DAPT (GSI-IX)
(Synonyms: gamma-Secretase Inhibitor IX, DAPT, GSI-IX) 目录号 : GC12942
Inhibitor of γ-secretase
Cas No.:208255-80-5
Sample solution is provided at 25 µL, 10mM.
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| Cell experiment [1]: | |
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Cell lines |
HepG2 cells |
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Preparation Method |
HepG2 cells were treated with different dosages of DAPT (25, 50, and 100 µM, n=3), with HepG2 cells incubated with blank medium as the control group. |
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Reaction Conditions |
25, 50, and 100 µM |
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Applications |
The cell viability of HepG2 cells was significantly inhibited by the introduction of DAPT in a dose-dependent manner. Compared with the control, in the colony formation assay, the colony number was found to be significantly suppressed in the 50 and 100 µM DAPT groups |
| Animal experiment [2]: | |
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Animal models |
Eight-week-old Apoe−/− mice (B6.129P2) |
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Preparation Method |
Mini osmotic pumps containing AngII (1 μg/min/kg) were implanted subcutaneously. At day 14, mice were randomly divided into two groups. A group of mice was administered DAPT (10 mg/kg dissolved in 10% ethanol, 90% corn oil) three times a week for the next 28 days. |
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Dosage form |
10 mg/kg, p.o. |
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Applications |
This 10 mg/kg concentration was effective in reducing the aneurysm formation in AngII-induced mouse model of AAA. |
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References: [1]. Qiu K, et al. DAPT suppresses proliferation and migration of hepatocellular carcinoma by regulating the extracellular matrix and inhibiting the Hes1/PTEN/AKT/mTOR signaling pathway. J Gastrointest Oncol. 2021 Jun;12(3):1101-1116. [2]. Hans CP, et al. DAPT, a potent Notch inhibitor regresses actively growing abdominal aortic aneurysm via divergent pathways. Clin Sci (Lond). 2020 Jun 26;134(12):1555-1572. |
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DAPT (GSI-IX) is an orally active γ-secretase inhibitor with IC50s of 115 nM and 200 nM for total amyloid-β (Aβ) and Aβ42 produced in human primary neuronal cultures, respectively.[1]
In vitro experiment it indicated that treatment with 10 μM DAPT significantly reduced the expression of Il6, Il12 and iNos at 6 and 12 h compared with vehicle at both low- and high-LPS stimulation.[2] In vitro, with 25, 50, and 100 μM DAPT in HepG2 cells significantly inhibited the proliferation and migration ability of HepG2 cells.[3] In vitro test shown it that treatment with 0, 25, 50 and 75 μM DAPT in CNE-2 cells, pre-treatment with DAPT enhanced the effect of cisplatin in a dose-dependent manner. However, the CNE-2 cells were treated with increasing concentrations of DAPT, and there was no obvious effect on cell survival.[5]
In vivo efficacy study it shown that treatment with 100 mg/kg DAPT subcutaneously in PDAPP mice, after 3 h the peak level of DAPT were 490 ng/g in the brain, and levels greater than 100 ng/g were sustained throughout the first 18 h.[1] DAPT (10, 30 and 100 mg/kg; p.o.) reduced the cortical total Aβ in a dose-dependent manner with a 50% reduction occuring at 100 mg/kg dosing.[1] In vivo test indicated it that DAPT was administered intragastrically once daily for 28 days in ICR mice that can effectively in ameliorating Cd-induced multi-organ damage and cognitive impairment in mice, because of DAPT restored abnormal performance in the Y-maze, forced swimming and Morris water maze (MWM) tests.[4]
References:
[1].Dovey HF, et al. Functional gamma-secretase inhibitors reduce beta-amyloid peptide levels in brain. J Neurochem. 2001 Jan;76(1):173-81.
[2].Hans CP, et al. DAPT, a potent Notch inhibitor regresses actively growing abdominal aortic aneurysm via divergent pathways. Clin Sci (Lond). 2020 Jun 26;134(12):1555-1572.
[3].Qiu K, et al. DAPT suppresses proliferation and migration of hepatocellular carcinoma by regulating the extracellular matrix and inhibiting the Hes1/PTEN/AKT/mTOR signaling pathway. J Gastrointest Oncol. 2021 Jun;12(3):1101-1116.
[4].Yang JY, et al. DAPT Attenuates Cadmium-Induced Toxicity in Mice by Inhibiting Inflammation and the Notch/HES-1 Signaling Axis. Front Pharmacol. 2022 Apr 29;13:902796.
[5].Zhou JX, et al. γ-secretase inhibition combined with cisplatin enhances apoptosis of nasopharyngeal carcinoma cells. Exp Ther Med. 2012 Feb;3(2):357-361.
