Home Proteases Secretase inhibitor DAPT

연구용

DAPT γ-Secretase inhibitor

제품 번호: S2215

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 Secretase inhibitor Chemical Structure

화학 구조

분자량: 432.46

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품질 관리 (Quality Control)

배치: 순도: 99.99%
99.99

함께 자주 사용되는 제품 DAPT

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세포 배양, 처리 및 작업 농도
(Cell Culture, Treatment & Working Concentration)

세포주 분석 유형 농도 배양 시간 제형 활성 설명 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
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화학 정보, 보관 및 안정성 (Chemical Information, Storage & Stability)

분자량 432.46 화학식

C23H26F2N2O4

 보관 (수령일로부터)
CAS 번호 208255-80-5 SDF 다운로드 원액 보관

동의어 GSI-IX, LY-374973 Smiles CC(C(=O)NC(C1=CC=CC=C1)C(=O)OC(C)(C)C)NC(=O)CC2=CC(=CC(=C2)F)F

용해도 (Solubility)

In vitro
배치:

DMSO : 86 mg/mL (198.86 mM)
(수분으로 오염된 DMSO는 용해도를 감소시킬 수 있습니다. 신선하고 무수 DMSO를 사용하십시오.)

Ethanol : 41 mg/mL

Water : Insoluble

몰농도 계산기

질량 농도 부피 분자량
희석 계산기 분자량 계산기

In vivo
배치:

생체 내 제형 계산기 (투명한 용액)

1단계: 아래 정보 입력 (권장: 실험 중 손실을 고려하여 추가 동물 포함)

mg/kg g μL

2단계: 생체 내 제형 입력 (이것은 계산기일 뿐 제형이 아닙니다. 용해도 섹션에 생체 내 제형이 없는 경우 먼저 당사에 문의하십시오.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

계산 결과:

작업 농도: mg/ml;

DMSO 원액 준비 방법: mg 약물 사전 용해 μL DMSO ( 원액 농도 mg/mL, 농도가 해당 약물 배치의 DMSO 용해도를 초과하는 경우 먼저 당사에 문의하십시오. )

생체 내 제형 준비 방법: 취하다 μL DMSO 원액, 다음 추가μL PEG300, 혼합하고 투명하게 한 다음 추가μL Tween 80, 혼합하고 투명하게 한 다음 추가 μL ddH2O, 혼합하고 투명하게 합니다.

생체 내 제형 준비 방법: 취하다 μL DMSO 원액, 다음 추가 μL 옥수수 기름, 혼합하고 투명하게 합니다.

참고: 1. 다음 용매를 추가하기 전에 액체가 투명한지 확인하십시오.
2. 용매를 순서대로 추가해야 합니다. 다음 용매를 추가하기 전에 이전 추가에서 얻은 용액이 투명한 용액인지 확인해야 합니다. 와동, 초음파 또는 뜨거운 물 중탕과 같은 물리적 방법을 사용하여 용해를 도울 수 있습니다.

작용 메커니즘 (Mechanism of Action)

Targets/IC50/Ki
Notch
Aβ
(HEK 293 cells)
20 nM
시험관 내(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. A recent study shows that this compound inhibits the proliferation of SK-MES-1 cells in a concentration-dependent manner with IC50 of 11.3 μM. In addition, it also induces caspase-dependent and caspase-independent apoptosis in lung squamous cell carcinoma cells by inhibiting Notch receptor signaling pathway.
키나아제 분석
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 this compound, 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 this chemical. Cultures are then treated with fresh media containing this compound 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 this compound concentration, data are analyzed with XLfit software, as above, to determine potency.
생체 내(In vivo)
DAPT administration (100mg/kg) leads to a robust and sustained pharmacodynamic effect in PDAPP mice that this compound 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, this chemical (100 mg/kg) also reduces the cortical total Aβ and Aβ42 in a dose-dependent manner with a 50% reduction. In rat cerebral cortexes, this compound (40 mg/kg) suppresses the LPS-induced activity of γ-secretase and increases the cell apoptosis with the prolonged neuroinflammation.
참조

적용 분야 (Applications)

방법 바이오마커 이미지 PMID
Western blot NICD / Pax7 / Pax3 / MyoD / Myogenin / p21 Bax / caspase-3 / Bcl-2 Snail / N-cadherin / Vimentin / E-cadherin
S2215-WB3
18957511
Growth inhibition assay Cell viability
S2215-viability1
27118928
Immunofluorescence CDK5
S2215-IF1
18662245

임상시험 정보 (Clinical Trial Information)

(데이터 출처 https://clinicaltrials.gov, 업데이트 날짜 2024-05-22)

NCT 번호 모집 조건 스폰서/협력자 시작일 단계
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 --

자주 묻는 질문 (Frequently Asked Questions)

질문 1:
I would like to ask if you would recommend it used in endothelial cells (e.g. both murine and human endothelial cells).

답변:
I think this compound 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/