Brilanestrant (GDC-0810)

Catalog No.S7855 Batch:S785501

Technical Data

Formula	C₂₆H₂₀ClFN₂O₂
Molecular Weight	446.90	CAS No.	1365888-06-7
Solubility (25°C)*	In vitro	DMSO	89 mg/mL (199.14 mM)
		Ethanol	89 mg/mL (199.14 mM)
		Water	Insoluble
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description

Brilanestrant (GDC-0810, ARN-810） is a potent ER-α binder (ER-α, IC50 = 6.1 nM; ER-β, IC50 = 8.8 nM), a full transcriptional antagonist with no agonism and displays good potency and efficacy in ER-α degradation (EC50 = 0.7 nM) and MCF-7 breast cancer cell viability (IC50 = 2.5 nM) assays with good selectivity over other nuclear hormone receptors.

Targets

ERα ^[1] (Cell-free)	ERβ ^[1] (Cell-free)
6.1 nM	8.8 nM

In vitro

In cell-free radio-ligand competitive binding assays, GDC-0810 binds both ERα and ERβ with low nanomolar affinity^[2]. GDC-0810 has little to no inhibition against CYP1A2, CYP2D6, or CYP3A4 (IC50 > 20 μM), modest inhibitory effect on CYP2C9 and CYP2C19 (IC50 = 2.2 and 3.3 μM respectively), and potent inhibition of CYP2C8 (IC50 of <0.1 μM). Selectivity of GDC-0810 over other nuclear hormone receptors is also found to be good. In transcriptional reporter assays for the mineralocorticoid (MR), progesterone-A (PR-A), progesterone (PR-B), and glucocorticoid (GR) receptors, GDC-0810 has minimal activity (IC50 > 1 μM). While in binding assays, GDC-0810 displays little activity toward the androgen receptor (AR; IC50 > 4 μM) and GR (IC50 = 0.99 μM)^[1]. GDC-0810-mediated ERα depletion is dependent on the 26S proteasome. GDC-0810 antagonizes ERα ligand binding domain mutants in vitro and in vivo. In cell-free E2 competitive binding assays that was used to determine the binding of GDC-0810 to ER.WT, ER.Y537S and ER.D538G ligand binding domains, GDC-0810 retains its ability to potently displace E2 from the ligand binding domain, albeit with a slightly increased IC50 (WT: 2.6 nM vs. ER.Y537S: 5.5 nM and ER.D538G: 5.4 nM). GDC-0810 can compete the PGC1α co-activator peptide off the mutated ligand binding domain, implying that GDC-0810 is capable of driving an 'active' to 'inactive' conformational shift of mutant ER, though with a ~five-seven fold reduction in biochemical potency compared to wild-type ER^[2].

In vivo

The pharmacokinetic profile of GDC-0810 shows it is a low clearance molecule across species, with good bioavailability (40−60%). As would be expected for a lipophilic carboxylic acid, the compound is highly bound to plasma proteins (>99.5% across species) and has a low to moderate volume of distribution (Vss = 0.2−2.0 L/kg across species). GDC-0810 exhibits good bioavailability across species and displays robust activity in tamoxifen-sensitive and tamoxifen-resistant xenograft models of breast cancer^[1]. GDC-0810 displays mild estrogenic activity in uterine models in vitro and in vivo^[2].

Protocol (from reference)

