化合物信息

Description

Cancer

Description

PHT-427 is a dual Akt and PDPK1 inhibitor with K_i of 2.7 μM and 5.2 μM, respectively.

Targets

IC₅₀

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 6.3 μM, 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]

Clinical Trials

Features

Description

Combined with paclitaxel (10 mg/kg), PHT-427 (200 mg/kg) exhibits greater antitumor activity than both monotherapy in MCF-7 xenograft. Combined with erlotinib (50 mg/kg), PHT-427 (200 mg/kg) significantly inhibits tumor growth in NCI-H441 xenograft, which is negative to erlotinib. ^[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 Biacore’s 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 Lines

Panc-1 cells

Concentrations

1-50 μM

Incubation Time

Methods

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 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.

Formulation

40 to 50 mg/mL in sesame seed oil

Doses

125-250 mg/kg

Administration

Oral administration