Description

Cancer

Description

LY2228820 is a novel and potent inhibitor of p38 MAPK with IC50 of 7 nM.

Targets

IC₅₀

In Vitro

LY2228820 inhibits p38α, as well as the level of phosphoMAPKAPK-2 (pMK2) in RAW 264.7 cells, with IC50 values of 7 nM and 34.3 nM, respectively. Furthermore, LY2228820 inhibits lipopolysaccharide (LPS)-induced TNFα formation in murine peritoneal macrophages, with IC50 of 5.2 nM. ^[1]In multiple myeloma (MM) cells, including INA6, RPMI-8226, U266, and RPMI-Dox40, LY2228820 (200 nM–800 nM) significantly blocks p38MAPK signaling, as revealed by its inhibition on phosphorylation of HSP27, a downstream target of p38MAPK, without affecting the expression level of HSP27. LY2228820 (200 nM–400 nM) enhances bortezomib-induced cytotoxicity and apoptosis, but LY2228820 alone doesn’t inhibit the growth of MM.1S cells. LY2228820 (200 nM–800 nM) also inhibits secretion of IL-6 and MIP-1α in long-term BM stromal cells (LT-BMSCs), BM mononuclear cells (BMMNCs), peripheral blood (PB) CD138⁺, CD138^? or PB CD14⁺ cells. LY2228820 (400 nM–800 nM) also blocks osteoclastogenesis from CD14⁺ cells. ^[2]

In Vivo

In LPS-induced mice, LY2228820 effectively inhibits the formation of TNFα with a threshold minimum 50% effective dose (TMED₅₀) less than 1 mg/kg. In a rat model of collagen-inducedarthritis (CIA), LY2228820 displays potent effects on paw swelling, bone erosion, and cartilage destruction, with a threshold minimum 50% effective dose (TMED₅₀)of 1.5 mg/kg. ^[1]

Clinical Trials

LY2228820 is currently under a Phase I clinical trial for the treatment of advanced cancer.

Features

Inhibition of p38α

Inhibition of p38α is determined using recombinant human p38α in a standard filter binding protocol using ATP[γ-³³P] and EGFR 21-mer peptide as substrate. Functional inhibition of TNFα in murine peritoneal macrophages is determined using LPS stimulation in the presence of LY2228820. To assess p38α activity in cells more directly, RAW 264.7 cells are treated with LY2228820 and then stimulated with anisomycin. The level of p38α activity is detected using a phosphoMAPKAPK-2 (pMK2) (Thr 334) antibody which reacts with a residue specifically phosphorylated by p38α.

Cell Lines

MM cells, including INA6, RPMI-8226, U266, and RPMI-Dox40

Concentrations

200 nM–800 nM

Incubation Time

48 hours

Methods

MTT assays and APO 2.7 staining are performed to assess cellular proliferation and induction of apoptosis, respectively. Viability is expressed as percent viable cells. Apoptosis in cells is evaluated by APO 2.7 staining. For detection of mitochondrial membrane protein 7A6 expressed in apoptotic cells, cells are incubated with APO 2.7 reagent for 20 min. Expression of APO 2.7 is determined using an EPICS XL flow cytometer.

Animal Models

Lipopolysaccharide (LPS)-induced Balb/c mice

Formulation

Dissolved in 1% CMC/0.25% Tween 80 in water

Doses

0–20 mg/kg

Administration

Oral bid dosing for 14 days