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Background: The application of personalized cancer treatment based on genetic information and surgical samples has begun in the field of cancer medicine. However, biopsy may be painful for patients with advanced disease that do not qualify for surgical resection. Patient-derived xenografts (PDXs) are cancer models in which patient samples are transplanted into immunodeficient mice. PDXs are expected to be useful for personalized medicine. The aim of this study was to establish a PDX from body fluid (PDX-BF), such as peritoneal and pleural effusion samples,to provide personalized medicine without surgery. Methods PDXs-BF were created from patients with ovarian cancer who had positive cytology findings based on peritoneal and pleural effusion samples. PDXs were also prepared from each primary tumor. The pathological findings based on immunohistochemistry were compared between the primary tumor, PDX, and PDX-BF. Further, genomic profiles and gene expression were evaluated using DNA and RNA sequencing to compare primary tumors, PDXs, and PDX-BF. Results Among the 15 patients, PDX-BF was established for 8 patients (5 high-grade serous carcinoma, 1 carcinosarcoma, 1 low-grade serous carcinoma, and 1 clear cell carcinoma); the successrate was 53%. Histologically, PDXs-BF have features similar to those of primary tumors and PDXs. In particular, PDXs-BF had similar gene mutations and expression patterns to primary tumors and PDXs. Conclusions PDX-BF reproduced primary tumors in terms of pathological features and genomic profiles, including gene mutation and expression. Thus, PDX-BF may be a potential alternative to surgical resection for patients with advanced disease.

This study examined the effects of the presence and absence of Vitamin C (VC) intake and exercise on alveoli and cytokines in mice.

Cesarean section (CS), the rate of which is increasing worldwide, may be associated with complications. Although pelvic abscess after CS is rare, it is difficult to treat. We herein report two cases of pelvic abscess treated laparoscopically after CS. The abscesses of the patients were located in the pouch of Douglas and the uterine scar after CS, respectively. Several days after CS, the patients presented with lower abdominal pain and fever. Laparoscopic drainage was performed because imaging revealed a pelvic abscess that was not amenable to drainage through interventional radiology. The patients recovered from infection and were discharged four days after drainage.

[Purpose] It has been reported that exercise affects skeletal muscle in the chronic obstructive pulmonary disease (COPD) disease model. In this study, we examined the effects of neuromuscular electrical stimulation (NMES) in skeletal muscle on alveoli and cytokines. [Materials and Methods] We used twenty wild-type mice, randomly divided into three groups: Group A: Control (non-COPD, non-amyotrophia, non-NMES), Group B: COPD, amyotrophia with NMES and Group C: COPD, amyotrophia without NMES. Among those, a group of mice with ages from 12 to 14 weeks were used to create a chronic obstructive pulmonary disease (COPD) model, a group of mice with ages from 15 to 16 weeks was used to create a disuse syndrome by hind limb suspension, and a group of mice with ages from 17 to 28 weeks (12 weeks) were used to implement NMES. In this study, we used the real-time PCR method to assess the mRNA expression levels. We also conducted morphological analysis, assessed macrophage expression level by staining (general staining and immunostaining), and employed spirometry. [Results] Our study results showed significant decreases in Interleukin-6 (IL-6) levels in the lungs and muscle RING-finger protein-1 (MuRF1) in the muscles. Moreover, the pulmonary stromal macrophage marker (F4/80) and the protease marker (MMP12) showed significantly decreased expression, while no change was observed in the morphological of the alveolar spaces (mean linear intercept). [Conclusion] On the basis of these findings, our study reveals that NMES affects cytokines and macrophages in COPD skeletal muscle atrophy.

The number of heart failure patients is increasing rapidly in Japan because of its large elderly population. As age increases, arterial stiffness and physical dysfunction progress. This study aimed to evaluate the association between the physical function and arterial stiffness in elderly heart failure patients.This retrospective, observational study includes data from 100 heart failure patients aged ≥ 65 years who were admitted to our hospital and underwent cardiac rehabilitation. The Cardio-Ankle Vascular Index (CAVI) was measured as an indicator of arterial stiffness. Body composition was assessed by bioelectrical impedance analysis. To determine the degree of physical function, we assessed handgrip strength, five-meter walk speed (5MWS), five-repetition sit-to-stand time (5RSST) and six-minute walk distance (6MWD). Sarcopenia was defined using Asian guidelines based on physical function and body composition.Among 100 patients, 47.0% of patients had sarcopenia. After adjustments for age, sex, atrial fibrillation, and ischemic cardiomyopathy, CAVI was significantly higher in with sarcopenia patients than those without sarcopenia. Age, handgrip strength, 5MWS, 5RSST, and 6MWD were associated with CAVI, and 6MWD was as an independent determinant factor of CAVI.6MWD was recognized as an accurate physical function indicator. These findings suggested that physical function and arterial stiffness complement each other. To restore cardiac dysfunction, improving both arterial stiffness and physical function might be useful.

