In an hMSC-TERT osteoblast differentiation model, 3 µM BMS-833923 reduced ALP activity and mineralized matrix formation versus DMSO controls. The day-10 alamarBlue viability readout was not significantly changed. These findings concern differentiation in an immortalized cell model. [2]
Inverse agonism at wild-type human Smoothened
Biochemical assay · CHO-K1 cell membranes expressing recombinant wild-type human SMO [1]
BMS-833923 reduced constitutive SMO-mediated G-protein activity. Potency in this membrane assay was lower than in downstream cellular readouts; the endpoint is functional inverse agonism, not a direct binding-affinity measurement.
Literature-Reported Treatment Conditions
- Treatment concentration
- Concentration-response experiment; exact tested series not enumerated in the methods.
- Exposure duration
- 30 min ligand preincubation, followed by 30 min [35S]GTPγS incubation.
- Method
- Membrane [35S]GTPγS incorporation assay. Basal incorporation defined as 0%; cyclopamine (10 µM) defined full inverse agonism (−100%).
- Vehicle
- SI: 10^-2 M DMSO stocks, further diluted in assay buffer; final DMSO fraction not specified.
- Reported limits
- Exact concentration series, final solvent fraction, and exact independent n for the Table 1 BMS estimates are not specified in the read locations.
Methods and source locations
Compound reported in the paper: BMS-833923; Compound label in the paper: BMS-833923
Compound-level research evidence; tested supplier material does not validate an Adooq lot.
Table 1, BMS-833923 row, SMO wt columns; Figure 1; Materials and Methods: [35S]GTPγS incorporation; SI Drugs
Inhibition of constitutive wild-type human SMO-mediated [35S]GTPγS incorporation: Nonlinear regression of concentration-response data. — Table 1, BMS-833923 row, SMO wt pIC50 column; Materials and Methods: Data analysis
Maximal inhibition of constitutive wild-type human SMO-mediated G-protein activity: Functional efficacy relative to basal incorporation and 10 µM cyclopamine. — Table 1, BMS-833923 row, SMO wt Emax column; membrane assay methods
Suppression of SHH-induced GLI1 expression in DAOY cells
Cell-based study · Human medulloblastoma-derived DAOY cells stimulated with SHH (50 nM) [1]
BMS-833923 inhibited SHH-induced GLI1 mRNA expression. Its cellular potency should be interpreted within this assay rather than substituted for membrane-assay potency.
Literature-Reported Treatment Conditions
- Treatment concentration
- BMS-833923 concentration response; SHH fixed at 50 nM.
- Exposure duration
- 16 h serum starvation before a 24 h incubation with SHH and compound.
- Method
- qRT-PCR; GLI1 normalized to GAPDH, RPLP0 and YWHAZ. SHH 50 nM stimulation defined as 100%.
- Vehicle
- SI reports DMSO stocks; final DMSO fraction not specified.
- Co-treatment
- SHH stimulation; no second therapeutic inhibitor.
- Reported limits
- Exact BMS concentration series, final solvent fraction and exact independent n for this Table 1 estimate are not specified in the read locations.
Methods and source locations
Compound reported in the paper: BMS-833923; Compound label in the paper: BMS-833923
Compound-level research evidence; tested supplier material does not validate an Adooq lot.
Table 1, BMS-833923 row, DAOY column; Figure 1; Materials and Methods: SHH-induced GLI1 mRNA expression and Data analysis
Inhibition of SHH-induced GLI1 mRNA: qRT-PCR and nonlinear concentration-response fitting. — Table 1, BMS-833923 row, DAOY GLI1 column; Materials and Methods
Reduced osteoblast differentiation with preserved viability readout
Cell-based study · Telomerase-immortalized human bone marrow skeletal mesenchymal stem cells (hMSC-TERT) under osteoblast induction [2]
At 3 µM, BMS-833923 reduced ALP staining/activity at day 10 and mineralized matrix staining at day 21 versus DMSO controls. Day-10 alamarBlue viability did not differ significantly. This is a differentiation model, not primary-donor validation or an IC50 experiment.
Literature-Reported Treatment Conditions
- Treatment concentration
- 3 µM
- Exposure duration
- Throughout osteoblast differentiation; ALP and viability measured at day 10; Alizarin red at day 21.
- Method
- ALP staining and colorimetric activity normalized to cell number; alamarBlue viability; Alizarin Red S matrix staining.
- Vehicle
- DMSO vehicle in osteoblast induction medium; final fraction not specified.
- Co-treatment
- Osteoblast induction medium; no second therapeutic inhibitor.
- Reported limits
- Exact assay well-to-independent-experiment allocation is not given; Figure 1 n = 16 is not assigned donor or biological-replicate status. Exact solvent fraction not specified.
Methods and source locations
Compound reported in the paper: BMS-833923; Compound label in the paper: BMS-833923
Compound-level research evidence; tested supplier material does not validate an Adooq lot.
Sections 2.1–2.8 and 3.1; Figures 1a–c and 2a–b (excluding 2c)
Osteoblast differentiation at day 10: ALP staining and activity normalized to cell number. — Figures 1a–b; sections 2.4–2.5, 3.1
Day-10 viability readout: alamarBlue fluorescence assay. — Figure 1c; sections 2.3, 3.1
Mineralized matrix formation at day 21: Alizarin Red S staining. — Figure 2a; sections 2.6, 3.1
