SAR405
mRNA synthesis
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
Inhibitor Cocktails
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
SAR405是一种选择性的、ATP竞争性的Vps34抑制剂,Kd值为1.5 nM。
Vps34是磷酸肌醇3-激酶(PI3K)的第III类亚型,在自噬中有重要作用。Vps34最早在酵母中被发现参与囊泡运输。SAR405是第一个Vps34催化活性的抑制剂,可作为一个独特的药理工具用于研究该蛋白的生物学功能。由于SAR405独特的结合模式以及与人Vps34的ATP结合域的分子间相互作用,SAR405具有蛋白和脂酶的选择性。SAR405通过抑制Vps34激酶活性,从而导致晚期内涵体-溶酶体积累并阻止自噬的发生。但是,SAR405并不会影响早期的内吞作用。同时,SAR405会引起cathepsin D成熟的缺陷,表明溶酶体的功能受损。这与早期的报道相一致:Vps34 siRNA会影响从晚期内涵体到溶酶体的囊泡运输。
SAR405抑制人重组Vps34对PtdIns的磷酸化,IC50值为1 nM。GFP-FYVE的重定位表明SAR405抑制PtdIns3P的形成。SAR405达到10 μM时对I型、II型PI3Ks和mTOR均没有效果。在PC3细胞中,SAR405浓度达到10 μM时并不影响Akt的磷酸化。SAR405(1 μM)可以与来自Jurkat细胞裂解物的Vps34蛋白ATP结合位点以及脂酶相结合,其KD值是重组Vps34的600倍。
参考文献:
[1]. Ronan B, Flamand O1, Vescovi L et al. A highly potent and selective Vps34 inhibitor alters vesicle trafficking and autophagy. Nat Chem Biol. 2014 Dec;10(12):1013-9.
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Storage | Store at -20°C |
M.Wt | 443.85 |
Cas No. | 1523406-39-4 |
Formula | C19H21ClF3N5O2 |
Solubility | ≥22.19 mg/mL in DMSO; insoluble in H2O; ≥32.25 mg/mL in EtOH with ultrasonic |
Chemical Name | (S)-1-((5-chloropyridin-3-yl)methyl)-8-((R)-3-methylmorpholino)-2-(trifluoromethyl)-3,4-dihydro-1H-pyrimido[1,2-a]pyrimidin-6(2H)-one |
SDF | Download SDF |
Canonical SMILES | C[C@H]1N(C(N=C2N3CC[C@@H](C(F)(F)F)N2CC4=CN=CC(Cl)=C4)=CC3=O)CCOC1 |
运输条件 | 蓝冰运输或根据您的需求运输。 |
一般建议 | 不同厂家不同批次产品溶解度各有差异,仅做参考。若实验所需浓度过大至产品溶解极限,请添加助溶剂助溶或自行调整浓度。溶液形式一般不宜长期储存,请尽快用完。 |
激酶实验[1]: | |
Vps34 细胞实验 |
在专用的Vps34细胞实验中评价SAR405的活性。使用GFP-FYVE转染的HeLa细胞系,SAR405的治疗引起GFP-FYVE在细胞内的再定位而不影响GFP强度。1 μM温育后测量SAR405与来自Jurkat细胞裂解物的蛋白质和脂质激酶的ATP位点的结合,其KD为重组Vps34的600倍以上。结果证实SAR405与内源性Vps34有效的结合,产生大于 94%的抑制。 |
细胞实验: [1] | |
细胞系 |
GFP-LCLC3 HeLa 细胞,GFP-LCLC3 H1299细胞 |
制备方法 |
该化合物在DMSO中的溶解度大于10 mM。若配制更高浓度的溶液,一般步骤如下:请将试管置于37℃加热10分钟和/或将其置于超声波浴中震荡一段时间。原液于-20℃可放置数月。 |
反应条件 |
16 h-24 h |
实验结果 |
在RKO细胞中,SAR405处理24小时导致p62蛋白的剂量依赖性积累,SAR405处理16小时成熟组织蛋白酶D显著减少。SAR405也完全抑制GFP-LCLC3 HeLa细胞中自噬体的形成。此外,在GFP-LCLC3 H1299细胞中,SAR405阻止由mTOR抑制剂诱导的自噬,并与Everolimus 具有协同性。 |
References: 1. Ronan B, Flamand O1, Vescovi L et al. A highly potent and selective Vps34 inhibitor alters vesicle trafficking and autophagy. Nat Chem Biol. 2014 Dec;10(12):1013-9. |
质量控制和MSDS
- 批次: