Please cite this article as: Barlaam, B., Cosulich, S., Degorce, S., Ellston, R., Fitzek, M., Green, S., Hancox, U., Lambert-van der Brempt, C., Lohmann, J-J., Maudet, M., Morgentin, R., Plé, P., Ward, L., Warin, N., Discovery of a series of 8-(1-phenylpyrrolidin-2-yl)-6-carboxamide-2-morpholino-4H-chromen-4-one as PI3Kβ/δ inhibitors for the treatment of PTEN-deficient tumours, Bioorganic & Medicinal Chemistry Letters (2017), doi: http:// dx.doi.org/10.1016/j.bmcl.2017.03.027

A novel estrogen receptor down-regulator, 7-hydroxycoumarin (5, SS5020), has been reported with antitumor effects against chemically induced mammary tumors. Here, we report on our own investigation of 7-hydroxycoumarins as potential selective estrogen receptor down-regulators, which led us to the discovery of potent down-regulating antagonists, such as 33. Subsequent optimization and removal of the 7-hydroxy group led to coumarin 59, which had increased potency and improved rat bioavailability relative to SS5020.

A novel estrogen receptor down-regulator, 7-hydroxycoumarin (5, SS5020), has been reported with antitumor effects against chemically induced mammary tumors. Here, we report on our own investigation of 7-hydroxycoumarins as potential selective estrogen receptor down-regulators, which led us to the discovery of potent down-regulating antagonists, such as 33. Subsequent optimization and removal of the 7-hydroxy group led to coumarin 59, which had increased potency and improved rat bioavailability relative to SS5020.

We have developed two different approaches in parallel to rapidly access 2,4-bis aminoaryl pyridine compounds from a common starting material. The C-4/C-2 approach uses palladium-mediated coupling reactions to sequentially functionalize C-4 and then C-2. An alternative C-2/C-4 route uses a regioselective SNAr reaction to first substitute at C-2 then subsequently at C-4 by a palladium-mediated reaction. Both approaches have been used successfully to provide a range of 2,4-bis-aminoaryl pyridine compounds.

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.

Optimization of our bis-anilino-pyrimidine series of EphB4 kinase inhibitors led to the discovery of compound 12 which incorporates a key m-hydroxymethylene group on the C4 aniline. 12 displays a good kinase selectivity profile, good physical properties and pharmacokinetic parameters, suggesting it is a suitable candidate to investigate the therapeutic potential of EphB4 kinase inhibitors.

The synthesis of methyl 2-(5-hydroxy-3-methoxypyridin-2-yl)acetate and alkyl 2-(5-hydroxypyrimidin-2-yl)acetate is described. Methodology for an efficient access to 5-hydroxy-pyridin- and pyrimidin-2-yl acetate cores has been developed. Based on the difference in halogen reactivity, 5-bromo-2-chloropyridine and its pyrimidine analogue were functionalized judiciously by SNAr and palladium-catalyzed reactions. The outlined strategy provides a high-yielding route suitable for large-scale synthesis of these compounds as well as paves the way for a potential rapid access to other heterocyclic analogues.

AZD0530, an orally available Src inhibitor, demonstrated potent antimigratory and anti-invasive effects in vitro, and inhibited metastasis in a murine model of bladder cancer. Antiproliferative activity of AZD0530 in vitro varied between cell lines (IC(50) 0.2 ->10μM). AZD0530 inhibited tumor growth in 4/10 xenograft models tested and dynamically inhibited in vivo phosphorylation of Src substrates paxillin and FAK in both growth-inhibition-resistant and -sensitive xenografts. The activity of AZD0530 in NBT-II bladder cancer cells in vitro was consistent with inhibition of cell migration and stabilization of cell-cell adhesion. These data suggest a dominant anti-invasive pharmacology for AZD0530 that may limit tumor progression in a range of cancers. AZD0530 is currently in Phase II clinical trials.

This paper describes the different strategies devised and applied to overcome the selectivity issues in the syntheses of 6,7-disubstituted-1H-quinolin-4-one, 7-substituted-1H-1,6-naphthyridin-4-one and 6-substituted-1H-1,7-naphthyridin-4-one derivatives. They allowed us to improve the overall yields and the scaling-up feasibility. Several examples illustrate these strategies with their advantages and drawbacks.

Src family kinases (SFKs) are nonreceptor tyrosine kinases that are reported to be critical for cancer progression. We report here a novel subseries of C-5-substituted anilinoquinazolines that display high affinity and specificity for the tyrosine kinase domain of the c-Src and Abl enzymes. These compounds exhibit high selectivity for SFKs over a panel of recombinant protein kinases, excellent pharmacokinetics, and in vivo activity following oral dosing. N-(5-Chloro-1,3-benzodioxol-4-yl)-7-[2-(4-methylpiperazin-1-yl)ethoxy]-5-(tetrahydro-2H-pyran-4-yloxy)quinazolin-4-amine (AZD0530) inhibits c-Src and Abl enzymes at low nanomolar concentrations and is highly selective over a range of kinases. AZD0530 displays excellent pharmacokinetic parameters in animal preclinically and in man (t(1/2) = 40 h). AZD0530 is a potent inhibitor of tumor growth in a c-Src-transfected 3T3-fibroblast xenograft model in vivo and led to a significant increase in survival in a highly aggressive, orthotopic model of human pancreatic cancer when dosed orally once daily. AZD0530 is currently undergoing clinical evaluation in man.

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.