Biological evaluation of tanshindiols as EZH2 histone methyltransferase inhibitors.
EZH2 is the core subunit of Polycomb repressive complex 2 catalyzing the methylation of histone H3 lysine-27 and closely involved in tumorigenesis. To discover small molecule inhibitors for EZH2 methyltransferase activity, we performed an inhibitor screen with catalytically active EZH2 protein complex and identified tanshindiols as EZH2 inhibitors. Tanshindiol B and C potently inhibited the methyltransferase activity in in vitro enzymatic assay with IC50 values of 0.52μM and 0.55μM, respectively. Tanshindiol C exhibited growth inhibition of several cancer cells including Pfeiffer cell line, a diffuse large B cell lymphoma harboring EZH2 A677G activating mutation. Tanshindiol treatment in Pfeiffer cells significantly decreased the tri-methylated form of histone H3 lysine-27, a substrate of EZH2, as revealed by Western blot analysis and histone methylation ELISA. Based on enzyme kinetics and docking studies, we propose that tanshindiol-mediated inhibition of EZH2 activity is competitive for the substrate S-adenosylmethionine. Taken together, our findings strongly suggest that tanshindiols possess a unique anti-cancer activity whose mechanism involves the inhibition of EZH2 activity and would provide chemically valuable information for designing a new class of potent EZH2 inhibitors.
Pharmacology Research Group, Drug Discovery Division, Korea Research Institute of Chemical Technology, Gajeong-Ro 141, Yuseong-gu, Daejeon 305-600, Republic of Korea; Medicinal and Pharmaceutical Chemistry, University of Science and Technology, Gajeong-Ro 217, Yuseong-gu, Daejeon 305-350, Republic of Korea.
Jimin
J
Woo
Pharmacology Research Group, Drug Discovery Division, Korea Research Institute of Chemical Technology, Gajeong-Ro 141, Yuseong-gu, Daejeon 305-600, Republic of Korea.
Hyun-Young
HY
Kim
Pharmacology Research Group, Drug Discovery Division, Korea Research Institute of Chemical Technology, Gajeong-Ro 141, Yuseong-gu, Daejeon 305-600, Republic of Korea.
Byung Jin
BJ
Byun
Pharmacology Research Group, Drug Discovery Division, Korea Research Institute of Chemical Technology, Gajeong-Ro 141, Yuseong-gu, Daejeon 305-600, Republic of Korea.
Chong-Hak
CH
Chae
Pharmacology Research Group, Drug Discovery Division, Korea Research Institute of Chemical Technology, Gajeong-Ro 141, Yuseong-gu, Daejeon 305-600, Republic of Korea; Graduate School of New Drug Discovery and Development, Chungnam National University, Daehak-ro 79, Yuseong-gu, Daejeon 305-764, Republic of Korea.
Ji Young
JY
Lee
Pharmacology Research Group, Drug Discovery Division, Korea Research Institute of Chemical Technology, Gajeong-Ro 141, Yuseong-gu, Daejeon 305-600, Republic of Korea; Graduate School of New Drug Discovery and Development, Chungnam National University, Daehak-ro 79, Yuseong-gu, Daejeon 305-764, Republic of Korea.
Shi Yong
SY
Ryu
Pharmacology Research Group, Drug Discovery Division, Korea Research Institute of Chemical Technology, Gajeong-Ro 141, Yuseong-gu, Daejeon 305-600, Republic of Korea.
Woo-Kyu
WK
Park
Pharmacology Research Group, Drug Discovery Division, Korea Research Institute of Chemical Technology, Gajeong-Ro 141, Yuseong-gu, Daejeon 305-600, Republic of Korea; Medicinal and Pharmaceutical Chemistry, University of Science and Technology, Gajeong-Ro 217, Yuseong-gu, Daejeon 305-350, Republic of Korea. Electronic address: hycho@krict.re.kr.
Heeyeong
H
Cho
Pharmacology Research Group, Drug Discovery Division, Korea Research Institute of Chemical Technology, Gajeong-Ro 141, Yuseong-gu, Daejeon 305-600, Republic of Korea; Medicinal and Pharmaceutical Chemistry, University of Science and Technology, Gajeong-Ro 217, Yuseong-gu, Daejeon 305-350, Republic of Korea. Electronic address: gchoi@krict.re.kr.
Gildon
G
Choi
D045784:Abietanes / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor; D000071221:Enhancer of Zeste Homolog 2 Protein; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006801:Humans; D008968:Molecular Conformation; D063151:Polycomb Repressive Complex 2 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013329:Structure-Activity Relationship
D045784:Abietanes; D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D000076983:Histone Methyltransferases; C498409:EZH2 protein, human; D000071221:Enhancer of Zeste Homolog 2 Protein; D011495:Histone-Lysine N-Methyltransferase; D063151:Polycomb Repressive Complex 2
EZH2; Histone methyltransferase; Tanshindiol; Tanshinone