Caspase-1 and Caspase-4 inhibitor

Caspase-1 and Caspase-4 inhibitor

Casp-1 and Casp-4 inhibition by VX-765

VX-765 is a pro-drug converted by plasma esterases into an active peptidomimetic metabolite, VRT-043198, which potently inhibits caspase-1 and caspase-4 [1].

CASPASE-1 & CASPASE-4:

Both caspase-1 and caspase-4 belong to a family of inflammatory caspases that are crucial in regulating cell death and inflammation. Notably, inflammasome induction drives the self-cleavage and activation of caspase-1 which in turn leads to cleavage of the pro‑inflammatory cytokines interleukin-1 beta (IL-1β)/IL-18 and the pore-forming protein Gasdermin D (GSDMD) into their active forms. Additionally, the activation of the inflammasome also leads to alarmin secretion and pyroptosis, a form of immunogenic cell death.

MODE OF ACTION OF VX-765:

VX-765 acts by covalent modification of the catalytic cysteine residue in the active site of caspase-1. Through its inhibitory activity, it has been demonstrated that VX-765 reduces the production of IL-1β and IL-18 in models of inflammatory disease both in vitro and in vivo [1, 2]. In addition, it has been reported that this inhibitor blocks pyroptosis [3].

KEY FEATURES OF VX-765 :

• Upon conversion, a potent inhibitor of caspase-1 and caspase-4
• Each lot is highly pure (≥97%) and functionally tested

Read our review on inflammasomes.

References:

1. Wannamaker W. et al., 2007. (S)-1-((S)-2-{[1-(4-amino-3-chloro-phenyl)-methanoyl]-amino}-3,3-dimethyl-butanoyl)-pyrrolidine-2-carboxylic acid ((2R,3S)-2-ethoxy-5-oxo-tetrahydro-furan-3-yl)-amide (VX-765), an orally available selective interleukin (IL)-converting enzyme/caspase-1 inhibitor, exhibits potent anti-inflammatory activities by inhibiting the release of IL-1beta and IL-18. J Pharmacol Exp Ther. 321(2):509-16.
2. McKenzie B.A. et al., 2018. Caspase-1 inhibition prevents glial inflammasome activation and pyroptosis in models of multiple sclerosis. PNAS. 115(26):E6065-E6074.
3. Doitsh G. et al., 2014. Cell death by pyroptosis drives CD4 T-cell depletion in HIV-1 infection. Nature. 505(7484):509-14.

Figures

IL-1β production in THP1 cells following their stimulation with CPPD, Alhydrogel and Chitosan
Cells treated or not with VX-765 (100µM) and primed with LPS (1 μg/ml) were stimulated with CPPD (50µg/ml),  Alhydrogel (1/100) and Chitosan (100µg/ml). After 24h incubation, the supernatants were added to HEK-Blue™ IL-1β cells. IL-1β-induced activation of NF-κB was assessed by measuring the levels of SEAP in the supernatant of HEK-Blue™ IL-1β cells using the QUANTI-Blue™ assay.

Lucia production  in THP1-Lucia™ ISG cells following their stimulation with human IFNα2a, 2' 3'-cGAMP and Poly(dA:dT) / LyoVec™.
Cells treated or not with VX-765 (100µM) were stimulated with IFNα2a (30UI/ml),  2' 3'-cGAMP (20µg/ml) and Poly(dA:dT) / LyoVec™ (500ng/ml). After 24h incubation, the levels of Lucia in the supernatant of THP1-Lucia™ ISG cells using the QUANTI-Luc™ assay.

Seap production  in THP1-Dual cells following their stimulation with LPS-EK (10ng/ml), 2' 3'-cGAMP (10µg/ml) and Pam3CSK4 (100 ng/ml).
After 24h incubation, the levels of SEAP in the supernatant of THP1-Dual cells is determined using the QUANTI-Blue™ assay.

Specifications

CAS number: 273404-37-8

Chemical Name: (S)-1-((S)-2-{[1-(4-amino-3-chloro-phenyl)-methanoyl]-amino}-3,3-dimethyl-butanoyl)-pyrrolidine-2-carboxylic acid ((2R,3S)-2-ethoxy-5-oxo-tetrahydro-furan-3-yl)-amide

Solubility: 100 mg/ml (200 mM) in DMSO or ethanol

Formula: C₂₄H₃₃ClN₄O₆

Molecular weight: 509 g/mol

Quality control

• Purity: ≥97% (UHPLC)
• The inhibitory activity of the product has been validated in inflammasome cellular assays using THP1-Null2 and HEK-Blue™ IL-1β cells.
• The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells.

Contents

VX-765 (provided as a translucent film) is available in 2 quantities:

• 10 mg VX-765 (#inh-vx765-1)
• 5 x 10 mg VX-765 (#inh-vx765-5)

VX-765 is shipped at room temperature.

Upon receipt, store at -20°C.

Resuspended product is stable for at least 6 months when properly stored.

Avoid repeated freeze-thaw cycles.

Details

Chemical structure of VX-765: