EVP4593 (300 nM) suppressed store-operated calcium-associated influx in YAC128 mouse striatal neurons, with no significant suppression in wild-type neurons in that assay. In corticostriatal cultures, 30 nM treatment for 16 h restored YAC128 spine density toward wild-type control levels but reduced spine density in wild-type cultures. [1,3]
Partial inhibition of stimulated NF-κB reporter activity
Cell-based study · Human Jurkat E6-1 cells carrying an NF-κB luciferase reporter [1]
EVP4593 partially suppressed PMA/PHA-induced reporter activity. The structurally unrelated IKK inhibitor BMS-345541 showed fuller inhibition. This is pathway-level activity, not evidence of direct binding to NF-κB.
Literature-Reported Treatment Conditions
- Treatment concentration
- 1 µM
- Exposure duration
- 30 min pretreatment; 6 h after stimulation
- Method
- Luciferase reporter after PMA 50 ng/ml plus PHA 100 µg/ml stimulation
- Vehicle
- DMSO stocks; final vehicle fraction for this reporter assay not specified in the read methods
- Co-treatment
- PMA/PHA are the activating stimuli; BMS-345541 and analogues are separate comparator conditions.
- Reported limits
- No compound-specific numerical MTS viability result is provided in the selected main text; MTS viability testing is described. No direct target-binding measurement.
Methods and source locations
Compound reported in the paper: QNZ (EVP4593); Compound label in the paper: EVP4593
Evidence concerns the named/depicted compound in the reported preparation; salt, purity and lot equivalence to Adooq A12748 are not established.
Figure 1C–D and associated Results; Experimental Procedures: Compounds and NF-κB assays
PMA/PHA-induced NF-κB reporter activity: Bright-Glo luciferase assay; magnitude is the authors’ reported range, not a fitted IC50 or an exact value digitized from the bar — Figure 1C–D and associated Results; Experimental Procedures: Compounds and NF-κB assays
SOC-associated calcium influx in primary striatal neurons
Cell-based study · Wild-type and YAC128 mouse primary medium spiny neurons, DIV10–14 [1]
At 300 nM, EVP4593 reduced the calcium add-back response in YAC128 neurons. The same concentration had no significant effect in wild-type neurons; inactive analogue EVP14808 did not suppress the YAC128 response.
Literature-Reported Treatment Conditions
- Treatment concentration
- 300 nM
- Exposure duration
- Added 10 min before calcium readdition
- Method
- Fura-2 340/380 imaging after store depletion with thapsigargin (1 µM, 5 min), then calcium readdition (2 mM)
- Vehicle
- DMSO control; assay-specific final fraction not specified in the read calcium-imaging methods
- Co-treatment
- Thapsigargin is used for store depletion; active analogue EVP14756 and inactive EVP14808 are separate comparator conditions.
- Reported limits
- n counts cells; distinct donor/culture replication for each Figure 2 group is not tabulated. General methods state experiments repeated at least three times.
Methods and source locations
Compound reported in the paper: QNZ (EVP4593); Compound label in the paper: EVP4593
Evidence concerns the named/depicted compound in the reported preparation; salt, purity and lot equivalence to Adooq A12748 are not established.
Figure 2A–E and associated Results; Experimental Procedures: Compounds, SOC measurements and Statistical analysis
Calcium add-back response in YAC128 MSNs: Average increase in Fura-2 ratio; exact values from Results, corroborated by Figure 2E — Figure 2A–E and associated Results; Experimental Procedures: Compounds, SOC measurements and Statistical analysis
Wild-type response and inactive-analogue control: Figure 2E and Results; no exact effect size inferred from bars — Figure 2A–E and associated Results; Experimental Procedures: Compounds, SOC measurements and Statistical analysis
Complex I enzymatic inhibition in mitochondrial membranes
Biochemical assay · Mitochondrial membrane preparations from Yarrowia lipolytica and bovine heart [2]
EVP4593 inhibited complex I in both preparations with nanomolar half-maximal inhibitory concentrations. These biochemical assay potencies are distinct from cellular reporter potencies.
Literature-Reported Treatment Conditions
- Treatment concentration
- Concentration-response titration; no exact series transcribed from the plotted points
- Exposure duration
- Preincubation described; duration not available in retrieved main text
- Method
- dNADH:quinone catalytic activity, absorbance monitoring of dNADH oxidation after decylubiquinone addition
- Vehicle
- Not specified for the selected enzyme assay in retrieved main text
- Co-treatment
- DQA is the positive-control inhibitor in the functional screening assay, not a combined EVP4593 treatment.
