Alisertib (MLN8237)

N.º de catálogoS1133 Lote:S113313

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Datos técnicos

Fórmula

C27H20ClFN4O4
Peso molecular 518.92 Número CAS 1028486-01-2
Solubilidad (25°C)* In vitro DMSO 100 mg/mL (192.7 mM)
Water Insoluble
Ethanol Insoluble
In vivo (Agregue los solventes al producto individualmente y en orden.)
Homogeneous suspension
CMC-NA
≥5mg/ml Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution
5%DMSO 30%PEG300 5%Tween80 60%ddH2O

Validado por los laboratorios Selleck. Si necesita ajustes en esta formulación, póngase en contacto con nuestro equipo de ventas para pruebas personalizadas.

 6.000mg/ml (11.56mM) Taking the 1 mL working solution as an example, add 50 μL of 120 mg/ml clarified DMSO stock solution to 300 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 600 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results. 
* <1 mg/ml significa ligeramente soluble o insoluble.
* Tenga en cuenta que Selleck prueba la solubilidad de todos los compuestos internamente, y la solubilidad real puede diferir ligeramente de los valores publicados. Esto es normal y se debe a ligeras variaciones entre lotes.
* Envío a temperatura ambiente (Las pruebas de estabilidad demuestran que este producto se puede enviar sin medidas de refrigeración.)

Preparación de soluciones madre

Actividad biológica

Descripción Alisertib (MLN8237) es un inhibidor selectivo de Aurora A con una IC50 de 1,2 nM en un ensayo sin células, y tiene una selectividad >200 veces mayor por Aurora A que por Aurora B. Este compuesto induce la detención del ciclo celular, la apoptosis y la autophagy. Fase 3.
Objetivos
Aurora A
(Cell-free assay)
1.2 nM
In vitro

Alisertib (MLN8237) muestra una selectividad >200 veces mayor por Aurora A que por la Aurora B estructuralmente relacionada con una IC50 de 396,5 nM, y no tiene actividad significativa contra otras 205 quinasas. El tratamiento con este compuesto (0,5 μM) inhibe la fosforilación de Aurora A en células MM1.S y OPM1, sin afectar la fosforilación de la histona H3 mediada por Aurora B. Inhibe significativamente la proliferación celular en líneas celulares de mieloma múltiple (MM) con valores de IC50 de 0,003-1,71 μM, y muestra una actividad antiproliferativa más potente contra células MM primarias y líneas celulares MM en presencia de células del estroma de la médula ósea, así como IL-6 e IGF-1, que contra células MM solas. A 0,5 μM, induce un aumento de 2 a 6 veces en la fase G2/M en células MM primarias y líneas celulares, así como apoptosis y senescencia significativas, lo que implica la regulación al alza de p53, p21 y p27, así como el clivaje de PARP, caspasa 3 y caspasa 9. Además, muestra un fuerte efecto anti-MM sinérgico con Hexadecadrol, así como un efecto aditivo con doxorrubicina y LDP-341. El tratamiento con este compuesto (0,5 μM) causa la inhibición de la formación de colonias de las líneas celulares de adenocarcinoma esofágico FLO-1, OE19 y OE33, e induce un aumento significativo en el porcentaje de células poliploides, y posteriormente un aumento en el porcentaje de células en la fase sub-G1, lo que puede mejorarse aún más en combinación con NSC 119875 (2,5 μM), lo que implica una mayor inducción de TAp73β, PUMA, NOXA, caspasa-3 clivada y PARP clivada en comparación con un tratamiento de agente único.

In vivo

Alisertib (MLN8237) reduce significativamente la carga tumoral con una inhibición del crecimiento tumoral (TGI) del 42% y 80% a 15 mg/kg y 30 mg/kg, respectivamente, y prolonga la supervivencia de los ratones en comparación con el control.

Características Primer inhibidor de Aurora A disponible por vía oral.

