Name: DAPT (GSI-IX)
Brand: Selleck Chemicals
SKU: S2215
Rating: 5 (293 reviews)

CAS No. 208255-80-5

Selleck's DAPT (GSI-IX) has been cited by 293 publications

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Gamma-secretase Signaling Pathway Map

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

γ secretase(Aβ) ^[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	NFK5NJBVUFBz				MoO5SIl{eGyjY3Xt[Y51KG:oIGuzTH1KVjl5MzDmdo9uKGejbX3hMZNm[3KndHHz[UBqdiCqdX3hckBVUFBzIHPlcIx{NCCjY4\hcJVmNg>?	MlzSQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTd7M{KwN\|MoRjF5OUOyNFM{RC:jPh?=
GC-B	Mmj4S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?	M4nFeFYvOjVvMUCwJO69VQ>?	Mmn6NlQhcA>?	NFPLW4tFVVOR	MlzmbY5pcWKrdIOgeIhmKGOnbHyg[5Jwf3SqIHnuJIEh\G:\|ZT3k[ZBmdmSnboSgcYFvdmW{	NGHMdZQ9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zOUW0NlQ1Pid-MUm1OFI1PDZ:L3G+
U87	NGf1To1HfW6ldHnvckBCe3OjeR?=	MV[yJO69VQ>?	Mn\pOFghcA>?	MWLEUXNQ	Mm\MZoxw[2u\|wrD0MWFWS0JvaX7keYNm\CCjY4TpeoF1cW:wIH;mJJRp\SCyM{igUWFRUy:PQWDLRXBMOi:Kc4CyO{Bx[XSqd3H5JIFv\CCrbnjpZol1eyCneIDy[ZN{cW:wIH;mJG5KS0Rz	NFToZZI9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NEe5N\|MyOyd-MkS3PVM{OTN:L3G+
A549	NXPDWpFrT3Kxd4ToJGlvcGmkaYTpc44hSXO\|YYm=	NYL5eoQ6OTBizszN	NHPDe\|MzPGh?		M3rUOYRm[3KnYYPld{B1cGViY3XscEB3cWGkaXzpeJkh[2:vYnnu[YQhf2m2aDDQWGU>	NHXKZmE9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{M{[3NVYyQSd-MkO2O\|E3OTl:L3G+
U87	MnPMS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?	NHnTUYIzKM7:TR?=	Mn7vOFghcA>?	M1vz[mROW09?	NH;0cmR{fHKnbnf0bIVve8LidD3BWWNDNWmwZIXj[YQh[2WubDDndo94fGhic4XwdJJme3Orb36=	Mk\xQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjR5OUOzNVMoRjJ2N{mzN\|E{RC:jPh?=
U251	MnPVSpVv[3Srb36gRZN{[Xl?	NHfEVnkzKM7:TR?=	M3\keVQ5KGh?	NGjp[oVFVVOR	NXj2NnN4[myxY3vzxsB1NUGXQ1KtbY5lfWOnZDDhZ5RqfmG2aX;uJI9nKHSqZTDwN\|ghVUGSSz;NRXBMSVCNMj;Id5AzPyCyYYToe4F6KGGwZDDpcohq[mm2czDlfJBz\XO\|aX;uJI9nKE6LQ1Sx	M3ftOFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2N{mzN\|E{Lz5{NEe5N\|MyOzxxYU6=
U251	MVLHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?	M1fGcFIh\|ryP	NFXP[JI1QCCq	NYL6OmhmTE2VTx?=	NWLU[JV2e3S{ZX7neIhmdnQEoIStRXVESi2rbnT1Z4VlKGOnbHyg[5Jwf3SqIIP1dJBz\XO\|aX;u	Ml7BQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjR5OUOzNVMoRjJ2N{mzN\|E{RC:jPh?=
Saos-2	NXrVT5FoTnWwY4Tpc44hSXO\|YYm=	NW\Q[nd4OTByIN88US=>	NWfoOVBbOjRiaB?=	M37tVWROW09?	MnnX[IV{\W6\|aYTpfoV{KHSqZTDj[YxtKGyrbnWgeI8h[2m\|cHzheIlvKHS{ZXH0cYVvfA>?	MnO1QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjR6OUSyPVcoRjJ2OEm0Nlk4RC:jPh?=
