| Size | Price | Stock | Qty |
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| Other Sizes |
Purity: ≥98%
| Targets |
Gasdermin D[1].
LDC7559 targets gasdermin D (GSDMD), a key effector protein in the pyroptosis pathway. Pyroptosis is a form of programmed cell death that is triggered by inflammasome activation and characterized by the release of pro-inflammatory cytokines such as IL-1β. GSDMD is cleaved by inflammatory caspases, generating an N-terminal fragment that forms pores in the plasma membrane, leading to cell lysis and cytokine release. LDC7559 inhibits GSDMD by blocking NET formation at a late stage. |
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| ln Vitro |
LDC7559 (1 and 5 μM) dramatically reduces the lethal effect of human and mouse GSDMD NT domains transfected into HEK293T cells and decreases the release of IL-1β upon inflammasome activation [3]. Rather than interfering with the activation and cleavage mechanism, LDC7559 acts directly by preventing the GSDMD NT domain from functioning [3].
In vitro, LDC7559 (1 and 5 μM) inhibits IL-1β release upon inflammasome activation and significantly blocks the lethal effect of both human and murine GSDMD NT domains transfected into HEK293T cells. It also decreases the release of IL-1β upon inflammasome activation. These in vitro activities support its use in studying pyroptosis, inflammasome signaling, and inflammatory diseases. |
| ln Vivo |
In vivo data for LDC7559 is not extensively reported in publicly available sources. As a GSDMD inhibitor that blocks pyroptosis and reduces inflammation, the compound has potential applications in animal models of inflammatory diseases, sepsis, and autoimmune disorders. However, specific published in vivo efficacy studies are not detailed in the current literature. LDC7559 is primarily used as a research tool for studying pyroptosis and gasdermin D biology.
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| Enzyme Assay |
The in vitro GSDMD inhibition assay for LDC7559 uses HEK293T cells transfected with human or murine GSDMD NT domains. Cells are treated with LDC7559 at concentrations such as 1 and 5 μM, and cell death is measured using standard viability assays such as LDH release or propidium iodide staining. IL-1β release is measured by ELISA in inflammasome-activated cells. NET formation is assessed in neutrophils using fluorescence microscopy.
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| Cell Assay |
Transfection of GSDMD constructs in HEK293T cells [2]
HEK293T cells were seeded the day before transfection at a density of 5 × 105 cells/ml. Transfection was done in Opti-MEM using 2 μg of DNA and 7 μl of Lipofectamine 2000 for a six-well plate or respective scaling for other surface areas. LDC7559 or caspase inhibitors were added 2 hours after transfection, and lactate dehydrogenase (LDH) release of cells was measured 16 hours after transfection. Incubation of HEK293T lysates with recombinant proteases[2] HEK293T cell lysate was harvested 16 hours after transfection by changing the medium of a six well to 200 μl of Opti-MEM containing 0.05% (v/v) Triton X-100 and scraping cells off the bottom of the well. After 10 min of centrifugation at full speed and 4°C in a tabletop centrifuge, supernatants were used for cleavage assays. Fifty microliters of lysate were incubated with 10 mU of purified human NE, 2 U of recombinant caspase-4, 4 μg of purified CG (RP-77525), or 6.25 μg of recombinant PR3 (C628) for the indicated time points at 25°C on a shaking incubator. Reactions were stopped by directly adding 50 μl of 2× SDS loading buffer (at 98°C) containing 200 mM DTT and by immediately boiling samples before loading on SDS–polyacrylamide gel electrophoresis gels. Cellular assays for LDC7559 are conducted in HEK293T cells expressing GSDMD NT domains and in inflammasome-activated immune cells. Cells are treated with varying concentrations of LDC7559. Cell death is assessed by LDH release or propidium iodide staining. IL-1β release is measured by ELISA. NET formation is evaluated in neutrophils using fluorescence microscopy. The compound's effects on pyroptosis and inflammation are evaluated by measuring cytokine levels and cell lysis. |
| Animal Protocol |
Live imaging of neutrophils [2]
