The diacylglycerol kinase ζ inhibitor ASP1570 augments natural killer cell function.

The enhancement of T cell and NK cell function is an immunotherapeutic strategy for combating cancer. Antibodies that block inhibitory receptors, such as PD-1 and CTLA4, augment T cell function and have been successful in curing patients with some types of cancer. As an alternative approach to targeting specific inhibitory receptors by antibodies, small molecule drugs that inhibit negative regulators of T cell activation have been sought. One potential pharmacological target is diacylglycerol (DAG) kinase (DGK)ζ, which is an enzyme that acts as a negative regulator of DAG by phosphorylating DAG and converting it into phosphatidic acid. DAG-mediated signaling is critical for T cell activation through its T cell receptor and NK cell activation downstream of a variety of activating receptors. Thus, DGKζ-deficient T cells and NK cells display increased function upon activating receptor engagement. Moreover, treatment with the DGKζ-selective inhibitor ASP1570 augments T cell function. In this study, we sought to test whether the acute inhibition of DGKζ by ASP1570 augments NK cell function. We find that ASP1570 enhances DAG-mediated signaling in immunoreceptor-stimulated NK cells. Accordingly, ASP1570 treatment enhanced IFNγ production and degranulation of immunoreceptor-activated NK cells in vitro and NK cell-mediated tumor clearance in vivo. Thus, ASP1570 enhances both T and NK cell function, which could possibly induce more durable anti-tumor responses for immunotherapy.

Manipulation of diacylglycerol and ERK-mediated signaling differentially controls CD8+ T cell responses during chronic viral infection.

Introduction: Activation of T cell receptor (TCR) signaling is critical for clonal expansion of CD8+ T cells. However, the effects of augmenting TCR signaling during chronic antigen exposure is less understood. Here, we investigated the role of diacylglycerol (DAG)-mediated signaling downstream of the TCR during chronic lymphocytic choriomeningitis virus clone 13 (LCMV CL13) infection by blocking DAG kinase zeta (DGKζ), a negative regulator of DAG. Methods: We examined the activation, survival, expansion, and phenotype of virus-specific T cell in the acute and chronic phases of LCMV CL13-infected in mice after DGKζ blockade or selective activation of ERK. Results: Upon LCMV CL13 infection, DGKζ deficiency promoted early short-lived effector cell (SLEC) differentiation of LCMV-specific CD8+ T cells, but this was followed by abrupt cell death. Short-term inhibition of DGKζ with ASP1570, a DGKζ-selective pharmacological inhibitor, augmented CD8+ T cell activation without causing cell death, which reduced virus titers both in the acute and chronic phases of LCMV CL13 infection. Unexpectedly, the selective enhancement of ERK, one key signaling pathway downstream of DAG, lowered viral titers and promoted expansion, survival, and a memory phenotype of LCMV-specific CD8+ T cells in the acute phase with fewer exhausted T cells in the chronic phase. The difference seen between DGKζ deficiency and selective ERK enhancement could be potentially explained by the activation of the AKT/mTOR pathway by DGKζ deficiency, since the mTOR inhibitor rapamycin rescued the abrupt cell death seen in virus-specific DGKζ KO CD8+ T cells. Discussion: Thus, while ERK is downstream of DAG signaling, the two pathways lead to distinct outcomes in the context of chronic CD8+ T cell activation, whereby DAG promotes SLEC differentiation and ERK promotes a memory phenotype.

Randomized, Double-Blind, Controlled Study to Evaluate Safety and Pharmacokinetics of Single Ascending Doses of ASP5354, an Investigational Imaging Product, in Healthy Adult Volunteers.

Intraoperative ureter identification helps reduce the risk of ureteral injury. Currently, no suitable agents for real-time ureter visualization are approved. ASP5354 (TK-1) is a novel indocyanine green derivative. In this first-in-human phase 1, double-blind, sequential ascending-dose study, urethral catheters were placed in 6 healthy volunteers who were randomized to single-dose, intravenous ASP5354 0.1 mg (n = 4) or placebo (n = 2). Sequential dose escalations to 0.5-, 2-, 8-, and 24-mg ASP5354 in new cohorts were contingent upon Dose-Escalation Committee approval after review of pharmacokinetic (PK) and safety data. Blood and urine samples were collected over 24 hours following dose administration. Objectives were to assess the safety/tolerability and PK of ASP5354. Treatment-emergent adverse events (TEAEs) were reported in 3 (15%) and 2 (20%) participants in the ASP5354 and placebo groups, respectively. In the former, there were 6 TEAEs (5/6 grade 1-2). One ASP5354 participant experienced grade 3 pyelonephritis, attributed to the catheter. No TEAEs were related to ASP5354. Mean plasma terminal elimination half-life ranged from 2.1 to 3.6 hours, with near complete urinary excretion of unchanged ASP5354 within 24 hours after administration. Linear and dose-proportional PK were observed. These results support further evaluation of ASP5354 at doses up to 24 mg for intraoperative near-infrared fluorescence ureter visualization.

Investigation of Safety and Tolerability of ASP3652 Based on Clinical Studies of Cerebrospinal Fluid Transfer After Multiple Doses and Exposure After Single Doses at High Dose Levels.

INTRODUCTION: The studies described here were conducted to investigate the central nervous system (CNS) transfer of ASP3652, a peripherally acting inhibitor of fatty acid amide hydrolase, after multiple doses at around the anticipated therapeutic dose and the safety, tolerability, and pharmacokinetics after single doses at corresponding supratherapeutic doses in healthy subjects. METHODS: Study 1 was an open-label multiple dose study in which ASP3652 (300 mg bid) or matching placebo was administered in multiple doses to healthy subjects. Study 2 was a placebo-controlled, randomized 4 × 4 crossover study in which ASP3652 was given as three single ascending doses of ASP3652 (600-1800 mg) or matching placebo to healthy subjects. Levels of ASP3652 and endocannabinoids (eCBs) in plasma, cerebrospinal fluid (CSF) (study 1 only), and safety were evaluated. RESULTS: In study 1, ASP3652 was readily absorbed to reach Cmax at 1 h after dosing. AUCtau and Cmax of ASP3652 in CSF were approximately 0.2% and 0.06% of the AUCtau and Cmax in plasma after multiple doses of ASP3652 300 mg bid. At steady state the area under the response-time curve (AURC) from 0 to 12 h and the maximum response for anandamide in plasma were approximately 550-fold and 230-fold higher than those in CSF. In study 2, the Cmax and AUC of ASP3652 increased higher than dose proportionally in subjects receiving 600-1800 mg ASP3652. For eCBs, although the AURC increased less than dose proportionally, maximum plasma levels were comparable across all treatment groups. The incidence of adverse events (AEs) was similar across all treatment groups including the placebo group. There was no evidence of CNS-related side effects. CONCLUSIONS: ASP3652 showed low CNS penetration at the anticipated therapeutic dose and was well tolerable without any CNS-related AEs at supratherapeutic doses, supporting that the drug can be safely tested at the anticipated therapeutic dose. TRIAL REGISTRATION: ClinicalTrials.gov identifier, NCT02034734 for study 1, NCT01815684 for study 2.

The Safety, Tolerability, Pharmacokinetics, and Pharmacodynamics of ASP3652 in First-in-Human and Ascending Multiple Oral Dose Studies in Healthy Subjects.

INTRODUCTION: Inhibitors of fatty acid amide hydrolase (FAAH) increase the levels of endocannabinoids and have shown analgesic and anti-inflammatory activity in animal models. ASP3652 is a peripherally acting FAAH inhibitor in development for the treatment of chronic bladder and pelvic pain disorders. Here we describe the safety, pharmacokinetics, and pharmacodynamics of single and multiple oral doses of ASP3652 administered in healthy non-elderly and elderly male and female volunteers. METHODS: Study 1 was a combined single-ascending dose and food-effect study in which ASP3652 was given as single doses (1-600 mg) or matching placebo in healthy subjects. Study 2 was a multiple ascending dose study in which ASP3652 or matching placebo was administered in multiple oral doses (10-300 mg bid and 600 mg qd for 14 days) to healthy subjects. In both studies, the levels of ASP3652, FAAH, endocannabinoids (eCBs) and safety were evaluated. RESULTS: ASP3652 was readily absorbed to reach Cmax at 1 h after a single dose. Steady state was reached within 3 days after the start of multiple dosing. The Cmax and AUC of ASP3652 increased in a slightly more than dose-proportional manner after a single dose of ASP3652 at 30-600 mg. There was some accumulation (15-38%) based on Cmax and AUC12h upon multiple doses. Cmax was 47% lower in combination with food. There was no significant effect of gender or age on the pharmacokinetics of ASP3652. FAAH activity was inhibited in a dose-dependent manner in all dose groups after single and multiple doses of ASP3652, paralleled by an increase in plasma levels of anandamide (AEA). The incidence of adverse events following multiple doses was similar across all treatment groups including the placebo group. CONCLUSIONS: Single and multiple doses of ASP3652 were safe and well tolerated and increased endogenous cannabinoid plasma levels.

Safety, Tolerability, Pharmacokinetics, and Pharmacodynamics of ASP3652, a Reversible Fatty Acid Amide Hydrolase Inhibitor, in Healthy, Nonelderly, Japanese Men and Elderly, Japanese Men and Women: A Randomized, Double-blind, Placebo-controlled, Single and Multiple Oral Dose, Phase I Study.

PURPOSE: This study aimed to evaluate the safety, tolerability, and pharmacokinetic and pharmacodynamic properties of ASP3652, a peripherally acting inhibitor of peripheral fatty acid amide hydrolase (FAAH) after 30-, 100-, 300-, 600-, and 900-mg single and 100- and 300-mg BID multiple oral dose in Japanese patients. METHODS: This was a randomized, double-blind, placebo-controlled, single and multiple oral dose Phase I study in healthy, nonelderly men and elderly men and women. The study consisted of 2 parts: in the single oral dose part, 40 healthy, nonelderly men were randomized to receive placebo or ASP3652; in the multiple oral dose part, 48 enrolled nonelderly men and elderly men and women were randomized to receive placebo or ASP3652. In both parts, the investigator judged whether the individuals were healthy based on the results of physical examinations and screening. The safety profile was assessed by examining adverse events, defined as any untoward medical occurrence in an individual administered the study drug and that did not necessarily have a causal relationship with the study treatment; clinical laboratory evaluations; vital signs; the Profile of Mood States scale; and standard 12-lead ECGs and 12-lead ECGs for QT assessment. Pharmacokinetic parameters were estimated using unchanged ASP3652 concentrations in plasma and urine. Pharmacodynamic parameters were estimated using FAAH activity and plasma anandamide, oleoylethanolamide, and palmitoylethanolamide concentrations. Safety and tolerability profiles were compared with the placebo group. FINDINGS: ASP3652 was rapidly absorbed to reach Cmax in a single dose and near steady-state at approximately 3 days after the start of multiple dosing. The Cmax and AUC of ASP3652 were slightly higher than dose proportional after a single dose of ASP3652 at 30-900 mg. There was no apparent accumulation based on Cmax and AUC0-12 after multiple doses. Although no differences were found in Cmax or AUC0-12 by age in men, Cmax and AUC0-12 were slightly higher in elderly women than elderly men. FAAH activity was inhibited in a dose-dependent manner, and plasma levels of anandamide, oleoylethanolamide, and palmitoylethanolamide increased in all dose groups after single and multiple doses of ASP3652. The incidence of adverse events after multiple doses, which ranged from 44.4% to 66.7%, was similar across all treatment groups, including the placebo group. IMPLICATIONS: Single and multiple doses of ASP3652 were well tolerated and increased endogenous cannabinoids.