MG63	MV\GeY5kfGmxbjDBd5NigQ>?	NIfIVZIyODBizszN	MXmyOEBp	NGDleHRFVVOR	MWnk[ZNmdnOrdHn6[ZMhfGinIHPlcIwhdGmwZTD0c{BkcXOybHH0bY4hfHKnYYTt[Y51	M160ZlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2OEm0Nlk4Lz5{NEi5OFI6PzxxYU6=
SHG-44	NUfuZYd7T3Kxd4ToJGlvcGmkaYTpc44hSXO\|YYm=	Ml7SNE42NTFyIN88US=>	M2HQ[FEuPSCm		NX;MZmFCcW6qaXLpeJMhfGinIHPlcIwhfmmjYnnsbZR6KGG2IITo[UBweHSrbXHsJINwdmOnboTyZZRqd25ib3[gNUDPxE1?	MXy8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPTB4M{K4OUc,OjVyNkOyPFU9N2F-
A549 CD133−	M{f4W2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7	MVGyJO69VQ>?	NGezSHk1QCCq		MWjlcohidmOnczDj[YxtKGe{b4f0bEBqdmirYnn0bY9vKGmwZIXj[YQh[nliQ1TEVC=>	M{XKTVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2NUCyPVQ6Lz5{NEWwNlk1QTxxYU6=
A549 CD133+	NYPlS49tT3Kxd4ToJGlvcGmkaYTpc44hSXO\|YYm=	NYXlNYVUOiEQvF2=	MXm0PEBp		NFryc3VmdmijbnPld{Bk\WyuIHfyc5d1cCCrbnjpZol1cW:wIHnu[JVk\WRiYomgR2RFWA>?	MYS8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDVyMkm0PUc,OjR3MEK5OFk9N2F-
HT29	M4m3Rmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7	NVKx[GgyOC53LUe1JO69VQ>?	Mor3NVIwOjRxNEigbC=>	NUL2cJpkTE2VTx?=	Mo\1bY5pcWKrdIOgeIhmKGOnbHyg[5Jwf3SqIHnuJIEh[2:wY3XueJJifGmxbjDtZY5v\XJ?	MmC0QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjV{NUe5OFUoRjJ3MkW3PVQ2RC:jPh?=
Cytotoxicity assay	NFTV[YJUVlV2N{W=		NIHURpA4OiCqcoO=		NVzDdGk1S3m2b4TvfIlkcXS7IHHnZYlve3RiaIXtZY4hW06XNEe1JINmdGy\|IHHzd4V{e2WmIHHzJIdzd3e2aDDpcohq[mm2aX;uJIFnfGW{IEeyJIhzeyCkeTDTVmIh[XO\|YYmsJGRqe3CuYXPlcYVvfCCxZjDbN2heUU57N{Og[pJwdSCpYX3tZU1{\WO{ZYThd4UhcW5iaIXtZY4hXEiSMTDj[YxteyxiYXP2ZYx2\S5w	NYi3PZhFRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNOjV3NkiyM{c,S2iHTVLMQE9iRg>?
Cytotoxicity assay	M1TIV2h2UDd?		NV3WOoZVPzJiaILz		M1PqdGN6fG:2b4jpZ4l1gSCjZ3HpcpN1KGi3bXHuJGh2UDdiY3XscJMh[XO\|ZYPz[YQh[XNiZ4Lve5RpKGmwaHnibZRqd25iYX\0[ZIhPzJiaILzJIJ6KFOUQjDhd5NigSxiQ4n0c5RwgGmlaYT5JIFo[Wmwc4SgbJVu[W5iU17VOFc2KGOnbHzzJIF{e2W\|c3XkJIF{KGe{b4f0bEBqdmirYnn0bY9vKGGodHXyJFczKGi{czDifUBUWkJiYYPzZZktKESrc4DsZYNmdWWwdDDv[kBcO0ifSV65O\|Mh\nKxbTDnZY1u[S2\|ZXPy[ZRie2ViaX6gbJVu[W5iVFjQNUBk\WyuczygZYN3[Wy3ZT6uMi=>	NGPMdJA9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmwzPTV4OEKvK\|5EcEWPQly8M4E,