Cells were resuspended in an assay medium supplemented with 500 nM SYTOX Green and 2.5 μM DRAQ5 and seeded at a density of 5 × 105 cells per well into μ-Slide 8 Well ibiTreat dishes (ibidi). After LDC7559 treatment and NET induction, images were collected at 2-min intervals on a Leica TCS SP8 confocal microscope equipped with a climate chamber at 37°C and with a Leica HC PL APO 20×/0.75 IMM CORR CS2 objective using glycerol immersion. The recording of DRAQ5 (cell-permeable DNA dye) was used to track individual nuclei over time and to determine nuclear area. The recording of SYTOX Green (cell-impermeable dye) was used to determine permeability of cells. Bright-field recording was added for a video representation of the data. Automated algorithms were used to determine the time point of permeabilization and to categorize nuclei into “non-expanded,” “fully expanded” (filling the entire cell), or “NET” (extending beyond the maximal area of a cell). The results were then mapped back to a video representation of the data and manually inspected for accuracy before they were used for further analysis. The complete image analysis pipeline consisting of ImageJ and R scripts will be described elsewhere in detail. In vivo studies for LDC7559 would typically involve animal models of sepsis, inflammatory diseases, or autoimmune disorders. The compound would be administered via intraperitoneal or oral routes at doses determined by pharmacokinetic studies. Efficacy would be assessed by measuring inflammatory cytokine levels, tissue damage, and disease severity. However, specific published in vivo protocols for LDC7559 are not available in the current literature. The compound is currently used as a research tool. |
| ADME/Pharmacokinetics |
Pharmacokinetic data for LDC7559 is not extensively reported in publicly available sources. The compound has a molecular weight of 349.38 g/mol and a molecular formula of C20H19N3O3. It has a CAS number of 2407782-01-6. It is soluble in DMSO at 95 mg/mL. Storage: powder at -20°C for 3 years. As a small molecule, it is expected to have moderate bioavailability. Detailed PK parameters such as half-life are not available.
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| Toxicity/Toxicokinetics |
Toxicity data for LDC7559 is limited in publicly available sources. As with all research compounds, LDC7559 is intended for research use only and not for human therapeutic applications. Standard in vitro cytotoxicity assays and in vivo tolerability studies would be required for a complete toxicity assessment. The compound's mechanism of inhibiting pyroptosis suggests potential immunomodulatory effects that would need to be evaluated.
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| References | |
| Additional Infomation |
LDC7559 (CAS 2407782-01-6) is a potent gasdermin D (GSDMD) inhibitor that blocks NET formation at a late stage. It inhibits IL-1β release upon inflammasome activation and blocks the lethal effect of GSDMD NT domains. LDC7559 has a molecular formula of C20H19N3O3 and a molecular weight of 349.38 g/mol. It is a valuable research tool for studying pyroptosis, inflammasome signaling, and inflammatory diseases.
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| Molecular Formula |
C20H19N3O3
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|---|---|
| Molecular Weight |
349.38
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| Exact Mass |
349.142
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| Elemental Analysis |
C, 68.75; H, 5.48; N, 12.03; O, 13.74
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| CAS # |
2407782-01-6
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| PubChem CID |
138319675
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| Appearance |
White to light yellow solid
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
596.3±50.0 °C at 760 mmHg
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| Flash Point |
314.4±30.1 °C
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| Vapour Pressure |
0.0±1.7 mmHg at 25°C
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| Index of Refraction |
1.647
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| LogP |
2.25
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
26
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| Complexity |
503
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O1C2C=C(C=CC=2CN2C(=CC(C3C=CC=CC=3OC)=N2)C1)NC(C)=O
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| InChi Key |
IFOIINXSFCQLOV-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C20H19N3O3/c1-13(24)21-15-8-7-14-11-23-16(12-26-20(14)9-15)10-18(22-23)17-5-3-4-6-19(17)25-2/h3-10H,11-12H2,1-2H3,(H,21,24)
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| Chemical Name |
N-(2-(2-Methoxyphenyl)-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)acetamide
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : 100 mg/mL (286.22 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (5.95 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (5.95 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (5.95 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.8622 mL | 14.3111 mL | 28.6221 mL | |
| 5 mM | 0.5724 mL | 2.8622 mL | 5.7244 mL | |
| 10 mM | 0.2862 mL | 1.4311 mL | 2.8622 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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