PHT-427

 

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

Biological Activity

Description	PHT-427 is a dual Akt and PDPK1 inhibitor (high affinity binding for the PH domains of Akt and PDPK1) with Kiof 2.7 μM and 5.2 μM, respectively.
Targets	Akt	PDPK1
IC50	2.7 μM	5.2 μM [1]
In vitro	PH-427 is a pleckstrin homology domain inhibitor to Akt/PDPK1. PH-427 significantly reduces phospho-Ser241-PDPK1 phospho-Thr308-Akt in PC-3 prostate cancer cells at 10 μM, which shows that PHT-427 could inhibit both Akt and PDKP1. PHT-427 also inhibits translocation of the Akt and PDKP1 PH domains in plasma membrane. [1] PHT-427 induces apoptosis and inhibits AKT phosphorylation with IC50 of 8.6 μM (in BxPC-3 cells), which mainly on its Ser473 residue and less strongly on Thr308 residue without affecting total Akt protein expression. PHT-427 also shows antiproliferation in Panc-1 cells with IC50 of 65 μM. [2]
In vivo	PHT-427 shows great antitumor activity in BxPC-3 pancreatic, MCF-7 breast and A-549 NSCL cancer xenografts. PHT-427 gives up to an 80% inhibition of tumor growth in BxPC-3 at doses of 125 to 250 mg/kg. [1]
Features

Protocol(Only for Reference)

Kinase Assay: ^[1]

Surface plasmon resonance (SPR) spectroscopy binding assays

All interaction analyses are performed with a Biacore 2000, Biacore 2000 Control Software v3.2, and BIAevaluation v4.1 analysis software. The PH domain GST-fusion proteins (Akt1, IRS1, and PDK1) are immobilized on a CM5 Sensorchip using Biacores Amine Coupling Kit to a level of 10,000 Response units (RUs). Small molecule analytes at concentrations ranging from 0.1 to 10 × the predicted KD are injected at a high flow rate (30μL/min). DMSO concentrations in all samples and running buffer are 1% (v/v) or less.

Cell Assay: ^[2]

Cell lines	Panc-1 cells
Concentrations	1-50 μM
Incubation Time
Method	Cell growth inhibition is determined using a micro-cytoxicity assay. Cells are plated in 96-well micro-cytoxicity at 5-10 × 103 cells per well (depending on cell doubling time) and grown for 7 days. PHT-427 dissolved in DMSO is added directly to the media, at various concentrations ranging from 1 to 50 μM. The endpoint is spectrophotometric determination of the protein content of each well using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. A concentration-response relationship at two or more concentration levels is used to obtain an IC50 for PHT-427.

Animal Study: ^[1]

Animal Models	BxPC-3, Panc-1, MiaPaCa-2, PC-3, SKOV-3, A-549 or MCF-7 cells are injected subcutaneously into the flanks of female scid mice.
Dosages	125-250 mg/kg
Administration	Oral administration
Solubility	5% DMSO/95% Corn oil, 5 mg/mL

1

References

Chemical Information

Download PHT-427 SDF

Chemical Name	4-dodecyl-N-(1,3,4-thiadiazol-2-yl)benzenesulfonamide

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.40961	4.0961	8.1922	12.2883

Customer Reviews (1)

Click to enlarge	Rating
	Source	BLOOD 2011 118, 2840-2848. PHT-427 purchased from Selleck
	Method	Luciferase reporter assays
	Cell Lines	32D cells, BCR-ABL cells
	Concentrations	20 μM
	Incubation Time	24 h
	Results	Treatment of 32D/BCR-ABL cells withthe AKT/PDPK1 inhibitor PHT-427, the PI3K inhibitor LY294002 or the BCR-ABL inhibitor imatinib substantially reduced Atf5 promoter-directed luciferase activity.

Product Citations (1)

• 
  

  BCR-ABL suppresses autophagy through ATF5-mediated regulation of mTOR transcription. [Sheng Z, et al. Blood 2011;118(10):2840-8]

  
  PubMed: 21715304