The inherent structural instability
of some physalins has hampered
the isolation and identification of these compounds for approximately
50 years, and an effective method to overcome these challenges remains
unavailable. In the present study, the unprecedented tautomerization
mechanism of unstable physalins was elucidated by performing isotopic
labeling experiments and DFT calculations, which led to the successful
separation of tautomers and isolation of highly pure products for
the first time. As a result, 15 new physalins, physaminins A–O
(1–15), as well as 17 known analogues
(16–32), were isolated from the whole
plants of Physalis minima L. The chemical structures
of the new compounds were established by performing a comprehensive
analysis of spectroscopic data, and their absolute configurations
were confirmed by using computational ECD calculations and/or single-crystal
X-ray diffraction analyses. All obtained isolates were evaluated for
their antiproliferative effects against four human cancer cell lines
(A549, HepG2, MCF-7, and SCG-7901) and two noncancerous cell lines
(RAW 264.7 and human normal hepatocytes L02), as well as their anti-inflammatory
activities by measuring their abilities to inhibit NO production in
LPS-stimulated murine RAW 264.7 cells in vitro. Compounds 1–5, 13, 16, 18, 19, 23, and 30 exerted
significant antiproliferative effects on the four human cancer lines,
with IC50 values ranging from 0.2(0) to 24.7(2) μM,
and these compounds were not toxic to the two noncancerous cell lines
at a concentration of 10 μM. Moreover, compounds 7, 10, 11, 12, 14, 17, 22, and 27 significantly
inhibited NO production, with IC50 values ranging from
2.9(1) to 9.5(2) μM.