Crystal structure of the kinase domain of human vascular endothelial growth factor receptor 2: a key enzyme in angiogenesis

McTigue, Michele; Wickersham, John; Pinko, Christopher; Showalter, Richard E.; Parast, Camran V.; Tempczyk-Russell, Anna; Gehring, Michael R.; Mroczkowski, Barbara; Kan, Chen‐Chen; Villafranca, J. E.; Appelt, K.

doi:10.1016/s0969-2126(99)80042-2

Cited by 188 publications

(168 citation statements)

References 47 publications

(69 reference statements)

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“…The role of JM rearrangement in VEGFR2 activation can be inferred from the conformations of the structures reported here as well as the previously reported structure of the activated catalytic domain (21) (Fig. 3).…”

Section: Resultssupporting

confidence: 73%

“…1A). This cleft will be referred to here as the regulatory domain pocket (RDP), because it is also where DFG resides in the active conformation (DFG in ) and therefore, precludes a DFG in /JM in arrangement (21). Just before the classical kinase domain, the plus-JM residues 820-826 extend over the α-helix C. The density for residues 813-819 was not wellordered, and therefore, positions for these residues are not included in the crystal structure coordinates.…”

Section: Resultsmentioning

confidence: 99%

“…For example, protein constructs of VEGFR2 TK that contain only the catalytic domain but not an important regulatory domain have been broadly used as practical surrogates of full-length protein to determine biochemical potency. Additionally, the reported inhibitor VEGFR2 TK crystal structures contain constructs comprised of only the catalytic domain (21,25). Although these kinase catalytic domain constructs circumvent key technical problems inherent with full-length protein, the excised JM domain has been shown to impact both kinase regulation and measured inhibitor potency within the PDGF family of RTKs (14), including VEGFR2 TK (15).…”

Section: Discussionmentioning

confidence: 99%

“…The VEGFR TKIs that uniformly exhibit greater JM construct potency do not fill the RDP with a large inhibitor substituent, as type II TKIs do. The solved structures of these TKIs reveal a stabilized JM in acting as the physical lock in the RDP, precluding the DFG loop from adopting an active in position (21) (Fig. 3).…”

Section: Discussionmentioning

confidence: 99%

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Molecular conformations, interactions, and properties associated with drug efficiency and clinical performance among VEGFR TK inhibitors

McTigue

Murray

Chen

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

405

282

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Analyses of compounds in clinical development have shown that ligand efficient-molecules with privileged physical properties and low dose are less likely to fail in the various stages of clinical testing, have fewer postapproval withdrawals, and are less likely to receive black box safety warnings. However, detailed side-by-side examination of molecular interactions and properties within single drug classes are lacking. As a class, VEGF receptor tyrosine kinase inhibitors (VEGFR TKIs) have changed the landscape of how cancer is treated, particularly in clear cell renal cell carcinoma, which is molecularly linked to the VEGF signaling axis. Despite the clear role of the molecular target, member molecules of this validated drug class exhibit distinct clinical efficacy and safety profiles in comparable renal cell carcinoma clinical studies. The first head-to-head randomized phase III comparative study between active VEGFR TKIs has confirmed significant differences in clinical performance ) Lancet 378:193-1939. To elucidate how fundamental drug potency-efficiency is achieved and impacts differentiation within the VEGFR TKI class, we determined potencies, time dependence, selectivities, and X-ray structures of the drug-kinase complexes using a VEGFR2 TK construct inclusive of the important juxtamembrane domain. Collectively, the studies elucidate unique drug-kinase interactions that are dependent on distinct juxtamembrane domain conformations, resulting in significant potency and ligand efficiency differences. The identified structural trends are consistent with in vitro measurements, which translate well to clinical performance, underscoring a principle that may be broadly applicable to prospective drug design for optimal in vivo performance.T he VEGF receptor (VEGFR) tyrosine kinases (TKs) are the clinically validated drug target of four structurally diverse TK inhibitors (TKIs) that have been approved in the renal cell carcinoma (RCC) setting (1, 2). The clear cell histology subtype of RCC is molecularly linked to the VEGF signaling axis by virtue of the ∼90% incidence of Von Hippel-Lindau (VHL) dysregulation. With VHL inactivation, hypoxia-inducible factor-α accumulates, leading to overproduction of the angiogenic factor VEGF among others. It is, therefore, generally accepted that on-target VEGFR TK inhibition accounts for the RCC efficacy seen within this class of TKIs. In addition to efficacy in RCC, VEGF signaling inhibition has been linked to side effects, with the most prominent being hypertension, which is consistently seen within the TKI class and the related monoclonal antibody to VEGF, bevacizumab (3).Despite the clear role of VEGF signaling on both hypertension and efficacy in RCC, these on-target pharmacologic effects differ in frequency and degree between approved VEGFR TKI drugs, indicating that the extent of VEGF signal blockade may not be equivalent. Recent reports have analyzed similar RCC clinical studies across leading VEGFR TKIs for comparison purposes (1, 2). Clear distinction in both efficacy...

