Supplementary Tables 1-3 from Assessing the Activity of Cediranib, a VEGFR-2/3 Tyrosine Kinase Inhibitor, against VEGFR-1 and Members of the Structurally Related PDGFR Family

Supplementary Figure 1 from Assessing the Activity of Cediranib, a VEGFR-2/3 Tyrosine Kinase Inhibitor, against VEGFR-1 and Members of the Structurally Related PDGFR Family

During our investigation to find suitable conditions to prepare very high molecular weight partially de-acetylated hyaluronic acid (HA), we discovered a powerful new method to cleave amide bonds using hydroxylamine salts at neutral pH with remarkable selectivity over common carbamate protecting groups and other carbonyl functionality including unhindered esters.

Starting from TGX-221, we designed a series of 9-(1-anilinoethyl)-2-morpholino-4-oxo-pyrido[1,2-a]pyrimidine-7-carboxamides as potent and selective PI3Kβ/δ inhibitors. Structure–activity relationships and structure–property relationships around the aniline and the amide substituents are discussed. We identified compounds 17 and 18, which showed profound pharmacodynamic modulation of phosphorylated Akt in the PC3 prostate tumour xenograft, after a single oral dose. Compound 17 also gave significant inhibition of tumour growth in the PC3 prostate tumour xenograft model after chronic oral dosing.

Deregulation of HER family signaling promotes proliferation and tumor cell survival and has been described in many human cancers. Simultaneous, equipotent inhibition of EGFR-, HER2-, and HER3-mediated signaling may be of clinical utility in cancer settings where the selective EGFR or HER2 therapeutic agents are ineffective or only modestly active. We describe the discovery of AZD8931 (2), an equipotent, reversible inhibitor of EGFR-, HER2-, and HER3-mediated signaling and the structure-activity relationships within this series. Docking studies based on a model of the HER2 kinase domain helped rationalize the increased HER2 activity seen with the methyl acetamide side chain present in AZD8931. AZD8931 exhibited good pharmacokinetics in preclinical species and showed superior activity in the LoVo tumor growth efficacy model compared to close analogues. AZD8931 is currently being evaluated in human clinical trials for the treatment of cancer.

A new series of quinoline ether inhibitors, which potently and selectively inhibit PDGFR tyrosine kinases, is described in this Letter. Compounds 23 and 33 are selective, low nanomolar inhibitors of PDGFRα and β, display good pharmacokinetics in rat and dog and are active in vivo at low doses when given orally twice daily. Further evaluation of these compounds is warranted.

A new series of Quinazoline Ether Inhibitor which potently inhibits VEGFR-2 and PDGFR tyrosine kinases is described here. In vitro, pharmacokinetics and in vivo evaluations led to the selection of AZD2932.

Abstract Cediranib is a potent inhibitor of the VEGF receptor (VEGFR)-2 and VEGFR-3 tyrosine kinases. This study assessed the activity of cediranib against the VEGFR-1 tyrosine kinase and the platelet-derived growth factor receptor (PDGFR)-associated kinases c-Kit, PDGFR-α, and PDGFR-β. Cediranib inhibited VEGF-A–stimulated VEGFR-1 activation in AG1-G1-Flt1 cells (IC50 = 1.2 nmol/L). VEGF-A induced greatest phosphorylation of VEGFR-1 at tyrosine residues Y1048 and Y1053; this was reversed by cediranib. Potency against VEGFR-1 was comparable with that previously observed versus VEGFR-2 and VEGFR-3. Cediranib also showed significant activity against wild-type c-Kit in cellular phosphorylation assays (IC50 = 1–3 nmol/L) and in a stem cell factor–induced proliferation assay (IC50 = 13 nmol/L). Furthermore, phosphorylation of wild-type c-Kit in NCI-H526 tumor xenografts was reduced markedly following oral administration of cediranib (≥1.5 mg/kg/d) to tumor-bearing nude mice. The activity of cediranib against PDGFR-β and PDGFR-α was studied in tumor cell lines, vascular smooth muscle cells (VSMC), and a fibroblast line using PDGF-AA and PDGF-BB ligands. Both receptor phosphorylation (IC50 = 12–32 nmol/L) and PDGF-BB–stimulated cellular proliferation (IC50 = 32 nmol/L in human VSMCs; 64 nmol/L in osteosarcoma cells) were inhibited. In vivo, ligand-induced PDGFR-β phosphorylation in murine lung tissue was inhibited by 55% following treatment with cediranib at 6 mg/kg but not at 3 mg/kg or less. In contrast, in C6 rat glial tumor xenografts in mice, ligand-induced phosphorylation of both PDGFR-α and PDGFR-β was reduced by 46% to 61% with 0.75 mg/kg cediranib. Additional selectivity was showed versus Flt-3, CSF-1R, EGFR, FGFR1, and FGFR4. Collectively, these data indicate that cediranib is a potent pan-VEGFR kinase inhibitor with similar activity against c-Kit but is significantly less potent than PDGFR-α and PDGFR-β. Mol Cancer Ther; 10(5); 861–73. ©2011 AACR.

The partnership between rational synthesis design and mass-triggered preparative LCMS is a powerful one, capable of furnishing very large libraries in a selective manner in a very short space of time. Herein, we communicate one example of possibly a perfect marriage between the synthetic chemistry and the subsequent purification method employed, affording a ∼1000-member library supplying 50 mg on average of final compound in less than a month.

Over the last two decades, substituted quinazolines have attracted much interest as kinase inhibitors from the pharmaceutical industry and academic groups. The quinazoline core represents a relatively simple and accessible scaffold that can be substituted via a number of key vectors to give potent and selective inhibitors of a wide range of kinase targets. Here, we will attempt to condense over 20 years of medicinal chemistry library research on this scaffold carried out in AstraZeneca's laboratories and externally into what we regard as the key "savoir-faire".

Synthetic approaches toward the natural product cytostatin, an inhibitor of protein phosphatase 2A possessing cytotoxic and antimetastatic activities, have been investigated. A formal synthesis of cytostatin has been achieved according to a strategy relying on the formation of the C8-C9 bond by a nucleophilic addition of a functionalized organolithium (C1-C8 subunit) to an aldehyde (C9-C13 subunit). (C) 2008 Elsevier Ltd. All rights reserved.

AIM:To evaluate the clinical results of embolisation of symptomatic, incompetent pelvic veins in women presenting with perineal veins.PATIENTS AND METHODS:Twenty-four women presenting with non-saphenous perineal varicose veins and who experienced pelvic vein syndrome were treated. Symptoms was scored on a visual analogue scale assessing dyspareunia, pelvic and lower limb pain. Lower limb varices were investigated by duplex ultrasonography. Pelvic veins were studied by pelvic vein angiography with simultaneous embolisation of incompetent veins. Ovarian and internal iliac veins were systematically embolised when incompetent. Follow-up assessment of symptoms and varices was carried out at 1, 2 and 3 years.RESULTS:All patients presented with perineal veins, 2 with sciatic vein incompetence and 2 with a perforator of the thigh or buttock. Pelvic venous angiography was performed via right femoral access in 87% of the cases and confirmed the presence of incompetent ovarian and internal iliac veins. The mean number of coils used per vein was 6 and all were successfully embolised. No serious complications were encountered. The mean clinical improvement score was 80%, 77%, 80% and 76% at respectively 45 days, 1, 2 and 3-year follow-up.CONCLUSION:In women of reproductive age, non-saphenous varicose veins associated with pelvic venous incompetence (PVI) should undergo pelvic vein investigation. In this clinical series we achieved a satisfactory improvement in symptoms after 3 years following treatment of incompetent pelvic veins.

The structure-activity and structure-property relationships of anilinoquinazoline inhibitors of EGFR were investigated. Strategies to lower volume of distribution and shorten half-life through structure and pKa modulation are discussed.

