Validation and development of MTH1 inhibitors for treatment of cancer.
Previously, we showed cancer cells rely on the MTH1 protein to prevent incorporation of otherwise deadly oxidised nucleotides into DNA and we developed MTH1 inhibitors which selectively kill cancer cells. Recently, several new and potent inhibitors of MTH1 were demonstrated to be non-toxic to cancer cells, challenging the utility of MTH1 inhibition as a target for cancer treatment.



Human cancer cell lines were exposed in vitro to MTH1 inhibitors or depleted of MTH1 by siRNA or shRNA. 8-oxodG was measured by immunostaining and modified comet assay. Thermal Proteome profiling, proteomics, cellular thermal shift assays, kinase and CEREP panel were used for target engagement, mode of action and selectivity investigations of MTH1 inhibitors. Effect of MTH1 inhibition on tumour growth was explored in BRAF V600E-mutated malignant melanoma patient derived xenograft and human colon cancer SW480 and HCT116 xenograft models.



Here, we demonstrate that recently described MTH1 inhibitors, which fail to kill cancer cells, also fail to introduce the toxic oxidized nucleotides into DNA. We also describe a new MTH1 inhibitor TH1579, (Karonudib), an analogue of TH588, which is a potent, selective MTH1 inhibitor with good oral availability and demonstrates excellent pharmacokinetic and anti-cancer properties in vivo.



We demonstrate that in order to kill cancer cells MTH1 inhibitors must also introduce oxidized nucleotides into DNA. Furthermore, we describe TH1579 as a best-in-class MTH1 inhibitor, which we expect to be useful in order to further validate the MTH1 inhibitor concept.
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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Warpman Berglund
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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K
Sanjiv
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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H
Gad
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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Kalderén
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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Koolmeister
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
T
T
Pham
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
C
C
Gokturk
Clinical Proteomics Mass Spectrometry, Department of Oncology-Pathology.
R
R
Jafari
Clinical Proteomics Mass Spectrometry, Department of Oncology-Pathology.
G
G
Maddalo
Chemical Biology Consortium Sweden, Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
B
B
Seashore-Ludlow
Division of Physiological Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
A
A
Chernobrovkin
Division of Physiological Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
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A
Manoilov
Molecular Carcinogenesis Group, Department of Histology and Embryology, School of Medicine, National and Kapodistrian University of Athens, Athens, Greece.
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IS
Pateras
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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Rasti
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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AS
Jemth
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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Almlöf
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
O
O
Loseva
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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Visnes
Sahlgrenska Translational Melanoma Group (SATMEG), Sahlgrenska Cancer Center, Department of Surgery, Institute of Clinical Sciences, University of Gothenburg and Sahlgrenska University Hospital, Gothenburg.
B O
BO
Einarsdottir
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.|Department of Pharmacy and.
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FZ
Gaugaz
Science for Life Laboratory Drug Discovery and Development Platform, ADME of Therapeutics facility, Department of Phamracy, Uppsala University, Uppsala, Sweden.
A
A
Saleh
Swedish Toxicology Sciences Research Center, Södertälje, Sweden.
B
B
Platzack
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
O A
OA
Wallner
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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Vallin
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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M
Henriksson
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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P
Wakchaure
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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S
Borhade
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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Herr
National Bioinformatics Infrastructure Sweden (NBIS), Science for Life Laboratory, Department of Medicine Solna, Karolinska Institutet, Stockholm.
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Y
Kallberg
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.|Science for Life Laboratory Drug Discovery and Development Platform, ADME of Therapeutics facility, Department of Phamracy, Uppsala University, Uppsala, Sweden.
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P
Baranczewski
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
E J
EJ
Homan
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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E
Wiita
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.|SP Process Development, Södertälje, Sweden.
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Nagpal
SP Process Development, Södertälje, Sweden.
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Meijer
SP Process Development, Södertälje, Sweden.
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Schipper
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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Rudd
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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Bräutigam
Science for Life Laboratory Drug Discovery and Development Platform, ADME of Therapeutics facility, Department of Phamracy, Uppsala University, Uppsala, Sweden.
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Lindqvist
Uppsala Drug Optimisation and Pharmaceutical Profiling Platform (UDOPP), Department of Pharmacy, Uppsala University, Uppsala, Sweden.
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Filppula
Institute of biomedical sciences, Academia Sinica, Taipei-115, Taiwan.
T-C
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Lee
Department of Pharmacy and.|Science for Life Laboratory Drug Discovery and Development Platform, ADME of Therapeutics facility, Department of Phamracy, Uppsala University, Uppsala, Sweden.|Uppsala Drug Optimisation and Pharmaceutical Profiling Platform (UDOPP), Department of Pharmacy, Uppsala University, Uppsala, Sweden.
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Artursson
Sahlgrenska Translational Melanoma Group (SATMEG), Sahlgrenska Cancer Center, Department of Surgery, Institute of Clinical Sciences, University of Gothenburg and Sahlgrenska University Hospital, Gothenburg.
J A
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Nilsson
Biomedical Research Foundation of the Academy of Athens, Athens, Greece.|Faculty Institute for Cancer Sciences, Manchester Academic Health Sciences Centre, University of Manchester, Manchester, UK.
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VG
Gorgoulis
Clinical Proteomics Mass Spectrometry, Department of Oncology-Pathology.
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J
Lehtiö
Division of Physiological Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
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Zubarev
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics.
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Scobie
Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics thomas.helleday@scilifelab.se.
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Helleday
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D045744:Cell Line, Tumor; D004247:DNA / Q000235:genetics / Q000378:metabolism; D045643:DNA Repair Enzymes / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000302:isolation & purification / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000627:therapeutic use*; D006801:Humans; D051379:Mice; D009369:Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D009711:Nucleotides / Q000378:metabolism; D010084:Oxidation-Reduction; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics; D011743:Pyrimidines / Q000008:administration & dosage*; D034741:RNA, Small Interfering / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
D004791:Enzyme Inhibitors; D009711:Nucleotides; D011743:Pyrimidines; D034741:RNA, Small Interfering; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf; D010744:Phosphoric Monoester Hydrolases; C078207:8-oxodGTPase; D045643:DNA Repair Enzymes; D003849:Deoxyguanosine
DNA damage; MTH1; cancer; reactive oxygen species; small molecule inhibitors