Cell Assay:^{^[1]}	Cell lines MCF-7 cells Concentrations -- Incubation Time 4 h Method MCF-7 cells are trypsinized and washed twice in phenol red free RPMI containing 5% charcoal dextran stripped FBS with 20 mM HEPES and NEAA and adjusted to a concentration of 200000 cells per mL with the same medium. Next, 16 μL of the cell suspension (3200 cells) is added to each well of a poly-D-lysine coated 384-well plate, and the cells are incubated at 37℃ over 4 days to allow the cells to adhere and grow. On day 4, a 10-point, serial 1:5 dilution of each compound is added to the cells in 16 μL at a final concentration ranging from 10⁻⁵ M to 5.12 × 10⁻¹² M or 10⁻⁶ M to 5.12 × 10⁻¹³ M for fulvestrant. At 4 h post compound addition, the cells are fixed by adding 16 μL of 30% formalin to the 32 μL of cells and compound (10% formalin final concentration) for 20 min. Cells are then washed twice with PBS Tween 0.1% and then permeabilized in PBS 0.1% Triton (50 μL/well) for additional 15 min. The PBS 0.1% triton is decanted, and the cells are washed: LI-COR blocking buffer (50 μL/well) was added, the plate is spun at 3000 rpm, and then the blocking buffer is decanted. Additional LI-COR blocking buffer (50 μL/well) is added, and the cells are incubated overnight at 4℃. The blocking buffer is decanted, and the cells are incubated overnight at 4℃ with SP1 anti-ER rabbit monoclonal antibody diluted 1:1000 in LI-COR blocking buffer/0.1% Tween-20. Wells, which are treated with blocking buffer with Tween but no antibody, are used as a background control. Wells are washed twice with PBS Tween 0.1% to remove free SP1 antibodies, and the cells are incubated at room temp for 60−90 min in LI-COR goat antirabbit IRDyeTM 800CW (1:1000) and DRAQ5 DNA dye diluted in LI-COR blocking buffer containing 0.1% Tween-20 and 0.01% SDS. Cells are then washed with 0.1%Tween-20/PBS three times. Plates are scanned on a LI-COR Odyssey infrared imaging system.
Animal Study:^{^[1]}	Animal Models a tamoxifen-sensitive MCF-7 xenograft model Dosages 30 and 100 mg/kg Administration p.o.

Cell Assay:^{^[1]}

Cell lines
MCF-7 cells
Concentrations
--
Incubation Time
4 h
Method
MCF-7 cells are trypsinized and washed twice in phenol red free RPMI containing 5% charcoal dextran stripped FBS with 20 mM HEPES and NEAA and adjusted to a concentration of 200000 cells per mL with the same medium. Next, 16 μL of the cell suspension (3200 cells) is added to each well of a poly-D-lysine coated 384-well plate, and the cells are incubated at 37℃ over 4 days to allow the cells to adhere and grow. On day 4, a 10-point, serial 1:5 dilution of each compound is added to the cells in 16 μL at a final concentration ranging from 10⁻⁵ M to 5.12 × 10⁻¹² M or 10⁻⁶ M to 5.12 × 10⁻¹³ M for fulvestrant. At 4 h post compound addition, the cells are fixed by adding 16 μL of 30% formalin to the 32 μL of cells and compound (10% formalin final concentration) for 20 min. Cells are then washed twice with PBS Tween 0.1% and then permeabilized in PBS 0.1% Triton (50 μL/well) for additional 15 min. The PBS 0.1% triton is decanted, and the cells are washed: LI-COR blocking buffer (50 μL/well) was added, the plate is spun at 3000 rpm, and then the blocking buffer is decanted. Additional LI-COR blocking buffer (50 μL/well) is added, and the cells are incubated overnight at 4℃. The blocking buffer is decanted, and the cells are incubated overnight at 4℃ with SP1 anti-ER rabbit monoclonal antibody diluted 1:1000 in LI-COR blocking buffer/0.1% Tween-20. Wells, which are treated with blocking buffer with Tween but no antibody, are used as a background control. Wells are washed twice with PBS Tween 0.1% to remove free SP1 antibodies, and the cells are incubated at room temp for 60−90 min in LI-COR goat antirabbit IRDyeTM 800CW (1:1000) and DRAQ5 DNA dye diluted in LI-COR blocking buffer containing 0.1% Tween-20 and 0.01% SDS. Cells are then washed with 0.1%Tween-20/PBS three times. Plates are scanned on a LI-COR Odyssey infrared imaging system.

Animal Study:^{^[1]}

Animal Models
a tamoxifen-sensitive MCF-7 xenograft model
Dosages
30 and 100 mg/kg
Administration
p.o.

References

Selleck's Brilanestrant (GDC-0810) has been cited by 2 publications

G1T48, an oral selective estrogen receptor degrader, and the CDK4/6 inhibitor lerociclib inhibit tumor growth in animal models of endocrine-resistant breast cancer. [ Breast Cancer Res Treat, 2020, 10.1007/s10549-020-05575-9]	PubMed: 32130619
Acquired HER2 mutations in ER+ metastatic breast cancer confer resistance to estrogen receptor-directed therapies. [ Nat Genet, 2019, 51(2):207-216]	PubMed: 30531871

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