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Research Area
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Description
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Cancer |
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Biological Activity
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Description
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Saracatinib (AZD0530) is a Src inhibitor for c-Src with IC50 of 2.7 nM. |
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Targets
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c-Src |
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IC50 |
2.7 nM [1] |
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In Vitro
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Saracatinib also potently inhibits other Src tyrosine kinase family members including c-Yes, Fyn, Lyn, Blk, Fgr, and Lck with IC50 from 4-10 nM. Saracatinib sensitively inhibits Src Y530F NIH 3T3 with IC50 of 80 nM. Saracatinib significantly impairs the invasion of HT1080 cells through a 3-dimensional collagen matrix and completely inhibits EGF-induced cell scattering in NBT-II bladder cancer cells. [1] Saracatinib potent inhibits Src activation in DU145 and PC3 cells, which through inhibition of Y419 phosphorylation. Saracatinib inhibits the growth of prostate cancer including PC3, DU145, CWR22Rv1 and LNCaP, while Saracatinib shows low activity in LAPC-4, PZ-HPV7 and RWPE-1 cells. Saracatinib induces cell cycle arrest at G1/S but not caspase 3 cleavages. Saracatinib also significantly inhibits DU145 and PC3 migration in the Boyden chamber. [2] Saracatinib gives a potent and sustained blockage of AKT and enhancs the sensitivity to irradiation in A549 and Calu-6 cells. [3] Saracatinib inhibits osteoclast in activity, resorption and formation. Saracatinib also reversibly prevents osteoclast precursor migration. [4] |
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In Vivo
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Saracatinib shows great tumor growth inhibiton in Src3T3 allografts and a moderate growth delay in Calu-6, MDA-MB-231, AsPc-1 and BT474C xenografts. [1] Saracatinib shows great antitumor activity in orthotopic DU145 xenograft mice at a dose of 25mg/kg (orally administered, daily). [2] |
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Clinical Trials
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Saracatinib is currently in Phase II clinical trial in recurrent osteosarcoma localized to the lung. |
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Features
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Saracatinib is the first Src inhibitor to show inhibition of the Src pathway in human tumor tissue. |
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Combination Therapy
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Description
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AZD0530 at concentration of 1 μM combined with Imatinib at concentration of 0.1 μM show an additive inhibitory effect on the proliferation of CML BV173 cells. [5] AZD0530 at concentration of 1 μM in combination with Tamoxifen at concentration ranging from 1 nM to 10 μM show improved growth inhibitory effects in both MCF7 and T47D cells, with decreased Ki67 expression. AZD0530 combined with Tamoxifen result in a reduction of Src activity together with inhibition of focal adhesion kinase phosphorylation and a complete abrogation of their in vitro invasive behaviour in both MCF7 and T47D cells. [6] AZD0530 at concentration of 1 μM in combination with Anastrozole at concentration of 100 μM increase p27 and cause greater G(1) cell cycle arrest in MCF-7Arom5 cells. AZD0530 administrated at dose of 50 mg/kg by oral gavage in combination with Anastrozole subcutaneously injected at dose of 0.25 mg/kg or 5 μg/animal reduce drug resistance and show greater antitumor efficacy with greater Src and EGFR inhibition and a greater increase in p27 and reduction of Ki-67 in MCF-7Arom5 tumor xenografts mice. [7] |
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Protocol
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Kinase Assay
[1]
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| Kinase Assay |
IC50 of tyrosine kinase activity is measured by an enzyme-linked immunosorbent assay (ELISA) with recombinant catalytic domains of a panel of receptor and non-receptor tyrosine kinases (in some cases only part of the catalytic domain is used). Saracatinib dose ranges from 0.001–10 mM. Speci?city assays against a panel of serine/threonine kinases are performed using a ?lter capture assay with 32P. Briefly, multidrop 384 plates containing 0.5 μL Saracatinib or controls (DMSO) alone or pH 3.0 buffer controls) are incubated with 15 μL of enzyme plus peptide/protein substrate for 5 min before the reaction is initiated by the addition of 10 μL of 20 mM Mg-ATP. For all enzymes the ?nal concentration is approximated to the Michaelis constant (Km). Assays are carried out for 30min at room temperature before termination by the addition of 5 μL orthophosphoric acid. After mixing, the well contents are harvested onto a P81 Uni?lter plate, using orthophosphoric acid as the wash buffer. Then IC50 is calculated. |
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Cell Assay
[1]
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| Cell Lines |
PC3, DU145, CWR22Rv1, LNCaP, LAPC-4, PZ-HPV7 and RWPE-1 cells |
| Concentrations |
62.5 nM - 16 mM |
| Incubation Time |
1, 3 and 5 days |
| Methods |
Cells are seeded at a density of 2×103 in 96-well plates and incubated overnight. Then Saracatinib (62.5 nM–16 mM) is added to the cells. After 1, 3 and 5 days, culture medium is removed followed by addition of 0.2 mL DMSO per well and continuous shaking of plates at 200 rotations per minute for 15min. Then IC50 is measured by MTT method. |
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Animal Study
[1]
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| Animal Models |
CB17 mice are implanted with DU145 cells. |
| Formulation |
Dissolved in 0.5% hydroxypropyl methylcellulose, 0.1% Tween 80 |
| Doses |
25 mg/kg |
| Administration |
Orally administered daily |
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References |
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[1] Chang YM, Oncogene, 2008, 27(49), 6365-6375.
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[2] Green TP, et al. Mol Oncol, 2009, 3(3), 248-261.
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[3] Purnell PR, et al. J Thorac Oncol, 2009, 4(4), 448-454.
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[4] de Vries TJ, et al. Mol Cancer Res, 2009, 7(4), 476-488.
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[5] Gwanmesia PM, et al. BMC Cancer, 2009, 13;9:53.
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[6] Hiscox S, et al. Breast Cancer Res Treat, 2009, 115(1), 57-67.
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[7] Chen Y, et al. Clin Cancer Res, 2009, 15(10), 3396-3405.
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