Protocolo (de referencia)

Ensayo de quinasa:

[1]
  • Ensayo enzimático de Flashplate radiactivo de Aurora A

    Se realiza un ensayo enzimático de Flashplate radiactivo de Aurora A para determinar la naturaleza y el grado de inhibición mediada por Alisertib (MLN8237) in vitro. La Aurora A recombinante se expresa en células Sf9 y se purifica mediante cromatografía de afinidad GST. El sustrato peptídico para Aurora A se conjuga con biotina (Biotin-GLRRASLG). La quinasa Aurora A (5 nM) se ensaya en 50 mM Hepes (pH 7,5), 10 mM MgCl2, 5 mM DTT, 0,05% Tween 20, 2 μM de sustrato peptídico, 3,3 μCi/mL [γ-33P]ATP a 2 μM, y concentraciones crecientes de este compuesto utilizando Image FlashPlates.
Ensayo celular:

[2]
  • Líneas celulares

    MM1.S, MM.1R, LR5, RPMI 8226, DOX40, OPM1, OPM2, INA6, and U266

  • Concentraciones

    Dissolved in DMSO, final concentrations ~10 μM

  • Tiempo de incubación

    24, 48, and 72 hours

  • Método

    Cells are exposed to various concentrations of Alisertib (MLN8237) for 24, 48, and 72 hours. Cell viability is measured using MTT assay, and cell proliferation is measured using 3[H]-thymidine incorporation. For cell cycle analysis, cells are permeabilized by 70% ethanol at -20 °C, and incubated with 50 μg/mL PI and 20 units/mL RNase-A. DNA content is analyzed by flow cytometry using BDFACS-Canto II and FlowJo software. For the detection of apoptosis and senescence, cells are stained with Resorcinolphthalein isothiocyanate-annexin V and PI. Apoptotic cells are determined by flow cytometric analysis using BDFACS-Canto II and FlowJo software.
Estudio en animales:

[2]
  • Modelos animales

    Severe combined immune-deficient (SCID) mice inoculated subcutaneously with MM1.S cells

  • Dosificaciones

    ~30 mg/kg/day

  • Administración

    Orally

Referencias

  • https://pubmed.ncbi.nlm.nih.gov/22016509/
  • https://pubmed.ncbi.nlm.nih.gov/20382844/
  • https://pubmed.ncbi.nlm.nih.gov/22302096/

Validación de productos por parte del cliente

<p>Alisertib inhibits AURKA and AURKB in a concentration-dependent manner. (a) Alisertib induces G 2 /M delay or genome reduplication. HeLa cells were exposed to buffer or the indicated concentrations of Alisertib. After 24 h, the cells were harvested and analyzed with flow cytometry. The positions of 2N, 4N and 8N DNA contents are indicated. (b) Alisertib delays mitotic exit or induces slippage. HeLa cells stably expressing histone H2B-GFP were exposed to buffer or the indicated concentrations of Alisertib. Individual cells were then tracked for 24 h with time-lapse microscopy. Each horizontal bar represents one cell (n ¼ 50). Key: light gray ¼ interphase; black ¼ mitosis (from DNA condensation to anaphase or mitotic slippage); dark gray ¼ interphase after mitotic slippage; truncated bars ¼ cell death. (c) Different concentrations of Alisertib are involved in delaying mitotic exit and inducing slippage. Live-cell imaging of cells treated with Alisertib was described in panel (b). The duration of mitosis (mean±90% confidence interval) and the percentage of cells that underwent mitotic slippage during the imaging period was quantified. (d) Alisertib promotes apoptosis in a concentration-dependent manner. HeLa cells were incubated with the indicated concentrations of Alisertib for 48 h. The cells were then harvested and analyzed with flow cytometry. (e) Concentration-dependent cytotoxicity of Alisertib. HeLa cells were cultured in the presence of the indicated concentrations of Alisertib for 48 h. The number of live and dead cells was analyzed with trypan blue exclusion assay. (f) Concentration-dependent suppression of long-term survival by Alisertib. HeLa cells were seeded on 60-mm culture plates and grown in the presence of 250 n M or 1 m M of Alisertib. After 24 h, the cells were washed gently and propagated in normal medium for another 10–12 days. Colonies were fixed and stained with crystal violet solution (examples of the plates are shown). Average±s.d. from three independent experiments. (g) Both AURKA and AURKB are inhibited by Alisertib.Mitotic HeLa cells were obtained by exposure to nocodazole for 16 h followed by mechanical shake off. The cells were incubated with the indicated concentrations of Alisertib for 2 h. Lysates were then prepared and activated phospho-AURKAThr288 and AURKBThr232were detected with immunoblotting. The asterisk indicates the position of an AURKB-like protein (the same throughout this study). Uniform loading was confirmed by immunoblotting for actin. In this assay, nocodazole and MG132 (a proteasome inhibitor) were added to prevent the cells from exiting mitosis. Accordingly, the total AURKA and AURKB levels remained constant throughout the experiment. (h) Alisertib prevents activation of AURKA and AURKB. HeLa cells were incubated with the indicated concentrations of Alisertib for 8 h. Nocodazole was then added for another 6 h to trap cells that entered mitosis. Lysates were prepared and analyzed with immunoblotting. Actin analysis was included to assess loading and transfer.</p>

Datos de [ Oncogene , 2014 , 33, 3550-60 ]

Tissue levels of 53BP1, a-tubulin, IkB-a and IL-6 in an Hs294T xenograft treated with MLN8237 or vehicle control were visualized by immunofluorescence co-staining with DAPI. Representative micrographs are shown from triplicate experiments.

Datos de [ EMBO Mol Med , 2013 , 5(1), 149-66 ]

Inhibition of Aurka kinase activity by MLN8237 impairs expression of pluripotency genes in CCE cells as measured by qRT-PCR. All values shown are mean ?SEM for n=3. The level of phosphorylated H3(S10) (p-H3(S10)), an Aurka phosphorylation target site, is decreased in MLN8237-treated samples.

Datos de [ Cell Stem Cell , 2012 , 11, 179-94 ]

<p>Recruitment of clathrin to the mitotic spindle is controlled by phosphorylation of TACC3 by Aurora-A kinase. Representative micrographs of HEK293 cells incubated with 0.3 μM MLN8237 for 40 min. Cells were fixed and stained as indicated.</p>

Datos de [ EMBO J , 2012 , 30, 906-19 ]

Sellecks Alisertib (MLN8237) Ha sido citado por 380 Publicaciones

Centromere protection requires strict mitotic inactivation of the Bloom syndrome helicase complex [ Nat Commun, 2025, 16(1):7832] PubMed: 40846865
Aurora A regulates the material property of spindle poles to orchestrate nuclear organization at mitotic exit [ EMBO J, 2025, 10.1038/s44318-025-00564-4] PubMed: 40940421
Targeted inhibition of Aurora kinase A promotes immune checkpoint inhibition efficacy in human papillomavirus-driven cancers [ J Immunother Cancer, 2025, 13(1)e009316] PubMed: 39773561
An Aurora kinase A-BOD1L1-PP2A B56 axis promotes chromosome segregation fidelity [ Cell Rep, 2025, 44(2):115317] PubMed: 39970043
The AURKA inhibitor alters the immune microenvironment and enhances targeting B7-H3 immunotherapy in glioblastoma [ JCI Insight, 2025, e173700] PubMed: 39928563
CDK1-mediated phosphorylation of LDHA fuels mitosis through LDHB-dependent lactate oxidation [ EMBO Rep, 2025, 10.1038/s44319-025-00573-8] PubMed: 40940446
Cellular senescence as a prognostic marker for predicting breast cancer progression in 2D and 3D organoid models [ Biomed Pharmacother, 2025, 189:118324] PubMed: 40616881
Actionable heterogeneity of hepatocellular carcinoma therapy-induced senescence [ Cancer Immunol Immunother, 2025, 74(7):207] PubMed: 40374812
Aurora B maintains spherical shape of mitotic cells via simultaneously stabilizing myosin II and vimentin [ J Mol Cell Biol, 2025, mjaf023] PubMed: 40795355
O 6-methylguanine DNA methyltransferase (MGMT) expression in U1242 glioblastoma cells enhances in vitro clonogenicity, tumor implantation in vivo, and sensitivity to alisertib-carboplatin combination treatment [ Front Cell Neurosci, 2025, 19:1552015] PubMed: 40336841

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