Ten newDaphniphyllumalkaloids, calycindaphines A-J (1-10), together with seventeen known alkaloids were isolated from the roots ofDaphniphyllum calycinum. Their structures were established by extensive spectroscopic methods and compared with data from literature. Compound1is a novel alkaloid with a new rearrangement C22skeleton with the 5/8/7/5/5 ring system. Compound2represents the second example of calyciphylline G-type alkaloids. Compound10is the first example of secodaphniphylline-type alkaloid absent of the oxygen-bridge between C-25/C-29. The possible biogenetic pathways of1and2were also proposed. All the isolated compounds were evaluated for their bioactivities in three cell models. Compounds22,23, and26showed significant NF-κB transcriptional inhibitory activity at a concentration of 50 μM. Compounds16and18exhibited significant TGF-β inhibitory activity in HepG2 cells. Compounds24and26induced autophagic puncta and mediated the autophagic marker LC3-II conversion in HEK293 cells. © The Royal Society of Chemistry 2021.