Background: It is necessary to make objective and quantitative evaluation of motor disorders caused by brain damage such as stroke.

The chromosome 15q13.3 microdeletion is a pathogenic copy number variation conferring epilepsy, intellectual disability, schizophrenia, and autism spectrum disorder (ASD). We generated mice carrying a deletion of 1.2 Mb homologous to the 15q13.3 microdeletion in human patients. Here, we report that mice with a heterozygous deletion on a C57BL/6 background (D/+ mice) demonstrated phenotypes including enlarged/heavier brains (macrocephaly) with enlarged lateral ventricles, decreased social interactions, increased repetitive grooming behavior, reduced ultrasonic vocalizations, decreased auditory-evoked gamma band EEG, and reduced event-related potentials. D/+ mice had normal body weight, activity levels, sensory gating, and cognitive abilities and no signs of epilepsy/seizures. Our results demonstrate that D/+ mice represent ASD-related phenotypes associated with 15q13.3 microdeletion syndrome. Further investigations using this chromosome-engineered mouse model may uncover the common mechanism(s) underlying ASD and other neurodevelopmental/psychiatric disorders representing the 15q13.3 microdeletion syndrome, including epilepsy, intellectual disability, and schizophrenia.SIGNIFICANCE STATEMENT:Recently discovered pathologic copy number variations (CNVs) from patients with neurodevelopmental/psychiatric disorders show very strong penetrance and thus are excellent candidates for mouse models of disease that can mirror the human genetic conditions with high fidelity. A 15q13.3 microdeletion in humans results in a range of neurodevelopmental/psychiatric disorders, including epilepsy, intellectual disability, schizophrenia, and autism spectrum disorder (ASD). The disorders conferred by a 15q13.3 microdeletion also have overlapping genetic architectures and comorbidity in other patient populations such as those with epilepsy and schizophrenia/psychosis, as well as schizophrenia and ASD. We generated mice carrying a deletion of 1.2 Mb homologous to the 15q13.3 microdeletion in human patients, which allowed us to investigate the potential causes of neurodevelopmental/psychiatric disorders associated with the CNV.

1816 Objectives ASP0777 is a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist as a drug candidate for the treatment of moderate to severe Alzheimer’s disease. The present study was undertaken to confirm brain penetration of [11C]ASP0777 in conscious rhesus macaques. Methods Three male rhesus macaques (6-10 y; 6.850-8.490 kg) were subjected. [11C]ASP0777 was administered either at the dose of tracer alone, 0.1, or 1 mg/kg, and each animal was studied for all doses. A dynamic PET scan of 90 min was performed following an intravenous injection of [11C]ASP0777. Arterial blood samples were collected while PET scanning to measure the ASP0777 concentration in the plasma, which were corrected for metabolites using thin layer chromatography (TLC). Brain distribution was calculated for 12 brain regions as standardized uptake value (SUV), and ASP0777 concentration was calculated according to the specific activity of ASP0777 preparation. Results [11C]ASP0777 showed a good brain penetration and relatively higher uptake in the striatum (SUV = 1.99, brain/plasma = 23.29, at 1 h, tracer dose) and the thalamus (SUV = 2.01, brain/plasma = 23.67, at 1 h, tracer dose). ASP0777 also showed a linear relationship between the dose and brain concentration for all doses tested. Conclusions The penetration of [11C]ASP0777 into the monkey brain could be confirmed which brings useful information for drug discovery research as a kind of pharmacokinetic study.

Receptor tyrosine kinases play a critical role in cell growth, survival, and proliferation, and are considered potential molecular targets for the treatment of cancer. Although several tyrosine kinase inhibitors (TKIs), such as erlotinib and gefitinib, have demonstrated clinical efficacy via the inhibition of the epidermal growth factor receptor (EGFR), most TKIs are only effective in a small proportion of patients. Positron emission tomography (PET) imaging is a methodology of molecular imaging based on nuclear imaging. PET imaging in combination with radiolabeled TKIs improves accuracy of quantitative imaging strategies and the probability of successful drug development, and may facilitate the stratification of patients. Here, we describe a protocol for PET imaging using radiolabeled TKI in preclinical trials.

