In human osteoclast cultures, GW2580 reduced bone resorption together with osteoclast numbers during a three-day exposure. The design does not separate reduced cell number from reduced resorption per surviving cell. [2]
Primary mouse microglia pretreated with 5 µM GW2580 showed increased cell death after hydrogen peroxide challenge, indicating that cellular effects depend on the stress context. [3]
CSF1R enzyme inhibition by GW2580 alone
Biochemical assay · Purchased CSF1R enzyme (SignalChem); ADP-Glo kinase assay [1]
GW2580 inhibited CSF1R kinase activity in a dose-response assay. Fragment combinations and computational predictions are excluded.
Literature-Reported Treatment Conditions
- Treatment concentration
- 0 to 100 µM final compound concentrations described in SI (3-fold serial dilution)
- Exposure duration
- 30 min compound/enzyme preincubation; 40 min kinase reaction
- Method
- Enzymatic ADP formation; ATP reagent described as 200 µM (4X).
- Vehicle
- Stocks and compound dilution series prepared in DMSO; final assay DMSO percentage not explicitly specified.
- Co-treatment
- GW2580 alone (Figure 3A).
- Reported limits
- GW2580 supplier, salt/form and analytical purity not specified in the read scope; exact n and ± definition for this IC50 not specified.
Methods and source locations
Compound reported in the paper: GW2580
Published GW2580 experiment; applicability is to the named compound, not analytical validation of an Adooq batch.
Figure 3A; Results, Determination of Inhibitory Activity and Synergy Using In Vitro Kinase Assays; SI S2
Inhibition of CSF1R-catalyzed ADP formation: ADP-Glo; Hill-equation fit. SI describes two to four independent assays generally; exact Figure 3A n and meaning of ±6.1 are not defined in the read scope. — Figure 3A and caption; SI S2, Kinase assay optimization and IC50 determination
Human osteoclast number and bone resorption
Cell-based study · Human PBMC-derived osteoclast cultures from a healthy female donor; CSF-1 and RANKL differentiation [2]
GW2580 reduced bone resorption and osteoclast numbers in the same culture design; reduced resorption cannot be assigned solely to a per-cell functional effect.
Literature-Reported Treatment Conditions
- Treatment concentration
- 1, 3 and 10 µM GW2580; 0 µM control. Differentiation used CSF-1 and RANKL, each 30 ng/ml.
- Exposure duration
- GW2580 added on day 10; resorption and cell numbers assessed on day 13.
- Method
- Human osteoclast bone-degradation assay; bone surface resorption and immunocytochemical cell counts.
- Vehicle
- DMSO stock diluted to 0.1% DMSO in all wells.
- Co-treatment
- CSF-1 and RANKL present in the osteoclast culture design.
- Reported limits
- One donor described; six cultures do not establish six independent donors.
Methods and source locations
Compound reported in the paper: GW2580
Published GW2580 experiment; applicability is to the named compound, not analytical validation of an Adooq batch.
Table 2 and footnotes; Materials and Methods, Human Osteoclast Bone-Degradation Assay
Bone slice surface resorbed on day 13: Mean ± SEM, six cultures; untreated cultures were statistical controls. Table marks P < 0.01 for each treated group. — Table 2, Bone slice resorbed column and footnotes
23C6-positive cells with >3 nuclei per cm² of bone: Mean ± SEM, six cultures; immunocytochemistry. Table marks P < 0.01 for each treated group. — Table 2, 23C6(+) cells (>3 nuclei) column and footnotes
GW2580 sensitization of primary microglia to peroxide
Cell-based study · Primary microglia from neonatal C57BL/6 mice [3]
GW2580 pretreatment increased peroxide-associated LDH release. This is a stress-dependent result, not evidence of uniform microglial depletion.
Literature-Reported Treatment Conditions
- Treatment concentration
- 5 µM GW2580; hydrogen peroxide 0–100 µM.
- Exposure duration
- 48 h GW2580 pretreatment, followed by 4 h peroxide challenge.
- Method
- Primary microglia culture, imaging and supernatant LDH assay.
- Vehicle
- Medium control; an in vitro GW2580 solvent percentage is not specified in the read methods.
- Co-treatment
- Hydrogen peroxide challenge; paired cultures without GW2580 and zero-peroxide controls.
- Reported limits
- Supplier LC Labs G-5903 is specified in the shared compound/oral-treatment section; culture stock preparation and salt are not specified.
Methods and source locations
Compound reported in the paper: GW2580
Published GW2580 experiment; applicability is to the named compound, not analytical validation of an Adooq batch.
Figure 7C-D and caption; Materials and Methods, Primary Microglia Cultures and Microglia Susceptibility to ROS
Cell death after hydrogen peroxide challenge: Figure 7C-D; three independent experiments, at least duplicate per condition, representing 17 neonatal pups; mean ± SE. — Figure 7C-D and caption; Materials and Methods, Microglia Susceptibility to ROS