DAPT (GSI-IX) 是一种具有口服活性的 γ-分泌酶抑制剂，对人原代神经元培养物中产生的总淀粉样蛋白-β (Aβ) 和 Aβ42 的 IC50 分别为 115 nM 和 200 nM。[1]
体外实验表明，在低和高 LPS 刺激下，与载体相比，10 μM DAPT 在 6 和 12 h 显着降低了 Il6、Il12 和 iNos 的表达。[2] 在体外，25、50、100 μM DAPT对HepG2细胞有显着抑制HepG2细胞增殖和迁移能力的作用。[3] 体外试验表明，0、25、 50 和 75 μM DAPT 在 CNE-2 细胞中，用 DAPT 预处理以剂量依赖的方式增强了顺铂的作用。然而，CNE-2细胞经增加浓度的DAPT后，对细胞存活无明显影响。[5]
体内功效研究表明，在 PDAPP 小鼠中皮下注射 100 mg/kg DAPT，3 小时后大脑中 DAPT 的峰值水平为 490 ng/g，并且在整个过程中持续保持大于 100 ng/g 的水平前 18 小时。[1] DAPT（10、30 和 100 mg/kg；口服）以剂量依赖性方式降低皮质总 Aβ，在 100 mg/kg 时降低 50% [1] 体内试验表明，ICR小鼠每天1次灌胃DAPT，连续给药28天，可有效改善Cd诱导的小鼠多器官损伤和认知障碍，因为DAPT 恢复了 Y 迷宫、强迫游泳和莫里斯水迷宫 (MWM) 测试中的异常表现。[4]
| Cas No. | 208255-80-5 | SDF | |
| 别名 | gamma-Secretase Inhibitor IX, DAPT, GSI-IX | ||
| 化学名 | tert-butyl (2S)-2-[[(2S)-2-[[2-(3,5-difluorophenyl)acetyl]amino]propanoyl]amino]-2-phenylacetate | ||
| Canonical SMILES | CC(C(=O)NC(C1=CC=CC=C1)C(=O)OC(C)(C)C)NC(=O)CC2=CC(=CC(=C2)F)F | ||
| 分子式 | C23H26F2N2O4 | 分子量 | 432.46 |
| 溶解度 | ≥ 21.623mg/mL in DMSO, ≥ 16.36 mg/mL in EtOH with ultrasonic | 储存条件 | Store at 4°C |
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.3124 mL | 11.5618 mL | 23.1235 mL |
| 5 mM | 0.4625 mL | 2.3124 mL | 4.6247 mL |
| 10 mM | 0.2312 mL | 1.1562 mL | 2.3124 mL |
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HIF-1α is Critical for the Activation of Notch Signaling in Neurogenesis During Acute Epilepsy
Emerging evidence suggests that hypoxia-inducible factors (specifically, HIF-1α) and Notch signaling are involved in epileptogenesis and that cross-coupling exists between HIF-1α and Notch signaling in other diseases, including tumors and ischemia. However, the exact molecular mechanisms by which HIF-1α and Notch signaling affect the development of epilepsy, especially regarding neurogenesis, remain unclear. In the present study, we investigated the role of HIF-1α in neurogenesis and whether Notch signaling is involved in this process during epileptogenesis by assessing hippocampal apoptosis, neuronal injury, and the proliferation and differentiation of neural stem cells (NSCs) in four groups, including control, epilepsy, epilepsy+2-methoxyestradiol (2ME2) and epilepsy+GSI-IX (DAPT) groups. Our data demonstrated that HIF-1α mediated neurogenesis during acute epilepsy, which required the participation of Notch signaling. The immunoprecipitation data illustrated that HIF-1α activated Notch signaling by physically interacting with the Notch intracellular domain (NICD) in epilepsy. In conclusion, our results suggested that HIF-1α-Notch signaling enhanced neurogenesis in acute epilepsy and that neurogenesis during epileptogenesis was reduced once this pathway was blocked; thus, members of this pathway might be potential therapeutic targets for epilepsy.
Notch signaling pathway is a potential therapeutic target for extracranial vascular malformations
Notch expression has been shown to be aberrant in brain arteriovenous malformations (AVM), and targeting Notch has been suggested as an approach to their treatment. It is unclear whether extracranial vascular malformations follow the same patterning and Notch pathway defects. In this study, we examined human extracranial venous (VM) (n = 3), lymphatic (LM) (n = 10), and AV (n = 6) malformations, as well as sporadic brain AVMs (n = 3). In addition to showing that extracranial AVMs demonstrate interrupted elastin and that AVMs and LMs demonstrate abnormal α-smooth muscle actin just as brain AVMS do, our results demonstrate that NOTCH1, 2, 3 and 4 proteins are overexpressed to varying degrees in both the endothelial and mural lining of the malformed vessels in all types of malformations. We further show that two gamma secretase inhibitors (GSIs), DAPT (GSI-IX) and RO4929097, cause dose-dependent inhibition of Notch target gene expression (Hey1) and rate of migration of monolayer cultures of lymphatic endothelial cells (hLECs) and blood endothelial cells (HUVEC). GSIs also inhibit HUVEC network formation. hLECs are more sensitive to GSIs compared to HUVEC. GSIs have been found to be safe in clinical trials in patients with Alzheimer's disease or cancer. Our results provide further rationale to support testing of Notch inhibitors in patients with extracranial vascular malformations.