Abstract The recent breakthrough identifying the onco-driver fusion mutant of ALK kinase and its inhibitor crizotinib commercially termed as Xalkori has brought significant benefit to a portion of non-small cell lung cancer (NSCLC) patients. However, a number of clinical issues in ALK-positive lung cancer remain, involving resistance to crizotinib caused by secondary mutation, amplification of the ALK gene, activation of alternative pathways, and metastatic resistance, etc. Among these mechanisms, brain metastasis is a critical issue because of its poor prognosis. We have recently identified ASP3026 as a novel type of ALK inhibitor under development, and have reported that ASP3026 shows antitumor activities in several crizotinib-refractory models including gate keeper mutants. Here, we report the first PET imaging of an ALK inhibitor using [11C]ASP3026. The study has revealed that ASP3026 shows a brain tumor permeability in an intracranial xenograft model of H2228-luc ALK fusion positive cells. In this model, significant growth inhibition of H2228 intracranial tumor was observed by treatment with ASP3026 (10 mg/kg, q.d.), but not with crizotinib (10 mg/kg, q.d.) as determined by a bioluminescent imaging technique. Pharmacokinetic measurements of ASP3026 and crizotinib indicated that ASP3026 showed a four-fold better brain penetration than crizotinib on AUC0-24 base analysis, with a brain/plasma ratio=0.72 and 0.18 for ASP3026 and crizotinib, respectively. Further, we synthesized positron-labeled [11C]ASP3026 and performed PET imaging to clarify penetration of ASP3026 into cranial tumors. Quantitative analysis of [11C]ASP3026-PET data indicated that ASP3026 showed higher uptake into cranial tumors (SUV=3.0) than brain parenchyma (SUV=0.8). Moreover, comparison of pharmacokinetic profiles in several tumor models showed that tumor uptake of ASP3026 was higher than that of surrounding tissue, suggesting that tumor accumulation of ASP3026 was dependent on the microenvironment of tumor. Taken together, these results suggest that ASP3026 has favorable properties that may be useful for the treatment of brain metastases in ALK-positive NSCLC patients. Thus, PET imaging using 11C-labeled ASP3026 may allow the tumor penetration of ASP3026 to be clarified in any primary or metastatic tumor site. Citation Format: Hiroshi Fushiki, Rika Saito, Makoto Jitsuoka, Itsuro Shimada, Yutaka Kondoh, Hideki Sakagami, Yukiko Funatsu, Akihiro Noda, Yoshihiro Murakami, Sousuke Miyoshi, Yoko Ueno, Satoshi Konagai, Takatoshi Soga, Shintaro Nishimura, Masamichi Mori, Sadao Kuromitsu. First demonstration of in vivo PET imaging for ALK inhibitor using [11C]ASP3026, a novel brain-permeable type of ALK inhibitor. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2678. doi:10.1158/1538-7445.AM2013-2678

Abstract EML4-ALK translocation has been validated as a therapeutic target in a subset of non-small cell lung cancer (NSCLC) patients. Crizotinib, an FDA-approved ALK inhibitor, is effective against several types of human cancers with ALK abnormalities including EML4-ALK, RANBP2-ALK and ALK mutations. However, long-term treatment is often limited by the development of resistant tumors and distant metastases. Multiple brain metastases are a critical issue because of their poor prognosis despite the standard radiotherapy. The reason for metastases and development of tumor in the brain could be attributable to the poor penetration of crizotinib into central nervous system, as previously reported. ASP3026 is a selective ALK inhibitor which shows antitumor activities in several crizotinib-refractory models including gate keeper mutants. Here, we established an intracranial xenograft model by implanting NCI-H2228 cells directly in the brain of immunocompromised mice. Xenografted cells develop intracranial tumors which grow in murine brain and eventually become lethal, resembling the clinical tumors that metastasized in the brain. To determine the anti-tumor activity against tumors in the brain, mice were treated with ASP3026 or crizotinib for 2 weeks and held to observe survival duration. Ten, 30 and 100 mg/kg daily oral administration of ASP3026 dose dependently inhibited tumor growth in brain, and tumor regression was achieved in 30 and 100 mg/kg groups confirmed by MR imaging. These results were supported by the observation that Ktrans and IAUC (90s) were significantly decreased in 30 and 100 mg/kg groups using dynamic contrast enhanced MR imaging (DCE-MRI). Furthermore, ASP3026 significantly prolonged the survival of tumor bearing mice compared to vehicle treatment group. On the other hand, 30mg/kg daily oral administration of crizotinib inhibited the tumor growth during the treatment period but did not produce significant survival benefit. Taken together, these results indicate that ASP3026 shows better efficacy than crizotinib and improves overall survival in an intracranial mouse xenograft model, suggesting that ASP3026 may have potential to benefit EML4-ALK positive NSCLC patients with brain metastases. Citation Format: Satoshi Konagai, Hiroshi Fushiki, Hideki Sakagami, Yoko Ueno, Masamichi Mori, Itsuro Shimada, Yutaka Kondoh, Sousuke Miyoshi, Shintaro Nishimura, Sadao Kuromitsu. ASP3026, a selective ALK inhibitor, shows anti-tumor activity in a mouse model xenografted with NCI-H2228 intracranially. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 918. doi:10.1158/1538-7445.AM2013-918

