Summary of Study ST001135
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000760. The data can be accessed directly via it's Project DOI: 10.21228/M8FH6C This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST001135 |
| Study Title | Different dose exposure of OPC-163493 on HepG2 cells (part-I) |
| Study Type | Compound dosage test |
| Study Summary | Metabolomics analysis were on 8 samples of HepG2 cells that were treated with compound OPC-163493 (DMSO control, 1, 3, or 10µM; each n=2) exposure for 30 min. |
| Institute | Otsuka Pharmaceutical Co., Ltd. |
| Last Name | Kanemoto |
| First Name | Naohide |
| Address | 463-10 Kagasuno Kawauchi-cho Tokushima 771-0192, Japan |
| Kanemoto.Naohide@otsuka.jp | |
| Phone | 81-03-6717-1400 |
| Submit Date | 2019-02-07 |
| Analysis Type Detail | LC-MS |
| Release Date | 2019-03-06 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR000760 |
| Project DOI: | doi: 10.21228/M8FH6C |
| Project Title: | Antidiabetic and cardiovascular beneficial effects of a liver-localized mitochondrial uncoupler |
| Project Summary: | Inducing mitochondrial uncoupling (mUncoupling) is an attractive therapeutic strategy for treating metabolic diseases because it leads to calorie-wasting by reducing the efficiency of oxidative phosphorylation (OXPHOS) in mitochondria. Here we report a safe mUncoupler, OPC-163493, which has unique pharmacokinetic characteristics. OPC-163493 shows a good bioavailability upon oral administration and primarily distributed to specific organs: the liver and kidneys, avoiding systemic toxicities. It exhibitsinsulin-independent antidiabetic effects in multiple animal models of type I and type II diabetes and antisteatotic effects in fatty liver models. These beneficial effects can be explained by the improvement of glucose metabolism and enhancement of energy expenditure by OPC-163493 in the liver. Moreover, OPC-163493 treatment lowered blood pressure, extended survival, and improved renal function in the rat model of stroke/hypertension, possibly by enhancing NO bioavailability in blood vessels and reducing mitochondrial ROS production. OPC-163493 is a liver-localized/targeted mUncoupler that ameliorates various complications of diabetes. |
| Institute: | Otsuka Pharmaceutical Co., Ltd. |
| Last Name: | Kanemoto |
| First Name: | Naohide |
| Address: | 463-10 Kagasuno Kawauchi-cho, Tokushima, Tokusima, 770-0865, Japan |
| Email: | Kanemoto.Naohide@otsuka.jp |
| Phone: | +81-88-665-2126 |
Subject:
| Subject ID: | SU001196 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Not applicable |
| Cell Strain Details: | HepG2 cells |
Factors:
Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | Treatment |
|---|---|---|
| SA078229 | OPC-0uM-5 | - |
| SA078230 | OPC-0uM-1 | - |
| SA078231 | OPC-1uM-2 | 1 |
| SA078232 | OPC-1uM-6 | 1 |
| SA078235 | OPC-10uM-8 | 10 |
| SA078236 | OPC-10uM-4 | 10 |
| SA078233 | OPC-3uM-3 | 3 |
| SA078234 | OPC-3uM-7 | 3 |
| Showing results 1 to 8 of 8 |
Collection:
| Collection ID: | CO001190 |
| Collection Summary: | Culture medium of HepG2 cells in 100-mm dish was aspirated and cells were washed twice by 5% mannitol solution, and then processed with sampleprep steps. |
| Sample Type: | HepG2 cells |
| Storage Conditions: | Room temperature |
Treatment:
| Treatment ID: | TR001211 |
| Treatment Summary: | HepG2 cells were seeded into a 100mm dish the day before OPC-163493 treatment. OPC-163493 treatments (DMSO control, 1, 3, or 10uM, each n=2) were performed for 30min in FBS-free DMEM with high glucose (25mM). |
| Treatment Dose: | 0uM, 1uM, 3uM, 10uM |
| Treatment Doseduration: | 30 min |
| Cell Media: | MEM with Earle`s salts, L-Glutamine and Non-Essiontial Amino Acids, 10% fetal bovine serum, and 1mM sodium pyruvate solution |
Sample Preparation:
| Sampleprep ID: | SP001204 |
| Sampleprep Summary: | Culture medium of HepG2 cells in 100-mm dish was aspirated and cells were washed twice by 5% mannitol solution, and then the cells were treated with methanol. The cell extract was treated with Mili-Q water containing internal standard and filtered with 5-kDa cutoff filter. The filtrate was centrifugally concentrated and re-suspended in 50 µL of Milli-Q water. |
| Processing Storage Conditions: | Room temperature |
| Extract Storage: | -80℃ |
| Sample Resuspension: | 50 uL Mili-Q |
Combined analysis:
| Analysis ID | AN001860 | AN001861 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | CE | CE |
| Chromatography system | Agilent 7100 CE | Agilent 7100 CE |
| Column | Fused silica capillary, i.d. 50 μm × 80 cm | Fused silica capillary, i.d. 50 μm × 80 cm |
| MS Type | ESI | ESI |
| MS instrument type | TOF | Triple quadrupole |
| MS instrument name | Agilent 6210 TOF | Agilent 6460 QQQ |
| Ion Mode | UNSPECIFIED | UNSPECIFIED |
| Units | Concentration (pmol/1000000 cells) | Concentration (pmol/1000000 cells) |
Chromatography:
| Chromatography ID: | CH001347 |
| Chromatography Summary: | capillary electrophoresis was connected with time-of-flight mass spectrometry (CE-TOFMS) for cation analysis and tandem mass spectrometry (CE-MS/MS) for anion. |
| Instrument Name: | Agilent 7100 CE |
| Column Name: | Fused silica capillary, i.d. 50 μm × 80 cm |
| Chromatography Type: | CE |
MS:
| MS ID: | MS001720 |
| Analysis ID: | AN001860 |
| Instrument Name: | Agilent 6210 TOF |
| Instrument Type: | TOF |
| MS Type: | ESI |
| MS Comments: | The spectrometer was scanned from m/z 50 to 1,000. Peaks were extracted using automatic integration software MasterHands (Keio University, Tsuruoka, Japan) in order to obtain peak information including m/z, migration time for CE-TOFMS measurement (MT) and peak area. Signal peaks corresponding to isotopomers, adduct ions, and other product ions of known metabolites were excluded, and remaining peaks were annotated with putative metabolites from the HMT metabolite database based on their MTs and m/z values determined by TOFMS. The tolerance range for the peak annotation was configured at ±0.5 min for MT and ±10 ppm for m/z. In addition, peak areas were normalized against those of the internal standards and then the resultant relative area values were further normalized by sample amount. |
| Ion Mode: | UNSPECIFIED |
| MS ID: | MS001721 |
| Analysis ID: | AN001861 |
| Instrument Name: | Agilent 6460 QQQ |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | The spectrometer was scanned from m/z 50 to 1,000. Peaks were extracted using MasterHands, automatic integration software (Keio University, Tsuruoka, Yamagata, Japan) and MassHunter Quantitative Analysis B.04.00 (Agilent Technologies) in order to obtain peak information including m/z, peak area, and migration time (MT). Signal peaks were annotated according to the HMT metabolite database based on their m/z values with the MTs. The peak area of each metabolite was normalized with respect to the area of the internal standard and metabolite concentration was evaluated by standard curves with three-point calibrations using each standard compound. |
| Ion Mode: | UNSPECIFIED |
