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Capivasertib

Manufactured by AstraZeneca
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Capivasertib is a laboratory product developed by AstraZeneca. It is an active pharmaceutical ingredient that functions as a protein kinase inhibitor. The core purpose of Capivasertib is for use in research and development applications.

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Lab products found in correlation

Wortmannin, by Selleck Chemicals (1 mentions) Arq 092, by Selleck Chemicals (1 mentions) Mk 2206, by Selleck Chemicals (1 mentions) Celltiter glo assay, by Promega (1 mentions) Palbociclib, by AstraZeneca (1 mentions) Fulvestrant, by AstraZeneca (1 mentions) Afatinib, by Adooq (1 mentions) Caspase glo 3 7 reagent, by Promega (1 mentions) Mk 2206, by Adooq (1 mentions) Azd5363, by AstraZeneca (1 mentions) Adavosertib, by AstraZeneca (1 mentions)

6 protocols using capivasertib

1

Preparation of AstraZeneca Inhibitors

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Selumetinib, AZD8186, and capivasertib (Astra Zeneca Spain), were prepared in dimethyl sulfoxide (DMSO) at 50 mM and stored at room temperature (RT) (selumetinib) or 4°C (AZD8186 and capivasertib) protected from light.
Ruiz de Porras V., Bernat-Peguera A., Alcon C., Laguia F., Fernández-Saorin M., Jiménez N., Senan-Salinas A., Solé-Blanch C., Feu A., Marín-Aguilera M., Pardo J.C., Ochoa-de-Olza M., Montero J., Mellado B, & Font A. (2024). Dual inhibition of MEK and PI3Kβ/δ–a potential therapeutic strategy in PTEN-wild-type docetaxel-resistant metastatic prostate cancer. Frontiers in Pharmacology, 15, 1331648.
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2

AKT Inhibitor Evaluation in Mutant Cell Lines

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Capivasertib was provided by AstraZeneca. MK-2206 and ARQ-092 were purchased from Selleck Chemicals. Compounds were dissolved in DMSO to yield a 10 mM stock, and diluted in assay medium to achieve the desired concentrations. MCF10a cells from stable lines expressing WT or mutant AKT were seeded in 96-well plates and treated with a range of drug concentrations in assay media, and cell viability was assessed 72 h post treatment using the Cell Titer Glo assay (Promega). Viability data were analyzed and IC50 values derived using GraphPad Prism. Wortmannin was purchased from Selleck Chemicals and BYL-719 was kindly provided by the N. Rosen laboratory. MCF10a cells stably expressing AKT1 or AKT2 mutations were seeded in 6-well plates, treated with different concentrations of Wortmannin or BYL-719 in complete growth media, and cell lysates were harvested 2 h post treatment.
Shrestha Bhattarai T., Shamu T., Gorelick A.N., Chang M.T., Chakravarty D., Gavrila E.I., Donoghue M.T., Gao J., Patel S., Gao S.P., Reynolds M.H., Phillips S.M., Soumerai T., Abida W., Hyman D.M., Schram A.M., Solit D.B., Smyth L.M, & Taylor B.S. (2022). AKT mutant allele-specific activation dictates pharmacologic sensitivities. Nature Communications, 13, 2111.
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3

Capivasertib: Bioavailable AKT Inhibitor

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The AKT inhibitor, capivasertib (AZD5363, AstraZeneca, UK), was used for in vitro studies. This compound is a novel synthetic AKT inhibitor that is orally bioavailable and was previously used in clinical trials [19 (link),20 (link)]. For in vitro studies, capivasertib was dissolved in dimethyl sulfoxide (DMSO) at 10 mmol/L stock solution and stored at −20 °C.
Thomas A., Reetz S., Stenzel P., Tagscherer K., Roth W., Schindeldecker M., Michaelis M., Rothweiler F., Cinatl J J.r., Cinatl J., Dotzauer R., Vakhrusheva O., Albersen M., Macher-Goeppinger S., Haferkamp A., Juengel E., Neisius A, & Tsaur I. (2021). Assessment of PI3K/mTOR/AKT Pathway Elements to Serve as Biomarkers and Therapeutic Targets in Penile Cancer. Cancers, 13(10), 2323.
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4

