DAPT (GSI-IX)

Catalog No.S2215 Synonyms: LY-374973

For research use only.

DAPT (GSI-IX, LY-374973) 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 (GSI-IX) Chemical Structure

CAS No. 208255-80-5

Selleck's DAPT (GSI-IX) has been cited by 301 publications

Purity & Quality Control

Choose Selective Secretase Inhibitors

Biological Activity

Description DAPT (GSI-IX, LY-374973) 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
Secretase [1]
(HEK 293 cells)
20 nM
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]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
Function assay NIrQc45VUFBz MoPySIl{eGyjY3Xt[Y51KG:oIGuzTH1KVjl5MzDmdo9uKGejbX3hMZNm[3KndHHz[UBqdiCqdX3hckBVUFBzIHPlcIx{NCCjY4\hcJVmNg>? MlLCQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTd7M{KwN|MoRjF5OUOyNFM{RC:jPh?=
GC-B  M3TEcGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MoH0Ok4zPS1zMECg{txO NYXmZ4F6OjRiaB?= MYrEUXNQ NXPjenNWcW6qaXLpeJMhfGinIHPlcIwh\3Kxd4ToJIlvKGFiZH;z[U1l\XCnbnTlcpQhdWGwbnXy M{XONVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF7NUSyOFQ3Lz5zOUW0NlQ1PjxxYU6=
U87  Mm\GSpVv[3Srb36gRZN{[Xl? M3XEVVIh|ryP MoHoOFghcA>? M3excmROW09? Mnn4Zoxw[2u|wrD0MWFWS0JvaX7keYNm\CCjY4TpeoF1cW:wIH;mJJRp\SCyM{igUWFRUy:PQWDLRXBMOi:Kc4CyO{Bx[XSqd3H5JIFv\CCrbnjpZol1eyCneIDy[ZN{cW:wIH;mJG5KS0Rz NUXvZlN{RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS3PVM{OTNpPkK0O|k{OzF|PD;hQi=>
A549  MmP6S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? MnPZNVAh|ryP MVeyOIg> MkK3[IVkemWjc3XzJJRp\SClZXzsJJZq[WKrbHn0fUBkd22kaX7l[EB4cXSqIGDUSS=> MWW8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOzZ5MU[xPUc,OjN4N{G2NVk9N2F-
U87  NX;ydpJ3T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MX2yJO69VQ>? NFfZUWk1QCCq MXjEUXNQ NYHYcpQze3S{ZX7neIhmdnQEoIStRXVESi2rbnT1Z4VlKGOnbHyg[5Jwf3SqIIP1dJBz\XO|aX;u MUS8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDd7M{OxN{c,OjR5OUOzNVM9N2F-
U251 NH\JNJRHfW6ldHnvckBCe3OjeR?= NU[z[GZXOiEQvF2= M1nLelQ5KGh? NYrNUIg1TE2VTx?= Mof5Zoxw[2u|wrD0MWFWS0JvaX7keYNm\CCjY4TpeoF1cW:wIH;mJJRp\SCyM{igUWFRUy:PQWDLRXBMOi:Kc4CyO{Bx[XSqd3H5JIFv\CCrbnjpZol1eyCneIDy[ZN{cW:wIH;mJG5KS0Rz M3m0TlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2N{mzN|E{Lz5{NEe5N|MyOzxxYU6=
U251 NIOwWY1Iem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NIPLNWMzKM7:TR?= M1HvNVQ5KGh? MV\EUXNQ MlPWd5Rz\W6pdHjlcpPDqHRvQWXDRk1qdmS3Y3XkJINmdGxiZ4Lve5RpKHO3cIDy[ZN{cW:w NHvwS2k9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NEe5N|MyOyd-MkS3PVM{OTN:L3G+
Saos-2 M2rjTGZ2dmO2aX;uJGF{e2G7 M4DBU|ExOCEQvF2= M3riN|I1KGh? M3fNPWROW09? NH\6VJZl\XOnboPpeIl7\XNidHjlJINmdGxibHnu[UB1dyClaYPwcIF1cW5idILlZZRu\W62 MXq8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDh7NEK5O{c,OjR6OUSyPVc9N2F-
MG63 MoXmSpVv[3Srb36gRZN{[Xl? MYqxNFAh|ryP M1yzS|I1KGh? NVLHW4YzTE2VTx?= MVXk[ZNmdnOrdHn6[ZMhfGinIHPlcIwhdGmwZTD0c{BkcXOybHH0bY4hfHKnYYTt[Y51 NGrG[Zo9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NEi5OFI6Pyd-MkS4PVQzQTd:L3G+
SHG-44 NVi1N41lT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NILrXZQxNjVvMUCg{txO Mn3oNU02KGR? M2j2PIlvcGmkaYTzJJRp\SClZXzsJJZq[WKrbHn0fUBifCC2aHWgc5B1cW2jbDDjc45k\W62cnH0bY9vKG:oIEGg{txO Ml;KQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjVyNkOyPFUoRjJ3ME[zNlg2RC:jPh?=
A549 CD133− MnXlS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NFjSNFQzKM7:TR?= NFfifo01QCCq M2Hxc4VvcGGwY3XzJINmdGxiZ4Lve5RpKGmwaHnibZRqd25iaX7keYNm\CCkeTDDSGRR MlXnQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjR3MEK5OFkoRjJ2NUCyPVQ6RC:jPh?=
A549 CD133+ NV3qW3R6T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= M2TmO|Ih|ryP MYW0PEBp Mnjr[Y5p[W6lZYOgZ4VtdCCpcn;3eIghcW6qaXLpeIlwdiCrbnT1Z4VlKGK7IFPESHA> M3PQNVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2NUCyPVQ6Lz5{NEWwNlk1QTxxYU6=
HT29  M3rXZmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NEjqNpAxNjVvN{Wg{txO NI\ZW|IyOi9{ND:0PEBp MYLEUXNQ NUSxNIdGcW6qaXLpeJMhfGinIHPlcIwh\3Kxd4ToJIlvKGFiY3;uZ4VvfHKjdHnvckBu[W6wZYK= MYG8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPTJ3N{m0OUc,OjV{NUe5OFU9N2F-
Cytotoxicity assay M3TKT3NPXTR5NR?= M1nhVVczKGi{cx?= MWjDfZRwfG:6aXPpeJkh[WejaX7zeEBpfW2jbjDTUnU1PzViY3XscJMh[XO|ZYPz[YQh[XNiZ4Lve5RpKGmwaHnibZRqd25iYX\0[ZIhPzJiaILzJIJ6KFOUQjDhd5NigSxiRHnzdIxi[2WvZX70JI9nKFt|SG3JUlk4OyCocn;tJIdidW2jLYPlZ5JmfGG|ZTDpckBpfW2jbjDUTHAyKGOnbHzzMEBi[3[jbIXlMk4> MYS8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyyOVU3QDJxJ{7DbGVOSkx:L3G+
Cytotoxicity assay Mn\VTJVJPw>? NELtfYM4OiCqcoO= M3rCO2N6fG:2b4jpZ4l1gSCjZ3HpcpN1KGi3bXHuJGh2UDdiY3XscJMh[XO|ZYPz[YQh[XNiZ4Lve5RpKGmwaHnibZRqd25iYX\0[ZIhPzJiaILzJIJ6KFOUQjDhd5NigSxiQ4n0c5RwgGmlaYT5JIFo[Wmwc4SgbJVu[W5iU17VOFc2KGOnbHzzJIF{e2W|c3XkJIF{KGe{b4f0bEBqdmirYnn0bY9vKGGodHXyJFczKGi{czDifUBUWkJiYYPzZZktKESrc4DsZYNmdWWwdDDv[kBcO0ifSV65O|Mh\nKxbTDnZY1u[S2|ZXPy[ZRie2ViaX6gbJVu[W5iVFjQNUBk\WyuczygZYN3[Wy3ZT6uMi=> NXrnXZBmRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNOjV3NkiyM{c,S2iHTVLMQE9iRg>?
Cytotoxicity assay M3q5TGhmeDOE MmrzO|IhcHK| MlTqR5l1d3SxeHnjbZR6KGGpYXnud5QhcHWvYX6gTIVxO0JiY3XscJMh[XO|ZYPz[YQh[XNiZ4Lve5RpKGmwaHnibZRqd25iYX\0[ZIhPzJiaILzJIJ6KFOUQjDhd5NigSxiQ4n0c5RwgGmlaYT5JIFo[Wmwc4SgbJVu[W5iSIXIO{Bk\WyuczDhd5Nme3OnZDDhd{Boem:5dHigbY5pcWKrdHnvckBi\nSncjC3NkBpenNiYomgV3JDKGG|c3H5MEBEgXSxdH;4bYNqfHliYXfhbY5{fCCqdX3hckBUVlV2N{WgZ4VtdHNiYYPz[ZN{\WRiYYOg[5Jwf3SqIHnubIljcXSrb36gZYZ1\XJiN{KgbJJ{KGK7IGPSRkBie3OjeTygSIl{eGyjY3Xt[Y51KG:oIGuzTH1KVjl5MzDmdo9uKGejbX3hMZNm[3KndHHz[UBqdiCqdX3hckBVUFBzIHPlcIx{NCCjY4\hcJVmNi5wLh?= NYfsemM{RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNOjV3NkiyM{c,S2iHTVLMQE9iRg>?
Cytotoxicity assay NWO5UW94VWGqbHH2eS=> M1i2WFczKGi{cx?= MnO3R5l1d3SxeHnjbZR6KGGpYXnud5QhcHWvYX6gUYFpdGG4dTDj[YxteyCjc4Pld5Nm\CCjczDndo94fGhiaX7obYJqfGmxbjDh[pRmeiB5MjDodpMh[nliU2LCJIF{e2G7LDDDfZRwfG:6aXPpeJkh[WejaX7zeEBpfW2jbjDI[ZA{SiClZXzsd{Bie3Onc4Pl[EBieyCpcn;3eIghcW6qaXLpeIlwdiCjZoTldkA4OiCqcoOgZpkhW1KEIHHzd4F6NCCFeYTveI95cWOrdImgZYdicW6|dDDoeY1idiCKdVi3JINmdGy|IHHzd4V{e2WmIHHzJIdzd3e2aDDpcohq[mm2aX;uJIFnfGW{IEeyJIhzeyCkeTDTVmIh[XO|YYmsJGN6fG:2b4jpZ4l1gSCjZ3HpcpN1KGi3bXHuJHNPXTR5NTDj[YxteyCjc4Pld5Nm\CCjczDndo94fGhiaX7obYJqfGmxbjDh[pRmeiB5MjDodpMh[nliU2LCJIF{e2G7LDDEbZNxdGGlZX3lcpQhd2ZiW{PIYWlPQTd|IH\yc40h\2GvbXGtd4VkemW2YYPlJIlvKGi3bXHuJHRJWDFiY3XscJMtKGGldnHseYUvNi5wLh?= MVK8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyyOVU3QDJxJ{7DbGVOSkx:L3G+
Assay
Methods Test Index PMID
Western blot Snail / N-cadherin / Vimentin / E-cadherin ; Bax / caspase-3 / Bcl-2 ; NICD / Pax7 / Pax3 / MyoD / Myogenin / p21 24932308 29487808 18957511
Growth inhibition assay Cell viability 27118928
Immunofluorescence CDK5 18662245
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]