Telecommunication system for children undergoing stem cell transplantation.

BACKGROUND: Isolation in a germ-free unit is a stressful experience for pediatric patients undergoing hematopoietic stem cell transplantation (HSCT). To reduce the psychological distress of such children, a Web-based telecommunications system was developed. METHODS: The authors developed a telecommunication system that linked a laminar air flow (LAF) room that had a high efficiency particulate air filter with the hospital school/patients' homes via the Internet. Fifteen children isolated in the LAF room for allogeneic HSCT were enrolled in this study. The present study evaluated whether the system was feasible for the patients during the acute phase of HSCT. RESULTS: In 10 patients, the proportion of days when they telecommunicated with teachers and/or other patients in the hospital school was 64.6 ± 32.3%. The telecommunication with the hospital school facilitated the continuation of school study under teachers' guidance, reducing the problem of lost schooling. In 13 patients, the proportion of days when they telecommunicated with their homes was 68.0 ± 34.8%. Ten of them frequently telecommunicated with their family members (especially siblings), and three patients called out to their pets at home. The incidence of telecommunication on the days when the patients had HSCT-related symptoms including vomiting did not differ from that of telecommunication on the days when no symptoms were evident. CONCLUSIONS: A telecommunication system linked to a hospital school and/or the patients' homes is feasible for children undergoing HSCT, and may improve their health-related quality of life. A larger, prospective study is required to evaluate whether the telecommunication system can reduce HSCT-associated psychological and psychiatric symptoms.

[Evaluation of the accuracy of a compensating filter based on compact NC-Mill for radiation therapy].

The present report describes the fabrication technique and dosimetry aspects of a desktop numerically controlled milling machine (NC-Mill) based a compensator system that uses lead clay (Shield cray, Reactor Experiments, Inc., U.S.A.). Effective path lengths of patients were determined for CT image sets using the ray-tracing technique and converted to compensator thickness with the equivalent TMR method. Rigid urethane foam was processed with the NC-Mill to produce a mold for filters, and the lead clay was adopted as the compensating material. The dose distribution was measured on the compensating plane of an anthropomorphic phantom and a stair-step PMMA phantom. It was found that the radiation field with inhomogeneous dose was as high as 30%+/-3% with the compensating filters. In addition, when the absorbed dose at the central axis of 52 compensating filters that were used clinically was measured, 75.0% showed an error of less than +/-3%, and 3.8% showed the maximum dose error: >+/-5%. Overall, the present system was capable of producing dose uniformity to within +/-5% for a stair-step phantom, an anthropomorphic phantom, and clinical situations.

Realtime ultrasound screening by satellite telecommunication.

Realtime ultrasound screening was carried out using a medical examination vehicle equipped with a diagnostic ultrasound scanner and a satellite telecommunication system. Screening was performed on 205 residents in a rural community in Japan and consisted of 57 cardiac, 57 abdominal, 60 thyroid and 31 breast ultrasound scans. The resolution of the realtime ultrasound images sent via communications satellite at 1.5 Mbit/s was almost identical to that of the original images taken by the ultrasound scanner. A disorder was diagnosed in 11 (19%) of the cardiac examinations, 28 (49%) of the abdominal, 19 (32%) of the thyroid and 7 (23%) of the breast examinations. Although some technical problems occurred, ultrasound screening by telemedicine appears to be a promising technique for those who live in rural communities.

[Network system for alpine ambulance using long distance wireless LAN and CATV LAN].

A network of the emergency telemedicine for mountain climbers in Japan Alps area has been developed. The network consists of 2.4GHz wireless LAN with over 30 km distance to be connected the Cable TV LAN. We confirmed that this network is able to transfer the high quality video and audio signal suitable for the telemedicine between the mountain huts and hospitals. In this paper, we pointed out the shortcomings of wireless LAN system and cable TV network system in use for the telemedicine, and wireless LAN module covered more than 30km and an extension system of upstream frequency bandwidth for Cable TV network. Experiments suggested that a low cost and long distance wireless LAN is useful for mountain climber's emergency telemedicine.

[Inter-hospital teledermatology conference using a videophone network].

We organized an inter-hospital teledermatology conference with images through a digital telephone line (ISDN, 128kbps). Our conference system consists of two videophones. A videophone "Phoenix PICSEND-R" (NTT) in the multi-point connection mode is used for discussion among doctors from five hospitals. In addition, using another videophone "Phoenix mini type-S" (NTT), clinical and dermoscopic findings obtained with a digital video camera and a videomicroscope "VMS-1900" (Scalar, Tokyo) respectively were transmitted to Shinshu University Hospital from four rural sites. We have found that the quality of the images obtained is entirely sufficient to make an accurate diagnosis. To calibrate the color and size of skin lesions, a matching sticker "CASMATCH" (Bearmedic, Tokyo) has proved useful. Forty-one times of conference were held from September 1999 to August 2002. In total, the concordance rate between the telediagnosis and the finally confirmed diagnosis was 69% (77 of 112 cases). In cases limited to pigmented skin lesions, the rate was higher (84%) and it was concluded that such conditions are suited for the telediagnosis. The ease with which the data can be manipulated enables us to discuss difficult cases in detail. This style of teleconference system may have benefits for dermatologists especially at rural hospitals in consultation with senior doctors.

[Medical support for Belarus after Chernobyl accident using a telemedicine system].

To achieve a close relationship among Gomel Regional Hospital (Gomel, Belarus), Belarusian Children Center of Hematology/Oncology (Minsk, Belarus) and Shinshu University Hospital, we established a telemedicine system using the Inmarsat satellite. The system consists of a TV conference system, a digital microscopic imaging system and a high-definition image server/viewer network system (DICOM). The detailed case conference is possible among three areas of physicians.

[Mobile hospital -real time mobile telehealthcare system with ultrasound and CT van using high-speed satellite communication-].

A real-time telescreening system is developed to detect early diseases for rural area residents using two types of mobile vans with a portable satellite station. The system consists of a satellite communication system with 1.5Mbps of the JCSAT-1B satellite, a spiral CT van, an ultrasound imaging van with two video conference system, a DICOM server and a multicast communication unit. The video image and examination image data are transmitted from the van to hospitals and the university simultaneously. Physician in the hospital observes and interprets exam images from the van and watches the video images of the position of ultrasound transducer on screenee in the van. After the observation images, physician explains a results of the examination by the video conference system. Seventy lung CT screening and 203 ultrasound screening were done from March to June 2002. The trial of this real time screening suggested that rural residents are given better healthcare without visit to the hospital. And it will open the gateway to reduce the medical cost and medical divide between city area and rural area.

Telemedicine of the heart: real-time telescreening of echocardiography using satellite telecommunication.

The early detection of disease is important to keep people healthy, but it is not easy for elderly residents living in rural areas at a distance from well-equipped medical institutes to be examined. Therefore, the present study evaluated the usefulness of real-time telescreening of echocardiography using satellite communications. The participants in this study were 57 residents of a rural community in Japan. The resolution of the real-time ultrasound images sent by communications satellite was almost identical to that of the original images taken in the hospital. Although some problems remain to be solved, telescreening of echocardiography will be a practical method for public health in the near future.

[Telemedicine support system in home care of patients with chronic respiratory failure: preliminary results].

We report the preliminary results of an advanced telemedicine system for the delivery of home care services. We have been trying out a system for a ternary telemedicine network for remote consultation in patients with chronic respiratory failure. The elements of this system are: the patients at home, the home doctor (and/or community hospital), and an institution with respiratory staff (Shinshu University), which were linked by ISDN lines. The system allows real-time visualization, at home, of the patients and their physiological data, which are transmitted simultaneously to both the community hospital and Shinshu University. Respiratory specialists can advise the home doctor on the treatment and follow-up evaluations of distant patients. We consider that the telemedicine network system has the potential to improve clinical outcomes and to provide home care services to patients with chronic health conditions.