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Section: Resultssupporting

confidence: 73%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Molecular conformations, interactions, and properties associated with drug efficiency and clinical performance among VEGFR TK inhibitors

McTigue

Murray

Chen

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

405

282

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“…The enzyme phosphorylation (activation) assay for VEGFR2 activity was performed using a R2 construct previously described (McTigue et al, 1999). For this assay, VEGF-R1 and PDGFRβ coupled assay components (2 mM PEP, 300 μM NADH, 5 mM DTT, 15 U/mL PK, 15 U/mL LDH, 200 mM HEPES, pH 7.5) were added to the protein.…”

Section: Biochemical Activity Assays For Human Vegf-r2 Vegf-r1 Pdgfrsmentioning

confidence: 99%

Receptor tyrosine kinase inhibitors AG013764 and AG013711 reduce choroidal neovascularization in rat eye

Wang

Shi

Niesman

et al. 2007

Experimental Eye Research

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Age-related macular degeneration (AMD) is the major cause of blindness for people over 60. In the "wet" form of AMD compounds targeting growth factor signaling pathways such as VEGF have been a major focus for therapeutic interventions. In a previously developed rat model of CNV, we utilized two receptor tyrosine kinase inhibitors (RTKi) to block VEGFR-1, VEGFR-2 and PDGFR signaling following the establishment of CNV. AAV-VEGF 165 was injected into the subretinal space of rats at postnatal days 15-17. Six weeks later, a suspension of RTK inhibitors, AG013764 or AG013711, was injected intraperitoneally (IP, twice daily) or intravitreally (every five days) over a two week period. FITC-dextran whole-mounts of RPE-choroid-sclera were prepared after the animals were sacrificed. CNV area was quantified using Neurolucida to measure the hyperfluorescence on FITC-dextran whole-mounts. Histology and immunohistochemistry were performed as described previously. VEGF expression in control and treated eyes was confirmed by immunohistochemistry and histological sections indicated recovery of retinal morphology and CNV reduction in treated eyes. In the animals IP injected with AG013764 or AG013711 the mean CNV level was reduced by 25 to 33% compared to control, but this effect did not achieve statistical significance. Intravitreal injections of AG013764 or AG013711 reduced the level of CNV by approximately 60% compared to control (p< 0.005 or p< 0.05, respectively). These data show that two RTK inhibitors, AG013764 or AG013711, delivered intravitreally, significantly reduce blood vessel proliferation in this AAV-VEGF 165 model of CNV.

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Overview of Protein Structural and Functional Folds

2004

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This overview provides an illustrated, comprehensive survey of some commonly observed protein-fold families and structural motifs, chosen for their functional significance. It opens with descriptions and definitions of the various elements of protein structure and associated terminology. Following is an introduction into web-based structural bioinformatics that includes surveys of interactive web servers for protein fold or domain annotation, protein-structure databases, protein-structure-classification databases, structural alignments of proteins, and molecular graphics programs available for personal computers. The rest of the overview describes selected families of protein folds in terms of their secondary, tertiary, and quaternary structural arrangements, including ribbon-diagram examples, tables of representative structures with references, and brief explanations pointing out their respective biological and functional significance.

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Crystal structure of the kinase domain of human vascular endothelial growth factor receptor 2: a key enzyme in angiogenesis

Cited by 188 publications

References 47 publications

Molecular conformations, interactions, and properties associated with drug efficiency and clinical performance among VEGFR TK inhibitors

Molecular conformations, interactions, and properties associated with drug efficiency and clinical performance among VEGFR TK inhibitors

Receptor tyrosine kinase inhibitors AG013764 and AG013711 reduce choroidal neovascularization in rat eye

Overview of Protein Structural and Functional Folds

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