Home Proteases Secretase inhibitor - Beta Amyloid inhibitor - Autophagy modulator - Apoptosis related activator - Notch inhibitor DAPT For research use only.

DAPT

Synonyms: GSI-IX, LY-374973

DAPT is a novel γ-secretase inhibitor, which inhibits Aβ production with IC50 of 20 nM in HEK 293 cells. DAPT enhances the apoptosis of human tongue carcinoma cells and regulates autophagy.

DAPT Chemical Structure

DAPT Chemical Structure

CAS No. 208255-80-5

Purity & Quality Control

Products often used together with DAPT

GI254023X

DAPT and GI254023X block NOTCH activation in PC3 cells co-cultured with OP9-DLL1/OP9 cells.

Jouannet S, et al. Cell Mol Life Sci. 2016 May;73(9):1895-915.

SU5402

DAPT and SU5402, along with other small molecule inhibitors, accelerate the derivation of functional, early-born cortical neurons from human pluripotent stem cells (hPSCs).

Qi Y, et al. Nature biotechnology 35.2 (2017): 154-163.

SB431542

DAPT and SB431542, along with other small molecules, efficiently reprogram cultured human fetal astrocytes into functional neurons.

Ma NX, et al. Frontiers in Cell and Developmental Biology 7 (2019): 82.

Y-27632 2HCl

DAPT and Y-27632 2HCl, along with other small molecule compounds, are used for In vitro stimulation of muller (MCs)/TR-MUL5 cells.

Fujii Y, et al. PLoS One. 2023 Feb 23;18(2):e0282174.

DAPT Related Products

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
Function assay THP1 Displacement of [3H]IN973 from gamma-secretase in human THP1 cells, acvalue. 17932033
GC-B  Growth Inhibition Assay 6.25-100 μM 24 h DMSO inhibits the cell growth in a dose-dependent manner 19542446
U87  Function Assay 2 μM 48 h DMSO blocks t-AUCB-induced activation of the p38 MAPK/MAPKAPK2/Hsp27 pathway and inhibits expression of NICD1 24793313
A549  Growth Inhibition Assay 10 μM 24h decreases the cell viability combined with PTE 23671619
U87  Growth Inhibition Assay 2 μM 48 h DMSO strengthens t-AUCB-induced cell growth suppression 24793313
U251 Function Assay 2 μM 48 h DMSO blocks t-AUCB-induced activation of the p38 MAPK/MAPKAPK2/Hsp27 pathway and inhibits expression of NICD1 24793313
U251 Growth Inhibition Assay 2 μM 48 h DMSO strengthens t-AUCB-induced cell growth suppression 24793313
Saos-2 Function Assay 100 μM 24 h DMSO desensitizes the cell line to cisplatin treatment 24894297
MG63 Function Assay 100 μM 24 h DMSO desensitizes the cell line to cisplatin treatment 24894297
SHG-44 Growth Inhibition Assay 0.5-10 μM 1-5 d inhibits the cell viability at the optimal concentration of 1 μM 25063285
A549 CD133− Growth Inhibition Assay 2 μM 48 h enhances cell growth inhibition induced by CDDP 24502949
A549 CD133+ Growth Inhibition Assay 2 μM 48 h enhances cell growth inhibition induced by CDDP 24502949
HT29  Growth Inhibition Assay 0.5-75 μM 12/24/48 h DMSO inhibits the cell growth in a concentration manner 25257945
Cytotoxicity assay SNU475 72 hrs Cytotoxicity against human SNU475 cells assessed as growth inhibition after 72 hrs by SRB assay, Displacement of [3H]IN973 from gamma-secretase in human THP1 cells, acvalue.. ChEMBL
Cytotoxicity assay HuH7 72 hrs Cytotoxicity against human HuH7 cells assessed as growth inhibition after 72 hrs by SRB assay, Cytotoxicity against human SNU475 cells assessed as growth inhibition after 72 hrs by SRB assay, Displacement of [3H]IN973 from gamma-secretase in human THP1 cells, acvalue... ChEMBL
Cytotoxicity assay Hep3B 72 hrs Cytotoxicity against human Hep3B cells assessed as growth inhibition after 72 hrs by SRB assay, Cytotoxicity against human HuH7 cells assessed as growth inhibition after 72 hrs by SRB assay, Cytotoxicity against human SNU475 cells assessed as growth inhibition after 72 hrs by SRB assay, Displacement of [3H]IN973 from gamma-secretase in human THP1 cells, acvalue.... ChEMBL
Cytotoxicity assay Mahlavu 72 hrs Cytotoxicity against human Mahlavu cells assessed as growth inhibition after 72 hrs by SRB assay, Cytotoxicity against human Hep3B cells assessed as growth inhibition after 72 hrs by SRB assay, Cytotoxicity against human HuH7 cells assessed as growth inhibition after 72 hrs by SRB assay, Cytotoxicity against human SNU475 cells assessed as growth inhibition after 72 hrs by SRB assay, Displacement of [3H]IN973 from gamma-secretase in human THP1 cells, acvalue..... ChEMBL
Click to View More Cell Line Experimental Data

Biological Activity

Description DAPT is a novel γ-secretase inhibitor, which inhibits Aβ production with IC50 of 20 nM in HEK 293 cells. DAPT enhances the apoptosis of human tongue carcinoma cells and regulates autophagy.
Targets
Notch [1] Aβ [1]
(HEK 293 cells)
20 nM
In vitro
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]
Kinase Assay 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.
Cell Research Cell lines SK-MES-1
Concentrations 2.5 μM to 160 μM
Incubation Time 72 hours
Method 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.
Experimental Result Images Methods Biomarkers Images PMID
Western blot NICD / Pax7 / Pax3 / MyoD / Myogenin / p21 Bax / caspase-3 / Bcl-2 Snail / N-cadherin / Vimentin / E-cadherin 18957511
Growth inhibition assay Cell viability 27118928
Immunofluorescence CDK5 18662245
In Vivo
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]
Animal Research Animal Models Heterozygous PDAPP transgenic mice overexpressing the APPV717F mutant form of the amyloid precursor protein.
Dosages ≤100 mg/kg
Administration Administered via p.o.
NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT06292117 Recruiting
Ischemic Stroke|CYP2C19 Polymorphism
University of Virginia|American Heart Association
 April 16 2024 --
NCT06074549 Recruiting
Coronary Artery Disease
Elixir Medical Corporation
 March 10 2024 --
NCT06223607 Recruiting
Baseline Thrombocytopenia
Methodist Health System
 August 22 2022 --

Chemical Information & Solubility

Molecular Weight 432.46 Formula

C23H26F2N2O4
CAS No. 208255-80-5 SDF Download DAPT SDF
Smiles CC(C(=O)NC(C1=CC=CC=C1)C(=O)OC(C)(C)C)NC(=O)CC2=CC(=CC(=C2)F)F
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 86 mg/mL ( (198.86 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Ethanol : 41 mg/mL

Water : Insoluble


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In vivo
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Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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Frequently Asked Questions

Question 1:
Could you please help test the formulation of S2215 for in vivo studies?

Answer:
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.

Question 2:
I would like to ask if you would recommend this product used in endothelial cells (e.g. both murine and human endothelial cells).

Answer:
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/

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