ONX-0914 (PR-957)

Description

Biological Activity

Protocol(Only for Reference)

Chemical Information

Kinase Assay: [1]

Animal Study: [1]

References

貨號 Cat. No.: S7172
規格 Size: 5mgã€25mg
品牌 Brand: Selleckchem

ONX-0914 (PR-957)

Catalog No.S7172

ONX-0914 (PR-957) is a potent and selective immunoproteasome inhibitor with minimal cross-reactivity for the constitutive proteasome.

Size

Price

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Quantity

5mg	USD 185	In stock	0 1 2 3 4 5 6 7 8 9 10
25mg	USD 592	In stock	0 1 2 3 4 5 6 7 8 9 10

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

Proteasome active site ELISA

Briefly, samples (lysed cells or tissue homogenates) are treated for 1 hr at room temperature with the biotinylated active site probe PR-584 (5-15 μM). Samples are denatured by addition of SDS (0.9% final) and heating to 100°C for 5 minutes. The denatured samples are transferred to a 96-well or 384-well filter plate, mixed with streptavidin-sepharose beads (2.5-5 μL packed beads/well), and incubated for 1 hour at room temperature on a plate shaker. The beads are washed 5 times with 100-200 μL/well of ELISA buffer (PBS, 1% bovine serum albumin, 0.1% Tween-20) by vacuum filtration. The beads are incubated overnight at 4°C on a plate shaker with the following antibodies recognizing the six catalytic subunits diluted into ELISA buffer: β5, β1, and β2 diluted 1:3000; LMP7 and LMP2 diluted 1:5000; and MECL-1 diluted 1:1000. The beads are washed 5 times with 100-200 μL/well of ELISA buffer and incubated with HRP-conjugated secondary antibody (goat anti-rabbit for β5, rabbit anti-goat for MECL-1, and goat anti-mouse for LMP7, LMP2, β1 and β2) diluted 1:5000 in ELISA buffer and incubated 2 hours at room temperature on a plate shaker. The beads are washed 5 times with 100-200 μL/well of ELISA buffer and developed for chemiluminsecence signal using the supersignal ELISA pico substrate following the manufacturer’s instructions. Luminescence is measured on a plate reader and converted to ng of proteasome or μg/mL of lysate by comparison with 20S proteasome or untreated cell lysate standard curves.

Animal Models	collagen antibody–induced arthritis (CAIA) and collagen-induced arthritis (CIA)
Dosages	2, 6 or 10 mg/kg
Administration	i.v.

Description

ONX-0914 (PR-957) is a potent and selective immunoproteasome inhibitor with minimal cross-reactivity for the constitutive proteasome.

In vitro

ONX-0914 is 20- to 40-fold more selective for LMP7 over the next most sensitive sites, β5 or LMP2. ONX-0914 blocks presentation of LMP7-specific, MHC-I–restricted antigens in vitro and in vivo with minimal cross-reactivity for the constitutive proteasome. Selective inhibition of LMP7 by ONX-0914 blocks production of interleukin-23 (IL-23) by activated monocytes and interferon-gamma and IL-2 by T cells. LMP7 inhibition blocks production of IL-23 by ~90% and of tumor necrosis factor-α (TNF-α) and IL-6 by ~50%.[1]

In mouse models of rheumatoid arthritis and lupus, ONX-0914 treatment reverses signs of disease and results in reductions in cellular infiltration, cytokine production and autoantibody levels at well-tolerated doses. The maximum tolerated dose (MTD) of ONX-0914 in mice to be 30 mg/kg body weight. IFN-g release is inhibited by ~60% at LMP7-selective concentrations of ONX-0914 and by ~90% at higher concentrations. Production of IL-2 is also inhibited by ~50%.[1]

The first highly selective, small molecule inhibitor of the immunoproteasome. Potential use in cancer and autoimmune diseases (e.g. rheumatoid arthritis, inflammatory bowel disease, and lupus).

Download ONX-0914 (PR-957) SDF

Molecular Weight (MW)	580.67
Formula	C31H40N4O7
CAS No.	960374-59-8

Storage	3 years -20â„ƒPowder
Storage	6 months-80â„ƒin DMSO
Syonnyms	N/A

Solubility (25°C) *	In vitro	DMSO	100 mg/mL (172 mM)
		Water	100 mg/mL (172 mM)
		Ethanol	100 mg/mL (172 mM)
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.