We performed a comprehensive quantification of 20 amino acids in RPMI 1640 medium-cultured human colorectal adenocarcinoma cells to evaluate the efficacy of 5-fluorouracil treatment under hypoxic and hypoglycemic conditions, which mimic the tumor microenvironment. In this study, we developed a simple and comprehensive analytical method by using LC-MS/MS connected to the Intrada amino acid column, which eluted amino acids within 9 min. The present method covered a linearity range of 3.6 - 1818 μM, except for Gly (227 - 1818 μM), Ala, Asp, His (7.1 - 1818 μM each), and Trp (3.6 - 909 μM). The limits of detection were in the range of 0.02 - 38.0 pmol per injection in a standard solution. Amino acid concentration data were analyzed using principal-component analysis to represent samples on two-dimensional graphs. Linear discriminant analysis was used to classify samples on the score plots. Using this approach, the effect of 5-fluorouracil treatment could be successfully discriminated at high discrimination rates. Moreover, several amino acids were extracted from corresponding loading plots as candidate markers for distinguishing the effects of the 5-fluorouracil treatment or tumor microenvironmental conditions. These results suggest that our proposed method might be a useful tool for evaluating the efficacy of anticancer drugs in the tumor microenvironment.

Objectives No established approaches exist for the pharmacological management of cardiovascular diseases (CVDs) in residents of long-term care facilities (LTCFs). This study aimed to evaluate the use of drugs for CVD prevention and treatment (CVD-related drugs) in a major type of LTCF in Japan. Methods This study included 1318 randomly selected residents at 349 intermediate care facilities for older adults (called Roken). Prescriptions were investigated at admission and two months after admission according to therapeutic categories. Logistic regression was used to identify residents’ characteristics that were associated with prescriptions of CVD-related drugs. Results Prescriptions of all types of drugs and CVD-related drugs decreased in 36 % and 16 % of residents, respectively. Half of the residents received antihypertensives, a quarter received antiplatelets and diuretics, whereas one-tenth received antidiabetics, oral anticoagulants, and lipid-modifying drugs. The prevalence of most of individual drug categories were similar among residents with different physical or cognitive function, except for fewer antihypertensive and lipid-modifying drugs in those with severe cognitive disability. Adjusted analyses for prescriptions at two months after admission revealed that bedridden residents were more likely to be prescribed diuretics but less likely to be prescribed antihypertensives, antiplatelets, or lipid-modifying drugs. Residents with severe cognitive disability were less likely to be prescribed antihypertensives or lipid-modifying drugs. A known history of cardiovascular events was associated with greater use of CVD-related drugs. Conclusion CVD-related drugs were commonly prescribed for Roken residents, including those with low physical and cognitive functions. Deprescribing may contribute to the optimization of pharmacotherapy in LTCF residents.

An approach to spiropyrazole derivatives containing iminolactone and/or cyclic imide moiety starting from 1H-pyrazole-4-acetic acid derivative is described. Hydrolysis of C-cyanomethylated 1H-pyrazole-4-acetic acid methyl ester (), which was easily prepared from 1H-pyrazole-4-acetic acid derivative by a C-cyanomethylation, led to the C-cyanomethylated 1H-pyrazole-4-acetic acid (). Compound was reacted with ethanol in the presence of tin(IV) chloride in refluxing chloroform to give the key intermediate ethyl imidate (). Sodium hydride-assisted lactonization of in N,N-dimethylformamide afforded the spiropyrazole derivative containing iminolactone moiety (). On the other hand, thermal treatment of with sodium acetate in the absence of solvent caused another intramolecular cyclization to yield the spiropyrazole derivative containing cyclic imide moiety ().

