In primary human hepatocytes, PSI-6130 exposure generated triphosphate metabolites with different persistence after washout. Reported intracellular half-lives were 4.7 ± 0.6 h for PSI-6130-TP and 38 ± 16 h for RO2433-TP (mean ± SD; four donors). These are cellular metabolite half-lives. [2]
Host-kinase activation of PSI-6130
Biochemical assay · Purified recombinant human dCK, UCK-1, UMP-CMP kinase (YMPK), and NDPK [1]
Comparative substrate assays support sequential phosphorylation through dCK, YMPK and NDPK. UCK-1 did not phosphorylate PSI-6130 under the tested conditions.
Literature-Reported Treatment Conditions
- Method
- Cell-free coupled spectrophotometric substrate comparison
- Reported limits
- No treatment-dose interpretation. Replicate count not specified in the retrieved table footnotes; no numeric dCK turnover or catalytic-efficiency claim adopted.
Methods and source locations
Compound reported in the paper: PSI-6130 and its phosphorylated derivatives
Research findings for the stated material and model; no Adooq lot validation.
Other materials; dCK/UCK-1/YMPK/NDPK assays; Results, Tables 1–4; Figure 1
Parent nucleoside phosphorylation: Coupled spectrophotometric assay; cytidine and uridine were active UCK-1 comparators. — Results: Phosphorylation by dCK and UCK-1; Tables 1–2
Sequential MP-to-DP and DP-to-TP phosphorylation: Natural nucleotide comparators; NDPK-free coupled-assay background control. — Results: Phosphorylation by YMPK and NDPK; Tables 3–4
Chain termination by PSI-6130 triphosphate
Biochemical assay · Purified HCV NS5B polymerase; cell-free RNA extension assay [1]
PSI-6130-TP produced an RNA six-mer termination pattern comparable to 3′-dCTP, whereas the natural-nucleotide control yielded elongated products.
Literature-Reported Treatment Conditions
- Treatment concentration
- 100 μM PSI-6130-TP
- Exposure duration
- 1 h
- Method
- Cell-free polymerase reaction at 27 °C; sequencing-gel endpoint
- Reported limits
- No parent-nucleoside dose or cellular potency inferred; replicate count not specified in the retrieved Figure 2 caption.
Methods and source locations
Compound reported in the paper: PSI-6130-TP
Research findings for the stated material and model; no Adooq lot validation.
Chain-termination study; Results: Chain termination; Figure 2 and caption
RNA extension after analog incorporation: Phosphorimaging; natural CTP control and known chain terminator 3′-dCTP comparator. — Figure 2, lanes 1, 2 and 4; Results: Chain termination
Persistence of intracellular triphosphate metabolites
Cell-based study · Primary human hepatocytes from four donors; noninfected metabolism model [2]
After PSI-6130 exposure and extracellular washout, the uridine triphosphate metabolite RO2433-TP persisted longer than PSI-6130-TP.
Literature-Reported Treatment Conditions
- Treatment concentration
- 2 μM PSI-6130
- Exposure duration
- 24 h exposure; follow-up to 72 h after extracellular removal
- Method
- Radiotracer HPLC with reference standards; duplicate cell samples per time point
- Reported limits
- Final exposure vehicle composition not fully specified; PBS parent stock and ethanol tracer stock are described, without a final vehicle percentage. Untreated controls and viable-cell counts described.
Methods and source locations
Compound reported in the paper: PSI-6130
Research findings for the stated material and model; no Adooq lot validation.
Experimental Procedures pp.29813–14; Intracellular Stability Results p.29817; Figure 6 and Table 4 p.29818
Intracellular PSI-6130-TP decay after parent washout: Nonlinear single-phase exponential fit; half-life relative to highest post-removal triphosphate level. — Table 4 and footnote, p.29818; Intracellular Stability Results, p.29817
Intracellular RO2433-TP decay after parent washout: Same washout experiment; Table 4 rounding preserved. — Table 4 and footnote, p.29818