Protocol (from reference)

Kinase Assay:[1]
  • 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:[2]
  • 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.
Animal Research:[1]
  • 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.

Solubility (25°C)

In vitro

In vivo

Add solvents to the product individually and in order
(Data is from Selleck tests instead of citations):
4% DMSO+corn oil
For best results, use promptly after mixing.

10mg/mL

Chemical Information

Molecular Weight 432.46
Formula

C23H26F2N2O4

CAS No. 208255-80-5
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles CC(C(=O)NC(C1=CC=CC=C1)C(=O)OC(C)(C)C)NC(=O)CC2=CC(=CC(=C2)F)F

In vivo Formulation Calculator (Clear solution)

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Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

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

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Molarity Calculator

Mass Concentration Volume Molecular Weight

Clinical Trial Information

NCT Number Recruitment Interventions Conditions Sponsor/Collaborators Start Date Phases
NCT04842838 Not yet recruiting Device: paclitaxel DCB|Device: DES Coronary Artery Disease Peking University Third Hospital June 30 2021 Not Applicable
NCT04470934 Recruiting Device: SeQuent® SCB drug-coated balloon catheter Coronary Artery Disease|Myocardial Ischaemia B. Braun Melsungen AG April 30 2021 --
NCT04549766 Not yet recruiting Other: calculate PRECISE DAPT score using clinical data ST-segment Elevation Myocardial Infarction (STEMI) Assiut University September 2020 --
NCT04475536 Recruiting Device: TANSEI stent Ischemic Heart Disease|Coronary Artery Disease Fundación EPIC August 21 2020 --

(data from https://clinicaltrials.gov, updated on 2022-01-17)

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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