International journal of molecular sciences, 2020-11, Vol.21 (22), p.8860
2020
Details
- Autor(en) / Beteiligte
- Titel
- Radiolabeled 6-(2, 3-Dichlorophenyl)-N4-methylpyrimidine-2, 4-diamine (TH287): A Potential Radiotracer for Measuring and Imaging MTH1
- Ist Teil von
- International journal of molecular sciences, 2020-11, Vol.21 (22), p.8860
- Ort / Verlag
- Switzerland: MDPI AG
- Erscheinungsjahr
- 2020
- Link zum Volltext
- Quelle
- Free E-Journal (出版社公開部分のみ）
- Beschreibungen/Notizen
- MTH1 (MutT homolog 1) or NUDT1 (Nudix Hydrolase 1), also known as oxidized purine nucleoside triphosphatase, has potential as a biomarker for monitoring cancer progression and quantifying target engagement for relevant therapies. In this study, we validate one MTH1 inhibitor TH287 as a PET MTH1 radiotracer. TH287 was radiolabeled with tritium and the binding of [ H]TH287 to MTH1 was evaluated in live glioblastoma cells (U251MG) through saturation and competitive binding assays, together with in vitro enzymatic assays. Furthermore, TH287 was radiolabeled with carbon-11 for in vivo microPET studies. Saturation binding assays show that [ H]TH287 has a dissociation constant ( ) of 1.97 ± 0.18 nM, of 2676 ± 122 fmol/mg protein for U251MG cells, and of 0.98 ± 0.02. Competitive binding assays show that TH287 ( : 3.04 ± 0.14 nM) has a higher affinity for MTH1 in U251MG cells compared to another well studied MTH1 inhibitor: (S)-crizotinib ( : 153.90 ± 20.48 nM). In vitro enzymatic assays show that TH287 has an of 2.2 nM in inhibiting MTH1 hydrolase activity and a of 1.3 nM from kinetics assays, these results are consistent with our radioligand binding assays. Furthermore, MicroPET imaging shows that [ C]TH287 gets into the brain with rapid clearance from the brain, kidney, and heart. The results presented here indicate that radiolabeled TH287 has favorable properties to be a useful tool for measuring MTH1 in vitro and for further evaluation for in vivo PET imaging MTH1 of brain tumors and other central nervous system disorders.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1422-0067, 1661-6596eISSN: 1422-0067DOI: 10.3390/ijms21228860Titel-ID: cdi_doaj_primary_oai_doaj_org_article_354da1a554fd4da787ad6f0b3eea8901
- Format
- –
- Schlagworte
- 2,4-D, Animals, Assaying, Binding, binding assay, Biomarkers, Biomarkers, Tumor - isolation & purification, Biomarkers, Tumor - metabolism, Brain - diagnostic imaging, Brain - metabolism, Brain cancer, Brain tumors, Cell Line, Tumor, Central nervous system, Competition, Crizotinib - pharmacology, Deoxyribonucleic acid, Diamines, DNA, DNA damage, DNA Repair Enzymes - antagonists & inhibitors, DNA Repair Enzymes - genetics, DNA Repair Enzymes - isolation & purification, Enzymes, Evaluation, Glioblastoma, Glioblastoma - diagnostic imaging, Glioblastoma - genetics, Glioblastoma - pathology, Glioblastoma cells, Heart - diagnostic imaging, Homology, Humans, Hydrolase, In vivo methods and tests, Inhibitors, Kidney - diagnostic imaging, Kidney - metabolism, Ligands, Liver cancer, Magnetic resonance imaging, Medical imaging, Mice, MTH1, Neuroimaging, Oxidative stress, PET, Phosphoric Monoester Hydrolases - antagonists & inhibitors, Phosphoric Monoester Hydrolases - genetics, Phosphoric Monoester Hydrolases - isolation & purification, Positron emission, Positron emission tomography, Positron Emission Tomography Computed Tomography, Purine nucleoside triphosphatase, Pyrimidines - chemistry, Pyrimidines - pharmacology, Radioactive tracers, radiotracer, Saturation, TH287, Tomography, Triphosphatase, Tritium, Tumors