The development of positron-emission tomography (PET) and X-ray computed tomography (CT) imaging has improved the detection of tumor burden and, in turn, pre-clinical drug development and clinical treatment. In pre-clinical drug development, clinically-relevant murine cancer models, such as orthotopic models of lung cancer, have provided an accurate representation of tumor burden in humans. However, evidence demonstrating the capability of imaging-guided evaluation of these clinically-relevant models is limited. Here, we combined (18)F-fluorothymidine (FLT)-PET/CT imaging and a murine model of human non-small cell lung cancer (NSCLC) to improve the accuracy of anticancer drug evaluation in pre-clinical studies. We found that FLT-PET/CT imaging enabled the progression of pulmonary tumors to be longitudinally monitored rather than FDG-PET/CT. Furthermore, in an efficacy study of a standard treatment of docetaxel in a murine lung cancer model, FLT-PET imaging detected the anticancer response earlier than volumetric analysis by CT imaging. We, thus, observed a relationship between the alteration of FLT signals and Ki-67 index in the pulmonary tumor during the period of chemotherapy. These results indicate that the combination of FLT-PET/CT imaging and an orthotopic NSCLC model is an effective strategy for evaluating clinical efficacy and potential of an anticancer agent during pre-clinical development.

Introduction: Sepantronium bromide (YM155) is an antitumor drug in development and is a first-in-class chemical entity, which is a survivin suppressant. We developed a radiosynthesis of [C-11]YM155 to non-invasively evaluate its tissue and tumor distribution in mice bearing human prostate tumor xenografts.Methods: Methods utilizing [C-11]acetyl chloride and [C-11]methyl triflate, both accessible with automated radiosynthesis boxes, were evaluated. The O-methylation of ethanolamine-alkolate with [C-11]methyl triflate proved to be the key development toward a rapid and efficient process. The whole-body distribution of [C-11] YM155 in PC-3 xenografted mice was examined using a planar positron imaging system (PPIS).Results: Sufficient quantities of radiopharmaceutical grade [C-11]YM155 were produced for our PET imaging and distribution studies. The decay corrected (EOB) radiochemical yield was 16-22%, within a synthesis time of 47 min. The radiochemical purity was higher than 99%, and the specific activity was 29-60 GBq/mu mol (EOS). High uptake levels of radioactivity (%ID/g, mean +/- SE) were observed in tumor (0.0613 +/- 0.0056), kidneys (0.0513 +/- 0.0092), liver (0.0368 +/- 0.0043) and cecum (0.0623 +/- 0.0070). The highest tumor uptake was observed at an early time point (from 10 min after) following injection. Tumor-to-blood and tumor-to-muscle uptake ratios of [C-11]YM155, at 40 min after injection, were 26.5 (+/-2.9) and 25.6 (+/-3.6), respectively.Conclusion: A rapid method for producing a radiopharmaceutical grade [C-11]YM155 was developed. An in vivo distribution study using PPIS showed high uptake of [C-11]YM155 in tumor tissue. Our methodology may facilitate the evaluation and prediction of response to YM155, when given as an anti-cancer agent. (C) 2013 Elsevier Inc. All rights reserved.

We constructed a system for safety navigation using a System Dynamics computer simulation.The algorithm for judgment of searching route includes “The International Regulations for Preventing Collisions at Sea (COLREGs)”. And the system of this research includes several factors that used by ships' officers, such as “Bumper Model” and “CPA and TCPA”.In most of existing systems, the route is analyzed by moving on inflexible positions which were defined by cells or nodes beforehand. In the system constructed, it's possible to analyze high precise route searching by defining ships' positions with longitude-and-latitude. And the system can analyze with high precision routes by referring ship specifications.In this research compare route searched by the system with actually route that extract from AIS record.We evaluated risk of each route using “SJ Values (Subjective Judgment Value)” and reached the conclusion that the system can search safer route than actually route.