e17041 Background: G protein-coupled receptor 30 (GPR30) is a 7-transmembrane estrogen receptor that functions alongside traditional estrogen receptors to regulate the cellular responses to estrogen. Recent studies suggest that the high expression of GPR30 is associated with a poor prognosis in breast cancer or endometrial cancer. Although the role of GPR30 in ovarian cancer was unclear, we revealed that GPR30 is associated with poor prognosis in ovarian cancer. On the other hand, Epithelial-to-Mesenchymal Transition (EMT) is involved in cancer metastasis. The purpose of this study is to reveal how GPR30 was associated with poor prognosis and whether associated with EMT in ovarian cancer. Methods: We examined whether GPR30 signaling activates the EGFR-Akt pathway in an ovarian cancer cell line (Caov-3) by a Western blotting analysis. We also examined the effect of GPR30 on EMT were evaluated in Caov-3, which were cultured both in two-dimensional (2D) culture and three-dimensional (3D) culture model. GPR30 agonist, G1, was used to confirm the regulatory effects of GPR30 on the change of phenotypic modulation and EMT markers expression. Results: The phosphorylation of the EGFR and Akt could be significantly enhanced by G1 (p < 0.05) and inhibited by a Src family kinase inhibitor. In 3D culture, the stimulation of GPR30 leads the floating and sphere formation in Caov-3. G1-induced EMT was observed with related regulation of EMT markers expression at both mRNA and protein level. G1 induced the decrease of E-cadherin level and the increase of Snail and Vimentin in RT-PCR and Western blotting. Knockdown of GPR30, using siRNA, blocked G1-induced EMT. Conclusions: GPR30 increases the phosphorylation of Akt via the EGFR in ovarian cancer cells and changes ovarian cancer cells to the EMT state.GPR30 might be an important molecule related to metastasis process in ovarian cancer.

In this study with 30 nurses as study subjects, we examined how 12 weeks of a study treatment affected body measurements, blood biochemistry, and the oxidative stress regulatory system. Subjects were randomly divided into two groups: one that received licorice flavonoid oil (LFO) capsules and l-carnitine capsules (supplement group n = 15), and another that received placebo capsules (placebo group n = 15), for 12 consecutive weeks. Measurements of oxidative stresses and other tests were performed at baseline and after 4, 8, and 12 weeks of treatment. Results showed that only the supplement group showed a significant decrease in waist circumference at 8 weeks when compared to baseline, while neither group showed a change in body mass index. No changes in blood biochemistry were observed in either group. Only the supplement group showed a significant decrease in oxidative stresses at 8 weeks when compared to baseline, while the placebo group did not show any change. Both groups showed a significant decrease in anti-oxidant potential, but at 12 weeks the supplement group showed a significantly higher level than the placebo group. While the supplement group showed a stable BAP to d-ROM ratio (representing latent anti-oxidant potential), the placebo group had a significant decrease. These results demonstrate that 12 weeks of LFO and l-carnitine consumption decreases waist circumference and oxidative stresses and maintains latent anti-oxidant potential.

5'-AMP-activated protein kinase (AMPK) and its pharmacological modulators have been targeted for treating type 2 diabetes. Extracellular uridine 5'-diphosphate (UDP) activates P2Y6 receptors (P2Y6Rs) in pancreatic β-cells to release insulin and reduce apoptosis, which would benefit diabetes. Here, we studied the role of P2Y6R in activation of AMPK in MIN6 mouse pancreatic β-cells and insulin secretion. Treatment with a potent P2Y6R dinucleotide agonist MRS2957 (500nM) activated AMPK, which was blocked by P2Y6R-selective antagonist MRS2578. Also, MRS2957 induced phosphorylation of acetyl-coenzyme A carboxylase (ACC), a marker of AMPK activity. Calcium chelator BAPTA-AM, calmodulin-dependent protein kinase kinase (CaMKK) inhibitor STO-069 and IP3 receptor antagonist 2-APB attenuated P2Y6R-mediated AMPK phosphorylation revealing involvement of intracellular Ca(2+) pathways. P2Y6R agonist induced insulin secretion at high glucose, which was reduced by AMPK siRNA. Thus, P2Y6R has a crucial role in β-cell function, suggesting its potential as a therapeutic target in diabetes.

