Molecule

Description

Cancer

Description

TGX-221 is a potent, selective PI3K inhibitor for p110β with IC50 of 8.5 nM.

Targets

IC₅₀

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 ± 13.9%) and 3+3 (88 ± 10.6%) 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]

Clinical Trials

Features

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 Lines

PC3 cells

Concentrations

~20 μM

Incubation Time

24-72 hours

Methods

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 Models

FeCl3-induced arterial thrombosis in mice

Formulation

TGX-221 is dissolved in 10% ethanol, 10% cremaphor, 10% N,N-dimethylacetamine, 70% distilled water.

Doses

0.3 + 0.3, 1 + 1, 3 + 3 mg/kg + mg/kg/hour

Administration

Administered via i.v.