ポジトロン断層撮像法（PET：positron emission tomography）は，医療現場でがんや脳機能等の診断に使われている短半減期の放射性診断薬を用いた核医学診断技術である．動物でも測定可能なことから，非臨床と臨床データをブリッジできるトランスレーショナル研究の有力なツールとして注目されている．創薬への利用としてはターゲット臓器への到達薬物量の定量（薬物動態試験）や，受容体占有率試験による有効投与量の推定・POM（proof of mechanism）の取得（薬理試験），開発候補品の他剤との比較・差別化等に利用可能であり，最近ではさらに安全性試験やコンパニオン診断薬への応用も検討され始めている．当社では長年国内外のアカデミア，異業種とのオープンイノベーションや国家プロジェクトを通じてPETの要素技術開発や人材育成に取り組み，国内製薬初の自社PETイメージング研究施設と組織を構築して研究・開発を推進中である．本稿ではPETの創薬応用に向けた当社の取り組みについて概説する．

Introduction: Telmisartan is a widely used, long-acting antihypertensive agent. Known to be a selective angiotensin II type 1 (AT(1)) receptor (AT(1)R) blocker (ARB), telmisartan acts as a partial agonist of peroxisome proliferator-activated receptor-gamma (PPAR-gamma) and inhibits centrally mediated effects of angiotensin II in rats following peripheral administration, although the brain penetration of telmisartan remains unclear. We investigated the brain concentration and localization of telmisartan using C-11-labeled telmisartan and positron emission tomography (PET) in conscious rhesus macaques.Methods: Three male rhesus macaques were bolus intravenously administered [C-11]telmisartan either alone or as a mixture with unlabeled telmisartan (1 mg/kg). Dynamic PET images were acquired for 95 min following administration. Blood samples were collected for the analysis of plasma concentration and metabolites, and brain and plasma concentrations were calculated from detected radioactivity using the specific activity of the administered drug preparation, in which whole blood radioactivity was used for the correction of intravascular blood radioactivity in brain.Results: Telmisartan penetrated into the brain little but enough to block AT(1)R and showed a consistently increasing brain/plasma ratio within the PET scanning period, suggesting slow clearance of the compound from the brain compared to the plasma clearance. Brain/plasma ratios at 30, 60, and 90 min were 0.06, 0.13, and 0.18, respectively. No marked localization according to the AT(1)R distribution was noted over the entire brain, even on tracer alone dosing.Conclusions: Telmisartan penetrated into the brain enough to block AT(1)R and showed a slow clearance from the brain in conscious rhesus macaques, supporting the long-acting and central responses of telmisartan as a unique property among ARBs. (C) 2012 Elsevier Inc. All rights reserved.

The use of radioisotopes is important in pharmaceutical research and development (R&D). They are frequently used in non-clinical and clinical studies for the development of compounds for different therapeutic areas, such as central nervous system (CNS) diseases (e.g. Dementia, Alzheimer’s disease (AD), and Parkinson’s disease), oncology, and metabolic diseases (e.g. diabetes mellitus). Pharmaceutical companies invest a lot of time and money in research on new treatment strategies for diseases with a high medical need, such as oncology and metabolic diseases. A large amount of drugs fails during development due to toxicity and/or the lack of efficacy (Kola, I, 2008). Several attempts are being made to improve this, such as obtaining a better understanding of the pathophysiology of diseases, development of robust animal models, the application of biomarkers, development of pharmacokinetic (PK) pharmacodynamic (PD) models, and the application of non-invasive techniques such as positron emission tomography (PET) in an early stage of development.

Objectives F-18-FDG PET with voxel-based statistical image analysis plays an important role in the diagnosis of Alzheimer's disease (AD). However, the effect of an age-matched and sex-matched or mismatched normal database (NDB) on the diagnostic performance of F-18-FDG PET has not yet been investigated systematically. The aim of this study was to determine whether an age-matched and sex-matched NDB is necessary for the detection of AD using F-18-FDG PET.Methods We generated 11 NDB sets for F-18-FDG PET, including six age-specific NDB sets consisting of participants ranging in age from 20 to 70 years, one age-non-specific NDB set, one age-matched NDB set, two sex-specific NDB sets, each consisting of 20 men or 20 women, and one sex-matched NDB set. The average z-scores in predefined AD-specific regions of interest of the PET images were calculated using those NDB sets and a receiver-operating characteristic analysis was carried out to assess the diagnostic performance of F-18-FDG PET to discriminate 46 patients with AD from 50 normal controls.Results There was no significant difference in each area under the receiver-operating characteristic curve using either age-matched/mismatched NDB sets or sex-matched/mismatched NDB sets.Conclusion The diagnostic performance of F-18-FDG PET was rather insensitive to differences in age or sex in the NDB, indicating that exact age-matched or sex-matched NDB may not be essential for discriminating patients with AD from normal participants using F-18-FDG PET. Nucl Med Commun 32: 1128-1133 (C) 2011 Wolters Kluwer Health vertical bar Lippincott Williams & Wilkins. Nuclear Medicine Communications 2011, 32: 1128-1133