[Purpose] Hormone replacement therapy has been reported to be effective for alleviating menopausal symptoms, its side effects have been a concern. Therefore, it is necessary to investigate methods that could alleviate menopausal symptoms but with fewer side effects. Few previous reports have investigated the effects of simplified, viable manual lymph drainage, particularly the effects of one-time therapy on physiologically active substances and other variables. Effects of one-time simplified lymph drainage performed at salon A were investigated in females with any type of menopausal symptoms, such as edema or a sense of fatigue. [Subjects and Methods] Before and after lymph drainage, saliva was collected to detect substances that would reveal immune function. Questionnaire surveys were also conducted before and after therapy. [Results] Cortisol and dehydroepiandrosterone levels were significantly reduced after therapy compared with those before therapy. The questionnaire survey showed a significant decrease in the visual analog scale. Also apparent were positive opinions, such as "I feel better" and "My legs feel lighter." [Conclusion] These results demonstrated the stress reduction effect of one-time simplified lymph drainage, which decreased cortisol and dehydroepiandrosterone levels. This therapy was also shown to produce positive mental and physical effects.

As a part of systematic investigation of synthesis and biologically active compounds of pyrazole derivatives containing transition metal, several new pyrazole copper(II) complexes 3a−f were synthesized from pyrazole sodium salts 2a−f, which were produced from spiro-pyrazoles 1a−f and sodium hydride by a ring-opening reaction. All the synthesized compounds were characterized by spectroscopic analysis. Pyrazole copper(II) complexes 3a−d and 3f exhibited high DNA cleavage activity in vitro. Furthermore, compounds 3a−f were tested for their growth inhibitory activity in A549 lung cancer, B16F10 murine melanoma, and HeLa human uterine carcinoma cells. Compounds 3c,d displayed moderate B16F10 and HeLa inhibitory activity levels (3c: IC50 = 45 μM in B16F10 cells and 34 μM in HeLa cells, 3d: IC50 = 50 μM in B16F10 cells and 32 μM in HeLa cells).

A novel synthetic route to 4-pyridazineacetic acids 10–12 has been achieved by the ring-expansion reaction of N-cyanomethylated 3-pyrazoline-4-acetic acids 7–9. 1H-Pyrazole-4-acetic acids 1–3 were reacted with iodoacetonitrile in the presence of triethylamine in refluxing acetonitrile to give the corresponding C-cyanomethylated 1H-pyrazole-4-acetic acids 4–6 as major products together with N-cyanomethylated 3-pyrazoline-4-acetic acids 7 and 8 as minor products. On the other hand, reactions of 1 and 3 with chloroacetonitrile in the presence of triethylamine in refluxing chloroform afforded the corresponding N-cyanomethylated 3-pyrazoline-4-acetic acids 7 and 9 as major products. Thermal treatment of 7–9 with sodium hydride in N,N-dimethylformamide caused ring expansion to yield the corresponding 4-pyridazineacetic acids 10–12.

[Purpose] The effects of exercise and food consumption on the oxidative stress regulation system were studied using 45 male ICR mice. [Subjects and Methods] The mice were randomly divided into two groups: a control group (CO group) and a group consuming a reduced form of coenzyme Q10 (H2CoQ10: QH) (QH group). Both groups were made to run on a treadmill for animals (TM) twice and the changes in their running time were measured. For the oxidative stress regulation system, plasma oxidative stresses (d-ROM test) and plasma anti-oxidant potential (BAP test) were measured before and after treadmill running with analytical equipment for reactive oxygen and free radicals, and then the BAP/d-ROM ratio was calculated. [Results] The measurements and calculations showed a significant increase in the running time of the QH group. With respect to the oxidative stress regulation system, no significant change was observed in the BAP/d-ROM ratio. [Conclusion] These results showed that a single consumption of QH produced an effect on exercise, but had no effect on the oxidative stress regulation system.

As a part of systematic investigation of synthesis and biologically active compounds of thiazolidine (TZD) derivatives containing pyrazole ring system, several new pyrazole–TZD derivatives and have been synthesized. Compounds were prepared from N-substituted TZDs and 1H-pyrazole-4-carboxaldehyde by Knoevenagel-type reaction. Treatment of with sodium hydride at room temperature caused dimerization reaction to afford the corresponding spirocompounds . All the synthesized compounds were characterized by spectroscopic analysis. In vitro, the synthesized compounds and were tested for their growth inhibitory activity in A549 lung cancer, B16F10 murine melanoma, and HeLa human uterine carcinoma cells and for their differentiation of 3T3-L1 preadipocytes to adipocytes. The results showed that compound possessed growth inhibitory effect of B16F10 cells (IC50 = 27 μM) and compounds had induction effect on the differentiation of 3T3-L1 preadipocytes. J. Heterocyclic Chem., (2011).

