At 20 μM TGX-221 plus 50 μM temozolomide for four days, PI3Kβ-high SF295 and U87MG glioblastoma cells showed positive Bliss interactions (45.5% and 26.3% excess over Bliss). PI3Kβ-low LN229/A172 cells lacked appreciable positive interaction; astrocyte viability was not significantly altered in this comparison. The result is specific to these models and this combination. [2]
TGX-221 at 0.5 μM for six hours did not suppress SDF1α-stimulated ERK1/2 phosphorylation in DU145, PC3 or HEK293 cells. This pathway-specific negative result limits generalization of TGX-221 effects across signaling contexts. [3]
Recombinant class IA PI3K lipid-kinase inhibition
Biochemical assay · Recombinant p85/p110 preparations: murine p110α with human p85α, human p110β with human p85α, and commercial p85α/p110δ. [1]
TGX-221 preferentially inhibited p110β in this assay; inhibition of p110δ occurred at a higher concentration. The reported endpoints are enzyme IC50 values, not cellular treatment concentrations.
Literature-Reported Treatment Conditions
- Method
- Radiolabeled lipid-kinase assay using phosphatidylinositol substrate, 100 μM ATP and [γ-33P]ATP; TLC/phosphorimager quantification and nonlinear regression.
- Vehicle
- 1% DMSO
- Reported limits
- Assay incubation duration and full concentration series are not stated in the scoped main assay description; no γ IC50 is provided for TGX-221 in Table 1.
Methods and source locations
Compound reported in the paper: TGX-221
Evidence for the named research compound in the reported model; no claim of Adooq lot equivalence or clinical efficacy.
Table 1 TGX-221 row and footnote; Methods: Production of recombinant PI3K; Synthesis and biochemical characterization of PI3K inhibitors; Figure 1
p110β lipid-kinase inhibition: Table 1: mean ± SD; n≥3 independent experiments across multiple recombinant-protein preparations. — Table 1 TGX-221 row and footnote; Methods: Production of recombinant PI3K; Synthesis and biochemical characterization of PI3K inhibitors; Figure 1
p110α lipid-kinase inhibition: Same assay; preserve the reported greater-than operator. — Table 1 TGX-221 row and footnote; Methods: Production of recombinant PI3K; Synthesis and biochemical characterization of PI3K inhibitors; Figure 1
p110δ lipid-kinase inhibition: Table 1: mean ± SD; n≥3. — Table 1 TGX-221 row and footnote; Methods: Production of recombinant PI3K; Synthesis and biochemical characterization of PI3K inhibitors; Figure 1
TGX-221 did not suppress CXCR4-stimulated ERK1/2 phosphorylation
Cell-based study · DU145 and PC3 human prostate cancer cells and HEK293 cells. [3]
TGX-221 did not visibly suppress SDF1α-induced ERK1/2 phosphorylation under the tested conditions. PI3Kγ and PI3Kδ inhibitor comparators reduced the response. This is a pathway-specific negative result.
Literature-Reported Treatment Conditions
- Treatment concentration
- 0.5 μM TGX-221
- Exposure duration
- 6 h pretreatment; SDF1α stimulation for 5 min
- Method
- Serum-starved cells stimulated with 200 ng/ml SDF1α; phospho-ERK1/2 and total ERK1/2 Western blotting.
- Reported limits
- TGX-221 solvent and final vehicle concentration not specified in the scoped methods/caption. Direct p110β target engagement in this experiment was not measured.
Methods and source locations
Compound reported in the paper: TGX-221
Evidence for the named research compound in the reported model; no claim of Adooq lot equivalence or clinical efficacy.
Figure 4A image/caption; Results sec1.4; Experimental procedures sec3.1, sec3.3, sec3.7, sec3.10
SDF1α-induced ERK1/2 phosphorylation: Figure 4A; basal and stimulated control lanes, total ERK1/2 control; representative blots of at least three experiments. No quantitative TGX-specific effect size or p value reported. — Figure 4A image/caption; Results sec1.4; Experimental procedures sec3.1, sec3.3, sec3.7, sec3.10
Model-dependent TGX-221 and temozolomide interaction
Cell-based study · MGMT-deficient PI3Kβ-high SF295/U87MG and PI3Kβ-low LN229/A172 glioblastoma cells; human astrocytes. [2]
TGX-221 combined with temozolomide reduced MTS viability more strongly in the PI3Kβ-high glioblastoma models. Single-agent and combination controls were tested together. Low-PI3Kβ cells lacked appreciable positive Bliss interaction, and the tested combination did not significantly alter astrocyte viability.
Literature-Reported Treatment Conditions
- Treatment concentration
- 20 μM TGX-221; 50 μM temozolomide
- Exposure duration
- 4 days
- Method
- MTS viability normalized to vehicle control; Bliss independence interaction analysis.
- Vehicle
- DMSO
- Co-treatment
- TGX-221 plus temozolomide; each agent alone and DMSO controls.
- Reported limits
- Final DMSO percentage not stated in the scoped treatment description.
Methods and source locations
Compound reported in the paper: TGX-221
Evidence for the named research compound in the reported model; no claim of Adooq lot equivalence or clinical efficacy.
Figure 5B image/caption; Results sec2.5; STAR Methods sec5.1, sec5.4.5, sec5.4.6, sec5.5
Combination interaction in SF295 cells: Figure 5B printed EOB annotation and Results sec2.5; EOB is a derived drug-interaction score, not percent tumor inhibition. — Figure 5B image/caption; Results sec2.5; STAR Methods sec5.1, sec5.4.5, sec5.4.6, sec5.5
Combination interaction in U87MG cells: Figure 5B printed EOB annotation and Results sec2.5. — Figure 5B image/caption; Results sec2.5; STAR Methods sec5.1, sec5.4.5, sec5.4.6, sec5.5
Combination interaction in LN229/A172; astrocyte viability: Figure 5B viewed; means and SD from three to four independent replicates per caption. Exact low-model EOB values not adopted because text and figure differ. — Figure 5B image/caption; Results sec2.5; STAR Methods sec5.1, sec5.4.5, sec5.4.6, sec5.5