2537 Normal cell growth, activity and function are coordinated and regulated by a complex network of signaling pathways. Aberrations in these signaling pathways are a hallmark of tumor development. Src kinase, a non-receptor tyrosine kinase, is a signal transduction modulator and is a critical component of many of the key signaling pathways currently thought to be pivotal in carcinogenesis. In contrast to its highly regulated role in normal cells, Src kinase has been shown to be de-regulated and its activity significantly increased in many human tumors. Recent publications have highlighted the critical role of Src kinase in tumor cell migration and invasion. Inhibition of Src kinase has the potential to have a significant impact as a treatment for cancer. AZD0530, a novel anilino quinazoline, is a potent dual-specific inhibitor of cSrc kinase (IC50: 2.7 nM) and the related Abl kinase (IC50: 30 nm). AZD0530 is highly selective for Src and Abl kinases against a large range of tyrosine and serine-threonine kinases (VEGFR2, FGFR, c-Kit, Aur-3 etc. >5 μM). AZD0530 exerts its activity through ATP competitive and reversible inhibition of the target enzyme. Structure-activity analyses that led to the discovery of AZD0530 are presented, focusing in particular on the critical role played by the aniline substitution pattern and the effect of the C5-C7 quinazoline substitution pattern. AZD0530 possesses excellent properties for a small molecule inhibitor of Src kinase that translate into excellent ADME properties in preclinical animal species. AZD0530 is orally available and is suitable for once-daily administration to humans. AZD0530 causes dramatic inhibition of human breast cancer cell (MDA-MB231) migration in vitro. Following oral administration at single daily doses of 6 mg/kg, AZD0530 completely prevents the growth of c-Src-NIH 3T3 xenografts grown in nude rats, and at 25 mg/kg/d significantly increases survival in an orthotopic model of human pancreatic cancer (BxPC-3). AZD0530 is a highly selective, orally available, dual-specific Src/Abl kinase inhibitor. It has the potential for activity in a wide range of tumors. AZD0530 is currently in early clinical development.

A series of 5,7-disubstituted quinazolines, bearing 4-heteroaryl substituents such as 2-pyridinylamine or 2-pyrazinylamine, has been synthetised and evaluated as c-Src kinase inhibitors. Highly potent inhibition, high selectivity and physical properties suitable for oral dosing were achieved within this series: 23d and 42 were identified as sub-0.1muM inhibitors in a c-Src-driven cell proliferation assay and displayed adequate rat pharmacokinetics after oral administration.

4555 AZD2171 is a sub-nanomolar inhibitor of vascular endothelial growth factor (VEGF)-induced KDR (VEGFR-2) signaling in human endothelial cells in vitro . Signaling from this receptor is considered to be the major stimulus of physiological and pathological angiogenesis. AZD2171 may therefore have potential as an inhibitor of tumor neovascularization in a range of malignancies. To demonstrate inhibition of VEGF signaling by AZD2171 in vivo and to examine the consequences of chronic administration, a range of endpoints were studied. Direct inhibition of VEGF signaling in vivo was shown using an acute growth factor hypotension model in anesthetized rats. AZD2171 (250 μg/kg i.v.) completely reversed a hypotensive change induced by intravenous bolus injection of VEGF (32 μg/kg). Chronic inhibition of VEGF signaling has been shown to prevent ossification during long bone growth, by inhibiting vascular invasion of the growth plate. AZD2171 (1.25–5 mg/kg/day) inhibited endochondral ossification significantly, producing a dose-dependent increase in the hypertrophic chondrocyte zone. Morphometric image analysis of femoral and tibial growth plates indicated that AZD2171 (5 mg/kg/day for 28 days) increased the combined growth plate area by more than 400%. This phenotype was fully reversed when followed by an additional period of 28 days without drug treatment. In female rats, significant inhibition of corpora lutea formation was observed in the ovary following AZD2171 treatment (5 mg/kg/day for 28 days): an additional physiological process that is dependent upon angiogenesis. To evaluate antitumor activity in the rat, athymic animals bearing established Calu-6 human lung cancer xenografts (0.23–0.5 cm 3 volume) were treated once daily with AZD2171 (0.1–3 mg/kg p.o.) for 28 days. A dose-dependent inhibition of tumor growth was observed, with statistically significant activity at all doses ( P 90% growth inhibition at 3 mg/kg/day. Calu-6 tumors retrieved from athymic mice following chronic AZD2171 treatment were examined by immunohistochemistry. Tumors from mice treated with 6 mg/kg/day AZD2171 (for 28 days), were found to have significantly reduced CD31 vessel staining and a six-fold increase in tumor cell necrosis, when compared with control tumors. These observations are compatible with inhibiting VEGF-dependent neovascularization and vascular permeability in tumors. Collectively, the preclinical data obtained with AZD2171 are consistent with potent inhibition of VEGF signaling and angiogenesis in vivo . AZD2171 is currently in clinical development as a once daily oral therapy for the treatment of cancer.