A chymase inhibitor SUN13834 has been shown to improve skin condition in animal models for atopic dermatitis. In the present study, effective dosages of SUN13834 for atopic dermatitis patients were predicted by pharmacokinetic/pharmacodynamic (PK/PD) analyses of SUN13834 in NC/Nga mice, which spontaneously develop atopic dermatitis-like skin lesions. For the PK/PD analyses, we utilized the minimum effective plasma concentration of unbound SUN13834 in late-phase reaction of trinitrochlorobenzene (TNCB)-induced biphasic dermatitis in mice, based on the assumption that the minimum effective plasma concentrations are the same among the two animal models. In late-phase reaction of biphasic dermatitis, SUN13834 was most effective when its plasma concentration was highest at the elicitation, and the minimum effective plasma concentration of unbound SUN13834 at the elicitation was calculated to be 0.13-0.2 ng/mL. Oral administration of SUN13834 improved dermatitis in NC/Nga mice at 15 mg/kg (twice a day; bid) and 30 mg/kg (once a day; qd), but not at 60 mg/kg (every other day; eod). At the three dosages, the duration times over the plasma level of 0.13-0.2 ng/mL were 16.1-20.3, 10.7-12.2 and 7.8-8.8h, respectively, suggesting an importance of maintenance of the minimum effective plasma concentration for at least about 10-12h. The clinical effective dosage predicted in this paper is also discussed in relation to a recently conducted Phase 2a study.

Dihydropyridazinones 4a, 4b, N-substituted dihydropyrazoles 5b, 5c, 5d, and O-substituted pyrazoles 6a, 6b, 6c, 6d have been synthesized starting from spirocyclopropanepyrazole derivative 2. Treatment of 2 with α-chloro esters, e.g., methyl chloroacetate, ethyl chloroacetate, isopropyl chloroacetate, and tert-butyl chloroacetate, in potassium carbonate/sodium iodide system caused ring opening and subsequent C- or N-attack nucleophilic substitution to give the corresponding dihydropyridazinones 4a, 4b and N-substituted dihydropyrazoles 5b, 5c, 5d. On the other hand, in the absence of sodium iodide, O-substituted pyrazoles 6a, 6b, 6c, 6d were obtained from 2 via an O-attack nucleophilic substitution. J. Heterocyclic Chem., 2011.

An approach to pyrano[2,3-c]pyrazoles starting from spirocyclopropanepyrazoles via a ring-opening/cyanomethylation and intramolecular cyclization is described. Reactions of spirocyclopropanepyrazoles with chloroacetonitrile in the presence of sodium hydride gave the corresponding cyanomethoxypyrazoles . Treatment of with sodium hydride at room temperature caused intramolecular Michael addition reaction to afford the corresponding pyrano[2,3-c]pyrazoles . J. Heterocyclic Chem., (2009).

A series of new spiro[cyclopropane-1,4′-pyrazol-3-one] derivatives 3a-h were synthesized by the reaction of 4-arylidene-3H-pyrazol-3-one 1 with secondary and tertiary carbanions derived from a methylene and methine group bearing both a leaving group and electron-withdrawing group, e.g. methyl chloroacetate, ethyl chloroacetate, isopropyl chloroacetate, tert-butyl chloroacetate, chloroacetonitrile, 2-chloro-N,N-diethylacetamide, methyl 2-chloropropionate and 2-chloropropionitrile, in the presence of sodium hydride. All the synthesized compounds 3a-h were active against Candida albicans with MIC ≤ 25 μg/mL in vitro.

Two methods for the preparation of N-methyl-4-(methylthio)thieno[2,3-d]pyrimidinium salts 6a,b and 13a,b are described. Treatment of 6a,b and/or 13a,b with active methylene compounds such as malononitrile and ethyl cyanoacetate in the presence of sodium methoxide caused nucleophilic addition followed by elimination of methanethiol, giving the corresponding N-methyl-4-ylidenethieno[2,3-d]-pyrimidines 7a,b, 8a,b, 14a,b and 15a,b.

