Cell experiment:

HUVECs or LECs (1200) are seeded into fibronectin-coated 96-well plates. After 24 hours, the cells are transferred into LEC medium containing 2% fetal bovine serum and incubated for an additional 24 hours. Cells (eight wells/condition) are incubated with medium alone (control), 20 ng/mL VEGF-A, or a combination of 20 ng/mL VEGF-A and 1 nM to 1 μM NVP-BAW2881. Proliferation is also assayed in LECs incubated with 500 ng/mL VEGF-C. The DMSO is adjusted to 0.1% in all wells. After 72 hours, cells are incubated with 5-methylumbelliferylheptanoate for subsequent fluorescent quantification of viable cells, using a electron microscope[2].

Animal experiment:

Mice: A contact hypersensitivity response is induced in the ear skin of 8-week-old female K14/VEGF-A TG mice. Five days after sensitization (day 0), the right ear is challenged by topical application of 10 μL oxazolone (1%) on each side. Starting on day 7, once-daily oral doses of 25 mg/kg NVP-BAW2881 or twice-daily topical doses of 0.5% NVP-BAW2881 are administered for 14 days. Control groups are given vehicles alone. The ear thickness is measured every other day using calipers. On day 21, mice are sacrificed and the weight of each ear and of its draining retro-auricular lymph node (LN) is determined[2].

IC50: 1.0-4.3 nM for VEGFR1-3

BAW2881 (NVP-BAW2881) is a VEGFR inhibitor.

Vascular endothelial growth factor (VEGF) is a potent endothelial cell mitogen regulating blood and lymphatic vessel development and homeostasis. There are three main subtypes of VEGFR, numbered VEGFR 1, 2 and 3.

In vitro: Previous study showed that BAW2881 could inhibit a limited number of kinases including c-RAF, B-RAF, RET, ABL, and TIE-2 at submicromole IC50s. BAW2881 could also inhibit the ligand induced autophosphorylation of RET, PDGFR, and KIT kinases. Moreover, low IC50 value (0.12 ± 0.06 nM) demonstrated that BAW2881 remarkably abrogated VEGF induced proliferation [1].

In vivo: In a psoriasis mouse model, BAW2881 was able to reduce the number of blood and lymphatic vessels and infiltrating leukocytes in the skin, and normalize the epidermal architecture. BAW2881 also showed strong anti-inflammatory effects in acute inflammation models. Moreover, the pretreatment with topical BAW2881 could significantly inhibit VEGF-A-induced vascular permeability in the skin of both pigs and mice. In addition, it was found that the topical application of BAW2881 was able to reduce the inflammatory response in pig skin caused by UV-B irradiation or by contact hypersensitivity reactions [2].

Clinical trial: Up to now, BAW2881 is still in the preclinical development stage.

References:
[1] Bold G et al.  A Novel Potent Oral Series of VEGFR2 Inhibitors Abrogate Tumor Growth by Inhibiting Angiogenesis. J Med Chem. 2016 Jan 14;59(1):132-46.
[2] Halin C,Fahrngruber H,Meingassner JG,Bold G,Littlewood-Evans A,Stuetz A,Detmar M.  Inhibition of chronic and acute skin inflammation by treatment with a vascular endothelial growth factor receptor tyrosine kinase inhibitor. Am J Pathol.2008 Jul;173(1):265-77.