2-Iminobiotin hydrobromide

SKU: orb1299979

Description

2-Iminobiotin hydrobromide is a cyclic guanidino derivative of biotin which functions as a reversible, competitive inhibitor of both inducible (iNOS) and neuronal (nNOS) nitric oxide synthase isoforms. This small molecule is utilized in vitro and in vivo to investigate the pathophysiological roles of NO overproduction in models of inflammation, sepsis, and neurological disorders.

Research Area

Immunology & Inflammation

Key Properties

• CAS Number: 76985-52-9
• MW: 324.24
• Purity: 98.13%
• Formula: C₁₀H₁₈BrN₃O₂S
• SMILES: [H][C@](CS[C@H]1CCCCC(O)=O)([C@]1([H])N2)NC2=N.Br
• Target: NO Synthase,NOS
• Solubility: DMSO:12.5 mg/mL (38.55 mM);10% DMSO+40% PEG300+5% Tween 80+45% Saline:2 mg/mL (6.17 mM)

Bioactivity

• Target IC50:
  iNOS:21.8 µM(Ki) (mouse)|nNOS:37.5 µM(Ki)(rat)
• In Vitro:
  All cultures were subjected to 25 h of hypothermia (33.5°C), and incubated with vehicle or 2-iminobiotin (2-IB) (10, 30, 50, 100, and 300 ng/ml). Cell morphology was evaluated by brightfield microscopy. Cell damage was analyzed by LDH assays. Production of reactive oxygen species (ROS) was measured using fluorometric assays. Western blotting for PARP, Caspase-3, and the phosphorylated forms of akt and erk1/2 was conducted. To evaluate early apoptotic events and signaling, cell protein was isolated 4 h post-hypoxia and human apoptosis proteome profiler arrays were performed. Twenty-five hour after the hypoxic insult, clear morphological signs of cell damage were visible and significant LDH release as well as ROS production were observed even under hypothermic conditions. Post-hypoxic application of 2-IB (10 and 30 ng/ml) reduced the hypoxia-induced LDH release but not ROS production. Phosphorylation of erk1/2 was significantly increased after hypoxia, while phosphorylation of akt, protein expression of Caspase-3 and cleavage of PARP were only slightly increased. Addition of 2-IB did not affect any of the investigated proteins. Apoptosis proteome profiler arrays performed with cellular protein obtained 4 h after hypoxia revealed that post-hypoxic application of 2-IB resulted in a ≥ 25% down regulation of 10/35 apoptosis-related proteins: Bad, Bax, Bcl-2, cleaved Caspase-3, TRAILR1, TRAILR2, PON2, p21, p27, and phospho Rad17.
• Cell Research:
  In vitro hypoxia was induced for 7 h in IMR-32 cell cultures by using our recently described system with minor modifications. Enzyme stock solutions (100x) of catalase and glucose oxidase were diluted in cell culture medium (DMEM/F12, 1% FCS; final concentration: 120 and 2 U/ml respectively). A rapid decrease of partial pressure of oxygen (pO2) to levels below 10 mmHg was achieved by adding the enzymes to glucose containing culture medium. Also a decline in glucose (<1 g/l) and pH (<7.0) was observed, resembling the clinical characteristics of hypoxic-ischemic injury in vivo. Hypoxic conditions were confirmed with a tissue oxygen pressure monitor . After the hypoxic insult, cells were washed twice with PBS and cultures were placed into an incubator at 33.5°C (hypothermia) employing culture medium with (i) solvent (citrate buffer 1%) or (ii) 2-IB at 10, 30, 50, 100, and 300 ng/ml. To determine the optimal “reperfusion” time, a time-interval curve investigating cell damage (LDH release) was performed. Analyses of LDH release, ROS generation, hydrogen peroxide release, metabolic activity, cell signaling, apoptosis-related protein expression/activity and expression analysis of 35 human apoptosis-related proteins were performed at different time points post-hypoxia

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

2 Iminobiotin hydrobromide, 2-Iminobiotin, 2Iminobiotin hydrobromide, 2-Iminobiotin Hydrobromide, Guanidinobiotin, Inhibitor, iNOS, NOS, NOSynthase, inhibit, nNOS, NO Synthase, Nitric oxide synthases

Compound Identifiers

PubChem CID

131667834