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3 protocols using azd9496

1

Comparative Evaluation of Selective ER Modulators

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Fulvestrant (CAS No: 129453–61-8, > 99% purity) was purchased from MedChem Express. Estradiol (E2) (E8875), lasofoxifene (SML1026), 4-hydroxytamoxifen (H7904), and tamoxifen (T5648) were purchased from Sigma. Raloxifene (2280) was purchased from Tocris. GDC-0810 (S7855), bazedoxifene (S2128), and AZD9496 (S8372) were purchased from Selleckchem. GW5638 (5638), GW7604 (7604), and RU 58,668 (RU) were provided by Donald McDonnell (Duke University). G1T48 was provided by G1 Therapeutics, Inc., as analytical grade compound.
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2

Synthesis and Procurement of Diverse Compounds

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SCR-6139, SCR-6515, and SCR-6852 were synthesized as described in (WO2021228210), GDC-9545 was synthesized described in (WO2019245974) (Compound A). AZD9833 was made as described in WO2018077630 (example 17), AZD9496 was purchased from Selleck (#S8372). ARV-471 was synthesized as described in WO2022132652 (compound 1c). Fulvestrant, 4-OH-tamoxifen, Palbociclib, Alpelisib and Elacestrant (RAD1901) were purchased from MCE.
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3

Breast Cancer Cell Wound Healing Assay

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Breast cancer cells were plated onto 96-well cell culture clusters (Costar) and grown to confluence, and then serum-starved for 24 h. Herceptin (Genentech, South San Francisco, CA), AZD9496 (AZD; Selleck, Houston, TX, USA) or megestrol acetate (MA; Selleck) were used 60 min before the scratch was made. The monolayer cells were scratched manually with a plastic pipette tip, and after 2 washes with PBS, the wounded cellular monolayer was allowed to heal for 6 h in DMEM containing indicated dose of kaempferol (Sigma, St Louis, MO, USA). Photographs of central wound edges per condition were taken at time 0 h and 6 h using PowerShot G10 camera (Canon, Tokyo, Japan).
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