摘要Src homology 2 domain-containing phosphatase 2(SHP2)is a pivotal regulator linking receptor tyrosine kinase(RTK)signaling.Abnormal SHP2 activity has been associated with tumorigenesis and metastasis.Although some SHP2-targeting modulators have entered clinical trials,U.S.Food and Drug Administraction(FDA)-approved SHP2 targeting drugs are still not available.Herein,we describe cooperative biochemical inhibition experiments that facilitate the identification of both catalytic and allosteric SHP2 inhibitors using an in-house natural product(NP)library.Based on this screening methodology,structurally diverse sets of NPs were characterized,among which dihydrotanshinone I(DHT)potently inhibited the wild-type SHP2 protein tyrosine phosphatase(PTP)domain and gain-of-function SHP2 variants.Trichostatin A(TSA)bound to the"tunnel"binding site,acting as an allosteric inhibitor.This study illustrates an optimized screening methodology and tactics to identify novel SHP2 modulators from NPs and provides a foundation for further NP-based drug development for the treatment of RTK-driven cancer.