- Reported limits
- SI not acquired; detailed enzyme-assay recipe, preincubation interval, replicate number and definition of ± for Figure 5 remain unavailable in the read main text.
Methods and source locations
Compound reported in the paper: QNZ (EVP4593); Compound label in the paper: EVP4593
Evidence concerns the named/depicted compound in the reported preparation; salt, purity and lot equivalence to Adooq A12748 are not established.
Results and discussion, Figure 2 compound structure and Figure 5 with caption; main-text enzymatic-assay description
Complex I activity in Y. lipolytica membranes: Concentration-response inhibition; Figure 5 red circles and Results — Results and discussion, Figure 2 compound structure and Figure 5 with caption; main-text enzymatic-assay description
Complex I activity in bovine heart mitochondrial membranes: Concentration-response inhibition; Figure 5 black circles and Results — Results and discussion, Figure 2 compound structure and Figure 5 with caption; main-text enzymatic-assay description
Complex I-dependent respiration and succinate rescue
Biochemical assay · Intact isolated rat heart mitochondria [2]
EVP4593 inhibited malate/glutamate-supported ADP-stimulated respiration. Subsequent succinate addition restored respiration, supporting a complex I-associated effect in this preparation. This bypass control does not constitute an exhaustive selectivity panel.
Literature-Reported Treatment Conditions
- Treatment concentration
- Increasing concentrations shown in Figure 6A: 5, 10, 20, 30, 40 and 140 nM
- Exposure duration
- Stepwise titration after stable state 3 oxygen consumption; per-step times not specified in retrieved main text
- Method
- High-resolution respirometry at 37 °C (Oxygraph-2k)
- Vehicle
- Not specified for Figure 6 in retrieved main text
- Co-treatment
- 5 mM malate plus 5 mM glutamate and 2 mM ADP; subsequent 5 mM succinate bypass control; final 2 mM KCN respiratory inhibition control
- Reported limits
- Replicate number, uncertainty semantics and detailed fitting model not specified in retrieved main text; SI not acquired.
Methods and source locations
Compound reported in the paper: QNZ (EVP4593); Compound label in the paper: EVP4593
Evidence concerns the named/depicted compound in the reported preparation; salt, purity and lot equivalence to Adooq A12748 are not established.
Results and discussion, Figure 6A–B and caption
Complex I-dependent state 3 oxygen consumption: Malate/glutamate-supported oxygen flux; Figure 6B and associated Results — Results and discussion, Figure 6A–B and caption
Respiratory bypass after EVP4593 inhibition: Figure 6A; qualitative direction visually checked, no exact oxygen-flux values digitized — Results and discussion, Figure 6A–B and caption
Genotype-dependent spine responses in corticostriatal cultures
Cell-based study · Wild-type and YAC128 mouse corticostriatal cocultures; medium spiny neurons [3]
At 30 nM, EVP4593 restored spine density in YAC128 cultures toward the wild-type control level but reduced density in wild-type cultures. These opposite responses limit broad neuroprotection claims.
Literature-Reported Treatment Conditions
- Treatment concentration
- 30 nM
- Exposure duration
- 16 h starting at DIV20; fixed at DIV21
- Method
- DARPP32 staining, confocal imaging and NeuronStudio spine quantification
- Vehicle
- Vehicle-only controls; culture vehicle composition not specified for Figure 11A–B in the read text
- Co-treatment
- Single-compound treatment; genotype-matched vehicle controls
- Reported limits
- Vehicle composition for culture treatment not specified; Figure 11 caption reports n = 15 per condition without explicitly defining the sampling unit. General methods describe at least three culture batches.
Methods and source locations
Compound reported in the paper: QNZ (EVP4593); Compound label in the paper: EVP4593
Evidence concerns the named/depicted compound in the reported preparation; salt, purity and lot equivalence to Adooq A12748 are not established.
Figure 11A–B, associated Results; Materials and Methods: Dendritic spine analysis of MSNs in primary corticostriatal cocultures; Figure 10A identity
YAC128 MSN spine density: DARPP32-positive MSN spines; Figure 11A–B and Results — Figure 11A–B, associated Results; Materials and Methods: Dendritic spine analysis of MSNs in primary corticostriatal cocultures; Figure 10A identity
Wild-type MSN spine density: Figure 11A–B and Results; unfavorable effect retained — Figure 11A–B, associated Results; Materials and Methods: Dendritic spine analysis of MSNs in primary corticostriatal cocultures; Figure 10A identity