Oligopeptide transporters are abundantly expressed in various types of cancer cells. We here synthesized two novel dipeptides, l-phenylalanyl sarcosine (Phe-Sar) and 4-(4-methoxyphenyl)-l-phenylalanyl sarcosine (Bip(OMe)-Sar), and examined their effect on the growth of human pancreatic cancer AsPC-1 cells, which are known to highly express oligopeptide transporter PEPT1/SLC15A1. Growth of AsPC-1 cells was inhibited by these two peptides and a typical PEPT1/SLC15A1 substrate Gly-Sar. Growth inhibition by Gly-Sar, Phe-Sar and Bip(OMe)-Sar was concentration-dependent with half-maximal inhibitory concentration of 50, 0.91 and 0.55mM, respectively. These peptides also inhibited PEPT1-mediated [(3)H]Gly-Sar uptake with half-maximal inhibitory concentration of 2.6, 0.81 and 0.27mM, respectively. Thus, the rank order of the tumor cell growth inhibition by these three peptides was the same as that of PEPT1-inhibitory activity. Growth of AsPC-1 cells was also inhibited by 2-aminobicyclo(2,2,1)heptane-2-carboxylic acid (BCH), which is a typical inhibitor of amino acid transporter system L. The growth inhibition by BCH and Gly-Sar was additive, suggesting that these compounds act at distinct loci. Oligopeptide transporters thus appear to be a promising target for inhibition of pancreatic cancer progression. These results also proposed the idea that oligopeptide transporter is required for growth of AsPC-1 cells.

The recent development in radiosynthesis of the 11C-carbamate function increases the potential of [11C]GR103545, which for the last decade has been regarded as promising for imaging the kappa-opioid receptor (κ-OR) with PET. In the present study, [11C]GR103545 was evaluated in awake rhesus macaques. Separate investigations were performed to clarify the OR subtype selectivity of this compound.

8087 Background: YM155 is a novel small-molecule survivin suppressant, and is currently under development for the treatment of non- Hodgkin's lymphoma in a phase II clinical trial. Therapy monitoring by non-invasive PET (positron emission tomography) imaging study allow a straightforward translation from preclinical to clinical research. The aim of this study was to evaluate the utility of the PET tracers, FLT(3'-deoxy-3'-[18F]fluorothymidine) and FDG(2-[18F]fluoro-2-deoxy-D-glucose) to monitor tumor response to YM155 in combination with rituximab (RTX) treatment in diffuse large B cell lymphoma (DLBCL) xenograft model. Methods: YM155 (0.5 mg/kg/day) was administered on day 0 (D0) as a 7-day continuous subcutaneous infusion, and RTX (50 mg/kg/day) was administered by intravenous bolus injection on D0 and D+2 in mice (N=4) with the human derived DLBCL xenograft, WSU-DLCL- 2. FLT/FDG uptake was evaluated using a small-animal PET scan, Inveon (Siemens), on D0 (before treatment), D+3, +7, and +14 / on D0 and D+3 after treatment, respectively. At each time point, excised tumors from other mice (N=4) were evaluated for histopathology with proliferative activity (Ki-67 index). Results: YM155 in combination with RTX (combo) resulted in significant (P <.05) regression of tumor size compared to the RTX monotherapy group without loss of body weight. PET imaging data showed tumor uptake of FLT/FDG significantly decreased in the YM155 alone (-17%/-18%) and combo (-50%/-42%) groups on D+3 compared to baseline (D0) in contrast to the RTX alone (+72%/+12%) or saline (+126%/+48%) groups where uptake increased, respectively. In addition, significant (P <.05) decrease of the Ki-67 positive cells in the tumor compared to the RTX alone group could be observed on D+3. Conclusions: YM155 may increase anti-tumor activity of RTX for the treatment of DLBCL. FLT-PET as a cell proliferation imaging, FDG-PET imaging reflecting glucose metabolism and Ki-67 index, respectively may be a useful biomarker of this combination therapy. No significant financial relationships to disclose.

We present a case of gastric ulcer with bleeding following chemotherapy with rituximab for non-Hodgkin’s lymphoma (NHL).

[18F]Fluoro-2-deoxyglucose positron emission tomography (FDG-PET) is a useful tool for measuring the regional cerebral metabolic rate of glucose (rCMRglu), which is an index of neuronal activity. Donepezil, an acetylcholine esterase inhibitor (AChEI), has been recommended as a treatment option for patients with Alzheimer’s disease (AD). We aimed to characterize the effects of donepezil on rCMRglu using FDG-PET in non-human primates.