Identification of cepharanthine as a novel senolytic that selectively eliminates senescent cells by regulating cell survival signaling pathways

Senolytics are small-molecule drugs that selectively induce apoptosis in senescent cells, thereby reducing the chronic inflammation and tissue damage caused by the senescence-associated secretory phenotype (SASP) and delaying aging, improving organ function and extending healthspan. Existing senolytics, however, often suffer from low bioavailability and off-target toxicity, so potent and safer senolytics are urgently needed.

To screen for novel senolytic compounds and validate the ability of cepharanthine (CEP) to eliminate senescent cells, improve organ function and extend lifespan.

Preclinical study combining in vitro phenotypic screening and mechanistic assays with in vivo validation in zebrafish, C. elegans, naturally aged mice and a Hutchinson-Gilford progeria syndrome (HGPS) mouse model.

Senolytic efficacy was assessed by a senescent cell-based phenotypic screen, IC₅₀ determination, inhibitor-based rescue assays and multiple senescence models. RNA-seq and pathway enrichment identified the key PI3K/AKT and PERK-ATF4-CHOP pathways, verified by Western blot, qRT-PCR, immunofluorescence and mitochondrial assays. In vivo efficacy was evaluated by senescence-associated β-galactosidase (SA-β-gal) staining, HE staining, immunofluorescence, ELISA, biochemical assays and survival analysis.

CEP selectively eliminates senescent cells by cooperatively and in parallel regulating the PI3K/AKT and PERK-ATF4-CHOP pathways, differentially inducing PUMA/NOXA and triggering mitochondrial intrinsic apoptosis. In zebrafish, C. elegans, naturally aged mice and HGPS mice, CEP clears senescent cells, reduces SASP levels, improves organ function and prolongs lifespan.

CEP is a novel natural senolytic that induces mitochondrial intrinsic apoptosis via parallel, non-redundant regulation of the PI3K/AKT and PERK-ATF4-CHOP pathways, showing good potential for clinical translation.