The Pyrazolo[3,4-d]pyrimidine-Based Kinase Inhibitor NVP-BHG712: Effects of Regioisomers on Tumor Growth, Perfusion, and Hypoxia in EphB4-Positive A375 Melanoma Xenografts

Abstract

In a previous study, EphB4 was demonstrated to be a positive regulator of A375melanoma growth but a negative regulator of tumor vascularization and perfusion. To distinguish between EphB4 forward and ephrinB2 reverse signaling, we used the commercially available EphB4 kinase inhibitor NVPBHG712 (NVP), which was later identified as its regioisomer NVPiso. Since there have been reported significant differences between the inhibition profiles of NVP and NVPiso, we compared the influence of NVP and NVPiso on tumor characteristics under the same experimental conditions. Despite the different inhibitory profiles of NVP and NVPiso, the comparative study conducted here showed the same EphB4-induced effects in vivo as in the previous investigation. This confirmed the conclusion that EphB4-ephrinB2 reverse signaling is responsible for increased tumor growth as well as decreased tumor vascularization and perfusion. These results are further substantiated by microarrays showing differences between mock-transfected and EphB4-transfected (A375-EphB4) cells with respect to at least 9 angiogenesis-related proteins. Decreased expression of vascular endothelial growth factor (VEGF), angiotensin 1 (Ang-1), and protein kinase B (Akt/PKB), together with the increased expression of tissue inhibitor of metalloproteinase-1 (TIMP-1) and transforming growth factor beta-2 (TGF-β2), is consistent with the impaired vascularization of A375-EphB4 xenografts. Functional overexpression of EphB4 in A375-EphB4 cells was confirmed by activation of a variety of signaling pathways, including the Janus kinase/signal transducers and activators of transcription (JAK/STAT), rat sarcoma virus/rapidly accelerated fibrosarcoma/mitogen activated protein kinase kinase (Ras/Raf/MEK), and nuclear factor kappa-B (NFkB) pathways.

Robust identification of shared key genomic biomarkers for diagnosis and therapies of pancreatic cancer with type-2 diabetes

“…Linsitinib, an investigational cancer drug (DB accession number DB06075), inhibits insulin-like growth factor receptor-1 and has diverse effects on PC growth and metastasis [13,99]. NVP-BHG712, a kinase inhibitor, has implications in blood circulation, oxygen de ciency, and cancer progression [100]. It is also a speci c inhibitor of EphB4 kinase, which may be a therapeutic target for insulin resistance and T2D [101,102].…”

Immune escape and metastasis mechanisms in melanoma: breaking down the dichotomy

“…This effectively created a hypoxic environment, especially in small tumors. EPHB4 also increased expression of the immunosuppressant IL-8 and TGF-β2 ( 152 , 153 ). In addition, TGF-β2 was shown to act as a master regulator of EMT in acidosis adapted cells, in colorectal and squamous cell carcinomas ( 154 ).…”