GSK484 hydrochloride

SKU: orb1296229

Description

GSK484 hydrochloride is a potent, reversible PAD4 inhibitor with IC50 values of 50 nM (without Ca²⁺) and 250 nM (with 2 mM Ca²⁺). It has been shown to enhance the radiosensitivity of colorectal cancer cells and to suppress neutrophil extracellular trap (NET) formation in both cellular and animal studies.

Research Area

Epigenetics & Chromatin

Key Properties

• CAS Number: 1652591-81-5
• MW: 510.03
• Purity: 98.32%
• Formula: C₂₇H₃₂ClN₅O₃
• SMILES: Cl.COc1cc(cc2nc(-c3cc4ccccc4n3CC3CC3)n(C)c12)C(=O)N1CC[C@@H](O)[C@@H](N)C1
• Target: PAD
• Solubility: PBS:<1 mg/mL (insoluble or slightly soluble);DMSO:247.5 mg/mL (485.27 mM);H2O:100 mg/mL (196.07 mM)

Bioactivity

• Target IC50:
  PAD4:250 nM (the presence of 2 mM Calcium)|PAD4:50 nM (0 mM Calcium)|PAD4:50 nM (2 mM Calcium)
• In Vivo:
  To determine if PAD4 inhibition can mitigate cancer-associated kidney damage, MMTV-PyMT mice received the PAD4 inhibitor GSK484 at a dosage of 4 mg/kg daily for one week. This treatment effectively lowered the elevated neutrophil NETosis in the peripheral blood of cancerous mice. Concurrently, there was a significant reduction in the urine protein level of treated MMTV-PyMT mice compared to those untreated, indicating kidney function improvement akin to that achieved with DNase I treatment, without any observed toxicity .
• In Vitro:
  METHODS: GSK484 hydrochloride (GTPL8577) (10 μM) treated mouse and human neutrophils, demonstrating inhibition of citrullination by Western blot and novel imaging assays in neutrophils stimulated with calcium ionophore or bacteria and further Study the effects on NET formation in vitro by mouse and human neutrophils. RESULTS GSK484 hydrochloride pretreatment significantly reduced citrulline production and NET formation.
• Cell Research:
  HEK293 cells stably expressing N-terminal FLAG-tagged PAD1, PAD2, PAD3 or PAD4 are engineered by retroviral transduction. Cells are grown in 15 cm diameter plates to subconfluency in DMEM supplemented with 10% Foetal Bovine Serum, harvested by centrifugation and washed once in PBS/2 mM EGTA. Cells are lysed in 50 mM Tris-Cl, pH 7.4, 1.5 mM MgCl2, 5% glycerol, 150 mM NaCl, 25 mM NaF, 1 mM Na3VO4, 0.4% NP40, 1 mM DTT with protease inhibitors. Lysates are pre-incubated for 20 min at 4°C with DMSO alone (2%), 100 μM of GSK199, GSK484, GSK106 or 200 μM Cl-amidine. Citrullination reactions are performed for 30 min at 37°C in the presence of 2 mM calcium. Extracts are loaded on to gels, proteins separated by SDS-PAGE and transferred to PVDF membranes. Citrullinated proteins are then chemically modified and detected using an anti-modified citrulline antibody. FLAG-PAD constructs are detected using the anti-FLAG antibody .
• Animal Research:
  The study includes the MMTV-PyMT mouse model for mammary carcinoma (FVB/n background) and the RIP1-Tag2 mouse model for pancreatic neuroendocrine carcinoma (C57BL/6 background). Mice are treated daily by intra-peritoneal injections of the PAD4 inhibitor GSK484 (4 mg/kg). GSK484 is dissolved in 99.9% ethanol at a concentration of 25 mg/mL to generate a stock solution and further diluted 1:50 in 0.9% NaCl shortly before injection of 200 μL/mouse .

Storage & Handling

• Storage: Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice/Shipping at ambient temperature.
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

Inhibitor, GTPL 8577, GTPL8577, GTPL-8577, GSK484, GSK-484, GSK484 hydrochloride, GSK484 Hydrochloride, GSK-484 hydrochloride, GSK-484 Hydrochloride, GSK 484, GSK 484 Hydrochloride, inhibit, PAD4, AOB 6992, AOB6992, AOB-6992, Protein Arginine Deiminase, Peptidylarginine Deiminase

Compound Identifiers

PubChem CID

86340151