γ-Secretase inhibitor reduces immunosuppressive cells and enhances tumour immunity in head and neck squamous cell carcinoma
Immune evasion is a hallmark feature of cancer, and it plays an important role in tumour initiation and progression. In addition, tumour immune evasion severely hampers the desired antitumour effect in multiple cancers. In this study, we aimed to investigate the role of the Notch pathway in immune evasion in the head and neck squamous cell carcinoma (HNSCC) microenvironment. We first demonstrated that Notch1 signaling was activated in a Tgfbr1/Pten-knockout HNSCC mouse model. Notch signaling inhibition using a γ-secretase inhibitor (GSI-IX, DAPT) decreased tumour burden in the mouse model after prophylactic treatment. In addition, flow cytometry analysis indicated that Notch signaling inhibition reduced the sub-population of myeloid-derived suppressor cells (MDSCs), tumour-associated macrophages (TAMs) and regulatory T cells (Tregs), as well as immune checkpoint molecules (PD1, CTLA4, TIM3 and LAG3), in the circulation and in the tumour. Immunohistochemistry (IHC) of human HNSCC tissues demonstrated that elevation of the Notch1 downstream target HES1 was correlated with MDSC, TAM and Treg markers and with immune checkpoint molecules. These results suggest that modulating the Notch signaling pathway may decrease MDSCs, TAMs, Tregs and immune checkpoint molecules in HNSCC.
NOTCH1 inhibition enhances the efficacy of conventional chemotherapeutic agents by targeting head neck cancer stem cell
Cancer stem cells (CSCs) are considered responsible for tumor initiation and chemoresistance. This study was aimed to investigate the possibility of targeting head neck squamous cell carcinoma (HNSCC) by NOTCH1 pathway inhibition and explore the synergistic effect of combining NOTCH inhibition with conventional chemotherapy. NOTCH1/HES1 elevation was found in human HNSCC, especially in tissue post chemotherapy and lymph node metastasis, which is correlated with CSCs markers. NOTCH1 inhibitor DAPT (GSI-IX) significantly reduces CSCs population and tumor self-renewal ability in vitro and in vivo. Flow cytometry analysis showed that NOTCH1 inhibition reduces CSCs frequency either alone or in combination with chemotherapeutic agents, namely, cisplatin, docetaxel, and 5-fluorouracil. The combined strategy of NOTCH1 blockade and chemotherapy synergistically attenuated chemotherapy-enriched CSC population, promising a potential therapeutic exploitation in future clinical trial.
Blocking the NOTCH pathway can inhibit the growth of CD133-positive A549 cells and sensitize to chemotherapy
Cancer stem cells (CSCs) are believed to play an important role in tumor growth and recurrence. These cells exhibit self-renewal and proliferation properties. CSCs also exhibit significant drug resistance compared with normal tumor cells. Finding new treatments that target CSCs could significantly enhance the effect of chemotherapy and improve patient survival. Notch signaling is known to regulate the development of the lungs by controlling the cell-fate determination of normal stem cells. In this study, we isolated CSCs from the human lung adenocarcinoma cell line A549. CD133 was used as a stem cell marker for fluorescence-activated cell sorting (FACS). We compared the expression of Notch signaling in both CD133+ and CD133- cells and blocked Notch signaling using the γ-secretase inhibitor DAPT (GSI-IX). The effect of combining GSI and cisplatin (CDDP) was also examined in these two types of cells. We observed that both CD133+ and CD133- cells proliferated at similar rates, but the cells exhibited distinctive differences in cell cycle progression. Few CD133+ cells were observed in the G2/M phase, and there were half as many cells in S phase compared with the CD133- cells. Furthermore, CD133+ cells exhibited significant resistance to chemotherapy when treated with CDDP. The expression of Notch signaling pathway members, such as Notch1, Notch2 and Hes1, was lower in CD133+ cells. GSI slightly inhibited the proliferation of both cell types and exhibited little effect on the cell cycle. The inhibitory effects of DPP on these two types of cells were enhanced when combined with GSI. Interestingly, this effect was especially significant in CD133+ cells, suggesting that Notch pathway blockade may be a useful CSC-targeted therapy in lung cancer.