Cytotoxicity assay	NHvVU2JJ\XB\|Qh?=		MmToO\|IhcHK\|		NFLteW1EgXSxdH;4bYNqfHliYXfhbY5{fCCqdX3hckBJ\XB\|QjDj[YxteyCjc4Pld5Nm\CCjczDndo94fGhiaX7obYJqfGmxbjDh[pRmeiB5MjDodpMh[nliU2LCJIF{e2G7LDDDfZRwfG:6aXPpeJkh[WejaX7zeEBpfW2jbjDIeWg4KGOnbHzzJIF{e2W\|c3XkJIF{KGe{b4f0bEBqdmirYnn0bY9vKGGodHXyJFczKGi{czDifUBUWkJiYYPzZZktKEO7dH;0c5hq[2m2eTDh[4FqdnO2IHj1cYFvKFOQVUS3OUBk\WyuczDhd5Nme3OnZDDhd{Boem:5dHigbY5pcWKrdHnvckBi\nSncjC3NkBpenNiYomgV3JDKGG\|c3H5MEBFcXOybHHj[Y1mdnRib3[gX\|NJZUmQOUezJIZzd21iZ3HtcYEue2WlcnX0ZZNmKGmwIHj1cYFvKFSKUEGgZ4VtdHNuIHHjeoFtfWVwLj6u	MXG8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyyOVU3QDJxJ{7DbGVOSkx:L3G+
Cytotoxicity assay	NVzlUoY3VWGqbHH2eS=>		NEXEOpk4OiCqcoO=		NFXuNWFEgXSxdH;4bYNqfHliYXfhbY5{fCCqdX3hckBO[WiuYY\1JINmdGy\|IHHzd4V{e2WmIHHzJIdzd3e2aDDpcohq[mm2aX;uJIFnfGW{IEeyJIhzeyCkeTDTVmIh[XO\|YYmsJGN6fG:2b4jpZ4l1gSCjZ3HpcpN1KGi3bXHuJGhmeDOEIHPlcIx{KGG\|c3Xzd4VlKGG\|IHfyc5d1cCCrbnjpZol1cW:wIHHmeIVzKDd{IHjyd{BjgSCVUlKgZZN{[XluIFP5eI91d3irY3n0fUBi\2GrboP0JIh2dWGwIFj1TFch[2WubIOgZZN{\XO\|ZXSgZZMh\3Kxd4ToJIlvcGmkaYTpc44h[W[2ZYKgO\|IhcHK\|IHL5JHNTSiCjc4PhfUwhS3m2b4TvfIlkcXS7IHHnZYlve3RiaIXtZY4hW06XNEe1JINmdGy\|IHHzd4V{e2WmIHHzJIdzd3e2aDDpcohq[mm2aX;uJIFnfGW{IEeyJIhzeyCkeTDTVmIh[XO\|YYmsJGRqe3CuYXPlcYVvfCCxZjDbN2heUU57N{Og[pJwdSCpYX3tZU1{\WO{ZYThd4UhcW5iaIXtZY4hXEiSMTDj[YxteyxiYXP2ZYx2\S5wLj6u	M{XseFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFI2PTZ6Mj:nQmNpTU2ETEyvZV4>

Click to View More Cell Line Experimental Data

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. (Only for Reference)
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. (Only for Reference)

References

Solubility (25°C)

In vitro	DMSO	86 mg/mL (198.86 mM)
	Ethanol	50 mg/mL (115.61 mM)
	Water	Insoluble
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. Click to purchase: Corn Oil	10mg/mL

* 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.

Chemical Information

Molecular Weight	432.46
Formula	C₂₃H₂₆F₂N₂O₄
CAS No.	208255-80-5
Storage	3 years	-20°C	powder
Storage	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

Download DAPT (GSI-IX) SDF

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

Dosage: mg/kg Average weight of animals: g Dosing volume per animal:μL Number of animals:

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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.
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Molarity Calculator

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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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