In primary mouse cortical neurons exposed to 1.5 h of oxygen-glucose deprivation, P7C3-A20 added at injury onset improved viability at 30 μM. This result is specific to the brief OGD model. [3]
Responses varied with neuronal preparation and exposure conditions. In a separate study, P7C3-A20 partially rescued doxorubicin-depleted NAD in primary cortical neurons but not superior cervical ganglion explants. The same study observed concentration-dependent spontaneous neurite degeneration during 24 h exposure, beginning at 500 nM in the tested preparations. These findings limit generalization of neuronal protection or tolerability across models. [4]
Reported stimulation in a coupled NAMPT assay
Biochemical assay · Purified recombinant human NAMPT coupled to NMNAT and alcohol dehydrogenase [1]
P7C3-A20 increased the coupled reaction rate with increasing concentration. The main text reports no enhancement when NAMPT was omitted. This is an assay-specific observation, not an established binding site or an EC50.
Literature-Reported Treatment Conditions
- Treatment concentration
- 0.3, 1 and 3 μM
- Exposure duration
- Kinetic traces over approximately 0-4 min (Figure 7A)
- Method
- Spectrophotometric NADH appearance at OD340 nm
- Vehicle
- Compounds in DMSO further diluted in ethanol; control reaction measured before compound addition
- Reported limits
- No fitted activation potency adopted. The specificity-control figure S6 was not retrieved; its result is reported in the acquired main Results text.
Methods and source locations
Compound reported in the paper: P7C3-A20; Compound label in the paper: P7C3-A20
Applies to the reported P7C3-A20 material and assay; no salt conversion, enantiomer-specific transfer, or Adooq lot validation.
Figure 7A and caption; Results: Active variants enhance purified NAMPT; Experimental procedures: In vitro NAMPT assay
Coupled NAMPT reaction rate: NAMPT-dependent NMN formation coupled through NMNAT and alcohol dehydrogenase to NADH detection — Figure 7A and caption; Results paragraph immediately preceding Figure 7
No detectable displacement in a NAMPT binding assay
Biochemical assay · NAMPT fluorescence-polarization probe-displacement assay using ZN-2-102 [2]
P7C3-A20 showed no detectable NAMPT binding activity in this assay. FK866 and NP-A1R displaced the probe. Failure to displace this probe does not exclude every possible mode of interaction with NAMPT.
Literature-Reported Treatment Conditions
- Method
- Fluorescence polarization / probe displacement
- Reported limits
- Detailed supplemental assay conditions, compound sourcing and replicate information were not retrieved. No numerical affinity or universal absence of binding is asserted.
Methods and source locations
Compound reported in the paper: P7C3-A20; Compound label in the paper: P7C3-A20
Applies to the reported P7C3-A20 material and assay; no salt conversion, enantiomer-specific transfer, or Adooq lot validation.
Figure 3A-C and caption; Results & Discussion: N-PAM binding to the NAMPT rear channel
Displacement of NAMPT-bound FP probe: ZN-2-102 probe displacement; positive displacement controls in Figure 3A — Figure 3B-C; Results paragraph following Figure 3
Protection during brief oxygen-glucose deprivation
Cell-based study · Primary cortical neurons from embryonic day 18 mouse cortex, cultured for 7 days [3]
Treatment at OGD onset increased viability at 30 μM compared with OGD vehicle. Normal and OGD vehicle controls were included; Tuj-1/DAPI staining provided supporting evidence. This brief-injury result does not establish general neuronal tolerability.
Literature-Reported Treatment Conditions
- Treatment concentration
- 3, 10 and 30 μM
- Exposure duration
- 1.5 h OGD; viability measured immediately afterward
- Method
- Addition at the beginning of OGD
- Vehicle
- DMSO; final DMSO ≤1‰
- Reported limits
- n=4 for viability; the caption does not define whether these are independent cultures. Longer OGD and reperfusion experiments are read but not separate adopted studies.
Methods and source locations
Compound reported in the paper: P7C3-A20; Compound label in the paper: P7C3-A20
Applies to the reported P7C3-A20 material and assay; no salt conversion, enantiomer-specific transfer, or Adooq lot validation.
Figure 2A-C; Results: P7C3-A20 Protects Primary Cortical Neurons against OGD-Induced Neuronal Injury; corresponding Methods
Cell viability at 30 μM relative to normal conditions: CCK-8 assay; figure expresses viability as fold of normal — Figure 2A; Results paragraph describing the 1.5-h OGD dose-response experiment; Statistical Analysis
Concentration-dependent neurite degeneration
Cell-based study · Murine dissociated superior cervical ganglion neurons, superior cervical ganglion explants and primary cortical neuronal cultures [4]
P7C3-A20 caused concentration-dependent spontaneous neurite degeneration. Concentrations of 100 nM or below did not cause toxicity at 24 h in these conditions; degeneration occurred at 500 nM or greater. This concentration window is specific to these preparations.
Literature-Reported Treatment Conditions
- Treatment concentration
- 50 nM, 100 nM, 500 nM, 5 μM and 10 μM
- Exposure duration
- 24 h
- Method
- Exposure followed by neurite degeneration-index imaging
- Vehicle
- Control included; DMSO concentration not specified in the acquired Figure 2 description
- Reported limits
- No precise degeneration-index values estimated from graph. The repeat designation is preserved as reported.
Methods and source locations
Compound reported in the paper: P7C3-A20; Compound label in the paper: P7C3-A20
Applies to the reported P7C3-A20 material and assay; no salt conversion, enantiomer-specific transfer, or Adooq lot validation.
Figure 2A-C and caption; first Results paragraph; P7C3-A20, Neurite degeneration index and Statistics Methods
Spontaneous neurite degeneration: Operator-independent ImageJ-based fragmentation index comparing baseline and 24 h — Figure 2A-C and caption; Results first paragraph; Conclusion
NAD rescue depends on neuronal preparation
Cell-based study · Murine primary cortical neurons (PCN) and superior cervical ganglion explants (SCGe) [4]
P7C3-A20 partially rescued doxorubicin-depleted NAD in PCN but did not significantly rescue NAD in SCGe. This assay measured the NAD/NADH pool by luminescence and did not directly assay NAMPT catalysis or neuronal survival.
Literature-Reported Treatment Conditions
- Treatment concentration
- 100 nM P7C3-A20
- Exposure duration
- 48 h
- Method
- Co-treatment with doxorubicin; control, P7C3-A20 alone, doxorubicin alone and combination groups
- Vehicle
- DMSO control; final vehicle concentration not specified in the acquired main text
- Co-treatment
- 500 nM doxorubicin
- Reported limits
- No absolute NAD concentrations inferred from luminescence. No numerical between-study potency comparison is justified.
Methods and source locations
Compound reported in the paper: P7C3-A20; Compound label in the paper: P7C3-A20
Applies to the reported P7C3-A20 material and assay; no salt conversion, enantiomer-specific transfer, or Adooq lot validation.
Figure 4A-B and caption; NAD-rescue Results paragraph; NAD/NADH-Glo and Statistics Methods
NAD/NADH-pool rescue during doxorubicin exposure: NAD/NADH-Glo assay, PHERAstar luminescence reading — Figure 4A-B and caption; Results paragraph on doxorubicin-induced depletion
