DAPT (GSI-IX) is a novel γ-secretase inhibitor, which inhibits Aβ production with IC50 of 20 nM in HEK 293 cells.
Upper; Effect of DAPT, a Notch inhibitor on Notch4-ICD expression in TAMR-MCF-7 cells. Lower; Effect of DAPT on cell proliferation of TAMR-MCF-7 cells. Cells were exposed to DAPT (0.3-10 μM) and cell proliferation was measured at different time points by MTT assay. Data represent mean ± SD with 6 different samples.
cancer lett, 2017, 390:115-125. DAPT (GSI-IX) purchased from Selleck.
A panel of GICs was treated with the indicated doses of DAPT for 48 hours. γSecretase inhibitors inhibited expression of NICD, Hes1, Hes3, and Hes5 in a dose-dependent manner.
Stem Cells 2014 32(1), 301-12. DAPT (GSI-IX) purchased from Selleck.
Effects of endosulfan on cytoskeleton and mitosis in HUVECs. Images 7–12 are the 4 × magnified versions of 1–6, respectively. Microfilament (A), microtubule (B), and cell nucleus (C) were incubated with Actin-Tracker Green, Tubulin-Tracker Red, and Hoechst 33258 solution, respectively.
Environ Pollut, 2017, 221:26-36. DAPT (GSI-IX) purchased from Selleck.
R26PR;cre tumors express high levels of NICD and are sensitive to pharmacological inhibition of NOTCH1 signaling. (C) A cell line derived from R26PR;MMTV-cre tumor cells was cultured in the presence of a γ-secretase inhibitor, DAPT, or DMSO vehicle. Live cells were counted at 24, 48 and 72 hours of culture. (D) Western blot analysis of NICD following DAPT treatment.
Dis Model Mech 2013 6(6), 1494-506. DAPT (GSI-IX) purchased from Selleck.
Human corneal epithelial cells were subjected to a scratch assay and then treated with DAPT or DMSO (control) (A). The effect of DAPT concentration on scratch assay wound closure rate was measured (P < 0.001) (B). Western blot for Notch1IC confirmed that 10uM DAPT can effectively inhibit Notch activation (C). HCE-T cells pretreated with DAPT migrated 2.2 times faster than control in transwell migration assay (P < 0.0001) while Jagged1 treated cells migrated 20% slower but did not reach statistical significance (P =0.077) (D).
Invest Ophth Vis Sci 2012 53,12 . DAPT (GSI-IX) purchased from Selleck.
|製品説明||DAPT (GSI-IX) is a novel γ-secretase inhibitor, which inhibits Aβ production with IC50 of 20 nM in HEK 293 cells.|
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.  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. 
|体内試験||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.  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. |
In vitro Aβ reduction assays :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.
|体外||DMSO||86 mg/mL (198.86 mM)|
|Ethanol||50 mg/mL (115.61 mM)|
4% DMSO+corn oil
質量 (g) = 濃度 (mol/L) x 体積 (L) x 分子量 (g/mol)
開始濃度 x 開始体積 = 最終濃度 x 最終体積
この方程式は、一般に略語を使われます：C1V1 = C2V2 ( 入力 出力 )
チップス: 化学式は大文字と小文字の区別ができます。C10H16N2O2 c10h16n2o2
|NCT Number||Recruitment||Conditions||Sponsor/Collaborators||Start Date||Phases|
|NCT03355742||Recruiting||Bleeding Disorder|Stroke Ischemic|Stroke Hemorrhagic|Hematological Disease|Thrombocytopenia|Coagulation Disorder|Anemia|Renal Insufficiency|Coronary Artery Disease||Abbott Vascular||February 9 2018||Not Applicable|
|NCT03606642||Not yet recruiting||Coronary Artery Disease|Atherosclerosis|Stent Placement||HonorHealth Research Institute|Boston Scientific Corporation||July 23 2018||Phase 2|
|NCT03647475||Not yet recruiting||Coronary Artery Disease||Medtronic Vascular||October 2018||Not Applicable|
|NCT03008083||Not yet recruiting||Drug-Eluting Stents|Percutaneous Coronary Intervention||Shanghai MicroPort Medical (Group) Co. Ltd.||August 2018||Phase 4|
|NCT02798913||Unknown status||Peripheral Artery Disease||Federico II University||January 2016||Phase 3|
|NCT02619760||Active not recruiting||Coronary Artery Disease||Kyoto University Graduate School of Medicine||December 2015||Phase 4|
Could you please help test the formulation of S2215 for in vivo studies?
S2215 DAPT in 30% PEG400+0.5% Tween80+5% Propylene glycol at 10 mg/ml is a suspension. We tried to add some EtOH, and it dissolved clearly in organice solvents, but when water added, the precipitation went out immediately. Then we tried other vehicles, and found S2215 can be dissolved in 4% DMSO+corn oil at 10 mg/ml clearly.
I would like to ask if you would recommend this product used in endothelial cells (e.g. both murine and human endothelial cells).
I think DAPT can be used in endothelial cells from both human and mouse, please see the following reference: http://www.ncbi.nlm.nih.gov/pubmed/19481797; http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2615564/