Free perindopril erbumine comparator in an ACE substrate-turnover assay
Biochemical assay · Cell-free ACE/hippuryl-histidyl-leucine assay; ACE supplied by Sigma; enzyme species and preparation not specified in the assay paragraph. [1]
The free-erbumine comparator showed high reported ACE inhibition at all three exposure times. The matched nanocomposites were less inhibitory at these times, and the authors reported no inhibition by unloaded Zn/Al layered double hydroxide. Only the free-drug result is adopted. The assay does not establish the molecular species responsible for inhibition.
| Endpoint | Reported result | Reference |
|---|---|---|
| ACE inhibition at 30, 60 and 90 minutes, respectively | ACE inhibition (%) 90.8; 93.6; 97.3 | [1] |
Literature-Reported Treatment Conditions
- Treatment concentration
- 0.5 μg/mL (reported assay concentration)
- Exposure duration
- 20-minute preincubation with ACE; subsequent 30-, 60- or 90-minute incubation at 37°C
- Method
- ACE substrate-turnover assay in pH 8.3 buffer; 3.5 mM substrate solution added after preincubation.
- Co-treatment
- None for the adopted free-PE comparator; PZAE/PZAC formulation groups are not adopted.
- Reported limits
- ACE replicate count, error-bar definition, detailed baseline normalization and drug solvent are not specified in the acquired ACE assay paragraph. No numerical uncertainty is assigned. The triplicate statement occurs in the MTT paragraph and is not transferred to ACE. No assay IC50 or perindoprilat analysis is reported in the selected assay.
Methods and source locations
Compound reported in the paper: Perindopril erbumine; Compound label in the paper: PE (free drug, not PZAE or PZAC)
Applies to the reported free-erbumine assay only. No salt conversion, metabolite potency transfer, nanocomposite equivalence or Adooq lot validation is claimed.
Materials; MTT cytotoxicity and in vitro antihypertensive assay (ACE paragraph); Results: MTT cytotoxicity and in vitro antihypertensive activity (ACE paragraph); Table 5 PE column; Figure 10 and caption
ACE inhibition at 30, 60 and 90 minutes, respectively: Hippuryl-histidyl-leucine conversion to hippuric acid, measured at 228 nm; Table 5 provides exact values; Figure 10 identifies the plotted endpoint as inhibition. — Table 5, PE column, 30/60/90-minute rows; Figure 10 axis and caption; ACE assay methods paragraph