TGX-221

 Distributor Tel: +886-2-2827-1197jimmy@new.abscience.com.tw

Biological Activity

Description	TGX-221 is a p110β-specific inhibitor with IC50 of 5 nM, 1000-fold more selective for p110β than p110α.
Targets	p110β
IC50	5 nM [1]
In vitro	The activity of TGX-221 against different isoforms is measured in an in vitro PI3K assay using multiple preparations of recombinant p85/p110. TGX-221 show slow potent to p110δ with IC50 of 211 nM. Furthermore, TGX-221 partially attenuates insulin-induced phosphorylation of Ser473 of PKB in J774.2 macrophage cells. [1]TGX-221 inhibits platelet-ECC interaction, platelet aggregation and platelet-granulocyte binding in an extracorporeal circulation (ECC) model. [2] A recent study shows that after treatment with TGX-221 (0.2, 2, and 20 μM), PC3 cells show inhibition of proliferation with a significant reduction of the activity of the p110β PI3K isoform. [3]
In vivo	As an anti-thrombotic agent, TGX-221 at doses 1 + 1 (49 %) and 3+3 (88 %) improves integrated blood flow over 30 minutes in a mouse model. In addition, Tail bleeding time (BT) (sec) increases with TGX-221 doses of 3 + 3 (median 1560) and 1 + 1 (1305) and mean renal BT (sec) also increases in all TGX-221 groups. [4]
Features

Protocol(Only for Reference)

Kinase Assay: ^[1]

Lipid kinase activity

IC50 values are measured using a standard lipid kinase activity with PI as a substrate. (i)100 μM cold ATP is used instead of 10 μM, (ii) the DMSO concentration is 1%, and (iii) [γ-33P]ATP is used instead of [γ-32P]ATP. The TLC plates are quantified using a phosphorimager screen. The reported IC50 values are determined by non-linear regression analysis on the basis of at least three independent experiments repeated across multiple preparations of recombinant protein.

Cell Assay: ^[3]

Cell lines	PC3 cells
Concentrations	~20 μM
Incubation Time	24-72 hours
Method	For measurement of proliferation, cells are seeded in triplicate in 96-well culture plates and incubated overnight to allow cell attachment. The cells are incubated with TGX-221 for 24, 48, and 72 hours. At designated time intervals, cells are quantified by a crystal violet staining-based colorimetric assay. Briefly, cells are fixed by addition of 100 μl of 2.5% glutaraldehyde solution and incubated at room temperature for 30 minutes. Plates are washed three times by submersion in PBS solution. Plates are air-dried and stained by addition of 100 μL of 0.1% solution of crystal violet dissolved in deionized water and incubated for 20 minutes at room temperature, excess dye is removed by extensive washing with deionized water, and plates are air-dried prior to bound dye solubilization in 100 μL of 10% acetic acid. The optical density of dye extracts is measured directly in plates using a microplate reader at 570 nm.

Animal Study: ^[3]

Animal Models	FeCl3-induced arterial thrombosis in mice
Dosages	0.3 + 0.3, 1 + 1, 3 + 3 mg/kg + mg/kg/hour
Administration	Administered via i.v.
Solubility	1% DMSO/30% polyethylene glycol/1% Tween 80, 30 mg/mL

1

References

Chemical Information

Download TGX-221 SDF

Chemical Name	7-methyl-2-morpholino-9-(1-(phenylamino)ethyl)-4H-pyrido[1,2-a]pyrimidin-4-one

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.36444	3.6444	7.2888	10.9332

Customer Reviews (5)

Click to enlarge	Rating
	Source	Mol Med, 2012, 18, 336-45. TGX-221 purchased from Selleck
	Method	Western blot/ELISA PI3K Assays
	Cell Lines	synaptoneurosomes
	Concentrations	1 μmol/L
	Incubation Time	30 min
	Results	TGX-221 reduces p110β-specific PI3K activity and Akt phosphorylation in both WT and Fmr1 KO SNS(Figure A). Quantification of an ELISA-based p110β -specific PI3K activity assay in WT and Fmr1KO SNS after TGX-221 treat-ment shows a significant decrease in both genotypes (Figure B). downstream signaling is similarly reduced after TGX-221 treatment, as shown by densitometric quan-tification of phosphoAkt-specific Western blots (Figure C)

Product Citations (5)

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  Phosphoinositide 3 kinase inhibition restores neutrophil accuracy in the elderly: towards targeted treatments for immunesenescence. [Sapey E, et al. Blood 2014;123(2):239-49]

  
  PubMed: 24191150


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  Excess protein synthesis in FXS patient lymphoblastoid cells can be rescued with a p110β-selective inhibitor. [Gross C, et al. Mol Med 2012;18:336-45]

  
  PubMed: 22207187


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  Interferon-γ activates transglutaminase 2 via a phosphatidylinositol-3-kinase-dependent pathway: implications for celiac sprue therapy. [Diraimondo TR, et al. J Pharmacol Exp Ther 2012;341(1):104-14]

  
  PubMed: 22228808