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化学情報
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Synonyms | GSI-IX, LY-374973 | Storage (From the date of receipt) |
3 years -20°C powder 1 years -80°C in solvent |
| 化学式 | C23H26F2N2O4 |
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| 分子量 | 432.46 | CAS No. | 208255-80-5 | |
| Solubility (25°C)* | 体外 | DMSO | 86 mg/mL (198.86 mM) | |
| Ethanol | 41 mg/mL (94.8 mM) | |||
| Water | Insoluble | |||
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* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. |
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溶剤液（一定の濃度）を調合する
生物活性
| 製品説明 | DAPT (GSI-IX、LY-374973) は新規の γ-セクレターゼ 阻害剤であり、HEK 293 細胞で 20 nM の IC50 で Aβ 産生を阻害します。 DAPT はヒト舌癌細胞のアポトーシス(apoptosis) を促進し、オートファジー (autophagy) を調節します。 |
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| in vitro | In human primary neuronal cultures, DAPT also shows inhibitory effects on Aβ production with IC50 of 115 nM and 200 nM respectively for Aβ total and Aβ42, which is 5-10-fold lower than is observed in HEK 293 cells. [1] A recent study shows that DAPT inhibits the proliferation of SK-MES-1 cells in a concentration-dependent manner with IC50 of 11.3 μM. In addition, DAPT also induces caspase-dependent and caspase-independent apoptosis in lung squamous cell carcinoma cells by inhibiting Notch receptor signaling pathway. [2] |
| in vivo | DAPT administration (100mg/kg) leads to a robust and sustained pharmacodynamic effect in PDAPP mice that DAPT levels in the brain exceeds 100 ng/g within 1 hour and persists up to 18 hours after administration, with peak levels of 490 ng/g observed after 3 hour. And during the period, DAPT (100 mg/kg) also reduces the cortical total Aβ and Aβ42 in a dose-dependent manner with a 50% reduction. [1] In rat cerebral cortexes, DAPT (40 mg/kg) suppresses the LPS-induced activity of γ-secretase and increases the cell apoptosis with the prolonged neuroinflammation. [3] |
プロトコル(参考用のみ)
| キナーゼアッセイ | In vitro Aβ reduction assays | |
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| Human embryonic kidney cells (American Type Culture Collection CRL-1573), transfected with the gene for APP751 (HEK 293) are used for routine Aβ reduction assays. Cells are plated in 96-well plates and allowed to adhere overnight in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum. DAPT are diluted from stock solutions in dimethylsulfoxide (DMSO) to yield a final concentration equal to 0.1% DMSO in media. Cells are pre-treated for 2 hours at 37 °C with DAPT, media are aspirated off and fresh compound solutions applied. After an additional 2-hour treatment period, conditioned media is drawn off and analyzed by a sandwich ELISA (266–3D6) specific for total Aβ. Reduction of Aβ production is measured relative to control cells treated with 0.1% DMSO and expressed as a percentage inhibition. Data from at least six doses in duplicate are fitted to a four-parameter logistical model using XLfit software in order to determine potency. Human and PDAPP mouse neuronal cultures are grown in serum-free media to enhance their neuronal characteristics, and appeared to be greater than 90% neurons after maturation prior to use. Conditioned media to establish baseline Aβ values are collected by adding fresh media to each well and incubated for 24 hours at 37 °C in the absence of DAPT. Cultures are then treated with fresh media containing DAPT at the desired range of concentrations for an additional 24 hours at 37 °C, and conditioned media collected. For the measurement of total Aβ, samples are analyzed with the same ELISA (266–3D6) as used for the HEK 293 cell assays. Analyses of samples for Aβ42 production are performed by a separate ELISA (21F12–3D6) that utilizes a capture antibody specific for the Aβ42 C-terminus. Inhibition of production for both total Aβ and Aβ42 are determined by the difference between the values for the compound treatment and baseline periods. After plotting percentage inhibition versus DAPT concentration, data are analyzed with XLfit software, as above, to determine potency. | ||
