Pan-Caspase inhibitor - Z-VAD-FMK

Caspase-1 inhibitor

Inhibition of caspase signaling by Z-VAD-FMK

Z-VAD-FMK is a cell-permeable pan-caspase inhibitor [1, 2]. It potently inhibits human caspase-1 to -10 with the exception of caspase-2 [3]. It also inhibits murine caspases, notably caspase-1, caspase-3, and caspase-11, the ortholog of human caspase-4 and -5 [4, 5]. Caspases are a family of cysteine proteases that are centrally involved in cell death and inflammation responses [6, 7].

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Mode of action:

Z-VAD-FMK inhibits caspases by irreversibly binding to their catalytic site [1]. By inhibiting the activity of multiple caspases, Z-VAD-FMK can block many different biological processes including inflammasome activation and the induction of apoptosis leading to increased cell survival in many different cell types [2-5]. Interestingly, It has been shown that Z-VAD‑FMK administration can significantly reduce inflammation and lethality in an experimental model of endotoxic shock [5]. To conclude, this broad-spectrum inhibitor is a useful tool for studying the role of caspases in inflammation and cell death.

Key features:

• Broad-spectrum caspase inhibitor
• Potent inhibitor of caspase-dependent inflammasomes
• Blocks the induction of apoptosis
• InvitroFit™ grade: each lot is highly pure (≥95%) and functionally tested

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References:

1. Slee EA. et al., 1996. Benzyloxycarbonyl-Val-Ala-Asp (OMe) fluoromethylketone (Z-VAD.FMK) inhibits apoptosis by blocking the processing of CPP32. Biochem J. 315 ( Pt 1):21-4.
2. Li X. et al., 2019. The caspase inhibitor Z-VAD-FMK alleviates endotoxic shock via inducing macrophages necroptosis and promoting MDSCs-mediated inhibition of macrophages activation. Front Immunol. 10:1824.
3. Chauvier D. et al., 2007. Broad-spectrum caspase inhibitors: from myth to reality? Cell Death Differ. 14:387-91.
4. Guey B. et al., 2014. Caspase-1 autoproteolysis is differentially required for NLRP1b and NLRP3 inflammasome function. PNAS 11(48):17254-9.
5. Py B.F. et al., 2014. Caspase-11 controls interleukin-1β release through degradation of TRPC1. Cell Rep. 6: 1122–8.
6. Van Opdenbosch N. & Lamkanfi M. et al., 2019. Caspases in Cell Death, Inflammation, and Disease. Immunity. 50(6):1352-1364.
7. Tsuchiya K. et al., 2019. Caspase-1 initiates apoptosis in the absence of gasdermin D. Nat Commun. 10(1):2091.

Figures

Z-VAD-FMK inhibits the NLRP3 inflammasome response in a dose-dependent manner.
THP1-Null2 cells, primed with LPS‑EK (1 μg/ml for 3 h), were stimulated with MSU crystals (150 µg/ml) and increasing concentrations of Z-VAD‑FMK. After overnight incubation, IL-1β secretion was analyzed by adding 50 µl of supernatant from treated THP1-Null2 cells 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 QUANTI‑Blue™ Solution, a SEAP detection reagent, and by reading the optical density (OD) at 655 nm. Data are shown as percentage (%) inhibition.

Specifications

Working concentration: 10 μg/ml (20 μM)

Synonym: Carbobenzoxy-valyl-alanyl-aspartyl-[O- methyl]- fluoromethylketone

Solubility: Soluble in DMSO (10 mg/ml)

Molecular weight: 467.5 g/mol

Quality control:

• Purity: ≥95% (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

• 1 mg of Z-VAD-FMK (provided as a translucent film)

Z-VAD-FMK 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

Caspases in inflammation & cell death:

The caspase enzymes are a family of cytosolic proteases involved in the regulation of inflammation and cell death [1, 2]. They can be divided into inflammatory caspases (such as caspases-1, -4, -5, and -12 in humans and caspases-1, -11, and -12 in mice) and apoptotic caspases (such as caspases-2, -3, -6, -7, -8, -9, and -10) [3].

Notably, caspase-1 plays a crucial role in the control of inflammation. Its activity is regulated by a multi-protein complex known as the inflammasome, inflammasome activation drives oligomerization and self-activation of caspase-1. Upon activation, caspase-1 processes the inflammatory cytokines, interleukin-1β (IL-1β) and IL-18, and Gasdermin-D, promoting inflammation and pyroptosis, a form of cell death.

1. Van Opdenbosch N. & Lamkanfi M. et al., 2019. Caspases in Cell Death, Inflammation, and Disease. Immunity. 50(6):1352-1364.
2. Tsuchiya K.et al., 2019. Caspase-1 initiates apoptosis in the absence of gasdermin D. Nat Commun. 10(1):2091.
3. Shalini M. et al., 2015. Old, new and emerging functions of caspases. Cell Death Differ. 22:526-39.

Chemical structure of Z-VAD-FMK: