Cimicifuga heracleifolia is a perennial herb that belongs to the Ranunculaceae family. Its dried rhizomes are a source of Cimicifugae Rhizoma, a traditional Chinese medicine used for detoxification, the treatment of febrile diseases, and the prevention of pathogenic invasion. In the present study,
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Cimicifuga heracleifolia is a perennial herb that belongs to the Ranunculaceae family. Its dried rhizomes are a source of Cimicifugae Rhizoma, a traditional Chinese medicine used for detoxification, the treatment of febrile diseases, and the prevention of pathogenic invasion. In the present study, a phytochemical investigation of the rhizomes of
C. heracleifolia resulted in the isolation of three indole alkaloids (
1–
3) and seven phenolic amides (
4–
10), including three new compounds, 6-methoxy-3-(3-methyl-1-oxo-2-butenyl) 1
H indole (
1), (3
R)-1-(
β-
d-glucopyranosyl)-3-hydroxy-3-(3-methylbutyl)-2-oxindole (
3), and
N-acetyldopamine-3-
O-
β-
d-allopyranoside (
4). Their structures were elucidated using extensive physicochemical and spectroscopic analyses. All the isolated compounds were evaluated for their inhibitory activity against soluble epoxide hydrolase (sEH). The results showed that cimicifugamide A (
6) exhibited the most potent inhibitory activity, with an IC₅₀ value of 8.74 μM, followed by cimicifugamide (
7), demethoxycimicifugamide (
8), and
N-
trans-feruloyl tyramine (
10), with IC
50 values ranging from 15.63 to 20.58 μM. Kinetic analysis revealed that compound
6 inhibited sEH through a non-competitive mechanism.
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