CWHM-12

SKU: orb1296100

Description

CWHM-12

Research Area

Signal Transduction

Key Properties

• CAS Number: 1564286-55-0
• MW: 590.47
• Purity: 99.83%
• Formula: C₂₆H₃₂BrN₅O₆
• SMILES: CC(C)(C)c1cc(Br)cc(c1)[C@H](CC(O)=O)NC(=O)CNC(=O)c1cc(O)cc(NC2=NCC(O)CN2)c1
• Target: Integrin
• Solubility: 10% DMSO+40% PEG300+5% Tween 80+45% Saline:2 mg/mL (3.39 mM);DMSO:45 mg/mL (76.21 mM)

Bioactivity

• Target IC50:
  αvβ1 integrin:1.8 nM|αvβ8 integrin:0.2 nM|αvβ3 integrin:0.8 nM|αvβ6 integrin:1.5 nM|αvβ5 integrin:61 nM
• In Vivo:
  Mice are treated with CCl4 for 3 weeks to establish fibrotic disease and then treated with CWHM-12 or vehicle for the final 3 weeks of CCl4. CWHM-12 significantly reduces liver fibrosis even after the fibrotic disease has been established. Digital image quantitation demonstrates significantly reduced p-SMAD3 signaling in the livers of CWHM-12 treated mice compared to controls, demonstrating that the protection from CCl4-induced hepatic fibrosis observed in CWHM-12 treated mice is due at least in part to a reduction in TGF-β activation by αv integrins. Besides, the administration of CWHM-12 significantly inhibited the progression of pulmonary fibrosis.
• In Vitro:
  CWHM-12 also less potently inhibits αvβ5 (IC50: 61 nM) and αIIbβ3/α2β1/α10β1 (IC50>5000 nM). CWHM-12 demonstrates high potency against all of the five possible β subunit binding partners (αvβ1, αvβ3, αvβ5, αvβ6, and αvβ8) in in vitro ligand-binding assays, with somewhat less potency against αvβ5 than against the other αv integrins.
• Cell Research:
  The stably transfected human 293 cells over-expressing human αvβ3 or αvβ5 are pre-incubated in HBSS buffer containing 200 μM MnCl2 for 30 min at 37°C with 3-fold dilutions of compound (e.g., CWHM-12). Each sample is then added to triplicate wells of a 96-well plate which has been coated overnight at 4°C with a predetermined optimal concentration of purified vitronectin, washed, blocked by 1 hr incubation with BSA, and washed again. Cells are allowed to attach for 30 min at 37°C, and non-adherent cells are removed by washing. Remaining attached cells are measured by endogenous alkaline phosphatase activity using para-nitrophenyl phosphate and reading absorbance signal at 405 nM. The same procedure is used to measure adhesion of αvβ6-expressing human HT-29 cells to purified human latency-associated peptide, and α5β1-expressing human K562 cells to human plasma fibronectin. In all cell-based assays, binding by the expected integrin is verified by the testing activity of corresponding isotype-matched positive (function-blocking) and negative control antibodies.
• Animal Research:
  The mTmG (Td tomato/EGFP) and Ai14 (Rosa-CAG-LSL-tdTomato-WPRE) mice are used and crossed with Pdgfrb-Cre mice. Wild type C57/BL6 mice, Itgavflox/flox mice, and itgb8flox/flox mice are used. Mice used for all experiments are 8-12 weeks old and are housed under specific pathogen-free conditions. For all studies, CWHM-12 and CWHM-96 are solubilized in 50% DMSO (in sterile water) and dosed to 100 mg/kg/day. Drug or vehicle (50% DMSO) is delivered by implantable ALZET osmotic minipumps. For CCl4-induced fibrosis, pumps are inserted subcutaneously either before the first dose of CCl4 (prophylactic) or after 3 weeks of treatment (therapeutic) and livers are harvested after 6 weeks. For Bleomycin-induced fibrosis pumps are inserted 14 days after treatment with Bleomycin or saline and lungs are harvested at 28 days.

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

CWHM 12, CWHM12, CWHM-12, inhibit, Inhibitor, Integrin, αVβ3 integrin, αVβ6 integrin, αVβ8 integrin, αVβ1 integrin

Compound Identifiers

PubChem CID

72949858