labeled OTSA101 is a promising therapeutic option for synovial sarcoma.

Synovial sarcoma ( SS ) is a rare yet refractory soft‐tissue sarcoma that predominantly affects young adults. We show in a mouse model that radioimmunotherapy ( RIT ) with an α‐particle emitting anti‐Frizzled homolog 10 ( FZD 10) antibody, synthesized using the α‐emitter radionuclide astatine‐211 ( 211 At‐ OTSA 101), suppresses the growth of SS xenografts more efficiently than the corresponding β‐particle emitting anti‐ FZD 10 antibody conjugated with the β‐emitter yettrium‐90 ( 90 Y‐ OTSA 101). In biodistribution analysis, 211 At was increased in the SS xenografts but decreased in other tissues up to 1 day after injection as time proceeded, albeit with a relatively higher uptake in the stomach. Single 211 At‐ OTSA 101 doses of 25 and 50 μCi significantly suppressed SS tumor growth in vivo, whereas a 50‐μCi dose of 90 Y‐ OTSA 101 was needed to achieve this. Importantly, 50 μCi of 211 At‐ OTSA 101 suppressed tumor growth immediately after injection, whereas this effect required several days in the case of 90 Y‐ OTSA 101. Both radiolabeled antibodies at the 50‐μCi dosage level significantly prolonged survival. Histopathologically, severe cellular damage accompanied by massive cell death was evident in the SS xenografts at even 1 day after the 211 At‐ OTSA 101 injection, but these effects were relatively milder with 90 Y‐ OTSA 101 at the same timepoint, even though the absorbed doses were comparable (3.3 and 3.0 Gy, respectively). We conclude that α‐particle RIT with 211 At‐ OTSA 101 is a potential new therapeutic option for SS .

Cancer is the second most common cause of death in US in 2015 and has been the leading cause of death in Japan since 1981. Despite of improved therapeutic strategies in cancer, only limited treatment options can still be available to a subset of patients. Many efforts have been made to develop molecular targeted drugs, but the success rate did not reach to patients’ expectation. Unlike the conventional chemotherapy, the use of maximum tolerated dose (MTD) and the measurement of bulk tumor volume are not always appropriate for the clinical evaluation of molecular targeted drugs. To optimize and personalize the dosing for targeted agents, detailed molecular pathway and biomarkers that are affected by the drug should be elucidated. OTS167 was developed as a novel and potent MELK kinase inhibitor that we previously reported. The expression of MELK is elevated in various human cancers both in solid and hematological tumor and MELK is known to be associated with cancer progression and poor prognosis. Because MELK is indicated its critical roles in cancer stem cell proliferation as well, targeting MELK is an attractive and promising therapeutic strategy for cancer patient. Here, we report the molecular mechanism of action of OTS167 in preclinical model. OTS167 suppressed MELK activity and promoted MELK protein degradation by inhibition of autophosphorylation. OTS167-treated cells showed drastic morphological transformation with the induction of p53 and p21 expression. We also investigated the expression of stem cell markers to elucidate whether OTS167 suppresses cancer stem-like properties through inhibition of MELK. Furthermore, we evaluated antitumor activity of OTS167 using human xenograft model and molecular changes in tumor tissue. The expression of MELK and downstream molecules were decreased in OTS167-treated xenograft tumor tissues by IHC. The change of expression and pharmacological effect were well correlated. Our data provide the evidence for the concept that OTS167 suppresses tumor growth through the inhibition of MELK signaling pathway and suggest the possibility of biomarkers for the assessment of clinical efficacy. Citation Format: Suyoun Chung, Kyoko Kijima, Yosuke Harada, Naofumi Takamatsu, Takashi Miyamoto, Yo Matsuo, Yusuke Nakamura. Pharmacologic, pharmacodynamic action of MELK kinase inhibitor OTS167 in cancer cells. [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 2158.

MELK is upregulated in various types of human cancer and is known to be associated with cancer progression, maintenance of stemness, and poor prognosis. OTS167, a MELK kinase inhibitor, shows potent growth-suppressive effect on human tumors in a xenograft model, but the detailed mode of action has not been fully elucidated. In this study, we demonstrate the molecular mechanism of action of MELK inhibitor OTS167 in a preclinical model. OTS167-treated cells caused morphological transformation, induced the differentiation markers, and reduced stem-cell marker expression. Furthermore, we identified DEPDC1, known as an oncogene, as an additional downstream molecule of the MELK signaling pathway. MELK enhanced DEPDC1 phosphorylation and its stability. The expression of MELK and downstream molecules was decreased in OTS167-treated xenograft tumor tissues, which revealed central necrosis and significant growth suppression. Our data should further shed light on the mechanism of action how OTS167 suppresses tumor growth through the inhibition of the MELK signaling pathway and suggest the possibility of biomarkers for the assessment of clinical efficacy.

Poly(2-pyridone-3,5-diyl)s with -(CH2)(4)-SO3M (M = H or Na) side chains have been prepared by nickel-complex promoted dehalogenative polycondensation. A composite film of the polymer with -(CH2)(4)-SO3H side chains and poly(vinyl alcohol) showed a proton conductivity of 1.5 x 10(-1) S cm(-1) at 80 degrees C and 95% humidity. A copolymer with pyridine showed a high stability against oxidation by a Fenton reagent. (C) 2012 Elsevier Ltd. All rights reserved.

Dibromopyridines or dibromopyridone with –(CH2)m–SO3Na group(s) has been prepared via the reactions of the corresponding dibromopyridines with –OH and –NH2 groups with sultone. These compounds were converted into polymers with the –(CH2)m–SO3H groups via organometallic polycondensation. The polymer showed proton conducting properties and high stability toward oxidation.

AbstractBladder cancer is the second most common genitourinary cancer worldwide, yet its oncogenic origins remain poorly understood. The cancer-testis antigen DEPDC1 was shown recently to contribute to bladder cancer oncogenesis. In this study, we examined the biological functions of DEPDC1 and defined a potential therapeutic strategy to target this molecule. Coimmunoprecipitation and immunocytochemistry revealed that DEPDC1 interacted and colocalized with zinc finger transcription factor ZNF224, a known transcriptional repressor. Inhibiting this interaction with a cell-permeable peptide corresponding to the ZNF224-interacting domain in DEPDC1 induced apoptosis of bladder cancer cells in vitro and in vivo. By inhibiting DEPDC1-ZNF224 complex formation, this peptide triggered transcriptional activation of A20, a potent inhibitor of the NF-κB signaling pathway. Our findings indicate that the DEPDC1-ZNF224 complex is likely to play a critical role in bladder carcinogenesis. Cancer Res; 70(14); 5829–39. ©2010 AACR.

In fuel cells applications, current-ripple reduction is essential for conversion efficiency and life span. This paper analyzes the pulse-link DC-AC converter for fuel cells applications operated in zero-current-slope mode. As the result, in zero-current-slope operation mode, input-current-ripple is reduced. Furthermore, in this operation mode, the parameters of series LC circuit which is worked as ripple canceling are less values.

Background and objective:Whether beta(2)-adrenoceptor gene (ADRB2) polymorphisms are associated with airway responsiveness to beta(2)-agonist medications remains controversial, partly due to factors that may confound pharmacogenetic associations, including age, cigarette smoking and airway remodelling. To overcome these problems, we performed an analysis using parameters that reflected the specific bronchodilator response to beta(2)-agonists.Methods:The increases in FEV1 after inhalation of procaterol hydrochloride (Delta FEV1 procaterol) or oxitropium bromide (Delta FEV1 oxitropium), and after sequential inhalation of procaterol and oxitropium (total airway reversibility), were measured in 81 Japanese patients with moderate to severe asthma. Approximately 3 kb of the DNA sequence of the coding and 5'-flanking regions of ADRB2 were genotyped by direct sequencing and PCR-restriction fragment length polymorphism assay.Results:The mean age of the participants was 54 years, and 38 (47%) were smokers. Although Delta FEV1 procaterol and Delta FEV1 oxitropium adjusted for predicted FEV1 were not associated with ADRB2 polymorphisms, the ratio of Delta FEV1 procaterol to total airway reversibility was significantly associated with the ADRB2 A46G genotype (P < 0.05). Patients who were homozygous for the A46 allele (arginine at amino acid 16) were more responsive than carriers of the G46 (glycine 16) allele (P = 0.008). Multivariate linear regression analysis showed that Delta FEV1 procaterol was correlated with the number of A46 alleles (P = 0.014), and also with total airway reversibility (P < 0.001) and smoking index in current smokers (P = 0.009).Conclusions:The ADRB2 A46G polymorphism was associated with a relatively greater bronchodilator responsiveness to beta(2)-agonists even in elderly asthmatic patients and smokers.

Excited state characteristics of aza analogues of nucleic acid bases, 8-azaadenine (8AA), 5-azacytosine (5AC), 8-azaguanine (8AG), and 6-azauracil (6AU), in acetonitrile solution were comprehensively investigated with steady state absorption and emission spectra, transient absorption measurements, emission measurements for the singlet oxygen molecule, and time-dependent density functional theory (TD-DFT) calculations. The triplet-triplet absorption spectrum of 8AA whose peak was 455 nm was observed for the first time. Sensitized singlet oxygen formation of 8AA was also observed in O(2)-saturated acetonitrile with quantum yields of 0.15 +/- 0.02. It was concluded that there were two kinds of aza analogues of nucleic acid bases: type A had substantial quantum yield for the intersystem crossing and potential of O2 (1Delta(g)) formation (8AA and 6AU), and type B did not (5AC and 8AG). TD-DFT calculations indicated that type A molecules had a dark 1npi* state below the first allowed 1pipi* state, while both S1 and S2 states for type B molecules had a pipi* character. It strongly suggested that the dark 1npi* state below the 1pipi* state would play an important role in the ISC process of aza analogues of nucleic acid bases.

Reactions of 2-amino-3,5-dibromopyridine with sultones gave 3,5-dibromopyridines with –NH(CH2)mSO3Na (m = 3 and 4) groups at the 2-position of pyridine. The obtained compound served as a starting m...

This paper examines the effect of input current-ripple reduction on Pulse-link DC-AC converter for fuel cells applications. For reducing the input current-ripple, series LC circuit has been connected in parallel at the Pulse-link DC-AC converter. The input current-ripple characteristics have two main domains with combination of series LC parameters. The current-ripple level depends on inductance values. Input current-ripple is reduced when the inductor current flown at series LC circuit has zero-slopes in one switching period. At that time, small parameters values can be reached.

This paper mentions the input current ripple reduction method of the Pulse-link DC-AC Converter for Fuel Cells. The conventional DC-AC converter for fuel cells is interpolated large capacitor between boost converter stage and PWM inverter stage. That capacitor disturbs the size reduction of this unit. To overcome this problem, authors have proposed a novel topology called as Pulse-link DC-AC converter. The proposed topology provides boosted-voltage pulse directly to PWM inverter. This topology does not require large capacitor between two stages. Instead, small values of inductor and capacitor are connected series and inserted between two stages in parallel. This paper examines the relationship between the inductor and capacitor values and input current-ripple. As the result, inductor value has the relationship with current-ripple.