Identification of TG100-115 as a new and potent TRPM7 kinase inhibitor, which suppresses breast cancer cell migration and invasion.
Transient receptor potential melastatin 7 (TRPM7) regulates breast cancer cell proliferation, migration, invasion and metastasis in its ion channel- and kinase domain-dependent manner. The pharmacological effects of TRPM7 ion channel inhibitors on breast cancer cells have been studied, but little is known about the effects of TRPM7 kinase domain inhibitors due to lack of potent TRPM7 kinase inhibitors.



Screening was performed by using TRPM7 kinase assay. Effects of TG100-115 on breast cancer cell proliferation, migration, invasion, myosin IIA phosphorylation, and TRPM7 ion channel activity were assessed by using MTT, wound healing, transwell assay, Western blotting, and patch clamping, respectively.



We found that CREB peptide is a potent substrate for the TR-FRET based TRPM7 kinase assay. Using this method, we discovered a new and potent TRPM7 kinase inhibitor, TG100-115. TG100-115 inhibited TRPM7 kinase activity in an ATP competitive fashion with over 70-fold stronger activity than that of rottlerin, known as a TRPM7 kinase inhibitor. TG100-115 has little effect on proliferation of MDA-MB-231 cells, but significantly decreases cell migration and invasion. Moreover, TG100-115 inhibits TRPM7 kinase regulated phosphorylation of the myosin IIA heavy chain and phosphorylation of focal adhesion kinase. TG100-115 also suppressed TRPM7 ion channel activity.



TG100-115 can be used as a potent TRPM7 kinase inhibitor and a potent inhibitor of breast cancer cell migration.



TG100-115 could be a useful tool for studying the pharmacological effects of TRPM7 kinase activity aimed at providing insight into new therapeutic approaches to the treatment of breast cancer.
Chemical Kinomics Research Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea.
Chiman
C
Song
School of Biosystem and Biomedical Science, College of Health Science, Korea University, Seoul, 02841, Republic of Korea.
Yeonju
Y
Bae
Chemical Kinomics Research Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea.
JinJoo
J
Jun
Chemical Kinomics Research Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; Department of Biological Chemistry, Korea University of Science and Technology, Daejeon 34113, Republic of Korea.
Hyomin
H
Lee
New Drug Development Center, Daegu-Gyeongbuk Medical Innovation Foundation, Daegu 41061, Republic of Korea.
Nam Doo
ND
Kim
Division of Life Science, Korea Basic Science Institute, Daejeon 34133, Republic of Korea.
Kyung-Bok
KB
Lee
Chemical Kinomics Research Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; Department of Biological Chemistry, Korea University of Science and Technology, Daejeon 34113, Republic of Korea.
Wooyoung
W
Hur
School of Biosystem and Biomedical Science, College of Health Science, Korea University, Seoul, 02841, Republic of Korea.
Jae-Yong
JY
Park
Chemical Kinomics Research Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; KU-KIST Graduate School of Converging Science and Technology, Korea University, Seoul 02841, Republic of Korea. Electronic address: tbsim@kist.re.kr.
Taebo
T
Sim
D000255:Adenosine Triphosphate / Q000378:metabolism; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D005260:Female; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism; D006801:Humans; D009361:Neoplasm Invasiveness / Q000473:pathology*; D024507:Nonmuscle Myosin Type IIA / Q000378:metabolism; D010636:Phenols / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors*; D011621:Pteridines / Q000494:pharmacology*; D050053:TRPM Cation Channels / Q000037:antagonists & inhibitors*
C539252:3-(2,4-diamino-6-(3-hydroxyphenyl)pteridin-7-yl)phenol; D017362:Cyclic AMP Response Element-Binding Protein; D010636:Phenols; D047428:Protein Kinase Inhibitors; D011621:Pteridines; D050053:TRPM Cation Channels; D000255:Adenosine Triphosphate; D051416:Focal Adhesion Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; C486583:TRPM7 protein, human; D024507:Nonmuscle Myosin Type IIA
Breast cancer; Cell invasion; Cell migration; MDA-MB-231; TG100-115; TRPM7 kinase inhibitor