AUTHOR=Luo Jinque , Wang Ling , Zhang Li , Tang Wenyu , Deng Meijing , Shi Yuxin , Xu Kun , Wu Qingjun , Zhao Jieyang , Zhang Jinghuan , Li Xin 

TITLE=Targeting adipocyte differentiation with CRT0066101: activation of AMPK signaling in 3T3-L1 cells

JOURNAL=Frontiers in Pharmacology

VOLUME=Volume 16 - 2025

YEAR=2025

URL=https://www.frontiersin.org/journals/pharmacology/articles/10.3389/fphar.2025.1645587

DOI=10.3389/fphar.2025.1645587

ISSN=1663-9812

ABSTRACT=IntroductionObesity is characterized by excessive fat accumulation resulting from adipocyte hypertrophy and hyperplasia, with adipocyte differentiation being a central process driving these changes.MethodsThe anti-adipogenic effects of CRT0066101 (CRT), a pan-protein kinase D (PKD) inhibitor, were evaluated in 3T3-L1 adipocytes. Potential drug targets of CRT were predicted using network pharmacology analysis. The expression of adipocyte-specific genes and proteins was assessed by western blotting and qRT-PCR. To examine the involvement of the AMPK pathway, cells were co-treated with CRT and the AMPK inhibitor Compound C.ResultsCRT significantly inhibited early-stage adipocyte differentiation, reduced lipid accumulation, and downregulated key adipogenic transcription factors, including PPARγ and C/EBPα. Mechanistically, CRT activated the AMPK pathway, a known negative regulator of adipocyte differentiation. Network pharmacology analysis further supported the involvement of AMPK in CRT’s anti-adipogenic action.DiscussionThese findings identify CRT as a novel modulator of adipocyte differentiation through AMPK activation and highlight its potential as a therapeutic candidate for obesity and metabolic syndrome.