DAPT

Biological Activity

DAPT (GSI-IX) is a γ-secretase inhibitor with IC50 values of 115 and 200 nM for total Aβ and Aβ42 respectively. DAPT (GSI-IX) may be useful in the study of β-amyloid (Aβ) formation. As an inhibitor of Notch, DAPT may advance studies of autoimmune and lymphoproliferative diseases, such as ALPS and lupus erythematosus (SLE). Other γ-secretase substrates include LDL receptor-related protein, E-cadherin and ErbB-4.

Product Citations

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  Glycoconj J . 2022 Jun 6.

  Notch/NICD/RBP-J signaling axis regulates M1 polarization of macrophages mediated by advanced glycation end products
  DAPT purchased from AbMole

• 

  2015 May.

  Acquisition of resistance to trastuzumab in gastric cancer cells is associated with activation of IL-6/STAT3/Jagged-1/Notch positive feedback loop
  DAPT purchased from AbMole

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  J Ethnopharmacol. 2019 Oct 28;243:112092.

  Study of the aqueous extract of Aloe vera and its two active components on the Wnt/β-catenin and Notch signaling pathways in colorectal cancer cells.
  DAPT purchased from AbMole

• 

  Oncol Lett. 2017 July 5;4: 3616-3622.

  The role of Jagged1/Notch pathway‑mediated angiogenesis of hepatocarcinoma cells in vitro, and the effects of the spleen‑invigorating and blood stasis‑removing recipe
  DAPT purchased from AbMole

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  Circ Res. 2017 Jul 31.

  SUMOylation Negatively Regulates Angiogenesis by Targeting Endothelial NOTCH Signaling
  DAPT purchased from AbMole

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  Sci Adv. 2015 Apr 10.

  PlGF-induced VEGFR1-dependent vascular remodeling determines opposing antitumor effects and drug resistance to Dll4-Notch inhibitors.
  DAPT purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Oncol Lett (2017). Figure 6. DAPT (AbMole BioScience, Shanghai, China)
	Method	Co‑IP assay
	Cell Lines	SMMC‑7721 cells
	Concentrations	1 μM
	Incubation Time
	Results	The interaction between Jagged1 and Notch1 in SMMC‑7721 cells was also significantly reduced after adding DAPT compared with that of the normal serum group (P<0.01) (Fig. 6).

	Source	Oncol Lett (2017). Figure 5. DAPT (AbMole BioScience, Shanghai, China)
	Method	Western blot
	Cell Lines	SMMC‑7721 cells
	Concentrations	1 μM
	Incubation Time
	Results	The levels of Jagged1 and Notch1 were decreased after adding the Notch inhibitor, DAPT, indicating that DAPT successfully inhibited the Jagged1/Notch signaling pathway. In addition, the expression of VEGF protein was reduced after adding DAPT (Fig. 5).

	Source	Circ Res (2017). Figure 2. DATP (Abmole Bioscience)
	Method	Retina dissection and whole mount staining
	Cell Lines	mice
	Concentrations	100 mg/kg
	Incubation Time
	Results	DAPT treatment resulted in a large increase in vessel density and expansion of the vascular plexus in the postnatal retina of control mice (Fig. 2A, C, E, G). DAPT completely rescued the defect in SENP1-ecKO pups (Fig. 2B, D, F, H), which resulted in vascular length, vascular coverage, and tip sprouts growth comparable to the control group (Fig. 2I-K).

	Source	Sci adv (2015). Figure 5. The DAPT g-secretase inhibitor was obtained from AbMole BioScience.
	Method	Immunohistochemistry
	Cell Lines
	Concentrations	100 mg/kg, daily
	Incubation Time
	Results	"The PlGF-modulated opposing vascular effects in relation to Dll4-Notch inhibition alter tumor growth by changing the microenvironment, tumor cell proliferation, and apoptosis."

	Source	Sci adv (2015). Figure 4. The DAPT g-secretase inhibitor was obtained from AbMole BioScience.
	Method	human tumor models, Immunohistochemistry
	Cell Lines
	Concentrations	100 mg/kg, daily
	Incubation Time
	Results	PlGF mediates opposing vascular effects in nontreated and Dll4-Notch inhibitor–treated mouse tumors.

	Source	Sci adv (2015). Figure 3. The DAPT g-secretase inhibitor was obtained from AbMole BioScience.
	Method	Immunohistochemistry
	Cell Lines	Human JE-3, BeWo, MDA-MB-231 cells
	Concentrations	100 mg/kg, daily
	Incubation Time
	Results	"Both terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end labeling (TUNEL) staining and measurements of the activated or cleaved caspase 3 showed reductions of apoptotic cells in Dll4-Notch inhibitor–treated JE-3 and BeWo tumors. In contrast, treatment of PlGF-negative MDAMB-231 tumors with DAPT and Dll4 blockade showed completely opposite effects on cell proliferation and apoptosis, tipping the balance toward an apoptotic phenotype."

	Source	Sci adv (2015). Figure 1. The DAPT g-secretase inhibitor was obtained from AbMole BioScience.
	Method	Immunohistochemistry
	Cell Lines	Human JE-3, BeWo, MDA-MB-231 cells
	Concentrations	100 mg/kg, daily
	Incubation Time
	Results	"PlGF expression determines opposing effects of Dll4-Notch inhibition on human tumor growth and vascular remodeling."

Protocol (for reference only)

Cell Experiment
Cell lines	SK-MES-1
Preparation 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.
Concentrations	2.5 ~ 160 μM
Incubation time	72 h

Animal Experiment
Animal models	three‐ to four‐month‐old heterozygous PDAPP transgenic mice overexpressing the APPV717F mutant form of the amyloid precursor protein
Formulation	formulated in corn oil, 5% (v/v) ethanol
Dosages	100 mg/kg
Administration	s.c.

Chemical Information

Molecular Weight	432.46
Formula	C23H26F2N2O4
CAS Number	208255-80-5
Solubility (25°C)	DMSO 76 mg/mL
Storage	Powder          -20°C   3 years ;  4°C   2 years In solvent       -80°C   6 months ;  -20°C   1 month

Conversion of different model animals based on BSA (PMID: 27057123)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

References

[1] Kim BK et al. J Am Coll Cardiol. A New Strategy for Discontinuation of Dual Antiplatelet Therapy: The RESET Trial (REal Safety and Efficacy of 3-month dual antiplatelet Therapy following Endeavor zotarolimus-eluting stent implantation).

[2] Kedhi E et al. EuroIntervention. Stent thrombosis: insights on outcomes, predictors and impact of dual antiplatelet therapy interruption from the SPIRIT II, SPIRIT III, SPIRIT IV and COMPARE trials.

[3] Oikawa et al. Neurosci Lett. The γ-secretase inhibitor DAPT increases the levels of gangliosides at neuritic terminals of differentiating PC12 cells.

[4] Zou et al. Clin Exp Med. Gamma-secretase inhibitor DAPT suppresses glioblastoma growth via uncoupling of tumor vessel density from vessel function.

[5] H F Dovey, et al. J Neurochem. Functional gamma-secretase inhibitors reduce beta-amyloid peptide levels in brain