The one-pot synthesis of 4-alkoxy-5,6-dihydro-furo(and -thieno)[2,3-d]pyrimidines is described. The reactions of 2-benzamido-4,5-dihydro-3-furan(and -3-thiophene)carbonitriles 1a-d and 2a-c with ethanol and/or methanol in the presence of zinc chloride and triethylamine gave the corresponding 4-alkoxy-5,6-dihydro-furo(and -thieno)[2,3-d]pyrimidines 3a-d, 4a-d, 5a-c and 6a-c.

The novel preparation of 2‐cyanobutanamide derivatives starting from 2‐amino‐4,5‐dihydro‐3‐furancarbonitriles is described. Thermal treatment of 4,5‐dihydro‐2‐(4‐morpholinyl, 1‐piperidinyl, and 1‐pyrrolidinyl)‐3‐furancarbonitriles with various carboxylic acids in the absence of solvent caused a ring‐opening reaction to give the corresponding 2‐cyanobutanamide derivatives.

A novel series of 6-substituted 4-sulfonyl-1,4-diazepane-2,5-diones were designed, synthesized and evaluated as human chymase inhibitors. Structure-activity relationship studies led to the identification of a potent inhibitor, (6S)-6-(5-chloro-2-methoxybenzyl)-4-[(4-chlorophenyl)sulfonyl]-1,4-diazepane-2,5-dione, with an IC(50) of 0.027 microM.

An approach to indole derivatives from N-arylhydroxamic acids and malononitrile via a [3,3]-sigmatropic rearrangement and intramolecular cyclization is described. Reactions of N-arylhydroxamic acids 1a-c, 2a-c and 3a-c with malononitrile in the presence of triethylamine at room temperature gave the corresponding α-cyanoacetamide derivatives 4a-c, 5a-c, 6a-c, 7a-c and 8a-c. Thermal treatment of 4a-c, 5a-c and 7a-c with a base, e.g. triethylamine and sodium methoxide, caused intramolecular cyclization and deacylation to afford the corresponding indole derivatives 9-11.

The purpose of this study was to examine the effect on oxidative stress (OS) of muscle stress caused by different training sets. Twenty-five healthy volunteers (22.0 ± 2.6 years old) with no past history of medication were recruited for this study. A free radical analytical system was used to measure OS (hydroperoxide consistency), and blood specimens were collected from volunteers' finger tips on three occasions: 1) at rest, 2) immediately after test completion, and 3) thirty minutes after test completion. Muscle Stress was assessed by an isokinetic testing and training system, and eccentric contraction was carried out in different training sets (10 times × 5 sets, 5R; and 10 times × 10 sets, 10R) for right knee extension. OS showed a significant increase under 10R (p

In order to identify the group of muscles related to deterioration of balance function in the elderly, an exploratory experiment was conducted to analyze and study the influence of lower limb muscular fatigue on balance disturbance in the stable upright position. A stabilometer and a multipurpose muscle function testing and training system were used to assess 12 normal healthy subjects. Evaluations were made using the total trajectory length and the trajectory length per unit area with the eyes-open for 30 seconds. All statistical analyses were performed using SPSS Ver. 13.0 with analysis of variance (repeated measures) at a significance level of below 5%. In the gluteus maximus muscles, the total trajectory length showed a significant downward trend while the trajectory length per unit area showed a significant upward trend.

Structural optimization of 2-aminonicotinamide derivatives as ghrelin receptor inverse agonists is reported. So as to avoid mechanism-based inactivation (MBI) of CYP3A4, 1,3-benzodioxol ring of the lead compound was modified. Improvement of the main activity and lipophilicity was achieved simultaneously, leading to compound 18a, which showed high lipophilic ligand efficiency (LLE) and low MBI activity.

A series of 2-alkylamino nicotinamide analogs was prepared as orally active ghrelin receptor (ghrelinR) inverse agonists. Starting from compound 1, oral bioavailability was improved by modifying metabolically unstable sites and reducing molecular weight. Brain-permeable compound 33 and compound 24 with low brain permeability were tested in rat models of obesity; 30 mg/kg of compound 33 suppressed weight gain. PK/PD analysis revealed that the anti-obesity effect of ghrelinR inverse agonists depends on their brain concentrations.