Generating Palbociclib-Resistant Cell Lines

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All cell lines were authenticated using DNA fingerprinting short-tandem repeat (STR) assays. To generate resistant cell populations, parental cell lines were exposed to escalating concentrations of palbociclib over 4–6 months. T47D cells were cultured up to a final dose of 3 μM, which generated two PalboR cell pools (named T47D RB− & T47D CDK6H). MCF7 PalboR cell pools (named MCF7 RB− & MCF7 PacqR) were previously generated and named MCF7 PC8 & MCF7 PC636 (link), respectively, by continuous culture up to a final dose of 1 μM. Capivasertib, palbociclib and fulvestrant (AstraZeneca) were dissolved in dimethyl sulfoxide (DMSO) at a concentration of 10 mmol/L.
Hopcroft L., Wigmore E.M., Williamson S.C., Ros S., Eberlein C., Moss J.I., Urosevic J., Carnevalli L.S., Talbot S., Bradshaw L., Blaker C., Gunda S., Owenson V., Hoffmann S., Sutton D., Jones S., Goodwin R.J., Willis B.S., Rooney C., de Bruin E.C, & Barry S.T. (2023). Combining the AKT inhibitor capivasertib and SERD fulvestrant is effective in palbociclib-resistant ER+ breast cancer preclinical models. NPJ Breast Cancer, 9, 64.
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5

MUC4 Expression Vector Construction

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The pCDH-GFP-MUC4 expression vector was generated by inserting full-length MUC4 beta (NM_138297) into the pCDH-CMV-MCS-EF1α-copGFP vector. GEM, MK-2206 (AKT inhibitor), and afatinib were purchased from AdooQ BioScience (Irvine, CA, US). Caspase-Glo® 3/7 Reagent (G8090) was purchased from Promega Corporation (Wisconsin, United States). Capivasertib (AZD5363, AKT inhibitor) was provided by AstraZeneca plc. (Cambridge, UK).
Pan Y.R., Wu C.E., Jung S.M., Huang S.C., Lin S.H., Chou W.C., Chang Y.C., Chen M.H., Hung T.H., Yu A.L., Huang W.K, & Yeh C.N. (2023). Mucin 4 Confers Gemcitabine Resistance and an Unfavorable Prognosis in Patients with Cholangiocarcinoma via AKT Activation. International Journal of Biological Sciences, 19(9), 2772-2786.
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6

Melanoma Cell Lines and Inhibitors

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The human melanoma cell lines 1205 Lu, WM164, SK-MEL-28, 451 Lu, and murine melanoma line B16F10 were provided by Dr. Meenhard Herlyn (Wistar Institute, Philadelphia, Pennsylvania). The human melanoma cell line UACC 903 was provided by Dr. Mark Nelson (University of Arizona, Tucson, Arizona). The NK-92 cell line was procured from ATCC (Manassas, Virginia). Mouse melanoma cell lines M1 (Mel114433), M3 (HCmel1274), and M4 (B2905) were provided by Dr. Glenn Merlino, National Cancer Institute (NCI, Bethesda, Maryland). M1 tumors were induced by UV radiation in BrafCA/+; Ptenfox/+; Cdkn2afox/+; Tyr-CreERT2-tg mice whereas M4 tumors were induced by UV radiation in Hgftg mice [37 (link)]. Cell lines were maintained in a 37°C humidified 5% CO2 incubator and periodically monitored for phenotypic and genotypic characteristics, as well as tumorigenic potential, to validate and confirm cell line identity. Cell lines were authenticated and tested for mycoplasma contamination periodically and were used within 15 passages after authentication. Clinical-grade pharmacological inhibitors of AKT (AZD5363/capivasertib) and WEE1 (MK1775/adavosertib) were provided by AstraZeneca (Cambridge, United Kingdom). AZD5363 was discovered by AstraZeneca subsequent to a collaboration with Astex Therapeutics (and its collaboration with the Institute of Cancer Research and Cancer Research Technology Limited).
Dinavahi S.S., Chen Y.C., Punnath K., Berg A., Herlyn M., Foroutan M., Huntington N.D, & Robertson G.P. (2022). Targeting WEE1/AKT restores p53-dependent NK cell activation to induce immune checkpoint blockade responses in ‘cold’ melanoma. Cancer immunology research, 10(6), 757-769.
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