| 細胞アッセイ | 細胞株 | SK-MES-1 |
| 濃度 | 2.5 μM to 160 μM | |
| 反応時間 | 72 hours | |
| 実験の流れ | Cells are seeded into 96-well plates and exposed to 0.1% DMSO or DAPT at concentrations in the range of 2.5 μM–160 μM for 72 hours. Cytotoxicity is determined with 3-(4, 5)-dimethylthiahiazo-(-z-y1)-3, 5-di-phenytetrazoliumromide (MTT) dye reduction assay with minor modifications. Briefly, after incubation with DAPT, 20 μL MTT solution (5 mg/mL in PBS) is added to 180 μL medium in each well and plates are incubated for 4 hours at 37 °C, and subsequently 150 μL DMSO is added to each well, and mixed by shaking at room temperature for 15 minutes. Absorption is measured by an enzyme-linked immunosorbent assay at 490 nm to determine absorbance values. α-MEM supplemented with the same amount of MTT solution and solvent is used as blank solution. The IC50 value is calculated using PROBIT program in SPSS. | |
| 動物実験 | 動物モデル | Heterozygous PDAPP transgenic mice overexpressing the APPV717F mutant form of the amyloid precursor protein. |
| 投薬量 | ≤100 mg/kg | |
| 投与方法 | Administered via p.o. | |
参考
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カスタマーフィードバック
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Data from [Data independently produced by Oncogene, 2014, 10.1038/onc.2014.319]
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Data from [Data independently produced by Stem Cells, 2014, 32(1), 301-12]
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Data from [Data independently produced by Dis Model Mech, 2013, 6(6), 1494-506]
Selleckの高級品が、幾つかの出版された研究調査結果（以下を含む）で使われた：
| Mapping multimodal phenotypes to perturbations in cells and tissue with CRISPRmap [ Nat Biotechnol, 2024, 10.1038/s41587-024-02386-x] | PubMed: 39375448 |
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| Pericyte phenotype switching alleviates immunosuppression and sensitizes vascularized tumors to immunotherapy in preclinical models [ J Clin Invest, 2024, 134(18)e179860] | PubMed: 39286984 |
| Mechanosensitive protein polycystin-1 promotes periosteal stem/progenitor cells osteochondral differentiation in fracture healing [ Theranostics, 2024, 14(6):2544-2559] | PubMed: 38646641 |
| HiHo-AID2: boosting homozygous knock-in efficiency enables robust generation of human auxin-inducible degron cells [ Genome Biol, 2024, 25(1):58] | PubMed: 38409044 |
| High glutamine increases stroke risk by inducing the endothelial-to-mesenchymal transition in moyamoya disease [ MedComm (2020), 2024, 5(5):e525] | PubMed: 38628905 |
| Immune evasion in lung metastasis of leiomyosarcoma: upregulation of EPCAM inhibits CD8+ T cell infiltration [ Br J Cancer, 2024, 10.1038/s41416-024-02576-z] | PubMed: 38291183 |
| A splice site variant in MADD affects hormone expression in pancreatic β cells and pituitary gonadotropes [ JCI Insight, 2024, 9(10)e167598] | PubMed: 38775154 |
| Transplanted deep-layer cortical neuroblasts integrate into host neural circuits and alleviate motor defects in hypoxic-ischemic encephalopathy injured mice [ Stem Cell Res Ther, 2024, 15(1):422] | PubMed: 39533375 |
| JAG1 mediates apoptosis in herpes simplex keratitis by suppressing autophagy via ROS/JAG1/NOTCH1/pULK1 signaling pathway [ Cell Biol Toxicol, 2024, 41(1):1] | PubMed: 39704867 |
長期の保管のために-20°Cの下で製品を保ってください。
人間や獣医の診断であるか治療的な使用のためにでない。
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